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ESTUDOS
AVANÇADOS
INTERDISCIPLINARES
VOLUME 21
Organizadores:
Robson Antonio Tavares Costa
Estélio Silva Barbosa
ESTUDOS
AVANÇADOS
INTERDISCIPLINARES
VOLUME 21
Organizadores
Organizadores:
Organizador
Dr.Robson
ROBSONAntonio
ANTONIO
TAVARES
COSTA
Tavares
Costa
Robson Antonio Tavares Costa
Estélio Silva Barbosa
EDITORA ENTERPRISING
Direção Nadiane Coutinho
Gestão de Editoração Antonio Rangel Neto
Gestão de Sistemas João Rangel Costa
Conselho Editorial ·Antonio Augusto Teixeira Da Costa, Phd – Ulht – Pt
·Eraldo Pereira Madeiro, Dr – Unitins – Br
·Eugenia Maria Mariano da Rocha Barichello, Dra.
UFSM;
·Luama Socio, Dra. - Unitins - Br
·Ismael Fenner, Dr. - Fics – Py
·Francisco Horácio da Silva Frota, Dr. UECE;
·Tânia Regina Martins Machado, Dra. - Unitins – Br;
·Agnaldo de Sousa Barbosa, Dr. UNESP.
Copyright © 2023 da edição brasileira.
by Editora Enterprising.
Copyright © 2023 do texto.
by Autores.
Todos os direitos reservados.
Todo o conteúdo apresentado neste livro,
inclusive correção ortográfica e gramatical, é
de responsabilidade do(s) autor(es). Obra sob
o selo Creative Commons-Atribuição 4.0
Internacional. Esta licença permite que outros
remixem, adaptem e criem a partir do
trabalho, para fins não comerciais, desde que
lhe atribuam o devido crédito e que licenciem
as novas criações sob termos idênticos.
Diagramação
João Rangel Costa
Design da capa
Nadiane Coutinho
Revisão de texto
Os autores
EDITORA ENTERPRISING
www.editoraenterprising.net
E-mail: contacto@editoraenterprising.net
Tel. : +55 61 98229-0750
CNPJ: 40.035.746/0001-55
Robson Antonio Tavares Costa
Estélio Silva Barbosa
(Organizadores)
Estudos Avançados
Interdisciplinares
Volume 21
Brasília - DF
E82
Estudos Avançados Interdisciplinares Volume 21 / Robson Antonio
Tavares Costa (Organizador), Estélio Silva Barbosa (Organizador)Brasília: Editora Enterprising, 2023.
(Estudos Avançado Interdisciplinares Volume 21)
Livro em PDF
170p., il.
ISBN: 978-65-84546-43-1
DOI: 10.29327/5242655
1.Interdisciplinares. 2. Pesquisas. 3. Práticas. 4. Estudos.
I. Título.
CDD: 370
Acreditamos que o conhecimento é a grande estratégia de inclusão e integração, e a escrita é a grande
ferramenta do conhecimento, pois ela não apenas permanece, ela floresce e frutifica.
Equipe Editora Enterprising.
Sumário
08
APRESENTAÇÃO
CAPÍTULO 1:
A INCLUSÃO DOS ALUNOS COM DEFICIÊNCIA: DESAFIOS
SOCIOPOLÍTICOS E ORIENTAÇÕES NORMATIVO-PEDAGÓGICAS EM
ÂMBITO NACIONAL
09
Railene dos Santos Monteiro
Ricardo Figueredo Pinto
CAPÍTULO 2:
FATORES E MOTIVAÇÕES NA DÁDIVA DE SANGUE NO CONTEXTO
LUSOBRASILEIRO
26
Ivonete Ferreira Maciel
Danilele Ramos Guedes
Simone do Socorro Azevedo Lima
CAPÍTULO 3:
CORRIDA DE RUA E SISTEMA IMUNOLÓGICO RUNNING AND THE
IMMUNE SYSTEM
55
Gabriel Lemos da Costa
Moisés Simão Santa Rosa de Sousa
Virvalene Costa Melo
Ricardo Figueredo Pinto
CAPÍTULO 4:
PERFIL NUTRICIONAL DE PACIENTES RENAIS EM HEMODIÁLISE: UMA
REVISÃO INTEGRATIVA DA LITERATURA
65
Carliane de Jesus Louzeiro
Joseana Moreira Assis Ribeiro
Fábio Costa de Vasconcelos
CAPÍTULO 5:
CONTRIBUIÇÃO DAS ATIVIDADES FÍSICAS SOBRE OS IMPACTOS DO
TRABALHO PROLONGADO EM POSIÇÃO ORTOSTÁTICA EM
TRABALHADORES DO CENTRO COMERCIAL DE BELÉM-PA
Moisés Simão Santa Rosa de Sousa
Natália de Nazaré Martins Vasconcelos
Virvalene Costa Melo
Ricardo Figueredo Pinto
73
CAPÍTULO 6:
IMPORTÂNCIA DA NUTRIÇÃO PARA OS PRATICANTES DE
MUSCULAÇÃO
110
Lucas Levy Silva de Oliveira
Caroline Laila Segundo S. Nascimento
Joseana Moreira Assis Ribeiro
CAPÍTULO 7:
A IMPORTÂNCIA DA AJUDA DOS PAIS NO TRATAMENTO
ODONTOLÓGICO DOS FILHOS
122
Maria Betânia Furtado Araújo
Bruno de Souza Carvalho Tavares
CAPÍTULO 8:
LUTO E A PSICOLOGIA HOSPITALAR
132
Elizandra Vieira Nogueira
Bruno de Souza Carvalho Tavares
CAPÍTULO 9:
CRICOTIREOIDOSTOMIA:
EMERGÊNCIA
CIRÚRGICA
E
POR
PUNÇÃO
NA
144
Laisa Katrine Lemos do Carmo
Bruno de Souza Carvalho Tavares
CAPÍTULO 10:
A IMPORTÂNCIA DA AUDITORIA INTERNA PARA AS ORGANIZAÇÕES
Tayane Carmo da Silva
155
Apresentação
Prezados(as) leitores(as),
É com muita satisfação que apresentamos o vigésimo primeiro
volume
da
Coleção
intitulada
“ESTUDOS
AVANÇADOS
INTERDISCIPLINARES”, que reúne em seus capítulos pesquisadores
de diversas instituições com discussões e temáticas que circundam
uma gama de possibilidades de pesquisas e de relações dialógicas
que certamente podem ser relevantes para o desenvolvimento
social brasileiro a partir de uma ótica que contempla as mais
vastas questões da sociedade. Tal obra visa dar publicidade a
estudos e pesquisas frutos de árduos trabalhos acadêmicos que
decerto contribuem, cada um a seu modo, para o aprofundamento
de discussões em suas respectivas áreas pois são pesquisas
germinadas, frutificadas e colhidas de temas atuais que estão
sendo debatidos nas principais universidades nacionais e que
refletem o interesse de pesquisadores no desenvolvimento social e
científico que possam impactar positivamente a qualidade de vida
de homens e de mulheres.
Assim sendo, convidamos todos os leitores para exercitar diálogos
com os estudos aqui contemplados, esperamos que os textos
publicados contribuam para a formação intelectual e a reflexão
crítica dos alunos, professores e demais leitores. Desejamos
ressaltar, em nome de todos que compõem a Editora Enterprising,
a nossa gratidão para com os pesquisadores cujos trabalhos
aparecem aqui reunidos, que diante da dedicação, temos a
oportunidade de nos debruçar acerca de assuntos atuais e
pertinentes.
Sejam bem-vindos e tenham proveitosas leituras!
Equipe Editora Enterprising.
Capítulo 1
A INCLUSÃO DOS ALUNOS
COM DEFICIÊNCIA: DESAFIOS
SOCIOPOLÍTICOS E
ORIENTAÇÕES NORMATIVOPEDAGÓGICAS EM ÂMBITO
NACIONAL
DOI: 10.29327/5242655.1-1
Railene dos Santos Monteiro
Ricardo Figueredo Pinto
ESTUDOS AVANÇADOS INTERDISCIPLINARES Vol. 21
A INCLUSÃO DOS ALUNOS COM DEFICIÊNCIA: DESAFIOS
SOCIOPOLÍTICOS E ORIENTAÇÕES NORMATIVO-PEDAGÓGICAS EM
ÂMBITO NACIONAL
Railene dos Santos Monteiro
Ricardo Figueredo Pinto
RESUMO
Este artigo tem como objetivo analisar o desafio histórico-social da educação dos alunos com
deficiência na perspectiva da inclusão como direito humano/social, apresentar alguns elementos
fundamentais da política nacional de inclusão no ensino regular e verificar se são adequadas ao
desenvolvimento da capacidade pessoal e social, integral e plena do aluno, conforme reza o artigo
4º, III da LBD/96. Assim utilizou-se como metodologia uma pesquisa documental e bibliográfica a
fim de que houvesse resultados mais concisos e coerentes. De modo, que resultou nas discussões
sobre pressupostos sociopolíticos e orientações normativo-pedagógicas que a fundamentam,
estruturam e organizam a inclusão educacional no âmbito nacional, trazendo em um único estudo
apontamentos sobre a Constituição Federal de 1988; Declaração Mundial de Educação para Todos
de 1990; Declaração de Salamanca de 1994; Lei de Diretrizes e Bases da Educação Nacional de
1996; Resolução CNE/CEB nº 2, de 2001, paralelo ao que já foi produzido sobre a temática no
Brasil. Logo, este artigo trás discussões importantes ao campo da educação inclusiva e abre
horizontes para novos estudos.
Palavras-chaves: Alunos com deficiência. Desafios Sociopolíticos. Inclusão educacional.
ABSTRACT
This article aims to analyze the historical-social challenge of educating students with disabilities
from the perspective of inclusion as a human/social right, to present some fundamental elements of
the national policy of inclusion in regular education and to verify whether they are adequate for the
development of capacity personal and social, integral and full of the student, as stated in article 4,
III of LBD/96. Thus, a documentary and bibliographical research was used as a methodology in
order to have more concise and coherent results. Thus, it resulted in discussions on sociopolitical
assumptions and normative-pedagogical guidelines that underlie, structure and organize educational
inclusion at the national level, bringing in a single study notes on the Federal Constitution of 1988;
World Declaration on Education for All 1990; 1994 Salamanca Declaration; Law of Guidelines and
Bases of National Education of 1996; CNE/CEB Resolution nº 2, from 2001, parallel to what has
already been produced on the subject in Brazil. Therefore, this article brings important discussions
to the field of inclusive education and opens horizons for new studies.
Keywords: Students with disabilities. Sociopolitical Challenges. educational inclusion
10
ESTUDOS AVANÇADOS INTERDISCIPLINARES Vol. 21
RESUMEN
Este artículo tiene como objetivo analizar el desafío histórico-social de educar a los estudiantes con
discapacidad desde la perspectiva de la inclusión como derecho humano/social, presentar algunos
elementos fundamentales de la política nacional de inclusión en la educación regular y verificar si
son adecuados. para el desarrollo de la capacidad personal y social, integral y plena del alumno,
conforme lo dispuesto en el artículo 4, III de la LBD/96. Así, se utilizó como metodología una
investigación documental y bibliográfica para tener resultados más concisos y coherentes. Así,
resultó en discusiones sobre supuestos sociopolíticos y lineamientos normativo-pedagógicos que
sustentan, estructuran y organizan la inclusión educativa a nivel nacional, trayendo en un solo
apuntes de estudio sobre la Constitución Federal de 1988; Declaración Mundial sobre Educación
para Todos 1990; Declaración de Salamanca de 1994; Ley de Lineamientos y Bases de la
Educación Nacional de 1996; Resolución CNE/CEB nº 2, de 2001, paralela a lo ya producido sobre
el tema en Brasil. Por lo tanto, este artículo trae discusiones importantes para el campo de la
educación inclusiva y abre horizontes para nuevos estudios.
Palabras Clave: Estudiantes con discapacidad. Desafíos sociopolíticos. inclusión educative.
1. INTRODUÇÃO
A constituição de uma política nacional de educação inclusiva de pessoas com deficiência
possui um itinerário marcado por desafios sociopolíticos, no sentido de assegurá-la como direito
humano e social, assim como também desafiador é o estabelecimento de diretrizes-paradigmas de
orientações normativo-pedagógicas do processo de ensino-aprendizagem adequados e eficazes a
essa condição pessoal e social, dada a complexidade do fenômeno em questão.
Na concepção tradicional de atendimento pedagógico sustentava-se a integração escolar do
aluno às estruturas físicas, administrativa, curricular, pedagógica e política da escola, onde teria que
ser capaz de aprender no nível pré-estabelecido pelo sistema de ensino.
“Numa perspectiva de integração, o indivíduo com deficiência não é recusado no ambiente
escolar. Ele pode participar, desde que se adapte, desde que reúna condições individuais
necessárias para estar em um dado ambiente.” (CAMARGO, 2008, p. 76-77).
Da integração dessas pessoas na escola, partiu-se para uma concepção de inclusão escolar,
cujo processo de ensino-aprendizagem envolve uma inserção total e incondicional do aluno e exige
a transformação da escola, pois defende a inclusão de alunos com quaisquer deficiências e
necessidades, preferencialmente no sistema de ensino regular, cabendo às escolas adaptarem-se às
suas especificidades.
As escolas inclusivas propõem um modo de organização do sistema educacional que
considera as necessidades de todos os alunos e que é estruturado em função dessas
necessidades. (MANTOAN, 2003, p. 24).
11
ESTUDOS AVANÇADOS INTERDISCIPLINARES Vol. 21
O Brasil fez opção pela construção de um sistema educacional inclusivo ao concordar com a
Declaração Mundial de Educação para Todos, firmada em Jomtien, na Tailândia, em 1990, e ao
mostrar consonância com os postulados produzidos em Salamanca (Espanha, 1994) na Conferência
Mundial sobre Necessidades Educacionais Especiais: Acesso e Qualidade. Desses documentos,
ressaltam-se alguns princípios inspiradores que orientam a política nacional de atendimento de
alunos com deficiência, presentes na Constituição Federal de 1988 (CF/88), na Lei de Diretrizes e
Bases da Educação (1996) e na Resolução CNE/CEB nº 2, de 11 de setembro de 2001, que
estabelece as Diretrizes Nacionais para a Educação Especial na Educação Básica. Deste modo, a
necessidade e o amplo debate sobre a constituição de uma política de educação inclusiva levou o
Brasil a adotar a concepção e proposta de inclusão de alunos com deficiências preferencialmente na
rede regular, conforme preceitua os artigos 208, III da CF/88 e 4º, III e 58 da LDB/96.
O amplo contexto da reflexão histórico-social e teórica, os grandes embates na elaboração e
implantação de paradigmas para as orientações normativo-pedagógicas e a complexidade do
processo de ensino-aprendizagem adequados à essa condição pessoal e social, faz entender que a
inclusão escolar desses alunos é um processo desafiador que envolve todos os atores que atuam e
influem o ambiente institucional escolar (família, pais, amigos, direção, coordenação pedagógica,
serviço especializado, alunos, professores), como também, a interação e inserção nos grupos
sociais (BRASIL/MEC, 2001, p. 6).
O objetivo desse sucinto trabalho é analisar esse desafio da inclusão escolar da pessoa com
deficiência no sentido de assegurar a educação como direito humano e social, como também estudar
de onde emanam e como se dá princípios e as orientações normativo-pedagógicas do processo de
ensino-aprendizagem, tendo como finalidade verificar se são adequadas ao desenvolvimento da
capacidade pessoal e social, integral e plena do aluno, conforme reza o artigo 4º, III e V da LBD/96.
2. O DESAFIO DA EDUCAÇÃO DA PESSOA COM DEFICIÊNCIA COMO DIREITO
HUMANO E SOCIAL
Segundo Sassaki (1997), o processo de inclusão e em particular, o de inclusão educacional,
desencadeou-se a partir da reflexão histórico-social e cultural de conceitos, mentalidades e
comportamentos considerados deficitários/diferentes/minoritários no padrão social. Quatro períodos
norteariam a história das pessoas com deficiência:
12
ESTUDOS AVANÇADOS INTERDISCIPLINARES Vol. 21
a) Período da exclusão: é aquele identificado antes do século XX, onde as pessoas com deficiências
não participavam de qualquer tipo de educação em escolas, sendo pessoas rejeitadas, perseguidas e
ignoradas pela sociedade;
b) Período da segregação: ocorre já no século XX e se estende até mais ou menos à década de
cinquenta. O atendimento às crianças com deficiências é dado exclusivamente nas grandes
instituições especializadas. Na década de 60, surgem as escolas especiais e mais tarde as classes
especiais dentro das escolas comuns. O que faz surgir no sistema educacional brasileiro dois
subsistemas: a educação comum e a educação especial;
c) Período da integração: localizado na década de 70, onde as escolas comuns passaram a aceitar os
alunos com deficiências em salas comuns. Os alunos tinham que se adaptar às escolas, seus
currículos, sua estrutura, enfim, tudo o que ela proporcionava. E aqueles que não se adaptavam
eram então excluídos. Surgem as classes especiais dentro das escolas regulares;
d) Período da Inclusão: surge na forma em que se encontra hoje; tem seu início a partir da segunda
metade da década de 80, implementou-se na década de 90 e permeia as ideias deste século (XXI) e
abre possibilidades acerca da visão futura. Neste último período, a proposta é de adaptar a escola
para receber os alunos com deficiência, onde as diferenças devem ser aceitas e, assim, a escola
comum se tornar inclusiva.
Tratando-se de Brasil, vê-se que sua história da educação está fortemente marcada pela
exclusão escolar. Desde a Colonização, os alunos eram diferenciados e classificados de acordo com
a sua classe social, cor, gênero, raça, tipo de deficiência entre outras classificações excludentes. As
oportunidades eram para poucos e somente a elite tinha acesso à escola, sendo este o quadro
educacional do país até a primeira metade do século XX (MAZZOTA, 2005, p. 16).
“Falar sobre a educação especial no Brasil implica, necessariamente, a consideração de dois
aspectos constitutivos de nossa historia: a desigualdade e a diversidade. O país é construído
a partir da diversidade de populações e de suas histórias, mas de forma extremamente
desigual. A formação econômica do Brasil e as características de sua organização social
fizeram com que o país passasse a conviver com vários problemas que impactaram
diretamente a vida de crianças e jovens brasileiros, muitos dos quais presentes ate hoje:
crianças abandonadas nas ruas das grandes cidades, desde o século XVII (Lima &
Venâncio, 1991), restrita cobertura escolar e, consequentemente, um numero grande de
analfabetos (Gomes, 2001; Lima, 2011), entre outros. Findado o regime escravocrata,
muitas famílias não foram incorporadas diretamente ao setor produtivo, passando a
sobreviver nas grandes cidades, sem acesso a condições de vida minimamente satisfatórias.
Na República, a massa de brasileiros desempregada e considerada iletrada foi identificada
como marginal e seus hábitos vistos como indecentes e de transgressão aos bons costumes,
13
ESTUDOS AVANÇADOS INTERDISCIPLINARES Vol. 21
aos olhos de uma elite que tomava seu país como atrasado em relação a Europa.”
Somente a partir da segunda metade do século XX, com a expansão de instituições privadas
de educação especial (de cunho assistencialista) (ROSADO, 2009, p. 3), as mudanças sóciopolíticas da década de 1950/60 (período pós-guerra “era de ouro”, desenvolvimentismo moderno de
JK) e a grande demanda de “pessoas especiais” que não conseguiam ser incluídas nas instituições
públicas, começou-se a refletir no meio político e a elaborar, de forma organizada e
institucionalizada, o direito à educação escolar das “pessoas com deficiência”, mas com o objetivo
de adequar o sistema educacional nacional ao discurso da formação de recursos humanos para o
desenvolvimento do país, numa referência à teoria do capital humano (PUZIOL e SILVA, 2013):
“Em 1967, a primeira Constituição após o Golpe de Estado de 1964 previu o
estabelecimento dos planos nacionais de educação. A Emenda Constitucional de 1969
estabeleceu a execução dos planos nacionais e regionais de desenvolvimento. A legislação
sobre as diretrizes e bases da educação nacional foi revista e, em 1971, a Lei Educacional
n° 5.692 passou a obrigatoriedade da escolarização brasileira para oito anos. Esse período
foi decisivo para o início da formatação da Educação Especial como uma política de
Estado, com a criação de um órgão vinculado ao Ministério da Educação e Cultura, o
Centro Nacional de Educação Especial (CENESP), que passou a ter a responsabilidade de
formular e impulsionar as ações de Educação Especial no Brasil. A criação deste órgão e a
implantação de suas ações encontraram subsídio na perspectiva desenvolvimentista adotada
pelo regime militar à época” (KASSAR, 2001, p. 44).
Nesse contexto, por meio de Campanhas Nacionais (ROSADO, 2009, p. 5) e de amplo debate
para promover um atendimento educacional mais adequado aos deficientes em todo o território
nacional, a educação especial começa a se constituir enquanto direito subjetivo e social. A primeira
Lei de Diretrizes e Bases da Educação (Lei nº. 4.024/61), estabelece a diretriz normativa e
pedagógica de incluir preferencialmente o aluno “excepcional” no sistema educacional comum, mas
de ainda contar com as parcerias de instituições privadas, demostrando ainda a omissão do Estado,
como se podem ver no Título X, artigos 88 e 89 da referida lei:
“Art. 88. A educação de excepcionais deve no que for possível, enquadrar-se no sistema
geral de educação, a fim de integrá-los na comunidade.
Art. 89. Toda iniciativa privada considerada eficiente pelos conselhos estaduais de
educação, e relativa à educação de excepcionais, receberá dos poderes públicos tratamento
14
ESTUDOS AVANÇADOS INTERDISCIPLINARES Vol. 21
especial mediante bolsas de estudo, empréstimos e subvenções.”
Em 1971, com a reforma do ensino de 1º e 2º graus, a LDB de 1961 foi alterada, passando a
vigorar a Lei 5.692/71. Nesta Lei, a educação direcionada aos deficientes está presente no artigo 9º,
referente ao capítulo do ensino de 1º e 2º graus, significando que a educação dos “excepcionais”
está incluída na educação básica.
Com o advento da CF/88, é definida a educação como um direito de todos, assegurando o
pleno desenvolvimento da pessoa, o exercício da cidadania e a qualificação para o trabalho (artigo
205). É estabelecida a igualdade de condições de acesso e permanência na escola como um dos
princípios para o ensino (artigo 206, inciso I). A garantia do direito à educação das pessoas com
deficiência é dever do Estado e sua oferta de atendimento educacional especializado deve acontecer
preferencialmente na rede regular de ensino (artigo 208, inciso III). Fica definida, portanto, uma
concepção de educação inclusiva numa perspectiva ampla com atendimento a todas as pessoas em
idade escolar, tendo o Estado (União, Estados, Municípios e Distrito Federal) como responsável por
garantir o direito à educação e prestar o serviço às pessoas com deficiências preferencialmente no
ensino regular.
Convém lembrar que a proposta de uma política nacional de educação de pessoas com
deficiências como direito humano e social, conforme os referidos diplomas e dispositivos legais,
segundo Laplane (2004, p. 18), ainda está distante de ser alcançado efetivamente, devido a um
conjunto de fatores que delineiam um quadro complexo de exclusão social e econômico no país,
que levam, muitas vezes, o atendimento aos moldes tradicionais de não-segregação (integração ao
sistema educacional pré-estabelecido) ou a verdadeira ausência de atendimento educacional:
O discurso contradiz a realidade educacional brasileira, caracterizada por salas
superlotadas, instalações físicas insuficientes, quadros docentes cuja formação deixa a
desejar. Essas condições de existência do nosso sistema educacional levam a questionar a
própria ideia de inclusão como política que, simplesmente, insira alunos nos contextos
escolares existentes.
Sobre esse quadro complexo de exclusão social e econômico no país, Kassar (2012, p. 11) se
expressa:
“Na última década, diferentes trabalhos (Goncalves, 2008; Kassar, 2006; Pletsch, 2010)
apontam para situações de fracasso de alunos com deficiências nas escolas comuns,
inclusive quando todos os quesitos previstos pela legislação educacional estão presentes
(professores formados, salas adaptadas, número de alunos reduzidos por sala, frequência
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ESTUDOS AVANÇADOS INTERDISCIPLINARES Vol. 21
em salas de recursos multifuncionais no contraturno escolar, entre outros). Os resultados
encontrados em pesquisas brasileiras parecem discrepar dos resultados de pesquisas em
outros países de economia central, como as relatadas por Downing e MacFarland (2010).
No entanto, uma análise que enfoque a escolaridade das crianças com deficiências em
contraste com a educação geral brasileira nos permite dizer que aqueles alunos não são os
únicos a não demonstrar bom desempenho escolar. Expandindo nosso foco para o ensino
fundamental brasileiro, e possível identificar vários problemas. Inicialmente, pode-se
apontar sua ineficiência em relação a matricular – de fato – toda a população. Lima (2011)
explica que, desde a década de 1990, apesar do ensino fundamental brasileiro apresentar
condições físicas (capacidade instalada) para atender a todos os indivíduos na faixa etária
adequada (numero de escolas sufi cientes), dado o considerável numero de indivíduos fora
da faixa etária na escola (distorção idade x série) e de indivíduos em idade própria fora
dela, ainda não cumpriu esta obrigação. As análises de Pinto e Alves (2010) também
contribuem para entender a dinâmica presente em nossas escolas. Além dos problemas
apontados por Lima (2011), temos baixo investimento de recursos em educação pública,
quando comparamos recursos do Fundo de Manutenção e Desenvolvimento da Educação
Básica e de Valorização dos Profissionais da Educação (Fundeb) de diferentes estados e o
investimento em educação pública de países de economia central e mesmo de escolas de
elite do estado de São Paulo.”
Apesar das políticas públicas preconizarem a garantia de acesso escolar e a aceitação da
pessoa com deficiência, com um esforço coletivo na equiparação de oportunidades, a literatura
aponta muitas dificuldades para sua implementação na realidade brasileira. Esse descompasso entre
o que está na legislação e o que se constata na prática leva a pensar o desafio da inclusão escolar
como parte da inclusão social e econômica, sendo um “processo pelo qual a sociedade se adapta
para poder incluir, em seus sistemas sociais gerais, pessoas com necessidades especiais e,
simultaneamente, estas se preparam para assumir seus papéis na sociedade” (SASSAKI, 1997, p.
41).
Há uma clara necessidade de que os sujeitos da escola e da comunidade participem de sua
construção, reconhecendo as diferenças entre os alunos de modo a desenvolver um projeto político
pedagógico a partir de suas necessidades e das experiências vivenciadas na sala de aula, tendo como
garantia a qualidade da educação básica para uma formação integral e plena de acordo com a
capacidade de cada um (LDB/96, artigo 4º, V).
Portanto, para construir uma educação que abranja todos os segmentos da sociedade e cada
um dos seus cidadãos é necessária uma estratégia de ação que tenha princípios e diretrizes
normativo-pedagógicas acerca do processo de ensino-aprendizagem de alunos com deficiência
baseado na inclusão de todos, quaisquer que sejam suas limitações e possibilidades individuais e
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sociais.
3. PRINCÍPIOS
E
DIRETRIZES
NORMATIVO-PEDAGÓGICAS
ACERCA
DO
PROCESSO DE ENSINO-APRENDIZAGEM DE ALUNOS COM DEFICIÊNCIA
Para uma ampla e profunda análise dos princípios e diretrizes normativo-pedagógicas que
orientam o processo de ensino-aprendizagem em âmbito nacional de pessoas com deficiência, além
do estudo dos dois grandes marcos referenciais nacionais - a Lei de Diretrizes e Bases da Educação
Nacional (LDB/96) e a Resolução CNE/CEB n.º 02, de 11 de setembro de 2001, que institui as
Diretrizes Nacionais para a Educação Especial na Educação Básica -, faz-se necessário estudar
alguns marcos referenciais internacionais, considerados pressupostos sociopolíticos e princípios
inspiradores dessas diretrizes normativas e pedagógicas nacionais.
No Brasil, a apropriação do discurso favorável à inclusão foi fortemente influenciada por
movimentos e declarações internacionais, desde o final da década de 40, com a Declaração
Universal dos Direitos Humanos, tomando maior impulso a partir dos anos 90 em favor da
implantação das reformas neoliberais, com a já citada teoria do capital humano a favor das teses
desenvolvimentistas do país (PUZIOL e SILVA, 2013).
A Assembleia Geral da Organização das Nações Unidas produziu vários documentos
internacionais, norteadores para o desenvolvimento de políticas públicas de seus países membros,
entre esses, o Brasil, que reconhece seus conteúdos e os respeita na elaboração das políticas
públicas nacionais. Dentre os documentos produzidos, cita-se: Declaração Universal dos Direitos
Humanos (1948); Declaração Mundial Sobre Educação para Todos (1990); Declaração de
Salamanca (1994); Convenção Interamericana para a Eliminação de Todas as Formas de
Discriminação Contra as Pessoas Portadoras de Deficiência (Convenção da Guatemala/1999) e a
Declaração de Montreal sobre a Inclusão (Canadá/2001). Destes, apresentar-se-á alguns pontos
relevantes da Declaração Mundial Sobre Educação para Todos e da Declaração de Salamanca.
Em 1990, organismos internacionais (Banco Mundial, Programa das Nações Unidas para o
Desenvolvimento - PNUD, Fundo das Nações Unidas para a Infância - UNICEF e Organização das
Nações Unidas para a Educação e Cultura - UNESCO) organizaram a Conferência de Educação
Mundial Para Todos (Conferência de Jomtien/Tailândia), em que foram discutidos problemas
relativos à escolaridade da população nos países em desenvolvimento. Ao seu término, foi aprovada
a Declaração de Jomtien (com 10 artigos e o Plano de Ação com 50 itens) onde foram propostas
metas a serem cumpridas pelos países participantes e signatários. Essa Conferência destacou, em
especial, o princípio da universalização e da equidade de acesso à educação, bem expresso no seu
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artigo 3º – Universalizar o acesso à educação e promover a equidade (ONU/UNICEF, 1990):
“1. A educação básica deve ser proporcionada a todas as crianças, jovens e adultos. Para
tanto, é necessário universalizá-la e melhorar sua qualidade, bem como tomar medidas
efetivas para reduzir as desigualdades. 2. Para que a educação básica se torne equitativa, é
mister oferecer a todas as crianças, jovens e adultos, a oportunidade de alcançar e manter
um padrão mínimo de qualidade da aprendizagem. 3. A prioridade mais urgente é melhorar
a qualidade e garantir o acesso à educação para meninas e mulheres, e superar todos os
obstáculos que impedem sua participação ativa no processo educativo. Os preconceitos e
estereótipos de qualquer natureza devem ser eliminados da educação. 4. Um compromisso
efetivo para superar as disparidades educacionais deve ser assumido. Os grupos excluídos –
os pobres; os meninos e meninas de rua ou trabalhadores; as populações das periferias
urbanas e zonas rurais; os nômades e os trabalhadores migrantes; os povos indígenas; as
minorias étnicas, raciais e linguísticas; os refugiados; os deslocados pela guerra; e os povos
submetidos a um regime de ocupação – não devem sofrer qualquer tipo de discriminação no
acesso às oportunidades educacionais. 5. As necessidades básicas de aprendizagem das
pessoas portadoras de deficiências requerem atenção especial. Posteriormente, em 1994, a Conferência Mundial Sobre Necessidades Educativas Especiais:
Acesso e Qualidade, ocorrida em Salamanca/Espanha, sistematizada em 85 artigos em três grandes
partes, teve como resultado a Declaração de Salamanca Sobre Princípios, Política e Práticas na Área
das Necessidades Educativas Especiais. O documento aprovado amplia o conceito de necessidades
especiais na perspectiva da inclusão, inserindo crianças excluídas da escola por trabalho infantil e
abuso sexual e as que têm necessidades especiais graves, afirmando que todas devem ser atendidas
no mesmo ambiente de ensino. Sua Estrutura de Ação em Educação Especial é orientada por vários
princípios, entre esse o de que as escolas devem “acolher todas as crianças, independente de suas
condições físicas, intelectuais, sociais, emocionais, linguísticas ou outras” (artigo 3º).
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Foi durante esta Conferência que o conceito de escola inclusiva passou a ser discutido de
forma mais sistemática, de uma educação acolhedora, adaptada ao aluno com deficiência, bem
articulada politicamente e preparada para receber e ensinar todos em suas singularidades e
particularidades, sendo a escola que deve se adaptar ao aluno e não o contrário, como
historicamente tem acontecido, estabelecendo como princípio fundamental da escola inclusiva
(artigo 7º):
“todas as crianças devem aprender juntas, sempre que possível, independentemente de
quaisquer dificuldades ou diferenças que elas possam ter. Escolas inclusivas devem
reconhecer e responder às necessidades diversas de seus alunos, acomodando ambos os
estilos e ritmos de aprendizagem e assegurando uma educação de qualidade a todos através
de um currículo apropriado, arranjos organizacionais, estratégias de ensino, uso de recurso
e parceria com as comunidades. Na verdade, deveria existir uma continuidade de serviços e
apoio proporcional ao contínuo de necessidades especiais encontradas dentro da escola”
(UNESCO, 1994).
Nos artigos 8º e 9º a Declaração de Salamanca traça orientações referentes à escola especial
e a escola inclusiva:
“8. O encaminhamento de crianças a escolas especiais ou a classes especiais ou a sessões
especiais dentro da escola em caráter permanente deveriam constituir exceções, a ser
recomendado somente naqueles casos infrequentes onde fique claramente demonstrado que
a educação na classe regular seja incapaz de atender às necessidades educacionais ou
sociais da criança ou quando sejam requisitados em nome do bem-estar da criança ou de
outras crianças. 9. Finalmente, escolas especiais ou unidades dentro das escolas inclusivas
podem continuar a prover a educação mais adequada a um número relativamente pequeno
de crianças portadoras de deficiências que não possam ser adequadamente atendidas em
classes ou escolas regulares” (UNESCO, 1994).
Com base nos artigos 3º, 7º, 8º e 9º, percebe-se a concepção de inclusão desta Declaração que
reconhece o princípio de igualdade de oportunidade para todas as pessoas com necessidas especiais
ou não, sempre que possível em escolas regulares e que, exceções a essa regra, devem ser
consideradas individualmente, caso a caso, quando a educação em instituição específica seja
requerida. E ainda que, nos casos excepcionais das escolas especiais, a educação não precisa ser
inteiramente segregada, estabelecendo um fluxo de movimento da escola especial para a regular e a
organização de um trabalho integrado.
Nesse contexto sociopolítico macro, as Declarações de Jomtien e de Salamanca influenciaram
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fortemente a elaboração e a promulgação da LDB/96, em seus treze títulos e cento e vinte artigos,
considerada uma lei baseada no princípio do direito universal à educação. A LDB/96, no Título II,
Dos Princípios e Fins da Educação Nacional, art. 2º e 4º; Titulo III, Do Direito à Educação e do
Dever de Educar, artigos 4º e 5º; e no Título IV, Da Organização da Educação Nacional, artigo 8º,
10º e 11º, fundamenta-se em princípios da CF/88, das Declarações de Jomtien e de Salamanca. O
Estado (União, os Estados, o Distrito Federal e os Municípios) é garantidor de direitos e prestador
de serviços a todos às pessoas e deve se organizar em regime de colaboração entre os seus entes
federados para efetivo atendimento aos mesmos.
A nova LDB/96 reservou um capítulo específico para a educação especial, o Capítulo V Da
Educação Especial, artigos 58, 59 e 60, fato relevante para uma área tão pouco contemplada
historicamente no conjunto das políticas públicas educacionais no Brasil.
Contudo, ao se analisar o Capítulo V desta Lei, percebe-se que o artigo 58 caracteriza a
educação especial como “modalidade especial”, definição que apresenta um “caráter circular, vago
e genérico”, pois prevê, nos parágrafos 1º e 2º, a existência de apoio especializado no ensino regular
e de serviços especiais separados quando não for possível a inclusão.
O artigo 59 aponta as providências ou apoio, de ordem escolar ou de assistência, que os
sistemas de ensino deverão assegurar aos alunos considerados público alvo da educação especial.
Preconiza que os sistemas de ensino devem assegurar aos alunos currículo, métodos, recursos e
organização específica para atender às suas necessidades; assegura a terminalidade específica
àqueles que não atingiram o nível exigido para a conclusão do ensino fundamental, em virtude de
suas deficiências; e garante a aceleração de estudos aos superdotados para conclusão do programa
escolar.
O artigo 60 prevê o estabelecimento de critérios de caracterização das instituições privadas de
educação especial, através dos órgãos normativos dos sistemas de ensino, para o recebimento de
apoio técnico e financeiro público; ao mesmo tempo em que reafirma, em seu parágrafo único, a
preferência pela ampliação do atendimento no ensino regular público.
Além desses artigos que tratam especificamente da educação especial, aponta-se, também,
como fundamentais para o estabelecimento de princípios da inclusão escolar de pessoas com
deficiências, o artigo 3º, inciso I, que garante igualdade de condições para o acesso e permanência
na escola e inciso III, que afirma o direito ao pluralismo de ideias e de concepções pedagógicas; e o
artigo 4º, inciso III, que garante o atendimento educacional especializado gratuito aos educandos
com deficiência, transtornos globais do desenvolvimento e altas habilidades ou superdotação,
transversal a todos os níveis, etapas e modalidades, preferencialmente na rede regular de ensino, e
inciso V, que assegura acesso aos níveis mais elevados do ensino, da pesquisa e da criação artística,
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segundo a capacidade de cada um.
Para implementar e viabilizar esse processo de mudanças sociopolíticas e contribuir para a
normatização da proposta de inclusão educacional prevista na LDB/96, é expedida pela Câmara
Nacional de Educação (CNE), através de sua Câmara de Educação Básica (CEB), a Resolução
CNE/CEB nº 2 de 11 de setembro de 2001, que estabelece as Diretrizes Nacionais para a Educação
Especial na Educação Básica, com base no Parecer CNE/CEB nº 17/2001. A finalidade é atender
alunos que apresentem deficiência em todas as suas etapas e modalidades, tendo início na educação
infantil, nas creches e pré-escolas, assegurando-lhes os serviços de educação especial sempre que se
evidencie, mediante avaliação e interação com a família e a comunidade, a necessidade de
atendimento educacional especializado.
Através desta Resolução, as Diretrizes Nacionais para Educação Especial na Educação Básica
assumem a educação especial como modalidade da educação escolar, entendendo-a como:
“um processo educacional definido por uma proposta pedagógica que assegure recursos e
serviços educacionais especiais, organizados institucionalmente para apoiar, complementar,
suplementar e, em alguns casos, substituir os serviços educacionais comuns, de modo a
garantir a educação escolar e promover o desenvolvimento das potencialidades dos
educandos que apresentam necessidades educacionais especiais, em todas as etapas e
modalidades da educação básica” (BRASIL, 2001, artigo 3º).
Segundo o artigo 4º, a educação especial deverá considerar as situações singulares, os perfis
dos estudantes, as características biopsicossociais dos alunos e suas faixas etárias e se pautar em
princípios éticos, políticos e estéticos de modo a assegurar:
“I - a dignidade humana e a observância do direito de cada aluno de realizar seus projetos
de estudo, de trabalho e de inserção na vida social; II - a busca da identidade própria de
cada educando, o reconhecimento e a valorização das suas diferenças e potencialidades,
bem como de suas necessidades educacionais especiais no processo de ensino e
aprendizagem, como base para a constituição e ampliação de valores, atitudes,
conhecimentos, habilidades e competências; III - o desenvolvimento para o exercício da
cidadania, da capacidade de participação social, política e econômica e sua ampliação,
mediante o cumprimento de seus deveres e o usufruto de seus direitos”.
As especificidades do aluno com público alvo da educação especial seriam aquelas vistas
durante o processo educacional, tais como: I - dificuldades acentuadas de aprendizagem ou
limitações no processo de desenvolvimento que dificultem o acompanhamento das atividades
curriculares, compreendidas em dois grupos: a) aquelas não vinculadas a uma causa orgânica
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específica; b) aquelas relacionadas a condições, disfunções, limitações ou deficiências; II –
dificuldades de comunicação e sinalização diferenciadas dos demais alunos, demandando a
utilização de linguagens e códigos aplicáveis; III - altas habilidades/superdotação, grande facilidade
de aprendizagem que os leve a dominar rapidamente conceitos, procedimentos e atitudes (artigo 5º).
Segundo o referido documento, os fundamentos da proposta para uma política “inclusiva”
devem valorizar a igualdade de oportunidade e a diversidade do processo educativo, determinando
que as escolas da rede regular de ensino devem prever e prover na organização de suas classes
comuns, entre outros pontos, os seguintes (artigo 8º):
“I - professores das classes comuns e da educação especial capacitados e especializados,
respectivamente, para o atendimento às necessidades educacionais dos alunos;
II - distribuição dos alunos com necessidades educacionais especiais pelas várias classes do
ano escolar em que forem classificados, de modo que essas classes comuns se beneficiem
das diferenças e ampliem positivamente as experiências de todos os alunos, dentro do
princípio de educar para a diversidade;
III – flexibilizações e adaptações curriculares que considerem o significado prático e
instrumental dos conteúdos básicos, metodologias de ensino e recursos didáticos
diferenciados e processos de avaliação adequados ao desenvolvimento dos alunos que
apresentam necessidades educacionais especiais, em consonância com o projeto
pedagógico da escola, respeitada a freqüência obrigatória;
VII – sustentabilidade do processo inclusivo, mediante aprendizagem cooperativa em sala
de aula, trabalho de equipe na escola e constituição de redes de apoio, com a participação
da família no processo educativo, bem como de outros agentes e recursos da comunidade.”
Dessa forma, com essas diretrizes nacionais, o que se pretende é garantir que todos os alunos
tenham acesso escolar e aceitação; que as diferenças sejam acolhidas e que ocorra um esforço
coletivo na equiparação de oportunidades de desenvolvimento com qualidade. Não basta que uma
criança público alvo da educação especial seja incluída em uma escola regular para que haja uma
inclusão progressiva e se promova o sucesso escolar do aluno. Para tanto se faz premente que as
condições necessárias para uma educação de qualidade para todos sejam efetivadas, como consta na
Legislação Educacional e nas Diretrizes Nacionais para a Educação Especial na Educação Básica.
4. CONSIDERAÇÕES FINAIS
Conforme analisado neste trabalho, vivencia-se um momento em que mundialmente se debate
e articula a inclusão dos alunos com deficiência preferencialmente na rede regular de ensino.
Constitui-se, na realidade, um desafio histórico-social a educação na perspectiva da inclusão como
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direito humano e social.
A legislação educacional nacional, com seus princípios e diretrizes normativas, são explícitas
quanto à obrigatoriedade em acolher e matricular todos os alunos, independente de suas
necessidades ou diferenças. Entretanto, não é suficiente apenas esse acolhimento, mas que o aluno
tenha condições efetivas de aprendizagem e de desenvolvimento de suas potencialidades. É
necessário e urgente, assim, que os sistemas de ensino se organizem para além de assegurar essas
matrículas, assegurem também a permanência de todos os alunos, sem perder de vista a
intencionalidade pedagógica e a qualidade do ensino, desenvolvendo mecanismos didáticopedagógicos (operacionais) adequados a essa condição pessoal e social.
O princípio da inclusão representa um grande desafio e demanda mudanças em diversos
níveis, desde o diagnóstico dos alunos, a composição e formação de recursos humanos, passando
pelos currículos, pela metodologia de ensino, até a participação da família e da comunidade escolar
na elaboração do projeto político pedagógico.
Como foi discutido quando se tratou da educação como direito humano e social, a inclusão é
um processo histórico-social-político e, como tal, vem para desconstituir paradigmas passados de
segregação e exclusão. A parceria entre ensino comum e ensino especializado é indicada como uma
possibilidade para a melhoria da educação em geral. A inclusão que se pretende efetivar nas escolas
só é possível onde houver respeito às diferenças e, consequentemente, práticas pedagógicas que
permitam a todas as crianças aprenderem de acordo com seu ritmo e suas potencialidades.
Este trabalho trás contribuições significativas e concisas sobre a temática de inclusão
educacional. Para além deste estudo, sugere-se uma análise detalhada de outros documentos:
nacionais como, tais como, resolução nº 4, de 2 de outubro de 2009 (institui diretrizes operacionais
para o atendimento educacional especializado na educação básica, modalidade
de educação
especial ); decreto nº 7.611, de 17 de novembro de 2011( dispõe sobre a educação especial, o
atendimento educacional especializado; lei nº 13.146/2015(institui a lei brasileira de inclusão da
pessoa com deficiência (Estatuto da Pessoa com Deficiência).
REFERÊNCIAS
BEYER, Hugo Otto. A Educação Inclusiva: incompletudes escolares e perspectivas de ação.
Revista de Educação Especial do Centro de Educação, 2012. Disponível em:
<https://periodicos.ufsm.br/educacaoespecial/article/view/5003>. Acesso em: 22/05/2023.
BRASIL. Lei nº. 4.024, de 20 de dezembro de 1961. Fixa as diretrizes e bases da educação
nacional.
Brasília,
DF,
1961.
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em:
<http://www.
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BRASIL. Resolução CNE/CEB nº 2, de 11 de setembro de 2001. Institui Diretrizes Nacionais
para a Educação Especial na Educação Básica. Brasília, DF, 2001. Disponível em:
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Básica.
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em:
<http://portal.mec.gov.br/seesp/arquivos/pdf/parecer17.pdf>. Acesso em: 22/05/2023.
BRASIL. Secretaria de Educação Fundamental. Parâmetros Curriculares Nacionais: Adaptações
Curriculares. Estratégias para a Educação de Alunos com Necessidades Educacionais
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CAMARGO, Éder Pires. Ensino de Física e Deficiência Visual: Dez anos de investigação no
Brasil. São Paulo: Plêiade/FAPESP, 2008.
GIL, A. C. Métodos e técnicas da pesquisa social. 6. ed. São Paulo: Atlas, 2008.
KASSAR, Mônica de Carvalho Magalhães. Educação especial no Brasil: desigualdades e
desafios no reconhecimento da diversidade. Portal SCIELO, São Paulo, 2012. Disponível em:
<http://www.scielo.br/pdf/es/v33n120/10.pdf>. Acesso em: 22/05/2023.
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Development of an Automated Imaging Pipeline for the Analysis of the Zebrafish Larval Kidney
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Jens H. Westhoff
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English
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Spoken
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Jens H. Westhoff1*, Stefan Giselbrecht2, Miriam Schmidts3, Sebastian Schindler1, Philip L. Beales3,
Burkhard Tönshoff1, Urban Liebel4,6*, Jochen Gehrig5,6* Jens H. Westhoff1*, Stefan Giselbrecht2, Miriam Schmidts3, Sebastian Schindler1, Phil
Burkhard Tönshoff1, Urban Liebel4,6*, Jochen Gehrig5,6* 1 Department of Pediatrics I, University Children’s Hospital, University of Heidelberg, Heidelberg, Germany;, 2 Institute for Biological Interfaces , Karlsruhe
Institute of Technology, Eggenstein-Leopoldshafen, Germany, 3 Molecular Medicine Unit, Birth Defects Research Centre, University College London (UCL),
Institute of Child Health, London, United Kingdom, 4 Institute for Applied Computer Science, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen,
Germany, 5 Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, Germany, 6 Accelerator Laboratory, Innovation
Department, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, Germany; Abstract 241955) to PLB, the Stiftung Landesbank Baden-Württemberg to JHW, the Dutch Kidney foundation (CP11.18) to PLB and MS and institutional
funding from Innovation Department, Karlsruhe Institute of Technology (KIT), Germany. MS was funded by an Action Medical Research UK clinical training
fellowship (RTF-1411) and PLB is a Wellcome Trust Senior Research Fellow. We acknowledge support by Deutsche Forschungsgemeinschaft and Open
Access Publishing Fund of Karlsruhe Institute of Technology. The funders had no role in study design, data collection and analysis, decision to publish, or
preparation of the manuscript. Competing interests: The authors have the following conflicts. UL is co-founder and executive director, and JG is application strategist at Acquifer GmbH. There are no patents, products in development or marketed products to declare. This does not alter the authors' adherence to all the PLOS ONE policies
on sharing data and materials. * E-mail: jens.westhoff@med.uni-heidelberg.de (JHW); urban.liebel@kit.edun (UL); jochen.gehrig@kit.edu (JG) Received July 25, 2013; Accepted October 21, 2013; Published December 4, 2013 Copyright: © 2013 Westhoff et al. This is an open-access article distributed under the terms of the Creative Commons Attributi
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. thoff et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
tion, and reproduction in any medium, provided the original author and source are credited. Funding: This project was supported by the European Commission Seventh Framework Programme funded projects DOPAMINET (http://
www.dopaminet.eu/; grant no. 223744) and Eurenomics (http://www.eurenomics.eu/; grant no. 305608) to UL, and SYSCILIA (http://www.syscilia.org/;
grant no. 241955) to PLB, the Stiftung Landesbank Baden-Württemberg to JHW, the Dutch Kidney foundation (CP11.18) to PLB and MS and institutional
funding from Innovation Department, Karlsruhe Institute of Technology (KIT), Germany. MS was funded by an Action Medical Research UK clinical training
fellowship (RTF-1411) and PLB is a Wellcome Trust Senior Research Fellow. We acknowledge support by Deutsche Forschungsgemeinschaft and Open
Access Publishing Fund of Karlsruhe Institute of Technology. The funders had no role in study design, data collection and analysis, decision to publish, or
preparation of the manuscript. Competing interests: The authors have the following conflicts. UL is co-founder and executive director, and JG is application strategist at Acquifer GmbH. There are no patents, products in development or marketed products to declare. Abstract This does not alter the authors' adherence to all the PLOS ONE policies
on sharing data and materials. * E-mail: jens.westhoff@med.uni-heidelberg.de (JHW); urban.liebel@kit.edun (UL); jochen.gehrig@kit.edu (JG) Citation: Westhoff JH, Giselbrecht S, Schmidts M, Schindler S, Beales PL, et al. (2013) Development of an Automated Imaging Pipeline for the Analysis of
the Zebrafish Larval Kidney. PLoS ONE 8(12): e82137. doi:10.1371/journal.pone.0082137 Abstract The analysis of kidney malformation caused by environmental influences during nephrogenesis or by hereditary
nephropathies requires animal models allowing the in vivo observation of developmental processes. The zebrafish
has emerged as a useful model system for the analysis of vertebrate organ development and function, and it is
suitable for the identification of organotoxic or disease-modulating compounds on a larger scale. However, to fully
exploit its potential in high content screening applications, dedicated protocols are required allowing the consistent
visualization of inner organs such as the embryonic kidney. To this end, we developed a high content screening
compatible pipeline for the automated imaging of standardized views of the developing pronephros in zebrafish
larvae. Using a custom designed tool, cavities were generated in agarose coated microtiter plates allowing for
accurate positioning and orientation of zebrafish larvae. This enabled the subsequent automated acquisition of stable
and consistent dorsal views of pronephric kidneys. The established pipeline was applied in a pilot screen for the
analysis of the impact of potentially nephrotoxic drugs on zebrafish pronephros development in the Tg(wt1b:EGFP)
transgenic line in which the developing pronephros is highlighted by GFP expression. The consistent image data that
was acquired allowed for quantification of gross morphological pronephric phenotypes, revealing concentration
dependent effects of several compounds on nephrogenesis. In addition, applicability of the imaging pipeline was
further confirmed in a morpholino based model for cilia-associated human genetic disorders associated with different
intraflagellar transport genes. The developed tools and pipeline can be used to study various aspects in zebrafish
kidney research, and can be readily adapted for the analysis of other organ systems. Citation: Westhoff JH, Giselbrecht S, Schmidts M, Schindler S, Beales PL, et al. (2013) Development of an Automated Imaging Pipeline for the Analysis of
the Zebrafish Larval Kidney. PLoS ONE 8(12): e82137. doi:10.1371/journal.pone.0082137
Editor: Sheng-Ping Lucinda Hwang, Institute of Cellular and Organismic Biology, Taiwan
Received July 25, 2013; Accepted October 21, 2013; Published December 4, 2013
Copyright: © 2013 Westhoff et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This project was supported by the European Commission Seventh Framework Programme funded projects DOPAMINET (http://
www.dopaminet.eu/; grant no. 223744) and Eurenomics (http://www.eurenomics.eu/; grant no. 305608) to UL, and SYSCILIA (http://www.syscilia.org/;
grant no. Automated Screening Platform for Zebrafish Kidneys Despite rodent studies and observational reports in humans,
detailed
data
on
potential
harmful
side
effects
on
nephrogenesis is still missing for many drugs and chemical
compounds [4,5]. Additionally, there is a lack of large-scale
chemical screens for modifiers of hereditary nephropathies. This is mainly due to the very laborious and time-consuming
testing of substance specific effects on normal and abnormal
organogenesis using standard experimental setups, thus
hampering investigations on a larger scale. Additionally, in vivo
imaging of kidney development in a spatiotemporal context is
not feasible in rodents, necessitating animal models that are
experimentally
more
accessible. Furthermore,
dedicated
imaging techniques are required which enable the in vivo
visualization of developing organs and tissues on a larger
scale. function of the zebrafish pronephros can aid in the
understanding of the role of genes mutated in kidney disease,
or the impact of compounds on renal development and function
in humans [17]. Thus, the combination of this in vivo model
system with automated imaging technologies could serve as a
tool for the large scale analysis of kidney phenotypes. However, to our current knowledge, a screening platform
compatible with in vivo imaging of zebrafish larval kidneys has
not been described yet. Here, we delineate the development of an automated HCS
compatible imaging pipeline designed for live imaging of
zebrafish kidneys in chemical screening scenarios. Using a
custom designed orientation tool, embryos could be accurately
positioned in wells of microtiter plates allowing consistent
imaging of dorsal views of the pronephros. Subsequent
automated imaging was performed on a standard widefield
screening microscope and a data handling and visualization
pipeline was developed. A pilotscreen for morphological kidney
abnormalities was performed using a subset of potentially
nephrotoxic drugs applied to larvae of the Tg(wt1b:EGFP)
transgenic line in which the developing pronephros is
highlighted by GFP expression [19]. The obtained in vivo data
was cross-validated by histological analysis. In addition, we
demonstrate that the established microscopy platform can also
be utilized for genetic disease models. The
development
of
high
content
screening
(HCS)
technologies has had a major impact on biomedical and
pharmaceutical research, as these platforms are suitable for a
wide range of large-scale investigations using in vitro and in
vivo model systems [6]. Due to its small size and various other
experimental advantages, the zebrafish can be readily
employed in large scale in vivo assays. Automated Screening Platform for Zebrafish Kidneys Moreover, the high
degree of anatomical and physiological homology to higher
vertebrates renders it a relevant model for biomedical research
[7]. Thus, the zebrafish has emerged as the main vertebrate
model system for whole organism screening experiments. Consequently, the zebrafish has been successfully used in
chemical, toxicological, behavioral and genetic screening
experiments [4,78910–11]. Moreover, its transparency in
combination with the wealth of mutant and transgenic zebrafish
strains available facilitates the large scale analysis of tissue-
specific phenotypes. This includes, for example, the search for
anti-inflammatory, anti-angiogenic or neuroactive compounds
[1213–14]. Fish keeping and embryo handling Adult zebrafish of the Tg(wt1b:EGFP) transgenic line [19]
were maintained according to reference [20]. Eggs were
collected
from
pairwise
and
batch
crossings. The
developmental stage of embryos was determined as previously
described [21]. Embryos were raised in fish water at 28°C. At
24 hpf embryos were enzymatically dechorionated using 10
mg/ml Pronase. Embryos were transferred to a beaker, washed
twice with 400 ml of fish water and transferred into clean petri
dishes [22]. Prior to transferring into agarose coated microtiter
plates, 48 or 72 hpf old larvae were anesthetized using 0.03%
tricaine. Ethics statement All zebrafish husbandry and experimental procedures were
performed in accordance with the German animal protection
standards and were approved by the Government of Baden-
Württemberg,
Regierungspräsidium
Karlsruhe,
Germany
(Aktenzeichen 35-9185.64). Despite its widespread usage in large scale experiments, it
remains challenging to fully exploit the advantages of this
model system in HCS experiments. Zebrafish embryos are
largely incompatible with standard HCS protocols developed
for other model systems, one of the reasons being that their
relatively large size and complex three-dimensional shape
often lead to random positioning and orientation of embryos
within wells of microtiter plates. This especially hampers
reproducibility and detailed visualization of cell or tissue
morphology, as well as developmental processes. Whilst
several studies and recent technological advances have tried
to address these challenges, they often require sophisticated
technical setups [15] or are incompatible with chemical
screening [16]. Thus, there remains a demand of easy-to-use
screening protocols and dedicated tools which can facilitate
performing complex zebrafish HCS assays. Introduction preterm newborns with ongoing nephrogenesis can interfere
with nephron generation and thus can cause short- and
potentially
long-term
kidney
impairment
[2]. Besides
environmental factors, hereditary nephropathies due to
mutations in genes encoding ciliary proteins are the most
common cause of end-stage renal disease in children due to
polycystic, cystic-dysplastic and/or nephronophthisis like renal
phenotypes [3]. Adverse environmental conditions and genetic influences
can have a major impact on organogenesis. According to
statistical surveys, 22-50% of all pregnant women take drugs
within the first trimenon of gestation, often whilst they are
unaware of their pregnancy [1]. However, nephrotoxic
medication taken by pregnant women or administered to December 2013 | Volume 8 | Issue 12 | e82137 1 PLOS ONE | www.plosone.org Automated Screening Platform for Zebrafish Kidneys Morpholino injections Antisense splice blocking morpholino oligonucleotides (Gene
Tools, LLC, Philomath, USA) were designed against the exon1-
intron1 and exon2 –intron2 boundary of zebrafish ift172 gene,
and ift80 morpholino has been previously published [23]. Morpholinos were injected into 1-cell stage embryos at 500 nM
concentration. Both ift172 morpholinos resulted in similar
phenotypes comparable to the phenotype previously published
[2425–26]. A standard control oligonucleotide was used as
control. Morpholino sequences: ift80 splice blocking MO: 5’-
AGGTGTATGTGGAACCTGTGATAAG-3’, ift172 Exon2 splice
donor
blocking
MO:
5’- Data handling, deconvolution and visualization Data handling, generation of multilayer tiffs and generation of
maximum projections were carried out using custom written
Perl scripts and Fiji [28] macros available on request. Fluorescence z-stacks were deconvolved with Huygens
Professional deconvolution software (SVI, Hilversum, The
Netherlands) using a theoretical point spread function based on
microscope parameters. Batch deconvolution was carried out
on a workstation with 24 CPU cores and 64 Gigabyte of
memory. Cropping of images and generation of overview
images was carried out using a Fiji macro (Macro S1). In brief,
maximum projection images were duplicated, automatically
thresholded and the resulting binary images were eroded. The
position of kidneys was detected by measuring the center of
mass. The corresponding coordinates were restored on original
images, a bounding box was defined and images were cropped
accordingly. Cropped images were loaded into a stack and
overview images were generated using the ‘Make Montage’
function of Fiji. ACGAGATGAGAGCTTACTTTTGGGT-3’, ift172 Exon1 splice
blocking
MO:
5’-ACGCTGTCAATATTTTACCTGAGGC-3’,
Standard
control
(ctr)
oligonucleotide:
5’-
CCTCTTACCTCAGTTACAATTTATA-3’. Drug treatment of embryos A subset of certain drug classes was chosen for which an
adverse effect on the developing kidney had been described in
animal and/or human studies [2]. To evaluate concentration-
dependent toxicity, 5 different concentrations of each drug (2.5
mM, 5 mM, 10 mM, 20 mM, 40 mM) were tested. 24 hpf
dechorionated embryos were transferred to 6-well-plates and
treated with the following drugs dissolved in E3 solution with
0.003% 1-pheny-2-thiourea (PTU, Alfa Aesar, Karlsruhe, In the zebrafish larva, the functional pronephros comprises
only 2 nephrons with fused glomeruli located ventrally to the
dorsal aorta. Interestingly, despite tremendous differences in
nephron number, the composition of a single nephron shows
great homology at the cellular and molecular level between
human and zebrafish [17,18]. Additionally, kidney development
and function largely depend on the same orthologous genes for
all vertebrate kidneys. Therefore, studying formation and PLOS ONE | www.plosone.org December 2013 | Volume 8 | Issue 12 | e82137 2 Automated Screening Platform for Zebrafish Kidneys Germany): penicillin G potassium salt (AppliChem, Darmstadt,
Germany), ampicillin sodium salt (AppliChem, Darmstadt,
Germany), gentamicin sulfate (Sigma-Aldrich, St. Louis, USA),
kanamycin sulfate (AppliChem, Darmstadt, Germany), captopril
(CalBiochem, Darmstadt, Germany), losartan potassium salt
(Molekula,
Gillingham,
Dorset,
United
Kingdom),
acetaminophen (Caesar und Loretz, Hilden, Germany),
indomethacin sodium salt (AppliChem, Darmstadt, Germany). Treatment period was 24 hours. For indomethacin, lower
concentrations (0.01 mM, 0.025 mM, 0.05 mM, 0.075 mM and
0.1 mM) had to be applied due to 100% lethality rates at higher
concentrations. 2-4 repeats of each experiment were
performed. Total number of embryos that underwent drug
treatment is shown in Table S1. The pH was adjusted for each
experiment. Following drug treatment, the number of dead
larvae was assessed and the living larvae were transferred to
0.003% PTU containing E3 solution. For imaging studies,
0.03% tricaine was added to the medium. 4°C. Embryos were transferred in 100 µl fish water containing
PTU and tricaine (see above) and manually arrayed and
oriented under a stereomicroscope. To aid in positioning of
regions of interest within the grid-based and fixed field views of
the Scan^R system, embryos were positioned in such a way
that all yolk sacs were approximately at the same position
within cavities using features of the well plate as guidelines. Image acquisition Embryos were imaged on a standard Scan^R high-content
screening microscope (Olympus, Hamburg, Germany) [27] as
previously described [16,22]. Data was acquired using 33 z-
slices (Δz = 15μm) per embryo and channel using a 4x (N.A. =
0.13) objective. Integration times were fixed (80 ms for GFP). Imaging times were approximately 1 hour for a full 96 well
plate. For each experimental plate the A1 positions of the
imaging grid was re-centered to compensate for minor
differences in positioning of embryos. Generation of a 96 well template tool The tool is made from brass and consists of a base plate
with 96 perpendicular pins arranged in a certain way to match
the positions of the wells of a microtiter plate. The tool was
produced by CNC milling (Datron, M7). In a first step, the base
plate with the dimensions of 140 mm x 100 mm x 21 mm was
milled by a four flute end mill (20 mm). Secondly, the array of
96 pins with 11 mm height and rectangular footprint (5.9 mm x
1 mm) was created by partly thinning the base plate down to
9 mm between the pins (Four Flute End Mill, 6 mm and 3 mm,
respectively). Finally, the ends of the 96 pins were tapered to a
conical tip with 60° by utilizing a standard engraving tool. Analysis of morphological and pronephric phenotypes Analysis of morphological and pronephric phenotypes
Overall morphology was scored on a Leica MZ10 F
stereomicroscope (Leica Microsystems, Wetzlar, Germany). Lethality and pericardial/yolk sac edema were rated for each
experiment. Blinded analysis of tubular and glomerular
phenotypes was performed by SS and JHW on maximum
projections of deconvolved z-stacks. Using ImageJ, 2
parameters were manually measured for tubular structures: i)
maximum distance between the tubules and ii) angle between
neck segment and proximal convoluted tubule [18] being visible
in Tg(wt1b:EGFP) zebrafish. For description of glomerular
changes, the distance between the glomeruli was determined
and glomeruli were classified as either normal or showing
glomerular malformation judged by the glomerular area and
structure. Heatmaps were generated using matrix2png [29]. Standard positioning of embryos for chemical
screening The visualization of bilateral symmetric organs of zebrafish
embryos usually demands dorsal or ventral views. However,
consistent large scale imaging of zebrafish embryos remains
challenging in automated screening experiments, as stable and
reproducible positioning is complicated by the size and
complex three-dimensional shape of embryos. The screening system used in this study stores data as
single tiff files in one folder per experimental plate. To reduce
file number and thus facilitate subsequent data handling and
analysis tasks, multilayer tiff files were generated for each
imaging position and channel. Spatial widefield data usually
suffers from out-of-focus blur thus reducing overall image
quality [30]. To restore images, fluorescent datasets were
batch deconvolved with Huygens Professional software using a
theoretical point spread function [16]. Subsequently, maximum
projection images were generated from deconvolved z-stacks
(Figure 2E). The positions of larval kidneys were automatically
detected within maximum projections of deconvolved data
using the center of mass of the corresponding binary image
after automatic thresholding. To restrict image data to the
pronephric region and remove unnecessary areas, a bounding
box was defined around the center of mass and images were
cropped accordingly (Figure 2F). Subsequently, overview
images
were
generated
from
cropped
kidney
images
representing all kidneys in one 96-well plate, allowing for rapid
manual assessment of morphological phenotypes (Figure 2G). In summary, this pipeline allows consistent imaging and rapid
evaluation of gross morphological abnormalities of the
developing zebrafish kidney after compound treatment or in
genetic screens. It can also be easily adapted for the analysis
of other tissues and organs that require consistent imaging. To enable a simplified handling and precise positioning of
zebrafish larvae and to achieve consistent visualization of
tissues in high content screening scenarios, we have
developed a tool to create agarose molds in a standard
microtiter plate. The tool allows the preparation of agarose
coated 96 well plates in a single replication step. The tool
consists of a base plate with 96 rectangular pins, whose
positions exactly match the centers of wells of standard and
commercially available 96 well microtiter plates (Figure 1A, B). The pins end with a keel shaped geometry, which was
previously shown to be suitable for accurate ventral positioning
of zebrafish larvae [16]. Statistical analysis Data were evaluated using IBM® SPSS® Statistics Version
21. For lethality and edema rates and glomerular fusion and
malformation, statistical analysis was performed by Chi-square
test. Datasets of low sample sizes were additionally tested
using Fisher’s exact test. For tubular angle and distance,
means among treatment groups were compared using ANOVA
with Bonferroni correction for multiple comparisons as a post-
hoc test. Significance was defined as p<0.05. Pipeline for automated pronephros imaging To visualize and score renal phenotypes, we developed a
protocol for automated imaging of dorsal views of zebrafish
larval kidneys (Figure 2A). Prior to imaging, compound treated
or microinjected embryos of the Tg(wt1b:EGFP) stable
transgenic line were raised to the desired developmental stage
(48 or 72 hpf) and then transferred into microtiter plates
containing agarose cavities as described above (Figure 2B, C). To automatically image zebrafish kidneys, larvae were
manually positioned and oriented in the agarose cavities and
subsequently imaged on a standard widefield HCS microscope. To ensure capture of entire organs and compensate for minor
variations in z-positioning, each larva was acquired using z-
stacks with 33 slices in the bright field and GFP channel
(Figure 2D). The cavities in the plate allow larvae to be
positioned with high enough accuracy, so that regions of
interest (e.g. pronephros) are located within the limited field of
view for all plated embryos. Thus, the tool permits the organ
and
tissue
specific
screening
on
standard
screening
microscopes using a fixed field of view for all wells, without the
necessity of additional software modules for automatic
detection and centering the region of interests [16]. Histological analysis Histological analysis device allows generation of molds in each well independently,
thus avoiding cross-contamination, which was a major
limitation of the previous design. Furthermore, the restriction to
single wells and the depths of the cavities drastically minimizes
movement of surrounding medium leading to a massively
improved stability of orientation of embryos in comparison to
the silicone template. Thus, plates with oriented embryos and
larvae could be used in combination with automated plate
handling and stacking systems. Larvae were fixed in 4% paraformaldehyde overnight at 4°C. Samples were dehydrated through an ethanol series and
processed for embedding in Paraffin (Surgipath® Paraplast®,
Leica Biosystems, Wetzlar, Germany). 3 μm sections were cut
using a Leica RM 2165 microtome (Leica Microsystems,
Nussloch, Germany). Sections were deparaffinized in xylene
and rehydrated through graded washes of ethanol in water
before staining with hematoxylin and eosin. The stained
sections were imaged with a Leica DMI4000 B microscope
equipped with a Leica DFC320 digital camera. Automated Screening Platform for Zebrafish Kidneys Automated Screening Platform for Zebrafish Kidneys Preparation of agarose molds in microtiter plates Preparation of agarose molds in microtiter plates
70 µl of 1% agarose in fish water was added to each well of
a 96 well microtiter plate (Cat.-No. 655180, Greiner,
Frickenhausen, Germany) using a multi-channel pipette. The
agarose coated well plates were pre-cooled at room
temperature for one minute. To generate grooves the brass
tool was inserted, while adjusting the penetration depth of pins
using spacers. After solidification the tool was carefully
removed and plates were optionally stored in plastic bags at December 2013 | Volume 8 | Issue 12 | e82137 PLOS ONE | www.plosone.org 3 Standard positioning of embryos for chemical
screening The tool was fabricated out of a solid
block of brass using CNC milling (Figure 1A, B), giving rise to a
precise work piece with identical xyz-dimensions of each pin
allowing the generation of deep keel-shaped cavities in wells of
a 96-well plate filled with agarose (Figure 1C). Such prepared
plates can be readily used to manually position specimen
enabling the subsequent automated acquisition of dorsal views
using inverted screening microscope systems (Figure 1D). using inverted screening microscope systems (Figure 1D). We recently demonstrated an alternative protocol for
automated dorsal imaging of oriented larvae using a silicone
tool to generate an array of 96 agarose molds [16]. However,
while plates generated with this tool can be employed for the
consistent automated imaging of tissues, the design prevents
utilization in chemical or drug screening applications. The novel December 2013 | Volume 8 | Issue 12 | e82137 PLOS ONE | www.plosone.org 4 4 Automated Screening Platform for Zebrafish Kidneys Figure 1. Standardized orientation of zebrafish embryos. (A,B) Photographs of the brass tool for the simultaneous generation
of agarose grooves within 96 well microtiter plates: (A) top view and (B) tilted view. For dimensions of the plate see Materials and
Methods section. (C) Schematic depiction of a single well with a ventrally oriented embryo within an agarose cavity. Drawing is not
to scale. (D) Illustrative example of aligned and oriented embryos. Shown are dorsal views of 48 hpf embryos acquired using a 2.5x
objective on an inverted wide field screening microscope. doi: 10.1371/journal.pone.0082137.g001 Figure 1. Standardized orientation of zebrafish embryos. (A,B) Photographs of the brass tool for the simultaneous generation
of agarose grooves within 96 well microtiter plates: (A) top view and (B) tilted view. For dimensions of the plate see Materials and
Methods section. (C) Schematic depiction of a single well with a ventrally oriented embryo within an agarose cavity. Drawing is not
to scale. (D) Illustrative example of aligned and oriented embryos. Shown are dorsal views of 48 hpf embryos acquired using a 2.5x
objective on an inverted wide field screening microscope. doi: 10.1371/journal.pone.0082137.g001 A pilot screen for drug-related effects on kidney
development dependent effects on overall survival rates, edema formation
and pronephric phenotypes. Detailed results are listed in Table
S1-S3. Color coded overview maps were also generated
(Figure 3I-M). To validate phenotypic alterations observed in
the transgenic model and thus confirm the utility of the
proposed screening pipeline, histological analysis of glomerular
and tubular cross-sections of 48 hpf larvae treated at the
highest non-lethal concentration was carried out (Figure S1). The pipeline was subsequently evaluated in a pilot screen to
investigate the impact of potentially nephrotoxic drugs on the
development of the zebrafish pronephros. To this end, a subset
of drugs from different classes was chosen for which an
adverse effect on the developing kidney was previously
identified in animal studies and/or human observations [2]. Dechorionated 24 hpf old embryos were treated with 8 different
drugs in increasing concentrations for 24 hours. Following drug
treatment, live larvae were imaged and data was visualized as
described above. Impact of tested compounds on kidney development Impact of tested compounds on kidney development
In humans, due to the putative absence of fetal toxicity at
therapeutic doses, penicillin antibiotics are widely prescribed to
pregnant women and frequently administered to preterm
newborns [31]. In our study, penicillin G potassium salt
administration increased lethality rates dose-dependently
(Figure 3I). Concomitantly, minor pronephric alterations were
observed including incompletely fused glomeruli (Figure 3B). On the other hand, ampicillin sodium salt did not cause higher
lethality, increased edema rates or major phenotypic renal
alterations (Figure 3C, I-M). Cross-sections of larvae treated
with penicillin G or ampicillin did not show major glomerular or
tubular alterations when compared to untreated control larvae
(Figure
S1A-C),
thus
confirming
results
obtained
by
fluorescence microscopy. Taken together, this indicates only To score lethality rates and development of pericardial and
yolk sac edema, treated larvae were examined on a stereo
microscope. The detailed results are listed in Table S1. To objectively quantify morphological abnormalities of the
pronephros following drug treatment, glomerular and tubular
alterations were discriminated in the Tg(wt1b:EGFP) transgenic
line. Glomerular alterations were subdivided into (i.) glomerular
malformation indicated by abnormal or reduced glomerular
shape and area, and (ii.) incomplete glomerular fusion
representing
aberrant
pronephros
development. Tubular
parameters were classified into (i.) the angle between the
tubular neck segment and the proximal convoluted tubule and
(ii.) variations in the maximum distance between the 2 tubular
systems (Figure 3A). Several drugs showed concentration PLOS ONE | www.plosone.org December 2013 | Volume 8 | Issue 12 | e82137 5 Automated Screening Platform for Zebrafish Kidneys Figure 2. Overview of workflow for the automated imaging of the developing zebrafish pronephros. (A) Overview of the
workflow for screening larval kidneys. The flowchart illustrates the different steps carried out to obtain overview images of kidneys. (B) Initial compound treatment or microinjection of embryos prior to sample preparation and imaging. (C) Schematic illustrating the
transfer of embryos into agarose coated microtiter plates, and alignment and orientation of embryos. (D-G) Acquisition and
processing of image Data. D to F show data of the same embryo. (D) Automated acquisition of z-stacks (33 z-slices, dZ=15µm) on
an inverted widefield screening microscope. (E) Deblurring of images using deconvolution. Shown are maximum projections of z-
stacks of raw data (left panel) and deconvolved data (right panel). (F) Automated detection and cropping of the kidney region. The
red square indicates the position and dimensions of the cropped region. Impact of tested compounds on kidney development (G) Automated generation of overview images for quick
assessment of overall morphological changes. Indomethacin skeletal formula in (A) taken from (http://en.wikipedia.org/wiki/
File:Indometacin_skeletal.svg). d i 10 1371/j
l
0082137
002 Figure 2. Overview of workflow for the automated imaging of the developing zebrafish pronephros. (A) Overview of the
workflow for screening larval kidneys. The flowchart illustrates the different steps carried out to obtain overview images of kidneys. (B) Initial compound treatment or microinjection of embryos prior to sample preparation and imaging. (C) Schematic illustrating the
transfer of embryos into agarose coated microtiter plates, and alignment and orientation of embryos. (D-G) Acquisition and
processing of image Data. D to F show data of the same embryo. (D) Automated acquisition of z-stacks (33 z-slices, dZ=15µm) on
an inverted widefield screening microscope. (E) Deblurring of images using deconvolution. Shown are maximum projections of z-
stacks of raw data (left panel) and deconvolved data (right panel). (F) Automated detection and cropping of the kidney region. The
red square indicates the position and dimensions of the cropped region. (G) Automated generation of overview images for quick
assessment of overall morphological changes. Indomethacin skeletal formula in (A) taken from (http://en.wikipedia.org/wiki/
File:Indometacin_skeletal.svg). doi: 10 1371/journal pone 0082137 g002 minor effects of the 2 β-lactam antibiotics on pronephros
development. only a minor increase in lethality rates without significant
effects on edema formation (Figure 3I, J). Nevertheless,
glomerular malformation and incomplete glomerular fusion
were found for higher drug concentrations. Tubular angle was
slightly widened at higher doses (Figure 3D, M). Histological
analysis of gentamicin-treated larvae revealed no gross
morphological abnormalities, although a mild rarefication of Aminoglycosides,
although
not
recommended
during
pregnancy [31], are often used for treatment of neonatal
sepsis,
even
in
premature
newborns
with
on-going
nephrogenesis. However, serum drug concentrations can be
monitored to minimize the risk of renal and auditory toxicity
[32]. In our study, gentamicin sulfate administration caused PLOS ONE | www.plosone.org PLOS ONE | www.plosone.org December 2013 | Volume 8 | Issue 12 | e82137 6 Automated Screening Platform for Zebrafish Kidneys Figure 3. Overview of compound concentration-dependent pronephric phenotypes. Illustrative examples of pronephroi of a
(A) non-treated embryo, and after treatment with (B) 20 mM penicillin, (C) 40 mM ampicillin, (D) 40 mM gentamicin, (E) 40 mM
kanamycin, (F) 40 mM acetaminophen, (G) 40 mM captopril, (H) 10 mM losartan. For examples of phenotypes after Indomethacin
treatment see Figure 4A-E. Impact of tested compounds on kidney development Arrow and arrowheads in (A) indicate the different morphological parameters of the pronephros scored
to evaluate compound effect on the developing kidney. Arrow: fused glomeruli; Arrowhead: angle between the neck segment and
the proximal convoluted tubule. (I-M) Heatmaps showing (I) lethality rates, (J) edema rates and (K-M) changes in morphological
parameters of the pronephros. In detail, (K) incomplete glomerular fusion, (L) glomerular malformation and (M) tubular angle. For
further details see Materials and Methods and Tables S1-S3. Colour codes indicate the percentage of embryos (I-L) with particular
phenotype, or the angle between neck segment and proximal convoluted tubule (M) as indicated by the colour coded legend. Grey
squares indicate missing data points. Concentration ranges used are indicated above the heatmaps, or below for Indomethacin,
respectively. Abbreviations: penicillin (Pen), ampicillin (Amp), gentamicin (Gen), kanamycin (Kan), acetaminophen (Ace), captopril
(Cap), losartan (Los) and indomethacin (Ind). *p<0.05, **p<0.001. doi: 10.1371/journal.pone.0082137.g003 Figure 3. Overview of compound concentration-dependent pronephric phenotypes. Illustrative examples of pronephroi of a
(A) non-treated embryo, and after treatment with (B) 20 mM penicillin, (C) 40 mM ampicillin, (D) 40 mM gentamicin, (E) 40 mM
kanamycin, (F) 40 mM acetaminophen, (G) 40 mM captopril, (H) 10 mM losartan. For examples of phenotypes after Indomethacin
treatment see Figure 4A-E. Arrow and arrowheads in (A) indicate the different morphological parameters of the pronephros scored
to evaluate compound effect on the developing kidney. Arrow: fused glomeruli; Arrowhead: angle between the neck segment and
the proximal convoluted tubule. (I-M) Heatmaps showing (I) lethality rates, (J) edema rates and (K-M) changes in morphological
parameters of the pronephros. In detail, (K) incomplete glomerular fusion, (L) glomerular malformation and (M) tubular angle. For
further details see Materials and Methods and Tables S1-S3. Colour codes indicate the percentage of embryos (I-L) with particular
phenotype, or the angle between neck segment and proximal convoluted tubule (M) as indicated by the colour coded legend. Grey
squares indicate missing data points. Concentration ranges used are indicated above the heatmaps, or below for Indomethacin,
respectively. Abbreviations: penicillin (Pen), ampicillin (Amp), gentamicin (Gen), kanamycin (Kan), acetaminophen (Ace), captopril
(Cap), losartan (Los) and indomethacin (Ind). *p<0.05, **p<0.001. doi: 10.1371/journal.pone.0082137.g003 Figure 3. Overview of compound concentration-dependent pronephric phenotypes. Automated microscopy screening of genetic kidney
disease models Additionally, captopril and losartan treated larvae displayed
slightly altered glomerular structure in histological sections that
appeared less dense compared to controls (Figure S1G,
H).This data is partially consistent with animal studies showing
renal abnormalities after treatment with ACE inhibitors or
angiotensin receptors [4748–49]. g
y
y
[
]
Splice blocking morpholinos for ift80 and ift172 were
designed as described in the Methods section. By using the
standard positioning tool as described above, automated
imaging
of
dorsally
positioned
morpholino-injected
Tg(wt1b:EGFP) zebrafish (3 dpf) was performed in 96 well
plates (Figure 5A). Microscopy revealed a ventral curvature of
the tail (Figure 5B-D) affecting approximately 90% of all
morpholino
injected
embryos
and
consistent
with
the
phenotype previously described for ift80 morphants [23]. Morphological
alterations
in
fluorescence
microscopy
predominantly consisted of large cystic glomeruli (Figure 5E-G)
that were consistently reproducible [23], while embryos treated
with
standard
morpholino
showed
normal
glomerular
morphology. Cross-sections
of
both
ift80-
and
ift172-
morpholino injected Tg(wt1b:EGFP) zebrafish confirmed the
formation of large pronephric cysts (Figure S2A-C). Tubular
dilatation and epithelial flattening was observed both in
fluorescence images (Figure 5F, G) and histological sections
(Figure S2B, C). This further demonstrates that our pipeline is
suitable for large scale therapeutic screening investigations in
genetic models of renal disease such as ciliopathies. However,
the applicability largely depends on the morphological
phenotypes as severe malformations impair position accuracy
within cavities. NSAIDs are widely used for closure of patent ductus
arteriosus in preterms and are administered during pregnancy
for prevention and treatment of toxemia, polyhydramnions and
premature birth. However, exposure to NSAIDs during
pregnancy can cause hypoperfusion of the fetal kidneys and
acute renal failure in newborns with cystic changes of
developing nephrons and long-term renal dysfunction [2,50]. In
our study, concentrations of the non-selective COX1/COX2
inhibitor indomethacin had to be lowered due to 100% lethality
at higher doses. Strikingly, even at drastically lower
concentrations there was a severe phenotype (Figure 4A-E). This included concentration dependent increases in edema
formation
and
lethality
(Figure
4D,
F). Fluorescence
microscopy (Figure 4A, E) revealed significant concentration
dependent
increases
in
glomerular
malformation
and
incomplete glomerular fusion (Figure 4G). Furthermore, tubular
angles widened and tubular distances slightly shortened
(Figure 4H). Severe renal phenotypes were also confirmed in
histological sections of indomethacin treated larvae. Here, no
regular glomerulus was detectable ventrally to the dorsal aorta
and laterally seen glomerular structures appeared strongly
malformed or were not identifiable. Automated microscopy screening of genetic kidney
disease models been considered safe [38]. Its hepatotoxicity at high doses is
well described [39] and has recently been investigated in
zebrafish [40]. In addition, animal data further revealed fetal
kidney damage following acetaminophen administration to
pregnant rats [41]. In our study, acetaminophen caused
concentration dependent significant alterations of pronephros
morphology and an increase in edema formation, whereas
lethality
rates
remained
unchanged
(Figure
3F,
I-M). Histological sections confirmed severe renal phenotypes
following acetaminophen administration. No fused glomerulus
was detectable ventrally to the dorsal aorta and glomerular
structures appeared strongly malformed. In addition, tubular
epithelium was flattened (Figure S1F). These results match
previously published data showing dose-, duration- and onset-
dependent changes in pronephros morphology following
acetaminophen administration in zebrafish larvae [42]. Beyond performing toxicological screens for kidney damage,
the presented automated microscopy pipeline can be utilized in
the analysis of genetic disease models. Gene-knockdown or
knock-out
models
can
potentially
be
used
for
HCS
investigations for the search of therapeutic strategies for
hereditary kidney diseases. To test the utility of the developed
pipeline, we focused on cilia-associated human genetic
disorders. Intraflagellar transport (IFT) constitutes the bidirectional
transport of protein complexes along axonemal microtubules. IFT plays an essential role in the assembly and function of cilia
and flagella by contributing to cell motility, sensory perception
and cilium-based signaling [51,52]. IFT80 and IFT172 both are
members of the IFT-B subcomplex [53] and while IFT80
mutations in humans have been identified to cause Jeune
asphyxiating thoracic dystrophy [23], a congenital ciliary
chondrodysplasia condition associated with renal disease in
approximately 20% of cases [54], no human mutations in
IFT172 have been identified to date. However, abrogation of
Ift172 function in mice leads to a VACTERL-like phenotype
including renal malformations [55], indicating that IFT172 plays
an important role for kidney development in mammals. In
zebrafish, the insertional mutant line ift172hi2211Tg exhibits
glomerular cysts and a ventral body curvature [56]. Intake of ACE inhibitors and angiotensin receptor blockers
during pregnancy has been associated with fetopathies
including renal pathologies in humans [4344–45] and other
mammals [46]. In our study, captopril at 40 mM significantly
increased edema formation and induced concentration
dependent alterations in glomerular and tubular parameters
(Figure 3G, I-M). Losartan increased lethality rates and
glomerular and tubular parameters at higher concentrations,
while edema rates were unchanged (Figure 3H, I-M). Impact of tested compounds on kidney development Illustrative examples of pronephroi of a
(A) non-treated embryo, and after treatment with (B) 20 mM penicillin, (C) 40 mM ampicillin, (D) 40 mM gentamicin, (E) 40 mM
kanamycin, (F) 40 mM acetaminophen, (G) 40 mM captopril, (H) 10 mM losartan. For examples of phenotypes after Indomethacin
treatment see Figure 4A-E. Arrow and arrowheads in (A) indicate the different morphological parameters of the pronephros scored
to evaluate compound effect on the developing kidney. Arrow: fused glomeruli; Arrowhead: angle between the neck segment and
the proximal convoluted tubule. (I-M) Heatmaps showing (I) lethality rates, (J) edema rates and (K-M) changes in morphological
parameters of the pronephros. In detail, (K) incomplete glomerular fusion, (L) glomerular malformation and (M) tubular angle. For
further details see Materials and Methods and Tables S1-S3. Colour codes indicate the percentage of embryos (I-L) with particular
phenotype, or the angle between neck segment and proximal convoluted tubule (M) as indicated by the colour coded legend. Grey
squares indicate missing data points. Concentration ranges used are indicated above the heatmaps, or below for Indomethacin,
respectively. Abbreviations: penicillin (Pen), ampicillin (Amp), gentamicin (Gen), kanamycin (Kan), acetaminophen (Ace), captopril
(Cap), losartan (Los) and indomethacin (Ind). *p<0.05, **p<0.001. doi: 10.1371/journal.pone.0082137.g003 capillary loops could be seen (Figure S1D). Kanamycin caused
a concentration-dependent increase in lethality and edema
formation (Figure 3I, J). However, glomerular and tubular
parameters remained unaltered (Figure 3E). Concordantly, no
major glomerular or tubular alterations were observed in
histological
sections
of
larvae
following
kanamycin
administration (Figure S1E). In other studies, microinjection of
gentamicin into the cardiac venous sinus led to acute renal
failure [33]. As only minor effects of gentamicin were observed in our study, it suggests that this may have been due to poorer
penetration into inner organs. Several human and animal
studies report on aminoglycoside-induced glomerular and
tubular damage in pre- and at-term newborns [3435–36]. Substance-specific differences in the degree of ototoxic and
nephrotoxic side effects among various aminoglycosides are
well known [37]. In humans, the intake of acetaminophen at therapeutic doses
during gestation and administration to preterm newborns has PLOS ONE | www.plosone.org PLOS ONE | www.plosone.org 7 December 2013 | Volume 8 | Issue 12 | e82137 December 2013 | Volume 8 | Issue 12 | e82137 Automated Screening Platform for Zebrafish Kidneys Automated microscopy screening of genetic kidney
disease models Additionally, pronephric
tubular epithelium appeared flattened (Figure S1I). Thus, our
data in larval zebrafish matches previously published data from
other animal models confirming severe renal side effects of
indomethacin on kidney development during nephrogenesis
[50]. Conclusions Here, we demonstrate the development of an automated
screening pipeline for imaging developing kidneys in the
zebrafish larvae. This novel methodology allows for the
consistent acquisition of dorsal views of pronephric kidneys on
a standard inverted screening microscope. The platform can PLOS ONE | www.plosone.org December 2013 | Volume 8 | Issue 12 | e82137 8 Automated Screening Platform for Zebrafish Kidneys Figure 4. Impairment of pronephros development upon indomethacin treatment. (A) Overview of pronephric alterations in
zebrafish larvae (50 hpf) following indomethacin administration for 24 hours. Row 1 shows control embryos, rows 2-6 zebrafish
embryos following indomethacin administration in increasing concentrations (row 2, 0.01 mM; row 3, 0.025 mM; row 4, 0.05 mM;
row 5, 0.075 mM and row 6, 0.1 mM). (B-E) Comparison of (B-C) 50 hpf control larva and (D-E) indomethacin (0.1 mM) treated
larva. (D) Brightfield image shows edema formation following indomethacin administration. (E) Fluorescence image showing
nephron (glomerular and tubular) malformation. (F) Quantification of lethality rates and edema formation following indomethacin
administration. (G) Concentration-dependent increases in glomerular malformation and decreases in glomerular fusion rates
following indomethacin administration. (H) Widened tubular angles between neck segment and proximal convoluted tubule following
indomethacin administration. Data are shown as mean ± SD. *p<0.05, **p<0.001. doi: 10.1371/journal.pone.0082137.g004 Figure 4. Impairment of pronephros development upon indomethacin treatment. (A) Overview of pronephric alterations in
zebrafish larvae (50 hpf) following indomethacin administration for 24 hours. Row 1 shows control embryos, rows 2-6 zebrafish
embryos following indomethacin administration in increasing concentrations (row 2, 0.01 mM; row 3, 0.025 mM; row 4, 0.05 mM;
row 5, 0.075 mM and row 6, 0.1 mM). (B-E) Comparison of (B-C) 50 hpf control larva and (D-E) indomethacin (0.1 mM) treated
larva. (D) Brightfield image shows edema formation following indomethacin administration. (E) Fluorescence image showing
nephron (glomerular and tubular) malformation. (F) Quantification of lethality rates and edema formation following indomethacin
administration. (G) Concentration-dependent increases in glomerular malformation and decreases in glomerular fusion rates
following indomethacin administration. (H) Widened tubular angles between neck segment and proximal convoluted tubule following
indomethacin administration. Data are shown as mean ± SD. *p<0.05, **p<0.001. doi: 10.1371/journal.pone.0082137.g004 the imaging protocol is easy-to-use and can be readily modified
for studying other organ systems or tissues such as the brain
region. Furthermore, in combination with HCS software tools
which enable automated feature of interest detection, it can be serve as a convenient tool in kidney research, e.g. Supporting Information Figure S1. Cross-sections of pronephric regions after
compound exposure. Shown are glomerular (upper panels)
and tubular (lower panels) sections at 48 hours post
fertilization. (A) control, (B) penicillin (20 mM), (C) ampicillin (40
mM), (D) gentamicin (40 mM), (E) kanamycin (40 mM), (F)
acetaminophen (40 mM), (G) captopril (40 mM), (H) losartan
(10 mM) and (I) indomethacin (0.75 mM) treatment. (TIF) Figure S1. Cross-sections of pronephric regions after
compound exposure. Shown are glomerular (upper panels)
and tubular (lower panels) sections at 48 hours post
fertilization. (A) control, (B) penicillin (20 mM), (C) ampicillin (40
mM), (D) gentamicin (40 mM), (E) kanamycin (40 mM), (F)
acetaminophen (40 mM), (G) captopril (40 mM), (H) losartan
(10 mM) and (I) indomethacin (0.75 mM) treatment. (TIF) The current pipeline only allows for scoring of gross
morphological abnormalities of the pronephros. Therefore, we
validated the impact of treatments on pronephros formation by
histological analysis, which largely confirmed the phenotypes
observed in the transgenic model. This further demonstrates
the utility of the established screening pipeline to score
pronephric phenotypes. Moreover, the histological analysis
revealed additional alterations, such as epithelial flattening,
which are more difficult to score in the fluorescence data, thus
complementing the in vivo approach. Although protocols for
large scale histology experiments exist [57], the fast filtering of
compound libraries by in vivo screening is considerably more
time- and cost effective. Therefore, we propose that in a
genuine large scale experiment, histological analysis is only
carried out as a follow-up experiment in combination with
molecular methods to characterize hits identified in a screen. Figure S2. Cross-sections of pronephric regions after
morpholino injections. Shown are glomerular (upper panels)
and tubular (lower panels) sections at 72 hours post
fertilization. (A) control-MO, (B) ift80-MO and (C) ift172-MO
injection. (TIF) Figure S2. Cross-sections of pronephric regions after
morpholino injections. Shown are glomerular (upper panels)
and tubular (lower panels) sections at 72 hours post
fertilization. (A) control-MO, (B) ift80-MO and (C) ift172-MO
injection. (TIF) Table S1. Lethality and edema formation in zebrafish
larvae following drug treatment. (DOCX) The mode of drug administration imposes another limitation
of the established pipeline, as penetration to inner organs can
be hampered by the chemical and physical properties of the
noxa and the biological barrier of the larval zebrafish skin. This
can be overcome by microinjection of drugs into the blood
stream as described by Hentschel et al. [33]. Conclusions in chemical
studies as a primary screening tool to identify organotoxic
substances or to search for potential therapeutic compounds
that attenuate renal pathology in disease models. Importantly, PLOS ONE | www.plosone.org PLOS ONE | www.plosone.org December 2013 | Volume 8 | Issue 12 | e82137 9 Automated Screening Platform for Zebrafish Kidneys 5. Detection of cystic kidneys after Ift80 and Ift172 knockdown. (A) Automatically generated overview thumbnail i
kidneys of morpholino injected larvae: standard ctr-MO (row 1-2), ift80-MO (row 3-5) and ift172-MO (row 6-8). ypic alterations of zebrafish larvae at 4 dpf after morpholino microinjection; (C) ift80-MO and (D) ift172-MO injected em
glomerular cyst (arrow) and a ventrally curved tail compared to (B) standard ctr-MO injected embryos. (E-G) In-
ation of glomerular cysts at 72 hpf in (F) ift80-MO and (G) ift172-MO injected embryos compared to standard ctr-MO inj
s (E) using the Tg(wt1b:EGFP) transgenic line. Figure 5. Detection of cystic kidneys after Ift80 and Ift172 knockdown. (A) Automatically generated overview thumbnail image
of 96 kidneys of morpholino injected larvae: standard ctr-MO (row 1-2), ift80-MO (row 3-5) and ift172-MO (row 6-8). (B-D)
Phenotypic alterations of zebrafish larvae at 4 dpf after morpholino microinjection; (C) ift80-MO and (D) ift172-MO injected embryos
exhibit glomerular cyst (arrow) and a ventrally curved tail compared to (B) standard ctr-MO injected embryos. (E-G) In-detail
visualisation of glomerular cysts at 72 hpf in (F) ift80-MO and (G) ift172-MO injected embryos compared to standard ctr-MO injected
embryos (E) using the Tg(wt1b:EGFP) transgenic line. doi: 10.1371/journal.pone.0082137.g005 Detection of cystic kidneys after Ift80 and Ift172 knockdown. (A) Autom
dneys of morpholino injected larvae: standard ctr-MO (row 1-2)
ift80-MO Figure 5. Detection of cystic kidneys after Ift80 and Ift172 knockdown. (A) Automatically generated overview thumbnail image
of 96 kidneys of morpholino injected larvae: standard ctr-MO (row 1-2), ift80-MO (row 3-5) and ift172-MO (row 6-8). (B-D)
Phenotypic alterations of zebrafish larvae at 4 dpf after morpholino microinjection; (C) ift80-MO and (D) ift172-MO injected embryos
exhibit glomerular cyst (arrow) and a ventrally curved tail compared to (B) standard ctr-MO injected embryos. (E-G) In-detail
visualisation of glomerular cysts at 72 hpf in (F) ift80-MO and (G) ift172-MO injected embryos compared to standard ctr-MO injected
embryos (E) using the Tg(wt1b:EGFP) transgenic line. Macro S1. Fiji macro to crop images and generate
overview images.
(ZIP) Macro S1. Fiji macro to crop images and generate
overview images. (ZIP) Conclusions doi: 10.1371/journal.pone.0082137.g005 December 2013 | Volume 8 | Issue 12 | e82137 10 PLOS ONE | www.plosone.org Automated Screening Platform for Zebrafish Kidneys be developed or modified to be compatible with automated
imaging assays, respectively [61]. used
for
the
automated
acquisition
of
standardized
multidimensional high resolution datasets. The pilot screen for nephrotoxic drugs during nephrogenesis
revealed
a
concentration
dependent
effect
of
several
compounds on nephrogenesis. Thus, considering the lingering
lack of data for many substances, subsequent large-scale
investigations performed with this screening pipeline might
contribute
to
our
understanding
of
substance-specific
nephrotoxic side effects. Acknowledgements Genetic research over the last years has demonstrated that
the zebrafish pronephros is a valuable model system for the
study of hereditary human nephropathies as abnormalities in
podocyte gene function, renal epithelial primary cilia genes and
renal ion channels and transporters lead to defective
pronephric kidney function in the zebrafish mimicking human
disease [59]. However, screening for disease modulating
compounds in a zebrafish model requires convenient and
accessible protocols. Here, we demonstrate that the developed
imaging pipeline can also be utilized to detect abnormal
phenotypes in genetic disease models. Thus, it could serve as
a platform for prospective high-content drug screening
experiments. We thank Sebastian Hötzel for fish care, J. Bohn for CNC
milling, B. Rodenbeck for technical assistance and Dr
Christoph Englert (Leibniz Institute for Age Research-Fritz
Lipmann Institute, Jena, Germany) for kindly providing the
Tg(wt1b:EGFP) transgenic line. We are grateful to Dr Jens
Fahrenberg (Innovation Department, Karlsruhe Institute of
Technology, Germany) for general support. We thank Sundeep
Dhillon for critically reading the manuscript. Supporting Information Duration of drug
treatment and its transferability to the human situation is
another limitation of the pilot screen in zebrafish. In zebrafish,
glomerular filtration starts around 48 hpf and the pronephros is
fully matured at 4 dpf [58]. Hence, due to the rapid
embryogenesis of the zebrafish, future studies have to employ
different treatment periods to analyze the impact of compounds
at the different stages of nephrogenesis [42]. Table S2. Glomerular alterations in zebrafish larvae
following drug treatment. (DOCX) Table S3. Tubular angle and distance in zebrafish larvae
following drug treatment. (DOCX) Macro S1. Fiji macro to crop images and generate
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CD: 005091. 12. d’Alençon CA, Peña OA, Wittmann C, Gallardo VE, Jones RA et al. (2010) A high-throughput chemically induced inflammation assay in
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21176202. 33. Hentschel DM, Park KM, Cilenti L, Zervos AS, Drummond I et al. (2005) Acute renal failure in zebrafish: a novel system to study a
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Stages of embryonic development of the zebrafish. Dev Dyn 203:
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Automated high-throughput mapping of promoter-enhancer interactions
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Chemical modifier screen identifies HDAC inhibitors as suppressors of
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Developmental toxicity screening in zebrafish. Birth Defects Res C
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intraflagellar transport mutants disrupt cilia but do not affect hedgehog
signaling. Dev Dyn 238: 1744-1759. doi:10.1002/dvdy.21999. PubMed:
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approaches and technologies. Birth Defects Res B Dev Reprod Toxicol
92: 413-420. doi:10.1002/bdrb.20315. PubMed: 21770025. 26. Sukumaran S, Perkins BD (2009) Early defects in photoreceptor outer
segment morphogenesis in zebrafish ift57, ift88 and ift172 Intraflagellar
Transport mutants. Vision Res 49: 479-489. doi:10.1016/j.visres. 2008.12.009. PubMed: 19136023. pp
g
p
92: 413-420. doi:10.1002/bdrb.20315. PubMed: 21770025. 6. Pepperkok R, Ellenberg J (2006) High-throughput fluorescence
microscopy for systems biology. Nat Rev Mol Cell Biol 7: 690-696. doi:
10.1038/nrm1979. PubMed: 16850035. 27. Author Contributions Conceived and designed the experiments: JHW JG SG MS
PLB. Performed the experiments: SS JHW JG MS. Analyzed
the data: JHW SS MS JG. Contributed reagents/materials/
analysis tools: SG. Wrote the manuscript: JHW JG MS. Designed the orientation tool: SG JG. Generated data handling
and visualization tools: JG. Designed morpholino injections
experiments: MS PLB. Designed the study: JHW JG. Finally, for genuine high content screening, an automated
image analysis pipeline for extracted morphological features
would be highly beneficial [22,60]. Moreover, compounds
influencing
kidney
function
without
altering
pronephros
morphology cannot be identified using this pipeline. Thus,
protocols for the large scale analysis of kidney function need to PLOS ONE | www.plosone.org 11 December 2013 | Volume 8 | Issue 12 | e82137 Automated Screening Platform for Zebrafish Kidneys Supervised the study: JHW JG UL. Gave general advice and
edited the manuscript: BT. Supervised the study: JHW JG UL. Gave general advice and
edited the manuscript: BT. Automated Screening Platform for Zebrafish Kidneys Automated Screening Platform for Zebrafish Kidneys U S A 107: 17315-17320. doi:10.1073/pnas.1008209107. PubMed:
20855591. 51. Rosenbaum JL, Witman GB (2002) Intraflagellar transport. Nat Rev Mol
Cell Biol 3: 813-825. doi:10.1038/nrm952. PubMed: 12415299. 41. Neto JA, Oliveira-Filho RM, Simões MJ, Soares JM Jr., Kulay L Jr. (2004) Long-term acetaminophen (paracetamol) treatment causes liver
and kidney ultra-structural changes during rat pregnancy. Clin Exp
Obstet Gynecol 31: 221-224. PubMed: 15491069. 52. Scholey JM (2003) Intraflagellar transport. Annu Rev Cell Dev Biol 19:
423-443. doi:10.1146/annurev.cellbio.19.111401.091318. PubMed:
14570576. 53. Ocbina PJ, Eggenschwiler JT, Moskowitz I, Anderson KV (2011)
Complex interactions between genes controlling trafficking in primary
cilia. Nat Genet 43: 547-553. doi:10.1038/ng.832. PubMed: 21552265. 42. Peng HC, Wang YH, Wen CC, Wang WH, Cheng CC et al. (2010)
Nephrotoxicity assessments of acetaminophen during zebrafish
embryogenesis. Comp Biochem Physiol C Toxicol Pharmacol 151:
480-486. doi:10.1016/j.cbpc.2010.02.004. PubMed: 20170747. 54. de Vries J, Yntema JL, van Die CE, Crama N, Cornelissen EA et al. (2010) Jeune syndrome: description of 13 cases and a proposal for
follow-up
protocol. Eur
J
Pediatr
169:
77-88. doi:10.1007/
s00431-009-0991-3. PubMed: 19430947. 43. Shrim A, Berger H, Kingdom J, Hamoudi A, Shah PS et al. (2005)
Prolonged exposure to angiotensin-converting enzyme inhibitors during
pregnancy. Fetal toxicity could be reversible. Can Fam Physician 51:
1335-1337. PubMed: 16250418. 55. Friedland-Little JM, Hoffmann AD, Ocbina PJ, Peterson MA, Bosman
JD et al. (2011) A novel murine allele of Intraflagellar Transport Protein
172 causes a syndrome including VACTERL-like features with
hydrocephalus. Hum Mol Genet 20: 3725-3737. doi:10.1093/hmg/
ddr241. PubMed: 21653639. 44. Martinovic J, Benachi A, Laurent N, Daikha-Dahmane F, Gubler MC
(2001) Fetal toxic effects and angiotensin-II-receptor antagonists. Lancet 358: 241-242. doi:10.1016/S0140-6736(01)05426-5. PubMed:
11480433. 56. Sun Z, Amsterdam A, Pazour GJ, Cole DG, Miller MS et al. (2004) A
genetic screen in zebrafish identifies cilia genes as a principal cause of
cystic kidney. Development 131: 4085-4093. doi:10.1242/dev.01240. PubMed: 15269167. 45. Pryde PG, Sedman AB, Nugent CE, Barr M Jr. (1993) Angiotensin-
converting enzyme inhibitor fetopathy. J Am Soc Nephrol 3: 1575-1582. PubMed: 8507813 57. Sabaliauskas NA, Foutz CA, Mest JR, Budgeon LR, Sidor AT et al. (2006) High-throughput zebrafish histology. Methods 39: 246-254. doi:
10.1016/j.ymeth.2006.03.001. PubMed: 16870470. 46. Quan A (2006) Fetopathy associated with exposure to angiotensin
converting enzyme inhibitors and angiotensin receptor antagonists. Early Hum Dev 82: 23-28. doi:10.1016/j.earlhumdev.2005.11.001. PubMed: 16427219. 58. References (2012) Mechanism-based urinary biomarkers to identify the potential for
aminoglycoside-induced nephrotoxicity in premature neonates: a proof-
of-concept study. PLOS ONE 7: e43809. doi:10.1371/journal.pone. 0043809. PubMed: 22937100. 14. Kokel D, Bryan J, Laggner C, White R, Cheung CY et al. (2010) Rapid
behavior-based identification of neuroactive small molecules in the
zebrafish. Nat Chem Biol 6: 231-237. doi:10.1038/nchembio.307. PubMed: 20081854. 35. Kent AL, Maxwell LE, Koina ME, Falk MC, Willenborg D et al. (2007)
Renal glomeruli and tubular injury following indomethacin, ibuprofen,
and gentamicin exposure in a neonatal rat model. Pediatr Res 62:
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Glomerular nephrotoxicity of aminoglycosides. Toxicol Appl Pharmacol
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and amikacin-induced nephrotoxicity in patients with normal baseline
renal function. Fundam Clin Pharmacol 23: 515-520. doi:10.1111/j. 1472-8206.2009.00702.x. PubMed: 19709328. 17. Drummond IA (2005) Kidney development and disease in the zebrafish. J Am Soc Nephrol 16: 299-304. doi:10.1681/ASN.2004090754. PubMed: 15647335. 18. Drummond IA, Davidson AJ (2010) Zebrafish kidney development. Methods
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mitochondria, and cell death mechanisms in drug-induced liver injury:
lessons learned from acetaminophen hepatotoxicity. Drug Metab Rev
44: 88-106. doi:10.3109/03602532.2011.602688. PubMed: 22229890. 19. Perner B, Englert C, Bollig F (2007) The Wilms tumor genes wt1a and
wt1b control different steps during formation of the zebrafish
pronephros. Dev Biol 309: 87-96. doi:10.1016/j.ydbio.2007.06.022. PubMed: 17651719. 20. Westerfield M (2000) The zebrafish book. A guide for the laboratory
use of zebrafish (Danio rerio), 4th edition. University of Oregon Press,
Eugene, OR. 40. North TE, Babu IR, Vedder LM, Lord AM, Wishnok JS et al. (2010)
PGE2-regulated wnt signaling and N-acetylcysteine are synergistically
hepatoprotective in zebrafish acetaminophen injury. Proc Natl Acad Sci 12 PLOS ONE | www.plosone.org December 2013 | Volume 8 | Issue 12 | e82137 Automated Screening Platform for Zebrafish Kidneys Automated Screening Platform for Zebrafish Kidneys Kramer-Zucker AG, Wiessner S, Jensen AM, Drummond IA (2005)
Organization of the pronephric filtration apparatus in zebrafish requires
Nephrin, Podocin and the FERM domain protein Mosaic eyes. Dev Biol
285: 316-329. doi:10.1016/j.ydbio.2005.06.038. PubMed: 16102746. 47. Coleman CM, Minor JJ, Burt LE, Thornhill BA, Forbes MS et al. (2007)
Angiotensin AT1-receptor inhibition exacerbates renal injury resulting
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Zebrafish: a model system for the study of vertebrate renal
development, function, and pathophysiology. Curr Opin Nephrol
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20:
416-424. doi:10.1097/MNH.0b013e3283477797. PubMed: 21519251. 48. Friberg P, Sundelin B, Bohman SO, Bobik A, Nilsson H et al. (1994)
Renin-angiotensin system in neonatal rats: induction of a renal
abnormality in response to ACE inhibition or angiotensin II antagonism. Kidney Int 45: 485-492. doi:10.1038/ki.1994.63. PubMed: 8164437. 60. Vogt A, Cholewinski A, Shen X, Nelson SG, Lazo JS et al. (2009)
Automated image-based phenotypic analysis in zebrafish embryos. Dev Dyn 238: 656-663. doi:10.1002/dvdy.21892. PubMed: 19235725. 49. Loria A, Reverte V, Salazar F, Saez F, Llinas MT et al. (2007) Changes
in renal hemodynamics and excretory function induced by a reduction
of ANG II effects during renal development. Am J Physiol Regul Integr
Comp Physiol 293: R695-R700. doi:10.1152/ajpregu.00191.2007. PubMed: 17491111. 61. Rider SA, Tucker CS, del-Pozo J, Rose KN, MacRae CA et al. (2012)
Techniques for the in vivo assessment of cardio-renal function in
zebrafish (Danio rerio) larvae. J Physiol 590: 1803-1809. PubMed:
22331420. 50. Drukker A (2002) The adverse renal effects of prostaglandin-synthesis
inhibition in the fetus and the newborn. Paediatr Child Health 7:
538-543. PubMed: 20046466. PLOS ONE | www.plosone.org PLOS ONE | www.plosone.org 13 December 2013 | Volume 8 | Issue 12 | e82137
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NOTES
ON
MISCELLANEOUS
ENDOPARASITES.
By
T.
Harvey
Jounston,
M.A.,
D.Sc.,
University,
({
Read
before
the
Royal
F.L.8.,
Biology
Queensland,
25th
Dept.,
Brisbane.
Society
of
November,
1918).
CESTODA.
Acanthotaenia
Last
region
year
this
the
intestine
of
gallardt
cestode
a
Johnston.
was
found
infesting
black
snake,
Pseudechts
at
riacus,
killed
addition
to
this
found
just
under
the
peritoneum
company
with
A.
gallardi
there
were
In
individuals
phalus)
by
of
‘
of
sp.
intestine,
Munro
Hull
parasite,
larval
cestodes
a
strongyle,
distributed
while
many
Mr.
reported
from
inalamb
this
from
in
sp.)
body
cavity.
numerous
isolated
(
Kalice-
part
of
the
stomach
harboured
been
previously
had
Queensland.
trigonophora
St.
&
Hass.
1918,
was
found
in
January
It
had
not
been
Eumundi.
(Sparganum
greater
entozoa
host
tapeworm
at
and
No
Moniezia
This
csophagus
porphyIn
Diaphanocephalus
the
mid-
Eumundi.
the
were
throughout
the
Physaloptera.
of
the
in
numbers
previously
reported
Queensland.
Multiceps
Dr.
Dodd
(1918,
p.
parasites
of
live
stock
statement
that
“‘
not
J
have
the
multiceps
502)
in
Leske.
a
recent
address
referred.
to
the
the
condition
Gid
is
not
common
been
able
obtain
any
records
cases
of
occurrence
known
as
Cenurus
cases
)
of
so-called
to
of
parasite
on
and
endo-
made
in
N.S.W.”
of
authentic
worm,
generally
the
gid
bladder
cerebralis,
in
any
part
of
Australia,
gid
traceable
to
other
causes.
being
a
all
210
NOTES
Since
the
Dr.
parasite
information.
ON
MISCELLANEOUS
ENDOPARASITES.
Dodd’s
remarks
seem
he
stated
that
he
had
not
personally
Gid
in
State,
but
had
been
informed.
in
Melbourne.
it
had
been
He
went
on
to
statement
can
be
case
of
say,
state
causal
seen
and
though
proof
parasite
has
all
that
State
he
had
in
this
continent.*
believed
that
gid
were
probably
due
the
list
hydatids
of
for
the
in
the
by
Dr.
Dodd,
cannot
at
given
we
the
It
must
known
first
time.
as
as
occurring
Pall.
the
in
cavity.
affirm
actually
pigs
to
present
entozoa
in
persons.
as.
Australia.
of
In.
well
in
hydatigena
case.
cranial
multiceps
presence
tenuicollis)
noted
for
evidence
occasional
sheep
and
Multiceps
the
of
Gid,
Tenia
now
numbers
collected,
off
is
large
and
view
(Cystieercus
examined
He
conditions
In
The
demonstrated.
that
Hchinococcus
struck
because
possible
of
be
not.
quite
presence
of
N.S.W.,
been
mistaken
occurrence
in
exist
is
was
cases
have
not
he
the
it
the
I
that
before
not
that
of
which
necessary
a
some
the
is
encountered
confused
of
occasionally
having
agreed
other
asking
conclusive,
does
without
had
that
as
Gid
he
letter
there
that
that
later
that
accepted
mentioned
a
further
reply
that
over
for
I
a
the
that:
N.S.W.,
with
to
imply
in
In
wrote
to
occurs
met
prepared
would
bladder
south-eastern
worm
stage
Queensland
*In the same paper (p. 506) Dr. Dodd refers to the presence of
Trichostrongylus axei (Strongylus gracilis) and other stomach worms in
Australian sheep and cattle. (See also Dodd, Ann. Rep. Dept. Agric. Q’land,
1909, p. 93-4, where he calls it Str. gracilis). Cobbold (Parasites, etc., 1879,
p. 283) gave the name Strongylus axei to certain minute nematodes occurring in the mucous membrane of the stomach of donkeys. Railliet and
Henry (C. R. Soe. Biol., 66, 1909, p. 87) placed S. gracilis McFad. and 8.
extenuatus Raill. as synonyms of T'richo. axei Cobbold. Ransom in his
fine account of the nematodes occurring in the alimentary tract of sheep,
cattle, etc. (Bull. 127, U.S. Dept. Agric., B.A.I., 1911, p. 94) referred to.
S. gracilis McF. as a synonym of Railliet’s Tricho, extenuatus, making no
mention of Tr. axe: as a parasite of stock—apparently not accepting
Railliet’s views. Leiper (Jour. London Sch. Trop. Med., 1 (1), 1911, p. 25)
in his check list, mentioned T'r. axe7 as a parasite of Equide and (l.c. 1
(2), 1912, p. 116) queried its presence in cattle. In my census of Queensland endoparasites (1916) I referred to Dodd’s record of Sir. gracilis as
Tr. extenuatus Raill, following Ransom (1911) rather than Railliet (1909).
Tn view of these remarks 7'r. axei cannot yet be added to our known helminth fauna unless it be admitted that the two names Str. axei Cobbold
and S. gracilis McFadyean (not Leuckt ‘are synonymous,
BY
T.
HARVEY
Dipylidium
caninum
the
with
which
to
of
maturity
it
may
be
of
interest
this
cestode
as
well
as
egg-bearing
in
a
puppy
specimens
of
canis,
full
in
(Sydney,
September,
211
some
can
L.
Asevidence
reach
rapidity
JOHNSTON.
mention
the
entozoa
that
I
nematode
only
six
found
Toxascaris
weeks
old
1909).
TREMATODA.
Bird
In
some
of
reference
was
made
under
the
my
broad
submitted
Johnston,
them,
along
Host.
Aig
Numenius
cyanopus
Sydney
others,
as
SEW
of
in
his
Australian
certain
were
J.
They
of
flukes
8.
Birds
presence
of
e.g.
Prof.
are
the
parasites
names:
The
Australian
of
generic
etc.
of
lists
to
stomum,
with
Trematodes.
trematodes
Hchinostomum,
to
birds
Mono-
my
friend,
University,
who
described
paper
the
Trematodes
on
(1916).
follows
mes"
:—
|
2,
|
Locality.
|
Q.
|
Echinostoma J., 1912, 1916,—=
| Himasthla harrisoni S.J.J.
Himantopus leucocephalus | South Australia, Monostomum J., 1910.
1912,—Hematotrephus
adelphus 8. J. J.
Charadrius
dominicus
Sydney
Echinostoma
J.,
1910,
=
|
Acanthoparyphium
spinulosum 8. J. J,
Herodias
timoriensis
Hidsvold,
Q.
|
Hchinosioma
J.,
1912
=
_ Echinoparyphium oxyurum
Ibis
molucca
Chenopsis
Micreca
Gladstone,
Parasite.
:
|
Eidsvold,
Q.
atrata
Victoria
fascinans
Eidsvold,
Dolichopera
This
by
trematode
Macalpine
was
(1891,
p.
S a .J. & Patagifer fraternus
S: JL di.
|
Hchinostoma
J.,
1912
=
| Patagifer acuminatus S. J. J
Monostomum
J.,
1910,
1912
_ = Ayptiasmus magnus 8. J.J.
Q.
Echinostoma
J.,
1912
=
Echinoparyphium
harveyanum 8. J. J.
macalpini
originally
40)
superba
in
Victoria.
(which
named)
in
his
he
and
in
as
hosts,
1918
the
gave
tiger
a
genus
description
snake
described
from
Denisonia
Nicoll.
the
Nicoll
(but
not
named)
copper-headed
snake,
included
Dolichopera
of
Noteckis
the
the
(1914,
species
scutatus
species
p.
343),
mentioning
(Victoria),
the
7
dl
be
NOTES
-copperheaded
unnamed
ON
MISCELLANEOUS
snake,
snake
Denisonia
collected
by
superba
Dr.
J.
Island
(1918a).
Quite
recently
he
parasite
and
published
2
figure
(1918b,
p.
ENDOPARASITES.
(Victoria)
B.
and
Cleland*
referred
on
Flinders
briefly
illustrating
an
to
its
the
anatomy
374).
I
obtained
D.
macalpini
from
the
Denisonia
superba
and
the
black
snake
Pseudechis
poi
phy-
riacus
in
Sydney
district,
recording
its
presence
under
the
name
Hemiurus
parasite
of
in
N.S.W.
I
snakes
seems
Australia.
Echinochasmus
tenuicollis
described
by
cormorant,
have
trematodes
sp.
Eastern
Recently
a
Apoblema)
venomous
distributed
from
(
J.
as
from
of
(1910a,
1911).
This
to
fairly
widely
S.
be
J.
Jnstn.
Johnston,
Phalacrocorazx
identified
collected
S.
csophagus
(1916,
p.
melanoleucus,
belonging
to
this
the
same
host
206)
Tuggerah,
species
species
a
few
near
Long-
reach,
Thompson
River,
by
Miss
M.
J.
Bancroft
(August,
1918).
In
specimens
the
vitellaria
extend
further
of
the
ventral
sucker,
relatively
more
remote
my
forwards,
reaching
while
the
male
and
female
glands
from
the
hinder
end
of
animal,
being
situated
the
original
figure.
ventral
sucker
Some
human
the
than
time
blood
posterior
shown
spp.
ago
reference
made
flukes
being
troops
from
(Johnston,
1916,
p.
was
in
possessed
ional
contain
terminal
spines
in
addition.
Thus
at
present
in
In
1917
urine
returned
to
examination,
ova.
Most
specimens
haematobium
Bilh.)
Bilharzia
(S.
mansoni
both
species
State.
It
may
Sambon)
of
human
is
not
species,
presumably
8.
haematobium
has
become
West
summary
Australia.
in
his
Major
article
on
me
from
for
these
in
by
countries
(S.
origin,
Australia
neighbouring
conditions
African
of
and
certain
South
likelihood
the
Egypt
eggs
this
to
the
into
submitted
to
lateral-spined
was
nearer
introduced
37).
Queensland,
found
in
are
Schistosomum
returning
soldiers
the
edge
but
oceas-
were
present
blood
unlikely
become
fluke
that
under
endemic.
One
on
account
established
in
Cherry
gave
Bilharziosis
are
an_
of
one
its
locality
interesting
(1917).
*Dr. Cleland has informed me that the snake was Notechis seulatus.
BY
Capt.
early
Lawton
clinical
(1917,
stools
a
number
Hospital.
In
addition
in
cases,
some
cols,
HARVEY
recently
published
an
of
the
serious
disease
21).
He
found
of
Australian
features
mansoni
of
T.
p.
to
All
account
the
by
S.
ova
in
caused
lateral-spined
Military
the
faces
were
found,
other
parasites
such
intesttnalts,
Lamblia
men
subsequently
the
contain
soldiers
of
Cairo
to
these
21S:
a
Trichomonas
hominis.
JOHNSTON.
ova,
were
in
as
Hntamoeba
and
Blastocystis
discharged
to-
Australia.
NEMATODA.
Dictyocaulus
Dr.
In
1893
T.
L.
Strongylus
on
Diseases
Stock
micrurus
Mehlis,
in
sheep
occurrence
in
horses
in
Sydney,
using
the:
almost
certainly
D.
arnfields
from
the
Brisbane)
and
their
parasite
(Sydney
(Queensland).
is
and
S.
micrurus
(=D.
and
D.
viiparus)
lung
worms,
horses.
Perrie
filaria
from
sheep-
is
lung
worm
goat’s
lung
the
calves.
Synthetocaulus
Dr.
Cleland
parasitised
by
nematodes
identified
as
S.
extremely
brief
account
of
the
had
not
been
me.
J.
capillarts
B.
in
forwarded
goats
in
collected
a
dozen
by
have
braziliense
specimens
rat,
kindly
man,
Director
of
the
Queensland
rattus.
This
the
only
occasion
reported
from
Z£.
H.
p. 96).
From
fowl
though
in
the
to
to
reported
as.
retusa
occur
the
in
were
from
Mr.
H.
Long-
Museum,
as
by
which
I
Brisbane
A.
H.
Hpimys
braziliense
have
recorded
and
Sydney
its.
(1918,
Raill.
alimentary
previously
strongyle
1918)
on
rattus.
Capillaria
Brisbane—not
known
available
May,
determined
in
of
tiny
(Brisbane,
norvegicus
walls
an
Travassos.
this
a
in
Taylor
of
of
presence
Only
is
previously
B.
been
provisionally
District).
nematode
B.
is
a
been
(Sydney
Miss
intestine
of
Australia.
Heligmosomum
About
Muller.
portion
which
capillaris
Lungworms
occurring
has
of
Queensland,
occurrence
name.
the
in
the
same
of
report
recorded
to
horse
his
Cobbold.
Bancroft
referred
The
in
arnfieldt
canal
recorded
Sydney
and
of
from
the
domestic
Queensland.
Melbourne.
214
NOTES
ON
MISCELLANEOUS
Pneumonema
This
Tiliqua
the
tilique
was
collected
recently
scincoides
on
Facing
Island
recorded
having
been
vegicus.
In
(Port
tetraptera
from
found
in
Queensland
Curtis)
and
in
Mus
first
time,
and
in
Epimys
nor-
musculus
only
one
worm,
collected.
This
constitutes
the
first
the
nematode
in
(1916)
mentioned
by
Dr.
Hall
(collected
by
Miss
B.
agrees
in
all
given
by
Hall.
particulars
the
indebted
to
a
a
rat
Taylor,
Sept.,
with
the
account
nasuta
G.
female,
of
the
is
not
which
host.
B.
Miss
mature
record
brown
as
Cheilospirura
am
of
the
case
I
lungs
for
latter
of
the
Nitzsch.
the
presence
of
from
District.
Oxyuris
Now
~
Johnston.
worm
Brisbane
was
ENDOPARASITES.
The
specimen
1918,
Brisbane)
of
the
species
Rud.
Y.
James,
B.Sc.
for
specimens
a
domestic
this
worm
collected
from
the
urodeum
of
fowl
at
West
Burleigh,
1918—not
previously
recorded
this
State.
Habronema
The
occurrerice
Australian
horses
of
musce
the
has
and
history
has
been
The
fully
followed
He
did
Ransom
(1913).
recently
published
an
have
tumours
Habronema
granulomata
of
horses
to
which
of
three
species
horse
these
re-examined
my
have
microstoma
appears
to
Sydney
the
larve
belonged.
material
labelled
found
be
the
that
many
years
ago
by
Mr.
A.
S.
Le
Soeuf,
Habronema
was
collected
numbers.
Spiroptera
musce
Specimens
small
**
in
H.
in
Habronema
form
Brisbane,
on
the
of
although
commoner
in
habronemiasis.”’
as
and
present
Australia
cancer”
“‘
the
horses,
South
of
care-
in
and
form
Queens-
of
Victoria
another
and
in
in
account
be
microstoma’’
collected
interesting
or
“‘“swamp
stage
to
know
the
I
certain
believed
not
infesting
life
by
Territory
been
N.S.W.
1912).
He
has
in
(Johnston,
(1916).
its
flies
land
larval
though
certain
in
by
in
larval
present
recorded,
musce
the
be
caused
H.
since
to
of
of
inferred,
reported
occurrence
stage
been
be
Northern
adult
yet
could
Bull
Carter.
not
presence
from
in
material
also
Melbourne
re-examination
were
all
found
recorded
by
H.
BY
T.
to
belong
must
be
and
occurring
in
N.S.W.
Dr.
J.
B.
the
in
nodules
as
of
evidently
musce
nor
H.
added
to
H.
in
from
(Sydney
District).
fied
as
recorded
215
as
previously
known
entozoa
*
cavities
in
dense
horses
in
West
refer
to
H.
has,
as
of
fibrous
Australia.
as
I
tissue
in
of
his
Neither
4H.
been
noted
Both
megastomat.
far
Spiroptera
know,
State.
Dr.
the
of
presence
Gongylonema
tumours
list
the
or
that
year,
the
recorded
microstoma
occurring
Last
microstoma
Queensland.
Cleland}
stomach
records
to
JOHNSTON.
me.
musce
microstoma
HARVEY
ingluvicola
Cleland
forwarded
some
small
crop
gizzard
of
have
been
walls
of
The
causal
G.
ingluvicola,
which,
as
far
as
only
from
locality
in
the
United
a
the
Ransom.
and
parasites
I
worm
a
know,
chicken
indenti-
has
States
been
(Ransom,
1904).
ACANTHOCEPHALA.
*
This
Centrorhynchus
asturinus
parasite,
infests
novaehollandiae
as
a
Gm.,
Gigantorhynchus
Centrorhynchus,
the
summarised
by
three
long
tubular
ones
as
I
which
shown
have
specimens
hawk,
Baza
gum,
near
but
is
my
the
now
Cleave
as
of
glands
The
to
the
genus
have
been
has
only
male
several
smaller
(1913,
pl.
17,
41).
figure
to
the
J.
B.
subcristata
Gd.,
in
December,
well
described
of
belonging
as
Astur
instead
Dr.
N.S.W.,
was
which
by
Casino,
hawk,
transferred
(1916).
original
white
Queensland,
characters
cement
identified
collected
North
chief
Van
in
in
Johnston.
as
Cleland
a
fig.
species
from
a
a
1916
at
number
in
few
crested
Mummul-
the
collec-
*My attention has recently been drawn to a reference in the Scientific
Australian (June 1918), where it stated that a paper by Mr. G. F. Hill,
dealing with the life histories of Habrone na musce, +4. microstoma and H.
megastoma, had been read before the May meeting (1918), of the Heyal
Society of Victoria.
TCleland. Diseases of Animals, etc., Bull 33, Dept. Agric.
Australia, 1909, p. 3; Trypanosomiasis, etc. Bull. 34, 1909, p. 15.
West
{Desmond (Journ. Agric. Ind. South Australia, 7, 1904, p. 569) referred
to the presence of Sclerostomum hypostomum in tumours in the stomach
of South Australian horses. The reference should be H. megastoma.
216
NOTES
ON
tion
of
ville,
obtained
in
North
Astur
clarus
(A.
cinereus
A.
the
MISCELLANEOUS
Australian
Brienl,
the
opportunity
to
The
found
Institute
in
of
boobook
District—T.H.J.,
rhynchus
sp.
riacus
presence
in
my
specimens
already
referred
district.
It
intestine
though
specimens,
for
this
is
as
been
Bancroft
also
a
typical
to
having
as
frequents
the
generally
more
been
in
the.
Centro-
Pseudechis
poi
phy-
the
first
from
the
black
snake
in
the
Eumundi
killed
three
time
quarters
abundant
hirundinaceus
the
for
been
lower
me
Johnston.
in
taken
Dr.
having
Dr.
recorded
having
thank
by
parasite
now
Goshawk,
giving
rotundocapitatus
of
Hormorhynchus
to
mentioned
is
Towns—
gray
desire
his
1912)
Queensland
the
Institute,
(collected
Echinorhynchus
The
I
was
Medicine,
from
the
which
Kidsvold
Tropical
Vieill).
examine
Ninox
of
Queensland
dircctor
echinorhynch
ENDOPARASITES.
in
of
;
the
the
rectum.
(Gigantorhynchus
gigas
Goze).
This
is
met
Queensiand—not
with
occasionally
previously
in
reported
pigs
in
from
south-eastern.
this
State.
*
LINGUATULIDA.
Porocephalus
A
black
snake
teretiusculus
Pseudechis
porphyriacus
—referred
to
earlier
in
sitised
the
above
pentastome,
the
lung,
while
the
small
males
organ.
This
constitutes
in
by
the
extremity
of
of
the
now
the
presence
known
Pseudechis
and
tiger
snake
known
Queensland
to
D.
infest
the
Notechis
and
I
to
the
islands
larval
denticulatum,
is
stage,
now
the
Australian
(Blue
of
Strait.
record.
which
is.
snakes—
from
Mountains,
Australia
the
Diemenia.
taken
Western
generally
at
superba,
specimens
para-
distributed.
first
following
Bass
be
found
entozoon,
serrata
definitely
being
were
Kumundi
to
the
have
from
Tinguatula
The
females
of
scutatus
extends
from
found
Denisonia
reticulata.
range
paper—was
Queensland
porphyriacus,
textilis
Its
in
this
Baird.
to
the
N.S.W.).
Southerr-
¥rol.
known
recorded
as
as
Pentastomum.
oceurring
in
the
mesenteric
glands
occasionally
found
BY
T.
of
cattle
in
HARVEY
in
JOHNSTON.
2G
specimens
being
Queensland,
animals
killed
in
the
Brisbane
the
Brisbane
district
abattoirs.
ACARIDA.
Cytodites
This
in
the
mite
is
found
sacs
of
the
air
mange
opportunity
to
(Sarcoptes
muris,
domestic
on
Epimys
Sydney
by
I
rattus
on
‘Dr:
record
rattus
J.-B:
Cleland.*)
have
already
in
N.S.W.
and
Z£.
The
parasite
sets
up
sometimes
the
other
;
the
presence
Raill.)
true
in
norvegicus
and
referred
in
to
a
warty
of
Knemidocoptes
mutans
domestic
in
of
I
take
Notedres
Brisbane
and
the
muris
Melbourne
in
Launceston
Adelaide
on
“rats’’
(collected
occurrence
in
#.
norvegicus
in
Perth,
West
Australia.
of
the
tail,
ears
causes
~~
scaly-leg’’ bE in
its
condition
the
entozoa,
;
alexandrinus
parts
fowl
Miégnin.
not
E.
#.
fowl.
are
alepis
and
in
muris
mites
S.
Viz.
occasionally
Notedres
Though
nudus
and
and
head.
Rob.
Brisbane.
BIBLIOGRAPHY.
1916 Bull, L. B. A granulomatous affliction of the horse, Habronemie
granulomata. Journ. Comp. Path. Therap., 29, (3). (Reprint
15pp.)
1917 Cherry, T., Bilharziosis, ete. Pamphlet, Defence Dept. Commonwealth of Australia.
1918 Dodd, S. Some observations on the internal parasites of live
stock. Agr. Gaz. N.S.W., 1918, pp. 497-507.
1916
Hall, M. Nematode parasites
Mus., 50, pp. 1-258.
of
Rodents,
etc.
Proc.
U.S.
Nat.
1910 a Johnston, T. H. (Notes and Exhibits) P.R.S. N.S.W., 44, p. xviii.
6b — —— On Australian Avian Entozoa. P.R.S. N.S.W., 44,
pp. 84-122.
911 ————— Census of Australian Entozoa. P.R.S.Q., 23, pp. 233249.
“Dr Cleland has recently referred to the presence of the parasite in
Sydney rats. P.R.S. N.S.W 1918, p. 110.
218
NOTES
ON
MISCELLANEOUS
ENDOPARASITES,
1912 a ————— The internal parasites recorded from Australian Birds,
“Emu,” Oct., 1912, pp. 105-112.
1912 ) —————_ Notes on some Entozoa. P.R.S.Q., 24, pp. 63-91.
1913
———— Report on Cestoda and Acanthocephala.
Inst. Trop. Med., 1911 (1913), pp. 75-06.
Rept.
Aust.
1916 ———— Census of the endoparasites recorded as oceurring in
Queensland, etc. P.R.S.Q., 28, pp. 31-79.
1918 ————— Notes on certain entozoa of rats and mice, etc., P.R.S.Q.
30 (3), pp. 53-78.
1916
Johnston,
8.
J.
Ongthe
Trematodes
of
Australian
Birds,
P.R.S.,
N.S.W., 50, pp. 187-261.
1917 Lawton, F. B. Schistosomum mansoni. Early clinical features of
the disease. In pamphlet ‘‘ Bilharziosis.”” See under T. Cherry.
1891 Macalpine, D. Remarks on a fluke parasite in the copperhead
snake.
P.R.S.
Vict.,
3,
pp.
40-45.
1914 Nicoll, W. Trematode parasites of North Queensland. 1. Parasitology, 6 (4), pp. 333-350.
1918 a —————
290-3.
Dolichopera macalpimi, etc. Parasitol., 10 (2), pp.
1918 6 —_——— The trematode parasites of North Queensland, No. iv.
l.c., 10 (3), pp. 368-374.
1904 Ransom B. A new nematode (Gongylonema ingluvicola) parasitic
in the crop of chickens. U.S.D.A., B.A.L., Cire. No. 64, 3 pp.
1913 -————— [he life history of Habronema musce, ete. U.S.D.A.,
B.A.1., Bull. 163, 36 pp.
1916 Van Cleave, H. Acanthocephala of the genera Centrorhynchus and
Mediorhynchus, ete. Trans. Amer. Micr. Soc., 35, pp. 221-232.
Johnston, T. Harvey. 1918. "Notes on miscellaneous endoparasites." The
Proceedings of the Royal Society of Queensland 30, 209–218.
https://doi.org/10.5962/p.91011.
View This Item Online: https://www.biodiversitylibrary.org/item/49266
DOI: https://doi.org/10.5962/p.91011
Permalink: https://www.biodiversitylibrary.org/partpdf/91011
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| 24,818
|
https://arxiv.org/abs/2111.01239
|
arXiv
|
Open Science
|
CC-By
| 2,021
|
Refundable income annuities: Feasibility of money-back guarantees
|
Moshe A. Milevsky and Thomas S. Salisbury
|
English
|
Spoken
| 15,296
| 33,606
|
Refundable Income Annuities:
Feasibility of Money-Back Guarantees
Moshe A. Milevsky and Thomas S. Salisbury
(Date: Version: 26 August 2021)
###### Abstract.
[Short version:] Refundable income annuities (IA), such as cash-refund and
instalment-refund, differ in material ways from the life-only version beloved
by economists. In addition to lifetime income they guarantee the annuitant or
beneficiary will receive their money back albeit slowly over time. We document
that refundable IAs now represent the majority of sales in the U.S., yet they
are mostly ignored by insurance and pension economists. And, although their
pricing, duration, and money’s-worth-ratio is complicated by recursivity which
will be explained, we offer a path forward to make refundable IAs tractable.
A key result concerns the market price of cash-refund IAs, when the actuarial
present value is grossed-up by an insurance loading. We prove that price is
counterintuitively no longer a declining function of age and older buyers
might pay more than younger ones. Moreover, there exists a threshold valuation
rate below which no price is viable. This may also explain why inflation-
adjusted IAs have all but disappeared.
Milevsky is a Professor of Finance at the Schulich School of Business, York
University, and Executive Director of the IFID Centre. Salisbury is a
Professor in the Department of Mathematics and Statistics at York University.
The authors acknowledge funding from the IFID Centre (Milevsky) and from NSERC
(Salisbury) as well as helpful comments from Narat Charupat, Branislav
Nikolic, and participants at the (virtual) IME Congress in early July 2021.
The corresponding author (Salisbury) can be reached at: salt@yorku.ca
Title: Refundable Income Annuities:
Feasibility of Money-Back Guarantees
Authors: Moshe A. Milevsky and Thomas S. Salisbury
Abstract [Long version]:
This paper offers some basic theorems and observations related to: (i) cash
refund and (ii.) instalment refund income annuities (IAs). We distinguish
between the price paid by annuitants and the actuarial present value of the
future payments received by the annuitant. These differ when a loading is
applied, but are obviously the same in the unloaded case. We start by proving
that unloaded prices are a declining function of issue age $x$ and valuation
rates $r$, neither of which are trivial statements when dealing with
refundable products. We also compute measures of annuity duration and show how
they differ from traditional interest rate sensitivity, again due to the
refund feature.
In terms of contribution, our main (novel) result relates to loaded cash-
refund IAs, that is once the value is grossed-up by $(1+\pi)$ and converted to
a market price paid by or charged to annuitants. Under mild assumptions the
price of a cash-refund IA is no longer a declining function of age and older
annuitants might have to pay more than younger ones for the same lifetime of
income. More importantly, there exists a threshold valuation rate $r_{\pi,x}$,
at which the pricing function of a cash-refund IA is no longer viable.
Practically speaking, if-and-when the insurance company or pension fund
assumed investment return (i.e. portfolio yield) is projected to fall under
$r_{\pi,x}$, they are no longer able to offer refundable IAs at any price.
Finally, our paper might help explain why inflation-adjusted income annuities
have all-but disappeared from the retirement landscape. Real interest rates
are currently far under our $r_{\pi,x}$, which means that real refunds are no
longer feasible.
JEL: G12 (Asset Pricing), G22 (Actuarial Studies), G52 (Insurance)
Keywords: Pensions, Annuities, Retirement Planning, Bond Duration.
## 1\. Introduction and Motivation
Per the SECURE Act of 2019, also known as the Setting Every Community Up for
Retirement Enhancement Act, the U.S. Department of Labor has been tasked with
creating a set of technical regulations and mathematical assumptions for
reporting the account balance of all Defined Contribution (DC) retirement
plans in the U.S. as an income stream, together with the usual mark-to-market
value of all securities held at the end of the quarter111See, for example,
https://www.groom.com/resources/lifetime-income-provisions-under-the-secure-
act/.
This regulation comes into effect in late September 2021 and will affect 76.8
million Americans with an ERISA-regulated DC account. As of the effective date
and assuming it goes thru as planned every single plan sponsor and
administrator will be mandated to calculate how much life annuity income the
balance would buy, if the participant decided to convert the lump sum into a
lifetime of income. This applies to participants at all ages and not just at
or near retirement.
Thus for example, a 40-year-old employee working at a meat packaging plant in
Nebraska will be informed on their quarterly 401(k) statement that the
$100,000 balance in their 401(k) plan would be equivalent to (only) $5,000 of
lifetime income because the actuarial pricing factor – the denominator by
which the $100,000 account balance is divided – is 20.
The policy objective of this entire exercise – which the Department of Labor
is estimating will cost employers $200 million to implement in just the first
year – is to nudge participants to anchor their limited attention on a
standard of living instead a lump-sum of money222See
https://www.govinfo.gov/app/details/FR-2020-09-18/2020-17476 for the Interim
Final Rules issued by the Department of Labor and printed in the Federal
Register. This is a laudable goal, familiar to financial economists as the
foundation of the life-cycle model.
To be clear, the calculation by which the ongoing balance is converted into a
lifetime income will be based on a theoretical formula for the value of life
annuity and the math itself has been the subject of much controversy and
debate over the last year. While insurance companies that hope to sell
annuities to many of those 76.8 million participants welcome the spotlight on
life annuities, traditional asset managers who offer their own decumulation
solutions aren’t as welcoming333See for example
https://www.ft.com/content/cbbd43cb-e80c-4e81-8fb2-dfced5f7b62e.
To this end, the Department of Labor has released (and then updated) a 30-page
document with proposals for the discount rates, the mortality rates, the
annuity riders and even the magnitude of loadings, after receiving public
comments about these matters. In other words, there is an enormous amount of
interest among a vast array of businesses about how to value a stream of
lifetime income. Indeed, a topic that was of interest to a small number of
specialized insurance actuaries has gone mainstream. Quite ironically and as
most pension economists are well-aware, few if any participants in DC plans
voluntarily purchase life annuities with their account balances, which is yet
another interesting aspect of the proposed regulation. Will it help?
Indeed, the literature has long-known and puzzled over the thin market for
voluntary annuities and the fact few people actively annuitize their wealth at
retirement444See Modigliani (1988). Annuitization is the process of exchanging
a lump-sum of money for a stream of lifetime income that can’t be outlived,
via products sold by insurance companies but also offered by traditional
Defined Benefit (DB) pension funds. The formal literature began with Yaari
(1965) who proved that, absent bequest motives, a utility-maximizing consumer
should annuitize all wealth. And ever since Yaari (1965), many explanations
have been proposed for why most people don’t actually do this, and just as
many remedies have been offered to fix the annuity market, such as mandating
income projections on DC account statements555See Benartzi et al. (2011)..
We should note that the academic annuity literature is mostly about income
annuities (IAs) which are different from tax-sheltered accumulation annuities
intended as long-term savings vehicles. And, although the original paper Yaari
(1965) – and thousands of models and papers that have followed in its path –
deal with actuarial notes (or instantaneous tontines), most economist treat
IAs as the nearest real-world substitute, and the above-noted proposals are
about IAs.
What is less known among economists – and the main impetus for our paper – is
that of the $8-$10 billion of annual premium flowing into IAs in the U.S. in
recent years, the majority is allocated to products with a money-back
guarantee, which of course reduces the embedded longevity insurance, risk
pooling and mortality credits. According to data from CANNEX Financial
Exchanges666CANNEX Financial Exchanges is a data vendor that provides annuity
quotes and conducts a quarterly survey of the types of annuities sold. Note
that both authors have a financial relationship with CANNEX., the most popular
type of IA quoted777Actual sales are expected to be even more skewed than
quotes, since duplicate quotes are more likely to be purchasers of refundable
IAs assessing the guarantee cost. See Charupat et al. (2016) for more details
concerning this dataset. is one in which the annuitant or his/her beneficiary
is guaranteed their money back.
As noted in Table 1 of this paper888All tables and figures are placed at the
end of the paper., in general there are three structurally distinct types of
IAs. The first class is a life only product under which death terminates the
income flow. Again, those are closest to the actuarial notes of Yaari (1965).
The second class is an IA in which payments are guaranteed for a fixed number
of years, to a beneficiary if the annuitant is no longer alive. The third is
refundable IAs in which the annuitant has a guarantee that their entire
premium will be returned, either in a lump-sum of cash or instalments. Notice
the trend in Table 1 over time during the last 10 years. In 2011 life only
(a.k.a. Yaari) IAs were more popular than refundable IAs. But by early 2021
the majority of price quotes are in cash or instalment refund products.
Table 1 Placed Here
Note that our focus here – and the data displayed in the table – is on single-
life annuities. But joint-life annuities, which we do not treat in this work,
also display the same pattern over time, towards cash-refund and instalment
refund features. Naturally the demand and need for a money back guarantee is
reduced because it’s only after the second death that the annuity payment is
extinguished. Either way, we leave the analysis of two (or more) lives to
other authors and papers.
To be clear, here is a simple example that should explain the mechanics of a
cash versus instalment IA (without loading) and by extension the recursivity
problem which is the focus of this paper. Assume an IA price that is: $a=12.5$
per $1 of lifetime income. This means that if the annuitant deposits or
provides a premium of $P=\$100,000$, it leads to an income of $\$C=P/a=$ 8,000
per year for life. Now, if this particular IA is a life only product, then
death extinguishes the contract and the beneficiaries aren’t entitled to any
residual value. But, if this is a cash-refund IA, and if the annuitant happens
to die after exactly 10 years of payments the estate or beneficiary receives a
lump-sum cash payout of: $(\$100,\\!000-\$80,\\!000)=\$20,\\!000$. But, if
death occurs after year 12.5, the estate receives nothing.
So, with a cash-refund IA, the premium $P$ generates a lifetime cash-flow $C$,
plus a death benefit $(P-TC)_{+}$, where $T$ is the stochastic date of death.
We now arrive at the main problem and motivation for this paper. Note that the
$P$ appears on both sides of the valuation equation, hence the recursivity.
Likewise, if the IA is of the installment refund type, then if at the time of
death $P<TC$, the annual payments of $C$ continue until time $T=P/C$, by which
time the entire premium is returned. After time $T=P/C$, which in the above
case would be $T=12.5$ years, all payments cease. One again, this induces a
recursivity problem, all of which will be addressed.
One additional motivation for this paper (in mid 2021) is that the consumer
trend towards refundable income annuities, is on a collision course with low
(and possibly negative) interest rates. At the time of writing, the yields on
long-term European government bonds are (mostly) negative999Source: European
Central Bank & IMF Report, December 2020. For example, in late June 2021 the
10-year rate on Greek government bonds fell below zero for the first time..
So, it’s not entirely inconceivable that negative valuation rates will
eventually be used to price and sell IAs in the U.S.. But even if that
scenario is farfetched, these two opposing trends raise a critical question.
Is it viable to sell refundable IAs in a low enough interest rate environment?
Our answer and key result is that there exists a positive lower bound to the
underlying rate, below which cash-refund IAs can no longer be offered for
sale. The type of income annuities that consumers actually want to purchase
simply can’t exist.
### 1.1. Outline of the paper, and key results
The remainder of this paper is organized as follows. In the Section 2 we
position our paper within the broader annuity literature, with particular link
to actuarial science, pension finance and insurance economics. Then, in
Section 3 we introduce and formally present the fundamental valuation
relationship or algorithm for cash refund (hereafter CR) and instalment refund
(hereafter IR) income annuities. We begin that section with basic life only
(hereafter LO) income annuities to set notation and terminology and then
present a variety of calibrated numerical values for LOIA, IRIA and CRIA
assuming Gompertz mortality values.
We should make very clear at this early juncture that our entire paper is
predicated on (i.) deterministic mortality hazard rate, and (ii.) the law of
large number which underlies the actuarial pricing of mortality and longevity
contingent claims. In other words, we assume the insurance company that
manufactures the IAs sells a sufficiently “large” number of policies to
eliminate all idiosyncratic mortality risk and that there is no systematic
mortality risk to worry about. Follow-up research will deal with “small”
portfolios, stochastic mortality hazard rate and the ruin probabilities
associated with refundable IAs.
After that, Section 4 examines the sensitivity of loaded and unloaded consumer
prices to both mortality rates and interest rates, which leads to durations
and convexity. Once again (counterintuitively) as a result of the
recursiveness, we distinguish between a classical measure of Macaulay
duration, that is the time-weighted average of discounted cash-flows, versus a
(log) derivative with respect to valuation rates. In the case of refundable
IAs these two measures differ from each other, echoing the concept of option-
adjusted duration. Next, section 4 also explores the money’s worth ratio of
these products.
Having set the stage with numerical tables and figures, Section 5 contains the
precise statements of our general theorems and formulae, along with their
proofs. Section 6 concludes the paper proper. The appendix Section 7, contains
a brief R-script that can be used to value cash-refund income annuities using
a simple bisection algorithm which rapidly converges on the number that
equates the actuarial present value to the price.
Before we move-on to the section containing the literature review, we briefly
summarize the main theoretical – and what we believe to be are the novel –
results offered in this paper, albeit much less formally:
* •
As long as valuation discount rates denoted by $r$ are positive, there is
always a unique price $a^{\star}$ at which an unloaded CRIA is viable.
Naturally, if $r$ is negative, the buyer can never guarantee their money back,
although a basic life only IA is always viable. When $r>0$ the price of the
CRIA is decreasing in $r$, and decreasing in the age $x$ of issue, assuming
mortality hazard rates increase with age.
* •
But, if $r$ exceeds a certain level, there is a unique price
$\widehat{a}^{\star}$ at which the CRIA with a given loading is viable. Below
a threshold level of $r$, the loaded CRIA fails to be viable. Once again,
assuming that mortality hazard rates rise with age, the loaded CRIA is viable
for $x$ below a certain level, but not for larger ages. That number is in the
mid-70s or early 80s, given current interest rates and typical insurance
loadings.
* •
Regardless of $r$ and the insurance loading, there is a unique price
$a^{\circ}$ or $\widehat{a}^{\circ}$ at which an instalment refund IA is
viable. It is decreasing in $r$, once again assuming an increasing hazard
rate, and declining in age $x$. Typically this price $\to 0$ at advanced ages
in the unloaded case, but not under loading.
## 2\. Position within the literature
This paper is interdisciplinary in nature and overlaps with three distinct
research areas of expertise: (i.) actuarial science, (ii.) pension finance and
(iii.) insurance economics. In this section we review how our paper engages
with the scholarly literature in those three fields.
### 2.1. Actuarial Science
Within the literature on actuarial science, the valuation and pricing of
income annuities is a topic with three centuries of history, starting with the
original paper by Edmund Halley published in Halley (1693), in which he
introduced the idea of discounting for both mortality and interest and applied
it the valuation of income annuities on multiple lives. In fact, by the late
18th century and certainly by the mid 19th century, the valuation and pricing
of income annuities under a given (1.) mortality table and (2.) interest rate
curve, was viewed as fully settled and secure by insurance actuaries101010See
the collection of historical articles contained in Haberman and Sibbett
(1995).. Insurance companies as well as local and national governments issued
immediate, deferred and reversionary (triggered by death) annuities, all well
represented in Victorian novels and literature111111See Poterba (1997, 2014)
as well as the popular article by Merton (2014).. The actuarial curriculum for
valuing income annuities has only seen minor changes during the last half-
century.
However, most if-not-all income annuities sold by private insurance companies
and voluntarily purchased by individual consumers during the last three
centuries did not offer the money-back or cash refund feature that has become
exceedingly popular in the last decade, and is the focus on this paper. Recall
from Table 1 that a cash-refund annuity has become the most dominant rider
added to income annuities. So, while historically one could always purchase an
income annuity with a guarantee period of 5, 20, 15 or 25 year’s period
certain, those had no meaningful impact on the valuation and pricing
methodology. Mathematically, the period certain simply modifies survival
probability from an integrand that is less than 1, to a constant equal to 1,
during the period of income certainty. And, even if the survival curve used
for pricing depends on the guarantee period selected, perhaps due to anti-
selection, this can easily be incorporated into the basic valuation equation
by valuing a deferred income annuity together with a term certain
annuity121212See Finkelstein and Poterba (2004), where they find selection
effects based on guarantee periods..
In contrast to these rather trivial integrals, the cash-refund income annuity
generates a non-trivial and recursive relationship which embeds the initial
value and price into the payout function. In fact as we noted earlier and will
carefully demonstrate in Section 5, this money-back guarantee not-only creates
mathematical complexity, it might actually lead to no feasible solution. There
might not be a fixed point to the valuation equation, or to put it bluntly the
product will not exist if insurance loadings (profits, commissions, expenses,
or risk capital) increase beyond a certain upper threshold, or valuation rates
decline beyond a lower bound.
Now, the classic actuarial textbooks are well-aware of the above-noted problem
or difficulty with valuing the cash-refund feature of income annuities, and do
make reference to an iterative procedure or scheme that is required to solve
for the price of the longevity-contingent claim. But, that discussion is often
relegated to an obscure appendix or perhaps a page or two of extra problems,
whereas entire chapters are dedicated to the valuation of conventional income
annuities131313For example, the second edition of the 800 page actuarial
textbook by Bowers et al. (1997), dedicates a page to cash refund annuities,
as an exercise on page 536. The 480 page textbook by Dickson et al. (2009),
mentions them briefly as an exercise on page 139, and the 370 page textbook by
Promislow (2006) has a page or two devoted to them, abstractly. Needless to
say, these textbooks use traditional actuarial notation, symbols and
expression that are likely to be inaccessible to economists interested in
valuing income annuities. The book by Cannon and Tonks (2008) mention value-
protected annuities, a form of CRIA, but they don’t mention the valuation
problems. More importantly, and as far as the academic contribution of this
paper is concerned, we are unaware of any actuarial textbook or prior article
that considers the conditions under which the loaded cash-refund income
annuity price might not exist.
We suspect the reason for the rather limited (historical) attention to the
cash refund feature is that they simply weren’t popular, and perhaps were even
unavailable. It is our understanding that cash-refunds came into existence
around the time of the introduction of higher estate taxes (in the US, in the
1940s and 1950s), where beneficiaries preferred a lump-sum payment at death to
cover tax liabilities for illiquid assets. We have been told that (old)
annuity company accounting systems were originally designed for ongoing and
periodic cash-flows, not lump-sum death benefits, which created further
impediments to offering cash-refund income annuities141414Source: Author
conversations with insurance executives, and in particular Gary Mettler..
Needless to say, we don’t want to digress into the business history of the
income annuity market in the US, but simply note the above background to
provide context for why actuarial science might have neglected the cash-refund
income annuity as part of its foundational pedagogy. Our paper fills that gap.
To be clear, there are recent papers in the actuarial and insurance literature
that examine the valuation, hedging and risk management of guarantees embedded
within modern 21st century variable annuities with guaranteed living benefits
(GLBs), which are akin to financial put options struck on life or
death151515See for example Xu et al. (2018), Huang et al. (2017b), Moenig and
Bauer (2016), Steinorth and Mitchell (2015), Ngai and Sherris (2011), Milevsky
and Salisbury (2006), and as well Bauer et al. (2008) which examines the many
dimensions of variable annuity guarantees.. The variable annuity market is a
multi-billion dollar industry in the US, and many orders of magnitude larger
than the market for income annuities. But, as their name suggests, those
papers and authors are focused on purely variable products in which separate-
account investment returns fluctuate with markets and interest rates. The
nature of those guarantees and the risks associated with those derivatives are
quite different. And, while those products generate their own mathematical
complexities – and in some circumstances might also fail to be viable – the
focus of our paper once again is on the income annuity at the core of
retirement planning. That is the annuity Edmund Halley, Richard Price,
Benjamin Gompertz (1825), and Jane Austen would recognize.
In sum, for researchers interested in actuarial and insurance pricing, we
contribute to the literature by showing how a simple cash-refund on an income
annuity can lead to challenging and interesting mathematical problems.
Practically speaking we can help explain why inflation-adjusted cash refund
annuities no longer exist; real rates are simply too low.
### 2.2. Pension Finance
The second of the three strands of literature with which this paper engages,
has to do with the so-called money’s worth ratio (MWR) of income annuities,
and the competitiveness and efficiency of the retail income annuity market. In
the early 1990s, a number of well-known pension economists in the U.S.
introduced the MWR in series of books and papers161616See Friedman and
Warshawsky (1990), as well as the article by Mitchell et al. (1999) and the
subsequent book cited as Brown et al. (2001).. They suggested using the
relative ratio of market annuity prices to theoretical annuity model values
(i.e. the MWR) as a measure of price efficiency as well as a way of detecting
anti-selection by healthier annuitants171717See Finkelstein and Poterba (2014)
as well as the earlier noted Finkelstein and Poterba (2004).. The early
research indicated that this ratio or number was between 80% and 90%,
depending on the mortality tables and term structure of interest rate used to
compute theoretical model prices. And, when annuitant (versus population)
mortality and risk-free rates were used to compute the MWR, the ratio often
exceeded 100%. This was taken as a sign the market was efficient and
functioning properly. Recent research work updating the original estimates
continue to point to MWRs in the same range, despite the decline in both
interest rates and mortality rates during the last three decades181818Recent
papers using the MWR are Poterba and Solomon (2021), Blanchett et al. (2021)
as well as Wettstein et al. (2021)..
The economic impetus and relevance of these MWR studies revolve around the
global demise of traditional Defined Benefit (DB) pensions and the trend
towards individual and portable Defined Contribution (DC) investment accounts.
Recall that the default option in a DB plan is retirement income for life,
which obviously resembles the payout of an income annuity. In many cases, it
not only is a default option in the pension plan, it’s the only option
available at retirement. For example, the income annuity provided by U.S.
Social Security (or the Canadian version, CPP) cannot be taken as a lump sum
in cash. In contrast, the default option in a DC plan is either a lump-sum of
money at retirement or perhaps a systematic withdrawal plan with a default
spending rate. Neither of those options protects the retiree against longevity
risk, i.e. the risk of outliving resources. That is the main reason why income
annuities have received such attention from economists over the last few
decades. From a retirement policy perspective, income annuities are being
promoted by both academic economists as well as insurance company executives
to help replace the lost longevity insurance in DC plans. Indeed, recent US
legislation around so-called safe harbours and menu options has accelerated
this trend191919See the SECURE Act and associated policy issues, as well as
the original article by Bodie (1990) discussing the connection between
pensions as a form of retirement income insurance..
And yet most, if not all, of the published research articles and policy papers
that compute or reference these MWRs as a measure of cost efficiency have
focused on historical data, pricing information and quotes for life only
income annuities without any cash-refund features. Just in the year 2021 alone
we have come across a handful of articles that compute MWRs, all focusing
exclusively on life only income annuities. This isn’t just a matter of
empirical convenience or the availability of historical data. Most of the
highly cited theoretical papers in the annuity allocation literature, i.e.
deriving optimal dynamic strategies around annuitization, have also focused on
life only annuities202020Starting with Yaari (1965), as well as Davies (1981),
and more recently Kingston and Thorp (2005), Milevsky and Young (2007),
Horneff et al. (2009), Inkmann et al. (2011), Chai et al. (2011), Koijen et
al. (2011).. Any reference to a guaranteed period for the income annuity is
usually noted within the context of it being a fixed-income bond, and thus
irrelevant to the financial economics of longevity risk pooling.
And yet, we repeat once again that those are not the income annuities
consumers are buying. Rather, most of them want, demand and pay extra to
receive their money back over time. Our main point here is that the MWR
literature must progress to using cash-refund pricing algorithms, and our
paper offers encouragement and a path forward to those researchers. In
addition to our theoretical results, the appendix to this paper contains a
simple bisection-based algorithm (in R) that can be used to properly value
cash-refund income annuities under a Gompertz law of mortality. The mortality
curve can easily be discretized, modified or flattened.
### 2.3. Insurance Economics
The third and final strand of literature to which this paper contributes,
relates more generally to the so-called annuity puzzle and the very low levels
of voluntary annuitization observed among retirees, despite the classic Yaari
(1965) result which argues for full annuitization212121For the initial
statement and framing, as well as more recent empirical evidence on the extent
of the puzzle, and reasons a thin annuity market might be rational, see
Modigliani (1988), Davidoff et al. (2005), Brown et al. (2008), Benartzi et
al. (2011), Pashchenko (2013), Bommier and Le Grand (2014), and finally
Reichling and Smetters (2015).. The annuity puzzle literature, which is just
as vast as the above noted pension economic or actuarial science work, has
offered a variety of both classical (market frictions and incompleteness) and
behavioural (anchoring, mental accounting, loss aversion, etc.) reasons for
why consumer avoid annuitization. It continues to be an active area of
research.
Prima facie, the fact that from a very small group who actually choose to
annuitize, an even smaller group select the life only option further
exacerbates the puzzle. It’s a simple actuarial fact that per premium dollar,
the pure life only income annuity provide the highest level of income, the
greatest mortality credits and longevity pooling benefits to the annuitant.
And yet, recall that in Table 1, a mere 10% of annuitants selected the life
only income annuity. A full 90% of buyers opted for some form of guarantee,
and within that category a majority of those elected a cash-refund.
In fact, to add yet another layer of puzzlement, as interest rates have
declined over the last decade or so, the payout from income annuities have
declined across the entire spectrum of riders. The present value of everything
including the cost of retirement income goes up. Still, consumers have shifted
towards buying even more expensive income annuities that reduce their cash-
flow and income even further. At the risk of overusing the word, we pose this
as a further puzzle. Refunds are more expensive than ever, and continue to
increase in popularity.
But, the valuation model and results described in this paper allow us to shed
light on this matter by highlighting and isolating the embedded life insurance
component within the cash-refund annuity. As such, the demand for cash-refund
income annuities might be driven not-only by behavioural considerations noted
above, but perhaps by rational attempts by the annuitant (buyer) to signal to
the insurance company (seller) that they are not as healthy as a conventional
annuitant, purely interested in protecting against longevity risk. It also
might assist in maximizing a bequest value, more effectively compared to a
period certain222222See, for example, the article by Sheshinski (2008) as well
as the follow-up book referenced as Sheshinski (2010), in which he examines
what he calls refundable annuity options, albeit with a similar rationale.
option. Stated differently, the cash refund income annuity might be a better
deal for consumers, since it might reflect a lower level of anti-selection.
Furthermore, recent experimental evidence suggests that money back guarantees
are more likely to be selected as part of a so-called cushion effect.232323See
the paper by Knoller (2016), who makes an argument consistent with the IA
riders that consumers select, and see Boyer et al. (2020) for additional
survey-based evidence. And, while that would be an empirical question, we will
return to this in the conclusion of the paper when we discuss further
research. For now, we continue to Section 3 where we introduce and derive the
valuation and pricing technology.
## 3\. Actuarial analysis
### 3.1. Notation and terminology
In this paper we have tried as much as possible to adhere to standard
actuarial notation, but have deviated in a few places (elevating subscripts
and/or introducing superscripts) to reduce clutter and crowding.
* •
$a(x,r)$ is the price at age $x$, of a single premium life only (LO) income
annuity (IA) paying $\$1$ in continuous time, under a valuation rate $r$, and
with no loading. Because of the absence of loading, it agrees with the
actuarial present value of the LOIA. In other words, it is the present value
of the payment stream, discounted for both interest rates and mortality. Or,
by the law of large numbers, the cost per contract of hedging a large number
of identical such annuities. We will (quite often) abbreviate it by $a(x)$, or
occasionally just $a$, when the interest rate $r$ and/or age $x$ are clear
from the context.
* •
${a^{\star}}(x,r)$ is the price at age $x$, under a rate $r$, of an (unloaded)
IA paying $\$1$ in continuous time, and also guaranteeing a cash-refund (CR)
at death of: ${a^{\star}}(x,r)-t$, if death occurs at $t\leq{a^{\star}}(x,r)$.
Because there is no loading, it is also the actuarial present value (or value)
of this annuity. Naturally: ${a^{\star}}(x,r)>a(x,r)$. Once again, we might
use the simpler $a^{\star}(x)$ or just $a^{\star}$ when the context is clear.
* •
$a^{\circ}(x,r)$ is the price at age $x$, under a rate $r$, of an (unloaded)
IA paying $\$1$ in continuous time, guaranteeing at least $a^{\circ}(x,r)$
years of payments to the annuitant or beneficiary. Because there is no
loading, it is also the actuarial present value (or value) of this annuity.
This is the instalment refund income annuity (IRIA), which is effectively a
life-only IA, with an additional $a^{\circ}(x,r)$ years of guaranteed
payments. In Section 5 we prove that $a^{\ast}(x,r)>a^{\circ}(x,r)$ at any age
$x$ and any interest rate $r>0$, and in the current section focus on valuation
and numerical examples.
* •
An LOIA is $\tau$-period certain if the $1 paid continuously lasts for life or
time $\tau$, whichever is greater. We denote its price as $a(x,r,\tau)$. This
allows us to express an IRIA by adjusting $\tau$ recursively, that is,
$a^{\circ}=a^{\circ}(x,r)$ satisfies $a^{\circ}=a(x,r,a^{\circ})$.
Alternatively, $a(x,r,\tau)$ can be realized as the present value of $\$1$ for
time $\tau$ (i.e. $\frac{1-e^{-r\tau}}{r}$) plus a life annuity that only
starts payment at time $\tau$. The latter, called a deferred or delayed IA and
abbreviated as DIA, has been the focus of intense interest among pension
economists and has recently been granted special tax treatment in the
U.S.242424See Gong and Webb (2010), Pitacco (2016), Huang et al. (2017a),
Alexandrova and Gatzert (2019), Horneff et al. (2020), Habib et al. (2020),
and the regulatory exemptions granted to Qualified Longevity Annuity Contracts
(QLACs).
* •
$\pi\geq 0$ denotes the insurance loading or mark-up from value to price which
is applied to the IA by the insurance company, mainly to cover profits and
expenses, although in practice $\pi$ would include a safety margin to protect
against adverse mortality deviations. Practically speaking, loading a pure
actuarial premium by $\pi$, results in a present value of lifetime cash-flow
of: $C=\frac{P}{(1+\pi)a}$, where $P$ is the premium, or the annuitized
wealth. In other words, the price $\widehat{a}$ of a loaded LOIA will simply
$=(1+\pi)a$. The relationship between loaded and unloaded prices will be more
complicated in the case of a CRIA or an IRIA.
* •
$A(x,r)$ is the price at age $x$, of a single premium insurance policy paying
$\$1$ at death, under a valuation rate $r$. This implies that
$A(x,r)+ra(x,r)=1$, which can be shown via No Arbitrage arguments. Rearranging
this relationship leads to: $\frac{1}{a(x,r)}-r=\frac{A(x,r)}{a(x,r)}>0$,
which is deemed to be the mortality credits at age $x$.
* •
Finally, $T_{x}$ denotes the remaining lifetime random variable whose density
and distribution function are denoted $f_{x}(t)$ and $F_{x}(t)=\Pr[T_{x}\leq
t]=1-\,_{t}p_{x}$. The conditional survival probability under a Gompertz law,
which we will use for all our numerical examples, is:
$\,{}_{t}p_{x}=e^{-\int_{x}^{x+t}\lambda_{s}\,ds}=\exp\\{e^{(m-x)/b}(1-e^{t/b})\\}$,
where $m$ is the modal value and $b$ is the dispersion coefficient, and the
mortality hazard rate is: $\lambda_{s}=(1/b)e^{(s-m)/b}.$
### 3.2. Life Only IA
Following standard actuarial methodology,
(1) $a(x,r)\;=\;\int_{0}^{\infty}e^{-rt}\Pr[T_{x}\geq
t]dt\;=\;\int_{0}^{\infty}e^{-rt}\,(_{t}p_{x})dt,$
Under Gompertz mortality, this can be solved analytically252525For example,
see Milevsky (2006, Ch. 6). and leads to the expression:
(2)
$a(x,r)\;=\;\frac{b\Gamma(-rb,e^{(x-m)/b})}{\exp\\{(m-x)r-e^{(x-m)/b}\\}},$
where $\Gamma(A,B)$ is the incomplete Gamma function. For calibration and
comparison purposes we now display values for LOIA prices assuming two
different interest (valuation) rates: $r=2\%$ and $r=4\%$. We provide a short
R-script in the technical appendix, which comes from integrating equation (1),
and later will be compared against the instalment refund IA and cash refund IA
numerical values. Table 2 provides a range of numerical values of $a=a(x,r)$,
for $x=55,x=65,x=75$ under valuation rates $r=2\%$ and $r=4\%$. Likewise,
Figure 1 displays the LOIA price as a function of valuation rates $r$, at ages
$x=65,75$.
Table 2 Placed Here
Figure 1a (left panel) and Figure 1b (right panel) display the usual and
expected declining pattern for $a$, as valuation rates are increased. We will
return to this Figure when we discuss the cash-refund $a^{\star}$ and
instalment refund $a^{\circ}$ values.
Figure 1 Placed Here
### 3.3. Cash Refund IA
We now arrive at the “star” of this paper, and the focus of our attention, the
cash-refund income annuity (CRIA). The first thing to note is that its price
must be defined recursively, since the periodic cash-flow itself depends on
the original price. In the absence of loading, this can be expressed
mathematically as follows:
(3)
${a^{\star}}(x,r)\;=\;\int_{0}^{\infty}e^{-rs}\,(_{s}p_{x})ds\;+\;\int_{0}^{{a^{\star}}(x,r)}\left({a^{\star}}(x,r)\,-\,s\right)e^{-rs}\,(_{s}p_{x})\,\lambda_{(x+s)}\,ds,$
where the right hand side represents the actuarial present value of payments.
The annuitant receives $\$1$ for life, but if they die early, that is before
the entire ${a^{\star}}(x,r)$ has been returned, the beneficiary receives a
(type-of) life insurance payment consisting of the difference between the
price ${a^{\star}}(x,r)$ and payments received prior to death. The price is
constructed in dollars but also denotes an important time metric. Death before
age $y=x+{a^{\star}}(x,r)$ triggers a (declining) death benefit.
Notice how the first integral in equation (3) is the basic income annuity
price $a(x,r)$, and the second integral represents the non-negative life
insurance component. The CRIA price minus the LOIA price can also be expressed
as:
(4)
${a^{\star}}(x,r)-a(x,r)\;=\;\int_{0}^{{a^{\star}}(x,r)}\left({a^{\star}}(x,r)\,-\,s\right)e^{-rs}\,(_{s}p_{x})\,\lambda_{(x+s)}\,ds$
The technical appendix contains a short bisection-based algorithm for locating
numerical solutions to a modified version of this equation. For example, under
a Gompertz law of mortality with $m=90,b=10$ and $r=3\%$, the LOIA price at
age $x=65$ is ${a^{\star}}(65,0.03)=15.25$ while the CRIA price is
${a^{\star}}(65,0.03)=16.91$, both per dollar of lifetime income. Note, once
again, how in contrast to equation (2), the $a^{\star}$ appears on both sides
of equation (4) which leads to an implicit recusivity, and possibly to no
solution at all. Section 5 offers a proof of existence and uniqueness, and
shows that $a^{\star}(x,r)$ actually declines in both $x$ and $r$.
Nevertheless, at the age of $x=65$, under a $r=3\%$ valuation rate, the
difference between the CRIA and LOIA price is $1.66$ per dollar of lifetime
income, which is about $11\%$ more expensive. That is the cost of the money-
back guarantee, and the extra premium that buyers are paying for this rider.
Additional numerical values are presented in Table 3, where they can be
compared with the LOIA prices noted in Table 2. Once again, the cash-refund
feature is obviously a costly rider, and even more so in absolute as well as
relative terms, as valuation rates decline, (e.g. 2% versus 4%).
Table 3 Placed Here
In Section 5, we will show that (3) can be reformulated as the following;
(5)
$\int_{{a^{\star}}(x,r)}^{\infty}e^{-rt}{}(_{t}p_{x})\,dt=\int_{0}^{{a^{\star}}(x,r)}e^{-rt}r({a^{\star}}(x,r)-t){}(_{t}p_{x})\,dt.$
A particularly useful way to think about equation (5), as well as the
financial process by which the insurance company manages the CRIA payout, is
as follows. Initially, the annuitant’s entire premium ${a^{\star}}(x,r)$ is
placed into a so-called payout or phase-one account, which only contains the
sum that is refundable at death and is used to generate and payout the annuity
income up to time ${a^{\star}}(x,r)$. But, all interest income earned within
this (shrinking) phase-one account goes into – and is accumulated in – a
second phase-two account. Eventually, the insurer uses that phase-two account
to pay lifetime income after the phase-one account is emptied or depleted and
the formal guarantee period is over. Of course, insurance companies don’t
actually maintain two accounts for every annuitant, and in reality the entire
premium goes into one large general account, but it helps to think in this
bifurcated manner. With this framework, the LHS of equation (5) is the
discounted value of the phase-two payments. The RHS of equation (5) is the
income generated by the phase-one account, since $({a^{\star}}(x,r)-t)$ is the
account balance, and ${}_{t}p_{x}$ is the fraction of (initial) accounts
surviving.
### 3.4. Instalment Refund IA
A closer (and cheaper) cousin to the cash refund annuity (CRIA) is the
installment refund annuity (IRIA), under which payments continue to the
beneficiary until the entire premium is returned. Unlike the CRIA, there is no
lump-sum death benefit, rather payments continue as if the annuity included a
period certain of $a^{\circ}(x,r)$ years. Going back to the fundamental
valuation expression, payments for the first $t\leq a^{\circ}(x,r)$ years are
only discounted for interest (not mortality), and payments after
$t>a^{\circ}(x,r)$ are discounted for both interest and mortality.
Mathematically this can be expressed as:
$\displaystyle a^{\circ}(x,r)\;$
$\displaystyle=\;\int_{0}^{a^{\circ}(x,r)}e^{-rs}ds+\;\int_{a^{\circ}(x,r)}^{\infty}e^{-rs}\,(_{s}p_{x})\,ds$
(6)
$\displaystyle=\begin{cases}\frac{1}{r}(1-e^{-ra^{\circ}(x,r)})+\;\int_{a^{\circ}(x,r)}^{\infty}e^{-rs}\,(_{s}p_{x})\,ds,&r>0,\\\
a^{\circ}(x,r)+\;\int_{a^{\circ}(x,r)}^{\infty}\,(_{s}p_{x})\,ds,&r=0.\end{cases}$
IRIA values under the same parameters as before are shown in Table 4. Equation
(6) is recursive in $a^{\circ}(x,r)$, but unlike the CRIA price
${a^{\star}}(x,r)$ it is mathematically simpler. One can immediately see that
under $r=0$, $a^{\circ}(x,r)=\infty$, and the IA isn’t viable.
Table 4 Placed Here
Here is a scheme for locating $a^{\circ}(x,r)$, if you have a convenient or
closed-form expression, such as the Gompertz formula in equation (2), but for
an IA with a $\tau$-year period certain (PC). First, note that $a(x,r,\tau)$
is increasing in $\tau$, and is $>\tau$, when $\tau=0$. But,
$a(x,r,\tau)\rightarrow\frac{1}{r}<\tau$ as $\tau\rightarrow\infty$, as long
as $r>0$. So, increase $\tau$ until the curves cross and $a(x,r,\tau)=\tau$.
In Section 5 we will use a related argument to also show uniqueness, and to
handle loadings.
### 3.5. Insurance loading by $\pi$
Now, let’s return to the subject of insurance loadings. In contrast to the
LOIA computed in the prior subsection, it would be inappropriate and
technically incorrect to multiply ${a^{\star}}(x,r)$ by an insurance loading
of $(1+\pi)$ to convert them from unloaded to loaded prices. Why? Because by
charging or adding the extra loading, the insurer is obligated to pay a
greater death benefit, and for longer, even though their overall obligation to
pay $1 for life hasn’t changed. Using the above terminology, the phase-one
account must last longer. And, to preempt our main result, sometimes this
isn’t possible.
Let $\pi\geq 0$ denote the proportional insurance loading added to the
actuarial present value of a CRIA to yield its price. If $\widehat{a}^{\star}$
denotes the CRIA price, then the actuarial present value is
$\frac{\widehat{a}^{\star}}{1+\pi}$, so $\widehat{a}^{\star}$ must satisfy:
(7)
$\frac{\widehat{a}^{\star}}{1+\pi}=a\;+\;\int_{0}^{\widehat{a}^{\star}}\left(\widehat{a}^{\star}\,-\,s\right)e^{-rs}\,(_{s}p_{x})\,\lambda_{(x+s)}\,ds.$
For given values of $(x,m,b,\pi,r)$ it’s unclear a solution exists. And, as a
consequence of the results in Section 5, assuming Gompertz mortality with
parameters $m=90$ and $b=10$, you can not sell a cash-refund immediate annuity
(CRIA) if the valuation rate ($r$) falls below a certain level. Discounted
expectations won’t exist. The lowest viable valuation rate under which the
Cash Refund Income Annuity (CRIA) is still feasible will be denoted by
$r_{\pi,x}$, expressed in basis points – see Table 5. Thus, for example, with
Gompertz Mortality ($m=90$ and $b=10$) an inflation-adjusted CRIA would not be
feasible at $x=75$, under a loading of $\pi=15\%$, if real rates used for
pricing fall under $r_{0.15,75}=1.01\%$.
Table 5 Placed Here
Loaded prices for IRIAs are defined in the same way, so we have expressions
(or at least algorithms) for all three IAs. Figure 2 displays LOIA, IRIA and
CRIA prices assuming Gompertz mortality with $(m=90,b=10)$, and a valuation
interest rate of $r=2\%$. The dashed curves capture the prices without
loading, while the solid lines show the loaded prices. At younger ages
(visually under age 40) all three prices are nearly identical and the
guarantees or refunds don’t make much of a difference in value. But, at older
ages the IRIA and CRIA are (relative) more expensive.
Notice how at the age of $x=60$ the IRIA and CRIA prices are approximately 10%
higher than the LOIA prices. But, at age $x=75$ and beyond, when mortality
credits start to “kick in”, the premium for refunds can exceed 50% to 70% of
the LOIA price. And at higher ages, the CRIA price starts increasing with $x$,
and is eventually not viable.
Observe that ${a^{\star}}(x,r)>a^{\circ}(x,r)$ and
${\widehat{a}^{\star}}(x,r)>\widehat{a}^{\circ}(x,r)$ here. We will see in
Section 5 that these relations always hold. This is plausible in the unloaded
case, but seems much less obvious under loading. Note also that while the two
LOIA prices agree asymptotically as $x\to\infty$, in other words whether the
annuity is loaded for costs and profits, or sold with no loading at all, the
limiting price at advanced ages is the same. Yet, this is not the case for
IRIAs and we explore this fully in Section 5.
Figure 2 Placed Here
## 4\. Sensitivity, duration, and money’s worth ratio
### 4.1. Impact of age and rates
In this section we discuss the “sensitivities” of the life-only (LO),
instalment refund (IR) and cash-refund (CR) income annuity (IA) prices,
relative to changes in the underlying parameters age $x$ and valuation rate
$r$. Recall from section 3 that although we have a closed-form analytic
expression for the LO prices $a(x,r)$ in a Gompertz framework, the IR price
$a^{\circ}(x,r)$ and CR price ${a^{\star}}(x,r)$ can only be computed via
iteration. Those iterations also lead to a rather complex process for
computing partial derivatives, which among other interesting quantities are
the core ingredient of so-called duration and convexity calculations.
To complicate matters further, although for LO the loaded price
$\widehat{a}=(1+\pi)a$, is a simple multiple of the unloaded price $a$, that
is not the case for the other two annuities. For IRIA and CRIA the loaded
prices, which we denoted in Section 3 by $\widehat{a}^{\circ}$ and
$\widehat{a}^{\star}$, are no longer linear multiples $(1+\pi)$ of the
$a^{\circ}$ and ${a^{\star}}$. Therefore “sensitivities” will not scale
either, and if $r_{\pi,x}$ exceeds $r$ they might not exist at all, for the
CRIA.
Figure 3 displays the sensitivities of IA prices (loaded and unloaded) as a
function of age. Notice in panel (a) that (in the unloaded case), the
sensitivities $\frac{\partial}{\partial x}$ of all three IAs are consistently
negative, consistent with prices declining in age $x$. Interestingly, notice
how prior to the age of (approximately) 85, aging reduces the LO price at a
greater rate than the CR price (the derivative is negative and lower). But
after that age the situation is reversed and the reduction in the LOIA price
slows down while the CRIA reduces at faster rate. In contrast to the story
told in panel (a), the age derivative of the loaded CRIA price
$\widehat{a}^{\star}$ in panel (b) is consistently above that of the loaded
LOIA price $\widehat{a}$. In fact, and this once again ties into our earlier
result, at some age the derivative goes positive. Once again, this means that
older people will have to pay more (not less) for the same dollar of lifetime
income.
Figure 3 Placed Here
Turning to the impact of interest rates $r$, we wish to know which of the
three IAs has the largest or greatest percentage sensitivity to a change
$\Delta r$ in interest rates. Or, in the language of bond pricing, at any
given age, whose duration is highest? While we measured the sensitivity to age
$x$ as the straight partial derivative $\partial/\partial x$, as discussed
above, for valuation rates $r$ we will proceed in a manner that is analogous
with the bond literature. The new (key) symbol is $Dr[\cdot]$, which will
operate on either the loaded or unloaded prices. So, from here on, will focus
attention on the following mathematical expression:
(8) $Dr[a]:=\frac{-\partial a/\partial r}{a},$
and similarly for the other IAs. The above expression in (8) has been labeled
life annuity duration in the insurance literature262626See, for example,
Charupat et al. (2016). and we adhere to that definition in this paper. Its
convenience and use derives from the well-known approximation that:
(9) $\frac{a(x,r+\Delta r)}{a(x,r)}-1=-Dr[a(x,r)]\times\Delta r$
When applied to the standard life-only IA $a$, the life annuity duration
$Dr[a]$ “operator” will collapse to the well-known Maculay duration – a
weighted and discounted time average of cash flows. In fact, the loading
$(1+\pi)$ will cancel and the $Dr[a]$ is identical for the loaded and unloaded
prices. However, the $Dr[\cdot]$ will not correspond with the Macaulay
duration in the case of the IRIA and CRIA prices, once again due to the same
culprit, recursivity. We will get back to that in a moment, and despite the
popularity (and history) of Macaulay, we limit our analysis and comments to
duration defined as $Dr[\cdot]$.
The rationale for the $Dr[a]$ as our definition of life annuity duration –
even for the IRIA and CRIA – can be easily understood when it’s applied to a
simple (no mortality) present value (a.k.a. zero coupon bond) maturing at $1.
In that case $a:=e^{-rT}$ and $Dr[a]=T$, which is evidently the time until
receipt of the payment. This is why units of $Dr[a]$, and by extension
$D\lambda[a]$, are quoted in years. Moreover, if we replace $r$ by $r+\Delta
r$, the percentage change in the (new) value of $a$ can be approximated to
first order (Taylor expansion of $e^{\Delta T}$) by $\Delta rT$ percent. Now,
to continue developing intuition for our life annuity duration, let’s take the
case of a life only IA in which the force of mortality is constant and
lifetimes are exponentially distributed. The IA price is:
$a:=\int_{0}^{\infty}e^{-(r+\lambda)s}ds=(r+\lambda)^{-1}$, and from the
definition in (8), the $Dr[a]=a$. In this case, the percentage change in the
IA can be approximated by $-a\Delta r$, for small enough values of $\Delta r$.
For example, if $r=3\%$ and $\lambda=2\%$ (which is a life only IA issued at a
life expectancy of 50 years), the IA price is $20$ dollars and the life
annuity duration is (also) $20$ years. If we add $+25$ basis points to the
interest rate $r$ the new IA price is simply: $19.0476=1/(0.0525)$ which is a
decline of $4.76\%$. In contrast, using the life annuity duration
approximation, multiplying the $-Dr[a]=20$ by $\Delta r=0.0025$ is a change of
$-5\%$. We would like to obtain similar expressions (and approximations) for
the IRIA and CRIA prices.
As for the relationship between $Dr[a]$ and Macaulay duration we offer the
following observation. Regardless of the exact IA type, if cash flows from the
insurance company to the annuitant at a rate $\phi_{t}$ at time $t$, the
associated Macaulay duration is
$\frac{\int_{0}^{\infty}te^{-rt}\phi_{t}\,dt}{\int_{0}^{\infty}e^{-rt}\phi_{t}\,dt}$,
measured in units of time. In other words the arrival times $t$ are weighted
by the present values $e^{-rt}\phi_{t}$ of the associated cash flows. So, in
the case of an LOIA, if ${a}(r)=\int_{0}^{\infty}e^{-rt}\phi_{t}\,dt$,
Macaulay duration is $-\frac{1}{{a}}\frac{d{a}}{dr}$, which is consistent with
$Dr[{a}]$ noted above in (8). However, this is no longer the case for IRIA and
CRIA. Macaulay duration, as defined above, is simple to compute – just hold
${a^{\star}}$ constant in the right hand side of (3), while differentiating
with respect to $r$. This no longer equals $Dr[a]$, b/c changing $r$ does not
leave ${a^{\star}}$ constant, so the expression loses its interpretation as a
sensitivity to interest rates. Nevertheless, we will continue to report life
annuity duration in years.
### 4.2. Numerical examples
Here we provide some intuition for life annuity duration. For starters, it
should be clear from the left-hand panel of Figure 4, that at younger ages the
life annuity durations of all three unloaded products are virtually
indistinguishable from each other. The values of $Dr[a]$, as well as
$Dr[a^{\circ}]$ and $Dr[{a^{\star}}]$ decline with age $x$, but much more
rapidly for the LOIA. Indeed, the life annuity durations are in the vicinity
of 10-15 years around the retirement ages, which is what one might expect from
a fixed income product that pays “on average” until the period ends. Note that
at higher issue ages, and certainly during the traditional period of
retirement between age 65 and 85, the life annuity duration of the CR and IA
rider is 8-10 years higher (larger, more sensitive) than the LO version. This
depends on parameter values (in this case $m=90,b=10,r=2\%$) but is a feature
of all refunds at advanced ages. Higher duration has immediate and practical
applications for hedging and ALM.
Figure 4 Placed Here
Now, the situation is markedly different and rather counter-intuitive for the
loaded IA prices. The right-hand panel of Figure 4 display the
$Dr[\widehat{a}]$ for the LOIA, together with $Dr[\widehat{a}^{\circ}]$ for
the IRIA and $Dr[\widehat{a}^{\star}]$ for the CRIA. A number of odd things
can immediately be observed. First, at younger ages, the life annuity duration
for the IRIA and CRIA are now lower and under the LOIA curve. They actually
cross over – for these particular parameter values – somewhere around the age
of $x=40$. After that point the life annuity durations rank in the same
relative order as the unloaded prices. But, the oddities continue. While the
LOIA duration continues to decline, and is unaffected by the $\pi$ loading
which cancels out in the equation (derived in Section 5), the IRIA and CRIA
life annuity durations begin to increase somewhere around the age of $x=70$.
In fact, the CRIA duration – again, these are loaded prices – asymptotes
around the critical age at which the value no longer exists. Once again, we
refer readers to Section 5 to better understand these aspects of the
fundamental pricing equation.
### 4.3. Money’s Worth Ratio
We conclude this section with a brief discussion of the implications of our
work for the computation of the so-called Money’s Worth Ratio (MWR) which, as
we noted in the literature review, is an area of continued interest among
scholars in pension economics and finance. Generally speaking the MWR is
defined as the ratio of the theoretical model value and empirical market price
of an IA. If the measured ratio is less than one, which is usually the case
since market prices are loaded, then one is said (by an economist) to be
paying more than a fair price for the IA. If the ratio happens to be greater
than one (which is quite rare) the IA is considered to be a good deal (by an
economist) and if the ratio is exactly equal to one, the market price would be
perfectly fair. This way or approach to analyzing annuity market prices was
first introduced over 30 years ago by the economists Friedman and Warshawsky
(1990), has become the de facto standard in the literature and continues to be
used; see for example Poterba and Solomon (2021). Using our notation one can
think of the MWR as being approximately equal to $\frac{1}{1+\pi}$.
Now clearly the numerator in the MWR ratio – the theoretical model value of
the IA – requires an assumption for both mortality and discount rates, both of
which might be more complicated than the Gompertz $(m,b)$ law and a constant
valuation rate $r$. After all, one never really knows exactly what the
insurance company is assuming in their pricing. It is therefore more accurate
to present the MWR estimates as being conditional on a model. So, if $C$
denotes the annual cash-flow generated by the IA and $P$ denotes the market
premium or price, then $P/C$ is the empirical price per dollar of lifetime
income, which can be compared with the theoretical LOIA price $a$ or CRIA
price $a^{\star}$. With that in mind we will define the Gompertz MWR for both
the CRIA and the LOIA, naturally as follows:
${\tt MWR}^{\star}(x,r,m,b\mid
P,C)=\frac{a^{\star}(x,r,m,b)}{P/C},\;\;\;\;\;\;{\tt MWR}(x,r,m,b\mid
P,C)=\frac{a(x,r,m,b)}{P/C}$
Our objective is to estimate the MWR for LOIA and CRIA, to see if there is any
meaningful difference between them. Table 6 displays results and this is how
to interpret those numbers. A 65-year-old male who invested $P=\$100,\\!000$
into an IA in early July 2021, would have been entitled to (guaranteed)
approximately $C=\$5,\\!844$ per year for life, paid monthly, if he selected
the life only version (according to data from Fidelity Investments). But had
he selected the cash refund the payout would have been only $C=\$5,\\!280$ per
year, for reasons that should be evident by this point.
Table 6 Placed Here
Using those numbers, if we assume a Gompertz mortality model with $m=90$ years
and $b=10$ years, and a constant $r=2\%$ valuation rate, the MWR estimate of
the life only payout is exactly $0.996$ per premium dollar. To be clear, this
number is obtained by dividing the theoretical value $a(65,0.02,90,10)$ by the
empirical ratio $\frac{100000}{5844}=17.1157$. Note that we are not claiming
the insurance company uses this model and this valuation rate to price LOIAs.
Rather, under those assumptions the MWR would indicate perfect fairness to an
economist. In fact, we iterated $r$ until we converged to MWR=1.
But here is where it gets interesting. When we computed this ratio using the
same $(r,m,b)$ parameter assumptions at age $x=65$, but using the lower CRIA
payout of $C=\$5,\\!280$, the ${\tt MWR}^{\star}$ estimate is actually higher
than 1. In fact, the ${\tt MWR}^{\star}=1.03$ which might not seem like very
much, but is actually a rather large number within the money’s worth ratio
literature. More importantly, all the remaining rows in Table 6 tell the exact
same story. While the ${\tt MWR}(x,r,m,b)\approx 1$ for the LOIA, the ${\tt
MWR}^{\star}(x,r,m,b)>1$ for the CRIA. Why?
An answer to this question, or a possible explanation for this empirical fact
gets back to the economic essence of cash refund immediate annuities. Recall
that there were two components or integrals in the valuation of the CRIA price
$a^{\star}$. This first was the annuity payment of $1 for life, or the basic
$a$ price. On a stand-alone basis, the relevant mortality assumptions for that
component would be (healthy, anti-selected) annuitant mortality. But the
second component in the construction of $a^{\star}$ is associated with a life
insurance payment due upon (early) death. It’s unclear that portion should be
valued and discounted using annuitant mortality, given that it reflects a
payment upon death. Moreover, it could very well be that buyers who select a
cash refund (versus a life only) feature are in some way signalling they are
less healthy than average annuitants. This then induces the insurance company
to use less conservative (read: lower than $m=90$) values, which results in a
higher than unity ${\tt MWR}^{\star}$, computed relative to the conservative
(read: $m=90$) values.
In sum, although this is all rather speculative and demands further research,
it seems that when measured properly the MWR values are higher than life only
versus cash refund IAs. And, since they now form the majority of sales in the
U.S., we would urge the literature to focus more attention on CRIAs and their
possible role as a vehicle for satisfying bequest motives in a more cost-
efficient manner.
## 5\. Theorems and General Formulae
### 5.1. Theorems and proofs: CRIA with fair pricing
In this sub-section we provide a formal and proper statement of our theorems
and corresponding lemmas, as well as the proofs alluded to in the body of the
paper. We start by rewriting equation (3), in terms of the density $f(t)$ of
$T$:
(10)
${a^{\star}}=\int_{0}^{\infty}\frac{1-e^{-rt}}{r}f(t)\,dt+\int_{0}^{a^{\star}}({a^{\star}}-t)e^{-rt}f(t)\,dt.$
Here is an alternate version, written just in terms of ${}_{t}p_{x}$:
$\displaystyle{a^{\star}}$
$\displaystyle=\int_{0}^{\infty}e^{-rt}{}_{t}p_{x}\,dt+\int_{0}^{a^{\star}}\int_{s}^{a^{\star}}e^{-rt}(1+r({a^{\star}}-t))\,dtf(s)\,ds$
$\displaystyle=\int_{0}^{\infty}e^{-rt}{}_{t}p_{x}\,dt+\int_{0}^{a^{\star}}e^{-rt}(1+r({a^{\star}}-t))(1-{}_{t}p_{x})\,dt$
(11)
$\displaystyle={a^{\star}}+\int_{a^{\star}}^{\infty}e^{-rt}{}_{t}p_{x}\,dt-\int_{0}^{a^{\star}}e^{-rt}r({a^{\star}}-t){}_{t}p_{x}\,dt$
Or in other words,
(12)
$\int_{a^{\star}}^{\infty}e^{-rt}{}_{t}p_{x}\,dt=\int_{0}^{a^{\star}}e^{-rt}r({a^{\star}}-t){}_{t}p_{x}\,dt.$
This formula appeared earlier as (5) and, as noted at the time, can be
interpreted via making payments first from a phase-one and then from a phase-
two account.
Define functions $F(\alpha)=\int_{\alpha}^{\infty}e^{-rt}{}_{t}p_{x}\,dt$ and
$G(\alpha)=\int_{0}^{\alpha}e^{-rt}r(\alpha-t){}_{t}p_{x}\,dt$ of a real
variable $\alpha$. So by (12) we have that $a^{\star}$ is the value of
$\alpha$ making $F(a^{\star})=G({a^{\star}})$. Over the next few pages of this
section we plan to examine the dependence of these two expressions or terms on
some of the underlying parameters. In terms of notation, we will express these
functions as, for example, $F(\alpha;r)$ or $G(\alpha;x)$, but otherwise we
will not explicitly show the parameters.
###### Theorem 1.
Consider a CRIA with no loading, and let $r>0$.
1. (a)
There is a unique ${a^{\star}}(x)>0$ at which the CRIA is viable (ie
$F({a^{\star}})=G({a^{\star}})$);
2. (b)
$r{a^{\star}}(x)<1$
3. (c)
${a^{\star}}(x)$ is $\downarrow$ in $r$ with $\lim_{r\downarrow
0}{a^{\star}}(x)=\infty$, $\lim_{r\to\infty}r{a^{\star}}(x)=1$
4. (d)
Assume an increasing hazard rate. Then ${a^{\star}}(x)$ is $\downarrow$ in
$x$. If we also assume that $\lim_{x\to\infty}\lambda_{x}=\infty$ then
$\lim_{x\to\infty}{a^{\star}}(x)=0$.
###### Proof.
Note that $F(\alpha)$ is a decreasing function of $\alpha$, starting above 0
when $\alpha=0$ and $\to 0$ when $\alpha\to\infty$. The derivative
$\frac{\partial
G(\alpha)}{\partial\alpha}=r\int_{0}^{\alpha}e^{-rt}{}_{t}p_{x}\,dt$ is
increasing in $\alpha$, so $G$ is a convex increasing function, starting at 0.
Therefore $F$ and $G$ will cross at a unique value of $\alpha$, which
therefore defines $a^{\star}$. This shows item (a) in the above theorem.
To prove (b), we use that ${}_{t}p_{x}$ is $\downarrow$ in $t$. Therefore at
$\alpha={a^{\star}}(x)$ we have that:
${}_{a^{\star}}p_{x}\frac{e^{-r{a^{\star}}}}{r}={}_{a^{\star}}p_{x}\int_{a^{\star}}^{\infty}e^{-rt}\,dt>F({a^{\star}})\\\
=G({a^{\star}})>{}_{a^{\star}}p_{x}\int_{0}^{a^{\star}}e^{-rt}r({a^{\star}}-t)\,dt={}_{a^{\star}}p_{x}[{a^{\star}}-\frac{1-e^{-r{a^{\star}}}}{r}].$
This immediately implies that ${a^{\star}}r<1$. Turning to item (c), if
$r\downarrow 0$, then $\frac{\partial G}{\partial\alpha}\to 0$ for each
$\alpha$, so $G\to 0$ pointwise. That is not true of $F$, so in fact
${a^{\star}}(x)\to\infty$. For $r\to\infty$ we have $ra_{x}<r{a^{\star}}(x)<1$
by (b). Because ${}_{t}p_{x}\to 0$ as $t\to\infty$ it is simple to show that
$ra_{x}\to 1$. Therefore $r{a^{\star}}\to 1$. To show that ${a^{\star}}(x)$
decreases with $r$, observe that $\alpha={a^{\star}}(x)$ is the point where
the decreasing function $F(\alpha;r)-G(\alpha;r)$ reaches 0. By (b) of Lemma 2
below, increasing $r$ will lower the path $F(\alpha;r)-G(\alpha;r)$ for
$\alpha\in[0,\frac{1}{r}]$. This interval contains ${a^{\star}}(x)$ by (b), so
this forces the zero of $F-G$ to the left. This shows (c). Now assume an
increasing hazard rate. The first statement in part (d) follows as above, this
time using (c) of Lemma 2. Finally, suppose that the hazard rate increases
without bound. Fix $\epsilon>0$. Let $\lambda=\lambda_{x+\epsilon}$. Then
${}_{t}p_{x}>e^{-\lambda t}$ for $t<\epsilon$ and
${}_{t}p_{x}<{}_{\epsilon}p_{x}e^{-\lambda(t-\epsilon)}$ for $t>\epsilon$.
Therefore
$F(\epsilon)<\int_{\epsilon}^{\infty}{}_{\epsilon}p_{x}e^{-rt}e^{-\lambda(t-\epsilon)}\,dt=\frac{e^{-r\epsilon}}{r+\lambda}{}_{\epsilon}p_{x}=o(\frac{1}{r+\lambda})$
since ${}_{\epsilon}p_{x}\to 0$ when $x\to\infty$. Likewise
$G(\epsilon)>\int_{0}^{\epsilon}e^{-rt}r(\epsilon-t)e^{-\lambda
t}\,dt=\frac{r}{r+\lambda}\Big{[}\epsilon-\frac{1}{r+\lambda}(1-e^{-(r+\lambda)\epsilon})\Big{]}\sim\frac{r\epsilon}{r+\lambda}$
when $x\to\infty$, since also $\lambda\to\infty$. In particular,
$F(\epsilon)<G(\epsilon)$ for $x$ sufficiently large, which implies that
${a^{\star}}(x)<\epsilon$. Since $\epsilon$ was arbitrary, in fact
${a^{\star}}(x)\to 0$. ∎
The above argument relied on the following result, which we now prove.
###### Lemma 2.
Consider $F$ and $G$ as above. Then
1. (a)
$\frac{\partial F(\alpha)}{\partial\alpha}<0<\frac{\partial
G(\alpha)}{\partial\alpha}$.
2. (b)
$\frac{\partial F(\alpha;r)}{\partial r}<0<\frac{\partial
G(\alpha;r)}{\partial r}$ for $0<\alpha<\frac{1}{r}$.
3. (c)
Assume an increasing hazard rate. Then $\frac{\partial F(\alpha;x)}{\partial
x}<\frac{\partial G(\alpha;x)}{\partial x}$ at $\alpha={a^{\star}}(x)$.
###### Proof.
We’ve already seen the argument for part (a), at the beginning of the proof of
Theorem 1. For part (b), $\frac{\partial F(\alpha;r)}{\partial
r}=-\int_{\alpha}^{\infty}te^{-rt}{}_{t}p_{x}\,dt<0$ and $\frac{\partial
G(\alpha;r)}{\partial
r}=\int_{0}^{\alpha}e^{-rt}(\alpha-t){}_{t}p_{x}[1-rt]\,dt$. Since
$r\alpha<1$, the integrand is $>0$, so $\frac{\partial G(\alpha;r)}{\partial
r}>0$.
For part (c) we have
$\displaystyle\frac{\partial(F-G)}{\partial x}$
$\displaystyle=\int_{\alpha}^{\infty}e^{-rt}{}_{t}p_{x}[\lambda_{x}-\lambda_{x+t}]\,dt-\int_{0}^{a^{\star}}e^{-rt}r(\alpha-t){}_{t}p_{x}[\lambda_{x}-\lambda_{x+t}]\,dt$
$\displaystyle=\lambda_{x}F-\lambda_{x}G+\int_{0}^{\alpha}e^{-rt}r(\alpha-t){}_{t}p_{x}\lambda_{x+t}\,dt-\int_{\alpha}^{\infty}e^{-rt}{}_{t}p_{x}\lambda_{x+t}\,dt$
$\displaystyle=\int_{0}^{a^{\star}}e^{-rt}r({a^{\star}}-t){}_{t}p_{x}\lambda_{x+t}\,dt-\int_{a^{\star}}^{\infty}e^{-rt}{}_{t}p_{x}\lambda_{x+t}\,dt$
at $\alpha=a^{\star}$ since $F(a^{\star})=G(a^{\star})$. Assume an increasing
hazard rate. Then
$\int_{0}^{a^{\star}}e^{-rt}r({a^{\star}}-t){}_{t}p_{x}\lambda_{x+t}\,dt<\int_{0}^{a^{\star}}e^{-rt}r({a^{\star}}-t){}_{t}p_{x}\lambda_{x+{a^{\star}}}\,dt=\lambda_{x+{a^{\star}}}G(a^{\star})$
while
$\int_{a^{\star}}^{\infty}e^{-rt}{}_{t}p_{x}\lambda_{x+t}\,dt>\int_{a^{\star}}^{\infty}e^{-rt}{}_{t}p_{x}\lambda_{x+{a^{\star}}}\,dt=\lambda_{x+{a^{\star}}}F(a^{\star}).$
Therefore the difference is
$<\lambda_{x+{a^{\star}}}(G(a^{\star})-F(a^{\star}))=0$. ∎
Note that the statement of item (b) above will be strengthened below (see
Lemma 5), but we prefer to keep the short proof given above, for the case
considered here. Note that with $r=0$ the CRIA is clearly non-viable: We have
$G=0$ while $F>0$. For $r<0$ things are even worse – $F>0$ while $G<0$. Note
also that (c) of the Theorem need not hold, without the assumption of an
increasing hazard rate. For example, if the hazard rate decreases, the proof
of the Lemma implies that $\frac{\partial[F(\alpha;x)-G(\alpha;x)]}{\partial
x}>0$ at $\alpha=a^{\star}$, and therefore $\frac{d{a^{\star}}}{dx}>0$. Though
${a^{\star}}$ declines with $x$, it cannot decline too fast, as the next Lemma
shows. Let $y=x+{a^{\star}}(x)$ be the age at which the cash refund expires.
Then:
###### Lemma 3.
For a fairly priced CRIA, $y$ increases with $x$.
###### Proof.
Re-express $F$ and $G$ as
$\tilde{F}(y;x)=\int_{y-x}^{\infty}e^{-rt}{}_{t}p_{x}\,dt,\qquad\tilde{G}(y;x)=r\int_{0}^{y-x}e^{-rt}(y-x-t){}_{t}p_{x}\,dt.$
As before, we let $y=y(x)$ make $\tilde{F}(y;x)-\tilde{G}(y;x)=0$. By implicit
differentiation,
$\frac{dy}{dx}=-\frac{\frac{\partial[\tilde{F}-\tilde{G}]}{\partial
x}}{\frac{\partial[\tilde{F}-\tilde{G}]}{\partial y}}$
at $y=y(x)$. But $\frac{\partial\tilde{F}}{\partial y}<0$ and
$\frac{\partial\tilde{G}}{\partial y}>0$. And using integration by parts,
$\frac{\partial\tilde{F}}{\partial
x}=e^{-r(y-x)}{}_{y-x}p_{x}+\int_{y-x}^{\infty}e^{-rt}{}_{t}p_{x}[\lambda_{x}-\lambda_{x+t}]\,dt=(r+\lambda_{x})\tilde{F}$
and
$\displaystyle\frac{\partial\tilde{G}}{\partial x}$
$\displaystyle=-r\int_{0}^{y-x}e^{-rt}{}_{t}p_{x}\,dt+r\int_{0}^{y-x}e^{-rt}(y-x-t){}_{t}p_{x}[\lambda_{x}-\lambda_{x+t}]\,dt$
$\displaystyle=-r\int_{0}^{y-x}e^{-rt}{}_{t}p_{x}\,dt+\lambda_{x}\tilde{G}-r(y-x)-r\int_{0}^{y-x}e^{-rt}[-r(y-x-t)-1]{}_{t}p_{x}\,dt$
$\displaystyle=(r+\lambda_{x})\tilde{G}-r(y-x).$
So the desired inequality holds. ∎
### 5.2. Theorems and proofs: CRIA with loading
Next consider viability in the presence of loading. In actuarial terminology,
loading is the percentage increase applied to the basic hedging cost to obtain
the price charged consumers, with the difference going to fund expenses,
capital reserves, or insurer profits. Recall that we assigned the symbol $\pi$
for the proportional loading, and used that in the body of the paper and for
the numerical examples in Table 5, with $\pi=5\%,15\%,25\%$.
###### Theorem 4.
Consider a CRIA with loading $\pi$.
1. (a)
If $\frac{\pi}{1+\pi}<ra_{x}$ then there is a unique
${\widehat{a}^{\star}}(x)$ at which the CRIA is viable. Moreover,
${\widehat{a}^{\star}}(x)$ increases with $\pi$ and decreases with $r$.
2. (b)
If $\frac{\pi}{1+\pi}\geq ra_{x}$ there is no ${\widehat{a}^{\star}}$ making
the CRIA viable.
3. (c)
For each $\pi>0$ and each $x$, there is an $r_{\pi,x}$ such that the CRIA is
viable when $r>r_{\pi,x}$ but not when $r\leq r_{\pi,x}$. For fixed $x$,
$r_{\pi,x}\to 0$ when $\pi\downarrow 0$, and $r_{\pi,x}\to\infty$ when
$\pi\uparrow\infty$. If $r\downarrow r_{\pi,x}$ then
${\widehat{a}^{\star}}(x)\to\infty$. If $r\to\infty$ then
$r{\widehat{a}^{\star}}(x)\to 1+\pi$.
4. (d)
Assume a hazard rate that increases without bound. For each $\pi>0$ and $r>0$
there is an $x_{\pi,r}$ such that The CRIA is viable when $x<x_{\pi,r}$ but
not when $x\geq x_{\pi,r}$. If $x\uparrow x_{\pi,r}$ then
${\widehat{a}^{\star}}(x)\to\infty$.
###### Proof.
Loading changes the LHS of (11) into $\frac{1}{1+\pi}\widehat{a}^{\star}$, and
it therefore adds $\frac{\pi}{1+\pi}\widehat{a}^{\star}$ to the LHS of (12).
Let $\delta=\frac{\pi}{1+\pi}$. So the CRIA is viable at price
${\widehat{a}^{\star}}$ if
$\delta{\widehat{a}^{\star}}+F({\widehat{a}^{\star}})=G({\widehat{a}^{\star}})$.
Then $G(\alpha)\leq r\alpha\int_{0}^{\alpha}e^{-rt}{}_{t}p_{x}\,dt<r\alpha
a_{x}$ and $\delta\alpha+F(\alpha)>\alpha\delta$. In particular, if
$\delta\geq ra_{x}$ then the curves cannot cross, so no
${\widehat{a}^{\star}}$ can be viable. This shows (b). We know that
$F(\alpha)-G(\alpha)$ starts positive at $\alpha=0$, and decreases forever.
Its asymptotic slope is $-ra_{x}$. In particular, if $\delta<ra_{x}$ then it
must cross the curve $-\delta\alpha$, so there is a viable
${\widehat{a}^{\star}}$. The argument also implies that if we let
$\delta\uparrow ra_{x}$, any ${\widehat{a}^{\star}}$ that achieves viability
will $\to\infty$. Because $-\delta\alpha$ decreases with $\delta$, we also
conclude that the viable ${\widehat{a}^{\star}}$ increases with $\delta$, and
hence with $\pi$.
To see uniqueness of ${\widehat{a}^{\star}}$, observe
$F^{\prime\prime}(\alpha)-G^{\prime\prime}(\alpha)>0$ so $F-G$ is convex. So
if $F(\alpha)-G(\alpha)$ crosses the line $-\delta\alpha$ twice, then after
the second crossing it must stay above that line. This is not compatible with
the asymptotic slopes, so in fact there is a unique crossing. To show that
${a^{\star}}(x)$ decreases with $r$, we will use Lemma 5 below.
It shows that raising $r$ will lower the curve $F(\alpha;r)-G(\alpha;r)$. This
pushes down the value of ${\widehat{a}^{\star}}$ at which this curve crosses
the line $-\delta\alpha$, which completes the proof of (a). To address (c) we
need to understand how the function $\rho(r)=ra_{x}$ varies with $r$. We have
$\rho(0)=0$, and integration by parts shows that
$\rho(r)=1-\int_{0}^{\infty}e^{-rt}{}_{t}p_{x}\,\lambda_{x+t}\,dt$. This
increases with $r$, converging to 1 as $r\to\infty$. The existence and
properties of $r_{\pi,x}$ now follow from our earlier conclusions. To find the
asymptotics as $r\to\infty$, observe that the viable ${\widehat{a}^{\star}}$
satisfies $r[F({\widehat{a}^{\star}})-G({\widehat{a}^{\star}})]=-\delta
r{\widehat{a}^{\star}}$. Letting $B=r{\widehat{a}^{\star}}$, and changing
variables in the integrals, this becomes that
$\int_{B}^{\infty}e^{-s}{}_{\frac{s}{r}}p_{x}\,ds-\int_{0}^{B}e^{-s}(B-s){}_{\frac{s}{r}}p_{x}\,ds=-\delta
B.$
Sending $r\to\infty$ makes this asymptotically
$\int_{B}^{\infty}e^{-s}\,ds-\int_{0}^{B}e^{-s}(B-s)\,ds=-\delta B$, which
simplifies to $1-B=-\delta B$. So $B\to\frac{1}{1-\delta}=1+\pi$ which
completes the proof of (c). Under the assumptions of (d), if we increase $x$
then $a_{x}$ decreases, and $a_{x}\to 0$ when $x\to\infty$. The conclusions of
(d) now follow from our earlier statements. ∎
The above argument used the following result, which we now prove. It is a
strengthening of (b) of Lemma 2.
###### Lemma 5.
Consider $F(\alpha;r)$ and $G(\alpha;r)$ as above. Then $\frac{\partial
F}{\partial r}<0<\frac{\partial G}{\partial r}$ for every $\alpha>0$.
###### Proof.
As in Lemma 2, $\frac{\partial F(\alpha;r)}{\partial
r}=-\int_{\alpha}^{\infty}te^{-rt}{}_{t}p_{x}\,dt<0$. Let
$b={}_{\frac{1}{r}}p_{x}$. Since ${}_{t}p_{x}$ decreases with $t$,
$\frac{\partial G(\alpha;r)}{\partial
r}=\int_{0}^{\alpha}e^{-rt}(\alpha-t){}_{t}p_{x}[1-rt]\,dt>\int_{0}^{\alpha}be^{-rt}(\alpha-t)[1-rt]\,dt.$
The antiderivative required is
$-be^{-rt}\Big{(}\frac{1}{r^{2}}+[\frac{1}{r}-{\alpha}]t+t^{2}\Big{)}$, making
the above expression $=\frac{b}{r^{2}}\Big{(}1-e^{-r\alpha}(1+\alpha
r)\Big{)}$. This is $\geq 0$ since $e^{y}\geq 1+y$. ∎
### 5.3. Theorems and proofs: IRIA & Loading
Define
$H(\alpha)=\frac{1}{r}(1-e^{-r\alpha})+\int_{\alpha}^{\infty}e^{-rs}\,(_{s}p_{x})\,ds.$
Recalling (6), an unloaded IRIA is viable at price $a^{\circ}$ if
${a^{\circ}}=H({a^{\circ}})$. More generally, in the presence of loading by
$\pi$ , the relevant equation is that
(13) $\frac{1}{1+\pi}\widehat{a}^{\circ}=H({\widehat{a}^{\circ}}).$
###### Theorem 6.
Consider an IRIA with loading $\pi\geq 0$. Let $r>0$.
1. (a)
There is a unique ${\widehat{a}^{\circ}}(x)$ at which the IRIA is viable.
${\widehat{a}^{\circ}}(x)$ increases with $\pi$.
2. (b)
$r{\widehat{a}^{\circ}}(x)<1+\pi$.
3. (c)
${\widehat{a}^{\circ}}(x)$ is $\downarrow$ in $r$. It $\to\infty$ as
$r\downarrow 0$, and $r{\widehat{a}^{\circ}}(x)\to 1+\pi$ as $r\to\infty$.
4. (d)
Assume an increasing hazard rate. Then ${\widehat{a}^{\circ}}(x)$ is
$\downarrow$ in $x$. $\lim_{x\to\infty}{\widehat{a}^{\circ}}(x)>0$ if $\pi>0$.
Whereas if the hazard rate is also unbounded and $\pi=0$, then
$\lim_{x\to\infty}{a^{\circ}}(x)=0$.
###### Proof.
$H(0)>0$ and $H^{\prime}(\alpha)=e^{-r\alpha}(1-{}_{\alpha}p_{x})<1$. So the
viable ${a^{\circ}}$ exists and is unique when $\pi=0$. To see this in the
case $\pi>0$, rewrite (13) as
$\frac{1}{1+\pi}a^{\circ}-\frac{1-e^{-r{a^{\circ}}}}{r}=\int_{a^{\circ}}^{\infty}e^{-rs}{}_{s}p_{x}\,ds.$
Define $u(\alpha)=\frac{1}{1+\pi}\alpha-\frac{1-e^{-r{\alpha}}}{r}$ and
$v(\alpha)=\int_{\alpha}^{\infty}e^{-rs}{}_{s}p_{x}\,ds$, so that the above
equation becomes that $u(a^{\circ})=v(a^{\circ})$. Then $v(\alpha)$ is
$\downarrow$ in $\alpha$ and $>0$. The function $u$ starts at 0, initially
decreases, and then increases forever (and $\to+\infty$). Therefore they
cross. Moreover, by the time the two paths cross, we’re looking at the
intersection of an increasing path with a decreasing one, and there can only
be one such crossing. This shows (a).
Now observe that
$H(\alpha)<\frac{1-e^{-r\alpha}}{r}+{}_{\alpha}p_{x}\int_{\alpha}^{\infty}e^{-rt}\,dt=\frac{1-e^{-\alpha
r}[1-{}_{\alpha}p_{x}]}{r}<\frac{1}{r}$. Therefore
$\alpha_{0}=\frac{1+\pi}{r}$ satisfies
$H(\alpha_{0})<\frac{1}{r}=\frac{1}{1+\pi}\alpha_{0}$. So
${\widehat{a}^{\circ}}(x)<\alpha_{0}$, showing (b). Turning to (c),
$\frac{\partial H(\alpha;r)}{\partial
r}=-\int_{0}^{\alpha}te^{-rt}\,dt-\int_{\alpha}^{\infty}te^{-rt}{}_{t}p_{x}\,dt<0$.
So raising $r$ lowers the path $H$ and so decreases its crossing
${\widehat{a}^{\circ}}(x)$ with the path $\frac{1}{1+\pi}\alpha$. When $r\to
0$, we have $u(\alpha)\to-\frac{\pi}{1+\pi}\alpha\leq 0$, while $v(\alpha)$
stays bounded from 0 (for any fixed $\alpha$). This forces
${\widehat{a}^{\circ}}(x)\to\infty$.
When $r\to\infty$ we have $u(\alpha)\to\frac{1}{1+\pi}\alpha$ while
$v(\alpha)\to 0$, which forces ${\widehat{a}^{\circ}}(x)\to 0$. More
precisely, rewriting $ru({\widehat{a}^{\circ}})=rv({\widehat{a}^{\circ}})$ in
terms of $\beta=r{\widehat{a}^{\circ}}$ yields
$\frac{1}{1+\pi}\beta-1+e^{-\beta}=\int_{\beta}^{\infty}e^{-s}{}_{\frac{s}{r}}p_{x}\,ds\to
e^{-\beta}$
as $r\to\infty$. This simplifies to $\frac{1}{1+\pi}\beta\to 1$, which shows
(c).
Assume a rising hazard rate. Observe that function $u(\alpha)$ does not vary
with $x$, while the function $v(\alpha)$ declines with $x$ As above, this
forces the crossing ${\widehat{a}^{\circ}}(x)$ to decrease as $x$ rises. When
$\pi>0$, we have $u(\alpha)\leq 0$ for $\alpha$ in some interval $[0,z]$ that
does not depend on $x$. Therefore ${\widehat{a}^{\circ}}(x)>z$ for every $x$.
But if $\delta=0$ then $u(\alpha)>0$ for $\alpha>0$. If the hazard rate rises
without bound then $v(\alpha)\downarrow 0$ when $x\to\infty$, which forces
${a^{\circ}}(x)$ to $\to 0$. ∎
Note that the behaviour of a loaded IRIA at advanced ages (ie approaching a
non-zero constant) is quite different from that of a CRIA (blowing up) or a
LOIA (approaching 0).
### 5.4. Sensitivities
For completeness, we record general formulas for the various derivatives
needed to compute sensitivities, which were then evaluated numerically for
tables and figures.
* •
CRIA
$\frac{d{\widehat{a}^{\star}}}{dr}=-\frac{\partial(\frac{\pi}{1+\pi}\alpha+F-G)}{\partial
r}/\frac{\partial(\frac{\pi}{1+\pi}\alpha+F-G)}{\partial\alpha}\Big{|}_{\alpha=\widehat{a}^{\star}}$,
where
$\frac{\partial(F-G)}{\partial\alpha}=-e^{-r\alpha}{}_{\alpha}p_{x}-r\int_{0}^{\alpha}e^{-rt}{}_{t}p_{x}\,dt$
and $\frac{\partial(F-G)}{\partial
r}=-\int_{\alpha}^{\infty}te^{-rt}{}_{t}p_{x}\,dt-\int_{0}^{\alpha}e^{-rt}(\alpha-t){}_{t}p_{x}[1-rt]\,dt$.
And at $\alpha=\widehat{a}^{\star}$, $F-G=-\frac{\pi}{1+\pi}\alpha$.
* •
CRIA
$\frac{d{\widehat{a}^{\star}}}{dx}=-\frac{\partial(\frac{\pi}{1+\pi}\alpha+F-G))}{\partial
x}/\frac{\partial(\frac{\pi}{1+\pi}\alpha+F-G)}{\partial\alpha}\Big{|}_{\alpha=\widehat{a}^{\star}}$,
where $\frac{\partial(F-G)}{\partial
x}=\int_{0}^{\alpha}e^{-rt}r(\alpha-t){}_{t}p_{x}\lambda_{x+t}\,dt-\int_{\alpha}^{\infty}e^{-rt}{}_{t}p_{x}\lambda_{x+t}\,dt+\lambda_{x}[F-G]$,
which after applying integration by parts
$=r\alpha-e^{-r\alpha}{}_{\alpha}p_{x}-r\int_{0}^{\alpha}e^{-rt}{}_{t}p_{x}\,dt+(r+\lambda_{x})(F-G)$.
* •
LOIA $\frac{da_{x}}{dr}=-\int_{0}^{\infty}te^{-rt}{}_{t}p_{x}\,dt$. And the
loaded LOIA price is $\widehat{a}_{x}=(1+\pi)a_{x}$
* •
LOIA $\frac{da_{x}}{dx}=-(1-[r+\lambda_{x}]a_{x})$
* •
IRIA $\frac{d{\widehat{a}^{\circ}}}{dr}=\frac{\partial H}{\partial
r}/\Big{[}\frac{1}{1+\pi}-\frac{\partial
H}{\partial\alpha}\Big{]}\Big{|}_{\alpha=\widehat{a}^{\circ}}$, where
$\frac{\partial H}{\partial r}=\frac{e^{-r\alpha}(1+\alpha
r)-1}{r^{2}}-\int_{\alpha}^{\infty}te^{-rt}{}_{t}p_{x}\,dt$, and
$\frac{\partial H}{\partial\alpha}=e^{-r\alpha}(1-{}_{\alpha}p_{x})$. And at
$\alpha=\widehat{a}^{\circ}$, $H=\frac{1}{1+\pi}\alpha$.
* •
IRIA $\frac{d{\widehat{a}^{\circ}}}{dx}=\frac{\partial H}{\partial
x}/\Big{[}\frac{1}{1+\pi}-\frac{\partial
H}{\partial\alpha}\Big{]}\Big{|}_{\alpha=\widehat{a}^{\circ}}$, where
$\frac{\partial H}{\partial
x}=\int_{\alpha}^{\infty}e^{-rs}{}_{s}p_{x}[\lambda_{x}-\lambda_{x+s}]\,ds=-e^{-r\alpha}{}_{\alpha}p_{x}+(r+\lambda_{x})\int_{\alpha}^{\infty}e^{-rs}{}_{s}p_{x}\,ds$.
### 5.5. Theorems and proofs: CRIA dominates IRIA
###### Theorem 7.
We have ${\widehat{a}^{\star}}>{\widehat{a}^{\circ}}>\widehat{a}_{x}$ for any
$\pi\geq 0$ and $r>0$.
###### Proof.
Recall that $F(\alpha)=\int_{\alpha}^{\infty}e^{-rt}{}_{t}p_{x}\,dt$
represents the insurer’s liability due to annuitants who live beyond age
$\alpha$. We imagine that the purchase price $\alpha$ is deposited into what
we earlier called the phase-one account, which is used for the first $\alpha$
of payments, while interest earned goes into the phase-two account. Loading
requires the latter to cover payments to the insurer, whose present value
amounts to $\frac{\pi}{1+\pi}\alpha$, in addition to the liability
$F(\alpha)$. If $G_{i}(\alpha)$ represents the present value of interest
generated by the payout account (with $i=1,2,3$ corresponding respectively to
the LOIA, IRIA, and CRIA versions), then each respective price is determined
by matching this interest against liabilities, ie by solving
$\frac{\pi}{1+\pi}\alpha+F(\alpha)=G_{i}(\alpha)$. Since each $G_{i}(0)=0$ and
$F(0)>0$, the desired inequality will follow if we can show that
$G_{1}(\alpha)>G_{2}(\alpha)>G_{3}(\alpha)$ for every $\alpha$. For a CRIA,
$G_{3}(\alpha)=\int_{0}^{\alpha}e^{-rt}r(\alpha-t){}_{t}p_{x}\,dt$ (which was
earlier denoted simply $G(\alpha)$). For an IRIA,
$G_{2}(\alpha)=\alpha-\int_{0}^{\alpha}e^{-rs}\,ds=\alpha-\frac{1-e^{-r\alpha}}{r}$.
For a LOIA, $G_{1}(\alpha)=\alpha-\int_{0}^{\alpha}e^{-rt}{}_{t}p_{x}\,dt$.
Because ${}_{t}p_{x}<1$, it is clear from the integrals that
$G_{1}(\alpha)<G_{2}(\alpha)$. For the same reason,
$G_{3}(\alpha)<\int_{0}^{\alpha}e^{-rt}r(\alpha-t)\,dt$, which is easily seen
to $=G_{2}(\alpha)$. ∎
### 5.6. Macaulay Duration
As described earlier, our definition of duration differs from traditional
Macaulay duration in the case of CRIAs and IRIAs. But for completeness, we
give the Macaulay versions here. If cash flows at rate $\phi_{t}$ at time $t$,
the associated Macaulay duration is
$\frac{\int_{0}^{\infty}te^{-rt}\phi_{t}\,dt}{\int_{0}^{\infty}e^{-rt}\phi_{t}\,dt}$,
measured in units of time. In other words the arrival times $t$ are weighted
by the present values $e^{-rt}\phi_{t}$ of the cash flows. So if
$M(r)=\int_{0}^{\infty}e^{-rt}\phi_{t}\,dt$, life annuity duration is
$-\frac{1}{M}\frac{\partial M}{\partial r}$. Note that the cash flows
$\phi_{t}$ are being held constant here, rather than adjusting with $r$. In
the case of a CRIA, the present value of cash flows is
$M(r,{\widehat{a}^{\star}})={\widehat{a}^{\star}}+F({\widehat{a}^{\star}},r)-G({\widehat{a}^{\star}},r)$
where ${\widehat{a}^{\star}}$ is obtained by solving
$\frac{1}{1+\pi}\widehat{a}^{\star}=M(r,{\widehat{a}^{\star}})$. Therefore the
formula for Macaulay duration, as defined above, now becomes
$-\frac{1+\pi}{{\widehat{a}^{\star}}}\frac{\partial M}{\partial r}$. This is
now easily computed. We can understand the asymptotics as $x$ increases to the
maximum feasible age. As that happens, we know
${\widehat{a}^{\star}}\to\infty$. Based on the discussion in Section 3 of the
paper, life annuity duration converges to
$(1+\pi)\int_{0}^{\infty}e^{-rt}{}_{t}p_{x}(1-rt)\,dt>0$. To see the
inequality, observe that the sign of the integrand and the $\downarrow$ nature
of ${}_{t}p_{x}$ makes this
$>\int_{0}^{\infty}e^{-rt}{}_{\frac{1}{r}}p_{x}(1-rt)\,dt$, which $=0$ by
calculus.
### 5.7. Life Annuity Duration
In the case of a bond or a LOIA, Macaulay duration agrees with life annuity
duration $Dr[a]$ as defined in Section 5.6. As above, for a CRIA, the
actuarial present value of payments is
$M(r,{\widehat{a}^{\star}})={\widehat{a}^{\star}}+F({\widehat{a}^{\star}},r)-G({\widehat{a}^{\star}},r)$.
Therefore with
$Dr[\widehat{a}^{\star}]=-\frac{1}{{\widehat{a}^{\star}}}\frac{d{\widehat{a}^{\star}}}{dr}$,
and Macaulay duration as defined as above, it follows from the formulas of
Section 5.4 that
$\text{Life Annuity Duration}=\frac{\text{Macaulay
Duration}}{(1+\pi)\frac{\partial(\frac{\pi}{1+\pi}{\widehat{a}^{\star}}+F-G)}{\partial{\widehat{a}^{\star}}}}.$
One can easily see the relationship between the two measure and they are
clearly not equal. Consider now the asymptotics of $Dr[\widehat{a}^{\star}]$
as $x$ rises to the maximal feasible age. ${\widehat{a}^{\star}}$ is the value
of $\alpha$ at which the convex function $F(\alpha)-G(\alpha)$ equals the
function $-\frac{\pi}{1+\pi}\alpha$. If that crossing takes place at very
large $\alpha$, that forces the graphs of the two functions to run nearly
parallel. At the limiting value of $x$, the convex curve will in fact be
asymptotic to the line. In other words, as $x$ approach this value,
$\lim{\alpha\to\infty}\frac{\partial(F-G)}{\partial\alpha}\uparrow-\frac{\pi}{1+\pi}$.
Therefore the denominator in the fraction displayed above will $\downarrow 0$,
so $Dr[\widehat{a}^{\star}]\to\infty$. That is,
$\frac{d{\widehat{a}^{\star}}}{dr}$ blows up even faster than
${\widehat{a}^{\star}}$ does.
### 5.8. Asymptotics for exponential mortality
An example where explicit calculations are possible is that of exponential
mortality, at rate $\lambda$. Here are the asymptotics of prices and life
annuity durations as $r\downarrow 0$, in the unloaded case. For CRIA and IRIA
we give two terms in the expansion.
* •
LOIA: price $=\frac{1}{r+\lambda}\to\frac{1}{\lambda}$ and life annuity
duration $=\frac{1}{r+\lambda}\to\frac{1}{\lambda}$.
* •
CRIA: price
$\approx\frac{1}{\lambda}\Big{[}\log(\frac{1}{r})-\log\log(\frac{1}{r})\Big{]}$
and life annuity duration
$\approx\frac{1}{r\log(\frac{1}{r})}\Big{[}1+\frac{\log\log(\frac{1}{r})}{\log(\frac{1}{r})}\Big{]}$
* •
IRIA: price
$\approx\frac{1}{\lambda}\Big{[}\log(\frac{1}{r})-2\log\log(\frac{1}{r})\Big{]}$
and life annuity duration
$\approx\frac{1}{r\log(\frac{1}{r})}\Big{[}1+2\frac{\log\log(\frac{1}{r})}{\log(\frac{1}{r})}\Big{]}$
We see that the LOIA price stays bounded while the IRIA and CRIA prices blow
up, but with the CRIA price higher. But the IRIA duration is the highest.
Here are the asymptotics of prices and life annuity durations as $r\to\infty$.
Again, we give two terms in the expansion.
* •
LOIA: price
$=\frac{1}{r+\lambda}\approx\frac{1}{r}\Big{[}1-\frac{\lambda}{r}\Big{]}$ and
life annuity duration
$=\frac{1}{r+\lambda}\approx\frac{1}{r}\Big{[}1-\frac{\lambda}{r}\Big{]}$.
* •
CRIA: price
$\approx\frac{1}{r}\Big{[}1-\frac{\lambda}{r}(1-\frac{1}{e})\Big{]}$ and life
annuity duration
$\approx\frac{1}{r}\Big{[}1-\frac{\lambda}{r}(1-\frac{1}{e})\Big{]}$
* •
IRIA: price $\approx\frac{1}{r}\Big{[}1-\frac{\lambda}{r}(\frac{2}{e})\Big{]}$
and life annuity duration
$\approx\frac{1}{r}\Big{[}1-\frac{\lambda}{r}(\frac{2}{e})\Big{]}$
Since $1>\frac{2}{e}>1-\frac{1}{e}$, here the CRIA values $>$ the IRIA values
$>$ the LOIA values.
## 6\. Conclusion
Although income annuities are the least popular of all annuities sold in the
U.S., comprising less than $5\%$ of all annuities sold in the U.S. during the
first quarter of 2021 according to a survey by LIMRA, they continue to be the
focus of much academic and scholarly interest. And, as noted in the literature
review (Section 2), most of the research on income annuities tends to focus on
contracts with life only features, or those with fixed guarantee periods,
whereas the majority of sales include a cash refund and instalment refund
feature. Moreover, as noted in the introduction, in late September 2021
approximately 77 million Americans with a defined contribution plan will come
face-to-face with annuity illustrations on their statements.
This is more than just an empirical observation about the type of insurance
riders that are preferred by consumers. Although motivated by these empirical
choices the main message of this paper is a theoretical one, namely that cash
refund income annuities lead to some very interesting and complex valuation
problems. As proved in Section 5, if valuation rates sink below a certain
threshold the CRIA is no longer viable and can no longer be offered. This
might help explain why inflation-linked income annuities no longer exist in
the U.S. market place; the term structure of real rates have likely declined
under that critical threshold.
There are a number of possible avenues for further research that follow from
the insights provided by this introductory article. First, it would be very
interesting to dig deeper into the money’s worth ratio (MWR) of cash-refund
income annuities (IA) to see whether their values are consistently higher that
the MWRs of life-only and period certain IAs. It’s quite possible CRIA reduces
the extent of anti-selection and that MWR values are higher for the consumer.
Table 6 provides preliminary evidence of that.
Second, and as a follow-up, perhaps CRIAs and their embedded life insurance
provide an “optimized” way of fulfilling a bequest motive, which might help
explain the relative demand for cash refunds over life only riders. Exploring
how anti-selection and mortality heterogeneity affects the valuation and
pricing of cash refund income annuities, is yet another avenue worth pursuing.
While there is a large body of literature focused on optimal life-only
annuitization strategies, it would be interesting to explore how those result
change once a cash-refund product is introduced and competes with a life only
version.
Third and finally, this entire paper has been predicated on the law of large
numbers that assumes the insurance company issuing the IA is selling enough so
that mortality risk is entirely diversifiable and pricing is done by (risk
neutral) expectations. In reality of course annuitant pools are of a finite
size, especially as the overall demand for income annuities is rather thin.
Just as importantly, mortality rates are not deterministic and Covid-19 was
just one data point that supports a stochastic mortality perspective. It would
be interesting to measure the extent to which ruin probabilities and the
corresponding amount of capital required is higher for CRIA. In time-honoured
tradition we leave these for subsequent research.
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## 7\. Appendix: R-script for computing CRIA values
The following R-script uses the bisection method (with 100 iterations)
together with R’s built-in integrate function to compute the value of a cash-
refund income annuity (CRIA) at age $x$, under a Gompertz law of mortality
with parameters $(m,b)$ and discount rate $r$. The integration scheme is based
on the representation provided in equation (5) in the body of the paper. In
Section 5 we used functions $F$ and $G$ to represent the left and right-hand
side. The resulting annuity price ${a^{\star}}$ would be accurate to within $1
per million premium.
CRIA<-function(x,r,m,b){ a_left<-0; a_right<-1/r for (i in 1:100){
a<-(a_left+a_right)/2
FINT<-function(t){exp(-r*t)*exp(exp((x-m)/b)*(1-exp(t/b)))}
GINT<-function(t){exp(-r*t)*r*(a-t)*exp(exp((x-m)/b)*(1-exp(t/b)))}
f<-integrate(FINT,a,Inf)$value-integrate(GINT,0,a)$value if (abs(f)<0.000001)
{break} if (f<0){a_right<-a} else {a_left<-a} } a } For example, assuming
$(x=65,m=90,b=10)$, the annual income generated by a premium of
$1,\\!000,\\!000$ in a CRIA, with discount rates ranging from $r=1\%$ to
$r=4\%$ would range from $51,164 to $67,103 per year (paid in continuous time)
and computed as:
> 1000000/CRIA(65,0.02,90,10) [1] 51164.71 > 1000000/CRIA(65,0.03,90,10) [1]
> 59169.89 > 1000000/CRIA(65,0.04,90,10) [1] 67103.97 For comparison, the
> life-only version at $r=2\%$, would generate $7,508 more income.
> x<-65; m<-90; b<-10; r<-0.02 >
> FINT<-function(t){exp(-r*t)*exp(exp((x-m)/b)*(1-exp(t/b)))} >
> a<-integrate(FINT,0,Inf)$value > 1000000/a [1] 58672.44
Type of income Annuity | Q1.2021 | Q4.2011
---|---|---
Life Only (with no guarantee) | 10.6% | 25.3%
Life with Period Certain | 30.0% | 56.2%
Refundable: Cash or Instalment | 59.4% | 18.5%
TOTAL: | 100% | 100%
Table 1. Market quotes for Income Annuities (IA) in the U.S., compiled
directly by the authors based on aggregate data provided by CANNEX Financial
Exchanges from their quarterly distributor activity survey experience. Note
over the last decade the sharp increase in demand for cash refund and
instalment refund IAs, and corresponding decline in life only IAs.
Age $(x)$ | $r=2\%$ | $r=4\%$
---|---|---
55 | 22.12615 | 16.82003
65 | 17.04378 | 13.73359
75 | 11.91615 | 10.17229
Table 2. Life Only Income Annuity (LOIA) prices $a$, with Gompertz mortality ($m=90,b=10$). For example under a $2\%$ valuation rate with no loadings, a $100,000 premium paid at the age of 65 would generate an income of $\$5,867.24=\frac{100000}{17.04378}$ per year, but would terminate upon death of the annuitant. Age $(x)$ | $r=2\%$ | $r=4\%$
---|---|---
55 | 23.79569 | 17.47113
65 | 19.54472 | 14.90225
75 | 15.19471 | 11.97156
Table 3. Cash Refund Income Annuity (CRIA) prices, with Gompertz mortality $(m=90,b=10)$. For example under a $2\%$ valuation rate with no loadings, a $100,000 premium at age 65 would generate an income of $\$5,116.47=\frac{100000}{19.54472}$ per year, but upon death of the annuitant the beneficiary would receive: $\$100,000$ minus the cumulative income received, if positive. Age $(x)$ | $r=2\%$ | $r=4\%$
---|---|---
55 | 23.55514 | 17.34376
65 | 19.18235 | 14.68173
75 | 14.7048 | 11.63911
Table 4. Instalment Refund Income Annuity (IRIA) prices, with Gompertz
mortality ($m=90,b=10$). For example under a $2\%$ valuation rate with no
loadings a $100,000 premium at age 65 would generate
$\$5,213.13=\frac{100000}{19.18235}$ per year, which is slightly more than the
CRIA. Upon death of the annuitant the beneficiary would continue to receive
$\$5,213$ until the $\$100,000$ premium was returned, instead of a lump-sum.
Age | Insurance Loading:
---|---
$(x)$ | $\pi=5\%$ | $\pi=15\%$ | $\pi=25\%$
55 | 16 | 46 | 74
65 | 23 | 65 | 105
75 | 35 | 101 | 163
Table 5. The lowest viable valuation rate denoted by $r_{\pi,x}$, expressed in basis points, under which the Cash Refund Income Annuity (CRIA) is still feasible with Gompertz Mortality ($m=90$ and $b=10$). Thus, for example, an inflation-adjusted CRIA would not be feasible at $x=75$, under a loading of $\pi=15\%$, if real rates used for pricing fall under $r_{0.15,75}=1.01\%$. Age & | Real & Live Quotes for $100,000 in IA Premium
---|---
Gender | Life Only IA | $\rightarrow$ MWR | Cash Refund IA | $\rightarrow$ MWR
65 M | $5,844 | 0.996 | $5,280 | 1.031
65 F | $5,556 | 1.005 | $5,112 | 1.043
80 M | $10,524 | 1.002 | $7,788 | 1.017
80 F | $9,636 | 1.008 | $7,428 | 1.033
Table 6. The Money’s Worth Ratio is computed using an $r=2\%$ valuation rate,
under Gompertz mortality with $m=90$ for males, $m=92$ for females and $b=10$
years for both. Data source is the website of Fidelity Investments on 10 July
2021, using non-qualified funds. Note MWR is higher for the cash-refund IA
when the same mortality assumptions are used.
Figure 1. Income Annuity (IA) prices (unloaded) under a variety of valuation
rates, assuming Gompertz mortality $(m=90,b=10)$ at age $x=65$ (left panel)
and $x=75$ (right panel). At relatively high (historical) valuation rates
there is little difference in prices. The refund has little impact. But at
(current) low $r$, the difference can be substantial.
Figure 2. Income Annuity (IA) prices under a valuation rate: $r=2\%$, with
Gompertz mortality $(m=90,b=10)$, comparing no loading with an insurance
loading of $\pi=15\%$. Notice how the loaded CRIA price increases beyond
(roughly) the age of $x=79$, and is no longer viable by (roughly) age $x=81$,
in the above figure.
Figure 3. Sensitivity of Income Annuity prices to age $x$, technically
$\frac{\partial}{\partial x}$, under a valuation rate of $r=2\%$ and Gompertz
mortality $(m=90,b=10)$. The partial derivatives of unloaded IA prices are all
negative since those pricess uniformly decline in age, but for the loaded
$\pi=15\%$ CRIA prices, the numbers can be positive; aging increases the
annuity price!
Figure 4. As defined and explained in the paper, life annuity durations at
various issue ages, under a valuation rate of $r=2\%$, and Gompertz mortality
$(m=90,b=10)$. The sensitivity of the unloaded actuarial price to interest
rates is (much) higher for the cash-refund and instalment refund IA,
especially at higher ages. For the loaded ($\pi=15\%$) IA, the situation is
reversed at younger ages, and the life annuity duration of the cash-refund
price “blows up” as $x$ reaches the critical age beyond 80.
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Annotating RNA motifs in sequences and alignments
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Preprint
Bio
Annotating RNA motifs in sequences and alignments
Paul P. Gardner1,2 *, Hisham Eldai1
Abstract
RNA performs a diverse array of important functions across all cellular life. These functions include important roles in
translation, building translational machinery and maturing messenger RNA. More recent discoveries include the miRNAs
and bacterial sRNAs that regulate gene expression, the thermosensors, riboswitches and other cis-regulatory elements
that help prokaryotes sense their environment and eukaryotic piRNAs that suppress transposition. However, there can
be a long period between the initial discovery of a RNA and determining its function. We present a bioinformatic
approach to characterise RNA motifs, which are the central building blocks of RNA structure. These motifs can, in
some instances, provide researchers with functional hypotheses for uncharacterised RNAs. Moreover, we introduce a
new profile-based database of RNA motifs - RMfam - and illustrate its application for investigating the evolution and
functional characterisation of RNA.
All the data and scripts associated with this work is available from: https://github.com/ppgardne/RMfam
Keywords
RNA — Motifs — Homology
1 School
of Biological Sciences, University of Canterbury, Christchurch, New Zealand.
Interaction Centre, University of Canterbury, Christchurch, New Zealand.
*Corresponding author: paul.gardner@canterbury.ac.nz
2 Biomolecular
1. Introduction
Characterising functional RNAs is an extraordinarily difficult
task. Even highly transcribed RNAs from model organisms
have remained uncharacterised for decades after their discovery. A specific example is the 6S sRNA, which was discovered
in 1971. The 6S sRNA is conserved across Bacteria and is
highly expressed in stationary-phase cells [1, 2]. But the role
of 6S as a regulator of RNA polymerase remained an enigma
for almost three decades [3]. Likewise, Y RNA, which was
discovered in 1981, is broadly conserved across metazoans
and is highly expressed [4]. It took two and a half decades
before Y RNAs were shown to be essential for the initiation
of DNA replication [5]. However, the mechanism for Y RNA
function still remains unclear. These and similar examples
show that it is remarkably difficult to functionally characterise
RNAs, even after decades of work.
A new generation of tools for RNA discovery is now available thanks to powerful new sequencing technologies. Entire
transcriptomes from species at different life stages, tissue
types and conditions can be studied with RNA-seq [6, 7, 8].
The total complement of RNA structures encoded in transcriptomes is also accessible with SHAPE-seq [9] and functional
regions of entire genomes of bacteria can be probed with techniques like TraDIS and Tn-seq [10, 11]. The data obtained by
these tools are unearthing novel RNAs at an unprecedented
rate, many of which are evolutionarily conserved, highly expressed, activated under specific conditions, essential and fold
into conserved secondary structures. Annotation efforts such
as those by the Rfam consortium [12, 13] are useful. However,
many RNAs are not found in this database and many that have
been curated remain uncharacterised [8]. To make sense of
the volumes of transcriptome data that is now being generated,
annotating this data and functionally characterising the cohort
of RNAs of Unknown Function (RUFs) is critical. A complication for such work is that evolutionary turnover, as well
as sequence variation can be high for ncRNAs [14, 15]. Consequently homology searches and other sequence-alignment
based analyses can be very challenging.
Many RNAs contain functional structures that recur both
within and across different RNA families. These motifs provide signatures that can identify functional components of
RNA sequences. The motifs that have been characterised to
date are involved in a diverse number of functions. These
include imparting structural stability, facilitating interactions
with other biomolecules, specifying cellular localisation and
coordinating gene regulatory signals [16, 17, 18, 19]
A number of publications detail bioinformatic methods
for the de novo discovery of RNA motifs from RNA primary sequences [20, 21]. There are also tools that can screen
RNA secondary structures [22] and RNA tertiary structures
[23]. The de scito (knowledge-based) approaches for the
annotation of RNA motifs include sequence and structure
descriptors [24, 25], primary and secondary structure-based
profile methods for specific motifs [26, 27] and even methods that combine primary, secondary and tertiary data [19].
We complement these approaches by introducing a resource
that identifies a range of previously characterised RNA motifs
in RNA sequences and alignments using covariance models
(CMs) [28, 29, 30, 31, 32].
Annotating RNA motifs in sequences and alignments — 2/9
We present 34 alignments, consensus structures and corresponding probabilistic models of published RNA motifs.
We call this resource RMfam, or RNA Motif Families (all
associated data and computer code is freely available from
our repository hosted on GitHub: http://github.com/
ppgardne/RMfam). These have been used to predict approximately 1, 900 conserved motifs in the Rfam (v11.0) alignments of RNA families; many of which are confirmed in the
published literature. Finally, we show examples of the applicability of our approach for studying RNA function, evolution
and alignment curation.
2. MATERIALS & METHODS
2.1 Distinction between Rfam and RMfam
The Rfam database collects and curates “seed alignments” of
RNA families. These are non-coding RNAs, cis-regulatory elements and self-splicing introns. The alignments are manually
constructed and annotated with consensus secondary structures, and used to seed probabilities for covariance models
(CMs) for each family. The Rfam CMs are widely used for
genome annotation projects to identify RNA loci (e.g. [33]).
A requirement before each family can pass Rfam qualitycontrol is that it is specific. In other words, there exists a
bit score threshold for each CM that distinguishes between
sequence matches that are related to the family and obvious
false-positive matches. Consequently, many RNA motifs are
not included in Rfam as they lack the required specificity
[34, 35, 36, 12, 13].
2.2 What is an RNA motif?
For the purposes of this work an RNA motif is a non-trivial,
recurring RNA sequence and/or secondary structure that can
be predominantly described by local sequence and secondary
structure elements. The motif is generally not restricted to
a particular family or taxonomic group. Note that in other
contexts, such as structural biology, a more general definition
of motif is frequently used, e.g. [37].
Accurate probabilistic methods for annotating structured
RNAs on DNA sequences called hidden Markov models
(HMMs) and covariance models (CMs) are now available
[28, 29, 30, 31, 32, 38]. From a given alignment, probabilistic models of conserved sequence (HMMs) and conserved
sequence plus secondary-structure (CMs) can be built and
used to filter large numbers of sequences for candidate homologous and/or analogous regions [39]. CMs cater to the
characteristics of RNA sequence evolution that are imposed
by basepairing (i.e. variation tends to preserve basepairing),
the result is that the accuracy of CMs is greater than alternative approaches [40]. The computational speed of CMs has
tended to be poor, however a lot of effort has been expended
on improving the speed of the approach while maintaining the
accuracy. Improvements include using HMMs as pre-filters
to accelerate CMs, query-dependent banding and Dirichlet
mixture priors [41, 39, 42, 38, 43].
RMfam sequences, structures and alignments were collated from a variety of heterogeneous and sometimes overlapping data repositories [12, 23, 44, 27, 45, 46, 47, 48, 37,
49, 50, 51]. Where possible we sourced data from publicly
accessible RNA motif resources, these included the FR3D
MotifLibrary [37], the models supplied with RMDetect [19],
the comparative RNA website [47] and SCOR [46]. We also
used information from specialised resources such as the kturn structural database [44] and SRPDB [52], as well as
generating our own alignments for motifs such as the ShineDalgarno and Rho-independent terminators based upon the
context of genome annotations (e.g. [27]). RNAFrabase was
frequently used as a source of RNA secondary structure annotations derived from PDB structures [53, 54]. Finally, where
necessary, we extracted sequences from publications. This
was often a manual effort, involving manually transcribing sequences and structures from figures in published manuscripts.
Where possible, these were mapped to PDB (downloaded
June 2014) nucleotide sequences [55, 56, 57], the EMBL
nucleotide archive [58] and Rfam (v11.0) [12, 13]. The provenance of each dataset is stored in the corresponding Stockholm
alignment. Each of these motifs were then passed through
quality control steps, where the sensitivity and specificity of
the resulting motif is assessed (See Figures 1 and S10-S43). If
these failed (e.g. the CM cannot identify member sequences
or the false-positive rate is extremely high), then the motif
was not included in the database. Each motif is also assigned
a curated score-threshold. This threshold (in bits) provides a
reasonable distinction between true and false matches.
2.3 Benchmarking motif annotations
In the following we briefly describe the benchmarks we have
used to evaluate our motif annotations. These are described
in further detail and with more elaborate results in the Supplementary Materials.
In order to determine the accuracy of our approach we
ran a series of three benchmarks. These were evaluated on
individual motifs (see Figures 1B and S10-S43), as well as
on the collective RMfam results (see Figures 1A and S9).
The first uses “RMfam sequences” which are taken from the
seed alignments. Ten shuffled sequences, with identical dinucleotide distributions, were generated for each RMfam seed
sequence [59]. Together these serve as positive and negative
controls for our test.
We constructed two further tests based upon Rfam (v11.0)
families. We identified Rfam families where there exists good
evidence (primarily based upon literature) that a motif is conserved in the family of related sequences (Supplementary
Table 1). These serve as positive controls for two further tests.
For the “Rfam sequences” benchmark we randomly selected at
Annotating RNA motifs in sequences and alignments — 3/9
least five sequences from each Rfam seed alignment (if fewer
than five sequences were available, then all were included).
We generated ten shuffled versions of each sequence; all had
an identical di-nucleotide distribution to the native sequence.
These sequences were all annotated with RMfam motifs, their
CM scores were recorded and used to evaluate the accuracy of
the annotations. Finally, for a “Rfam alignments” benchmark,
we evaluated the accuracy of RMfam annotations in an alignment context. Each Rfam alignment was filtered, removing
sequences more than 90% identical. The remaining sequences
were annotated with RMfam CMs, retaining only those that
cover more than 10% of the seed sequences and more than
two Rfam seed sequences. The summary statistic we use for
this final benchmark is a “sum-bits” score, this is the sum of
the bit scores for each match in all the sequences in a seed.
The accuracy metrics that we report here are the Matthew’s
correlation coefficient (MCC) [60], sensitivity and specificity.
The CMs built from RNA motifs tend to be short and
contain little sequence information. In RMfam the mean sequence length is just 34.3 nucleotides and the mean number of
basepairs is 10.9. Therefore scans of large sequence databases
with these models result in a number of false-positives. We
propose that annotating sequence alignments of ncRNAs have
the potential to improve the specificity of our predictions. This
assumes that evolutionarily conserved motifs are more likely
to be correct. In theory this approach could be extended to
genome alignments of e.g. transcribed regions.
3. RESULTS
In this study we present 34 RMfam alignments and probabilistic models of published RNA motifs (all freely available from
our repository hosted on GitHub: http://github.com/
ppgardne/RMfam). These have been used to predict approximately 2, 500 conserved motifs in the Rfam (v11.0) seed
alignments; many of which are confirmed in the published
literature. Furthermore, our permutation tests have shown
that both the sensitivity and specificity of this approach is
remarkably high given the short motifs we use (See Figures 1
and S9-S44).
3.1 Function
One of the most labour intensive stages of RNA research is
identifying the function of newly discovered RNAs. In order to illustrate the utility of RMfam for this task we show
the matches between a model of the CsrA-binding site and
two RNA families of unknown function, TwoAYGGAY and
Bacillaceae-1 (Rfam IDs RF01731 and RF01690, see Figure 2). CsrA is a bacterial RNA binding protein that regulates
the translation and stability of mRNAs [62]. It binds mRNAs
carrying CsrA binding motifs, physically occluding ribosomebinding sites. This binding can itself be regulated by competition between the mRNAs and highly expressed sRNAs
that host numerous CsrA binding sites. However, this class of
sRNA (CsrB, CsrC, RsmX, RsmY and RsmZ) has only been
identified in Gammaproteobacteria [63, 64]. The ncRNAs,
TwoAYGGAY and Bacillaceae-1, were initially discovered in
a large-scale bioinformatic screen [65]. Some further analysis
identified two tandem-GAs in one of the stems that characterise the structure of TwoAYGGAY [19]. The matches
between these families and the CsrA binding motif were discovered in this work and provide a testable hypothesis for
further validation that there are also CsrA binding sRNAs in
Clostridia (TwoAYGGAY), and Bacillales and Lactobacillales
(Bacillaceae-1). The validation of these predictions is a work
in progress with our collaborators.
3.2 Evolution
Non-coding RNAs are remarkably tolerant of genetic variation, as evident by the wide degree of sequence variation
that can be found between evolutionarily related ncRNAs
[66, 67, 68, 15]. However, structure frequently constrains the
evolution of RNA sequences. That said, structures can also be
dynamic. For example, motifs that confer structural stability
can be exchanged over time, resulting in a rich and complex
evolutionary history. This illustrates that studying the gain
and loss of RNA motifs over evolutionary time-scales can
help characterise the dynamic evolution of RNA sequences
and structures.
A good example of this is the Lysine riboswitch. This is a
convenient example, that for illustrative purposes that we will
describe in further detail. As illustrated in Figure 3 many motifs may be exchanged, e.g. the U-turn motif with a k-turn in
the P2 stem or the T-loop and the GNRA tetraloop in stem P4.
Interestingly, the motif distributions are relatively clade-like,
with closely related riboswitches more likely to share motifs, e.g. the GNRA tetraloop found in the Pasteurellales and
Vibrionales taxonomic groups. This type of annotation information is valuable for researchers investigating the structure
and evolution of RNA families.
3.3 Curation
Another use of the results presented in this work is of importance for the curators of RNA alignments and sequences
[12, 69, 70]. Until now it has been difficult to analyse the evolutionary conservation of motifs in the context of an alignment,
although some progress has been made when crystallographic
data is available, e.g. the RNASTAR collection of structural
RNA alignments [70]. With the help of RMfam, malformed
alignments can be detected and corrected where conserved
RNA motifs are incorrectly aligned. We illustrate an example
of this for the Rfam (v11.0) 5S rRNA alignment that contains
a misaligned, yet highly conserved sarcin-ricin motif (see Figure S45), and for the Rfam RsmY alignment, which is a CsrA
binding sRNA. The RsmY alignment has a mis-annotated
consensus structure that does not include a further CsrA binding motif (see Figure S46). These motifs generally occur in
pairs, as CsrA is a homodimeric protein, with each half of the
protein binding a motif [71, 72].
Annotating RNA motifs in sequences and alignments — 4/9
B.MCC values
x
0.6
sum−bits=145
maxMCC=0.68
x
x
0.4
bits=18.5
maxMCC=0.66
0.2
Sensitivity
0.8
1.0
A. ROC plot
0.0
Rfam alignments (sum−bits)
Rfam sequences (bits)
RMfam sequences (bits)
0.0
0.2
0.4
0.6
bits=19
maxMCC=0.38
0.8
Specificity
x
1.0
Rfam alignments (sum−bits)
Rfam sequences (bits)
RMfam sequences (bits)
HuR_binding
VTS1_binding
vapC_target
C−loop
CUYG
k−turn−1
AUF1_binding
UAA_GAN
RBS_E_coli
Roquin_binding
RBS_H_pylori
ANYA
RBS_B_subtilis
TRIT
sarcin−ricin−1
right_angle−3
T−loop
GNRA
UNCG
U−turn
UMAC
sarcin−ricin−2
CsrA_binding
right_angle−2
docking_elbow
tandem−GA
SRP_S_domain
k−turn−2
Terminator2
CRC_binding
Terminator1
pK−turn
twist_up
Domain−V
0.0
1.0
2.0
3.0
max(MCC)
Figure 1. In the above plots we assess the accuracy of motif annotation and test whether annotating alignments instead of
sequences improves the prediction accuracy. We have applied three different benchmarks (described in the text). In sub-figure
A we show a ROC plot for pooled RMfam annotations. This plots the sensitivity versus specificity of all the motif annotations
on sequences or alignments at different score thresholds. The ’x’s illustrate where on the curve the maximum Matthew’s
Correlation Coefficient is located, and the corresponding bit scores are indicated. In sub-figure B we illustrate the maximum
MCC of CM annotation for each motif from the 3 different benchmarks.
4. DISCUSSION & CONCLUSION
The chief motivation for this work is to functionally characterise novel ncRNAs. Our vision for the RMfam resource is to
annotate RNAs of unknown function (e.g. [8]). These motif
annotations will help develop further functional hypotheses
and accelerate experimental characterisation.
In this work, we have shown that RMfam is surprisingly
accurate. Despite the fact that the average RMfam motif consists of just 34.3 nucleotides and 10.9 basepairs, we show that
the covariance models are specific enough to distinguish between motif-hosting sequences and negative control sequences
(See Figures 1 and S10-S43). Our approach shows improved
performance when evolutionary information encoded in Rfam
sequence alignments is incorporated into the predictions. We
hypothesise that annotated genome alignments may be a useful source of motifs and we will investigate this idea further
in future. As a discovery tool this resource has already made
some useful predictions. We have predicted the existence
of two new CsrA binding ncRNAs, potentially the first of
this class of regulatory molecules to be found outside of the
Gammaproteobacteria. However, further work needs to be
carried out to validate this claim.
4.1 Future work and potential applications
We have identified some future developments and applications
for the RMfam resource. We plan to continue developing the
accuracy of the motif annotation tools as well as increase the
access to RMfam annotations via other databases and expand
the number of motifs included in RMfam. Furthermore, it may
be possible to boost the accuracy of RNA secondary structure
prediction tools by constraining these with predicted motifs.
We elaborate further on these ideas below.
The Lysine riboswitch example raises the possibility that
certain types of motif are preferentially exchanged during
the evolution of ncRNAs. Do stable hairpin motifs such as
the GNRA and T-loops replace each other more frequently
than we expect by chance? This would blur the lines between
our understanding of homologous and analogous structures
[73]. Another possibility is that certain motifs co-occur more
frequently than we expect. For example, are k-turns more
frequently closed by U-turns than we expect? If correct, these
enrichments of favoured exchanges and co-occurances could
be used to increase our confidence in motif annotations and
Annotating RNA motifs in sequences and alignments — 5/9
Y GG
A
A
C
Y
C G
G G
C
G
G
Y
Y
Y
R
U
GG A
A
C G
5´
AY
G
GY Y
G
G
Y AC R R
RC
R Y
CsrA binding motif
5'
R
Y R
G
G R
R C RA
C
G
YGY
G
U
A
R Y
G C
CA
GUY
Core TwoAYGGAY
GG A
U
A
C G
R Y
Y R
C G
Y R
5'
U
GG A
A
C
C G
A U
C G
C G
G YG
Y
U
U
C
G
U
C GAU
G C
G C
Y R
R Y
Y G
A
Y UCCUUY
Legend
base pair annotations
covarying mutations
compatible mutations
no mutations observed
YY
RevComp Bacillaceae-1
nucleotide
nucleotide
present
identity
80% 60% N 80%
70% 40% N 70%
N 60%
R = A or G. Y = C or U.
Figure 2. The secondary structures and sequence conservation of CsrA binding motif and two new candidate CsrA binding
sRNAs, TwoAYGGAY and Bacillaceae-1 illustrated with R2R [61]. These families each have two strong matches to the
CsrA-binding motif, this new evidence provides a strong case that these RNAs regulate the activity of the regulatory protein,
CsrA, by sequestering this nucleotide-binding protein. The “core” of the TwoAYGGAY structure is shown, the Rfam (v11.0)
model contains a further external stem that is not well conserved. Also, the reverse-complement (RevComp) of the
Bacillaceae-1 is illustrated, this strand has the matches to the CsrA binding motif and the original discoverers of this ncRNA
are not confident of the strand (personal communication, Weinberg Z).
can assist with the design of functional RNAs.
Typical RNA structure prediction methods to not incorporate information about RNA motifs. We propose that RMfam predictions can be used as constraints for existing RNA
structure prediction software, thus improving the accuracy of
structure prediction tools which can often be inaccurate [74].
This approach is analogous to the fragment-library approach
that is frequently used for tertiary structure prediction [75].
Another application for RMfam covariance models is as a
pre-filter to accelerate the more complex methods, for example, the Bayesian network approach implemented in RMdetect
[19].
Increasing the access of motif annotations is another goal
of the authors. We are active in the Rfam and RNAcentral
consortia, both of which curate non-coding RNAs, the former
ncRNA alignments and the latter ncRNA sequences [12, 69,
13]. Our results show that curators can benefit greatly from
motif annotations (see Figures S44-S45) and it is likely that
RMfam annotations will be incorporated into these databases
in future releases.
New technologies such as the sequencing of cross-linked
RNA and protein are a potential source of new RNA-protein
motifs. In the future we will mine these datasets [76, 77,
78] for new additions to the RMfam database. Furthermore,
we will continue to add new motifs to RMfam as they are
published.
Finally, as previously mentioned, the specificity of the
RMfam annotations is generally low. However, incorporating
the genomic and taxonomic context of annotations into the
predictions may result in performance gains. For example,
Shine-Dalgarno and rho-independent terminators are generally located in bacterial sequences and at the extremities of
annotated genes. A probabilistic incorporation of contextual
information will likely result in further performance gains.
In summary, we have developed a resource for annotating
diverse sets of RNA motifs in nucleotide sequences and alignments. We have proven the accuracy using benchmarks, and
the utility of this resource for alignment curation, evolutionary
analyses and shown that it has some promise for the prediction
of RNA function.
5. ACKNOWLEDGEMENTS
This work would not be possible without input from a large
community of RNA researchers that openly share their results. It has benefited from many discussions with members
of the Xfam Consortium, the RNA Ontology Consortium and
attendees of the 2012 Benasque Meeting on RNA. A special
thanks to Lars Barquist, Elena Rivas, Rob Knight, Eric Westhof, Zasha Weinberg, Anthony Poole, Peter Fineran and two
anonymous reviewers for their valuable contributions.
PPG is supported by a Rutherford Discovery Fellowship
from Government funding, administered by the Royal Society
of New Zealand.
5.0.1 Conflict of interest statement.
None declared.
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AMPH1 functions as a tumour suppressor in ovarian cancer via the inactivation of PI3K/AKT pathway
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Yajun Chen
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English
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Spoken
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Funding information Funding information
Shanghai Collaborative Innovation Center
for Translational Medicine, Grant/Award
Number: TM201923; Shanghai Municipal
Key Clinical Specialty, Grant/Award
Number: shslczdzk06302; National Natural
Science Foundation of China, Grant/Award
Number: 81572547 AKT pathway, AMPH1, anti-oncogene, ovarian cancer, PI3K, tumour suppressor O R I G I N A L A R T I C L E O R I G I N A L A R T I C L E Abstract Abstract
AMPH1, an abundant protein in nerve terminals, plays a critical role in the recruit-
ment of dynamin to sites of clathrin-mediated endocytosis. Recently, it is reported
to be involved in breast cancer and lung cancer. However, the impact of AMPH1 on
ovarian cancer is unclear. In this study, we used gain-of-function and loss-of-function
methods to explore the role of AMPH1 in ovarian cancer cells. AMPH1 inhibited
ovarian cancer cell growth and cell migration, and promoted caspase-3 activity, re-
sulting in the increase of cell apoptosis. In xenograft mice model, AMPH1 prevented
tumour progression. The anti-oncogene effects of AMPH1 on ovarian cancer might
be partially due to the inhibition of PI3K/AKT signalling pathway after overexpres-
sion of AMPH1. Immunohistochemistry analysis showed that the staining of AMPH1
was remarkably reduced in ovarian cancer tissues compared with normal ovarian tis-
sues. In conclusion, our study identifies AMPH1 as a tumour suppressor in ovarian
cancer in vitro and in vivo. This is the first evidence that AMPH1 inhibited cell growth
and migration, and induced apoptosis via the inactivation of PI3K/AKT signalling
pathway on ovarian cancer, which may be used as an effective strategy. 2The international Peace Maternity and
Child Health Hospital, School of Medicine,
The China Welfare Institute, Shanghai Jiao
Tong University, Shanghai, China
3Shanghai Key Laboratory of Embryo
Original Diseases, Shanghai, China
4Shanghai Municipal Key Clinical Specialty,
Shanghai, China Correspondence
Tianying Zhong and Lihua Wang,
Department of Clinical Laboratory, Nanjing
Maternity and Child Health Care Hospital,
Women’s Hospital of Nanjing Medical
University, Nanjing 210004, China. Emails: tying_zhong22@163.com;
2628451306@qq.com Received: 17 December 2019 | Revised: 4 April 2020 | Accepted: 27 April 2020 Received: 17 December 2019 | Revised: 4 April 2020 | Accepted: 27 April 2020 Received: 17 December 2019 | Revised: 4 April 2020 | Accepted: 27 April 2020
DOI: 10.1111/jcmm.15400 Yajun Chen1 | Wenjiao Cao2,3,4 | Lihua Wang2,3,4 | Tianying Zhong1 Yajun Chen1 | Wenjiao Cao2,3,4 | Lihua Wang2,3,4 | Tianying Zhong1 Yajun Chen1 | Wenjiao Cao2,3,4 1Department of Clinical Laboratory, Nanjing
Maternity and Child Health Care Hospital,
Women’s Hospital of Nanjing Medical
University, Nanjing, China
2The international Peace Maternity and
Child Health Hospital, School of Medicine,
The China Welfare Institute, Shanghai Jiao
Tong University, Shanghai, China
3Shanghai Key Laboratory of Embryo
Original Diseases, Shanghai, China
4Shanghai Municipal Key Clinical Specialty,
Shanghai, China J Cell Mol Med. 2020;24:7652–7659. 2.4 | Western blot assay Total proteins extracted from all cells were resolved on SDS-
PAGE. And then, proteins on gel were transferred to PVDF mem-
branes which were blocked with 5% BSA in TBST for 1 hour and
incubated with corresponding primary antibodies overnight at
4°C. AMPH-1, p-PI3K(Tyr458), P-AKT(Ser473), PI3K, AKT and
β-actin antibodies were purchased from Proteintech and Cell
Signaling. The membranes were incubated with secondary an-
tibody for 1 hour at room temperature. Finally, an enhanced
chemiluminescence system was further performed to detect
protein levels. Ovarian cancer, the seventh most commonly diagnosed carci-
noma in the world, is one of the most aggressive type's gynaecologic
cancers.12,13 Approximately 85% ovarian cancer cases are diagnosed
at an advanced stage (III/IV) due to the lack of screening for early
diagnosis of ovarian cancer.12 As an aggressive tumour, ovarian can-
cer is characterized by the tendency of metastasizing early.14 Recent
years effective treatment strategies have developed diversely; how-
ever, survival outcomes barely improved.15 Therefore, it is impera-
tive to identify novel therapeutic strategy for ovarian cancer. 2.3 | Cell transfection The stable cell lines with AMPH1 knockdown were gener-
ated by integration of lentiviral shRNA vectors specific for
AMPH-1. The shRNA against AMPH-1 shown as follows:
5′-GCGAGAACUCCGAGGAUAUTT-3′. The PCMV-AMPH overex-
pression plasmid (abnova) was used to construct AMPH1 overex-
pression cell lines. 2.5 | QRT-PCR Here, the roles of AMPH1 in ovarian cancer were systematically
investigated, and the mechanism underlying these effects was also
explored. This study demonstrated that AMPH1 functioned as a
tumour suppressor in ovarian cancer. Specifically, the anti-onco-
gene effect of AMPH1 may through induce apoptosis via promot-
ing caspase-3 activity and the inactivation of PI3K/AKT signalling
pathway. Total RNA was extracted by using TRIzol reagent (TaKaRa Bio). And then, RNA was reverse-transcribed into complementary DNA
(cDNA) by using a First-Strand cDNA Synthesis kit (TaKaRa Bio). SYBR mixed with cDNA was used. The relative expression level of
AMPH1 in each type of cell was analysed by using 2−ΔΔCt method. Primers were listed below. 2.2 | Cell culture AMPH can express in non-neuronal tissues. The 108 kD iso-
form of AMPH1 is the predominant isoform that expressed out-
side the brain in human, and it represents an alternatively spliced
variant of neuronal AMPH1 missing a 42 amino acid insert.5
AMPH1 is first identified as a human autoantigen in neurologi-
cal paraneoplastic autoimmune diseases with breast cancer,6 and
then AMPH1 antibodies are detected in the serum of patients
with small-cell lung cancer.7 Further study reported a link be-
tween AMPH1 expression in cancer and AMPH1 autoimmunity,
and this is the first proof suggesting AMPH family members may
be associated with cancer.5 Subsequently, Otsuka et al8 demon-
strated that AMPH1 inhibited the adhesion and spreading of
HeLa cells as well as migration of melanoma cells by binding to
vitronectin. In recent studies, the association between AMPH1
and tumour progression is well investigated. It is reported that
AMPH1 expressed lower in SKMES-1 and A549 cells.9 In addition,
AMPH1 can function as a tumour suppressor in breast cancer
cells. Chen et al10 found that inhibition of AMPH1 induced cell
proliferation, cell migration and cell cycle progression, but inhib-
ited cell apoptosis, which might be partially due to the activation
of EMT and ERK pathways. In lung cancer, Yang et al11 also iden-
tified the anti-oncogenic function of AMPH1 in vitro and in vivo,
and the suppression effects were mediated by Ras-Raf-MEK-ERK
signalling pathway. However, up to now, no study reports the in-
fluences of AMPH1 on ovarian cancer. Two human ovarian cancer cell lines, Caov-3 and Skov3 cells,
were obtained from American Type Culture Collection (ATCC,
USA). Cells were cultured in RPMI 1640 with 10% foetal bovine
serum (FBS, Gibco) at 37°C in a humidified incubator filled with
5% CO2. 1 | INTRODUCTION The recruitment mediates the last step in synaptic vesicle endocyto-
sis, an essential neurophysiologic component in synaptic transmis-
sion.4 AMPH1 also serves as a multifunctional adaptor connecting
clathrin coat proteins, dynamin, synaptojanin and lipid membranes.2
In the proline-rich domain, AMPH1 includes several phosphorylation
sites for protein kinases, like cyclin-dependent kinase 5, a member
of the cyclin-dependent protein kinase family, which plays a role in
neuronal migration and neurite outgrowth via its action on the actin
cytoskeleton.1,2 Amphiphysin 1 (AMPH1), a phosphoprotein expressed at high lev-
els in neurons, participates in synaptic vesicle endocytosis and neu-
rite outgrowth.1 It belongs to a protein family conserved from yeast
to humans, playing pleiotropic roles in actin function, and regulation
of growth control.1 AMPH1 interacts with other endocytic proteins,
including dynamin and adaptor Protein Complex 2, involved in the
recruitment of dynamin to sites of chathrin-mediated endocytosis.2,3 7652 |
J Cell Mol Med. 2020;24:7652–7659
wileyonlinelibrary.com/journal/jcmm
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium,
provided the original work is properly cited. © 2020 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd. Chen and Cao contributed equally to this work. ope y c ted. Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd. J Cell Mol Med. 2020;24:7652–7659. 7653 CHEN et al. 2.8 | Cell apoptosis assay Annexin-VFITC Apoptosis Detection Kit I (BD Biosciences) was
used for analysing apoptosis. cells were resuspended by cold
PBS at a density of 1106 cells/ml, and incubated with PI and
Annexin-VFITC staining, followed by incubation at room tem-
perature for 15 minutes. Flow cytometry was used to analyse
apoptosis. AMPH1 staining was evaluated by pathologists who were blinded
to the sample origins and the patient outcomes. The widely accepted
German semi-quantitative scoring system was used to score staining
intensity and extent in different areas. Each specimen was scored
according to the intensity of the nucleic, cytoplasmic and membrane
staining (weak staining = 1, moderate staining = 2 and strong stain-
ing = 3) and the proportion of stained cells (0%-5% = 0, 5%-25% = 1,
26%-50% = 2, 51%-75% = 3 and 76%-100% = 4). The final immunore-
activity score was determined by multiplying the intensity score by
the proportion score. All IHC scores were ranged from 0 (the mini-
mum score) to 12 (the maximum score). 2.1 | Tissue samples from patients with ovarian
cancer The Caspase-3 Activity Assay was done according to the protocol
of the Caspase-3 Activity Assay Kit (Cell Signaling, #5723). It con-
tains a fluorogenic substrate (N-Acetyl-Asp-Glu-Val-Asp-7-amino-
4-methylcoumarin or Ac-DEVD-AMC) for caspase-3. During the
assay, activated caspase-3 cleaves this substrate between DEVD
and AMC, generating highly fluorescent AMC that can be detected
using a fluorescence reader with excitation at 380 nm and emission
between 420 and 460 nm. A total 15 ovarian cancer specimens and 15 normal ovarian tis-
sues were obtained from Nanjing Maternity and Child Health Care
Hospital. All patients have written informed consent. Fresh speci-
mens were immediately fixed in formalin and embedded in wax for
immunohistochemistry. 7654 |
2.7 | Cell growth assay 7654 CHEN et al. cells was determined. The width ratio was calculated by the wound
width/the distance measured at 0 hour. cells was determined. The width ratio was calculated by the wound
width/the distance measured at 0 hour. Cell growth was detected via Cell Counting Kit-8 (CCK-8) assay. Cells
were transformed to 96-well plates at a density of 3000 cells/well. CCK-8 experiment was performed according to the manufacturer's
instructions. Absorbance at 490 nm was obtained by a microplate
reader (BioTek Instruments, Inc). OD values were determined after
24, 48, 72 and 96 hours. 2.10 Human tumour tissues embedded in paraffin were sliced into 5 μm
sections for staining. To retrieve antigenic activity, the deparaffi-
nized and rehydrated sections were heated in citrate buffer at 121°C
for 30 minutes. Blocking was done by incubation with 10% goat
serum at room temperature for 30 minutes. Sections were incubated
with AMPH1 antibody for overnight at 4°C, followed by staining
with secondary antibody for 1 hour at room temperature. Sections
were finally stained with 3, 3-diaminobenzidine tetrahydrochloride
and counterstained with haematoxylin. 2.12 | Statistical analysis Data are presented as mean ± SEM. All experiments were performed
at least three times. Data were assessed using the software SPSS
19.0. P < .05 was regarded statistically significant difference. 3.2 | AMPH1 significantly induced the cell
apoptosis of ovarian cell lines We next evaluated the effect of AMPH1 on apoptosis in Caov-3
and Skov3 cells. Flow cytometry results showed that the absence of
AMPH1 significantly prevented cell apoptosis, whereas the increase of
AMPH1 expression in ovarian cancer cell lines significantly induced cell
apoptosis (Figure 2A,B). One of the best-known markers of apoptosis
is the proteolytic cleavage of pro-caspase-3 into its active form.16,17 We next evaluated the effect of AMPH1 on apoptosis in Caov-3
and Skov3 cells. Flow cytometry results showed that the absence of
AMPH1 significantly prevented cell apoptosis, whereas the increase of
AMPH1 expression in ovarian cancer cell lines significantly induced cell
apoptosis (Figure 2A,B). One of the best-known markers of apoptosis
is the proteolytic cleavage of pro-caspase-3 into its active form.16,17
Caspase-3, a main executor of apoptosis, specifically cleaved substrates
and finally results in DNA fragmentation and cell apoptosis. To further
define the apoptosis involved by AMPH1, we detected the activity of
caspase-3 in Caov-3 and Skov3 cells. AMPH1 knockdown markedly 2.9 | Wound healing assay Cells were transformed to six-well plates with 10% FBS. When 80%
of the well was covered with cells, a 100 μl pipette tip was used
to scrap the cells. As a result, wounds were generated. Then, cells
were incubated at 37°C. After 24 hours, the migration distance of FI G U R E 1 AMPH1 significantly suppressed the cell proliferation of ovarian cancer cell lines. A, Western blot assay showed that the
protein level of AMPH1 in AMPH-1 silenced (ShAMPH1) or overexpressed (AMPH-OE) Caov-3 and Skov3 cells. B, qRT-PCR showed the
mRNA level of AMPH1 in ShAMPH1 or AMPH-OE Caov-3 and Skov3 cells. C, MTT assay showed that knockdown of AMPH1 significantly
promoted cell growth, and overexpression of AMPH1 suppressed the cell proliferation of ovarian cancer cell lines. * P < .05. ***P < .001 FI G U R E 1 AMPH1 significantly suppressed the cell proliferation of ovarian cancer cell lines. A, Western blot assay showed that the
protein level of AMPH1 in AMPH-1 silenced (ShAMPH1) or overexpressed (AMPH-OE) Caov-3 and Skov3 cells. B, qRT-PCR showed the
mRNA level of AMPH1 in ShAMPH1 or AMPH-OE Caov-3 and Skov3 cells. C, MTT assay showed that knockdown of AMPH1 significantly
promoted cell growth, and overexpression of AMPH1 suppressed the cell proliferation of ovarian cancer cell lines. * P < .05. ***P < .001 | 7655 7655 7655 CHEN et al. FI G U R E 2 AMPH1 induced cell apoptosis and promoted caspase-3 activity. A, Flow cytometry results showed that knockdown of
AMPH1 suppressed ovarian cancer cell apoptosis, whereas overexpression of AMPH1 stimulates cell apoptosis. B, The percentage of
apoptotic cells in each type of cell was calculated. C, Knockdown of AMPH1 inhibited caspase-3 activity, and overexpression of AMPH1
promoted caspase-3 activity. *P < .05. **P < .01. ***P < .001 FI G U R E 2 AMPH1 induced cell apoptosis and promoted caspase-3 activity. A, Flow cytometry results showed that knockdown of
AMPH1 suppressed ovarian cancer cell apoptosis, whereas overexpression of AMPH1 stimulates cell apoptosis. B, The percentage of
apoptotic cells in each type of cell was calculated. C, Knockdown of AMPH1 inhibited caspase-3 activity, and overexpression of AMPH1
promoted caspase-3 activity. *P < .05. **P < .01. ***P < .001 2.11 | Xenograft mice model the protein and mRNA levels of AMPH1 respectively after transfec-
tion. As shown in Figure 1A,B, AMPH1 protein and mRNA levels were
decreased in sh AMPH1 cells. Conversely, the protein and mRNA lev-
els increased in AMPH-OE cells (Figure 1A,B). Subsequently, we fur-
ther explored the ability of cell proliferation. MTT results showed that
knockdown of AMPH1 remarkably aggravated the accumulation of
cells compared with cells without transfection (Figure 1C). However,
overexpression of AMPH1 significantly inhibited the growth of
Caov-3 and Skov3 cells (Figure 1C). Female BALB/c nude mice aged 4- to 6-week old were used. Mice were
divided to three groups, and subcutaneously injected with ShAMPH1
or AMPH-OE Skov3 cells, respectively. After 21 days, mice were sacri-
ficed. Tumour volume and weight in each group were measured. 3.4 | AMPH1 inhibited the activation of PI3K/AKT
signalling pathway FI G U R E 4 AMPH1 inhibited the activation of PI3K/AKT
signalling pathway in ovarian cancer cells. Western blot assay
showed that knockdown of AMPH1 increased the protein levels of
p-PI3K and p-AKT, whereas overexpression of AMPH1 inhibited
these proteins levels. In addition, silencing AMPH1 inhibited the
expression of E-cadherin The PI3K/AKT signalling pathway is one of many mechanisms
that regulate cell cycle and cell apoptosis, and dysregulation of a
component in this pathway leads to cancer.18 Therefore, we fur-
ther detected the association between AMPH1 and PI3K/AKT
signalling pathway. Inhibiting AMPH1 expression induced the
accumulation of p-PI3K and p-AKT proteins, and AMPH1 over-
expression suppressed these proteins levels, revealing AMPH1
might regulate ovarian cancer progression via PI3K/AKT sig-
nalling pathway (Figure 4). Loss of E-cadherin gene expression
causes dysfunction of cell junction system, allowing cancer cell
invasion and metastasis. To elucidate the underlying mecha-
nism by which AMPH1 regulates cell invasion, we investigated
expression of E-cadherin in Caov-3 and Skov3 cells. Compared
with shN cells, silencing AMPH1 had a decreased expression of
E-cadherin at protein levels (Figure 4), suggesting AMPH1 regu-
lated the E-cadherin. suppressed caspase-3 activity, and AMPH1 overexpression markedly
promoted caspase-3 activity (Figure 3C), indicating AMPH1 was as a
tumour suppressor to promote ovarian cancer cell apoptosis. 3.1 | AMPH1 significantly suppressed the cell
proliferation of ovarian cancer cell lines To detect the roles of AMPH1 in ovarian cancer, gain-of-function and
loss-of-function methods were used. We generated the AMPH-1 si-
lenced (ShAMPH1) or overexpressed (AMPH-OE) stable Caov-3 and
Skov3 cells. Western blotting and qRT-PCR were performed to verify Caspase-3, a main executor of apoptosis, specifically cleaved substrates
and finally results in DNA fragmentation and cell apoptosis. To further
define the apoptosis involved by AMPH1, we detected the activity of
caspase-3 in Caov-3 and Skov3 cells. AMPH1 knockdown markedly 7656 | CHEN et al. CHEN et al. 7656 FI G U R E 3 AMPH1 significantly inhibited cell migration of ovarian cancer cell lines. A, Wounding healing assay showed that AMPH1
suppressed cell migration in Caov-3 and Skov3 cells. Migration of cells to the wound was visualized at 0 and 24 h with a microscope. B, The
width ratio was calculated by the wound width/the distance measured. **P < .01. ***P < .001 FI G U R E 3 AMPH1 significantly inhibited cell migration of ovarian cancer cell lines. A, Wounding healing assay showed that AMPH1
suppressed cell migration in Caov-3 and Skov3 cells. Migration of cells to the wound was visualized at 0 and 24 h with a microscope. B, The
width ratio was calculated by the wound width/the distance measured. **P < .01. ***P < .001 FI G U R E 4 AMPH1 inhibited the activation of PI3K/AKT
signalling pathway in ovarian cancer cells. Western blot assay
showed that knockdown of AMPH1 increased the protein levels of
p-PI3K and p-AKT, whereas overexpression of AMPH1 inhibited
these proteins levels. In addition, silencing AMPH1 inhibited the
expression of E-cadherin metastasizing early.13,14 Metastasis is generally connected with
poor prognosis. Thus, here, we evaluated the impact of AMPH1
on the cell migration in Caov-3 and Skov3 cells by using wound-
ing healing assay. Migration of cells to the wound was visualized
at 0 and 24 hours with a microscope (Figure 3A). The width ratio
was calculated by the wound width/the distance measured at
0 hour (Figure 3B). The results showed that the width ratio was
higher in ovarian cancer cells with sh AMPH1 compared with
control cells, and lower in cells with AMPH1 overexpression. These findings suggest that silence of AMPH1 induced cell mi-
gration; in turn, the accumulation of AMPH1 inhibited the ability
of cell migration. 3.3 | AMPH1 significantly inhibited cell migration of
ovarian cancer cell lines Ovarian cancer is one of the most aggressive type's gynaeco-
logic cancers in the world characterized by the tendency of 7657 CHEN et al. |
FI G U R E 5 AMPH1 inhibited tumour growth in vivo. A, The images of tumours injection with ShAMPH1 or AMPH-OE Skov3 cells. B,
Knockdown of AMPH1 increased tumour volume, whereas overexpression of AMPH1 decreased tumour volume. C, Knockdown of AMPH1
increased tumour weight, whereas overexpression of AMPH1 decreased tumour weight. **P < .01. ***P < .001. D, The protein expression of
AMPH1, p-PI3K, p-AKT and E-cadherin in xenograph tumours FI G U R E 5 AMPH1 inhibited tumour growth in vivo. A, The images of tumours injection with ShAMPH1 or AMPH-OE Skov3 cells. B,
Knockdown of AMPH1 increased tumour volume, whereas overexpression of AMPH1 decreased tumour volume. C, Knockdown of AMPH1
increased tumour weight, whereas overexpression of AMPH1 decreased tumour weight. **P < .01. ***P < .001. D, The protein expression of
AMPH1, p-PI3K, p-AKT and E-cadherin in xenograph tumours 3.5 | AMPH1 significantly inhibited ovarian tumour
progression in vivo to the intensity of the nucleic, cytoplasmic and membrane staining
(weak staining = 1, moderate staining = 2 and strong staining = 3)
and the proportion of stained cells (0%-5% = 0, 5%-25% = 1, 26%-
50% = 2, 51%-75% = 3 and 76%-100% = 4). The staining of AMPH1
was remarkably reduced in ovarian cancer tissues compared with
normal ovarian tissues (Figure 6A-C). In addition, the clinicopatho-
logic characteristics of patients and the correlation of AMPH1
demonstrated that AMPH1 was correlated to all the ovarian cancer
patients (Figure 6D). We also detected the staining of p-PI3K and
p-AKT, and the results showed the expression of p-PI3K and p-AKT
increased in ovarian cancer tissues compared with normal ovarian
tissues (Figure 6E). Three types of Skova3 cells (control, sh AMPH1, AMPH-OE) were inoc-
ulated in mice. After 21 days, mice were sacrificed. Tumour volume and
weight in each group were measured. As shown in Figure 5, the absence
of AMPH1 increased tumour volume and tumour weight, whereas over-
expression of AMPH1 decreased tumour volume and tumour weight
(Figure 5A-C). These results suggest that AMPH1 inhibited ovarian
tumour progression in xenograft mouse model. Furthermore, we de-
tected the AMPH1, E-cadherin and PI3K/AKT levels in tumours derived
from cells silenced and high expressed AMPH1 and found the expres-
sion of AMPH1 still decreased in the dissected tumours (Figure 5D). 3.6 | The staining of AMPH1 is decreased in
human ovarian cancer tissues compared with normal
ovarian tissues E, Immunohistochemistry analysis of p-PI3K and p-AKT in human ovarian cancer samples and normal ovarian tissues into ovarian cancer cell lines, Caov-3 and Skov3 cells, to construct
AMPH1 knockdown or AMPH1 overexpression stable cell strains. Our results showed that AMPH1 might function as a tumour sup-
pressor in ovarian cancer via regulating PI3K/AKT signalling pathway. In detail, we demonstrated that AMPH1 inhibited Caov-3 and
Skov3 cells growth. In addition, AMPH1 promoted caspase-3 ac-
tivity, resulting in the increase of cell apoptosis. Ovarian cancer is
one of the most aggressive type's gynaecologic cancers in the world
characterized by the tendency of metastasizing early.13,14 Metastasis
is generally connected with poor prognosis. Thus, we detected the
connection between AMPH1 and cell migration, and further found
that AMH1 prevented cell migration. Xenograft mouse model exper-
iment showed that AMPH1 significantly inhibited ovarian tumour
progression. These findings suggest AMPH1 functions as a tumour
suppressor, which is consistent with previous studies.10,11 However,
different from these studies, we not only utilized AMPH1 knock-
down cell strain, but also enabled AMPH1 overexpression cell strain
to evaluate the association between AMPH1 and ovarian cancer,
which is more comprehensive and convincing. into ovarian cancer cell lines, Caov-3 and Skov3 cells, to construct
AMPH1 knockdown or AMPH1 overexpression stable cell strains. Our results showed that AMPH1 might function as a tumour sup-
pressor in ovarian cancer via regulating PI3K/AKT signalling pathway. the anti-oncogene effects of AMPH1 in ovarian cancer, we evaluated
the association between AMPH1 and p-PI3K or p-AKT. AMPH1 in-
hibited the activation of PI3K/AKT signalling pathway in ovarian can-
cer. 3.6 | The staining of AMPH1 is decreased in
human ovarian cancer tissues compared with normal
ovarian tissues AMPH1, an abundant protein in nerve terminals, plays a critical role
in the recruitment of dynamin to sites of clathrin-mediated endo-
cytosis.3 It is recently reported to be associated with cancer pro-
gression, including breast cancer,10 and lung cancer.11 However,
the impact of AMPH1 on ovarian cancer is unclear. Here, this study
transfected sh AMPH1 or PCMV-AMPH overexpression plasmid To explore AMPH1 expression in patients with ovarian cancer, we
performed immunohistochemical staining for AMPH1, and calcu-
lated the IHC score of AMPH1. Each specimen was scored according CHEN et al. 7658 FI G U R E 6 The staining of AMPH1 was remarkably reduced in human ovarian cancer samples compared with normal ovarian tissues. A, Immunohistochemistry analysis of AMPH1 in human ovarian cancer samples and normal ovarian tissues. B, Each specimen was scored
according to the proportion of stained cells (0%-5% = 0, 5%-25% = 1, 26%-50% = 2, 51%-75% = 3 and 76%-100% = 4). C, Each specimen was
scored according to the intensity of the nucleic, cytoplasmic and membrane staining (weak staining = 1, moderate staining = 2 and strong
staining = 3). IHC score showed that the staining of AMPH1 was remarkably reduced in ovarian cancer tissues compared with normal ovarian
tissues. ***P < .001. D, The clinicopathologic characteristics of patients and the correlation of AMPH1 were shown. P value was determined
by Student's t test. E, Immunohistochemistry analysis of p-PI3K and p-AKT in human ovarian cancer samples and normal ovarian tissues FI G U R E 6 The staining of AMPH1 was remarkably reduced in human ovarian cancer samples compared with normal ovarian tissues. A, Immunohistochemistry analysis of AMPH1 in human ovarian cancer samples and normal ovarian tissues. B, Each specimen was scored
according to the proportion of stained cells (0%-5% = 0, 5%-25% = 1, 26%-50% = 2, 51%-75% = 3 and 76%-100% = 4). C, Each specimen was
scored according to the intensity of the nucleic, cytoplasmic and membrane staining (weak staining = 1, moderate staining = 2 and strong
staining = 3). IHC score showed that the staining of AMPH1 was remarkably reduced in ovarian cancer tissues compared with normal ovarian
tissues. ***P < .001. D, The clinicopathologic characteristics of patients and the correlation of AMPH1 were shown. P value was determined
by Student's t test. ACKNOWLEDGEMENTS This work was supported by from National Natural Science
Foundation of China (81572547). This work was also supported
in part by grants Shanghai Collaborative Innovation Center for
Translational Medicine (TM201923), Shanghai Municipal Key Clinical
Specialty (shslczdzk06302). 11. Yang H, Wan Z, Huang C, Yin H, Song D. AMPH-1 is a tumor sup-
pressor of lung cancer by inhibiting Ras-Raf-MEK-ERK signal path-
way. Lasers Med Sci. 2019;34:473-478. 12. Song Q, Wu JZ, Wang S, Chen ZB. The ABO blood group is an inde-
pendent prognostic factor in patients with ovarian cancer. J Cancer. 2019;10:6754-6760. 13. Binju M, Amaya-Padilla MA, Wan G, Gunosewoyo H, Suryo
Rahmanto Y, Yu Y. Therapeutic inducers of apoptosis in ovarian
cancer. Cancers. 2019;11:1786. AUTHOR CONTRIBUTIONS 15. Jayson GC, Kohn EC, Kitchener HC, Ledermann JA. Ovarian cancer. Lancet. 2014;384:1376-1388. Tianying Zhong and Lihua Wang conceived and designed the experi-
ments; Wenjiao Cao and Yajun Chen performed the experiments
and analysed the data. Yajun Chen, Tianying Zhong and Lihua Wang
wrote the manuscript. Wenjiao Cao and Lihua Wang conducted the
experiments for new figures and provided experimental methods
and data. All authors contributed to data analysis, drafting or revis-
ing the article, gave final approval of the version to be published, and
agree to be accountable for all aspects of the work. 16. Bernard A, Chevrier S, Beltjens F, et al. Cleaved caspase-3 tran-
scriptionally regulates angiogenesis-promoting chemotherapy re-
sistance. Cancer Res. 2019;79:5958-5970. 17. Ogane N, Yasuda M, Kato H, et al. Cleaved caspase-3 expression is
a potential prognostic factor for endometrial cancer with positive
peritoneal cytology. Cytopathology. 2018;29:254-261. 18. Hubbard PA, Moody CL, Murali R. Allosteric modulation of Ras and
the PI3K/AKT/mTOR pathway: emerging therapeutic opportuni-
ties. Front Physiol. 2014;5:478. 19. Gold ES, Morrissette NS, Underhill DM, Guo J, Aderem A. Amphiphysin IIm, a novel amphiphysin II isoform, is required for
macrophage phagocytosis. Immunity. 2000;12:285-292. CONFLICT OF INTEREST The authors declare no competing financial interests. The authors declare no competing financial interests. 14. Zhao G, Cardenas H, Matei D. Ovarian cancer—why lipids matter. Cancers. 2019;11:1870. 6. Bauerfeind R, Takei K, De Camilli P. Amphiphysin I is associated
with coated endocytic intermediates and undergoes stimula-
tion-dependent dephosphorylation in nerve terminals. J Biol Chem. 1997;272:30984-30992. Finally, we used IHC to detect AMPH1 tumours. IHC score re-
sults showed that the staining of AMPH1 was decreased in ovarian
cancer tissues compared with normal ovarian tissues, which is con-
sistent with our results in vitro and in vivo that AMPH1 functions as
a tumour suppressor in ovarian cancer. 7. Dropcho E. Antiamphiphysin antibodies with small-cell lung
carcinoma and paraneoplastic encephalomyelitis. Ann Neurol. 1996;39:659-667. Our study identified AMPH1 as a tumour suppressor in ovarian
cancer. The anti-oncogene effect of AMPH1 may through induce
apoptosis via promoting caspase-3 activity, and suppressing the ac-
tivation of PI3K/AKT signalling pathway. These findings reveal that
AMPH1 may be used as a potential agent for ovarian cancer therapy. 8. Otsuka A, Hirose K, Kilimann MW, Kamata T. Amphiphysin1 inhib-
its vitronectin-mediated cell adhesion, spreading, and migration in
vitro. Biochem Biophys Res Comm. 2003;301:769-775. 9. Jiang L, Ge W, Geng J. miR-425 regulates cell proliferation, migra-
tion and apoptosis by targeting AMPH-1 in non-small-cell lung can-
cer. Pathol Res Pract. 2019;215:152705. 10. Chen Y, Liu J, Li L, Xia H, Lin Z, Zhong T. AMPH-1 is critical for breast
cancer progression. J Cancer. 2018;9:2175-2182. DATA AVAILABILITY STATEMENT All data generated or analysed during this study are included in this
article. 20. Fan S, Bei Z, Ping L, et al. PI3K/AKT/mTOR/p70S6K path-
way is involved in Aβ25-35-induced autophagy. Biomed Res Int. 2015;2015:1-9. 21. Sun R, Zhai R, Ma C, Miao W. Combination of aloin and metformin
enhances the antitumor effect by inhibiting the growth and invasion
and inducing apoptosis and autophagy in hepatocellular carcinoma
through PI3K/AKT/mTOR pathway. Cancer Med. 2020;9:1141-1151. 3.6 | The staining of AMPH1 is decreased in
human ovarian cancer tissues compared with normal
ovarian tissues This pathway is one of many mechanisms that regulate cell cycle
and cell apoptosis, and dysregulation of a component in this pathway
leads to cancer.18 PI3K mainly phosphorylates lipid-based phospha-
tidylinositol secondary messengers upon activation by receptors on
the cell surface, and functions as an important regulator of macro-
phage phagocytosis, and suppression of PI3K inhibits the recruitment
of AMPH to the phagocytic cup.18,19 AKT binds the PIP prodsucts of
PI3K via its pleckstrin homology domain for recruitment to the plasma
membrane.18 In addition, PI3K/AKT signalling pathway is identified
as the primary pathway involved in initiation and regulation of auto-
phagy.20 Autophagy is an intracellular lysosomal pathway, involved
in protein degradation and organelle degradation.21 Interestingly, as
another significant form of programmed cell death, autophagy is fre-
quently deregulated in cancer.22 Autophagy mediates both cell death
promoting and cell death inhibiting activity, which largely depends on
cell types and the magnitude of autophagy.22 However, excessive au-
tophagy causes cell death.23 In this study, AMPH1 inhibited the acti-
vation of PI3K/AKT pathway and might eventually induce tumour cell
death. This is the first time that AMPH1 is reported to regulate PI3K/
AKT signalling pathway. p
g
g
g
g p
y
In detail, we demonstrated that AMPH1 inhibited Caov-3 and
Skov3 cells growth. In addition, AMPH1 promoted caspase-3 ac-
tivity, resulting in the increase of cell apoptosis. Ovarian cancer is
one of the most aggressive type's gynaecologic cancers in the world
characterized by the tendency of metastasizing early.13,14 Metastasis
is generally connected with poor prognosis. Thus, we detected the
connection between AMPH1 and cell migration, and further found
that AMH1 prevented cell migration. Xenograft mouse model exper-
iment showed that AMPH1 significantly inhibited ovarian tumour
progression. These findings suggest AMPH1 functions as a tumour
suppressor, which is consistent with previous studies.10,11 However,
different from these studies, we not only utilized AMPH1 knock-
down cell strain, but also enabled AMPH1 overexpression cell strain
to evaluate the association between AMPH1 and ovarian cancer,
which is more comprehensive and convincing. The PI3K/AKT signalling pathway is one of many mechanisms that
regulate cell cycle and cell apoptosis, and dysregulation of a compo-
nent in this pathway leads to cancer.18 To further investigate whether
PI3K/AKT signalling pathway is involved in the mechanism underlying CHEN et al. 7659 REFERENCES 22. Xue S, Zhou Y, Zhang J, et al. Anemoside B4 exerts anti-cancer
effect by inducing apoptosis and autophagy through inhibiton of
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Does stereopsis account for the link between motor and social skills in adults?
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Danielle Smith
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Spoken
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© The Author(s). 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Smith et al. Molecular Autism (2018) 9:55
https://doi.org/10.1186/s13229-018-0234-4 Smith et al. Molecular Autism (2018) 9:55
https://doi.org/10.1186/s13229-018-0234-4 Open Access Abstract Background: Experimental and longitudinal evidence suggests that motor proficiency plays an important role in
the development of social skills. However, stereopsis, or depth perception, may also play a fundamental role in
social skill development either indirectly through its impact on motor skills or through a more direct route. To date,
no systematic study has investigated the relationship between social skills and motor ability in the general adult
population, and whether poor stereopsis may contribute to this association. This has implications for clinical populations
since research has shown associations between motor abnormalities and social skills, as well as reduced depth perception
in autism spectrum disorder and developmental coordination disorder. Methods: Six hundred fifty adults completed three validated questionnaires, the stereopsis screening inventory, the Adult
Developmental Coordination Disorder Checklist, and the Autism Spectrum Quotient. Results: An exploratory factor analysis on pooled items across all measures revealed 10 factors that were largely
composed of items from a single scale, indicating that any co-occurrence of poor stereopsis, reduced motor proficiency,
and difficulties with social interaction cannot be attributed to a single underlying mechanism. Correlations
between extracted factor scores found associations between motor skill and social skill. Conclusions: Mediation analyses suggested that whilst fine motor skill and coordination explained the
relationship between stereopsis and social skill to some extent, stereopsis nonetheless exerted a substantial direct
effect upon social skill. This is the first study to demonstrate that the functional significance of stereopsis is not
limited to motor ability and may directly impact upon social functioning. Keywords: Stereopsis, Stereoability, Depth perception, Motor skills, Social skills, Factor analysis, Path analysis Does stereopsis account for the link between
motor and social skills in adults? Danielle Smith1, Danielle Ropar2 and Harriet A Allen2* Background behaviours, such as initiation of bids for joint attention and
directed gestures [6, 7]. Other research, using longitudinal
designs, reported relationships between motor function at
5–6 years and a range of social behaviours at 6–7 years [8],
and between motor abilities at 6–7 years and social status
with peers at 9–10 years [9]. Additionally, a reduction in so-
cial play and increased social reticence has been noted in
children with poor motor skills [10]. Motor ability exhibits rich and complex relationships with
regards to other cognitive domains [1]. A basic movement
repertoire of functional actions involving both fine (such
as pointing a finger, eye movements) and gross (such as
arm gestures, walking together) motor domains aids in
the initiation and sustainment of successful social interac-
tions [2]. For instance, motor control plays an important
role in joint attention (e.g. head-turning, reaching, point-
ing) and imitation [3], both crucial components of social
relations [4]. A relationship between social and motor abil-
ities has been identified in typically developing children as
young as 8 months [5], with development from crawling to
walking encouraging the use of more advanced social Although there appears to be sufficient evidence to
support a link between motor and social skills, our un-
derstanding of this relationship still has important gaps
that need to be addressed. Firstly, although evidence for
a relationship between motor and social skills has been
demonstrated in several studies with typically developing
children, it remains unclear whether this relationship
would extend into early adulthood. Secondly, previous
literature exploring the link between motor and social * Correspondence: h.a.allen@nottingham.ac.uk
2School of Psychology, University of Nottingham, Nottingham NG7 2RD, UK
Full list of author information is available at the end of the article * Correspondence: h.a.allen@nottingham.ac.uk
2School of Psychology, University of Nottingham, Nottingham NG7 2RD, UK
Full list of author information is available at the end of the article Page 2 of 15 Page 2 of 15 Smith et al. Molecular Autism (2018) 9:55 Smith et al. Molecular Autism (2018) 9:55 Smith et al. Molecular Autism functioning has neglected important visual skills, such as
depth perception, which may help contribute to the rela-
tionship between these two domains. For example, there
are clear links between depth perception in terms of
stereoacuity (the estimation of depth from combining in-
formation from two eyes) and the physical manipulation
of objects, which may help explain poor coordination or
clumsiness. More specifically, the shape of the hand is
wider and less accurate when reaching and grasping, and
the time taken for the reach is much slower in individ-
uals where stereopsis is reduced or absent in adults [11–
14], with even larger errors in these tasks for children
with reduced stereoacuity [15–17]. Poor depth percep-
tion can also impact upon gross motor skills such as
walking; adults with reduced stereoacuity also demon-
strate differences in gait with a more cautious approach,
higher toe clearance, and increased hesitation [18, 19]. All of these skills may have implications for the likeli-
hood of taking part in team, motor, and social activities;
however, this relationship has rarely been studied. in recent years. Both clinical and screening studies have
reported significant relationships between motor abilities
and parent-reported peer or social problems [38–41],
showing children with impaired motor skills engaging in
more solitary-type activities and generally being more iso-
lated from their peers. There is also evidence from paren-
tal report and empirical work that individuals with autism
show violations of personal space with others which might
impact upon social acceptance [42]. Finding a relationship
between stereopsis, motor ability, and social ability might
provide a useful avenue for understanding, and perhaps
treating these developmental disorders. g
p
From the few studies that have made a direct compari-
son between ASD and DCD, it would appear that both
disorders exhibit a similar range of social and motor diffi-
culties [43, 44]. Importantly, however, it may be that the
co-occurrence of these impairments is attributable to an-
other underlying factor. The majority of studies reporting
stereoacuity in ASD indicate that those with ASD are less
sensitive to binocular disparity than their TD counterparts
or normative data [45–51]. There is one contradictory
finding. Milne, Griffiths, Buckley, and Scope [52] used the
Frisby stereotest and found no significant group difference
in stereoacuity between the TD and ASD groups. How-
ever, Anketell et al. [46, 47] found differences using this
stereotest. A general stereopsis deficit has also been ob-
served in those with DCD; Creavin, Lingam, Northstone,
and Williams [53] reported that those with DCD were on
average 8 percentage points more likely to have impaired
stereopsis (i.e. stereoacuity of higher than 60 arc s) than
their TD peers (a 44.5% relative increase), and those with
severe DCD were more likely to show evidence of poor
depth perception than those with moderate DCD. Depth perception and stereopsis may also impact on so-
cial skills more directly by influencing social behaviour,
perhaps via social norms. When interacting with another
individual, we need to determine and maintain an appro-
priate amount of personal space. Stereo depth cues are
most useful in this peri-personal space [20] suggesting
those with reduced ability to judge this might inadvert-
ently violate these norms. Good stereo ability requires
good alignment and vergence of the eyes, but there is also
evidence that those with strabismus (i.e. poor alignment
of the eyes) experience social exclusion [21]. There are
also preliminary links between poorly regulated eye con-
tact and social abilities [22]. Finally, Kuang et al. [23]
found a relationship between stereopsis and quality of life
in older people, but the mechanisms for this are, as yet,
unclear. There are, therefore several ways in which depth
perception might impact upon social skills either indir-
ectly through motor skills, or through a more direct route. To date, however, research has only investigated the links
between motor and social ability and the links between
motor and visual abilities separately. Stereoacuity The Stereopsis Screening Inventory (SSI) is a self-report
screening inventory for stereopsis [56]. It is composed of
10 statements with 5 response options (never, seldom, oc-
casionally, frequently, and always). Coren and Hakstian
[56] demonstrated that the scores obtained using the SSI
correlate highly (r = .8) with laboratory measures of stere-
opsis such as the TNO test, with others demonstrating a
moderate relationship [59] between these measures
(r = .34). Recommended cut-offs are 17 for moderate
stereopsis deficit and 30 for major stereopsis deficit. Potential participants were provided with a paragraph
explaining the study and a hyper-link taking them to the
survey website. Although all materials used were originally
developed as ‘pen-and-paper’ questionnaires, it appears
there is little variation in responses when questionnaires
are presented on-line [54, 55]. Individuals were advised
the completion of the study would take approximately
20 min. All participants were offered the chance to enter
into a prize draw for one of two £15 vouchers. The Autostereogram Self-Assessment (ASA) is a short
four-item survey created for the present study where the
participant self-assesses their autostereogram skill. Based
upon short reports by Wilmer and Backus [59] and
Cisarik, Davis, Kindy, and Butterfield [60], the questions
asked the subject to identify two autostereograms (Fig. 1). The respondent was offered four possible choices plus an
‘I don’t know’ option. Correct answers were designated a
score of 1, all other answers were given a 0. Respondents
were also asked how difficult they found viewing the auto-
stereograms (on a scale from 1 to 5, where 1 was ex-
tremely difficult and 5 very easy), and whether they had
successfully perceived stereopsis in an autostereogram
previously (‘yes’ answers were given a score of 1, all others
a 0). Self-reported skill to perceive depth in autostereo-
grams has been found to be predictive of stereoacuity, as
measured by the TNO test (r = .45; [59, 60]). The sample included 650 participants aged between 16
and 70 (mean 26.46 ± 10) years. Demographic data in
the form of gender, age, and occupation were collected,
though these were optional. There were 227 males (age
27.01 ± 10.31; range = 16–67 years), 369 females (age
26.24 ± 9.79; range = 16–70 years), and 6 who identified
as “other” (age 25.33 ± 4.23; range = 21–33 years). The current study
h Despite the suggestion from within clinical groups that
poor depth perception, or stereopsis, may be linked to
both motor skills and social abilities, there has been little
research to investigate the relationship between these
three abilities. Furthermore, previous work that has in-
vestigated the relationship between motor and social
skills has focused on children, with a remarkable paucity
of research involving adults. It is essential to initially es-
tablish the links between stereopsis, motor, and social
skills in a larger general population where these skills
vary before exploring these relationships in clinical pop-
ulations, such as ASD, which are often complicated with
other co-occurring conditions. Through measuring aut-
istic traits in a large sample within the general popula-
tion, this study will be able to identify any potential
links, either direct or indirect, between depth perception
and social skills. This work will help define more specific
questions to explore this area further in ASD popula-
tions. This study has two primary aims. The first is to The potential relationship between motor, social, and
visual abilities has important implications for understand-
ing developmental disorders [24–26]. Poor social skills are
central to the diagnostic criteria of ASD [27]. Gross and
fine motor impairment as well as difficulties in motor
planning have been reported in up to 90% of those with
ASD [28–33]. Significant correlations between motor
skills and socialisation [34] and degree of social impair-
ment [35–37] have been found in children with ASD. Motor dysfunction is also central to the diagnosis of DCD
(also referred to as ‘dyspraxia’ and affects around 5% of
the population [31]), and there has been an increasing
interest in the social functioning of individuals with DCD Page 3 of 15 Smith et al. Molecular Autism (2018) 9:55 Smith et al. Molecular Autism (2018) 9:55 Smith et al. Molecular Autism diagnosed with an ASD (all self-described as Asperger’s
syndrome), and 1.85% (n = 12) reported amblyopia,
strabismus or general “poor vision”. All diagnostic dis-
closures are summarised in Additional file 2: Table S2. extend the previous research linking motor and social
skill impairment in children to a typical adult sample,
identifying the possible later consequences of early deficit
in these domains. Materials The questionnaires were presented in a fixed order, begin-
ning with the Stereopsis Screening Inventory [56], then the
autostereogram self-asssessment, followed by the Adult De-
velopmental Disorder Checklist [57] and the Autism
Spectrum Quotient [58]. Finally, demographics including
age and gender were requested, but these were optional. Participants and recruitment Ethical permission from the University of Nottingham’s
School of Psychology Ethics Committee was granted prior
to recruitment. Participants were sampled opportunistic-
ally from Reddit (www.reddit.com; n = 311, 47.8%), social
media and email (n = 193, 29.7%), and an internal recruit-
ment system for undergraduate students at the University
of Nottingham for partial completion of course credit
(n = 146, 22.5%). The current study
h Secondly, to examine which particular
aspects of motor and social skill impairment were contrib-
uted to by reduced stereopsis; that is, if the effects of poor
stereoacuity are strong enough to be able to affect social
skill either directly or through mediation by motor ability. Stereoacuity The
age values were not of a normal univariate distribution
for any gender group; all groups had positively skewed
age distributions, and the ages of the “male” and “other”
genders were platykurtic. However, age distributions for
each group were roughly equivalent according to a
two-sample Kolmogorov-Smirnov test (male vs female
D = 0.1, p = 0.08; male vs other D = 0.3, p = 0.6; female vs
other D = 0.4, p = 0.2). Motor skills Motor skills
The
Adult
Developmental
Coordination
Disorder
Checklist (ADC) is a validated screening tool for identi-
fying the difficulties experienced by adults with DCD
[57]. The ADC consists of three sub-scales; the first re-
lates to difficulties that the individual experienced as a
child (10 items). The second (10 items) and third
sub-scales (20 items) relate to current difficulties. The
second sub-scale focuses on the individual’s perception
of their performance, whereas the third sub-scale relates
to current feelings about their performance as reflected
upon by others. All items are rated on a four-point scale Of
the
participants
who
reported
an
occupation
(89.5%, n = 582), 54.1% (n = 315) reported that they were
enrolled in secondary or tertiary education, 38% (n = 221)
were in employment, and 6.19% (n = 36) were not in
work or education. More details regarding occupation,
including a breakdown by industry, can be seen in
Additional file 1: Table S1. Diagnoses were not col-
lected as part of the demographical data. However, a
number of participants (3.69%, n = 24) disclosed various
psychiatric or organic illness via the feedback section of
the questionnaire. 0.92% (n = 6) reported they had been Page 4 of 15 Smith et al. Molecular Autism (2018) 9:55 Smith et al. Molecular Autism Fig. 1 The two autostereograms used in the current study. The top
autostereogram contains a shark [115], and the bottom a teapot
[116]. The instructions for viewing are as follows: “Above is an
autostereogram or Magic Eye© picture - to reveal the hidden 3D
illusion, you must diverge your eyes (i.e. focus beyond the image). First, bring your face close to the page (so that you are almost touching
it with your nose). The image should appear blurry. Focus as though you
are looking through the image into the distance. Very slowly move away
from the page until you begin to perceive depth in the image. At this
point, hold very still and the hidden image will slowly appear” of AQ items have been inconsistent, with studies finding
two, three or four factors rather than five [63]. of AQ items have been inconsistent, with studies finding
two, three or four factors rather than five [63]. Data analysis
l Statistical analyses were performed using R 3.0.1. The
relationship between scores on the ADC, AQ, ASA, and
SSI were first examined using Pearson correlation ana-
lysis. The data were then randomly split into two equally
sized groups (n = 325) to act as training and test data in
a cross validation procedure. All items from all measures
(minus the two ADC items mentioned above) within the
training data set were subjected to exploratory factor
analysis (EFA). Oblique rotation was specified for the
EFA, given that the factors were expected to correlate
with one another based on theoretical and empirical
grounds [67]. Parallel analysis and Velicier’s Minimum
Average Partial Test (available as part of the psych pack-
age) were used to determine the number of factors to re-
tain. Factors were further interpreted if the grouping of
the loading variables made conceptual sense. Given the
fairly large sample size, items were considered to load
onto a factor if their loading was ≥.32 [68]. Missing data If a participant left more than 10% of responses across
all items blank, the data were excluded from the analysis
(n = 0; highest proportion of missing data for a single par-
ticipant was 8.308%). The proportion of missing data for
any individual questionnaire item ranged from 0 to 30%. Closer inspection of the pattern of missingness revealed
that two items relating to driving ability in the Adult
Developmental Coordination Disorder Checklist (“Did it
take you longer than others to learn to drive?” and “If you
are a driver, do you have difficulty parking a car?”)
accounted for the highest amount of missing data
(28.154% and 30% respectively). When the data from these
questions were removed from the analysis, the highest
proportion of missing data for a single item was reduced
to 10.154%. Although the number of subjects in the study was 650,
290 cases were missing a response for at least one item. Homoscedasticy of the data was tested using the TestM-
CARNormality function [64], which is part of the Mis-
sMech package in R. The test of homoscedasticy was
rejected, indicating that the data was not missing com-
pletely at random (MCAR). The R package missForest
[65] was used to impute the missing data. This has been
demonstrated to introduce the least imputation error
and has the smallest prediction difference from actual
non-imputed values [66]. Fig. 1 The two autostereograms used in the current study. The top
autostereogram contains a shark [115], and the bottom a teapot
[116]. The instructions for viewing are as follows: “Above is an
autostereogram or Magic Eye© picture - to reveal the hidden 3D
illusion, you must diverge your eyes (i.e. focus beyond the image). First, bring your face close to the page (so that you are almost touching
it with your nose). The image should appear blurry. Focus as though you
are looking through the image into the distance. Very slowly move away
from the page until you begin to perceive depth in the image. At this
point, hold very still and the hidden image will slowly appear” (never, sometimes, frequently or always), resulting in
possible scores ranging from 0 to 120. Recommended
cut-off scores include 56 for “at risk of DCD” and 65 for
“probable DCD” [57]. The latent structure of the ADC
has not yet been confirmed using factor analysis. Social or autism-related traits The Autism Spectrum Quotient (AQ) is a self-report
questionnaire comprising of 50 statements [58]. It was
designed as a measure of autistic characteristics in the
general population. Although a 4-point response format
is used, it is typically scored in a binary manner, where a
response is scored as a one if it indicates an autistic trait
and zero if this is not the case; this yields a score that
can range from 0 to 50. Using this scoring approach,
Baron-Cohen et al. [58] determined the optimal cut-off
for identifying people with clinically significant levels of
autistic traits to be 32 or above. The AQ can also be
scored according to the 4-point response option [61, 62],
which potentially yields a more sensitive index of ASD
severity. In the current study, binary scoring was used to
determine the proportion of participants that scored
above the 32-point threshold mentioned previously. For
all other analyses, including the exploratory factor ana-
lysis, the 4-point response was used. Past factor analyses Cross-validation was then performed using the test
data set, with the factors extracted using EFA being used
to specify the factor structure for confirmatory factor
analysis (CFA). In CFA, there is no single definitive Page 5 of 15 Smith et al. Molecular Autism (2018) 9:55 Smith et al. Molecular Autism [76, 77] and stereopsis deficit 40% [78]); however, one
must be cautious when comparing rates of diagnosis in
the clinic to questionnaire based estimates, see the
“Discussion” section. It was not uncommon for partici-
pants who had a score above threshold for one measure to
also score above threshold for at least one of the other
measures (see Additional files 1, 2, and 3). indicator of model fit. The overall model fit was there-
fore assessed in terms of five measures from two per-
spectives: absolute fit and comparative fit to a base
model, with index cut-offs (seen in brackets) informed
by recommendations in the literature [69–72]. Absolute
fit measures included the model chi-square/degrees of
freedom (χ2/df; 3.0), standardised root mean square re-
sidual (SRMR; .08), and root mean square error of ap-
proximation (RMSEA; .06). The comparative measures
were comparative fit index (CFI; .9) and Tucker-Lewis
index (TLI; .9). Post hoc modification indices were ap-
plied to improve model fit. Social or autism-related traits These indices were only used
when modifications could be supported with theory as
suggested by the literature; here, modifications consisted
of allowing correlated residuals between items that
loaded on to the same factor [73]. Descriptive statistics Tests of multi- and uni-variate normality indicated that the
scores across all items did not meet the assumption of nor-
mality (Royston’s H test [74]; H = 11,580.473, p = < 0.001). For large sample sizes, significant results can be derived
even in the case of a small deviation from normality [75]. Correlation of measure totals Bivariate correlations of measure scores revealed a num-
ber of significant associations. A strong positive relation-
ship was observed between AQ and ADC total scores
(r(648) = 0.628, p = < 0.001), meaning that those with
higher levels of autistic traits were also likely to exhibit
higher
levels
of
dyspraxic
traits. Small-to-moderate
positive correlations were observed between SSI, and both
AQ (r(648) = 0.277, p = < 0.001) and ADC (r(648) = 0.268,
p = < 0.001) scores, indicating that higher levels of autistic
and dyspraxic traits were associated with an increased de-
gree of stereoscopic deficit. A small negative relationship
was also observed between ASA and ADC scores (r(648)
= −0.106, p = 0.007), denoting that those with increased
dyspraxic traits tended to be worse at perceiving autoster-
eograms. No significant relationship was found between
ASA and either AQ (p = 0.056) or (surprisingly) SSI scores
(p = 0.502). In the case where CFA fit indices indicated an ad-
equate fit to the test data, bivariate correlations and sub-
sequent moderation and mediation analyses in the form
of structural equation modelling were conducted upon
the extracted factor scores from the CFA to determine
how they related to one another. Exploratory factor analysis The aim of this study was to assess the existence of la-
tent variables, thus EFA was used to determine the di-
mensional structure of pooled items across the four
measures previously described. All scales demonstrated acceptable internal consistency—
see Table 1 for these and other descriptive data including
the percentage of the total sample who met cut-off scores
indicating clinically significant impairment for each meas-
ure. Of note is a higher incidence than would be expected
of participants meeting cut-offs for clinically significant
impairment for each standardised measure. These are
higher incidences than would be expected from partici-
pants drawn from the general population (where DCD has
a prevalence of approximately 5% [31]), ASD 1.1–2.4% For the training dataset (n = 325) the Kaiser-Meyer-
Olkin coefficient of sampling adequacy was good (.857; .6
is recommended by Cerny and Kaiser [79]) and Bartlett’s
test of
sphericity [80]
was significant (χ2
(5151) =
17,439.845, p = < 0.001), indicating that the data were suit-
able for factor analysis. Parallel analysis [81] and Velicer’s
minimum average partial test [82] recommended that 10
factors be extracted from the data [83, 84]. Factor loadings Table 1 Descriptive statistics for the Adult Developmental Coordination Disorder Checklist (ADC), Autism Spectrum Quotient (AQ),
autostereogram self-assessment (ASA), and Stereopsis Screening Inventory (SSI) (n = 650). Clinically significant impairment is based
on Coren and Hakstian [56] Kirby et al [57] and Baron-Cohen e al [58] on Coren and Hakstian [56], Kirby et al. [57], and Baron Cohen e al. [58]
M (SD)
Range
Cut-off scores indicating
clinically significant impairment
% of participants meeting
cut-off
Skewness
Kurtosis
Cronbach’s α
ADC
41.7 (21.97)
0–116
“DCD at risk” = 56–64
2.31%
0.75
3.09
0.94
“Probable DCD” = ≥65
13.08%
AQ
121.52 (22.5)
78–179
≥32
24.92%; note, 6 participants
disclosed ASD diagnosis
0.29
2.16
0.91
ASA
3.6 (2.53)
1–8
Data not available
N/A
0.61
1.82
0.72
SSI
24.14 (9.28)
9–45
Moderate deficit = 17–29
34.62%
−0.07
1.78
0.87
Major deficit = ≥30
35.08% Page 6 of 15 Smith et al. Molecular Autism (2018) 9:55 Smith et al. Molecular Autism isolation due to motor proficiency, and social skill factor
scores were retained. As can be seen in Table 3, the major-
ity of these factors showed medium-to-large correlations
with one another. All mediation analyses reported here
were performed using lavaan’s structural equation model-
ling (SEM) framework. Confirmatory factor analysis The factor structure suggested by EFA was cross-validated
by means of CFA, using the lavaan package. The ‘test’ data
(n = 325) were analysed using the MLR estimator, which is
robust to the non-normality of the observed variables
[85]. In the first model, items (indicators in CFA termin-
ology) which had a sufficiently high factor loading in the
initial EFA (≥.32) were estimated as free parameters; all
other items were fixed to zero. The factors (or latent vari-
ables) were allowed to covary freely. Though the initial
model showed a reasonable fit on some of the indicators,
it did not meet criteria for acceptable fit for the compara-
tive fit indices (χ2/df = 2.013, CFI = 0.782, TLI = 0.771,
SRMR = 0.08, RMSEA = 0.056). This is to be expected, as
the initial model to be tested through CFA had more
stringent restrictions than the factor model obtained
through EFA, where no factor loadings were fixed to zero. In studies using cross-validation procedures such as those
performed here, it is recommended that a less constrained
model is tested where some parameters are freed [86]. Modification indices were allowed in the creation of an
adjusted model, though with restrictions upon which
changes could be reasonably made to the initial model. Motor skills mediate the link between stereopsis and
isolation To investigate why individuals with worse stereopsis re-
ported increased isolation due to motor proficiency, a
multiple mediation analysis was performed with the me-
diator variables being fine motor skill and coordination. A significant total effect of stereopsis on isolation
emerged, β = 0.179, z = 3.47, p = < 0.001. When dividing
this total effect into the direct effect of stereopsis, and
the total indirect effects of both mediators, the direct
effect of stereopsis was no longer significant, β = 0.02,
z = 0.772, p = 0.44, but the total indirect effect was signifi-
cant, β = 0.158, z = 3.545, p = < 0.001. Both mediator vari-
ables contributed significantly to the indirect effect of
stereopsis upon isolation due to motor proficiency, though
coordination exhibited a greater proportion of mediation
(75.3% of the total effect; β = 0.134, z = 3.471, p = < 0.001)
than fine motor skills (13.273% of the total effect;
β = 0.024, z = 2.294, p = 0.02). After modification indices were applied, where the resid-
uals between indicators loading on to the same latent vari-
able were allowed to correlate with one another if this
significantly improved the fit of the model, all indices indi-
cated an acceptable fit (χ2/df = 1.485, CFI = 0.899, TLI = 0.89,
SRMR = 0.069, RMSEA = 0.039). A scaled chi-square
difference test [87] showed that this modification-
index-adjusted model exhibited a significantly better fit
compared to the initial model (Δχ2(88) = 1261.05,
p = < 0.001). Factor scores were calculated from the ad-
justed CFA model using simple regression [88] for each
participant. These scores were then used to perform medi-
ation analyses in order to better understand the relation-
ships between the factors or latent variables. Exploratory factor analysis were calculated using principal axis factoring with oblmin
(oblique) rotation on 102 of 104 Likert scale questions
across all four measures (omitting the two items of the
ADC which had a high proportion of missing data, see the
“Methods” section) and are shown in Table 2. Labels have
been provided for the 10 extracted factors, based on an in-
terpretation of the items that constitute them; ‘social skill’,
‘stereopsis’, ‘attention to detail’, ‘fine motor skill’, ‘organisa-
tion’, ‘Magic Eye proficiency’, ‘isolation due to motor profi-
ciency’, ‘coordination’, ‘imagination’, and ‘multitasking’. All
items loading on to these factors are shown in Table 2. Motor skills may mediate the link between stereopsis and
social skills Fine motor skill, coordination, and isolation due to
motor proficiency were entered into a multiple medi-
ation analysis to investigate the relationship between
stereopsis impairment and reduced social ability. A sig-
nificant total effect of stereopsis on social skills emerged,
β = −0.312, z = −6.143, p = < 0.001. When dividing this
total effect into the direct effect of stereopsis, and the
total indirect effects of all three mediators, the direct ef-
fect of stereopsis remained significant after adjusting for
all three mediators, β = −0.216, z = −4.615, p = < 0.001. The total indirect effect was also significant, β = −0.096,
z = −3.441, p = < 0.001. Of the three mediator variables,
only fine motor skill contributed significantly to the in-
direct effect of stereopsis upon social skills (12.436% of
the total effect; β = −0.039, z = −2.276, p = 0.02). Neither
coordination nor isolation exhibited a significant amount
of mediation (p = 0.061 and 0.178, respectively). Isolation may mediate the link between coordination/fine
motor skills and social skills Two final mediation models indicated that isolation due to
motor proficiency was a significant mediator both in the re-
lationship between coordination and social skills (39.772%
of the total effect; β = −0.211, z = −2.533, p = 0.01) and fine
motor skill and social skills (40.941% of the total Mediation Factors were only included in this aspect of the analysis
where strong a-priori hypotheses could be made: stereop-
sis, Magic Eye proficiency, fine motor skill, coordination, Page 7 of 15 Page 7 of 15 Smith et al. Molecular Autism (2018) 9:55 Table 2 Factor loadings of a 10-factor EFA solution for items pooled across all measures. Principal axis factoring, oblmin rotation. Loadings below .32 (which explain less than 10% of the variance in that item) are not highlighted and are considered to be
negligible loadings for the purposes of analysis Table 2 Factor loadings of a 10-factor EFA solution for items pooled across all measures. Principal axis factoring, oblmin rotation. Mediation Molecular Autism (2018) 9:55 Table 2 Factor loadings of a 10-factor EFA solution for items pooled across all measures. Principal axis factoring, oblmin rotation. Loadings below .32 (which explain less than 10% of the variance in that item) are not highlighted and are considered to be
negligible loadings for the purposes of analysis (Continued) Table 2 Factor loadings of a 10-factor EFA solution for items pooled across all measures. Principal axis factoring, oblmin rotation. Mediation Loadings below .32 (which explain less than 10% of the variance in that item) are not highlighted and are considered to be
negligible loadings for the purposes of analysis negligible loadings for the purposes of analysis
Measure Item
Social
Stereo Detail Fine motor Org
Magic Eye Isolation Coord Imagine Multi
AQ
Enjoy social chitchat
0.70
0.01
−0.06
−0.09
−0.06 −0.03
−0.04
0.07
0.01
0.08
AQ
Good at social chitchat
0.70
0.04
0.02
−0.05
0.03
0.12
0.03
0.01
−0.05
−0.03
AQ
Find social situations easy
0.67
−0.05
0.07
−0.01
0.01
0.03
0.01
0.00
0.02
−0.08
AQ
Prefer people over things
0.59
−0.01
−0.12
−0.04
0.02
0.04
−0.01
−0.04
0.04
0.09
AQ
Enjoy social occasions
0.56
−0.07
−0.10
−0.14
0.02
−0.02
−0.10
−0.09
0.08
0.01
AQ
Enjoy meeting new people
0.53
−0.07
−0.12
0.02
0.00
0.02
−0.04
−0.20
0.05
0.04
ADC
Choose to spend leisure time on own
−0.48 0.05
0.09
0.20
−0.02 0.02
0.25
−0.01
0.01
0.06
AQ
Easily keep track of several conversations
0.44
−0.03
0.15
0.00
−0.08 −0.01
0.01
0.07
0.19
−0.19
AQ
Can work out what someone is feeling
from their face
0.43
−0.02
0.00
−0.06
−0.08 −0.01
−0.02
0.06
0.34
−0.10
AQ
Prefer to do things with others
0.43
0.05
0.02
0.13
−0.11 −0.02
−0.11
0.02
−0.22
−0.05
AQ
Find it hard to make new friends
−0.41 −0.06
0.30
−0.05
−0.04 −0.06
0.20
0.15
−0.16
−0.06
AQ
New situations bring on anxiety
−0.35 0.08
0.11
−0.11
0.01
0.06
0.17
0.09
−0.08
0.24
AQ
Don’t know how to keep conversation going
−0.33 −0.04
0.22
0.04
0.02
−0.09
0.11
−0.00
−0.25
0.11
AQ
Can easily ‘read between the lines’
0.32
−0.03
0.02
−0.10
−0.03 0.10
0.06
0.03
0.31
−0.14
SSI
Do you think you need glasses
0.03
0.94
0.02
0.05
0.01
−0.01
−0.03
−0.03
0.03
0.01
SSI
Glasses/contact lens wearer
−0.05
0.90
−0.00
−0.00
0.01
0.02
−0.02
−0.02
0.02
−0.00
SSI
W/out correction, clearness of vision in LEFT eye
0.08
0.89
0.02
0.03
−0.04 −0.01
0.07
−0.05
−0.04
0.05
SSI
W/out correction, clearness of vision in RIGHT eye −0.08
0.88
−0.06
−0.03
0.00
0.02
−0.02
0.03
0.06
−0.03
SSI
Vision as good as other people’s
0.06
0.87
0.02
−0.03
0.03
−0.04
−0.00
−0.00
−0.06
−0.01
SSI
Correction needed for reading
−0.02
0.53
0.01
−0.04
−0.02 0.05
−0.09
0.21
−0.05
−0.08
AQ
Notice patterns in things all the time
−0.13
0.05
0.67
−0.08
0.10
0.02
−0.03
−0.08
0.05
−0.05
AQ
Notice car number plates or similar
−0.01
0.02
0.56
−0.03
0.04
0.06
0.03
−0.01
−0.02
−0.07
AQ
Tend to notice details that others do not
−0.14
−0.02
0.55
0.12
0.06
−0.00
−0.10
0.01
0.30
−0.09
AQ
Strong interests, get upset if can’t pursue
0.01
0.04
0.55
0.14
−0.02 −0.04
0.05
−0.03
0.03
0.16
AQ
Notice small sounds
−0.13
0.07
0.47
0.07
0.07
0.06
0.01
0.05
0.18
0.07
AQ
Get strongly absorbed in one thing
−0.24
0.12
0.46
−0.06
0.18
−0.02
−0.02
−0.01
−0.03
−0.02
AQ
Enjoy collecting information about categories
−0.01
−0.07
0.45
0.13
−0.08 0.02
0.04
0.09
−0.07
0.05
AQ
Repetitive topic of conversation
0.06
0.03
0.45
0.11
0.03
−0.01
0.13
−0.01
−0.18
0.01
AQ
Tend to dominate conversation
0.18
0.03
0.43
0.05
−0.06 0.06
0.09
−0.02
−0.04
0.12
AQ
Fascinated by numbers
−0.07
−0.05
0.41
0.07
0.03
0.03
−0.07
0.04
−0.08
−0.06
AQ
Difficult to work out people’s intentions
−0.06
0.06
0.39
−0.00
0.04
−0.03
0.06
−0.00
−0.29
0.16
AQ
Difficulty imagining being someone else
0.08
−0.09
0.37
0.14
−0.01 −0.12
0.09
−0.06
−0.16
0.19
AQ
Say impolite things without realising
0.09
−0.02
0.36
0.14
−0.12 −0.03
0.12
0.02
−0.04
0.25
AQ
Difficulty speaking in turns on phone
−0.01
0.03
0.35
0.13
0.07
−0.05
0.10
0.09
−0.05
0.10
AQ
Difficultly working out characters’
intentions in story
0.12
0.01
0.34
0.06
−0.04 0.14
0.13
0.13
−0.24
0.02
ADC
Others find it difficult to read your writing
0.01
0.04
0.00
0.79
−0.07 0.01
0.05
−0.07
0.01
−0.06
ADC
Difficulty with writing neatly AND quickly
−0.06
0.02
−0.03
0.73
0.03
−0.04
0.02
−0.02
0.03
0.09
ADC
Difficulty with neat writing when child
−0.07
−0.01
0.07
0.70
0.13
−0.04
0.07
−0.11
0.00
0.00
ADC
Difficulties reading own writing
−0.04
0.07
−0.05
0.66
−0.04 0.03
−0.08
0.18
−0.01
−0.15
ADC
Difficulties with writing as fast as peers
0.00
−0.04
0.07
0.65
0.06
0.03
−0.04
0.09
−0.12
0.05
ADC
Difficulty with fast writing as child
0.04
−0.06
0.06
0.62
0.15
0.03
0.01
0.06
−0.07
0.06 Page 8 of 15 Page 8 of 15 Smith et al. Mediation Molecular Autism (2018) 9:55 Table 2 Factor loadings of a 10-factor EFA solution for items pooled across all measures. Principal axis factoring, oblmin rotation. Loadings below .32 (which explain less than 10% of the variance in that item) are not highlighted and are considered to be
negligible loadings for the purposes of analysis (Continued) Table 2 Factor loadings of a 10-factor EFA solution for items pooled across all measures. Principal axis factoring, oblmin rotation. Path analysis The above mediation models were aggregated into a lar-
ger path model. This final model included relationships
with Magic Eye proficiency as detailed in Table 3. This
model had a good fit, with χ2/df = 0.418, CFI = 1, TLI =
1.011, SRMR = 0.017, and RMSEA = < 0.001. The results
of
the
path
analysis
with
standardised
regression Mediation 2. The relationship be-
tween stereopsis and social skills, as well as stereopsis
and isolation (both mediated by fine motor skill and co-
ordination) held in this larger model. The effect of isola-
tion due to motor proficiency acting as a mediator
between fine motor skill/coordination and social skills
did not hold in this larger model. Whilst fine motor skill
and coordination were responsible for full mediation of
the relationship between stereopsis and isolation due to
motor proficiency, there was no serial mediation from
the fine motor/coordination variables to social skills via
the isolation variable. Finally, Magic Eye proficiency was
not a significant independent variable within the context
of the path model. effect; β = −0.214, z = −4.477, p = < 0.001). Partial me-
diation occurred in both cases, as coordination and
fine motor skill were still significant predictors of so-
cial skills after adjusting for the indirect effect of iso-
lation (coordination: β = −0.32, z = −3.647, p = < 0.001,
fine motor skills: β = −0.309, z = −4.605, p = < 0.001). coefficients are presented in Fig. 2. The relationship be-
tween stereopsis and social skills, as well as stereopsis
and isolation (both mediated by fine motor skill and co-
ordination) held in this larger model. The effect of isola-
tion due to motor proficiency acting as a mediator
between fine motor skill/coordination and social skills
did not hold in this larger model. Whilst fine motor skill
and coordination were responsible for full mediation of
the relationship between stereopsis and isolation due to
motor proficiency, there was no serial mediation from
the fine motor/coordination variables to social skills via
the isolation variable. Finally, Magic Eye proficiency was
not a significant independent variable within the context
of the path model. Mediation Loadings below .32 (which explain less than 10% of the variance in that item) are not highlighted and are considered to be
negligible loadings for the purposes of analysis (Continued) negligible loadings for the purposes of analysis (Continued)
Measure Item
Social
Stereo Detail Fine motor Org
Magic Eye Isolation Coord Imagine Multi
ADC
Difficulty copying without mistakes
−0.09
−0.06
−0.09
0.43
0.10
−0.02
−0.15
0.27
0.04
0.08
ADC
Difficulty with organisation
−0.04
0.07
−0.06
0.14
0.71
0.07
−0.09
0.02
−0.00
−0.03
ADC
Difficulties with organisation as child
−0.01
0.09
−0.05
0.06
0.68
−0.02
0.10
−0.06
−0.01
−0.09
ADC
Others call you disorganised
0.06
0.00
0.11
0.08
0.66
0.09
0.01
0.02
−0.04
0.00
ADC
Tend to lose possessions
0.01
−0.09
−0.03
0.04
0.58
−0.04
0.15
0.03
0.05
0.03
ADC
Difficulty sitting still
−0.08
−0.07
0.23
0.05
0.52
−0.04
−0.02
−0.05
0.07
0.18
ADC
Difficulty planning ahead
−0.08
0.03
0.09
−0.02
0.48
0.04
−0.14
0.02
−0.18
0.29
ADC
Bump into, spill, or break things
−0.00
−0.07
−0.02
−0.04
0.43
−0.10
0.41
0.19
0.04
0.02
ADC
Difficulty managing money
−0.02
−0.04
0.05
0.01
0.43
0.04
−0.17
0.23
−0.03
0.15
ADC
Can lose attention in certain situations
−0.02
0.05
0.12
0.02
0.42
−0.02
−0.05
0.04
−0.07
0.31
ADC
Bumped into objects more than other children
0.03
−0.07
0.09
0.05
0.38
−0.13
0.36
0.20
−0.02
−0.03
MEA
Identify shape in autostereogram [shark]
−0.05
0.00
−0.03
0.01
−0.00 0.91
0.04
0.01
0.01
0.02
MEA
Identify shape in autostereogram [teapot]
0.05
−0.00
0.01
−0.03
0.04
0.91
0.00
0.06
−0.06
−0.03
MEA
Ease of perceiving shapes in
autostereograms above
0.03
−0.02
0.03
0.04
0.01
0.85
0.01
−0.03
0.06
0.04
MEA
Previous successful completion of
autostereogram
−0.09
0.10
0.09
−0.01
−0.01 0.35
0.22
−0.15
0.04
−0.04
ADC
If do sport, likely to be on your own
−0.16
−0.01
0.04
0.05
−0.04 0.06
0.63
−0.08
−0.04
−0.03
ADC
Avoid team games/sports
−0.18
0.11
0.05
0.02
0.00
0.09
0.60
0.03
0.02
0.08
ADC
Difficulties playing team games as child
−0.04
0.13
−0.01
0.14
0.02
0.01
0.47
0.17
−0.05
0.12
ADC
Others commented on clumsiness as child
0.13
0.02
0.06
0.01
0.35
−0.16
0.44
0.23
−0.04
−0.06
ADC
Difficulties with hobbies requiring good coordination
−0.03
0.10
−0.10
0.02
0.09
−0.03
0.25
0.57
0.03
0.05
ADC
Difficulties eating with utensils
−0.03
−0.08
0.06
0.08
0.01
0.05
−0.09
0.57
−0.09
0.06
ADC
Self-care difficulties
−0.08
0.02
0.05
0.19
0.07
−0.08
−0.01
0.48
−0.08
0.08
ADC
Avoid hobbies that require good coordination
−0.02
0.13
−0.08
0.02
−0.02 −0.02
0.36
0.45
−0.03
0.15
AQ
Can easily imagine what characters in
story look like
0.00
−0.01
0.08
−0.16
−0.08 0.08
−0.00
0.05
0.52
−0.00
AQ
Easily play games with children involving pretending
0.28
−0.07
−0.05
−0.10
0.05
0.05
0.08
−0.15
0.43
0.14
AQ
Easy to create a picture using imagination
0.01
−0.01
0.25
−0.03
−0.08 0.08
−0.09
−0.01
0.42
0.02
AQ
Is a good diplomat
0.28
0.02
−0.00
0.05
−0.08 −0.00
−0.11
−0.01
0.37
−0.05
AQ
Making up stories is easy
0.06
0.00
0.23
−0.08
0.08
0.05
0.12
−0.08
0.37
−0.02
ADC
Difficulty performing concurrent tasks
−0.11
0.04
0.02
0.20
0.10
−0.02
−0.05
0.24
0.08
0.41
ADC
Difficulty with distance estimation
0.06
0.12
0.02
−0.06
0.14
−0.07
0.23
0.13
−0.04
0.39
AQ
Easy to do more than one thing at once
0.28
0.04
0.04
−0.05
−0.02 0.02
0.03
−0.09
0.13
−0.34
ADC
Difficulty with navigation
0.00
0.09
−0.09
0.09
0.03
−0.07
0.18
0.16
−0.06
0.32
ADC
Difficulty packing suitcase to go away
0.03
−0.05
0.13
0.09
0.21
0.00
−0.05
0.26
−0.02
0.29
ADC
Difficulty learning to ride bike as child
0.06
0.05
0.04
0.12
0.04
−0.03
0.25
0.13
0.02
0.28
ADC
Difficulty preparing meal from scratch
−0.03
−0.02
0.14
−0.01
0.04
−0.05
−0.09
0.27
−0.16
0.25
AQ
Prefer to do things the same way over and over
−0.05
0.00
0.30
0.16
−0.10 −0.06
0.11
0.02
0.04
0.25
AQ
Know if someone listening to me is getting bored
0.27
−0.05
−0.10
0.03
−0.05 −0.04
−0.07
−0.07
0.29
−0.23
ADC
Difficulties with self-care when child
0.10
−0.03
0.01
0.18
0.15
−0.12
0.18
0.26
0.07
0.22
ADC
Difficulty folding and putting away clothes
0.11
0.07
0.13
0.28
0.24
0.00
−0.01
0.20
−0.03
0.22
AQ
Not upset if daily routine is disturbed
0.27
−0.12
−0.06
0.05
0.06
0.04
−0.05
−0.04
0.04
−0.21
AQ
Enjoy doing things spontaneously
0.29
−0.16
−0.09
0.08
0.15
0.10
−0.13
−0.18
0.06
−0.21 Page 9 of 15 Page 9 of 15 Smith et al. Mediation Loadings below .32 (which explain less than 10% of the variance in that item) are not highlighted and are considered to be
negligible loadings for the purposes of analysis (Continued) Loadings below .32 (which explain less than 10% of the variance in that item) are not highlighted and are considered to be
negligible loadings for the purposes of analysis (Continued)
Measure Item
Social
Stereo Detail Fine motor Org
Magic Eye Isolation Coord Imagine Multi
AQ
Quickly go back to previous activity after interruption
0.19
−0.04
−0.01
0.04
−0.10 0.09
0.08
−0.01
0.23
−0.21
ADC
Slower at getting ready
−0.00
0.07
0.08
0.17
0.28
0.07
−0.03
0.12
−0.13
0.20
AQ
When younger, enjoyed pretend games
with others
0.10
0.04
−0.25
−0.08
0.08
0.10
0.03
−0.20
0.29
0.19
AQ
Carefully plan any activities participated in
−0.13
0.06
0.31
0.07
−0.27 −0.07
0.09
0.14
0.16
0.19
SSI
Book too close to eyes when reading
−0.09
0.29
0.03
−0.09
0.09
−0.05
0.18
0.09
−0.09
−0.18
AQ
Not very good at remembering phone numbers
−0.01
0.04
−0.24
0.19
−0.08 −0.03
0.14
0.01
0.11
0.18
SSI
Experience temporary loss of vision
−0.05
0.16
0.03
0.04
0.13
−0.15
0.00
0.30
0.10
−0.17
AQ
Not good at remembering people’s date of birth
−0.09
−0.02
−0.15
0.20
0.11
0.04
0.12
−0.17
0.09
0.17
AQ
Don’t enjoy reading fiction
0.10
−0.02
0.07
0.03
0.02
0.04
0.06
−0.01
−0.06
0.15
AQ
Rather go to library than a party
−0.24
0.12
0.22
0.08
−0.05 0.04
0.25
0.09
0.08
−0.13
AQ
Concentrate on whole rather than parts
0.13
−0.04
−0.23
−0.05
0.09
0.02
0.00
0.07
0.11
−0.11
ADC
Do you avoid going to clubs/dancing
−0.30
0.09
0.15
0.08
0.06
0.01
0.25
−0.06
−0.04
−0.06
AQ
Last to understand the point of a joke
0.14
0.00
0.32
0.01
0.05
−0.14
0.09
0.16
−0.12
0.05
AQ
Fascinated by dates
0.07
0.03
0.26
−0.02
−0.06 0.10
−0.03
0.14
−0.03
−0.05
AQ
Don’t notice small changes
0.01
0.12
0.02
0.20
−0.02 −0.00
0.11
−0.13
−0.20
0.03
AQ
Rather go to the theater than to a museum
0.29
0.01
−0.09
−0.10
−0.05 −0.05
−0.18
0.18
0.04
0.03
ADC
Difficulties playing music instrument when child
0.04
−0.03
−0.03
0.21
0.18
−0.17
0.18
0.15
−0.01
0.01
SSI
Difference between items 8 and 9
0.01
0.05
0.08
−0.01
−0.04 −0.10
0.04
0.03
0.02
−0.01
SSI
Eyes feel ‘tired’
0.09
0.30
−0.02
0.10
0.06
−0.07
−0.03
0.25
0.04
−0.00 coefficients are presented in Fig. Discussion Table 3 Spearman correlation coefficients for factor scores
extracted using confirmatory factor analysis (CFA)
Stereo
Magic Eye
Social
Isolation
Coord
Magic Eye
0.14*
–
Social
−0.34***
0.05
–
Isolation
0.21**
−0.12
−0.53***
–
Coord
0.20**
−0.18**
−0.55***
0.86***
–
Fine motor
0.19**
−0.08
−0.54***
0.64***
0.67***
Significant relationships are indicated by asterisks. *p < 0.05, **p < 0.01, ***p < .001. Significance values Bonferroni corrected in order to adjust for multiple comparisons Table 3 Spearman correlation coefficients for factor scores
extracted using confirmatory factor analysis (CFA) Table 3 Spearman correlation coefficients for factor
extracted using confirmatory factor analysis (CFA) The aim of the present study was to explore the rela-
tionship between stereopsis, motor ability, and social
skills in a sample of adults. The current research builds
upon prior work by investigating whether the impact of
motor impairment upon social functioning persists in
adulthood, as well as incorporating a variable, stereopsis,
which may underlie deficits in motor ability and thus
have an impact upon social skill. The results indicated
that impaired stereopsis both directly and indirectly af-
fected social skills, in the latter case through mediation Page 10 of 15 Smith et al. Molecular Autism (2018) 9:55 Smith et al. Molecular Autism Fig. 2 Path model with standardised estimates, created as an amalgamation of the mediation analyses. Paths with solid arrows signify a significant
predictive relationship, whereas dashed arrows indicate a non-significant relationship Fig. 2 Path model with standardised estimates, created as an amalgamation of the mediation analyses. Paths with solid arrows signify a significant
predictive relationship, whereas dashed arrows indicate a non-significant relationship younger than the groups surveyed by Kuang et al. [23]
and Cao and Markowitz [90], with 92.6% of the partici-
pants who disclosed their age being under 60 years old,
thus, here we extend the finding of a relationship be-
tween stereopsis and daily living skills to younger and
middle-aged adult populations. by coordination and fine motor skill. Additionally, both
fine motor skill and coordination fully mediated the rela-
tionship between stereopsis and isolation due to motor
proficiency, with coordination explaining much larger
proportion of variance. However, in the full model, isola-
tion due to motor proficiency did not have a significant
relationship with social skills. Whilst there was a relationship between stereopsis and
both types of motor proficiency, the size of this effect
was small within the context of the path model. Discussion A much
stronger association was present between fine motor
skill/coordination and isolation. Of these two facets of
motor skill that showed links with isolation, it was co-
ordination/daily living skills (which require gross motor
ability) that exhibited the largest amount of mediation
between stereopsis and isolation. Whilst there is already
evidence that motor ability correlates with feelings of
isolation and social standing with peers [8–10, 40, 91],
these studies do not tend to differentiate between fine
and gross motor skill. Future work might look at
whether social isolation is due to simple impairment in
gross motor skills or if it might be more specifically at-
tributed to a reduction in daily living skills; such know-
ledge would allow more targeted treatment (such as
physical therapy for gross motor skills versus occupa-
tional therapy for daily living skills). Overall, the results of this study suggest that stereopsis
impairment can affect both motor skill proficiency and so-
cial skills. Additionally, as the final aggregate path model
was a good fit for the data, preliminary support is pro-
vided for the validity of the causal pathways in the model. Associations between stereopsis, motor skills, and isolation
The findings reported here support the hypothesis of
links between impaired stereopsis and both fine and
gross motor skills. In the current study, there was also a
relationship between stereopsis impairment and coord-
ination/daily living skills. Little previous research has
looked at this more functional consequence of impaired
stereopsis. It has been observed that the sensation of
depth afforded by binocular viewing is important for
certain gross motor skills, such as obstacle avoidance
whilst walking [18] and intercepting thrown objects [89],
but only two studies have specifically looked at the con-
tribution of reduced stereopsis to daily living skills. he impact of impaired stereopsis on social skills In a group of older individuals (aged 65 years), Kuang,
Hsu, Chou, Tsai, and Chou [23] found no effect of stere-
opsis on daily living tasks such as cooking and writing,
but they did observe that those with poor stereopsis ex-
hibited a reduction in reported energy/vitality, suggest-
ing that more effort may be required to accomplish daily
living tasks. Cao and Markowitz [90] noted that in a
group of older subjects (aged 50 years) with age-related
macular degeneration, those with reduced stereopsis ex-
perienced difficulty with visual motor skills required for
daily living. The observers in the current study were p
p
p
Impaired stereopsis may affect social skill by causing a re-
duction in general motor ability. The current results are
consistent with those who have previously found an asso-
ciation between motor proficiency and social competence
[8, 34–37, 92, 93]. Whilst fine motor skill and coordin-
ation did mediate the relationship between stereopsis and
social skill, this effect was only partial (the mediation
model accounted for around a third of the variance in the
relationship between stereopsis and social skill). Fine
motor skill, coordination, and stereopsis all exhibited a Page 11 of 15 Page 11 of 15 Page 11 of 15 Smith et al. Molecular Autism (2018) 9:55 Smith et al. Molecular Autism stereopsis reduces the opportunity to develop social skills,
especially in childhood, and this extends into adulthood. This explanation must also be viewed with caution, not
least because it is not clear whether the deficits in those
with strabismus are due to the condition itself or the treat-
ment [98]. Further work to test the whether the predictions
of clinical models extend to the general population is
necessary. similar strength of effect in their relationship with social
skill. That the mediators between stereopsis and social
skill accounted for only a small amount of variance sug-
gests that there are other unmeasured factors that play a
part in the relationship between impaired stereopsis and
reduced social skill. The findings here suggest that stere-
opsis may prove useful in other, as yet unexplored, do-
mains related to social interaction—for instance, the
estimation of interpersonal distance. It is interesting to speculate on the underlying mecha-
nisms between impaired stereopsis and social skills. The-
ories from autism research have attempted to link visual
and social abilities, via a common, generalised, cause [94]. Isolation due to motor proficiency does not predict general
social ability In contrast to previous research which has established
that perceived and/or actual social isolation causes indi-
viduals to change their behaviour and have lower-quality
social interactions [99–101], we did not find that isola-
tion due to motor proficiency significantly predicted so-
cial skill in the full path model. It is likely that motor
ability (represented by the fine motor skill and coordin-
ation variables) is responsible for this relationship, espe-
cially considering the items that constitute the isolation
factor all relate to motor proficiency, specifically in the
context of sport and team games. When isolation is
characterised more fully, including indicators such as so-
cial network size, participation in a range of social activ-
ities (not just those that require motor proficiency), and
perceived lack of social support, the relationship be-
tween isolation and social ability is likely to hold true. he impact of impaired stereopsis on social skills Pellicano and Burr [95] use a Bayesian framework to argue
that flattened priors may account for the changes in aut-
ism. They argue that many of the traits underlying autism
are related to a failure to update perception from prior ex-
perience. It is not clear whether this general deficit ex-
tends to depth and stereo-disparity processing, however,
since people with and without autism integrate depth cues
similarly [51]. An alternative theory proposes that autistic
individuals have enhanced perceptual function (EPF) [96]
in early associative areas of sensory processing (e.g. visual
discrimination), resulting in greater locally oriented pro-
cessing. This account suggests that higher-order process-
ing is not always engaged or mandatory in autism, when a
task can be carried out using lower-level perceptual pro-
cessing. Therefore, when presented with complex and fast
moving social stimuli (e.g. a person speaking), a strong
focus on low-level perceptual features may result in infor-
mation overload and an inability to attend to the relevant
visual cues. This account appears to conflict with the
current results in that we find a link between impaired, ra-
ther than enhanced, perceptual function and social isola-
tion. It is worth noting, however, that the perceptual losses
described here are likely to predominantly come from is-
sues at the earliest stages of perceptual processing such as
lack of eye alignment (strabismus or squint) as well as,
possibly, more neurological deficits. The links proposed
between EPF and social abilities are usually described as
more complex cognitive biases which are not necessarily
linked to depth perception [51]. It is important, therefore,
to consider the impact of both peripheral perceptual and
cognitive differences to understand social behaviour. Limitations It is assumed that the greater correlations between the
AQ and ADC scores compared to the SSI score, and the
motor (fine motor and coordination) and social skills
factor scores compared to the stereopsis factor score re-
flect a greater interdependence of social and motor skills
in development. However, it is possible that the stronger
correlation may be an artefact of the questionnaires
used, with the two questionnaires with the largest
number of questions and covering a range of domains
(the AQ and ADC) correlating most strongly. Coren and
Hakstian [56] have established that whilst the SSI has a
relatively high specificity, the sensitivity is relatively poor
(59.7%). A lab- or clinic-derived measure of stereoacuity
might highlight relatively larger (or smaller, dependent
on whether the stereopsis factor extracted in the current
study actually measures this function) correlations with
social and motor skills. Related to this point, whilst
self-report questionnaires are easy to administer to a
large number of individuals, their subjective nature may
result in biased responses [102]. However, the question-
naires used in this study are well standardised and have
demonstrable construct validity. The ADC and AQ in
particular are commonly employed as research and
screening tools. An alternative explanation for the link between stereopsis
and social abilities and behaviours is that the link is envir-
onmentally mediated and is due to selective reinforcement
of behaviours. As discussed in the introduction, stereopsis
cues to depth are most useful in peri-personal space [20]
and thus would be useful for judging social distance and
interpersonal space. Furthermore, optical conditions that
impair depth perception, such as amblyopia or strabismus,
have also been linked to social exclusion and reduced qual-
ity of life measures [21, 97]. Under this explanation, poor Page 12 of 15 Page 12 of 15 Smith et al. Molecular Autism (2018) 9:55 Smith et al. Molecular Autism (2018) 9:55 Smith et al. Molecular Autism (2018) 9:55 sufficient variability in the data to allow us to conduct
our analyses. It is interesting to note that our correlations between
stereoacuity and Magic Eye proficiency factor scores were
relatively low, although significant (see Table 3); there was
no significant correlation between the ASA and SSI total
scores. For the factor scores, our correlation value is
slightly lower than the value of 0.34 previously found by
Wilmer and Backus [59] in a similar comparison. Clinical implications The first and most important clinical implication of this
study is that visual deficits such as reduced stereopsis
can have far reaching implications on behaviour. Inter-
ventions to improve stereopsis itself have had limited
success but are probably not sufficiently developed to be
recommended to ameliorate the issues described here
[110–112]. Nevertheless, it would be important to ad-
dress the sensory and motor issues in the clinic. Stereop-
sis is not the only cue to depth; many other cues to
depth such as texture gradient, size, occlusion are avail-
able. Those with reduced stereopsis are likely to use cues
differently to those with good or normal stereopsis [51]. For many tasks, simply adding a pattern to a surface can
improve the ability to judge and use depth cues, by pro-
viding more size and texture gradient cues. This can im-
prove activities such as walking and stepping [113, 114]
so may have implications for problems of dexterity and
clumsiness. For social skills, it is possible that therapies
which guide those with reduced stereopsis to use alter-
native cues to judge critical distances such as interper-
sonal
distance
might
be
particularly
effective. For
instance, training people to use rules such as keeping an
arm’s length away rather than relying on implicit cues
might be helpful. Finally, it is possible that the link be-
tween stereopsis and social skill is because the percep-
tual deficits reduce the likelihood that people engage in
social activities. Thus, in this case, it would be the clini-
cian’s role to support the child (or adult) to find social
activities which are not affected by a loss of stereopsis. Our sample was non-stratified and was biased towards
university students; however, we note that the number
of participants sampled was markedly higher than the
majority of studies, which administer the AQ in a non-
clinical sample (which is by far the most commonly re-
ported questionnaire of the ones used in the current
study [104]). The recruitment strategies used for the
current study are similar to the trends noted for other
research involving the AQ, including part of the partici-
pant sample being drawn from participant databases
maintained by universities, and the use of online survey
tools to reach a broader audience [104]. Limitations Our
comparison was slightly different to that previous study in
that we asked people to report their difficulty resolving the
autostereogram image, which could account for some of
the difference. Furthermore, to be successful with the auto-
stereogram, participants require good near convergence
which is not covered by the SSI stereoacuity measure
[103]. As above, conclusions regarding stereoacuity based
on questionnaires must be cautious until they are followed
up with controlled clinic or laboratory measurement. Clinical implications There was a relatively high proportion of participants
who surpassed the threshold for clinically significant
levels of impairment across all of the standardised ques-
tionnaires we used. This may be due to self-selection
bias as the study was advertised as a “survey on correla-
tions between visual ability, coordination, and autistic
traits”. Individuals who perceived themselves as clumsy,
having poor social skills, or problems with visual percep-
tion may have been more likely to take part, creating an
opportunistic selection bias. The particularly high pro-
portion of participants meeting or exceeding the AQ
cut-off may reflect the large proportion of individuals ei-
ther pursuing a STEM degree or in a STEM career, who
are more likely to score higher on the AQ than those in
non-STEM education or career paths [105]. Further-
more, whilst only six participants disclosed a diagnosis
of autism or Asperger’s syndrome, more specified that
they were first-degree relatives of someone with the con-
dition. It is thought that autistic traits may be expressed
to a greater degree in close relatives of people with an
ASD, even though they might not meet the criteria for
clinical diagnosis [106], a concept termed the broader
autism phenotype [107–109]. However, our final sample
showed a broad range of individual differences in the
scores of the SSI, AQ, and ADC, indicating that whilst
self-selection bias may have occurred, there was still Conclusions This study has demonstrated the presence of a relation-
ship between stereopsis, motor ability, and social skill. Using a large group of adults, this work complements re-
search previously conducted with children, in addition to
providing evidence for an underlying contributor to im-
pairment in both motor and social skill. Preliminary sup-
port for causal pathways between stereopsis, motor ability,
and social skill has been provided, but further evidence is
needed to clarify the mechanisms responsible, especially
in clinical populations. The repercussions of poor stereop-
sis have been demonstrated to be far-reaching, limiting
not only motor skill, but also social competence. Funding
Th
k 11. Grant S, Melmoth DR, Morgan MJ, Finlay AL. Prehension deficits in
amblyopia. Investig Opthalmology Vis Sci. 2007;48:1139–48. This work was supported by the Economic and Social Research Council
[grant number ES/J500100/1], by a PhD studentship to DS. The funders had
no role in the study design, data collection, analysis, decision to publish, or
preparation of the manuscript. 12. Melmoth DR, Finlay AL, Morgan MJ, Grant S. Grasping deficits and
adaptations in adults with stereo vision losses. Invest Ophthalmol Vis Sci. 2009;50:3711–20. https://doi.org/10.1167/iovs.08-3229. 13. Niechwiej-Szwedo E, Goltz HC, Chandrakumar M, Wong AMF. The effect of
sensory uncertainty due to amblyopia (lazy eye) on the planning and
execution of visually-guided 3D reaching movements. PLoS One. 2012;7:
e31075. Received: 4 December 2017 Accepted: 21 September 2018 Additional file 2: Table S2. The psychiatric or organic illnesses self-
disclosed in the feedback section of the questionnaire. Note that diagnoses
were not collected routinely as part of the demographical data. The data
below represent a number of co-morbidities: 32 diagnoses were disclosed
by 24 participants (3.7% of the sample). (DOCX 24 kb) Availability of data and materials The dataset generated and analysed during the current study is available in
the Figshare repository, DOI: https://doi.org/10.6084/m9.figshare.5364220 The dataset generated and analysed during the current study is available in
the Figshare repository, DOI: https://doi.org/10.6084/m9.figshare.5364220 14. Schiller PH, Kendall GL, Kwak MC, Slocum WM. Depth perception, binocular
integration and hand-eye coordination in intact and stereo impaired
human subjects. J Clin Exp Ophthalmol. 2012;3:1–12. Authors’ contributions All authors developed the study concept and design. DS created the survey
website, recruited participants, analysed the data, and wrote the first draft of
the article, under supervision of HA and DR. All authors contributed to the
final paper and approved the final version for submission. 15. Grant S, Suttle C, Melmoth DR, Conway ML, Sloper JJ. Age- and
stereovision-dependent eye-hand coordination deficits in children with
amblyopia and abnormal binocularity. Invest Ophthalmol Vis Sci. 2014;55:
5687–57015. Additional files Additional files Additional file 1: Table S1. A detailed breakdown of self-reported
occupation, including faculty for those in education (where available) and
sector for those in employment. (DOCX 25 kb) Page 13 of 15 (2018) 9:55 Smith et al. Molecular Autism (2018) 9:55 Smith et al. Molecular Autism Received: 4 December 2017 Accepted: 21 September 2018 Received: 4 December 2017 Accepted: 21 September 2018 References Transition from crawling to
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maps and institutional affiliations. Abbreviations
3D Th
di 3D: Three-dimensional; ADC: Adult Developmental Coordination Disorder
Checklist; AQ: Autism Spectrum Quotient; ASA: Autostereogram Self-Assessment;
ASD: Autism spectrum disorder; CFA: Confirmatory factor analysis;
CFI: Comparative Fit Index; DCD: Developmental coordination disorder;
EFA: Exploratory factor analysis; MCAR: Missing completely at random;
RMSEA: Root mean square error of approximation; SEM: Structural equation
modelling; SRMR: Standardised root mean square residual; SSI: Stereopsis
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Link Scheduling in Rechargeable Wireless Sensor Networks With Imperfect Batteries
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Tony Tony
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Received July 11, 2019, accepted July 25, 2019, date of publication July 30, 2019, date of current version August 14, 2019. Received July 11, 2019, accepted July 25, 2019, date of publication July 30, 2019, date of current version August 14, 2019. Received July 11, 2019, accepted July 25, 2019, date of publication July 30, 2019, date of current version August 14, 2019. Digital Object Identifier 10.1109/ACCESS.2019.2932079 The associate editor coordinating the review of this manuscript and
approving it for publication was Yi Zhang. INDEX TERMS Link scheduling, wireless sensor networks, TDMA, HUS, harvesting time, battery capacity,
battery leakage, storage efficiency. INDEX TERMS Link scheduling, wireless sensor networks, TDMA, HUS, harvesting time, battery capacity,
battery leakage, storage efficiency. INDEX TERMS Link scheduling, wireless sensor networks, TDMA, HUS, harvesting time, battery capacity,
battery leakage, storage efficiency. his work is licensed under a Creative Commons Attribution 4.0 License. For more information, see http://creativecommons.org/licenses/by/4.0/ TONY TONY
1, (Member, IEEE), SIETENG SOH1, (Member, IEEE),
KWAN-WU CHIN
2, AND MIHAI LAZARESCU1, (Member, IEEE)
1School of Electrical Engineering, Computing, and Mathematical Sciences, Curtin University, Perth, WA 6102, Australia
2School of Electrical, Computer, and Telecommunications Engineering, University of Wollongong, Wollongong, NSW 2500, Australia
Corresponding author: Tony Tony (tony@postgrad.curtin.edu.au) TONY TONY
1, (Member, IEEE), SIETENG SOH1, (Member, IEEE),
KWAN-WU CHIN
2, AND MIHAI LAZARESCU1, (Member, IEEE)
1School of Electrical Engineering, Computing, and Mathematical Sciences, Curtin University, Perth, WA 6102, Australia
2School of Electrical, Computer, and Telecommunications Engineering, University of Wollongong, Wollongong, NSW 2500, Australia
Corresponding author: Tony Tony (tony@postgrad.curtin.edu.au) This work was supported in part by the Indonesia Lecturer Scholarship (BUDI) from the Indonesia Endowment Fund for
Education (LPDP), Ministry of Finance, Indonesia. ABSTRACT This paper considers the problem of generating the shortest time division multiple access
(TDMA) schedule for use in rechargeable wireless sensor networks (rWSNs) with heterogeneous energy
arrivals rates. This novel problem considers: 1) the time required by nodes to harvest sufficient energy to
transmit/receive a packet; 2) harvest-use-store (HUS) energy harvesting and usage models, and; 3) battery
imperfections, i.e., leakage, storage efficiency, and capacity. This paper shows the problem at hand, called
link scheduling in harvest-use-store (LSHUS), is in general NP-Complete. Furthermore, it presents a greedy
heuristic, called LS-rWSN, to solve LSHUS. Our experiments show that a longer energy harvesting time
(leakage rate) from 1 to 20 (0% to 4%) increases the schedule length by up to 565.82 (44.54%) slots while
reducing storage efficiency from 1.0 to 0.6 lengthens the schedule by up to 62.77%. In contrast, battery
capacity has an insignificant effect, i.e., enlarging the capacity by 20 times decreases the schedule length by
only 6.5%. I. INTRODUCTION Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries FIGURE 1. An example (a) with only protocol interference constraint, and
(b) protocol interference, harvesting time, and battery capacity
constraints. The number next to each link denotes its activation time, and
vx |z denotes node x requires z time slots to recharge its battery to a level
before the next transmission or reception is possible. is only sufficient to store the energy required to receive one
packet. Consequently, the battery of node v2 can be recharged
only after it is used at time t = 5, and it is fully recharged at
time t = 5 + 2 = 7. Thus, node v2 can receive the second
and third packet no earlier than at time t = 7 and t = 9,
respectively, e.g., link (v3, v2) can be scheduled at time t = 7,
and link (v4, v2) at time t = 9, which yield a schedule of
length nine; see Fig. 1b. This example shows that battery
capacity affects the schedule length. FIGURE 1. An example (a) with only protocol interference constraint, and
(b) protocol interference, harvesting time, and battery capacity
constraints. The number next to each link denotes its activation time, and
vx |z denotes node x requires z time slots to recharge its battery to a level
before the next transmission or reception is possible. Two additional factors that can affect the schedule length
are battery leakage and storage efficiency. The typical value
of storage efficiency can be as low as 66% [11] depending on
the battery technology. Similarly, the leakage rate of a battery
depends on its type, age, usage, and/or temperature [12]. As an example, the leakage rate of a Li-ion battery is 8%
per month [13], which is lower than a Nickel-based battery. Further, the leakage rate changes over times and has the
highest leakage right after being charged [12]. To illustrate
the effect of these factors on the link schedule, reconsider the
previous example depicted in Fig. 1b. Assume sensor nodes
have a battery with a capacity of one unit of energy (1ϵ). Now
assume the battery of nodes leaks at a rate of 5% per slot, and
has a storage efficiency of 90%. Given this imperfect battery,
the schedule length becomes 12 slots instead of 9 slots. I. INTRODUCTION This
is because links (v1, v2), (v3, v2), and (v4, v2) are scheduled
at time t = 6, t = 9, and t = 12, respectively. The reason is
because nodes v1, v2, and v3 require more than five, six and
seven time slots respectively to accumulate sufficient energy
to transmit or receive a packet due to battery leakage and
storage efficiency. listening [9]. Hence, a link scheduler plays a critical role in
a rWSN. Past works on link scheduling assume nodes have no
energy constraints [10]. In contrast, in a rWSN, link sched-
ulers must consider the varying energy harvesting rates of
sensor nodes. Specifically, they must consider the energy
harvesting time of nodes; i.e., this is the time interval in
which a sensor node accumulates sufficient energy to either
transmit or receive a packet. Without this consideration, a link
scheduler may allocate slots to nodes that have insufficient
energy to transmit/receive. Another important issue to con-
sider is battery characteristics; namely (i) limited capacity
aka battery capacity, (ii) leakage, and (iii) storage efficiency. These characteristics can result in a longer link schedule. We now discuss the aforementioned issues with the aid
of an example. Fig. 1a and 1b show two rWSN examples
with four nodes and three directed links; the number next
to each link refers to its activation time slot. Links (v1, v2),
(v3, v2), and (v4, v2) interfere with each other and thus cannot
be scheduled to transmit concurrently. Assume links (v1, v2),
(v3, v2) and (v4, v2) are to be activated in time slot t = 1,
t = 2, and t = 3, respectively. This means the resulting
TDMA schedule or superframe is three slots in length. The
key assumption for the example in Fig. 1a is that nodes
have sufficient energy to transmit and receive in its allocated
time slot. Next, consider the case where sensor nodes have a
different energy harvesting cycle. Fig. 1b shows that node v1
is able to transmit/receive every five slots; denoted as v1|5. This causes the schedule length to exceed three slots as each
node must now wait for its battery to recharge. Notice that
node v2 has sufficient energy at time slot t = 2. However,
none of its incoming links can be activated at time 2 because
its neighbors have insufficient energy to transmit a packet. I. INTRODUCTION Specifically, link (v1, v2) can be scheduled no earlier than slot
t = 5 because node v1 can only transmit after time t = 5 This paper contains the following contributions: p p
g
• It proposes a novel TDMA link scheduling problem,
called Link Scheduling in Harvest-Use-Store (LSHUS),
to maximize the throughput of rWSNs whereby (i) sen-
sor nodes have a different energy harvesting cycle,
(ii) sensor nodes have a battery with finite capacity,
and each battery has less than ideal storage efficiency
and leaks over time, and (iii) each link i has a weight
wi ≥1 that specifies that it must be scheduled at least wi
times in the resulting schedule. To the best of our knowl-
edge, no link schedulers have simultaneously considered
factors (i)-(iii). The authors of [14] consider factors
(i) and (iii) and they assume nodes use the Harvest-
Store-Use (HSU) model with unlimited battery capacity. A prior work in [15] uses the Harvest-Use-Store (HUS)
model and considers batteries with limited capacity. More specifically, the work in [14] and [15] assumes
batteries that are leakage free and have 100% storage
efficiency. This paper extends the work in [15] by also
considering different leakage rates and storage efficien-
cies. Note, this paper only considers the HUS model as
it is superior to the HSU model; see [15] for details. • It proposes a novel TDMA link scheduling problem,
called Link Scheduling in Harvest-Use-Store (LSHUS),
to maximize the throughput of rWSNs whereby (i) sen-
sor nodes have a different energy harvesting cycle,
(ii) sensor nodes have a battery with finite capacity,
and each battery has less than ideal storage efficiency
and leaks over time, and (iii) each link i has a weight
wi ≥1 that specifies that it must be scheduled at least wi
times in the resulting schedule. To the best of our knowl-
edge, no link schedulers have simultaneously considered
factors (i)-(iii). The authors of [14] consider factors
(i) and (iii) and they assume nodes use the Harvest-
Store-Use (HSU) model with unlimited battery capacity. A prior work in [15] uses the Harvest-Use-Store (HUS)
model and considers batteries with limited capacity. More specifically, the work in [14] and [15] assumes
batteries that are leakage free and have 100% storage
efficiency. This paper extends the work in [15] by also
considering different leakage rates and storage efficien-
cies. I. INTRODUCTION to four seconds, aka harvesting/recharging times or cycles,
before it can transmit or receive one packet. Wireless Sensor Networks (WSNs) will play a crucial role
in the Internet of Things (IoTs) era [1]. In particular, WSNs
will be critical to information/data gathering of activities in
environments such as smart homes [2] or to monitor the blood
pressure of a person [3]. A key concern when operating a
WSN, however, is managing the ambient energy harvested
by sensor nodes [4]. Of interest, is ensuring sensor nodes
used stored energy efficiently. Moreover, once a node’s bat-
tery is full, any subsequent energy arrivals are loss. Also,
the energy harvesting rate is location specific [5]. Once a
sensor node exhausts its harvested energy, it will have to
spend time accumulating energy before it is able to carry
out any tasks. For instance, the energy generated by air-flow
has a power density of 360 µW/cm2 [6]. With a surface area
of 50 cm2, its energy harvesting rate is 18 mJ/s. If the har-
vester powers a Mica2 mote [7] that works with 72 mJ of
energy to transmit/receive a packet, the mote has to wait up In a rechargeable wireless sensor network (rWSN), sensor
nodes need to access the channel in order to transmit/receive
packets. To this end, this paper addresses the problem of
deriving a short Time Division Multiple Access (TDMA)
schedule for use in a rWSN. Such a schedule ensures
collision-free transmissions, meaning energy is not wasted
due to collisions. Specifically, a link scheduler is responsible
for determining the set of transmitting and receiving nodes in
each time slot. As the TDMA schedule repeats, it is important
that the schedule length (in terms of slots) is short as this
allows nodes to transmit frequently; consequently, as links
are activated frequently, they will have a high capacity. It is
preferable to have a link schedule to be as short as possible
and for each transmission slot to have as many links as
possible. Hence, as the link schedule repeats periodically,
such a schedule will result in a high network capacity [8]. Lastly, the schedule governs the active time of a node, mean-
ing a node only needs to become active if its neighbors
are active. In other words, such a schedule minimizes idle The associate editor coordinating the review of this manuscript and
approving it for publication was Yi Zhang. 104721 T. II. RELATED WORKS To the best of our knowledge, except for reference [14] and
our prior work [15], there are no works that solve a similar
problem to ours. Liu et al. [16] aim to optimize the throughput
and transmission time in many-to-one networks . The authors
consider two cases: (i) infinite, and (ii) finite battery capacity. Kapoor and Pillai [17] aim to find efficient schedulers for
energy harvesting nodes operating over a multiple access
channel. Lenka et al. [18] design a hybrid medium access
control (MAC) protocol for WSNs that minimizes latency
and collisions. He et al. [19] present link scheduling, data
routing and energy sharing in rWSNs to maximize the min-
imum source or sensing rate of nodes. On the other hand,
Li et al. [20] study a scheduling optimization problem for a
Energy Harvesting (EH) mobile WSN. They aim to maximize
the amount of data collected from sensors by scheduling the
transmission per time slot according to the energy harvested
by sensor nodes and link quality. The authors of [21] inves-
tigate a joint data gathering and EH problem in rWSNs with
a mobile sink. The goal is to maximize the network utility by
jointly considering the relay selection, power/energy alloca-
tion and time scheduling problems. However, none of these
works consider recharging time of batteries at the end nodes
of active links. The work in [12], [26], and [11] considers batteries with
leakage and storage efficiency; aka imperfect batteries. The
authors in [12] propose a framework to maximize the amount
of data transmission by adjusting the transmit power in an
energy harvesting system with battery limitation such as leak-
age constraint. Biason and Zorzi [26] proposed a framework
based on a Partially Observable Markov Decision Process
(POMDP). The goal is to optimize the throughput of energy
harvesting-capable devices. Further, they consider the effects
of imperfect batteries. In [11], Tutuncuoglu et al. study two
policies: (i) optimal offline, and (ii) online to maximize the
average transmission rate in an energy harvesting network
with an inefficient finite capacity battery that loses a constant
fraction of its stored energy. However, none of these papers
consider link scheduling. g
In summary, there are no prior works on link schedul-
ing for rWSNs that consider all of the following factors:
battery recharging time, capacity, leakage, and storage effi-
ciency. Sun et al. [14] consider recharging time, and assume
unlimited battery capacity. I. INTRODUCTION Note, this paper only considers the HUS model as
it is superior to the HSU model; see [15] for details. 1
y
The battery capacity of sensor nodes is also a key factor that
affects the schedule length. Consider the case where at time
t = 3, node v2 continues to accumulate energy, and hence
at time t = 5 and t = 7, it has sufficient energy to receive
two and three consecutive packets, respectively. For this case,
links (v1, v2), (v3, v2), and (v4, v2) can be scheduled at time
t = 5, t = 6, and t = 7, respectively, giving a schedule
of length seven. Now assume the battery capacity of node v2 The battery capacity of sensor nodes is also a key factor that
affects the schedule length. Consider the case where at time
t = 3, node v2 continues to accumulate energy, and hence
at time t = 5 and t = 7, it has sufficient energy to receive
two and three consecutive packets, respectively. For this case,
links (v1, v2), (v3, v2), and (v4, v2) can be scheduled at time
t = 5, t = 6, and t = 7, respectively, giving a schedule
of length seven. Now assume the battery capacity of node v2 • It outlines an efficient greedy technique to generate
TDMA link schedules, and contains analysis of its time
complexity. In addition, it also presents a proof to show
that LSHUS is NP-complete, and contains analysis of 104722 104722 VOLUME 7, 2019 VOLUME 7, 2019 T. Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries the optimal schedule length for the following topologies:
Line, Tree, and Grid. The proposed technique does not
require an extended conflict graph as in [14], and thus
it is more efficient. The results in Section V show that
imperfect batteries increase the schedule length. In par-
ticular, the results show that at a battery leakage rate
of 0.01 and storage efficiency of 0.7, the superframe
length increases by up to 63.68% as compared to a
perfect battery when the leakage rate is at zero and
storage efficiency is at 100%. Further, increasing the
harvesting time from one to 20 lengthens the schedule
by up to 565.82%. The proposed heuristic can produce
schedule lengths that are on average only 1.07 times
longer than the lower bound length. I. INTRODUCTION It also produces the
optimal schedule for the Line topology, and achieves
at most 28% and 42% longer schedule lengths than
the optimal length for the BTree and Grid topologies,
respectively. capacity, 100% storage efficiency, and are leakage free. The
links in [14] can be scheduled only if the battery of their end
nodes have accumulated sufficient energy to transmit/receive
one data packet. Each node with insufficient energy thus must
wait for at least one recharging cycle before it can activate one
link. The authors propose two link schedulers to maximize
network throughput: (i) without link weight (or wi,j = 1),
and (ii) with link weight (wi,j > 1). For (i), they generate
a conflict graph CG(V ′, E′) from G(V, E) according to the
protocol interference model. For (ii), their scheduler requires
an extended conflict graph C′G(V ′, E′′), which is generated
from CG and wi,j of each link such that each link (i, j) appears
wi,j times in C′G. ,j
The HUS protocol is first introduced in [23]. According
to [22], HUS has a higher achievable harvesting rate and
lower energy loss as compared to HSU. Recently, there are
works that consider HUS, but these works do not consider the
problem in this paper. More specifically, in [24], Yuan et al. investigate the HUS architecture for point-to-point data trans-
mission with a rechargeable battery over two channels:
(i) static, and (ii) block fading. They aim to maximize
throughput and propose optimal energy policies based on
a discrete-time energy model. They then extend their work
in [25], whereby the aim is to minimize the energy used for
transmissions subject to a delay constraint. However, these
papers assume perfect battery, i.e. batteries with zero leakage
and 100% storage efficiency. The rest of this paper is organized as follows. Section II
reviews related works, followed by Section III, which con-
tains network model and problem. Section IV describes the
problem and solution. The performance evaluation is reported
in Section V. Finally, Section VI concludes the paper and
provides future research directions. II. RELATED WORKS The authors consider the HSU
model [22]. The HSU model requires the harvested energy
at slot t to be used no earlier than slot t + 1, which results
in a longer schedule length. The work in [15] considers
the HUS protocol [22], which offers better performance. In HUS, nodes can use its harvested energy immediately, and
hence, reduce energy loss and produce shorter link schedule,
meaning larger throughput, than a link schedule that uses the
HSU model. Further, the approach in [15] only requires a
conflict graph, unlike the solution in [14] that also requires an The most relevant works to ours are [14] and [15]. Sun et al. [14] consider the HSU model [22], whereby har-
vested energy must be first stored in a battery before it can
be used. Each battery has a recharging time that determines
when a node has sufficient energy to transmit/receive one
packet. The authors assume nodes have a perfect battery with
unlimited capacity; i.e., nodes use batteries with unlimited 104723 104723 VOLUME 7, 2019 T. Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries TABLE 1. A comparison of prior works. extended conflict graph. This is advantageous as the extended
conflict graph becomes computationally expensive to use
with a large link weight wi,j. However, the work in [15] does
not consider battery leakage and storage efficiency. Note that
a Ni-MH rechargeable battery only can store 70% of the
harvested energy [22]. As a result, some valuable energy is
loss due to energy storage inefficiency and leakage. Hence-
forth, this paper extends the work in [15] to include these
two important factors. A quick comparison of our work and
previous works can be found in Table 1. III. PRELIMINARIES
Section III-A first describes the rWSN model under con-
FIGURE 2. A rWSN model: (a) graph G, and its (b) conflict graph CG. Also
shown are primary (dashed lines) and secondary (solid line) interference BLE 1. A comparison of prior works. TABLE 1. A comparison of prior works. FIGURE 2. A rWSN model: (a) graph G, and its (b) conflict graph CG. Also
shown are primary (dashed lines) and secondary (solid line) interference. extended conflict graph. This is advantageous as the extended
conflict graph becomes computationally expensive to use
with a large link weight wi,j. However, the work in [15] does
not consider battery leakage and storage efficiency. III. PRELIMINARIES Section III-A first describes the rWSN model under con-
sideration. It then introduces key notations (see Table 2). Section III-B then formally presents the problem at hand. This section also shows the said problem is NP-complete. Section III-C presents an analysis of how factors such as har-
vesting time, battery storage efficiency, and leakage impact
the schedule length for fixed topology networks. transmission range of each node vi, and ||vi −vj|| represents
the Euclidean distance between vi and vj. Node vi can trans-
mit or receive packets to/from vj if the condition ||vi −vj|| ≤
Ri is true. Each link li,j has weight wi,j ≥1 that indicates the
number of time slots that it requires in the resulting schedule;
e.g., Fig. 2a shows w1,2 = 3. This paper assumes links are activated as per the protocol
interference model [27]. Specifically, primary interference
occurs when a node transmits and receives a packet simulta-
neously, or receives more than one transmission at the same
time; that is, each node is half-duplex. Secondary interference II. RELATED WORKS Note that
a Ni-MH rechargeable battery only can store 70% of the
harvested energy [22]. As a result, some valuable energy is
loss due to energy storage inefficiency and leakage. Hence-
forth, this paper extends the work in [15] to include these
two important factors. A quick comparison of our work and
previous works can be found in Table 1. FIGURE 2. A rWSN model: (a) graph G, and its (b) conflict graph CG. Also
shown are primary (dashed lines) and secondary (solid line) interference. A. NETWORK MODEL A directed graph G(V, E) is used to model a rWSN, where
each node vi ∈V is a sensor node i and each link li,j ∈E
denotes a directed link from vi to vj. Let Ri be the 104724 VOLUME 7, 2019 VOLUME 7, 2019 T. Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries TABLE 2. Notations and definitions. TABLE 2. Notations and definitions. TABLE 2. Notations and definitions. no sensor nodes have sufficient energy to transmit/receive. Note that prior link schedulers assume nodes always have
energy when they are scheduled to transmit/receive; this
paper relaxes this assumption. A sensor node consumes energy when sensing the environ-
ment, computing collected samples, and communicating with
its neighbors, which include transmitting, receiving, listening
for messages on the radio channel, sleeping, and switching
state [30]. This work assumes that communication is the only
source of energy expenditure. The assumption is reasonable
because as shown in [30], the energy consumption of nodes
for communications is significantly larger than other opera-
tions, e.g., 180.10 mJ, 17.242 mJ, and 5.2 mJ for communi-
cation, sensing, and computing, respectively. Similar to [31],
assume the energy usage for transmission and reception is
equal. Let ϵ (in Joule) be the energy consumed when trans-
mitting or receiving one packet. For example, assuming a TI
CC2420 transceiver that uses 226 nJ/bit for transmission [32],
and a packet size of 125 bytes or 1,000 bits, then we have
ϵ = 226 µJ. µ
This paper considers nodes that use the HUS model [22],
where harvested energy is first stored in a capacitor for imme-
diate use and any unused energy is stored in a rechargeable
battery for use in future slots. Note that this paper does
not make any specific assumption about any energy har-
vesting model; i.e., the problem - to be formally defined in
Section III-B - is independent of any specific energy harvest-
ing model. In particular, it considers the amount of energy that
arrives after energy conversion. Hence, solution to the prob-
lem continues to work if the energy source is solar, which is
linear with respect to the solar panel size, or Radio Frequency
(RF), which is non-linear with respect to the input power. A. NETWORK MODEL To this end, a node i contains a harvester that generates energy
from its environment, e.g., the sun, and two energy storage
types: (i) a super capacitor with a capacity of ci, and (ii) a
rechargeable battery with a capacity of bi; both capacities are
in unit of ϵ. Let ri > 0 (in slots) be the total number of
slots or harvesting time required by a node i to accumulate
1ϵ amount of energy. Thus, a node has a harvesting rate
of ϵ
ri per time slot. Each capacitor for node i is assumed to
have sufficient capacity to store all harvested energy in each
slot, i.e., ci ≥1/ri. The energy level of each capacitor is
zero at the start of each time slot. Any unused energy that
the harvester receives at slot t, e.g., any excess energy when
ri < 1 or node i is not active, is stored in a rechargeable
battery for use in slot t+1 and thereafter. The battery of nodes
is initially empty. occurs when say a node A, while receiving a packet from its
neighbor B, also receives a transmission from node C that is
intended for another node D. In Fig. 2a, there are two primary
interference; i.e., link l4,3 with l3,1, and l3,1 with l1,2. Also
shown is the secondary interference at node v3 that is caused
by node v1. y
The interference between links is aptly modeled by a
conflict graph CG(V ′, E′) [28]. For a given G(V, E), its
corresponding conflict graph can be constructed as follows:
(i) each vertex in V ′ represents a link in E, i.e., |V ′| = |E|, and
(ii) each edge in E′ represents two links of G that experience
primary or secondary interference if they are active together. Fig. 2b shows the conflict graph CG for the rWSN in Fig. 2a. The figure shows primary and secondary interference in a
dashed and a solid line, respectively. Any graph coloring
algorithms, such as edge coloring [29], can then be applied on
the conflict graph to determine the links that can be scheduled
together. That is, all links with the same color do not inter-
fere and thus can transmit together. A. NETWORK MODEL Note that our problem
and its solution, described in Section III-B and Section IV
respectively, can also be used for other interference models,
e.g., the Request to Send/Clear to Send (RTS/CTS)-based
model [14]. A TDMA superframe or a link schedule is defined as a
collection of consecutive, equal sized time slots. All links in
each slot do not experience primary and secondary interfer-
ence. Indeed, after coloring a conflict graph, all links with
the same color can be placed in a slot. Let S represent the
superframe and |S| denote the schedule length (in slots). Each
slot has size τ (in seconds), and is sufficient to transmit one
packet. Without loss of generality, τ is set to one millisecond
(ms). Each slot is either empty or contains one or more non-
interfering, concurrently active links. A slot is empty when Let 0 < ηi ≤1 be the storage efficiency and 0 ≤
µi
< 1 be the battery leakage factor (per time slot) of
node i. Notice that values ηi = 0 and µi = 1 correspond
to the case where the battery cannot store any harvested
energy and retain its charge or energy, respectively, and thus
are considered. Further, in each slot, the amount of energy
that can be stored in a battery from scavenging energy is
larger than the battery’s energy leakage. The requirement is
necessary because otherwise any harvested energy will be lost 104725 104725 VOLUME 7, 2019 T. Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries FIGURE 3. TDMA schedules for rWSN in Fig. 2. Gray colored slots indicate
no transmissions/receptions. immediately due to battery leakage. Consequently, nodes will
not have any energy to operate. The rechargeable battery of nodes supports shallow
recharging [4], meaning it can be recharged even though it
is partially discharged. Let bi,t be the energy level of node i’s
battery at time t. For each node i, Ai,t (in unit of ϵ) represents
the amount of energy that node i is allowed to use in slot t;
note, 1
ri ≤Ai,t ≤bi,t + 1
ri . The value of Ai,t is the sum
of energy level of node i’s battery and capacitor at time t,
that is, Ai,t = bi,t + 1
ri . B. PROBLEM STATEMENT For a given rWSN, the Link Scheduling in Harvest-Use-
Store (LSHUS) problem is to generate a TDMA link sched-
ule S with the shortest length |S| such that (i) each link li,j
that is allocated a time slot t satisfies Ai,t ≥1ϵ and Aj,t ≥1ϵ,
and (ii) each link li,j ∈E is scheduled at least wi,j times in S. For example, in Fig. 2a, link l3,1 can be scheduled no earlier
than t = 5; and link l1,2 needs to be scheduled three times
because w1,2 = 3. To illustrate the effect of link scheduling on |S|, consider
the example in Fig. 2 with equal µi = 0 and ηi = 1. Fig. 3a
shows one feasible schedule. A schedule is called feasible if
it satisfies constraints (i) and (ii). The optimal solution can
be found in Fig. 3b. Note that the figure shows only non
empty slots, i.e., each empty slot is represented as ‘‘...’’. The problem aims to generate a schedule S with the shortest
length, e.g., the schedule in Fig. 3b has length |S| = 18. A battery cannot be charged and discharged simultane-
ously [33]. However, its energy level may increase when a
node uses energy at the same time slot t. This case occurs
when ri < 1. On the other hand, when ri > 1 and bi,t ≥
1 −1
ri , then node i will draw the fraction 1 −1
ri , aka energy
shortfall, from its battery. Finally, since the amount of leaked
energy can never be larger than the energy stored in a node’s
battery, we have that the amount of energy that a node i
uses at time t + 1 is larger or equal to the energy at time t;
i.e., Ai,t+1 ≥Ai,t. Let SE represent the superframe generated when there is no
interference In this case, the activation of links is delayed by
insufficient energy as opposed to interference. One can use
|SE| as lower bound of the superframe length for LSHUS,
computed as Let Ti denote the earliest time slot when node i has at least
1ϵ of energy to transmit/receive one packet. In other words,
Ti is the earliest slot such that Ai,Ti ≥1ϵ. The earliest time
link li,j can be scheduled is thus at time ti,j = max(Ti, Tj). A. NETWORK MODEL As the capacitor has a high leakage
rate [22], the energy level in each capacitor is always equal
to the energy harvested in each slot, i.e.,
1
ri . This paper
considers the capacitor of nodes has 100% energy storage
efficiency. When Ai,t < 1ϵ, node i cannot transmit or receive
packets at time t. In contrast, if ri ≤1 or Ai,t
≥1ϵ,
a node can transmit/receive at any time, assuming there is
an available packet. The available energy Ai,t is a function
of node i’s battery capacity (bi), energy harvesting rate (ri),
storage efficiency (ηi), leakage rate (µi), and energy usage. Let ti be the time in which node i last draws energy from
its battery. When Ai,ti
< 1, it takes ρi
= t −ti slots
for node i to accumulate energy such that it has Ai,t ≥1. As explained later in Section IV-A, ρi is affected by the
harvesting time ri, storage efficiency ηi, leakage rate µi, and
energy level bi,ti. FIGURE 3. TDMA schedules for rWSN in Fig. 2. Gray colored slots indicate
no transmissions/receptions. earliest time node 1 and 2 can transmit or receive is at slot
T1 = 6 + 2 = 8 and T2 = 12, respectively. 1) LINE GRAPH Assume a Line graph with n nodes. The nodes are labeled
consecutively from 1 to n. Following the protocol interference
model, a node cannot transmit and receive at the same time. Further, any nodes within two hops away are not allowed to
transmit in the same slot when at least one of them transmits
to their common neighbor. For example, there is interference
between links (2, 1) and (4, 3), and (2, 3) and (4, 5) at node 3,
but links (2, 1) and (4, 5) are interference free. Proposition 1: The optimal link schedule for Line topol-
ogy with n ≥3 has superframe length |S| = 4wr. Proof: This proof first describes how construct a link
schedule with 4wr time slots, for two cases: r = 1 and r > 1. Then, it shows that the result is optimal. B. PROBLEM STATEMENT Recall that ρi is the number of slots for a node i
to accumulate energy to reach at least 1ϵ worth of energy,
starting from the time the node has Ai,t < 1ϵ. Note, all
three topologies are bipartite graphs [29]. Briefly, a graph is
bipartite if its nodes can be placed into two sets, namely Set-1
and Set-2, with links between nodes in Set-1 and Set-2 only. p y
To show how the solution for LSHUS for instance G′ gives
the solution for ScheduleEV on instance of G, consider a
special case of LSHUS, i.e., each node has harvesting time
ri = 1, and each battery has a capacity of bi = 1, storage
efficiency ηi = 1, and leakage rate µi = 0. For this case, each
node always has sufficient energy to transmit or receive one
packet because ri = 1. Thus, the superframe S1 of LSHUS
for instance G′ is exactly the solution for instance G of
ScheduleEV. Thus, LSHUS is at least as hard as ScheduleEV,
i.e., LSHUS is also an NP-complete problem. It is impor-
tant to note that LSHUS is more complex than ScheduleEV
because it considers any arbitrary topologies, not just tree
topologies in the latter problem. Moreover, LSHUS considers
ri ≥1, bi ≥1, ηi ≤1, and µi ≥0. B. PROBLEM STATEMENT An instance of (a) graph G and its mapping (b) graph G′. FIGURE 5. A line graph with eight nodes. FIGURE 4. An instance of (a) graph G and its mapping (b) graph G′. FIGURE 4. An instance of (a) graph G and its mapping (b) graph G′. FIGURE 6. A binary tree with four levels. LSHUS as follows. Firstly, set G′ = G, i.e., V ′ = V, and E′ =
E. Secondly, for each node i in G′, compute its weight wi
as follows. For each leaf node in G′, set wi = ωi. Then,
compute the weight of each parent node j by summing the
weight of all its children and ωj. Note that one can interpret
each weight wi as the total number of packets that node i in
G′ must transmit to the root node; further, nodes forward their
own packets as well as those generated by their descendant
nodes. Finally, compute the weight wi,j of each link (i, j) in
G′ from node weight wi, i.e., set wi,j = wi, where node j
is the parent of node i. Note that each link weight wi,j in
G′ represents the total number of times link (i, j) must be
activated in the superframe such that node i can forward wi
packets to its parent node j until all packets reach the root
node. Fig. 4a shows an example WSN G [34] while Fig. 4b
gives its mapping graph G′; the number next to a node and a
link shows its node and link weight, respectively. Note that
G′ can be constructed from G in polynomial time. FIGURE 7. A (4 × 3) grid. FIGURE 7. A (4 × 3) grid. FIGURE 7. A (4 × 3) grid. thus ρ = r. Recall that ρi is the number of slots for a node i
to accumulate energy to reach at least 1ϵ worth of energy,
starting from the time the node has Ai,t < 1ϵ. Note, all
three topologies are bipartite graphs [29]. Briefly, a graph is
bipartite if its nodes can be placed into two sets, namely Set-1
and Set-2, with links between nodes in Set-1 and Set-2 only. thus ρ = r. B. PROBLEM STATEMENT Note that a link can be scheduled only when each of its end
nodes has at least 1ϵ amount of energy. Let ti be the most
recent time node i transmits/receives a packet. |SE| = max
ρi
X
jϵN(i)
wi,j +
X
jϵN(i)
wj,i
(1) (1) where N(i) is the set of node i’s neighbours. The following theorem states that the LSHUS problem is
intractable. Consider Fig. 2a to illustrate the aforementioned notation. The battery at each node in Fig. 2a has energy level b1 = 3,
and b2 = b3 = b4 = 2, and harvesting time at each
node is r1 = 2, r2 = 6, r3 = 5, and r4 = 7 time slots. Assume ηi = 1 and µi = 0 for all batteries. At time t = 1,
the available energy of node 1 is A1,1 = 0.5ϵ, while at time
t = 2, it increases to A1,2 = 1ϵ. Thus, the earliest time
node 1 can transmit/receive is T1 = 2. For the other nodes,
Fig. 2a shows T2 = 6, T3 = 5, and T4 = 7. The earliest
time in which node 1 and 2 can transmit/receive is therefore
t1,2 = max(2, 6) = 6. The other two nodes have t3,1 = 5,
and t4,3 = 7. Notice that the first four slots in schedule S are
empty because the smallest ti,j is five. Assume a scheduler
selects link l1,2 first at time 6; thus t1 = t2 = 6, and the next Theorem 1: LSHUS is NP-complete. Theorem 1: LSHUS is NP-complete. Proof:
In [34], Ergen et al. address the following
NP-complete scheduling problem, referred to as ScheduleEV:
consider a WSN G(V, E) that forms a tree in which each
node i except the root node generates ωi > 0 packets. The
scheduling problem is to find a supeframe with the minimum
length such that all nodes can send their packets to the root
node. Note that ScheduleEV and LSHUS use the same con-
flict graph. The proof is by reduction from ScheduleEV to LSHUS. Specifically, an instance of WSN G(V, E) for ScheduleEV
can be mapped into an instance of WSN G′(V ′, E′) for 104726 VOLUME 7, 2019 VOLUME 7, 2019 T. Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries FIGURE 4. C. PROBLEM ANALYSIS This section analyzes the effects of harvesting time, bat-
tery storage efficiency, leakage and capacity on the sched-
ule length on some well-known graphs. They include Line
(Fig. 5), Binary Tree (Btree) (Fig. 6), and Grid (Fig. 7). Consider each node has equal harvesting time r > 0, battery
capacity b ≥1, and link weight w ≥1. Further, the storage
efficiency is set to η = 1 and the leakage rate is set to µ = 0; Consider each link has weight w = 1. For r = 1, schedule
links in the set {(2+4i, 3+4i), (5+4i, 4+4i)} in Slot-1, for
i = 0, 1, · · · , ⌊(n −2) / 4⌋; e.g., for n = 9, Slot-1 contains
links {(2, 3), (5, 4), (6, 7), (9, 8)}. Then, reverse the direction
of each link in Slot-1, and schedule the reversed link in Slot-2;
e.g., Slot-2 contains links {(3, 2), (4, 5), (7, 6), (8, 9)}. Next, 104727 VOLUME 7, 2019 T. Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries schedule links in the set {(1 + 4i, 2 + 4i), (4 + 4i, 3 +
4i)} in Slot-3, for i = 0, 1, · · · , ⌊(n −2) / 4⌋; e.g., links
{(1, 2), (4, 3), (5, 6), (8, 7)}, and finally reverse the links for
Slot-4; e.g., links {(2, 1), (3, 4), (6, 5), (7, 8)}. as follows. First, combine links of T1 scheduled in one slot
with the corresponding links of T2 that are scheduled in the
same slot number. Notice that no links in T1 interfere with
any links in T2, and vice versa. Then, insert each of the four
links incident at R into a slot that does not contain any link that
interferes with it. More specifically, link (1, 2), (2, 1), (1, 3),
and (3, 1) are inserted into a slot that contains link (8, 4),
(4, 8), (12, 24) and (24, 12) respectively. Using the same
steps, one can schedule links in a five-level tree using the six
slots schedules of its two four-level subtrees. Repeating the
process, one can always generate the link schedules of a tree
with k levels in six slots from the schedules of its two subtrees
with (k −1) levels, for any k ≥4. C. PROBLEM ANALYSIS For r > 1, use the same link schedule described above
for case r = 1 each time all nodes have sufficient energy. Thus, this case will have four non-empty slots. That is the
first r−1 slots are empty as all nodes have insufficient energy. Therefore, the first non-empty slot is slot r, which is followed
by r −1 empty slots. After that, the scheduler can activate
other links in slot 2r. Similarly, the third and fourth non-
empty slots are slot 3r and 4r, respectively. When each link has weight w ≥1, repeat the schedule
w times, and thus, the link schedule has 4wr slots, which is
optimal for a Line with n ≥3 for the following reason. For
n ≥3, some nodes in Line have four bidirectional links; each
of which can be activated no earlier than every r slots. Further,
each link has to be activated w times, and thus the schedule
requires 4wr slots. (
)
,
y
≥
For r > 1, the first r −1 slots contain no links because
each node requires harvesting time r. Then, use the same link
schedules described for case r = 1, except after scheduling
the first set of links at slot r, each node can transmit or receive
only after every r slots, i.e., there are r −1 empty slots in
between every non-empty slots. Thus, |S| = 6r for w = 1. When w ≥1, one can repeat the above schedule w times,
and thus the link schedule uses 6wr slots. The optimality
of the schedule is shown as follows. For L
≥3, some
nodes in BTree have six bidirectional links, each of which
can be activated no earlier than every r slots of harvesting
time. Since each link must be activated w times, the schedule
requires 6wr slots. For r > 1, the first r −1 slots contain no links because
each node requires harvesting time r. Then, use the same link
schedules described for case r = 1, except after scheduling
the first set of links at slot r, each node can transmit or receive
only after every r slots, i.e., there are r −1 empty slots in
between every non-empty slots. Thus, |S| = 6r for w = 1. When w ≥1, one can repeat the above schedule w times,
and thus the link schedule uses 6wr slots. 2) BINARY TREE Consider a Btree with L ≥2 levels, n nodes and bidirec-
tional links between nodes. Consider a complete binary tree,
i.e., n = 2L −1. Note that for 2L−1 ≤n < 2L −1, one
can include some dummy nodes to form a complete tree. The
root node is at level k = 1. Its two children are at level k = 2. Nodes are labelled consecutively from left to right level by
level, e.g., the root node is node 1, and the rightmost node at
the lowest level k = L is node 2k −1. Thus, level k contains
2k−1 nodes; the nodes are labeled 2k−1, 2k−1+1, · · · , 2k −1. As a bipartite graph, Set-1 contains nodes at odd levels of
Btree, and Set-2 has nodes at even levels. The secondary
interference in Btree can occur only between (i) pair of nodes
from different levels having a common neighbor, e.g., nodes 1
and 4 activating links (1, 2) and (4, 8), and (ii) pair of nodes
with the same parent, e.g., nodes 4 and 5 with links (4, 2)
and (5, 10). Consider a Btree with L ≥2 levels, n nodes and bidirec-
tional links between nodes. Consider a complete binary tree,
i.e., n = 2L −1. Note that for 2L−1 ≤n < 2L −1, one
can include some dummy nodes to form a complete tree. The
root node is at level k = 1. Its two children are at level k = 2. Nodes are labelled consecutively from left to right level by
level, e.g., the root node is node 1, and the rightmost node at
the lowest level k = L is node 2k −1. Thus, level k contains
2k−1 nodes; the nodes are labeled 2k−1, 2k−1+1, · · · , 2k −1. C. PROBLEM ANALYSIS The optimality
of the schedule is shown as follows. For L
≥3, some
nodes in BTree have six bidirectional links, each of which
can be activated no earlier than every r slots of harvesting
time. Since each link must be activated w times, the schedule
requires 6wr slots. 3) GRID Let (row×col) Grid be a bidirectional grid topology that
contains n = row × col nodes. Nodes are labelled starting
from number one sequentially from left-to-right in row-wise
manner. For example, for the (4 × 3) Grid in Fig. 7, row = 1
contains nodes {1, 2, 3}, and col = 3 has nodes {3, 6, 9, 12}. One can consider a (row × col) Grid is constructed from row
(col) Line graphs, each of which has col (row) nodes. Each
row Line graph is labelled as R1, R2, . . . , Rrow, and each col
Line graph as C1, C2, . . . , Ccol. The secondary interference
in Grid can occur only between pair of nodes from different
row or column that share a common neighbor. For example,
consider nodes 1 and 5 in Fig. 7 where node 2 is their common
neighbor. This means there is interference at node 2 when
links (1, 2) and (5, 8) are activated simultaneously in the same
slot. As a bipartite graph, Set-1 contains nodes at odd levels of
Btree, and Set-2 has nodes at even levels. The secondary
interference in Btree can occur only between (i) pair of nodes
from different levels having a common neighbor, e.g., nodes 1
and 4 activating links (1, 2) and (4, 8), and (ii) pair of nodes
with the same parent, e.g., nodes 4 and 5 with links (4, 2)
and (5, 10). A. PROPOSITIONS Algorithm LS-rWSN schedules links according to the earliest
time in which they have sufficient energy. It relies on the
following Proposition 4, 5 and 6. Let ti be the time in which
node i last draws energy from its battery, and bi,ti be the
energy level of the battery at node i at time ti. In case (ii), the battery cannot retain its stored energy. One
can see that when ˆµi = 0, Eq. (2) reduces to Ai,t = 1/ri. In case (iii), when ηi = 0, no harvested energy can be stored
in the battery. For this case, the harvesting time is set to
ˆri = ∞. Thus, the value of the aforementioned term (ii) in
Eq. (2) becomes zero. Cases (ii) and (iii) represent a possibly
faulty battery. Finally, case (iv) considers the effects of battery
storage efficiency and leakage on the available energy of each
node. Proposition 4 computes Ai,t, the amount of energy at node i
that can be used to transmit/receive packets at time t ≥1,
computed as Ai,t = bi,t + 1/ri. For brevity, in the following
propositions, we define ˆri = ri/ηi, and ˆµi = 1 −µi. Further,
we use τi to denote the time span between time ti and t,
i.e., τi = t −ti. This paper considers 0 ≤µi < 1 and 0 < ηi ≤1, i.e., only
case (i) and (iv). On the other hand, the work in [15] considers
only case (i), and thus our work generalizes the battery model
in [15] to also consider battery storage efficiency and leakage. Proposition 5 computes the number slots ρi = t−ti needed
such that Ai,t = 1ϵ. The condition Ai,ti < 1 means that the
amount of available energy at node i at time ti is insufficient
to transmit/receive one packet. Note that Ai,ti < 1 implies
ri > 1 and bi,t < 1 −1/ri. This paper considers 0 ≤µi < 1 and 0 < ηi ≤1, i.e., only
case (i) and (iv). On the other hand, the work in [15] considers
only case (i), and thus our work generalizes the battery model
in [15] to also consider battery storage efficiency and leakage. Proposition 2: The optimal link schedule for Btree with
L ≥3 levels has |S| = 6wr time slots. Proposition 2: The optimal link schedule for Btree with
L ≥3 levels has |S| = 6wr time slots. Proof: This proof first outlines the steps used to obtain
a link schedule with 6wr time slots for the following cases:
(i) r = 1 and (ii) r > 1. Then, it shows that the result is
optimal. For r = 1, links in Btree with three levels can be scheduled
in six slots: Slot-1 = {(1, 2), (3, 6)}, Slot-2 = {(2, 1), (6, 3)},
Slot-3 = {(1, 3), (2, 4)}, Slot-4 = {(3, 1), (4, 2)}, Slot-5 =
{(2, 5), (3, 7)}, and Slot-6 = {(5, 2), (7, 3)}. The link sched-
ules for a four-level Btree, see Fig. 6, can be generated from
schedules of two Btree with three levels as follows. Denote
the larger tree as T with root R, and the two smaller trees
as T1 and T2 with root R1 and R2, respectively. Tree T is
constructed from T1 and T2 by connecting R to R1 with
two bidirectional links, and R to R2 with two other links,
i.e., T1 and T2 are the left and right subtrees of T respectively. Thus, renumbering the nodes in T, the label of nodes R, R1,
and R2 are 1, 2, and 3 respectively. Links in T are scheduled Proposition 3: The optimal link schedule length for a Grid
of size row ≥3 and col ≥3 is |S| = 8wr. Proposition 3: The optimal link schedule length for a Grid
of size row ≥3 and col ≥3 is |S| = 8wr. Proof: From Proposition 1, the link schedule of each
Line graph requires 4wr slots. Next, the following describes
how to produce the schedule for Grid with length 8wr. Finally, the length is shown optimal. Consider the schedule of links in row Line graphs. One can
observe that links in odd rows can be activated in 4wr slots. Similarly, for links in even rows. Further, except for links in
two consecutive rows, there is no other interference. Without
loss of generality, consider links in R2 and R3 in Fig. 7. Note
that there is interference only between nodes in each (2 × 2)
Grid, e.g., nodes {4, 5} at R2 and {7, 8} at R3, and when 104728 VOLUME 7, 2019 VOLUME 7, 2019 T. Proposition 2: The optimal link schedule for Btree with
L ≥3 levels has |S| = 6wr time slots. Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries the energy harvested at time ti by slot ti; i.e., 1/ˆri = 0. Thus,
bi,ti+1 = (1−µi)bi,ti. Similarly, one can compute the amount
of energy available at the battery at time slot ti + 2 from the
stored energy at ti + 1, minus the amount of energy that has
leak between time ti + 1 to ti + 2, plus the stored energy
from the energy harvested in time slot ti + 1. Thus, the result
is bi,ti+2 = (1 −µi)bi,ti+1 + 1/ˆri. Substituting bi,ti+1 with
(1 −µi)bi,ti gets bi,ti+2 = (1 −µi)[(1 −µi)bi,ti] + 1/ˆri =
(1 −µi)2bi,ti + 1/ˆri. Next, for time ti + 3, this case obtains
bi,ti+3 = (1 −µi)3bi,ti + (1 −µi)/ˆri + 1/ˆri. Repeating
the step for time ti + 4, · · · , t −1, t, one can generate the
stored energy at the battery at the beginning of time t, i.e.,
bi,t = (1 −µi)t−tibi,ti + Pt−ti−2
k=0
(1 −µi)k/ˆri. Note that, for
the HUS model, the available energy at node i at time t is the
sum of its available energy at the beginning of time t and the
energy harvested at time t. In particular, the model has Ai,t =
bi,t +1/ri or Ai,t = (1−µi)τibi,ti +Pτi−2
k=0 (1−µi)k/ˆri +1/ri. However, bi,t is bounded by the battery capacity bi, which
implies Ai,t ≤bi + 1/ri. their links are in opposite directions, e.g., links (4, 5) and
(8, 7) that incur interference at nodes 5 and 7. To prevent
any interference, each slot must contain links of the same
directions, e.g., links (4, 5) and (7, 8). Thus, links in all rows
can be scheduled in 4wr slots. As an example, for Fig. 7 with
r = w = 1, the four slots are: Slot-1 = {(1, 2), (4, 5),
(7, 8), (10, 11)}, Slot-2 = {(2, 1), (5, 4), (8, 7), (11, 10)},
Slot-3 = {(2, 3), (4, 5), (8, 9), (11, 12)}, and Slot-4 = {(3, 2),
(6, 5), (9, 8), (12, 11)}. Use the same construction for links
in all columns. More specifically, these links are scheduled
in 4wr slots. Thus, all links in Grid can be scheduled in
4wr + 4wr = 8wr slots. IV. SOLUTION Section IV-A describes three propositions relied upon by the
proposed greedy algorithm, namely called Link Scheduler for
rechargeable WSN (LS-rWSN), to solve LSHUS. The details
of LS-rWSN is presented in Section IV-B. Ai,t = min(bi + 1/ri, Ai,ti + (τi −1)/ri)
(3) (3) Proposition 2: The optimal link schedule for Btree with
L ≥3 levels has |S| = 6wr time slots. The constructed schedules are
optimal because for row ≥3 and col ≥3, some nodes in
(row × col) Grid have eight bidirectional links, e.g., node 5
and 8 in Fig. 7. Since each node can be activated no earlier
than every r slots of harvesting time for w times, the link
schedule of Grid requires 8wr slots. Proposition 1, 2, and 3 show the effect of energy harvesting
time and link weight on the schedule length. These proposi-
tions indicate that, for r > 1 and w ≥1, there are 4w(r −1),
6w(r −1), and 8w(r −1) empty slots in the TDMA schedules
for Line, Btree, and Grid, respectively, and thus the schedule
lengths increase as much. Further, all propositions show that
the battery capacity of nodes and network size do not affect
the schedule length for the Line, Btree, and Grid. In addition,
the lower bound in Eq. (1) is tight for the fixed topologies
when µ = 0 and η = 1. Consider the following terms in Eq. (2): (i) ˆµτi
i bi,ti,
(ii) Pτi−1
k=0
ˆµk
i
ˆri , and (iii) 1/ri. Term (i) represents the amount
of energy stored in the battery of node i up to time ti after
accounting for energy leakage. Term (ii) shows the amount
of energy that can be added to the battery from time ti to the
start of time t. The last term shows the amount of harvested
energy, in the capacitor, at time t that is available for use by
node i. Consider four possible tuples (ηi, µi) in Eq. (2): (i) (ηi = 1,
µi = 0), (ii) (0 < ηi ≤1, µi = 1), (iii) (ηi = 0, 0 ≤
µi < 1), and (iv) (0 < ηi < 1, 0 < µi < 1). For case (i),
the battery has 100% storage efficiency and no leakage, and
hence, Eq. (2) reduces to the following equation: Proposition 5: Given Ai,ti < 1, the number of slots ρi =
t −ti before a node i has Ai,t = 1 is given as Case (i): ηi = 1 B. LS-rWSN The function
greedily schedules links closer to each other when there
is no interference. More specifically, function ORDER(K)
performs the following steps: (i) Select a link (i, j) ∈K;
(ii) Move link (i, j) from set K into a set K ′; (iii) Find a link
(m, k) ∈K that does not interfere with any link in K ′, where k
is node j’s neighbor. Note that there is no interference between
links (i, j) and (m, k); (iv) Move (m, k) from K to K ′; (v) Set
j = k and repeat (iii) until no such link can be found in K; (vi)
repeat step (i) until K is empty. Without loss of generality,
when there is more than one candidate link for selection in
Step (i) and (iii), select a link (u, v), where u is a node with
the smallest label, and v is node u’s neighbor with the smallest
label. Recall that Eq. (4) and Eq. (5) apply only when Ai,ti < 1,
which implies ri > 1 and bi,t < 1 −1/ri. For bi,t = 0,
Eq. (4) has ρi = ri, which is consistent with the fact that when
the battery is leakage free and has perfect storage efficiency,
it takes ri slots to accumulate 1ϵ of energy. In addition, Eq. (5)
requires ηi −bi,tiµiri > 0, or ηi
ri > µibi,ti. In other words,
the amount of harvested energy to be stored in the battery
must be larger than the energy leakage from the battery. Otherwise, the battery is always empty. Eq. (5) also requires
µi −µiri + ηi > 0, or
ηi
ri−1 > µi. Let T (bi,ti, ri, ηi, µi) be a
function to compute the value of ρi for Eq. (6). Specifically,
for (ηi = 1, µi = 0) and (ηi ̸= 1, µi ̸= 0), the function
produces Eq. (4) and Eq. (5), respectively. The following Proposition 6 computes the next value of
Ti after node i transmits/receives one packet at time ti. Let
αi,ti be a Boolean variable such that αi,ti = 1 (αi,ti = 0) if at
time ti node i has Ai,ti < 1ϵ (Ai,ti ≥1ϵ). Further, set σi = ti
(σi = ti + 1) when Ai,ti < 1ϵ (Ai,ti ≥1ϵ). Line 11 initializes t with the earliest slot. Lines 12-26
repeatedly schedule each link li,j ∈K ′ in order. B. LS-rWSN This section provides the details of the new algorithm,
i.e., LS-rWSN. The algorithm selects each non-interfering
link with end nodes that have sufficient energy to trans-
mit/receive one packet at the earliest time. It uses the conflict
graph CG to check for interfering links. ˆµρi
i bi,ti +
ρi−2
X
k=0
ˆµk
i
ˆri
+ 1/ri ≥1
(6) (6) For case (i), setting ˆµi = 1 and ˆri = ri in Eq. (6) obtains
bi,ti + ρi/ri ≥1. Since Ai,ti = bi,ti + 1/ri, case (i) produces
Ai,ti −1/ri + ρi/ri ≥1. Thus, ρi = ⌈ri(1 −Ai,ti + 1)⌉as
shown in Eq. (4). In case (ii), ˆµi ̸= 1, and thus one can use
the geometric series to produce Pρi−1
k=0
ˆµk
i
ˆri = 1
ˆri
ˆµρi
i −1
ˆµi−1 . Thus,
Eq. (6) becomes ˆµρi
i bi,ti + 1
ˆri ( ˆµρi
i −1
ˆµi−1 −ˆµρi−1) + 1/ri ≥1,
which is used to obtain ρi in Eq. (5). For case (i), setting ˆµi = 1 and ˆri = ri in Eq. (6) obtains
bi,ti + ρi/ri ≥1. Since Ai,ti = bi,ti + 1/ri, case (i) produces
Ai,ti −1/ri + ρi/ri ≥1. Thus, ρi = ⌈ri(1 −Ai,ti + 1)⌉as
shown in Eq. (4). In case (ii), ˆµi ̸= 1, and thus one can use
the geometric series to produce Pρi−1
k=0
ˆµk
i
ˆri = 1
ˆri
ˆµρi
i −1
ˆµi−1 . Thus,
Eq. (6) becomes ˆµρi
i bi,ti + 1
ˆri ( ˆµρi
i −1
ˆµi−1 −ˆµρi−1) + 1/ri ≥1,
which is used to obtain ρi in Eq. (5). g p
G
g
LS-rWSN first initializes ti to the last time slot node i
draws energy from its battery, and Ai,ti to the amount of
energy that node i can use at time ti to zero; see Lines 1-5. It also sets Ti to ρi slots, i.e., the solution for Eq. (6). Recall
that Ti is the earliest time node i has 1ϵ energy. LS-rWSN
starts at t = 0 and the battery is initially empty. Lines 6-8
compute the earliest time link (i, j) can be scheduled, while
Lines 9 generate a set K to record link (i, j) that has the
earliest activation time. Line 10 uses function ORDER(K) to
sort links in K. It aims to maximize the number of links that
can be scheduled at time t without interference. A. PROPOSITIONS Proposition 4: The amount of energy (in unit of ϵ) that
node i can use at time slot t > ti is, Ai,t = min(bi + 1/ri, ˆµτi
i bi,ti +
τi−2
X
k=0
ˆµk
i
ˆri
+ 1/ri)
(2) Proposition 5 computes the number slots ρi = t−ti needed
such that Ai,t = 1ϵ. The condition Ai,ti < 1 means that the
amount of available energy at node i at time ti is insufficient
to transmit/receive one packet. Note that Ai,ti < 1 implies
ri > 1 and bi,t < 1 −1/ri. (2) Proof: The stored energy at time ti + 1, i.e., bi,ti+1,
is computed by subtracting the energy due to leakage from
the battery, i.e., µibi,ti and adding energy harvested at slot ti,
i.e., 1/ˆri. However, for HUS with ri > 1, node i spends all Proposition 5: Given Ai,ti < 1, the number of slots ρi =
t −ti before a node i has Ai,t = 1 is given as Case (i): ηi = 1 104729 104729 VOLUME 7, 2019 T. Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries and µi = 0 and µi = 0 and µi = 0 packet at time ti. However, the primary interference does
not allow a node to transmit/receive more than one packet
at the same time slot. Thus, the earliest time the node can
transmit/receive a packet is in the next slot, i.e., Ti = ti + 1. For this case, in Eq. (7), αi,ti = 0 and σi = ti + 1. ρi = ⌈ri(1 −Ai,ti) + 1⌉
(4)
Case (ii): 0 < ηi < 1 and 0 < µi < 1
ρi = ⌈
log(
µi−µiri+ηi
ηi−bi,tiµiri(1−µi))
log(1 −µi)
+ 1⌉
(5) (4) i
The work in [15] considers battery with 100% storage
efficiency and zero leakage, i.e, ηi = 1 and µi = 0. For this
case, using Eq. (4), Eq. (7) becomes Ti = σi+αi,t(ri(1−Ai,t)),
as given in [15]. Further, this case produces Ti = ri for
bi,t = 0 or t = 0 because each battery is initially empty. (5) Proof: Node i needs at least (1 −Ai,ti)ϵ extra energy to
reach 1ϵ unit of energy. Eq. (2) is used to compute the time
span ρi = t−ti such that Ai,t ≥1. More specifically, compute
the smallest ρi that satisfies the following: B. LS-rWSN Each selected
link in Line 14 does not cause interference or is interfered
by links that have been scheduled in slot t; see the condi-
tion in Line 13. Each slot in S is initially empty. Note that
function CONFLICT() uses a matrix M of size |E|2 that
contains Boolean variables to represent the conflict graph of
the network; i.e., M[a, b] is set to ‘‘1’’ if there is interfer-
ence between links a and b. Line 15 decrements the weight
of each selected link li,j by one. Once the weight reaches
zero, Line 17 removes the link from contention. Further,
Lines 19-20 use function COMPUTE_Ai,t() that implements
Eq. (2) to recompute the available energy Ai,t and Aj,t at
each end node of the selected link. Line 21 then sets the last
time that the end nodes of the selected link use energy to the Proposition 6: The next earliest time slot when node i has
sufficient energy to transmit/receive one packet is Proposition 6: The next earliest time slot when node i has
sufficient energy to transmit/receive one packet is Ti = σi + αi,tiρi
(7) (7) Proof: The next value of Ti depends on the remaining
available energy in node i, i.e., Ai,ti, harvesting time ri, and
the battery’s storing efficiency ηi and leakage rate µi. Eq. (7)
considers two cases. First, when Ai,ti < 1ϵ, node i needs
(1 −Ai,ti)ϵ extra energy to transmit/receive the next packet. Proposition 5, i.e., Eq. (4) and (5), is used to compute the
number of slots ρi for node i to accumulate the extra energy. Thus Ti = ti+ρi, and in Eq. (7), αi,ti = 1, and σi = ti. Second,
when Ai,ti ≥1ϵ, after node i uses 1ϵ of energy at time ti,
the node still has sufficient energy to transmit/receive another VOLUME 7, 2019 104730 104730 VOLUME 7, 2019 T. Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries TABLE 3. Parameter values used in the evaluation. TABLE 3. Parameter values used in the evaluation. TABLE 3. Parameter values used in the evaluation. current time, and COMPUTE_Ti() in Lines 22-23 use Eq. (7)
to recompute the Ti and Tj of the two end nodes. Finally,
function UPDATE_tcd() in Line 24 recomputes the earliest
time schedule of each link that has vi or vj as one of its end
nodes. B. LS-rWSN Algorithm 1 LS-rWSN: A Greedy Algorithm That Schedules
Links According to the Earliest Activation Time
Input: G(V, E), ri, bi, µi, ηi of each node i ∈V, weight wi,j
of each link li,j ∈E, and conflict graph CG
Output: Superframe S
1: for each node i ∈V do
2:
ti ←0
3:
Ai,ti ←0
4:
Ti ←T (0, ri, ηi, µi)
5: end for
6: for each link li,j ∈E do
7:
ti,j ←max(Ti, Tj)
8: end for
9: K = {node li,j in CG with min{ti,j}}
10: K ′ = ORDER(K)
11: t ←min{ti,j}
12: for each li,j ∈K ′ do
13:
if NOT CONFLICT(li,j, S[t]) then
14:
S[t] ←S[t] ∪li,j
15:
wi,j ←wi,j −1
16:
if wi,j = 0 then
17:
remove node li,j from CG
18:
end if
19:
Ai,t ←COMPUTE_Aα,t(t, i, µi, ηi)
20:
Aj,t ←COMPUTE_Aα,t(t, j, µj, ηj)
21:
ti ←tj ←t
22:
Ti ←COMPUTE_Tα(t, i, µi, ηi)
23:
Tj ←COMPUTE_Tα(t, j, µj, ηj)
24:
UPDATE_tcd(Ti, Tj)
25:
end if
26: end for
27: repeat Line 9-26 until all wi,j = 0 Algorithm 1 LS-rWSN: A Greedy Algorithm That Schedules
Links According to the Earliest Activation Time Algorithm 1 LS-rWSN: A Greedy Algorithm That Schedules
Links According to the Earliest Activation Time Algorithm 1 LS-rWSN: A Greedy Algorithm That Schedules
Links According to the Earliest Activation Time
Input: G(V, E), ri, bi, µi, ηi of each node i ∈V, weight wi,j
of each link li,j ∈E, and conflict graph CG
Output: Superframe S
1: for each node i ∈V do
2:
ti ←0
3:
Ai,ti ←0
4:
Ti ←T (0, ri, ηi, µi)
5: end for
6: for each link li,j ∈E do
7:
ti,j ←max(Ti, Tj)
8: end for
9: K = {node li,j in CG with min{ti,j}}
10: K ′ = ORDER(K)
11: t ←min{ti,j}
12: for each li,j ∈K ′ do
13:
if NOT CONFLICT(li,j, S[t]) then
14:
S[t] ←S[t] ∪li,j
15:
wi,j ←wi,j −1
16:
if wi,j = 0 then
17:
remove node li,j from CG
18:
end if
19:
Ai,t ←COMPUTE_Aα,t(t, i, µi, ηi)
20:
Aj,t ←COMPUTE_Aα,t(t, j, µj, ηj)
21:
ti ←tj ←t
22:
Ti ←COMPUTE_Tα(t, i, µi, ηi)
23:
Tj ←COMPUTE_Tα(t, j, µj, ηj)
24:
UPDATE_tcd(Ti, Tj)
25:
end if
26: end for
27: repeat Line 9-26 until all wi,j = 0 Algorithm 1 LS-rWSN: A Greedy Algorithm That Schedules
Links According to the Earliest Activation Time links have wi,j = 0. V. EVALUATION V. EVALUATION LS-rWSN has been implemented in C++. All experiments
are conducted on a computer with an Intel Core i7 CPU @
3.4 GHz and 16 GB of memory. Section V-A analyzes the
effect of energy harvesting time, battery leakage, and storage
efficiency on the superframe length |S|. Then, Section V-B
shows the effect of battery capacity on |S|. Section V-C
contains an evaluation of LS-rWSN versus the theoretical
bounds of |S|. Finally, Section V-D discusses the running
time of LS-rWSN. Our evaluation encompasses two types of
networks: (i) fixed topologies, namely, Line, BTree, and Grid,
with 20 to 100 nodes, and (ii) arbitrary networks with 20 to
50 nodes deployed uniformly deployed on a 40 × 40 m2 area. Nodes have a transmit and an interference range of 15 and
30 meters, respectively. Each presented result is an average
over 100 random node deployments. Table 3 lists the param-
eter values used in our evaluation. For an example, consider the rWSN and conflict graph CG
shown in Fig. 2. Lines 1-5 of LS-rWSN set t1 = t2 =
t3 = t4 = 0, A1,t1
= A2,t2
= A3,t3
= A4,t4
= 0,
T1 = T (0, r1, η1, µ1) = T (0, 2, 1, 0) = 2, T2 = 6, T3 = 5,
and T4 = 7. Lines 6-8 compute t1,2 = max (2, 6) = 6,
t3,1 = 5, and t4,3 = 7. Line 9 places link l3,1 into the
set K, and thus Line 10 obtains K ′ = l3,1, and Line 11 sets
t = 5. Line 13 finds that l3,1 has no conflict with other
links, and thus Line 14 inserts the link into S[5], and Line 15
reduces w3,1 by one and hence it becomes zero. Lines 19-20
compute A3,5 = 0.2 and A1,5 = 2, while Line 21 sets
t3 = t1 = t = 5. Lines 22-23 then obtain T3 = 11 and
T1 = 6. Line 24 updates the earliest time that links can be
scheduled, i.e., t1,2 = max (6, 6) = 6, t3,1 = 11, and
t4,3 = 7. Line 27 repeats the steps from Line 9 until all B. LS-rWSN Fig. 3b shows the schedule generated by
LS-rWSN, i.e., S = [ S[5] = {l3,1}, S[6] = {l1,2}, S[10] =
{l4,3}, S[12] = {l1,2}, S[17] = {l4,3}, S[18] = {l1,2}]. The generated schedule contains 12 empty slots as nodes are
unable to transmit/receive due to insufficient energy. gy
Proposition 7: The
time
complexity
of
LS-rWSN
is
O(W|E|2), where W =
P (wi,j). P
(i,j)∈|E|
(
i
O ( j) | |
Proof: Lines 1-5 require O(|V|), while Lines 6-8 require
O(|E|). Line 9 inserts at most |E| links and thus take
O(|E| log |E|). Line 10 requires at most O(|E|2) to sort links
in K using the function ORDER(K). Line 11 takes O(1). Line 13 uses a matrix M to represent the conflict graph CG. it
takes O(|E|2) to build the CG and M. Note that LS-rWSN
constructs CG and M only once. The function in Line 13
needs to access the matrix |S[t]| times or O(|E|) to check
for interference between a selected link (i, j) and the already
scheduled links in S[t]. Lines 14 to 23 take O(1) each. Line 24
requires O(|V|). Thus, the for loop in Lines 12-26 takes at
most O(|E|2) since |V| ≤|E|. Finally, Line 27 repeats
Lines 9-26 W times, and hence the time complexity of LS-
rWSN is O(W|E|2). B. LS-rWSN The steps from Line 9 is repeated until all links have
wi,j = 0. 3) EFFECT OF STORAGE EFFICIENCY To study the effect of storage efficiency ηi, in this section,
the leakage rate µi is set to zero. Fig. 9 shows the resulting
schedule length |S| for ri = 1, 5, 10, 15, 20 and five values
of storage efficiency ηi. Similar to Fig. 8, from left to right,
each bar for each harvesting time is the result for network
size 20, 30, 40, or 50, respectively. Further, each bar shows
the average value of |S| when ηi is gradually decreased from
1 to 0.6. As shown in Fig. 9, when nodes require one slot to
harvest energy, i.e., ri = 1, decreasing ηi from one to 0.6
has no effect on the superframe length |S|, i.e., |S| remains
at 196, 334, 591 and 820 for networks with 20, 30, 40 and
50 nodes, respectively. The reason is because when nodes
use the HUS model, and ri = 1, each node has 1ϵ worth
of harvested energy in each slot that can be directly used
irrespective of the energy level of its battery. However, for
ri > 1, decreasing ηi has a significant negative effect on |S|. For example, when ri = 15 and ηi drops from one to 0.6,
the superframe length |S| increases by 62.21% (from 979 to
1588 slots) when there are 20 nodes, 62.17% (1372 to 2225)
for 30 nodes, 62.77% (1719 to 2798) when there are 40 nodes
and 61.72% (2223 to 3595) for 50 nodes. Fig. 9 also shows
that decreasing storage efficiency consistently creates longer
superframe lengths |S|. Specifically, for ri = 20, decreasing
ηi from 0.9 to 0.8 increases the link schedule by 11.92%
(from 1443 to 1615 slots) or 20 nodes, 11.8% (from 2026
to 2265 slots) for 30 nodes, 12.28% (from 2532 to 2843
slots) for 40 nodes, and 11.62% (from 3278 to 3659) slots
for 50 nodes. The superframe length |S| increases because,
when ri > 1, nodes need time to harvest energy before they
have Ai,t = 1ϵ worth of energy to transmit. However, due to
storage efficiency, nodes take longer time to reach the said
minimum amount of energy to transmit or receive. time to accumulate sufficient energy to transmit or receive
a packet. time to accumulate sufficient energy to transmit or receive
a packet. A. EFFECT OF ri, µi AND ηi ON |S| The following experiments consider three factors: (i) har-
vesting time, (ii) battery leakage, and (iii) storage efficiency, 104731 VOLUME 7, 2019 VOLUME 7, 2019 T. Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries FIGURE 8. The effect of harvesting time and leakage rate on |S|. with
the
following
corresponding
values:
(i)
ri
=
1, 5, 10, 15, 20, (ii) µi
= 0.0, 0.01, 0.02, 0.03, 0.04, and
(iii) ηi = 0.6, 0.7, 0.8, 0.9, 1.0. Further, the battery capacity
is bi = 3. Each link has weight wi,j = 3. These experiments
use the HUS battery recharging model. Note that the prior
work in [15] has shown that the HUS model produces shorter
superframe lengths as compared to using the HSU model. 1) EFFECT OF HARVESTING TIME Fig. 8 shows the resulting superframe length |S| for various
energy harvesting time ri and leakage value µi when the
storage efficiency is ηi = 1. Specifically, from left to right,
each bar for each harvesting time is the result for network of
size 20, 30, 40, or 50, respectively. Further, each bar shows the
average |S| value when µi is gradually increased from zero
to 0.04. For example, the left (right) most bar for ri = 15
shows five results of |S| for network with 20 (50) nodes for
each of the five values of µi. Recall that µi = 0 means the
battery has no leakage. FIGURE 8. The effect of harvesting time and leakage rate on |S|. As shown in in Fig. 8, for µi = 0, increasing the energy
harvesting time of nodes significantly affects the schedule
length. The figure shows that for ri = 1 and |V| = 20,
the superframe length is |S| = 196. However, as the har-
vesting time increases, the length increases steadily, reaching
|S| = 1305 for ri = 20; this is an increase of 565.82%. This is because more nodes need a longer time to harvest
energy. Recall that ri = 1 means each node, which uses the
HUS mode, always has sufficient energy to transmit/receive
one packet in any slot. This means there is no constraint
on harvesting time. The negative effects of having larger ri
values are similar for each of the tested network sizes; namely,
|V| = 30, 40, 50. 2) EFFECT OF LEAKAGE RATE Fig. 8 shows when the energy harvesting time of nodes is
ri = 1, increasing µi from zero to 0.04 has no effect on the
schedule length. That is, |S| remains at 196, 334, 591 and
820 for networks with 20, 30, 40 and 50 nodes, respectively. However, with a longer harvesting time ri > 1, e.g., ri =
15, increasing µi from 0 to 0.04 significantly affects the
superframe length |S|. In particular, we see that |S| increases
from 2223 to 3187 slots for |V| = 50, an increase of 43.36%. The results are consistent for other tested network sizes,
i.e., an increase of 44.54% (from 979 to 1415 slots), 44.24%
(1372 to 1979), and 43.92% (1719 to 2474) for networks with
20, 30, and 40 nodes respectively. Fig. 8 also shows that in
general increasing the leakage rate creates longer superframe
lengths |S|. For example, for an energy harvesting time of
ri = 20, the link schedule increases by 16.6% (from 1639 to
1911 slots) for 20 nodes, 16.71% (from 2298 to 2682 slots)
for 30 nodes, 16.76% (from 2876 to 3358 slots) for 40 nodes,
and 16.47% (from 3703 to 4313 slots) for 50 nodes when we
increase the value of the leakage rate µi from 0.02 to 0.03. The reason the superframe length increases is because when
ri > 1, a node i needs to first accumulate energy until Ai,t =
1ϵ. However, due to storage leakage, it requires a longer 104732 104732 104732 VOLUME 7, 2019 T. Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries FIGURE 9. The effect of harvesting time and storage efficiency on |S|. 63.21% (2963 to 4836) for ri = 5, 10, 15, 20, respectively. The superframe length is longer because as nodes have an
imperfect battery, they require a longer time to accumulate
energy. Fig. 10 also shows that the increase in |S| is more
noticeable in denser networks; see the results for 100 nodes. One can observe that leakage rate and storage efficiency
together cause more severe negative effects on the super-
frame length |S| as compared to the effect of leakage rate
(Fig. 8) or storage efficiency (Fig. 9) separately. As an exam-
ple, consider the result for a network with 50 nodes and
ri = 20. For µi = 0.01 and ηi = 1, as per Fig. B. EFFECT OF bi, µi AND ηi ON |S| This section considers rWSN with 40 nodes and set wi,j = 3,
ri = 5, and battery capacity bi = 1, 5, 10, 15, 20. The
following sections discuss the effect of 1) battery capacity
only, 2) battery capacity and leakage rate, and 3) battery
capacity and storage efficiency. 4) EFFECT OF LEAKAGE RATE AND STORAGE EFFICIENCY 4) EFFECT OF LEAKAGE RATE AND STORAGE EFFICIENCY
This section aims to investigate the effect of battery leakage
rate together with storage efficiency on the schedule length. It compares the schedule length for two cases: (i) µi = 0
and ηi = 1, and (ii) µi = 0.01 and ηi = 0.7. Note that
the schedule length in case (i) is affected only by the energy
harvesting time of nodes and interference. The results for case
(i) are used to benchmark against the results for case (ii). This section aims to investigate the effect of battery leakage
rate together with storage efficiency on the schedule length. It compares the schedule length for two cases: (i) µi = 0
and ηi = 1, and (ii) µi = 0.01 and ηi = 0.7. Note that
the schedule length in case (i) is affected only by the energy
harvesting time of nodes and interference. The results for case
(i) are used to benchmark against the results for case (ii). This section aims to investigate the effect of battery leakage
rate together with storage efficiency on the schedule length. It compares the schedule length for two cases: (i) µi = 0
and ηi = 1, and (ii) µi = 0.01 and ηi = 0.7. Note that
the schedule length in case (i) is affected only by the energy
harvesting time of nodes and interference. The results for case
(i) are used to benchmark against the results for case (ii). As shown in Fig. 10, imperfect battery for case (ii) has
a significant effect on the schedule length |S| for all net-
work sizes. More specifically, for a rWSN with 20 nodes,
the superframe |S| increases by 37.61% (from 327 in case
(i) to 450 slots in case (ii)), 47.78% (653 to 965), 55.46%
(979 to 1522), and 63.68% (1305 to 2136), for ri = 5, 10,
15, 20, respectively. A similar negative effect is also noticed
for larger sized rWSNs. For example, when there are 50
nodes, the superframe length increases by 10.46% (975 to
1077), 47.44% (1482 to 2185), 55.24% (2223 to 3451), and 2) EFFECT OF LEAKAGE RATE 8, we find
that the superframe length is |S| = 3258, while from Fig. 9
it is |S| = 4166 for ηi = 0.7 and µi = 0. On the other hand,
as shown in Fig. 10, setting ηi = 0.7 together with µi = 0.01
results in |S| = 4836, which increases the length by 48.43%
(from 3258 to 4836) and 16.08% (4166 to 4836) as compared
to when considering only leakage rate and storage efficiency
only, respectively. FIGURE 9. The effect of harvesting time and storage efficiency on |S|. FIGURE 10. Effect of network sizes with varying ri on |S|. 1) EFFECT OF BATTERY CAPACITY To see the effect of battery capacity on the schedule length,
we set µi = 0 and ηi = 1. As shown in Fig. 11, µi = 0,
increasing bi from 1 to 20 only slightly reduces the super-
frame length |S|; there is only a 6.5% decrease (from 769 to
719 slots). As reported in [15], the battery capacity constraint
has negligible effect on the schedule length. 2) EFFECT OF LEAKAGE RATE FIGURE 10. Effect of network sizes with varying ri on |S|. To analyze the effect of battery capacity and leakage rate
µi on |S|, we set the storage efficiency to ηi = 1. From
Fig. 11, we see that increasing the battery capacity for µi =
0, 0.01, 0.02, 0.03, 0.04 has an insignificant effect on the
superframe length |S|. As an example, when µi = 0.01,
increasing bi from one to 20 reduces the superframe length
by 6.61%, i.e., from 772 to 721 slots. Similarly, there is only
a decrease of 4.26% (775 to 742 slots), 3.59% (779 to 751
slots), and 4.57% (787 to 751 slots) for µi value of 0.02, 0.03,
0.04, respectively, when bi increases from one to 20. Note that
the standard deviation values of |S| range between 117 to 131. 3) EFFECT OF STORAGE EFFICIENCY To evaluate the effect of battery capacity and storage effi-
ciency, consider the leakage rate µi = 0. As shown in Fig. 12,
increasing the battery capacity for ηi = 1, 0.9, 0.8, 0.7, 0.6
does not affect the superframe length significantly. For exam-
ple, when ηi = 0.9, increasing bi from 1 to 20 only reduces
the superframe length |S| from 792 to 721 slots. This is a
decrease of only 8.96%. Note that the standard deviation
values of |S| range between 84 to 126. 104733 VOLUME 7, 2019 VOLUME 7, 2019 T. Tony et al.: Link Scheduling in rWSNs With Imperfect Batteries FIGURE 11. The effect of battery capacity and leakage rate in a rWSN with
40 nodes. FIGURE 13. The performance of LS-rWSN in fixed topologies with various
number of nodes for µi = 0 and ηi = 1. FIGURE 13. The performance of LS-rWSN in fixed topologies with various
number of nodes for µi = 0 and ηi = 1. FIGURE 13. The performance of LS-rWSN in fixed topologies with various
number of nodes for µi = 0 and ηi = 1. FIGURE 11. The effect of battery capacity and leakage rate in a rWSN with
40 nodes. FIGURE 13. The performance of LS-rWSN in fixed topologies with various
number of nodes for µi = 0 and ηi = 1. FIGURE 12. The effect of battery capacity and storage efficiency in a
network with 40 nodes. ratio between its generated |S| and the lower bound |SE| in
Eq. (1). For both types of networks, we set wi,j = 3, bi = 3,
and ri = 1, 5, 10, 15, 20. 1) FIXED TOPOLOGIES This experiment considers rWSNs with nodes from 20 to
100, with an increment of 10 for the Line, BTree and Grid
topology. Note that a BTree with n nodes has L = ⌈log2 n⌉+1
levels, e.g., for n = 30, the resulting BTree has L = 5 levels. Thus, in this experiment, each BTree is not a complete binary
tree; its lowest level is not fully populated. For Grid, this
experiment sets row ≥col such that (row −col) is minimum,
e.g., for n = 20 and n = 80, we have (5×4) and (10×8) Grid,
respectively. As shown in Fig. 13, LS-rWSN always produces
the optimal superframe for Line for any values of ri. On the
other hand, for ri = 1, BTree and Grid produce R1 = 1.28
and R1 = 1.42, respectively. However, LS-rWSN has better
performance for larger values of ri. Specifically, when ri ≥5,
LS-rWSN has a performance ratio R1 of 1.04 and 1.02 for
BTree and Grid, meaning the superframe length is only 4%
and 2% away from the optimal value, respectively, for rWSNs
with 20 to 100 nodes. FIGURE 12. The effect of battery capacity and storage efficiency in a
network with 40 nodes. The results shown in Fig. 11 and Fig. 12 indicate that
increasing the capacity of imperfect batteries can only
slightly reduce schedule length. The results are consistent to
those reported in [15] where nodes have a battery with perfect
storage, i.e., µi = 0 and ηi = 1. 2) ARBITRARY NETWORKS For each network, consider µ = 0.01, and η = 0.7. The
average ratio R2 in Fig. 14 is obtained by averaging from
100 random node deployments. As shown in Fig. 14, for a
rWSN with 20 nodes and ri = 1, i.e., when nodes always have
energy, LS-rWSN achieves an average performance ratio R2
of 3.01. However, when nodes have a lower energy harvesting
constraint, i.e., r > 1 the performance of LS-rWSN improves
significantly. Specifically, for ri ≥5 and |V| = 20, LS-rWSN
on average produces superframes with R2 = 1.07, i.e., only
7% longer than the lower bound computed using Eq. (1). Fig. 14 also shows that except for ri = 1, increasing the
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Th
f
f LS WSN i
bit
t
k f 152 links), 273 (from 236 to 312), 470 (from 420 to 556),
758 (from 670 to 854) links when there are 20, 30, 40, and
50 nodes, respectively. The standard deviation of the running
time in Fig. 15 ranges between 6.35 and 17.66. VI. CONCLUSION This paper has considered link scheduling in rWSNs. It con-
siders the following factors: (i) energy harvesting time of
nodes, (ii) battery capacity, (iii) imperfect battery storage,
i.e., leakage and storage inefficiency, and (iv) activation fre-
quencies wi. It has presented a novel problem called LSHUS
that aims to produce a link schedule S with the minimum
length subject using the HUS battery recharging model. This
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time, battery leakage, and storage efficiency significantly
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an insignificant factor, increasing the schedule length no
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Jan. 2010. 104736 VOLUME 7, 2019 VOLUME 7, 2019
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Is the flood occurrence rate decreasing in Southeast Europe?
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Igor Leščešen
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English
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EGU23-6031, updated on 30 Sep 2023
https://doi.org/10.5194/egusphere-egu23-6031
EGU General Assembly 2023
© Author(s) 2023. This work is distributed under
the Creative Commons Attribution 4.0 License. Is the flood occurrence rate decreasing in Southeast Europe? Igor Leščešen1, Biljana Basarin1, Manfred Mudelsee2, and Robert L. Wilby3
1Department of Geography, Tourism and Hotel Management, Faculty of Sciences, University of Novi Sad, Trg Dositeja
Obradovića 3, Novi Sad, Serbia, email: igorlescsen@yahoo.com; biljana.basarin@dgt.uns.ac.rs
2Climate Risk Analysis, Kreuzstrasse 27, 37581 Bad Gandersheim, Germany, email: mudelsee@climate-risk-analysis.com
3Department of Geography, Loughborough University, Loughborough LE11 3TU, UK, email: r.l.wilby@lboro.ac.uk Floods are natural phenomena, which can turn into disasters and cause widespread damage,
health problems and deaths. This is particularly the case where rivers have been denied from their
natural floodplains and are limited by embankments, and where housing and industrial buildings
have been constructed in areas that are naturally liable to flooding. However, during the last few
decades, flood observations from different parts of Europe do not show a clear increase in flood
occurrence rate (Blöschl et al., 2019). In the present paper we present longer-term records of
winter and summer floods in one of the largest river of Southeast Europe, the Sava River for the
1926-2021 period. We analysed three group of events that were based on three flood protection
levels in Republic of Serbia defined for Sremska Mitrovica station. Regular protection level is set at
4120 m3/s and emergency flood defense level which is set at 5120 m3/s, that is, minor events are
up to the regular protection level, strong events are up to the emergency level and extreme events
are above emergency flood defense level. For the past 95 years, we find a decrease in both
summer and winter flood occurrence rates. The reduction in winter flood occurrence can partly be
attributed to reduced amount of precipitation during this period of the year. Further, on the basis
of these data and methods, we find that for the Sava River can be stated the following: (1)
downward but not significant trends in winter flood risk during the observed period, (2) Downward
trends of summer floods was also observed, with only strong events being statistically significant. This decrease can be partially due to a projected decrease in cyclone frequency in the
Mediterranean region. Presented results clearly demonstrate decreasing flood occurrence rate of
the Sava River, as a consequence of decreasing precipitation and increasing evaporation (due to
increasing temperature). References Blöschl, G., Hall, J., Parajka, J., Perdigão, A. P. R., Merz, B., Arheimer, B., Aronica, T. G., Bilibashi, A.,
Bonacci, O., Borga, M., Čanjevac, I., Castellarin, A., Chirico, B. G., Claps, P., Fiala, K., Frolova, N.,
Gorbachova, L., Gül, A., Hannaford, J., Harrigan, S., Kireeva, M., Kiss, A., Kjeldsen, R. T., Kohnová, S.,
Koskela, J. J., Ledvinka, O., Macdonald, N., Mavrova-Guirguinova, M., Mediero, L., Merz, R., Molnar,
P., Montanari, A., Murphy, C., Osuch, M., Ovcharuk, V., Radevski, I., Rogger, M., Salinas, L. J.,
Sauquet, E., Šraj, M., Szolgay, J., Viglione, A., Volpi, E., Wilson, D., Zaimi, K. & Živković, N. (2019). Changing climate both increases and decreases European river floods. Nature 573 (7772), 108–111. https://doi.org/10.1038/s41586-019-1495-6. Acknowledgements This research was supported by ExtremeClimTwin project, which has received funding from the
European Union’s Horizon 2020 research and innovation programme under grant agreement No
952384.
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Stem Cells Dev. 2011 Dec; 20(12):2053-64. 162 Wiese C, Rolletschek A, Kania G, Blyszczuk P, Tarasov KV, Tarasova Y, Wersto RP, Boheler KR, Wobus AM. Nestin expression-a property of multi-lineage progenitor cells? Cell Mol Life Sci. 2004; 61: 2510–2522. Willard-Mack CL. Normal structure, function, and histology of lymph nodes. Toxicol Pathol. 2006; 34(5):409-24. Wood SJ, Slater CR. The contribution of postsynaptic folds to the safety factor for neuromuscular transmission in rat fast- and slow-twitch muscles. J. Physiol. 1997 ; 500:165–76. Wu J, Yan LJ. Streptozotocin-induced type 1 diabetes in rodents as a model for studying mitochondrial mechanisms of diabetic β cell glucotoxicity. Diabetes Metab Syndr Obes. 2015 Apr 2; 8:181-8. Wulf GG, Luo KL, Jackson KA, Brenner MK, Goodell MA. Cells of the hepatic side population contribute to liver regeneration and can be replenished with bone marrow stem cells. Haematologica 2003; 88:368 –378. X Xin L, Lawson DA, Witte ON. The Sca-1 cell surface marker enriches for a prostateregenerating cell subpopulation that can initiate prostate tumorigenesis. Proc Natl Acad Sci USA 2005; 102:6942– 6947. Y Yaffe D. Retention of differentiation potentialities during prolonged cultivation of myogenic cells. Proc Natl Acad Sci U S A. 1968 Oct; 61(2):477-83. Yanagisawa M. Stem cell glycolipids. Neurochem Res. 2011 Sep; 36(9):1623-35. Yang H, Mujtaba T, Venkatraman G, Wu YY, Rao MS, Luskin MB. Region-specific differentiation of neural tube-derived neuronal restricted progenitor cells after heterotopic transplantation. Proc Natl Acad Sci U S A. 2000 Nov 21;97(24):13366-71. 163 Yee WC, Pestronk A, Alderson K, Yuan CM. Regional heterogeneity in the distal motor axon: three zones with distinctive intrinsic components. J. Neurocytol. 1988 ; 17:649– 56 Yu J, Vodyanik MA, Smuga-Otto K, Antosiewicz-Bourget J, Frane JL, Tian S, Nie J, Jonsdottir GA, Ruotti V, Stewart R, Slukvin II, Thomson JA. Induced pluripotent stem cell lines derived from human somatic cells. Science. 2007 Dec 21; 318(5858):1917-20. Yu X, Duan KL, Shang CF, Yu HG, Zhou Z. Calcium influx through hyperpolarizationactivated cation channels (I(h) channels) contributes to activity-evoked neuronal secretion. Proc Natl Acad Sci U S A. 2004 Jan 27; 101(4):1051-6. Yutoku M, Grossberg AL, Pressman D. A cell surface antigenic determinant present on mouse plasmacytes and only about half of mouse thymocytes. J Immunol. 1974 May; 112(5):1774-81. Z Zammit P, Beauchamp J. The skeletal muscle satellite cell: Stem cell or son of stem cell. Differentiation 2001; 68:193–204. Zammit PS, Golding JP, Nagata Y, Hudon V, Partridge TA, Beauchamp JR. Muscle satellite cells adopt divergent fates: a mechanism for self-renewal? J. Cell Biol. 2004; 166:347–357 Zhang Y, Liu Y, Qu J, Hardy A, Zhang N, Diao J, Strijbos PJ, Tsushima R, Robinson RB, Gaisano HY, Wang Q, Wheeler MB. Functional characterization of hyperpolarization-activated cyclic nucleotide-gated channels in rat pancreatic beta cells. J Endocrinol. 2009 Oct; 203(1):45-53. Zhao H, Halicka HD, Li J, Biela E, Berniak K, Dobrucki J, Darzynkiewicz Z. DNA damage signaling, impairment of cell cycle progression, and apoptosis triggered by 5ethynyl-2'-deoxyuridine incorporated into DNA. Cytometry A. 2013 Nov; 83(11):97988. 164 Zhao T, Zhang ZN, Rong Z, Xu Y. Immunogenicity of induced pluripotent stem cells. Nature. 2011 May 13; 474 (7350):212-5. Zhong H, Jin Z, Chen Y, Zhang T, Bian W, Cui X, Jing N. First intron of nestin gene regulates its expression during C2C12 myoblast differentiation. Acta Biochim Biophys Sin (Shanghai) 2008; 40: 526–532. Zhou HM, Wang J, Rogers R, Conway SJ. Lineage-specific responses to reduced embryonic Pax3 expression levels. Dev Biol. 2008; 315: 369–82. Zhou S, Morris JJ, Barnes Y, Lan L, Schuetz JD, Sorrentino BP. Bcrp1 gene expression is required for normal numbers of side population stem cells in mice, and confers relative protection to mitoxantrone in hematopoietic cells in vivo. Proc Natl Acad Sci USA 2002; 99:12339-12344. Zhou S, Schuetz JD, Bunting KD, Colapietro AM, Sampath J, Morris JJ, Lagutina I, Grosveld GC, Osawa M, Nakauchi H, Sorrentino BP. The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype. Nat Med 2001; 7:1028-1034. Zhou T, Benda C, Duzinger S, Huang Y, Li X, Li Y, Guo X, Cao G, Chen S, Hao L, Chan YC, Ng KM, Ho JC, Wieser M, Wu J, Redl H, Tse HF, Grillari J, GrillariVoglauer R, Pei D, Esteban MA. Generation of induced pluripotent stem cells from urine. J Am Soc Nephrol. 2011 Jul; 22(7):1221-8.
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2.2.3 Résolution
La résolution de l'équation différentielle (2.29) dans le cas général (forces d'excitations arbitraires) n'étant pas pertinente, seuls les cas rencontrés en pratique sont étudiés. La résolution se limitera donc aux deux méthodes d'excitation suivantes : – cas d'une force ponctuelle en bout de poutre – cas d'une pression uniformément répartie sur la longueur de la poutre Ces deux méthodes correspondent aux deux méthodes classiquement utilisées au laboratoire IMS, à savoir un actionnement électromagnétique et un actionnement par céramique piézoélectrique.
2.2.3.1 Cas d'une force ponctuelle en bout de poutre
L'actionnement électromagnétique consiste à faire circuler un courant électrique dans une piste conductrice placée en bout de poutre. La présence d'un champ magnétique va ainsi entraîner l'apparition d'une force de Laplace localisée en bout de poutre (voir figure 2.2 page suivante). Cette force ponctuelle Fmag (ω) se traduit par une contrainte en cisaillement sur la face en bout de poutre. On a donc :
3 Fmag (
ω
)
∂
W
(
x, ω) = (2.33) 3 ∂x ÊI x=L1
L'équation à résoudre devient donc
:
ÊI ∂4 W (x, ω) + iωξ(ω)W (x, ω) − ω 2 μW (x, ω) = Fhydro (x, ω) ∂x4 (2.34) 48 z y Courant I x h1 L1 Champ B b
1 Fig. 2.2: Micropoutre actionnée par une force ponctuelle à son extrémité, exemple d'une force de Laplace où Fhydro (x, ω) est la force par unité de longueur exercée par le fluide le long de la poutre donnée par le modèle de Sader. Et les conditions aux limites sont l'équation (2.33) et : W (0, ω) = 0 ∂
(x, ω) =0 ∂x x=0 2 ∂ W (x, ω) =0 ∂x2 x=L1 Après résolution, on obtient l'expression analytique de la déformée en fonction de x et ω : W (x, ω) = Fmag (ω)L31 1 Φmag (x, ω) 2λ3 ÊI Rλ avec : Rλ = cos λ + 1 cosh λ (2.38) (2.39) 49 2.2. Résolution de l'équation différentielle du mouvement et λx λx sin
λ Φmag (x, ω) = cos − cosh + tanh λ L1 L1 cosh λ λx λx cos λ + sinh − sin +1 L1 L1 cosh λ où λ = L1 −iωξ(ω) + ω 2 μ + π4 ω 2 ρ0 b21 Γrect (ω) (2.40) 14 (2.41) ÊI
On peut remarquer que l'amplitude de vibration est, depuis la prise en compte des pertes, déterminée grâce aux conditions aux limites. En revanche, la fréquence de résonance n'est plus directement calculable. Ceci provient du fait que λ est complexe et n'annule donc plus l'expression (2.39). Cette particularité impose une étude spécifique pour la détermination de la fréquence de résonance et fait l'objet de la section 2.2.4.
2.2.3.2 Cas d'une pression uniforme répartie
L'actionnement par céramique piézoélectrique consiste à faire bouger le support de la poutre à la fréquence d'excitation voulue (voir figure 2.3 page suivante). Le mouvement ainsi généré interdit de considérer le référentiel associé au support comme Galiléen. L'étude du mouvement de la poutre dans le référentiel associé au support doit donc faire intervenir la force d'inertie due au mouvement du support. Si on considère un déplacement z(t) selon l'axe z, la force d'inertie dfi (t) qui s'applique à un élément de masse dm s'exprime par : dfi (t) = d2 z(t) dm dt2 Ce qui, autrement écrit, donne : 2 d z(t) dfi (t) = ρ1 h1 (b1 dL1 ) dt2 (2.42) (2.43) où dL1 est la longueur de l'élément de masse considéré. On que bien ainsi que le mouvement du support se traduit en une pression uniformément répartie sur toute la surface de la poutre (surface L1 b1 ). En appliquant la force d'inertie sur des volumes de longueur unitaire, l'équation à résoudre devient :
ÊI ∂4 W (x, ω) + iωξ(ω)W (x, ω) − ω 2 μW (x, ω) = Fhydro (x, ω) + Finertie ∂x4 (2.44) 50 z y x L1 h1 b1
Céramique piézoélectrique d(t) Fig. 2.3: Micropoutre actionnée par une céramique piézoélectrique 2.2. 2.2.4 Méthode d'extraction de la fréquence de résonance et du facteur de qualité
À partir des expressions (2.38) et (2.50) des déformées, une étude simple peut être menée pour approcher la fréquence de résonance. 52 En effet, quand λ est réel, le terme Rλ s'annule pour des valeurs particulières λn solutions de l'équation (1.12). Ceci correspond au cas où aucune perte n'est présente et traduit ainsi la divergence du fait de la résonance et de la présence d'une force d'excitation. En revanche, quand λ est complexe mais à partie imaginaire proche de 0 (donc en présence de faibles pertes), le terme Rλ ne s'annule plus mais est minimum pour certaines valeurs de λ. La résonance ne se traduit ainsi plus par une divergence et ne peut donc plus être déterminée aussi facilement. Toutefois, compte tenu de la forme des expressions (2.40) et (2.51) dictant l'allure de la déformée le long de la poutre, il est possible d'observer qu'aucune variation brutale ne se produit quand λ « tend 4 » vers λn. Ainsi, le comportement en fonction de λ des expressions des déformées est principalement dicté par le comportement du terme Rλ autour de λn. En effet, autour de λn, le terme | R1λ | possède un maximum quand |λ| tend vers λn avec λ à partie imaginaire proche de 0. Ceci entraîne qu'il est possible d'obtenir une bonne approximation de la fréquence de résonance en présence de faibles pertes en résolvant l'équation : |λ| = λn (2.52) En revanche, la détermination du facteur de qualité à un mode de résonance ne peut pas se faire en se limitant à l'étude de λ. Il est donc judicieux de déterminer le facteur de qualité en analysant la courbe de la réponse fréquentielle autour de la résonance. Le facteur de qualité est déduit grâce à l'expression suivante : Q= f0 ∆f (2.53) où f0 est la fréquence de résonance et ∆f la largeur de bande où l'amplitude en module vaut √12 fois l'amplitude en module à la résonance. Toutefois, cette méthode de détermination du facteur de qualité, bien que systématique, possède deux, légers, inconvénients. Premièrement, le facteur de qualité déduit est imprécis dans le cas des fortes pertes voire indéterminable lorsque le pic de résonance n'est pas assez prononcé. 2.3 Afin d'observer l'influence de l'excitation sur la réponse de la poutre, il est possible de tracer la réponse fréquentielle de la poutre pour les deux modes d'excitation proposés. Il est possible de montrer, par l'exemple, l'influence du mode d'excitation sur la réponse fréquentielle. Pour cela, on choisit une poutre fabriquée en silicium mono-cristallin orienté h100i, baignant dans l'air à température ambiante (T = 300K) et à pression atmosphérique P = 1bar, et de dimensions L1 = 100 μm, b1 = 30 μm et h1 = 5 μm. Le matériau considéré pour la poutre a pour caractéristiques principales : – Ê1 = 133,6 GPa – ν1 = 0,279 – ρ1 = 2 330 kg/m3 – α1 = 2,6 10−6 /K – Cp1 = 700 J/kg * K – κ1 = 150 W/m * K Le fluide considéré (l'air) a pour caractéristiques principales : – ρ0 = 1,179 kg/m3 – η0 = 18,57 10−6 Pa * s – c0 = 347,6 m/s
2.3.1 Déformée en bout de poutre et moment fléchissant à l'encastrement
Les deux modes d'excitation envisagés (électromagnétique et céramique piézoélectrique) sont comparés en terme d'influence sur la déformée en bout de poutre et sur le moment fléchissant à l'encastrement. Afin de faciliter la comparaison graphique des deux modes d'excitation, la déformée dans le cas de l'excitation par céramique piézoélectrique sera déterminée en considérant que la force d'inertie est indépendante de la fréquence. En effet, la résolution faite dans la section précédente fait apparaître un terme ω 2 lié au fait que l'excitation est une force d'inertie. On considérera 54 donc une d'excitation indépendante de la fréquence afin de faire disparaître l'influence du terme ω 2 afin de pouvoir comparer les comportements de façon graphique.
2 Flèche (normalisée) en bout de poutre 10 0 10 -2 10 -4 10 Actionnement magnétique Actionnement par céramique -6 10 5 10 6 10 Fréquence d'excitation (Hz) 7 10
Fig. 2.4: Flèche (en module) en bout de poutre normalisée à l'amplitude du premier mode de vibration, pour les deux modes d'actionnement
Les figures 2.4 et 2.5 illustrent le comportement fréquentiel d'une micropoutre selon le mode d'excitation. Il est ainsi possible de remarquer que le comportement général est le même quel que soit le mode d'excitation et plus particulièrement à la résonance. En dehors de la résonance, il est possible de remarquer l'existence d'antirésonances pour la flèche en bout de poutre entre les modes de vibration. De plus, ces anti-résonances ne se produisent pas à la même fréquence d'excitation selon le mode d'excitation, illustrant ainsi l'influence de l'excitation sur le comportement de la poutre en dehors de la résonance. En revanche, aucune anti-résonance n'est présente dans le cas de l'actionnement magnétique si on observe les contraintes normales à l'encastrement, alors que des anti-résonances existent lors d'une excitation par céramique piézoélectrique.
2.3. Réponse fréquentielle
0 Contraintes normales (normalisées) à l'encastrement 10 -2 10 -4 10 -6 10 Actionnement magnétique Actionnement par céramique -8 10 5 10 6 10 Fréquence d'excitation (Hz) 7 10
Fig. 2.5: Contraintes normales (en module) à l'encastrement normalisées à l'amplitude du premier mode de vibration, pour les deux modes d'actionnement
2.3.2 Mesure par piézorésistance
Afin d'obtenir un signal représentatif du mouvement de la poutre, la méthode de mesure choisie au laboratoire, dans un souci de miniaturisation et d'intégration, est une mesure piézorésistive. L'idée principale d'une telle mesure provient du fait que le silicium dopé est un matériau piézorésitif, c'est-à-dire un matériau dont la résistivité dépend des contraintes appliquées. Ainsi, en plaçant une piézorésistance à l'endroit où les contraintes normales (selon l'axe x) générées par le mouvement de la poutre sont maximales il est possible d'obtenir un signal électrique dont les variations sont liées au mouvement de la poutre. Cette piézorésistance sera donc réalisée lors du procédé de fabrication de la poutre dopage de bore dans le silicium. 2.3.2.1 Dans le cas des faibles déflexions, les contraintes normales selon l'axe x σxx sont liées au moment fléchissant Mx dans la section d'abscisse x par la 56 relation : Mx hp (2.54) I où hp est la distance à l'axe neutre à laquelle est placée la piézorésistance, et avec : ∂2w Mx = −ÊI 2 (2.55) ∂x On obtient donc : ∂2w σxx = −hp Ê 2 (2.56) ∂x Si on ne tient compte que des contraintes normales selon l'axe x, la dépendance aux contraintes normales d'un élément de résistance de longueur dl à l'abscisse l est donnée par : σxx = − dRpiezo = R (1 + πL σxx (l)) dl bpiezo (2.57) où R est la résistance par carré5, πL est le coefficient piézorésistif longitudinal et bpiezo la largeur de la résistance. 2.3.2.2 Influence de la longueur de la piézorésistance
Si on trace la courbe représentant le moment fléchissant le long de la poutre pour les premiers modes, on peut justifier le fait de placer la piézorésistance à l'encastrement de la poutre (voir figure 2.6). En effet, le moment fléchissant est maximal à l'encastrement et donc la variation de résistance sera maximale. De plus, il faut être prudent sur le choix de la taille de la piézorésistance. En effet, il est facile d'observer que si la résistance est très longue, la somme des variations de résistance le long de la poutre peut s'annuler pour certains modes de résonance. De façon grossière, une règle simple pourrait stipuler que pour observer le mode d'ordre n le rapport entre la longueur de la poutre et la longueur de la résistance ne doit pas être un multiple de n − 1. Par exemple, une résistance d'environ la moitié de la longueur de la poutre ne peut pas observer le mode 3. 5 donnée technologique dépendante du procédé de fabrication indiquant la valeur d'une résistance implantée dont la longueur est égale à sa largeur 57
2.4
.
Optimisation du facteur
de
qualité 0,15 Moment fléchissant (normé) 0,1 0,05 0 -0,05 -0,1 -0,15 -0,2 -0,25 0 Mode 1 Mode 2 Mode 3 0,1 0,2 0,3 0,4 0,5 0,6 0,7 0,8 0,9 1 Abscisse relative (x/L1)
Fig. 2.6: Moment fléchissant (normé) le long de la poutre pour les trois premiers modes de résonance en l'absence de pertes
2.4 Optimisation du facteur de qualité
Lorsqu'on exprime le facteur de qualité total en tenant compte de toutes les pertes, il est possible d'observer, à matériau et fluide donnés, qu'en fonction de la géométrie certaines pertes sont dominantes par rapport aux autres. Ainsi, il est possible de remarquer que : – les tes visqueuses sont dominantes pour des grandes valeurs de Lh11 ou pour des largeurs b1 très faibles – les pertes à l'encastrement sont dominantes pour des grandes valeurs de Lh11 – les pertes acoustiques sont dominantes pour des grandes largeurs ou pour des poutres proches de la coïncidence acoustique (nombre d'onde dans le milieu proche du nombre d'onde structurel) – les pertes internes sont généralement faibles mais maximales pour une √ valeur particulière de h1 ω. À partir de ces constatations, il est possible de voir qu'il n'y a, a priori, pas de géométrie idéale pour minimiser toutes les pertes en même temps. En revanche, à longueur donnée il existe un couple épaisseur - largeur qui maximise le facteur de qualité. En effet, il semble intuitif de constater qu'il peut exister un rapport Lh11 pour lequel les contributions des pertes visqueuses et des pertes à l'encas- 58 trement sont comparables, et qu'il peut exister une largeur pour laquelle les contributions des pertes visqueuses et des pertes acoustiques sont comparables. À matériau et à fluide donné, en déterminant le facteur de qualité pour une longueur de poutre donnée on peut observer qu'il existe bien une largeur et une épaisseur optimale en terme de facteur de qualité. La figure 2.7 représente le comportement du facteur de qualité en fonction de l'épaisseur et de la largeur pour une poutre de longueur L1 = 100μm. On peut ainsi constater les zones dans lesquelles certaines pertes dominent et remarquer ainsi que pour des rapports de forme classiques ( Lh11 > 100 et L1 > 10) la perte dominante est liée à la viscosité du fluide (l'air ici).
b1 -4 10 1200 Pe r te 1000 sa Pertes à l'encastrement co ti q ue s Épaisseur (m) us 800 -5 10 600 Pe rte -6 10 -6 10 400 sv isq u eus es -5 10 200 -4 10 Largeur (m) -3 10
Fig. 2.7: Facteur de qualité (iso-valeurs) à L1 = 100μm en fonction de la largeur et de l'épaisseur
Il est possible d'observer plus en détail l'influence de chaque perte autour de cet optimum et de constater que l'épaisseur optimale est principalement dictée par la combinaison des pertes visqueuses et des per à l'encastrement (voir figure 2.8) et que la largeur optimale est principalement dictée par les pertes visqueuses et dans une moindre mesure par les pertes par rayonnement acoustique (voir figure 2.9). De plus, on peut aussi remarquer que les pertes thermoélastiques n'ont qu'un effet modéré sur la localisation de l'optimum.
2.4. Optimisation du facteur de qualité
6 10 5 10 Total Acoustique Encastrement Thermoélastique Visqueux 4 10 3 10 2 10 -6 10 -5 10 Épaisseur de poutre (m) -4 10
Fig. 2.8: Dépendance à l'épaisseur d'une poutre de 100 μm de long à la largeur optimale (26,9 μm) 5
10 4 10 Total Acoustique Encastrement Thermoélastique Visqueux 3 10 2 10 -6 10 -5 -4 10 10 Largeur de poutre (m) -3 10
Fig. 2.9: Dépendance à la largeur d'une poutre de 100 μm de long à l'épaisseur optimale (7,46 μm)
60 Modélisation sans couche : influence des pertes
Ainsi, il est possible de classer les pertes par ordre d'importance : 1. pertes visqueuses 2. pertes à l'encastrement 3. pertes par rayonnement acoustique 4. pertes thermoélastiques Ces résultats, illustrés ici dans un cas précis, se confirment quelle que soit la longueur considérée. Il est ainsi possible de déterminer les épaisseurs et largeurs optimales pour différentes longueurs de poutre et de construire une courbe donnant les dimensions optimales à longueur de poutre donnée. Plus précisément, il est plus judicieux de construire une courbe donnant les rapports de forme Lb11 et Lh11 pour différentes longueurs (voir figures 2.10 et 2.11).
12 8 1 Rapport L /b 1 10 6 4 2 1 0 -7 10
Fig. 2.10: Rapport de poutre L1 b1 -6
10
-5 10 Longueur (m) -4 10 -3 10
à
fa
cteur de qualité optimal pour différentes longueurs
À partir de ces courbes, il est ainsi possible d'énoncer une règle simple pour déterminer de façon approchée la largeur et l'épaisseur d'une poutre de longueur donnée. En effet, on peut constater qu'utiliser les rapports de forme L1 = 3 et Lh11 = 10 permet de s'approcher simplement de l'optimum de facteur b1 de qualité. Ces rapports de forme particuliers ont l'avantage de proposer un facteur de qualité pour lequel la part des
2.4. Optimisation du facteur de qualité
16 12 1 Rapport L /h 1 14 10 8 6 4 -7 10
Fig. 2.11: Rapport de poutre L1 h1 -6
10
-5 10 Longueur (m) -4 10 -3 10
à facteur de qualité optimal pour différentes longueurs acoustique est moindre, rendant ainsi le résultat moins imprécis dans le cas d'erreurs dues au calcul des pertes acoustiques. De plus, cette règle simple est aussi intéressante du point de vue de la fréquence de résonance car la sensibilité augmente avec la fréquence de résonance. 2.5 Influence de la température et de la pression
À partir de l'équation (2.41) et de la méthode d'extraction de la fréquence de résonance proposée en section 2.2.4, il est possible d'avoir une bonne approximation du comportement de la micropoutre, et donc, plus particulièrement, de bien prédire le comportement vis-à-vis des variations de température et de pression.
2.5.1 Dépendances des divers paramètres vis-à-vis de la température et de la pression
Il faut, dans un premier temps, déterminer les effets de la température et de la pression sur les divers paramètres géométriques et physiques. Ces effets interviennent à deux niveaux, sur la poutre et sur son environnement (l'air). La température agit sur les dimensions et sur la masse volumique du silicium par l'intermédiaire de la dilatation thermique et permet ainsi d'en exprimer leur dépendance pour de faibles variations de température.
L1 = L10 (1 + α1 (T − T0 )) b1 = b10 (1 + α1 (T − T0 )) h1 = h10 (1 + α1 (T − T0 )) ρ1 = ρ10 (1 − 3α1 (T − T0 ))
où α1 le coefficient de dilatation thermique et vaut 2,6 10−6 /K, T est la température, T0 la température de définition des paramètres, ρ10 la masse volumique du silicium à T = T0 et L10, h10 et b10 sont les dimensions à T = T0. La température agit aussi sur la rigidité du silicium et peut s'exprimer, pour de faibles variations de température, grâce à l'expression : E1 = E10 (1 + αE (T − T0 )) avec αE = −92 10−6 /K. 2.5. Influence de la température et de la pression
Si on assimile l'air à un gaz parfait, ses propriétés sont affectées par les variations de température et de pression. La viscosité absolue de l'air dépend ainsi de la température et sa masse volumique dépend de la température et de la pression. Dans ces conditions, on a : P T η 0 = ηA T + ηB ρ0 = Λρ (2.63) (2.64) où P est la pression du gaz et avec ηA = 4,9 nPa * s/K, ηB = 17,1 μPa * s et Λρ = 3,536 10−3 s2 * K/m2. 2.5.2
Étant donnée la forme de l'expression modélisant les pertes dues à la viscosité du fluide, il est difficile de prévoir analytiquement la dépendance de la fréquence de résonance vis-à-vis de la température et de la pression. En revanche, il est possible de déterminer quels paramètres influent majoritairement. En reformulant l'équation (2.41) il est possible d'obtenir : !2 − 14 2 s μ π 2
−
Q + 4
ρ
0
b1
= (Γ(ω
))
ÊI λn
1
+
ω
=
π 2 L1 μ + 4 ρ0 b1
<
(Γ(ω)) μ + π4 ρ0 b21 < (Γ(ω)) (2.65) où < et = représentent, respectivement, les fonctions partie réelle et partie imaginaire. Si, de plus, on considère que l'influence des pertes est négligeable, l'expression devient : 2 s λn ÊI ω= (2.66) π L1 μ + 4 ρ0 b21 < (Γ(ω)) qui, si on remplace I et μ par leurs expressions et en regroupant certains termes, devient : "s s # 2 Ê λ 1 n * ω= h1 (2.67) ρ0 b1
L
1 12
ρ1
1 +
π
4
ρ1h <
(
Γ(ω
)) 1 À partir de cette équation, il est possible d'étudier séparément les deux termes entre crochets. Le premier terme représente la fréquence de résonance 64 sans pertes et dans le vide, alors que le deuxième terme représente l'influence des effets inertiels dûs au fluide. En connaissant la dépendance des caractéristiques du matériau de la poutre vis-à-vis de la température, il est possible de remarquer que l'effet dominant (à plus de 97%) dans la dépendance à la température de la fré quence de résonance dans le vide est causé par les variations du module de Young, l'influence de la dilatation étant négligeable. La contribution des effets inertiels est, en revanche, plus délicate à déterminer. Néanmoins, en étudiant les caractéristiques de l'air, il est possible de constater que la variation des propriétés de l'air (viscosité absolue et masse volumique) est la principale cause de la modification des effets inertiels devant la variation des propriétés physiques de la poutre. La dépendance à la pression est, quant à elle, liée à la modification des propriétés de l'air et est donc principalement liée à la variation de la masse volumique de l'air, la viscosité absolue de l'air ne variant que très peu en fonction de la pression. 2.5.3 Confirmation par l'exemple
Pour illustrer plus précisément l'influence de la température et de la pression, une poutre est choisie arbitrairement. Si on choisit une poutre en silicium de longueur L1 = 100 μm de largeur b1 = 30 μm et d'épaisseur h1 = 5 μm résonant dans l'air à T = 300 K et P = 1 bar, sa fréquence de résonance est : f0 ≈ 610 588 Hz (2.68) Si on calcule la variation de fréquence associée à une variation de température en considérant la dépendance en température de tous les paramètres (module de Young, coefficient de Poisson, masse volumique, longueur, largeur, épaisseur, viscosité et masse volumique de l'air), on obtient une dépendance à la température de : f0 ∆f ≈ −12,1 * 10−3 * ∆T T (2.69) En revanche, si on ne considère que la dépendance du module de Young, du coefficient de Poisson et de la viscosité et masse volumique de l'air, on obtient une dépendance à la température de : ∆f f0 ≈ −12,4 * 10−3 * ∆T T (2.70) 65 Les deux valeurs obtenues confirment donc bien le fait qu'il est possible de négliger la dilatation du matériau de la poutre et de ne considérer que les variations de propriétés mécanique de la poutre et les variations des propriétés physiques de l'air. En étudiant plus précisément la dépendance à la température, il est possible de remarquer que la contribution des effets inertiels a une influence sur la variation de fréquence opposée à celle produite par la variation de module de Young. Ainsi, selon la géométrie de la poutre, il est possible d'avoir une dépendance de signe opposé à celle présentée par l'expression (2.69). Il est donc envisageable de voir une poutre dont la fréquence de résonance soit, au premier ordre, indépendante de la température. 2.6 Conclusion
Après une présentation des modèles existants pour décrire l'essentiel des pertes liées aux micropoutres résonant dans l'air, ce chapitre a présenté une méthode de résolution de l'équation différentielle du mouvement en présence de pertes et soumis à une excitation arbitraire. La méthode a ensuite été appliquée aux cas d'excitation rencontrés au laboratoire afin d'illustrer l'influence du mode d'excitation sur la réponse fréquentielle d'une micropoutre. L'étude des pertes en fonction de la géométrie d'une poutre a permis de mettre en évidence la possible optimisation des dimensions à longueur de poutre donnée de façon à maximiser le facteur de qualité. Une telle optimisation permet ainsi, à partir d'une longueur de poutre donnée, de choisir facilement l'épaisseur et la largeur de la poutre garantissant la meilleure stabilité de la mesure de la fréquence de résonance en fonctionnement oscillateur.
Chapitre 3 Modélisation : influence de la couche
Pour convertir les micropoutres en capteur chimique, il faut déposer à la surface de celles-ci une couche sensible capable de détecter les molécules concernées. La présence d'une telle couche vient modifier le comportement mécanique de la structure notamment du fait de sa masse et de sa rigidité. De plus, les couches utilisées étant majoritairement des polymères, le solide ne peut, en général, plus être modélisé par un simple solide élastique. En effet, ces matériaux sont très souvent des matériaux dans lesquels des pertes apparaissent lorsqu'ils sont sollicités. Ainsi, lorsqu'ils sont utilisés pour réaliser des capteurs chimiques à base de micropoutres, leurs propriétés viscoélastiques ont un impact important sur les caractéristiques du capteur. C'est pourquoi, dans ce chapitre, un modèle intégrant les aspects viscoélastiques couches sensibles est présenté en vu d'optimiser le capteur vis-à-vis de sa limite de détection. La sensibilité et le facteur de qualité des poutres sont étudiés et plusieurs approches d'optimisation sont détaillées. 3.1 Optimisation de la sensibilité
Afin d'augmenter la sensibilité des capteurs, il existe deux principales approches comme vu dans la section 1.3.6.2. La première consiste à augmenter la fréquence de résonance et la deuxième consiste à augmenter l'épaisseur de couche. Ces deux approches sont étudiées dans les sections suivantes en gardant à l'esprit l'influence probable de la couche sensible.
3.1.1 Réduction de la longueur ou utilisation d'un mode d'ordre supérieur 3.1.1.1 Problématique
La réduction de la longueur ou l'augmentation de l'ordre du mode permettent dans les deux cas d'augmenter la fréquence de résonance. Ces deux méthodes peuvent être comparées car, comme le fait apparaître l'équation (1.21), rappelée ci-après, utiliser un mode de résonance plus élevé revient à réduire la longueur de la poutre.
v u h Ê 1u ωn = λ2n 2 t (1.21) L1 12 ρ + ρ h2 1 2 h1
L'étude proposée ici consiste à comparer les deux méthodes, équivalentes en terme de sensibilité, afin de déterminer s'il y en a une meilleure en terme de bruit de mesure et donc en terme de facteur de qualité. En effet, toutes les pertes considérées n'ont pas les même dépendances vis-à-vis de l'ordre du mode de résonance, de la géométrie ou de la fréquence. Il faut donc étudier plus précisément les impacts respectifs des deux méthodes. En observant les expressions permettant de calculer les différentes pertes, il est possible de comparer les deux méthodes lorsqu'elles entraînent la même augmentation de fréquence.
3.1.1.2 Pertes visqueuses
En observant l'expression (2.9), rappelée ci-après, il est possible de remarquer que ni la longueur ni l'ordre du mode n'interviennent et que le la fréquence de résonance intervient. Il n'y aura donc pas de différence en terme de pertes visqueuses entre une augmentation du mode et une réduction de la longueur du moment que la fréquence de résonance est identique.
Fhydro (x, ω) = 3.1.1.3 π ρ0 ω 2 b21 Γ(ω)W (x, ω) 4 (2.9)
Pertes internes
À partir de l'expression (2.19), rappelée ci-après, il est possible de remarquer que, comme pour les pertes visqueuses, seule la fréquence intervient. Il n'y a donc pas de différence en terme de pertes internes entre les deux méthodes. 3.1.1.5 R L1 1024πμω x=0 W 2 (x, ω)dx = R R π L1 ρ0 c0 (k0 b1 )4 θ=0 sin3 θ| x=0 W (x, ω) exp−ixk0 cos θ dx|2 dθ (2.28)
Pertes à l'encastrement
En ce qui concerne les pertes à l'encastrement, le raisonnement est moins immédiat car il dépend des modes considérés. À partir de l'équation de la fréquence de résonance, il est facile de montrer qu'utiliser le mode 2 au lieu du mode 1 est équivalent à réduire la longueur par un facteur d'environ 2,5. Ainsi, un rapide calcul de l'expression (2.18), rappelée ci-après, montre que réduire la longueur fait beaucoup plus chuter le facteur de qualité qu'utiliser un mode d'ordre supérieur.
2 3 L1 1 cos λn + cosh λn (2.18) Qenc = 3,966 λn sin λn − sinh λn h1
De façon plus générale, il est possible de remarquer qu'il vaut mieux, pour les pertes à l'encastrement, utiliser les modes d'ordre supérieur.
3.1.1.6 Effet global
Globalement, au vu des dépendances de chaque perte par rapport aux deux méthodes étudiées, l'utilisation de modes d'ordres supérieur est à privilégier par rapport à la réduction de la longueur. Ceci est d'autant plus intéressant que, la tendance actuelle étant de réduire les dimensions, il peut s'avérer plus délicat de réaliser les dépôts de couche sensible de façon fiable lorsque les dimensions se réduisent. Toutefois, cette étude ne permet pas de savoir si augmenter la fréquence par l'une ou l'autre de ces deux méthodes est préférable en terme de pertes. 70 En effet, bien que la fréquence de résonance soit plus élevée et que la sensibilité soit augmentée, il est possible que les pertes soient plus importantes et que, globalement, la limite de détection soit moins bonne. 3.1.2 Augmentation de l'épaisseur de couche sensible
À partir de l'expression (1.26), il est possible d'étudier l'influence de l'épaisseur de couche sensible sur la sensibilité. L'expression de la sensibilité peut s'écrire sous la forme : S Cg λ2 1 = n2 √ L1 8π 3 3 q Ê1 K h12 h2 3 (3.1) (ρ1 h1 + ρ2 h2 ) 2 Cette expression fait apparaître un terme noté γ dépendant des épaisseurs et des masses volumiques des matériaux : 3 γ= h12 h2 3 (3.2) (ρ1 h1 + ρ2 h2 ) 2 L'étude du terme γ permet de mettre en évidence l'existence d'un optimum théorique d'épaisseur de couche sensible (voir figure 3.1). Cet optimum h2opt est donné par l'expression : h2opt = 2 ρ1 h1 ρ2 (3.3) À cet optimum le terme γ vaut γopt qui s'exprime par : 2 h1 γopt = √ √ 3 3 ρ2 ρ1 (3.4) Ainsi il est possible de constater que, tant qu'une épaisseur de couche sensible optimale est choisie, la sensibilité augmente avec l'épaisseur de la poutre. Toutefois, l'épaisseur de poutre ne peut pas indéfiniment être augmentée, notamment pour les raisons de validité du modèle qui imposent une épaisseur plus petite que la longueur. On ne peut donc que constater que, pour optimiser les épaisseurs, il faut choisir une poutre d'épaisseur relativement importante, c'est-à-dire un rapport Lh11 le plus petit possible et ajuster l'épaisseur de couche sensible en conséquence en utilisant l'expression (3.3). Cette optimisation n'est toutefois valable que pour des couches sensibles dont le module de Young est très faible par rapport au module de Young du matériau utilisé pour la tre, c'est-à-dire lorsque la présence de la couche sensible ne vient pas modifier la raideur de la structure.
de la sensibilité
Sensibilité normalisée à 1 à l'optimum 1 0,8 0,6 0,4 0,2 0 -2 -1 0 1 2 10 10 10 10 10
Épaisseur de couche sensible normalisée à 1 à l'optimum Fig. 3.1: Sensibilité normalisée à 1 à l'optimum en fonction de l'épaisseur de couche sensible normalisée à 1 à l'optimum
De plus, cet optimum se traduit, vu les matériaux utilisés classiquement (silicium pour la poutre, polymères pour la couche sensible), par une expression approchée : h2opt ≈ 4h1 (3.5) Un tel optimum suppose donc une épaisseur de couche très importante par rapport à l'épaisseur de la poutre et peut avoir deux effets. 3.1.3 Les deux sections précédentes ont montré l'influence de la longueur et de l'épaisseur de façon indépendante et il a été montré qu'il fallait privilégier l'utilisation d'ordres supérieurs, les faibles longueurs et les fortes épaisseurs. 72 Ces deux conclusions ont pour effet de privilégier l'utilisation de poutres dont le rapport Lh11 est relativement faible, par exemple proche de 10. Toutefois, aucune indication quant aux ordres de grandeurs n'est donnée et c'est pourquoi l'étude de l'influence du facteur d'échelle est intéressante. Cette étude permet, à rapports de forme constants ( Lh11, Lb11 et hh21 ), de voir l'influence d'une réduction ou d'une augmentation des dimensions. À partir de l'expression (3.1), et en posant Lh11 = r et h2 = 2 ρρ21 h1, on obtient l'expression : s S Cg = λ2n 36π Ê1 1 1 K r ρ1 ρ2 L 1 (3.6) On constate ainsi que la réduction des dimensions, à rapports de forme constants et à épaisseur optimale de couche sensible, permet d'augmenter la sensibilité et la fréquence de résonance. 3.2 3.2.1 Modélisation avec viscoélasticité Les polymères et la viscoélasticité
La modélisation utilisée jusqu'à présent repose principalement sur la loi de Hooke où les contraintes sont toujours directement proportionnelles à l'élongation relative. Or, certains solides ne répondent pas parfaitement à cette loi. En particulier certains matériaux soumis à une élongation constante ne maintiennent pas une contrainte constante dans le temps. Ainsi, de tels matériaux soumis à des élongations sinusoïdales, ne génèrent pas de contraintes parfaitement en phase ni parfaitement en quadrature. Par conséquent une partie de l'énergie fournie au matériau est restituée et une partie est dissipée. Malgré tout, ces matériaux, classés comme viscoélastiques, peuvent être étudiés en utilisant une modélisation très similaire à celle utilisée jusqu'à présent, en leur associant un module de Young complexe dont la partie imaginaire reflète l'énergie dissipée. Les polymères, de par leur structure, sont de bons exemples de matériaux viscoélastiques. En effet, les chaînes de monomères peuvent se déformer de deux façons. À échelle microscopique, les chaînes sont élastiques, mais à plus grande échelle les chaînes peuvent se réorganiser en se déroulant, par exemple. L'objet de cette section consiste donc à proposer un modèle permettant de modéliser le comportement d'une poutre élastique recouverte par un polymère viscoélastique. Grâce à un tel modèle il devient possible de mieux prédire l'impact des pertes et donc du bruit sur la limite de détection.
3.2. Modélisation avec viscoélasticité 3.2.2 Modèle de base
Le modèle de base, qui est décrit dans cette section, a été développé en collaboration avec Marquette University [Sam06] [Duf07] et repose sur la détermination de rigidité d'une structure composée d'une poutre en matériau élastique (le silicium par exemple) recouverte par une couche viscoélastique (voir figure 3.2). E'2+iE''2 h2 hn h1 E1 L1
Fig. 3.2: Vue de côté d'une poutre recouverte de sa couche sensible
Ce modèle repose sur la description des matériaux par leur module de Young. Dans le cas de la couche viscoélastique, le module de Young considéré comporte une partie imaginaire reflétant les pertes.
3.2.2.1 Hypothèses, approximations
Les hypothèses considérées sont celles présentées dans la section 1.3.1 dans le but d'utiliser un modèle en 1-dimension. À partir de ces hypothèses et en considérant un mouvement sinusoïdal, il est possible d'établir les relations temporelles entre contraintes normales et allongement relatif pour la poutre et la couche viscoélastique. Ainsi, en supposant que l'ensemble des contraintes normales à une abscisse x donnée ne génère pas de force axiale (orientée selon la longueur de la poutre), il est possible d'établir la position de l'axe neutre. L'expression ainsi obtenue fait apparaître, dans le cas général, une position de l'axe neutre variant au cours d'une période d'oscillation. Toutefois, cette dépendance avec le temps étant relativement peu importante, il est possible d'approximer la position de l'axe neutre par sa valeur moyenne au cours du temps. 74 On obtient donc l'expression de la position moyenne de l'axe neutre par rapport à la surface libre de la couche sensible : h2 h1 E1 (h1 + h2 ) (h1 E1 + h2 E20 ) hn = + 2 2 (h1 E1 + h2 E20 )2 + h21 E2002 (3.7) où h1 est l'épaisseur de la poutre, h2 est l'épaisseur de la couche, E1 est le module de Young du matériau de la poutre, et E20, E200 sont, respectivement, la partie réelle et la partie imaginaire du module de Young complexe du matériau de la couche sensible à la fréquence considérée.
3.2.2.2 Rigidité équivalente de la poutre
À partir de la connaissance de la position de l'axe neutre, il devient possible d'établir la rigidité équivalente de la structure à partir du calcul du moment fléchissant. On obtient ainsi l'expression de la rigidité équivalente EI ∗ :
EI ∗ = E1 I1 + E20 I2 + iE200 I2 avec : 2 b1 h31 h1 I1 = + b1 h1 hn − h2 − 12 2 2 h2 b1 h32 + b1 h2 hn − I2 = 12 2
(
3.8)
(
3.9)
(3.10) Cette rigidité équivalente permet ainsi de résoudre l'équation différentielle du mouvement d'une poutre homogène, par l'intermédiaire de l'équivalence entre une poutre élastique recouverte d'un matériau viscoélastique et une poutre viscoélastique homogène, en considérant la rigidité équivalente EI ∗ et en tenant compte de la masse de la couche. Pour cela, on utilise les expressions suivantes : EI = EI ∗ (3.11) et μ = μ12 = ρ1 b1 h1 + ρ2 b1 h2 (3.12)
75
3.2. Modélisation avec viscoélasticité 3.2.3 Poutre totalement recouverte 3.2.3.1 Fréquence de résonance et facteur de qualité
À partir de l'équivalence exposée dans la section précédente, il est possible de résoudre l'équation différentielle du mouvement d'une poutre recouverte d'une couche sensible viscoélastique en l'absence d'autres pertes. La résolution permet d'extraire la fréquence de résonance de la structure ainsi que le facteur de qualité. On obtient ainsi [Sam05] les expressions de la fréquence de résonance fcouche : 2 s 1 λn E1 I1 + E20 I2 (3.13) fcouche = 2π L1 μ12 et du facteur de qualité associé aux pertes viscoélastiques Qcouche : Qcouche = 2 1− 3.2.3.2 1 q E200 I2 E1 I1 +E20 I2 (3.14) Pour illustrer la dépendance du facteur de qualité à l'épaisseur de couche sensible on considère une poutre fabriquée en silicium mono-cristallin orienté h100i, baignant dans l'air à température ambiante (T = 300K) et à pression atmosphérique P = 1bar, et de dimensions L1 = 100 μm, b1 = 30 μm et h1 = 5 μm. Le matériau considéré pour la poutre a pour caractéristiques principales : – Ê1 = 133,6 GPa – ν1 = 0,279 – ρ1 = 2 330 kg/m3 – α1 = 2,6 10−6 /K – Cp1 = 700 J/kg * K – κ1 = 150 W/m * K Le fluide considéré (l'air) a pour caractéristiques principales : – ρ0 = 1,179 kg/m3 – η0 = 18,57 10−6 Pa * s – c0 = 347,6 m/s 76 La poutre est totalement recouverte par une couche sensible qui est un polymère, le PIB (polyisobutylène), et dont les propriétés viscoélastiques (E20 et E200 ) dépendent de la fréquence. Les données considérées sont données par Ferry [Fer04] et sont tracées en figure 3.3. Les autres caractéristiques utiles sont : – sa masse volumique, ρ2 = 917 kg/m3 – son coefficient de Poisson, ν2 = 0,49.
10 E'2 E''2 9 10 2 2 Modules de Young, E' et E'' (Pa) 10 8 10 7 10 6 10 5 10 4 10 -3 10 0 10 3 10 6 10 9 10
Pulsation, ω (rad.s-1) Fig. 3.3: Parties réelles et imaginaire du module de Young complexe du PIB en fonction de la pulsation de sollicitation
En utilisant l'expression (3.14), on obtient la figure 3.4. On peut remarquer que le facteur de qualité chute rapidement pour atteindre moins de 100 à partir de 3 μm d'épaisseur de couche sensible. La rapidité de cette chute s'explique principalement par la fréquence de résonance. En effet, à 611 kHz, la sollicitation du polymère génère beaucoup de pertes du fait d'un E200 élevé (E200 = 306 MPa). Pour des poutres avec d'autres géométries, résonant à plus basse fréquence, il serait possible d'observer deux phase de variation pour le facteur de qualité (voir figure 3.5). Durant une première phase, mettant en oeuvre des épaisseurs faibles, le facteur de qualité peut augmenter avec l'épaisseur de couche du fait d'un apport de masse plus rapide que l'apparition de pertes viscoélastiques. Ensuite, à épaisseur plus importante, les pertes viscoélastiques interviendraient de façon plus prononcée faisant ainsi chuter le facteur de qualité.
1000 900 800 700 600 500 400 300 200 100 0 -8 10 -7 -6 10 10 Épaisseur de couche sensible (m) -5 10
Fig. 3.2.4 Poutre partiellement recouverte
Étant donné que les pertes viscoélastiques sont dues à la sollicitation de la couche sensible, il semble intuitif que placer la couche seulement aux endroits de faible sollicitation devrait permettre de réduire les pertes viscoélastiques. Ce gain potentiel implique de placer la couche sensible seulement du côté de l'extrémité libre (voir figure 3.6). Or, un calcul simple permet de trouver que l'influence d'une masse additionnelle est d'autant plus importante qu'elle est proche de l'extrémité libre. Ces deux effets semblent donc bénéfiques et il semble judicieux de disposer d'un modèle décrivant l'influence de la couche en fonction de la couverture de la poutre. 3.2.4.1 Afin d'estimer le gain en terme de facteur de qualité, il est possible d'utiliser une approche simplifiée qui modélise une poutre partiellement recouverte par deux morceaux de poutres, le premier, proche de l'encastrement, peut être modélisé par le modèle présenté dans le chapitre précédent, tandis que le deuxième morceau, proche de l'extrémité libre, peut être modélisé par le modèle présenté dans la section précédente.
3.2.4.2 Résolution
La résolution est donc basée sur la résolution de deux équations différentielles modélisant le comportement des deux morceaux (avec et sans couche sensible). Les conditions initiales sont donc associées à trois abscisses le long de la poutre : à l'encastrement (x = 0), à l'extrémité libre (x = L1 ) et au niveau de la transition sans/avec couche (x = (1 − δ) L1 ). Plus précisément, ces conditions initiales sont : 79
3.2. Modélisation avec viscoélasticité z y x h2 h1 δ.L1 L1 b1 Micropoutre Fig. 3.6: Géométrie d'une micropoutre partiellement
couverte de couche sensible – à l'encastrement et à l'extrémité libre, celles utilisées jusqu'à présent (pas de mouvement à l'encastrement, pas de contraintes à l'extrémité libre). – au niveau de la transition, on considère qu'il y a continuité du mouvement et continuité des contraintes. La résolution analytique de l'équation différentielle, bien que sans difficultés particulières, permet d'obtenir une expression analytique de la déformée de la poutre en fonction de l'abscisse et de la fréquence. Toutefois, l'expression obtenue est d'une taille (en nombre de termes) qui rend difficile sa manipulation autrement que par l'utilisation d'un logiciel de calcul numérique. L'expression permet néanmoins de déterminer, numériquement, la fréquence de résonance et le facteur de qualité. 3.3 Optimisation en terme de limite de détection
La détermination des performances en terme de limite de détection d'une poutre faisant intervenir à la fois la sensibilité et le bruit de mesure, il est important, et judicieux, de prendre en considération la dépendance de la sensibilité et du facteur de qualité vis-à-vis de l'épaisseur de couche sensible. En effet, comme l'ont montré certaines études [Fad04] [Lan02], la sensibilité et le bruit augmentant avec l'épaisseur de couche sensible, il faut étudier l'effet global afin de déterminer l'impact sur la limite de détection.
3.3. Optimisation en terme de limite de détection
À partir de deux exemples, deux études sont proposées pour illustrer comment optimiser la limite de détection d'une poutre donnée en choisissant soit l'épaisseur de couche sensible à déposer sur toute la poutre, soit le taux de couverture à épaisseur de couche donnée. La poutre utilisée comme modèle est celle décrite en section 3.2.3.2, appliquée à la détection de toluène avec du PIB comme couche sensible dont le coefficient de partage vaut K = 583 [Jon05].
3.3.1 Épaisseur de couche optimale à poutre totalement recouverte
Cette première étude (basée sur les expressions présentées dans la section 3.2.3) illustre, par exemple, le cas d'une optimisation lorsqu'il n'est pas possible d'ajuster le taux de couverture de la poutre alors qu'il est possible d'ajuster l'épaisseur de couche sensible déposée. L'augmentation de l'épaisseur de couche sensible entraînant, comme vu précédemment, la diminution du facteur de qualité (voir figure 3.4) mais entraînant aussi l'augmentation de la sensibilité (voir figure 3.8), il faut étudier la limite de détection, combinaison du bruit et de la sensibilité, définie comme la plus petite concentration détectable si suppose être capable de discerner un décalage de fréquence égal à « trois fois le bruit ». Décalage en fréquence (Hz) 0,07 0,06 0,05 0,04 0,03 0,02 0,01 0 -7 10 -6 10 Épaisseur de couche sensible (m) -5 10
Fig. 3.8: Décalage de fréquence, dû à la détection de 1 mg/m3 de toluène avec du PIB, en fonction de l'épaisseur de polymère pour une poutre de 100 μm de long, 30 μm de large et 5 μm d'épais 82
Le « bruit » considéré est le décalage de fréquence lié à une fluctuation de phase ψ(t) dans la boucle de l'oscillateur, donné par l'expression (1.32). ∆fbruit (t) = f0 ψ(t) 2Q (1.32) On peut ainsi écrire l'expression de la limite de détection, notée LOD : LOD = 3∆fbruit (t) S Cg (3.15) À partir de cette expression, on peut représenter la limite de détection en fonction de l'épaisseur de couche sensible, en la normalisant en un point afin de s'affranchir des termes inconnus (la fluctuation de phase de l'oscillateur, par exemple). Cette courbe (voir figure 3.9) fait apparaître une épaisseur optimale aux environs de 1,3 μm pour une poutre de 100 μm de long, 30 μm de large et 5 μm d'épais. À cet optimum le facteur de qualité vaut environ 280 alors que le facteur de qualité sans couche sensible vaut environ 976. Cette épaisseur optimale (h2 = 1,3 μm), est bien en deçà de l'épaisseur optimale en terme de sensibilité (h2 ≈ 25 μm) pour une couche qui serait très peu rigide. Un tel écart illustre bien la nécessité de prendre en compte les aspects concernant le bruit de mesure en plus de la sensibilité. Limite de détection normalisée 6 5 4 3 2 1 -7 10 -6 10 Épaisseur de couche sensible (m) -5 10
Fig. 3.9: Limite de détection, normalisée à 1 pour l'épaisseur optimale (1,3 μm), en fonction de l'épaisseur de polymère pour une poutre de 100 μm de long, 30 μm de large et 5 μm d'épais 83
3.3. Optimisation en terme de limite de détection
L'intérêt principal de cette étude est de montrer qu'il est possible de prédire théoriquement l'épaisseur de couche sensible à condition de connaître les propriétés viscoélastiques de la couche sensible dans la plage de fréquence d'utilisation. En l'absence de données sur la couche sensible, il faudrait se contenter d'une approche expérimentale qui consisterait à déposer différentes épaisseurs de couche sensible et à faire des mesures de bruit afin d'estimer au cas par cas la limite de détection.
3.3.2 Taux de couverture optimal à épaisseur de couche donnée
Cette étude (basée sur le calcul des expressions présentées dans la section 3.2.4) illustre le cas où il n'est pas possible d'ajuster l'épaisseur de couche (du fait de contraintes liées au procédé de dépôt, par exemple) mais qu'il est possible d'ajuster le taux de couverture (classiquement en cas de procédé par photo-lithographie). Comme précédemment, à partir du calcul du facteur de qualité (voir figure 3.7) et du décalage de fréquence (voir figure 3.10) en fonction du taux de couverture (la couche étant présente seulement à l'extrémité libre de la poutre), il est possible de déterminer le taux optimal de couverture afin de minimiser la limite de détection. Décalage en fréquence (Hz) 0,05 0,04 0,03 0,02 0,01 0 0 0,2 0,4 0,6 Taux de couverture 0,8 1
Fig. 3.10: Décalage de fréquence dû à la sorption de 1 mg/m3 de toluène en fonction du taux de couverture pour une poutre de 100 μm de long, 30 μm de large, 5 μm d'épais et couverte de 5 μm de PIB 84
On obtient ainsi la figure 3.11 sur laquelle il est possible d'observer que la couverture optimale est d'environ 0,32 soit 32% pour une poutre de 100 μm de long, 30 μm de large, 5 μm d'épais et couverte de 5 μm de PIB. À ce taux de couverture, le facteur de qualité vaut environ 666 alors que sans couche sensible il est d'environ 976 et qu'avec une couverture totale il ne dépasse pas 40. Limite de détection normalisée 20 15 10 5 0 0 0,2 0,4 0,6 Taux de couverture 0,8 1
Fig. 3.11: Limite de détection, normalisée à 1 pour le taux de couverture optimal, en fonction du taux de couverture pour une poutre de 100 μm de long, 30 μm de large, 5 μm d'épais et couverte de 5 μm de PIB
On peut comparer le facteur de qualité ainsi obtenu (666) avec le facteur de qualité obtenu dans la section précédente (280) où une couche plus fine était déposée sur la totalité de la poutre. De plus, le décalage de fréquence en sorption précédemment obtenu (environ 16,7 mHz) est nettement inférieur à celui obtenu avec une épaisseur de couche plus importante et un taux de couverture de 32% (environ 40,5 mHz). On constate que ne déposer la couche sensible que sur une partie de la poutre permet un gain significatif en terme de sensibilité, de facteur de qualité et donc de limite de détection.
85 3.3. Optimisation en terme de limite de détection 3.3.3 Vers la recherche d'un couple optimal taux de couverture - épaisseur de couche sensible
À partir de la poutre précédemment étudiée et en prenant en compte à la fois l'épaisseur de couche sensible et le taux de couverture, il est possible de montrer qu'il est intéressant, en terme de limite de détection, de localiser au maximum un volume important de couche sensible en bout de poutre (voir figure 3.12).
-5 Épaisseur de couche sensible (m) 10 1,1 1,4 2 -6 10 3 6 10 14 20 30 50 -7 10 100 -8 10 0,05 400 0,2 200 0,4 0,6 Taux de couverture 0,8 1
Fig. 3.12: Isovaleurs de la limite de détection, normalisée à 1 au minimum, en fonction du taux de couverture et de l'épaisseur de couche sensible, et lieu des points (ligne blanche) indiquant le taux de couverture optimal à épaisseur donnée. Les
leurs indiquées correspondent à la limite de détection normalisée On peut ainsi remarquer, qu'avec une épaisseur de couche sensible de 10 μm avec un taux de couverture de 0,27 le gain en terme de limite de détection est d'environ 3 par rapport à une couche sensible de 1 μm d'épaisseur avec un taux de couverture de 0,47. 3.4 Influence de la sorption sur les propriétés viscoélastiques
La sorption d'un gaz dans la couche sensible provoque principalement la modification de sa masse volumique, comme considéré jusqu'à présent, mais peut aussi provoquer une modification des propriétés viscoélastiques [Li05]. Deux effets ont lieu et proviennent du fait que les molécules de gaz interagissent avec les molécules de la couche sensible modifiant ainsi les interactions entre les molécules de la couche sensible. Il en résulte une modification des composantes E20 et E200 qui se traduisent généralement par une baisse de ridigité (E20 ) et une augmentation des pertes (E200 ). À partir des expression (3.7) à (3.10) il est possible de séparer les effets des variations de E20 et E200 sur les expressions (3.13) et (3.14) en trois phénomènes. Le premier concerne la modification directe de EI ∗ via la variation de E20, le second concerne la modification directe de EI ∗ via la variation de E200 et le troisième concerne la modification indirecte de EI ∗ causée par la modification de la position de l'axe neutre (et donc de I1 et I2 ) via les variations de E20 et E200. On peut ainsi écrire :
μ12 I2 ∆E20 E20 ∆I1 + E20 ∆I2 ∆fcouche + =− + fcouche μ12 2 (E1 I1 + E20 I2 ) 2 (E1 I1 + E20 I2 ) (3.16
) À partir des développements limités à l'ordre 1 des expressions (3.9) et (3.10), puis en supposant h2 E20 h1 E1 et h2 E200 h1 E1, on obtient les variations relatives de I1 et I2 en fonction des variations
relatives de E20 et E200 :
I1 = o (∆E20 ) + o (∆E200 ) I1 (3.17) 3.4. 3.4.1 Trop peu de données étant disponibles pour déterminer de façon théorique les variations de E20 et E200 au cours de la sorption, les données considérées (voir tableau 3.1) sont déduites de mesures en milieu liquide de toluène à l'aide de dispositifs à ondes acoustiques [Li05] utilisant du PIB comme couche
sensible
.
Tab. 3.1: Modification du module de Young complexe du PIB en fonction de la concentration de toluène Variation Concentration de toluène Concentration dans Variation 3 3 0 dans le liquide (g/m ) la couche (kg/m ) de E2 (%) de E200 (%) 57,6 8,06 -2,08 20,8 115 16,13 -4,17 41,7 172 24,19 -6,25 62,5 230 32,26 -8,33 83,3 288 40,32 -10,42 104,1
Bien que les données proposées dans le cadre des dispositifs à ondes acoustiques semblent très éloignées, notamment en terme de fréquence de sollicitation, de celles requises pour des micropoutres, on supposera que les taux de variations de E20 et E200 sont comparables. Ainsi, on normalise les variations présentées aux valeurs de E20 et E200, en prenant soin de les faire correspondre aux concentrations de toluène dans la couche sensible.
3.4.2 Modification de la fréquence de résonance
À partir de l'expression (3.16) et compte tenu que l'influence de la variation de l'axe neutre peut être négligée, on peut distinguer deux effets lors de la sorption d'un gaz. Le premier est la variation de masse volumique de la couche sensible et le deuxième est la variation de la partie réelle du module de Young complexe de la couche sensible. Ainsi, pour de petites variations de la fréquence de résonance, on peut écrire :
fcouche ∆μ12 I2 ∆E20 =− + fcouche μ12 2 (E1 I1 +
E20 I2 ) (3.19) où ∆f est la variation de fréquence, ∆μ12 est la variation de masse par unité de longueur et ∆E20 et la variation de la partie élastique (E20 ) du module de Young complexe de la couche sensible. On peut ainsi remarquer que, la sorption de toluène augmentant la masse de la couche (∆μ12 > 0) et diminuant l'élasticité de la couche sensible (∆E20 < 0), les deux effets sont cumulatifs en terme de décalage de fréquence. 3.4.3 Modification du facteur de qualité
À partir de l'équation (3.14), on peut déterminer l'influence de la modification de E20 et E200 sur le facteur de qualité associé aux pertes dans la couche sensible. Pour de faibles variations, on peut exprimer la variation relative de ce facteur de qualité par l'expression : ∆E20 E20 Qcouche − 12 ∆Qcouche = Qcouche E20 E200 Q2couche 3 ∆E200 + E200 1 Qcouche − 2 (3.20) Ce qui, dans le cas de faibles pertes, peut s'approximer par : ∆E20 E20 ∆E200 ∆Qcouche = Q + Qcouche couche Qcouche E20 E200 E200 (3.21) Cette expression permet de montrer que, en général, les variations de l'élasticité et des pertes dans la couche sensible peuvent toutes les deux modifier le facteur de qualité. En revanche, dans le cas du PIB détectant du toluène et pour les fréquences de résonance habituelles des micropoutres (1 kHz < f0 < 1 MHz), la partie imaginaire du module de Young complexe est légèrement plus importante que sa partie réelle (E200 > E20 ). De plus, la modification de E200 est aussi plus importante que celle de E20 (∆E200 > ∆E20 ). Ainsi, la variation du facteur de qualité associé aux pertes viscoélastiques est principalement lié à la modification de la partie imaginaire du module de Young complexe. À partir du tableau 3.1, et en extrapolant de façon linéaire pour des plus faibles concentrations, on peut remarquer que les variations de E200 ne sont significatives que pour des concentrations dans la couche sensible au delà de 1 kg/m3 soit 1,7 g/m3 (457 ppm) dans l'air. De plus, la modification du facteur de qualité lié aux pertes viscoélastiques n'influe sur le facteur de 90 qualité total que si les pertes viscoélastiques ne sont pas nég ables devant les autres pertes. La figure 3.13 illustre la modification du facteur de qualité total pour une poutre de 100 μm de long, 30 μm de large, 5 μm d'épais et totalement recouverte de 1,3 μm de PIB. 300
280 260 240 220 200 180 160 140 120 -5 10 -4 10 -3 10 -2 10 -1 10
Concentration de toluène (kg/m3)
Fig.
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1 Substanzialismus und Relationismus
47
(z. B. Roth 1996: 316, 324) sowie in dem Axiom, dass wir in der ›Wirklichkeit‹ Annahmen über
die ›Realität‹ treffen, ohne freilich eine ontologische Entsprechung unterstellen zu dürfen, also
in der Kombination von ontologischem Realismus und erkenntnistheoretischem Idealismus,
derzufolge eine bewusstseinsunabhängige Realität existiert, sie aber prinzipiell nicht erkennbar
bzw. erfahrbar ist (vgl. Roth 1996: 340f., 358),23 finde eine Reformulierung des »Kantschen
Ding-an-sichs als einer nützlichen Prämisse biologischer Kognitionstheorie« (Rustemeyer
1999: 470) statt, die »jedoch ontologisch nicht ernst genommen werden soll. Sie supponiert allerdings
genau das, was ein konsequent gedachtes konstruktivistisches Argument bestreiten müßte:
Realität als das Andere aller Beschreibungen«. (Rustemeyer 1999: 470; Herv. F.S.) In der Tat
erfährt man bei Roth (1996) zwar einiges über epistemologische Hintergrundsannahmen des
neurobiologischen Konstruktivismus, eine Auseinandersetzung mit ontologischen Fragen
sucht man jedoch vergebens. Dies liegt nun m. E. nur zum Teil daran, dass sich ›der‹ Konstruktivismus in erster Linie als eine Erkenntnistheorie, als eine Auseinandersetzung mit dem
Denken versteht (Arnold 2007: 54; ähnlich z. B. Siebert 2000). Denn dann müsste man einfach
nur an einem anderen Ort suchen, um Auseinandersetzungen mit ontologischen Fragen zu
finden. Doch da auch dort – z. B. in Diskursen über kognitivistische Theorien des Lernens –
sucht man vergebens nach der (systematischen) Thematisierung ontologischer Fragen.
Diese Beobachtung legt den Schluss nahe, dass die Zurückhaltung gegenüber ontologischen Fragen andere Ursachen hat als das Desinteresse einzelner Theoretiker. Nach meinem
Dafürhalten sind zwei Ursachen für die Zurückhaltung entscheidend.24 Erstens: Da angenommen wird, ›Ontologie‹ sei, der klassischen aristotelischen Bestimmung folgend, mit dem Sein
selbst befasst und stelle sich – u. a. neben der Kosmologie und philosophischen Theologie –
als ein Themenbereich der Metaphysik (sogenannte ›Erste Philosophie‹) dar, welche als ›Wissenschaft vom Seienden als Seiendem‹ nach den ersten Ursprüngen und Gründen fragt, erscheint es so, als würde derjenige, der ontologische Fragen stellt, Metaphysik betreiben. Das
heißt in anderen Worten: Ontologie zu betreiben, heißt Metaphysik zu betreiben. Weil aber
metaphysisches Denken vom nachmetaphysischen Denken ›verdrängt‹ worden ist, erweist sich
als Tor, wer metaphysische Fragen in Form ontologischer Fragen stellt.25 Zweitens fügt sich
die Zurückhaltung ein in einen, insbesondere von Richard Rorty (1981) diagnostizierten, Entwicklungstrend der neuzeitlichen Philosophie, dessen wichtigstes Kennzeichen ein SichEntfernen »von dem Modell einer Ontologie des Seins und dem entsprechenden ›Zuschauermodell‹ des Wissens, das nach Fundamenten der Erkenntnis strebt« (Reckwitz 2000a: 39)
darstellt. Es liegt also die Vermutung nahe, dass der doppelte Abschied – die philosophische
»Distanzierung vom Projekt einer ›theoretischen‹ Ontologie des Seins« einerseits und von der
»klassischen ›Erkenntnistheorie‹« andererseits (Reckwitz 2000a: 39f.) – dazu geführt hat, dass
man das Kind mit dem Bade ausgeschüttet hat, also sich zwar um eine nachklassische Erkenntnistheorie bemüht hat, darüber jedoch vergessen hat, eine nachklassische Ontologie zu
entwickeln. Mithin stellte die klassische Ontologie eine bloße Negativfolie dar, und es waren
somit nicht die Voraussetzungen für eine Auseinandersetzung mit ontologischen Prämissen
für das Nachdenken über Lernen gegeben.
23
24
25
Vgl. für eine ausführliche Darstellung Abschnitt 1.1.2.2.
Da eine genaue Betrachtung der Ursachen für die Zurückhaltung weit in philosophische Diskurse führen würde
und dies hier nicht geleistet werden kann, belasse ich es dabei, die zwei Ursachen als heuristisch wertvolle Behauptungen und entsprechend plakativ einzuführen.
Vgl. für eine Abriss der beiden Wege, Philosophie zu betreiben Hindrichs (2008), insbesondere dort die Einleitung. Dass auf der Basis eines nachmetaphysischen Verständnisses von ›Ontologie‹ durchaus ontologische Fragen
gestellt werden können, wird in Abschnitt 1.3 dargelegt.
48
Erster Teil: Aufschluss
Der Subjekt-Objekt-Dualismus hat seine ursprüngliche Fassung im sogenannten Cartesianismus. Diesem liegt bekanntlich Descartes' Unterscheidung zwischen (ruhender) ›res cogitans‹, in der Geist, Seele, Vernunft bzw. Verstand beheimatet sind, und ›res extensa‹, also der
u. a. durch Ausdehnung und Bewegung gekennzeichneten äußeren Körperwelt, zugrunde. Als
problematisch werden an der Unterscheidung – auch hiermit ist nichts grundlegend Neues behauptet – zwei Aspekte empfunden. Erstens wird kritisch gesehen, dass der Vorgang des Unterscheidens insofern verselbstständigt hat, als die unterschiedenen ›Dinge‹ (res) als voneinander getrennt gedacht werden.26 Hierauf heben dann Kritiken an dichotomischen UnterUnterscheidungen ab – z. B. die Kritik an den Dichotomien »Sprache versus Handeln, psychische Innenwelt versus soziale Außenwelt«, welche »in die moderne Psychologie eingewobene
Denkmodelle« seien (Kraus 2001: 240). Zweitens werden, mit dem ersten Kritikpunkt eng
verwoben, »die cartesianischen Trennungen zwischen Ich und Welt, Geist und Körper« (Joas
1996: 231) oder die »eindeutige Trennung zwischen innen und außen, zwischen Subjekt und
Objekt, zwischen Lehren und Lernen« (Arnold 2007: 59; ähnlich: Arnold 2007: 82f.; Arnold/Siebert 2006: 4f.) oder der Dualismus zwischen einer Innenwelt des Bewusstseins und
einer (sozialen) Außenwelt bzw. zwischen ›Subjekt‹ und ›Objekt‹ oder zwischen ›Individuum‹
und ›Gesellschaft‹ (vgl. Reckwitz 1997b: 318, 322) kritisch gesehen.27 Unter dem Label ›Cartesianismus‹ bzw. ›cartesianisch‹ ist also eine Reihe von (Teil-)Problemen subsumierbar. Sie
lassen sich anhand von drei – in der Leitunterscheidung einer Innen-Außen-Differenz aufeinander verweisenden und in die ontologische und erkenntnistheoretische Dimension des Subjekt-Objekt-Dualismus eingehenden – Fragen grob ordnen: der Frage nach dem Zusammenhang oder Verhältnis von Körper und Geist, der Frage nach dem Zusammenhang oder Verhältnis von Individuum und Gesellschaft, der Frage nach dem Zusammenhang oder Verhältnis
von erkennendem Subjekt und erkanntem Objekt.
In jüngerer Zeit wurde das ›Körper-Geist-Problem‹ u. a. im Umkreis der analytischen Philosophie des Geistes und in den Diskursen rund um die Frage nach der ›Naturalisierung des
Geistes‹ als zum ›psychophysischen Problem‹ oder ›Geist-Gehirn-Problem‹ transformiertes
›Leib-Seele-Problem‹ diskutiert. Die Einwände gegen die »cartesianische Dichotomie von Körper und Geist« (Schneider 1998: 291) bzw. den Dualismus von Körper und Geist sind vielfältig und werden hier allenfalls am Rande miteinbezogen.28 Bekannt sind Gilbert Ryles Kritik an
Mythos bzw. »Dogma vom Gespenst in der Maschine« (Ryle 1969: 13) und Antonio Damasios
Vorgehen gegen »Descartes' Irrtum«, der v. a. in der »abgrundtiefen Trennung von Körper
und Geist«, in der »Trennung der höchsten geistigen Tätigkeiten vom Aufbau und der Ar26
27
28
Dies zieht dann u. a. die Diagnose nach sich, dass cartesianisches Denken ein »Denken in Ursachen und Wirkungen« (Arnold 2007: 80) ist.
Es wird an den Zitaten und im Weiteren deutlich, dass die Termini ›Dualismus‹ und ›Dichotomie‹ nahezu synonym gebraucht werden können. Sie bezeichnen den Umstand, dass zunächst eine Grenze oder Trennwand oder
desgleichen gezogen wird; von hier aus geht man dann davon aus, dass die auseinandergeschnittenen Teile als ›an
und für sich‹ stehend behandelt werden können. Dagegen wird bei einer ›Dualität‹ (Giddens) zwar auch unterschieden, jedoch ist das Ergebnis der Unterscheidung mehr eine Linie, die auf die ›Einheit‹ gelegt wird und die die
Bezogenheit der ›Teile‹ aufeinander nicht negiert, sondern eine Berührungslinie darstellt. Vgl. Abschnitt 1.4 zur
Verdrehung eines Distanzierungs- oder Unterscheidungsaktes in eine Grenze oder Scheidewand im Rahmen des
Modells des ›homo clausus‹. Zum ›Zwischen‹ als einer »Sphäre der Differenzierung« (Waldenfels 2000: 286) vgl.
Abschnitt 7.1.1.
Im Weiteren dürfte deutlich werden, dass diese Diskurslinien auch für das Nachdenken über Lernen von Relevanz sind, beispielsweise für die Kritik an (bestimmten) kognitivistischen Lernkonzepten (vgl. Abschnitt 2.3.3)
oder für die möglichen Aporien physikalistischer bzw. naturalistischer Erklärungsansätze, wobei hier insbesondere dem auf einer Substanzialisierung bzw. Verdinglichung aufbauenden Kategorienfehler der »Hypostasierung
theoretischer Konstrukte« (Hartmann 2006; vgl. Abschnitt 1.4) besondere Aufmerksamkeit geschenkt wird.
1 Substanzialismus und Relationismus
49
beitsweise des biologischen Organismus« bestand (Damasio 2004: 330; vgl. 328–333) und die
Annahme enthält, dass es »zwei verschiedene Arten von Existenz oder Sein gibt« (Ryle
1969: 9). Von besonderem Interesse für die vorliegende Studie ist Maurice Merleau-Pontys
antidualistischer Ansatz. In diesem Ansatz wird ein Dualismus von Körper und Geist unterlaufen. Denn: »Konkret genommen, ist der Mensch nicht ein Psychismus, verbunden mit
einem Organismus, sondern das Kommen und Gehen der Existenz, die bald sich körperlich
sein läßt, dann wieder in persönlichem Handeln sich zuträgt.« (Merleau-Ponty 1966: 113) An
die Stelle der Frage, ob denn der Körper oder der Geist der Ausgangs- und Referenzpunkt der
Analyse sein solle, tritt in Merleau-Pontys Phänomenologie, wie auch in deren Rezeption (z. B.
Joas 1996, Meyer-Drawe 1987, Coenen 1985, Waldenfels 2000), die Frage nach dem ›Leib‹
bzw. der ›Leiblichkeit‹ bzw. dem ›Leibkörper‹ in den Vordergrund. Die Frage nach dem Leib
wird in unserem Zusammenhang ›ontologisch zugespitzt‹, da der »menschliche Leibkörper als
Ort des Sich-Verklammerns verschiedener Schichten (anorganische, organische, seelische,
geistige), damit als ontologisch dichtester Ort im Kosmos entdeckt« (Fischer 2006: 327; Herv. F.S.)
werden kann und »so als Konstitutionszentrum der menschlichen Sphäre« (Fischer 2006: 327)
rekonstruierbar ist.29
Ferner ist die cartesianische Trennung in Form einer »Absperrung individueller Affektimpulse von der motorischen Apparatur, von der unmittelbaren Steuerung der Körperbewegungen, von Handlungen« (Elias 1997a: 68) als Ursache für die »Vorstellung von den absolut unabhängig voneinander entscheidenden, agierenden und ›existierenden‹ Einzelmenschen« (Elias
1997a: 68) verstehbar. Das »Muster der Innen-Außen-Differenz« (Reckwitz 1997b) offenbart
sich hier an der sozialtheoretischen Frage nach der Konzeptualisierung und dem Zusammenhang bzw. dem Verhältnis von ›Individuum‹ und ›Gesellschaft‹ und mittelbar an der methodologischen bzw. wissenschaftstheoretischen Frage nach dem geeigneten Ausgangspunkt für den
Zugang zum Subjekt-Objekt-Dualismus, expliziert an der Frage nach der Überwindung des
Mikro-Makro-Problems bzw. des Problems von Struktur und Handeln.30
Ontologisch zugespitzt prominiert die Frage nach der Konzeptualisierung und dem Zusammenhang bzw. dem Verhältnis von ›Individuum‹ und ›Gesellschaft‹ als Frage nach den
Konzeptualisierungen und möglichen Verhältnissen von ›Person‹ und ›Welt‹. In Descartes'
Subjektphilosophie zeigt sich dabei ein ontologischer Dualismus: hier die introspektiv zugängliche, private ›res cogitans‹ (Innenwelt, Bewusstsein), dort die ›res extensa‹, die von der (Natur-)Wissenschaft erforschte und erforschbare ausgedehnte Objektewelt (vgl. z. B. Reckwitz
1997b: 322). Entsprechend ist beispielsweise Ryles Cartesianismus-Kritik – Ryle kritisiert die
cartesianische »Zweiweltenlegende« (Ryle 1969) – auch ontologisch verstehbar. Gleiches kann
für die Leitdifferenz von ›inneren‹ subjektiven Wünschen und ›äußeren‹ sozialen Erwartungen
bzw. sozialem Zwang (vgl. z. B. Reckwitz 2000a: 125f.) und für das »cartesianische Menschenbild des geschlossenen Subjekts« (Coenen 1985: 72) gelten. Auch die Kritik an einem dualisti29
30
Dies heißt freilich nicht, dass damit schon der Körper-Geist-Dualismus gänzlich überwunden ist. Vielmehr ist
damit ein anderer begrifflicher Anfang für menschliches Lernen gesetzt, dessen Implikationen noch bedacht werden müssen. Man vergleiche hierzu etwa Bernhard Waldenfels' Einschätzung: »In der Interpretation des leiblichen
Verhaltens haben wir, so mag es scheinen, den Dualismus von Geist und Natur, von Innen und Außen scheinbar
für immer hinter uns gelassen. […] Und doch, eines sollte uns stutzig machen. Zum Beispiel fordert Giddens (1997: 358): »wir müssen den Dualismus von Objektivismus und Subjektivismus
überwinden«.
50
Erster Teil: Aufschluss
schen Lern- und Bildungsbegriff (z. B. Holzapfel 2002: 184) oder an dualistischen Lerntheorien (z. B. Lave 1997a, Lave/Wenger 1991) kann als Kritik an einem ontologischen Dualismus
verstanden werden, so etwa, wenn darauf insistiert wird, dass in konventionellen Lernkonzepten
das Subjekt-Welt-Verhältnis dualistisch angelegt sei, weil Subjekt und Welt als zwei unterschiedliche, voneinander getrennte Entitäten gefasst seien, die einander nicht definierten (vgl.
Lave 1997a: 122), weil Wissen als im Gehirn befindlich angesehen werde und das Lernen als
Transmission und Assimilation von Äußerem, kulturell Vorgegebenem gefasst sei (vgl. Lave/Wenger 1991: 47).
Allerdings wird eine »ontology of two realms« (Packer/Goicoechea 2000: 228) regelmäßig
nicht explizit thematisiert. Daher liegt der Schluss nahe, dass die Kritik an dualistischen Lernkonzepten mehr an der Frage nach dem Zusammenhang oder Verhältnis von erkennendem
Subjekt und erkanntem Objekt ausgerichtet ist.
Geradezu prototypisch ist die bereits erwähnte ›konstruktivistische‹ Kritik an der tradierten Innen-Außen-Differenz von erkennendem Subjekt und ihm gegenüberstehender Außenwelt (z. B. Arnold 2005), die sich als Kritik an einem erkenntnistheoretischen Realismus versteht (vgl. z. B. Roth 1996, Arnold 2005: 40, Arnold 2007: 54) und postuliert, dass Erkennen
von operational geschlossenen kognitiven und emotionalen »Systemen« geleistet wird (vgl.
z. B. Arnold 2005) und dass der Objektbegriff im erkennenden Subjekt aufgelöst wird (vgl.
Arnold 2007: 83, Arnold/Siebert 2006: 4f.). Arnold und Siebert (2006: 4f.) beklagen den Umstand, dass sie sich nicht mit »Neorealisten« verständigen könnten, weil ihre eigenen Sprachmittel noch nicht ausreichten. Weiter ließen sie sich von »Neorealisten« und der Frage nach
der Subjekt-Objekt-Vermittlung einschüchtern. Nach meinem Dafürhalten bleibt die Pattsituation zwischen ›(Neo-)Realismus‹ und ›Konstruktivismus‹ solange erhalten, wie nicht deutlich
gemacht wird, dass sie erfolgreich aneinander vorbeireden, etwa wenn Realisten am Konstruktivismus nicht dessen erkenntnistheoretischen Idealismus, sondern dessen ontologischen Realismus kritisieren, und wenn Konstruktivisten einen erkenntnistheoretischen Realismus kritisieren, ontologischen Fragestellungen hingegen regelmäßig ausweichen (vgl. Mitterer
1999: 486). Insofern erscheint eine Bedingung der Möglichkeit das erwähnte Aneinandervorbei-Reden beenden zu können in der Reflexion über ontologische Fragestellungen zu bestehen – vorausgesetzt, das gemeinsame Ziel besteht darin, den erkenntnistheoretischen und
den ontologischen Subjekt-Objekt-Dualismus, die dualistische Denkweise selbst (vgl. Mitterer
1999), zu überwinden. Im Speziellen beruht nach meinem Dafürhalten Arnolds und Sieberts
Erklärungsnot gerade darauf, dass die Rede von der Auflösung des Objektbegriffs im erkennenden Subjekt (Arnold 2007: 83, Arnold/Siebert 2006: 4f.) ihrem Anspruch nach einen nichtdualistischen Begriff des Erkennens setzt, es aber versäumt, der alternativen Erkenntnistheorie
›ontologischen Halt‹ zu verschaffen, d. h., eine nicht-dualistische Ontologie bzw. eine ›passende‹ alternative Subjektkonzeption, eine ›passende‹ alternative Weltkonzeption und eine ›passende‹ alternative Konzeption des Subjekt-Welt-Verhältnisses beizustellen.
An dieser Stelle ist jedoch sogleich Eingrenzungsarbeit vonnöten. 1. Wenn es stimmt, dass
Erkennen bzw. (wissenschaftliches) Beschreiben unvermeidlich dualistisch ist, d. h., die Trennung zwischen Erkenntnissubjekt und Erkenntnisobjekt für wissenschaftliches Erkennen unhintergehbar ist (Laucken 2003: 32–40, 46, 305), dann ist ›nicht-dualistisches‹ wissenschaftliches Erkennen – um ein Wort von Wittgenstein abzuwandeln (»Philosophie dürfte man
eigentlich nur dichten«; Wittgenstein 1994: 483; zit. n. Bezzel 2000: 95) – nur als Dichtung
möglich. Oder als ›leibliches Erkennen‹ – sofern die Prämisse gelten soll, dass wissenschaftliches Erkennen als menschliche Tätigkeit nicht leibentbunden erfolgt. Oder in anderer Weise.
Kurz gesagt: Die alternative Erkenntnistheorie wird in dieser Untersuchung als ein Desiderat
1 Substanzialismus und Relationismus
51
geführt. 2. Ich sehe es nicht als meine Aufgabe an, den Konstruktivismus, wie er von Arnold
und Siebert vertreten wird (z. B. Arnold/Siebert 2005, 2006, Siebert 2000, Arnold 2007), mit
›ontologischem Halt‹ zu versorgen. Dies aus dem einfachen Grund, dass dies m. E. nicht möglich ist, da durch die Anleihen beim neurobiologischen Konstruktivismus (Roth, Maturana) das
›lernende Subjekt‹ letztlich nach dem Bild des ›homo clausus‹ (Elias) gebaut ist und da die
operational geschlossenen kognitiven und emotionalen ›Systeme‹, bildlich gesprochen, in dem
geschlossenen Subjekt (vgl. Coenen 1985: 72) eingeschlossen sind. Dies (nicht nur für ›den‹
Konstruktivismus) aufzuzeigen, wird eines der Hauptanliegen der weiteren Ausführungen sein.
1.1.2
Zwei Stränge der Kritik an der klassischen Erkenntnistheorie und dem zugehörigen Modell des
›homo philosophicus‹
Die ›klassische‹ Erkenntnistheorie befasst sich besonders mit dem Verhältnis von erkennendem Subjekt und erkanntem Objekt sowie mit dem Problem der ›Außenwelt‹ (IdealismusRealismus-Problem) (Edelmann 2000: 115). Sie arbeitet mit der zentralen Annahme, dass ein
Wissen erlangt werden kann, welches »als notwendig, ewig und universell gelten kann«
(Göhlich/Zirfas 2007: 35). Bekanntlich stehen sich in der Frage, wie dieses Wissen erlangt
werden kann, Empirismus und Rationalismus gegenüber. Während der Empirismus den Ausgang der Erkenntnis bei den Empfindungen, der Erfahrung, nimmt und die ›Realität‹ induktiv
durch richtigen, ordnenden Vernunftgebrauch erschließt, nimmt der Rationalismus den deduktiven Weg von der Vernunft, den reinen Prinzipien des Denkens, ausgehend die logische Ordnung der ›Realität‹ zu erfassen (vgl. hierzu etwa Göhlich/Zirfas 2007: 36–39).31 Es würde den
Rahmen der vorliegenden Arbeit sprengen, allen Verästelungen der Kritik an der klassischen
Erkenntnistheorie nachzugehen. Ich begnüge mich daher mit einer Zusammenstellung der
wesentlichen Kritikpunkte. Die (jüngere) kritische Auseinandersetzung mit der klassischen
Erkenntnistheorie bezieht sich einerseits eher allgemein auf die klassische Erkenntnistheorie
(z. T. inklusive der Kritik an einem Rationalismus bzw. Empirismus) und speziell auf das darin
enthaltene Bild vom erkennenden Subjekt, andererseits auf das Idealismus-Realismus-Problem.
1.1.2.1 Grundzüge einer allgemeinen Kritik an der klassischen Erkenntnistheorie sowie einer
konkreten Kritik am Modell des ›homo philosophicus‹
Der erste Strang der Kritik an der klassischen Erkenntnistheorie besteht in einer ›allgemeinen‹
Kritik und in der Kritik am Erkenntnissubjekt dieser Erkenntnistheorie. Sie hängen miteinander zusammen und finden in der Auseinandersetzung mit der ›Krise der Repräsentation‹ einen
wesentlichen Konvergenzpunkt.
Die Ansätze der Wissenschaftler und Wissenschaftlerinnen, welche eine Kritik an der klassischen Erkenntnistheorie formuliert haben, lassen sich sicherlich nicht in ihrer Gänze unter
ein ›Paradigma‹ bringen. Dies ist auch gar nicht notwendig, um, bei allen Unterschieden, die
Gemeinsamkeit der gängigen Kritik an der klassischen Erkenntnistheorie zu beschreiben. Mit
dem Term ›Krise der Repräsentation‹ (vgl. z. B. Reckwitz 2003b) ist die Gemeinsamkeit tref31
Da in diesem Abschnitt die Frage nach dem erkenntnistheoretischen Subjekt-Objekt-Dualismus im Vordergrund
steht, ist die Frage, ob dem induktiven oder dem deduktiven Weg der Vorzug zu geben sei, nicht von Interesse.
Nur so viel: Das Empirismus wie Rationalismus zugrunde liegende ›Problem‹ besteht in der impliziten Konzeption des Menschen, welche Norbert Elias als ›homo philosophicus‹ umschrieben hat: Für Elias steuert auf der Basis des Bildes des ›homo philosophicus‹ das Denken »hilflos zwischen der Scylla irgendeines Positivismus und der
Charybdis irgendeines Apriorismus hin und her« (Elias 1997a: 51).
52
Erster Teil: Aufschluss
fend umrissen. Die Kernaussage der Behauptung einer Krise der Repräsentation, welche im
Kontext des sogenannten Postempirismus eine fast schon kanonisch zu nennende Ausarbeitung erfahren hat und epistemologisch wie auch wissenschaftstheoretisch gemeint ist (vgl.
Reckwitz 2003b), besteht in der »Kritik an Korrespondenz- und Abbildtheorien« (Reckwitz
2000a: 22) bzw. der Kritik an einer »Abbildtheorie der Wahrheit« (Zielke 2004: 142) und einer
»Korrespondenztheorie der Wahrheit« (Zielke 2004: 253):
Als Ergebnis dieser epistemologischen Überlegungen erscheint es nicht länger begründbar, eine […]»Korrespondenz« zwischen wissenschaftlichen Theorien und einer unabhängig davon zu denkenden Welt der »Tatsachen«, hier der sozialen Tatsachen, anzunehmen: Wissenschaftliche Theorien registrieren nicht unabhängig von
ihnen selbst vorfindbare Bedeutungen der Welt, sie produzieren erst die Bedeutungen auf kontingente Weise.
(Reckwitz 2000a: 23)
Vor allem Richard Rorty (1981, 1989) übte Kritik an dem in der westlichen intellektuellen
Tradition verankerten erkenntnistheoretischen Projekt der Gewinnung von ›Wahrheit‹. Es ist,
so Rorty, gescheitert (vgl. Reckwitz 2003b). Rorty verwirft im Anschluss an Heidegger, Dewey32 und Wittgenstein33 den »Fundamentalanspruch der überkommenen Erkenntnistheorie«
(Schneider 1998: 156). In ähnlicher Weise ging mit Putnams Distanzierung von einem metaphysischen Realismus, eine Distanzierung von einer »Korrespondenztheorie der Bezugnahme«, einer Theorie, die eine »Ähnlichkeit zwischen Repräsentation und äußerem Ding«
(Schneider 1998: 157) annimmt, einher (vgl. Putnam 1981, 1999).34
Vor dem Hintergrund eines differenztheoretischen Ansatzes stellt Rustemeyer die Kritik
am klassischen Wissens- und Erkenntnismodell dar. Die Kritik stelle drei Prämissen des Repräsentationsmodells des Wissens infrage, nämlich die Prämissen, dass Unterschiede und nicht
Unterscheidungen repräsentiert sind, dass die Unterschiede intelligibel sind und dass sie zeitlos
sind. Die Infragestellung geschieht dabei auf der Basis eines nachklassischen Wissensmodells,
das an den Unterscheidungen ansetzt, welche bereits in der Wahrnehmung fundiert sind und
eine kontextuelle Varianz zulassen, sodass sich Wissen als ein Prozess dynamischer Ordnungsbildungen darstellt (Rustemeyer 2005: 15).
32
33
34
Für eine Einführung in die Aktualität von Deweys Absage an letzte Gewissheiten und Wahrheiten im Zuge der
Auseinandersetzung mit der klassisch-abendländischen Metaphysik und Epistemologie vgl. z. B. Bauer (2005).
Den Bezug zwischen Deweys Kritik an der klassischen Erkenntnistheorie zu lerntheoretischen Fragen stellt z. B.
Faulstich (2005a: 532ff.) her.
Zu Wittgensteins Kritik des neuzeitlichen Mentalismus, insbesondere seiner Kritik der Repräsentationsauffassung
der Sprache vgl. z. B. Cursio (2006: 55–72). Für eine Einführung in Wittgensteins Denken allgemein vgl. z. B.
Skirbekk/Gilje (1993b: 848–854) und Bezzel (2000).
Die Absage an eine Korrespondenztheorie der Wahrheit erfolgt auch in Kenneth Gergens Sozialkonstruktionismus. Jedoch mündet nach Zielkes Auffassung Gergens antirealistische Auffassung von Erkenntnis und seine Kritik an einer Abbildtheorie der Sprache oder einer repräsentationalistischen Sprachauffassung in einen »anonymen
Praxisbegriff« (Zielke 2004: 299) ein (vgl. Gergen 1994a, 1994b, 1997, 1999, Zielke 2004: 201–300; vgl. auch Abschnitt 7.1.1). Dies habe zur Folge, dass bei Gergen Theorien von ihrer empirischen Grundlage abgekoppelt werden, d. h.: Gergen sage sich faktisch von der Psychologie als empirischer Wissenschaft los, der Psychologie gehe
ihr Gegenstandsbereich (der Akteur) verloren, sie werde auf reine Text- oder Sprachwissenschaft reduziert (Zielke
2004: 228). Für eine Kritik an der Wissenschaftsphilosophie des Sozialkonstruktionismus von Gergen vgl. etwa
Ratner (2005). Ratner kritisiert u. a., dass im Sozialkonstruktionismus Wahrheitsansprüche ausgeschaltet würden
und dass aus dem Sozialkonstruktionismus unkritisches Denken hervorgehe. Eine Replik findet sich etwa bei
Zielke (2006b), wobei der Schwerpunkt darauf liegt, erstens den Antirealismus Gergens als gegen bestimmte Varianten des Realismus (nämlich repräsentationistische Vorstellungen) zu spezifizieren und zweitens die Grenzen
des Sozialkonstruktionismus (insbesondere seinen »anonymen Praxisbegriff«; Zielke 2004: 299) darzustellen und
den Ausblick auf eine pragmatistisch-kulturpsychologische Handlungstheorie zu eröffnen (letzteres ähnlich schon
in Ansätzen bei Zielke 2004, systematischer bei Zielke 2006a). Ein Kommentar zu der Zielke-Ratner-Diskussion
erfolgte durch van Oorschot/Allolio-Näcke (2006).
1 Substanzialismus und Relationismus
53
Norbert Elias, schließlich, richtet einen Teil seiner Kritik an der klassischen philosophischen Wissenschafts- und Erkenntnistheorie an deren statischen Charakter aus. Er hält fest,
dass es üblich sei, »sich vorzustellen, daß das Wissen zwar veränderlich sei und wachsen könne, daß aber die Denktätigkeit des Menschen selbst ewigen und unveränderlichen Gesetzen
unterliege« (Elias 2004: 42).
Aber diese gedankliche Trennung einer ewigen Form des Denkens von seinen wechselnden Gehalten beruht nicht auf
einer Untersuchung der Sachverhalte selbst, sondern liegt in dem menschlichen Sicherheitsbedürfnis begründet,
hinter allem Wandelbaren das absolut Unwandelbare zu entdecken. (Elias 2004: 42; Herv. i. Orig.)
Man akzeptiere, dass das Unwandelbare hinter allem Wandel »einen höheren Wert besitzt als
der Wandel selbst« (Elias 2004: 42; entfernt ähnlich: Waldenfels 2000: 123). Der Rekurs auf
Unwandelbares (z. B. ›Logik‹, ›Kategorien‹, Gesetzesmäßigkeiten des Denkens) beim Nachdenken über das Sich-Wandelnde (z. B. gesellschaftliche Prozesse) beruhe auf einer Verwechslung von Tatsache und Ideal, weshalb Elias eine Art sachbezogene, deskriptive Wissenschaftsund Erkenntnistheorie fordert (vgl. Elias 2004: 42f.).
Der zweite Punkt ist eine direkte Kritik am Erkenntnissubjekt der klassischen Erkenntnistheorie. Sie wird im postempiristischen Denken als Infragestellung der Denkfigur des »reinen, ›unbeobachteten Beobachters‹« (Reckwitz 2004b: 14) und als Abschied vom »›Zuschauermodell‹ des Wissens, das nach Fundamenten der Erkenntnis strebt« (Reckwitz 2000a: 39) formuliert und bezeichnet das, was Elias als Distanziertheit des Denkenden von den ›Erkenntnisgegenständen‹ umschrieben hat (vgl. Elias 1997a: 63).
Der Denkende bzw. Erkennende ist in der (alt-)europäischen Geist-Philosophie das ›monologische Subjekt‹ oder das reine monologische Bewusstsein (vgl. v. a. Taylor 1993: 49; ähnlich: Keupp 2001: 50). Es ist monologisierend, weil es in einem inneren Raum, unabhängig
vom Körper oder von anderen operiert.
Die Kritik am ›monologischen Subjekt‹ wird auch von konstruktivistischer Seite her formuliert, beispielsweise in der Absage an die Auffassung, die Welt sei eine Konstruktion35 eines
›monologischen Subjekts‹ (z. B. Arnold/Siebert 2005: 59f.), oder in der Ablehnung eines einsamen, monologischen Ichs, das vor jeder Bezugnahme auf ›Welt‹ existiert (Maturana/Varela
1987: 254).36 Allerdings verhaken sich jene Konstruktivismen, die an naturalistischen Vokabularen Halt suchen – wie bereits angedeutet wurde und wie weiter unten noch genauer zu zeigen sein wird – im Dualismus zwischen einer physischen oder geistigen Welt.
Eine, m. W. fast in Vergessenheit geratene, Kritik am ›monologischen Subjekt‹ stammt
von Elias. Sie stellt den zweiten Teil seiner Kritik an der klassischen philosophischen Erkenntnistheorie (und Wissenschaftstheorie) dar. Elias bezeichnet das erkenntnistheoretische Subjekt
der klassischen europäischen Philosophie als »homo philosophicus« (Elias 1997a: 50). Dieses
Menschenbild des ›homo philosophicus‹ ist nach Elias eine der Abarten des ›homo clausus‹37
(Elias 1997a: 53). Der ›homo philosophicus‹ ist als Erwachsener gedacht:
35
36
37
Ohne konstruktivistischen Bezug und ähnlich wie Keupp (2001) legt Zielke als ein Kriterium für einen postkognitivistischen Wissensbegriff Wissen als Konstruktion, also als Leistung eines (körperlichen bzw. leiblichen) Akteurs
bzw. als gemeinsame Leistung mehrerer Akteure fest und widerspricht so dem Repräsentationsmodell (Zielke
2004: 147).
Maturana/Varela sprechen sich daher für eine »fortwährende Transformation im Werden der sprachlichen Welt,
die wir zusammen mit anderen menschlichen Wesen erschaffen« (Maturana/Varela 1987: 254), aus.
Weiter unten werde ich den ›homo clausus‹ als eine Art ontologisches Äquivalent zum ›homo philosophicus‹
ausführlich beschreiben. Hier kommt es lediglich darauf an, auf die dezidierte Unterscheidung zwischen erkenntnistheoretischer Ebene und ontologischer Ebene hinzuweisen, die bei Elias angelegt ist.
54
Erster Teil: Aufschluss
Der einzelne Mensch macht als Erwachsener die Augen auf und erkennt hier und jetzt ganz aus eigener Kraft
und ohne von anderen zu lernen, nicht allein, was alle diese Objekte sind, die er wahrnimmt, […], er erkennt
überdies auch ganz unmittelbar und hier und jetzt, daß sie kausal und naturgesetzlich miteinander verknüpft
sind. (Elias 1997a: 50)
Die kausale Verknüpfung sucht der ›homo philosophicus‹, »der nie ein Kind war und gleichsam als Erwachsener auf die Welt kam« (Elias 1997a: 51), in den Tatsachen ›außerhalb seiner‹
oder in seinem ›Inneren‹ (vgl. Elias 1997a: 51). Mittels dieser Konzeption wird der traditionelle
philosophische Problemansatz »egozentrisch, weil er sich auf die Frage beschränkt, wie ein
einzelner Mensch wissenschaftliche Erkenntnisse gewinnt« (Elias 2004: 43).38
Eine Andeutung davon, dass das Bild des ›homo philosophicus‹ erlernt bzw. über Sozialisationsprozesse erworben wird und insofern ›gesellschaftlich vermittelt‹ ist, bietet eine andere
Textstelle bei Elias: Der Einzelne hat »bestimmte ›Formen des Denkens‹, spezifische Kategorien, besondere Arten, Einzelwahrnehmungen in Zusammenhang miteinander zu bringen,
erworben« (Elias 2004: 43f.). Dies wiederum bedeutet in Elias' Diktion, dass das Bild des ›homo philosophicus‹ ebenso eine »spezifische Selbsterfahrung« (Elias 2004: 128) des (modernen,
westlichen) Menschen ist, wie dies der ›homo clausus‹ ist. Ich werde darauf beim ›homo clausus‹ zurückkommen. Einstweilen ist von Bedeutung: Elias hält dem Bild des ›homo philosophicus‹ (und dem Bild des ›homo clausus‹) entgegen, dass die einzelnen ›von klein auf in
Interdependenz mit anderen leben‹ und doch relativ autonom sind und ›miteinander wandelbare Figurationen bilden‹ – ein Umstand, der mit der Reduktion auf einzelne Erkenntnisakte
eines abgeschlossenen Einzelmenschen nur schwerlich erfasst werden kann (vgl. Elias
1997a: 51).
Die Tragweite dieser scheinbar selbstverständlichen Behauptungen ist nicht zu unterschätzen. Es kommt nämlich die Frage auf, wie die genannte Interdependenz gedacht ist – mithin:
wie Sozialität gedacht ist.39
1.1.2.2 Kritik an der klassischen Erkenntnistheorie ausgehend vom Idealismus-RealismusProblem – Anmerkungen zu den Grenzen eines Festhaltens an einem ontologischen
Realismus
Der zweite Strang der Kritik an der klassischen Erkenntnistheorie befasst sich mit dem Idealismus-Realismus-Problem. Sie wird u. a. in Varianten eines neurobiologischen Konstruktivismus (z. B. Roth 1996) formuliert bzw. in Varianten konstruktivistischer Theoriebildung, die
Anleihen beim neurobiologischen Konstruktivismus nehmen (z. B. Arnold 2007). Ich versuche
in diesem Abschnitt v. a. zu zeigen, dass der Abschied von einem Repräsentationsmodell des
Wissens und einem damit verknüpften Abschied von einer dualistischen Erkenntnistheorie
unvollständig vollzogen wird, solange der Abschied vor dem Hintergrund des Festhaltens an
einem ontologischen Realismus stattfinden soll. Insofern lautet der Appell, entweder eben das
ernst zu nehmen, was Rustemeyer als »das Andere aller Beschreibungen« (Rustemeyer
38
39
Inwiefern Elias' Kritik auf Kant gemünzt ist (vgl. hierzu z. B. Skirbekk/Gilje (1993b: 511): Kant arbeite »mit einer
Erkenntnistheorie, die im Einzelmenschen begründet ist«) und sich mehr an (links-)hegelianischem Denken anlehnt (vgl. hier etwa Skirbekk/Gilje (1993b: 590): Hegels Denken richte sich gegen die individualistische Lehre einer Selbstdefinition des Individuums, gegen Auffassung vom Individuum als in sich selbst ruhend), wäre sicherlich eine interessante Frage, der aber im Weiteren keine Beachtung geschenkt wird.
Auf den Grundunterschied zwischen einem substanzialistischen und damit letztlich dualistischen ›Billardkugelmodell‹ und einem relationalen Modell werde ich in Abschnitt 1.2 eingehen. Elias' Vorschlag, vom Bild des letztlich
isolierten Einzelmenschen zu einem relationalen Bild der ›homines aperti‹ zu wechseln, werde ich in Abschnitt 4.3
aufgreifen. Eine Radikalisierung in Richtung auf eine synchrone Betrachtungsweise findet sich in Abschnitt 5.3.1.
1 Substanzialismus und Relationismus
55
1999: 470) bezeichnet hat – nämlich die ›Realität‹ – oder sich von dem ontologischen Realismus zu verabschieden und sich den in den eigenen Vokabularen verwendeten »Denkmitteln«
(Elias; z. B. Elias 2003: 89, Elias 2004: 74) oder »Denkinstrumenten« (Elias; z. B. Elias
2003: 54) zuzuwenden. Hier liegt im Wesentlichen die Differenz zwischen der konstruktivistischen und der postempiristischen Ablösung vom Repräsentationsmodell des Wissens: Das
postempiristische – und einen Schritt weiter: das poststrukturalistische – Denken ermöglicht
die Revisibilisierung der ontologischen Hintergrundsannahmen, welche in die gegenstandskonstitutiven Vokabulare eingelassen sind, und erlaubt es somit, zwischen substanzialistischen
und relationalen ›Gegenstandsauffassungen‹ zu unterscheiden.40 Dem neurobiologisch fundierten Konstruktivismus gelingt dies hingegen nicht.
Es geht mir ferner darum, einen Eindruck davon zu vermitteln, dass der Abschied vom
ontologischen Realismus (und die Hinwendung zu den Vokabularen) ein notwendiger (Zwischen-)Schritt zur Überwindung von »Zweiweltenlegenden« (Ryle) – sowohl in Form eines
Dualismus zwischen einer physischen und einer geistigen Welt als auch in Form des sozialtheoretischen Dualismus – ist und dass dieser Weg über die grundlegendere Überwindung einer
»Ontologie der Gegenständlichkeit« (Rombach 2003: 17) führen kann.
Hierfür ist eine Konkretisierung notwendig. Der ontologische Realismus, von dem man
sich verabschieden sollte, baut auf dem ›klassischen‹ Verständnis von ›Ontologie‹ im Sinne von
Metaphysik, auf einer »Ontologie des Seins« (Reckwitz 2000a: 39; Herv. F.S.) auf und ist per se
ein Bestandteil dualistischen Denkens. Versteht man hingegen unter Ontologie eine nachmetaphysische Lehre von den »Seinsweisen« (Rombach 2003: 102; Herv. F.S.),41 ist es gar nicht mehr
notwendig, wie etwa Roth eine ›transphänomenale Realität‹ anzunehmen – selbst nicht im
Sinne einer plausiblen Annahme. Vielmehr kann man z. B. annehmen, dass Systeme ›real‹ sind
(Luhmann; vgl. Reese-Schäfer 2001: 37–40), denn sie sind es in dem Sinne, dass sie innerhalb
eines Vokabulars ›existieren‹ – in luhmannscher Diktion: durch Beobachtung hervorgebracht
werden (Krause 2001: 16, 208). Dies hat den großen Vorteil, dass man keinen Bezug mehr auf
metaphysische Konstrukte nehmen muss – auch nicht durch eine bloße Negation, welche vom
Negierten letztlich abhängig bleibt und das Negierte nachgerade voraussetzen muss. Folglich
wird man sich der Sprachspielabhängigkeit der eigenen Konstrukte oder »Denkmittel« (Elias;
z. B. Elias 2003: 89, Elias 2004: 74) bewusster bzw. in die Lage versetzt, ihrer bewusster zu
werden.42
Hierin scheint mir im Übrigen ein Ausweg für einen Konstruktivismus arnoldscher Prägung zu liegen: Wenn Arnold die Wendung des Konstruktivismus zu einer »Beobachtertheorie« beschreibt (z. B. Arnold 2003: 52), so sieht er durchaus das beschriebene Erfordernis.
Aber diese Wendung müsste – und das bleibt m. E. unerkannt – zunächst die Gleichsetzung
von biologischen Systemen bzw. Neuronensystemen und ›Sinnsystemen‹ rückgängig machen.
Dann wäre es möglich, Lernen in seiner »Systemik« (Arnold) zu beschreiben, ohne zwischen
einer inneren und einer äußeren Systemik unterscheiden und sie dann nachträglich aufeinander
beziehen zu müssen.43 In anderen Worten: Für eine Selbstbeobachtung ›des‹ Konstruktivismus
ist es nicht nur nicht notwendig, sondern sogar kontraproduktiv, sich an die neurobiologische
Sichtweise anzuschließen. Denn es handelt sich um zwei unterschiedliche Ontologien, welche
40
41
42
43
Vgl. hierzu Abschnitt 1.2.
Vgl. hierzu Abschnitt 1.3.
Vgl. hierzu z. B. Ritsert (1998: 185; Herv. i. Orig.): »Jemand, der nicht davon ausgeht, daß alles Daseiende ein Zug
in einem Sprachspiel […] sei, stößt schon weit unterhalb des god's point of view […] auf ein schwieriges Problem.«.
Oder um es mit Lave (1991: 67) auszudrücken: Die Kategorie ›Natur‹ ist genauso sozial generiert wie ›Nachmittagstee‹.
Vgl. insbesondere Abschnitt 3.5.4.
56
Erster Teil: Aufschluss
sich auf zwei unterschiedliche Seinsweisen beziehen. Die implizierte Subjektverdoppelung zu
erkennen, wäre somit der erste Schritt auf eine Beobachtertheorie hin. In der Konsequenz
heißt dies, dass ein Konstruktivismus arnoldscher Prägung, der Anleihen beim neurobiologischen Konstruktivismus nimmt, die Verwicklung in den Subjekt-Objekt-Dualismus beheben
kann, sofern er die Ehe mit dem Naturalismus und dem damit untrennbar verbundenen Bild
des ›homo clausus‹ aufgibt.44 Der kritische Bezug auf die Konstruktivismus-Varianten ist beispielhaft gemeint. Das heißt: Meine Kritik dient dazu, bis zu dem Punkt vorzustoßen, dass das
genannte Halt-Suchen an naturalistischen Vokabularen am leichtesten an diesen Theorien
aufzeigbar ist, aber auf ein grundlegenderes Problem hinweist, nämlich auf das Problem, dass
das Halt-Suchen am Bild des ›homo clausus‹ einen sozusagen immer wieder in die Bahnen
einer Ding- oder Substanzenontologie zwingt, welche einen sozialtheoretischen Dualismus zur
Folge hat. Diesen ›Mechanismus‹ näher darzustellen, gelten die weiter unten folgenden Abschnitte, denen man auch entnehmen kann, dass – in Ermangelung einer Alternative zum
›homo clausus‹ – auch ›gut gemeinte‹ Versuche, eine ›relationale‹ ›social theory of learning‹ zu
formulieren (vgl. Wenger 1998: 4, Lave/Wenger 1991) einer ontologischen Fundierung in
Form einer positiven Bestimmung der alternativen Menschen- und Sozialitätskonzeption bedürfen, sollen sie nicht auch in den Bannkreis einer Ding-Ontologie und des sozialtheoretischen Dualismus geraten.
Die Kritik am Repräsentationsmodell des Wissens im Rahmen konstruktivistischer Theorien richtet sich gegen »die erkenntnistheoretisch überkommene Position, dass Wirklichkeit im
Bewusstsein abbildbar und repräsentierbar sei« (Arnold 2007: 10). Sie wird auch als Zurückweisung einer ›Abbildtheorie des Geistes‹ (vgl. z. B. Arnold 2007: 57f., 62f., 68), eines Modells,
»das Erkennen als subjektives Abbilden objektiver Wirklichkeiten definiert« (Arnold/Siebert
1995: 88), von Abbild- und intellektualistisch verengten Informationsverarbeitungstheorien
(Arnold 1997a: 135) oder des Strebens nach »wahren« Erklärungen (Arnold 2003: 52) formuliert. Als erkenntnistheoretische Prämisse gilt hierbei, dass »der Mensch keinen unmittelbaren
erkenntnismäßigen Zugang zur Wirklichkeit hat« (Arnold 2003: 51). Insofern stelle sich ›der‹
Konstruktivismus als eine Subjekttheorie bzw. Theorie des erkennenden Subjekts dar, deren
Gegenstände »Prozesse der Wirklichkeitskonstruktion durch Beobachtung, nicht die Wirklichkeit selbst« (Arnold 2003: 52) seien.
Nun lautet für Rolf Arnold das Motto der erkenntnistheoretischen Position, dass Wirklichkeit im Bewusstsein abbildbar und repräsentierbar sei: »Es gibt eine Wirklichkeit außerhalb,
die wir erkennen und abbilden können!« (Arnold 2007: 10; i. Orig. kursiv) Die Phrase ›es gibt‹
zeichnet jedoch eine ontologische Aussage aus, sodass Arnold im Ergebnis den Unterschied
zwischen einem ontologischen Realismus und einem erkenntnistheoretischen Realismus verwischt. Erst auf Basis dieser Ausgangskonstellation kann dann eine pauschale Kritik an
(neo-)realistischen Positionen (z. B. Arnold 2005, 2007) formuliert werden und der Verzicht
auf »metaphysische Letztbegründungen« (Arnold/Siebert 2005: 62) eingefordert werden. So
gesehen gilt es hinter die Frage zu gelangen, ob ›der‹ Konstruktivismus selbst metaphysischrealistische Annahmen in sich trägt, wie Konstruktivismus-Kritiker meinen, oder ob NichtKonstruktivisten zwangsläufig einem metaphysischen Realismus das Wort reden.45
44
45
Für eine von neurobiologischen Theoremen gereinigte Fassung eines Konstruktivismus, welche als postkonstruktivistisch zu verstehen ist, vgl. die Arbeiten von Schmidt (2003a, 2003c), auf die hier lediglich ausblickend verwiesen werden kann (vgl. Abschnitt 7.1.2).
Einen Einblick in die wesentlichen Streitpunkte bieten z. B. Mitterer (1999) und Rustemeyer (1999). Freilich ist
dort die von mir im Weiteren vorgenommene Differenzierung nicht vorweggenommen.
1 Substanzialismus und Relationismus
57
Die von Arnold versäumte Differenzierung lässt sich gut anhand Gerhard Roths Sortierung des ›klassischen‹ Verständnisses von ›Realismus‹ und ›Idealismus‹ sowie an seiner Differenzierung zwischen ›Realität‹ und ›Wirklichkeit‹46 verdeutlichen. Darüber hinaus versuche ich
anhand der Rezeption von Roths Ansatz drei Punkte herauszuarbeiten. Erstens: Sofern zwei
›Welten‹ gegenübergestellt werden – eine nicht-erfahrbare ›Außenwelt‹ oder ›Realität‹ und eine
erfahrbare ›Innenwelt‹ oder ›Lebenswelt‹ oder ›Wirklichkeit‹ –, kann unter ›Ontologie‹ nur
Metaphysik verstanden werden. Und diese Art von Ontologie ist dualistisch. Zweitens: Da in
der neuzeitlichen Wissenschaft metaphysisches Denken ›verpönt‹ ist, versucht man nachmetaphysisch zu denken. Jedoch wiederholt sich das Innen-Außen-Schema auch in der Frage nach
der Struktur der »Wirklichkeit« (Roth), z. B. in der Frage, welcher »Wirklichkeitsbereich«
(Roth) der ›Ort‹ des Mentalen sei (z. B. die ›Außenwelt‹ im Sinne der ›Umwelt‹ des menschlichen Organismus oder die Innenwelt des menschlichen Organismus). Der Gedanke, dass eine
Wiederholung eines Grundmusters stattfindet, kommt auch bei Reckwitz (2004c: 320) zum
Ausdruck, wenn er mit Bezug auf Handlungserklärungen das Grundproblem einer »ontologischen Innen-Außendifferenz zwischen der ›geistigen Welt‹ (Husserl) und den äußeren Verhaltensakten« anspricht. Drittens: Die Frage nach der ›Struktur der Wirklichkeit‹ kann aber so
verstanden werden, dass nach den für Begriffe von ›Wirklichkeit‹ konstitutiven Sprachspielen
selbst gefragt wird. Ich werde zeigen, dass Roth bis zu diesem Punkt nicht vordringt und dass
daher im Anschluss an postempiristische Denkfiguren der Ansatz bei den gegenstandskonstitutiven Vokabularen erfolgen sollte. Im weiteren Fortgang der Darstellung wird die bisherige
Argumentation u. a. dazu genutzt werden, Themen wie ›Mentalismus‹ oder die Frage der
Handlungserklärung auszugliedern, den sozialtheoretischen Dualismus im Sinne eines Beschreibungsproblems zu fassen und sich von da aus mit Norbert Elias auf die Kernthese zu
kommen, dass die für den ›homo clausus‹ und ein substanzialisierendes Denken konstitutive
Mauer, die Grenze zwischen dem ›Inneren‹ und dem ›Äußeren‹ (Elias 1997a: 52f.) eingerissen
(›dekonstruiert‹) werden muss, um von da aus alternative »Denkmittel« (Elias; z. B. Elias
2003: 89, Elias 2004: 74) auffinden und verwenden zu können.
Roth geht es zum einen um den erkenntnistheoretischen Wert von Wahrnehmungen: Liefern sie Informationen über die Dinge der Außenwelt oder sind es Konstrukte? (vgl. zum Folgenden insbesondere Roth 1996: 339–344) Die Frage der klassischen Erkenntnistheorie lautet
nach Roth folglich: Ist objektives, allgemeingültiges Wissen bzw. objektive Erkenntnis (Wahrheit) möglich und, falls ja, in welchem Maße? Hieraus ergibt sich die Unterscheidung zwischen
einem Skeptizismus, welcher gesichertes Wissen für grundsätzlich unmöglich hält, und einem
erkenntnistheoretischen Idealismus bzw. Realismus, welche beide an die Möglichkeit gesicherten, objektiven Wissens glauben. Während der erkenntnistheoretische Realismus annimmt,
Sachverhalte der bewusstseinsunabhängigen Welt (›Realität‹) seien teilweise so zu erkennen,
wie sie tatsächlich sind, also von einer Erkennbarkeit der Realität (im Sinne einer Quelle gesicherten Wissens) ausgeht, nimmt der erkenntnistheoretische Idealismus an, wahres, objektives
Wissen könne nur aus dem Geist bzw. der Vernunft selber kommen.
Zum anderen steht die Frage im Raum, welchen ontologischen Status die Realität hat,
d. h., ob es sie gibt, und, falls ja, was man über sie aussagen kann. Während der ontologische
Idealismus (mitunter auch als ›objektiver Idealismus‹ bezeichnet), die Existenz einer bewusstseinsunabhängigen Welt leugnet, geht der ontologische Realismus von der Existenz einer bewusstseinsunabhängigen Welt aus, d. h., er nimmt Realität als unabhängig von der Existenz
des Menschen bzw. der Wahrnehmungstätigkeit existierend an.
46
Die ›Wirklichkeit‹, von der Arnold spricht, ist in Roths Terminologie ›Realität‹.
58
Erster Teil: Aufschluss
Durch eine Kreuztabellierung ergeben sich vier Kombinationen. Eine Kombination aus
ontologischem und erkenntnistheoretischem Realismus (1) arbeitet mit der Annahme, dass es
eine bewusstseinsunabhängige Realität gibt, welche zumindest teilweise erfahrbar ist. Da ein
erkenntnistheoretischer Realismus nach Roth vor der Frage steht, welches Wissen, das wir
über Erfahrung bzw. Wahrnehmung erlangen, aus der bewusstseinsunabhängigen Realität
stammt und welches durch den Menschen bzw. seinen Erkenntnisapparat beigegeben wird,
und da Roth die Abbildungsannahme47 bezweifelt, also die Annahme einer unmittelbaren
Widerspiegelung der Realität, verwirft Roth sowohl die Kombination aus ontologischem und
erkenntnistheoretischem Realismus als auch die Kombination aus erkenntnistheoretischem
Realismus und ontologischem Idealismus (2). Letztere ist, so hält Roth korrekterweise fest,
widersinnig, denn die Annahme einer Erkennbarkeit der Realität und die Leugnung der Existenz der Realität schließen sich aus (Roth 1996: 341). Roth verwirft insgesamt jeden erkenntnistheoretischen Realismus. Sowohl die Variante eines naiven Realismus, welcher annimmt,
dass der Wahrnehmungsapparat die Dinge exakt so abbilde, wie sie sind, als auch die abgeschwächte Variante eines kritischen Realismus, demzufolge einiges an den Wahrnehmungen
objektiv gegeben sei (objektive Erkenntnis), anderes subjektive Beigaben oder Hinzufügungen,
sind für ihn keine ernst zu nehmenden Positionen (vgl. Roth 1996: 342, 350, 353, 358). Die
Kombination aus ontologischem Idealismus und erkenntnistheoretischem Idealismus (3) arbeitet mit der Annahme, dass keine bewusstseinsunabhängige Welt existiert und wahres Wissen
via Vernunft möglich ist. Vertreter dieser Position sind z. B. Leibniz und Berkeley. Zur Vermeidung eines Solipsismus – nur ich existiere und alles andere ist nur Einbildung – nehmen
Berkeley und Leibniz an, dass die Sinneseindrücke von Gott eingegeben werden (vgl. Roth
1996: 341). Die Kombination aus ontologischem Realismus und erkenntnistheoretischem
Idealismus (4), die auch Roth vertritt, basiert auf der Annahme, dass eine bewusstseinsunabhängige Realität existiert, sie aber prinzipiell nicht erkennbar bzw. erfahrbar ist. Prototypisch
vertreten wird diese Position von Kant (in der ›Kritik der reinen Vernunft‹) und läuft auch
unter der Bezeichnung ›subjektiver Idealismus‹. Bei Kant existieren die ›Dinge an sich‹, sind
aber nicht erkennbar. Nur ihre Erscheinungen sind erkennbar. Die Dinge an sich lösen zwar
Sinnesempfindungen aus, aber die geordneten Wahrnehmungen und die Erkenntnis werden
als aller Erfahrung vorgeordnet und durch die ›apriorischen Anschauungsformen‹ und die
›Kategorien‹ geordnet angesehen (vgl. Roth 1996, v. a. 340f., 358).
Wie bereits erwähnt wurde, unterscheidet Roth zwischen »Realität« und »Wirklichkeit«
(vgl. Roth 1996: 314–338, Roth: 1987, Roth 1992b: 317–325). Die ›Realität‹ sei als eine objektive Welt bewusstseinsunabhängig, bewusstseinsjenseitig, unerfahrbar, unzugänglich und komme in der phänomenalen Welt nicht vor. Daher heißt sie bei Roth auch »transphänomenale
Welt« (z. B. Roth 1996: 316, 324f.). Die ›Wirklichkeit‹ hingegen ist für Roth die »Welt unserer
Empfindungen« (Roth 1996: 316) oder auch die »phänomenale Welt« (Roth 1996: 324).48 Sie
bestehe aus drei Bereichen, die Roth als »Wirklichkeitsbereiche« bezeichnet: die »Außenwelt«,
die »Welt des Körpers« und die »Welt unser geistigen und emotionalen Zustände« (Roth
1996: 314) bzw. die der »mentalen Zustände und des Ich« (Roth 1996: 314, 316). Diese drei
Bereiche seien normalerweise voneinander getrennt; beispielsweise seien Vorgänge in der
Außenwelt von Körpervorgängen wie auch von geistigen und emotionalen Zuständen ge-
47
48
Nach Skirbekk/Gilje (1993a: 412) kann der erkenntnistheoretische Realismus aufgrund der Abbildungsannahme
auch als repräsentativer Realismus bezeichnet werden, da er davon ausgeht, dass ›äußere‹ Dinge existieren und
dem Subjekt durch Sinneseindrücke, die die Dinge repräsentieren, zugänglich werden.
Den Terminus ›phänomenale Welt‹ leiht sich Roth von Wolfgang Köhler bzw. Wolfgang Metzger.
1 Substanzialismus und Relationismus
59
trennt, ebenso Körpervorgänge von geistigen und emotionalen Zuständen (vgl. Roth
1996: 314).
Roth weicht nun im Rahmen der Erörterung von Implikationen dieser Zweiweltentheorie
für seinen neurobiologischen Konstruktivismus keineswegs Fragen nach dem Verhältnis von
›Realität‹ und ›Wirklichkeit‹ aus, sondern hält einige wichtige Setzungen parat. Er unterscheidet
zwischen einem »realen Gehirn«, das wie der reale Körper, in dem es steckt, und die reale
Welt, in der dieser steckt, transphänomenal sei, und einem »wirklichen Gehirn« (Roth
1996: 329). Das reale Gehirn bringe die Wirklichkeit hervor (Roth 1996: 324). Das heißt: Die
neurologisch-physiologischen Prozesse des Gehirns lägen den mentalen Zuständen zugrunde
(z. B. Roth 1996: 317). Das Gehirn treffe in selbstreferenzieller Weise (aufgrund von Gehirnzuständen) die »Unterscheidungen über den Wirklichkeitscharakter erlebter Zustände« aufgrund von »Wirklichkeitskriterien« (vgl. Roth 1996: 321–324). Da die Realität unerfahrbar sei,
gelte, dass alle erlebten Vorgänge innerhalb der Wirklichkeit geschehen würden (Roth 1996,
z. B. 316, 325). Die dreigegliederte Wirklichkeit sei ein »Konstrukt des Gehirns« (Roth
1996: 317; Herv. i. Orig.). Hierbei betont Roth, dass das reale Gehirn das wirkliche Gehirn
hervorbringe, inklusive des erlebten ›Ich‹ – welches folglich ebenfalls ein Konstrukt des realen
Gehirns und nicht des wirklichen Gehirns sei (Roth 1996: 330) – sowie der Vorstellung, das
»Subjekt seiner mentalen Akte, Wahrnehmungen und Handlungen« (Roth 1996: 329; Herv. i.
Orig.) besitze einen Körper und stehe einer Außenwelt gegenüber (vgl. Roth 1996: 317,
329f.).49 In dem Konstrukt kommen, so Roth, die neurologisch-physiologischen Prozesse des
Gehirns nicht vor, wobei Roth meint, dass sie erlebnismäßig nicht erfahrbar sind (Roth
1996: 317). Schließlich sind sie ›reale‹ Vorgänge. Gleichwohl seien sie als Teile der Wirklichkeit
studierbar (vgl. Roth 1996: 317), mithin ein Teil der phänomenalen Welt und nicht Teil der
Realität (Roth 1996: 326).
Roth selbst stellt fest, dass man auf der Basis dieser Prämissen vor einer verwickelten Situation steht (Roth 1996: 332f.): Das Gehirn bringe Geist im Sinne von mentalen Zuständen
(d. h.: Geist ist Teil der Wirklichkeit) hervor; es könne aber nicht das wirkliche Gehirn sein.
Das wirkliche Gehirn, das dem Forscher zugänglich ist, bringe gar keinen Geist hervor. Dies
könne nur das reale Gehirn. Aber das sei unzugänglich. Folglich vollziehe sich Hirnforschung
nur innerhalb der Wirklichkeit und nicht innerhalb der Realität. Sie könne also keine Wesensschau betreiben, sondern nur die Phänomene der Wirklichkeit untersuchen und dort unter
bestimmte Qualitätsanforderungen stellen (z. B. Konsistenz; Plausibilität).
Jedoch hält Roth diesen aufscheinenden Widerspruch – auf der einen Seite die Behauptung, die Realität sei unerkennbar; auf der anderen Seite die Behauptung, man könne sich in
der Wirklichkeit auf sie beziehen – für scheinbar (Roth 1996: 359). Der Dreh- und Angelpunkt
für seine Argumentation in diesem Punkt ist die Aussage, dass die Realität eine Annahme sei
(Roth 1996: 324). Die Annahme einer bewusstseinsunabhängigen Welt, über die man gleichzeitig nichts erfahren könne (Roth 1996: 358), sei logisch notwendig: Um behaupten zu können, dass das reale Gehirn die Wirklichkeit erzeuge (vgl. Roth 1996: Kap. 13, z. B. Roth
1996: 325), müsse eine Entität bzw. eine Realität mit darin enthaltenen realen Gehirnen angenommen werden, »welche nicht selbst Teil der Wirklichkeit ist« (Roth 1996: 358).
49
Die Frage nach dem Status des Ich führt Roth zu der hinlänglich bekannten, äußerst kontrovers geführten und im
Weiteren hier nur sehr entfernt mitlaufenden Folgefrage, wem die ›Schuld‹ oder Verantwortung für Handlungen
zugeschrieben wird: dem Ich als Konstrukt oder dem realen Gehirn bzw. Organismus. Roth lässt hier keinen
Zweifel daran, dass er für einen Abschied von der Vorstellung vom Ich als Autor der Handlungen ist (vgl. Roth
1996: 329ff.).
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276
Les ḥiğāziyyāt sont arrivés jusqu’au Šām grâce au poète Muḥammad Ibn Naṣr, appelé al-Qayṣarānī (m. 548h/1153). Mais bien avant cela, ses poésies ont atteint alAndalus, après avoir dédié plusieurs poèmes à Ismāˁīl Ibn ˁAbbād, émir des Abbadides, car ce dernier était très intéressé par al-Raḍī. Le voyage de la poésie de Šarīf a donc commencé dès l’époque même de Šarīf. Vingt ans après sa mort, son divan était partout en al-Andalus, influençant Ibn Šuhayd (m. 426h/1035), intégrant deux vers de la poésie d’al-Raḍī dans la sienne 699. Cela montre que les ḥiğāziyyāt se sont répandues dès le début auprès des poètes de l’époque des Ṭawāˀif. Ils ont intégré le genre des ḥiğāziyyāt dans la première partie de leur poésie, à la manière du nasīb, avant d’en faire un genre complet pour des poésies indépendantes. Ainsi, le poète Ibn al-Muˁallim, un des vizirs de Muˁtaḍid Ibn ˁAbbās, montre sa passion pour le Nağd en utilisant les moyens connus chez Šarīf, comme l’éclair du Nağd. Ou encore, Ibn al-Ḥaddād al-Andalusī (m. 480h/1087), dans un panégyrique pour Muˁtasim Ibn Ṣamādiḥ, cite les lieux du Ḥiğāz : « Et Taymāˀ, pour le cœur amoureux, est une station Alors retournez avec des salutations pour ses Salumāt. » Puis il poursuit, un peu plus loin : « Des sentiments d’amour ardent et la Kaˁba des épreuves Mon cœur est parmi ses pèlerins et ses suppl ants Combien de fois m’a serré la main de l’espoir à Mina Et combien de fois a soufflé l’odeur de la joie de ses ˁArafāt 700? » Nous remarquons que ces images, les lieux cités, cette distinction entre Minā et munā, l’espoir, les Salumāt, la passion pour ces lieux saints (même sans avoir vu ces lieux), relèvent d’une méthode reprise de Šarīf al-Raḍī, d’ailleurs régulièrement reprise par les 699 Aḥmad Ḥāğim al-Rubayˁī, Al-Ġurba wa-l-Ḥanīn fī al-Šiˁr al-Andalusī, op. cit., p. 86. 700 Ibn al-Ḥaddād al-Andalusī, Dīwān Ibn al-Ḥaddād al-Andalusī, Beyrouth, Dār al-Kutub al-ˁIlmiyya, 1990, no 6, p. 162 (ṭawīl) : « wa-taymāˀu li-l-qalbi l-mutayyami manzil
un
fa-
ˁ
ū
ğā bi-tas
lī
min ˁalā salamāti-hā mašāˁiru tihyāmin wa-kaˁbatu fitnatin fuˀādiya min ḥuğğāği-hā wa-duˁāti-hā fa-kam ṣāfaḥat-nī fī-minā-hā yadu l-munā wa-kam habba ˁarfu l-lahwi min ˁarafāti-hā. » 277 poètes qui l’ont suivi
, peut-être parce que le Ḥiğāz est considéré comme la terre d’origine et que cette région est le centre des lieux saints, ce qui les a amenés à se sentir étrangers. Un autre poète, Ibn al-Labbāna al-Dānī (m. 507h/1113), reprend l’image de l’éclair pour éveiller la nostalgie, quand il dit qu’avec le vent de ṣabā et l’éclair lumineux du Ḥiğāz, le ḫayāl et l’imagination du Ḥiğāz : « Il désire al-ˁAqīq et ses habitants, même si Les nouvelles de ˁAqīq et ses habitants sont passés 701. » Après l’époque des Ṭawāˀif, à l’époque des Murābiṭīn, les ḥiğāziyyāt étaient très présents, dans tous les prologues des poésies. Rubayˁī explique : « Cela est dû à la diffusion du divan de Šarīf al-Raḍī entre eux et du divan des poètes qui l’ont suivi sur sa voie, comme Mihyār al-Daylamī, Ṣardar al-Ṣūrī, parmi d’autres 702. » Les lieux du Ḥiğāz ont même tellement été cités qu’un des poètes, ˁAbdullāh Ibn Wāğib, s’est mis à dire que cela sufisait et qu’il était temps d’arrêter de citer ces lieux : « Cessez de mentionner Naˁmān, Salˁ et Ġarb 703! » D’après lui, les poètes avaient exagéré, alors qu’un grand nombre d’entre eux n’avaient même pas vu ces lieux et expérimenté les rituels qui leur sont liés, non pas comme Šarīf al-Raḍī, qui avait une expérience directe du hadj, qui n’était pas un événement purement imaginaire pour lui, et sa passion véritable découlait de son vécu, ce qui le rend unique. Ibn Ḥamdīs al-Ṣiqillī (m. 527h/1133) et Ibn Zaqqāq al-Balansī (m. 530h/1136), neveu d’Ibn Ḫafāğa, mentionnent également beaucoup les lieux du Ḥiğāz. Ibn Ḫafāğa luimême, dans l’introduction de son divan, déclare clairement qu’il a été influencé par Šarīf al-Raḍī, al-Ṣūrī et Mihyār al-Daylamī 704. D’ailleurs, 701 Abū Bakr Ibn al-Labbāna al-Dānī, Dīwān Ibn al-Labbāna al-Dānī, Amman, Dār al-Rāya, 2008, p. 81 v. 5 (kāmil) : « yahwa-l-ˁaqīqa wa-sākinī-hi wa-in yakun ḫabaru l-ˁaqīqi wa-sākinī-hi qad inqaḍā. » 702 Aḥmad Ḥāğim al-Rubayˁī, Al-Ġurba wa-l-Ḥanīn fī al-Šiˁr al-Andalusī, op. cit., p. 88 : « Wa-ḏālika li-šuyūˁi dīwāni š-šarīfi r-raḍiyyi bayna-hum wa-dawāwīni š-šuˁarāˀi l-laḏīna sārū ˁalā-nahğihi miṯli mahyār al-daylamī wa-ṣardar wa-ṣ-ṣūrī wa-ġayri-him. » 703 Ibid
, p.
89
:
« an –yatrukū ḏikra naˁmāna wa-salˁin wa-ġarbin. » 704
Brigitte Foulon, La poésie andalouse du XIe siècle
,
op
.
cit
., p.
18.
278 « Ibn Ḫafāğa n’était pas le seul à être influencé par le style “arabe” ou bédouin, qui jouissait d’une très grande popularité en al-Andalus. Cette poésie “arabe était vécue par les Andalous par le biais de poètes orientaux contemporains qui imitaient et préconisaient le style bédouin. Al-Mutanabbī et les ḥiğāziyyāt d’al-Šarīf al-Raḍī étaient particulièrement populaires 705. » Ce poète qu’est Ibn Ḫafāğa est attaché à ces lieux, répétant même les noms du Ḥiğāz dans des poèmes autres que ses ḥiğāziyyāt, sans y être allé pour autant. Il cite notamment des lieux de chemins de la Mecque, depuis l’Irak, comme Laˁlaˁ, un lieu qui se trouve entre Kūfa et Bassorah : « Ô odeur d’un vent qui revient de la vallée de Laˁlaˁ Qui traîne sur la terre avec de la rosée les rêves usés des chameaux Avec la distance qui nous sépare, à Ḥaqf, du désert de ˁĀliğ Et au croisement d’Arṭā à l’avant-mont de Šimām 706. » Dans cette ḥiğāziyya de type thrène, Ibn Ḫafāğa commence avec le vent de salutations et l’éclair qui cause le feu de la passion. Puis il cite des noms de lieux et il évoque aussi la jeunesse et la vieillesse, deux images connues chez Šarīf. Ce qu’il est également intéressant de voir, c’est qu’il a lié Ġamīm à l’Euphrate 707, ce qui peut confirmer que Ġamīm se situe en Irak. Reprenant encore des images de Šarīf, Ibn Ḫafāğa se demande ce qui appelle la passion. Serait-ce l’éclair ou bien le chant de la colombe? Puis il évoque la vallée de Ḏī al-Ġaḍā et transmet ses salutations à ces lieux. Dans ce poème comme dans d’autres, bien des toponymes du Ḥiğāz sont cités. En expliquant la raison, il dit dans son divan que les noms de ces lieux, d’ entre les lieux du Nağd et d’autres terres, sont des 705 Magda M. al-Nowaihi, The Poetry of Ibn Khafajah: A Literary Analysis, New York, E.J. Brill, 1993, p. 3-4. 706 Ibrāhīm Ibn Abī al-Fatḥ Ibn ˁAbdullāh Ibn Ḫafāğa al-Andalusī, Dīwān Ibn Ḫafāğa, annoté par ˁAbdullāh Sanda, Beyrouth, Dār al-Maˁrifa, 2006, no 26, p. 308 (ṭawīl) : « fa-yā ˁarfa rīḥin ˁāğa-ˁan baṭni laˁlaˁin yağurru ˁalā l-andāˀi faḍla zimāmī bi-mā bayna-nā bi-l-ḥaqfi min ramli ˁāliğin 707 Ibid., p. 307, v. 7 (ṭawīl) : « wa-rubba layālin bi-l-ġamīmi ariqtu-hā wa-fī multaqā ll-arṭā bi-safḥi šimāmī. » li-marḍā ğufūnin bi-l-furāti niyāmī. » 279 symboles et des exemples qui renvoient à ce qui leur ressemble sans les citer 708.
Les noms cités par Šarīf al-Raḍī sont donc devenus des symboles auxquels on s’identifie, chacun dans sa culture. Les toponymes renvoient aux aspects de ces lieux, aux émotions qui leur sont liées et à leurs charges. Aḥmad al-Rubayˁī remarque : « L’appel d’Ibn Ḫafāğa au chant des lieux du Ḥiğāz, c’était une façon de rappeler les villes andalouses, qui sont devenues chez lui aussi des symboles sacrés, comme les lieux du Ḥiğāz, qui n’étaient pas vides en habitants ou divertissements, mais c’étaient des symboles qui avaient une vie propre. Et ainsi, il les emploie pour attirer le cœur vers lui ainsi que les oreilles 709. » Ce qui a attiré Ibn Ḫafāğa chez Šarī , c’est la vie, les vestiges n’étant plus des lieux morts, sans vie justement, Šarīf insistant le premier sur les lieux du Ḥiğāz en tant que lieux où il a vécu des expériences personnelles. Pour al-Raḍī, ces lieux sont de vrais lieux qu’il connaît au-delà de l’imaginaire, mais il ouvre une porte de symboliques pour utiliser ces lieux, même sans les connaître, en tant que symboles. L’exemple mentionné de Ġamīm semble assez emblématique de la situation, ce lieu étant difficile à déterminer, mais il s’est totalement transformé en symbole que les poètes pouvaient à nouveau utiliser, sans forcément connaître sa géographie avec précision. Ṣafwān Ibn Idrīs al-Mursī (m. 598h/1202) considère les citations des lieux du Ḥiğāz comme une question d’imagination, et il avoue ne pas avoir eu l’expérience de la vie dans le Nağd et qu’il n’a pas de passion spécifique pour le Nağd, mais qu’il ne faisait que reprendre une méthode qui a besoin du Nağd pour embellir la poésie : l’évocation du Nağd et du Ḥiğāz, dans le livre de la poésie, est comparable à la sourate « Al-Ḥamd » pour le Coran 710, c’est-à-dire que c’est devenu l’introduction inévitable. À l’époque de Banī al-Aḥmar, les Nasrides de Grenade, en al-Andalus, dans les années 700h/1300, quand les villes subissaient défaite après défaite dans cette région, la mention du Ḥiğāz avait pris une tournure politique. Le Ḥiğāz était devenu un synonyme 708 Ibid., p. 308, v. 15. 709 Aḥmad Ḥāğim al-Rubayˁī, Al-Ġurba wa-l-Ḥanīn fī al-Šiˁr al-Andalusī, op. cit., p. 91-92 : « kānat daˁwatu bni-ḫafāğata li-t-taġannī bi-d-diyāri l-ḥiğāziyyati namaṭan mina-t-taḏkīri bi-l-muduni landalūsiyyati l-latī tahawwalat laday-hi ilā-rumūzin muqaddasatin miṯlu-hā miṯlu d-diyāri l-ḥiğāziyyati llatī lam-takun ḫāliyatan min-ahli-hā ˁāṭilatan ˁan-awānisi-hā bal-hiya rumūzun tanbuḍu bi-l-ḥatāti wa-biḏālika yasūġu īrāda-hā li-yastamīla l-qulūba ilay-hi wa-yağḏiba l-iṣġāˀa naḥwa-hu. » 710 Ibid., p. 93. 280 de résistance. Chaque ville d’al-Andalus qu’ils perdaient était vue comme une ville sainte qui tombait. Ainsi, les toponymes du Ḥiğāz étaient des symboles directs pour parler de ces lieux, de ces villes. Par exemple, sous la plume d’Ibn al-Hakīm al-Laḫmī (m. 708h/1308), al-Liwā était devenue un symbole pour parler de Grenade. Abū Ḥayyān (m. 745h/1344), quant à lui, cherchait à évoquer la nostalgie pour les lieux, se servant alors de l’éclair et de la brise. Ibn al-Ḫaṭīb (m. 776h/1374) a lui, de son côté, été grandement influencé par Šarīf al-Raḍī. ˁIṣam Qasabji, en parlant de lui, remarque que dans ses poèmes au long nasīb, on trouve des signes symboliques qui renvoient aux lieux du Ḥiğāz, des lieux de beauté et de spiritualité, et que se répètent les champs lexicaux de l’ascétisme et la patience, comme pour parler du bien-aimé qui tourne le dos. Cela confirme le grand lien entre ses sentiments du cœur et ses sentiments religieux, d’une façon qui nous rappelle Šarīf al-Raḍī 711. Dans ses images, Ibn al-Ḫaṭīb parle de la chamelle (al-nāqa), à qui il veut faire sentir l’odeur du Nağd et du Ḥiğāz, une terre que même sa chamelle désire : « Vous deux, laissez-la sentir les traces du Nağd, car dans le Nağd se trouve Une passion, dont est sorti avec le souvenir un amour caché 712. » Cette chamelle ressemble fortement à celle d’al-Raḍī, qu’il a habituée au sable du Ḥiğāz 713. Et al Rubayˁī en vient même à dire que l’influence de Šarīf al-Raḍī est très claire sur le roi de Grenade, Yūsuf III (m. 819h/1416), qui reprend des extraits de sa poésie dans la sienne, à tel point qu’il a repris la gazelle du saule 714, une marque de fabrique de Šarīf. Comme nous l’avons vu au préalable, les ḥiğāziyyāt ont également exercé une grande influence auprès des poètes soufis. En orient, Abū Saˁīd Abū al-Ḫayr (m. 441h/1049), considéré parmi les premiers poètes soufis au Ḫurāsān, reprend des 711 Hayṯam al-Ğarūd, Taṭawwur Fann al-Ḥiğāziyyāt Ilā Nihāyat al-Qarn al-Hiğrī al-Sābiˁ, mémoire sous la direction de Muḥammad Ḥamawiyya et ˁĪsā al-ˁĀkūb, op. cit., p. 5. 712 Lisān al-Dīn Ibn al-Ḫaṭīb al-Salmānī, Dīwān Lisān al-Dīn Ibn al-Ḫaṭīb, annoté par Muḥammad Miftāḥ, op. cit., T. 1, p. 300, v. 1, (ṭawīl) : « daˁā-hā tašim āṯāra nağdin fa-fī-nağdin 713 Dīwān, T. 2, p. 31, v. 6 (ṭawīl) : « wa-ašmamtu-hā ramla l-unayˁimi ġudwatan hawan hāğa min-hā ḏikru-hū kāminu l-wağdī. » fa-sāfat bi-anfin munkirin ġayri ˁārifī. » 714 Aḥmad Ḥāğim al-Rubayˁī, Al-Ġurba wa-l-Ḥanīn fī al-Šiˁr al-Andalusī, op. cit., p. 102. 281 images des ḥiğāziyyāt, notamment celle de la femme et de la nuit, dans ses poésies mystiques : « Hier soir, j’ai parlé avec le bien-aimé Des mots d’amour et de communion Il a commencé à rompre les pactes Et il s’en prend à moi par caprice Et la nuit est passée sans que je ne parle De l’histoire de l’amour si ce n’est son début Et la nuit n’est pas fautive, mais cependant L’histoire de l’amour n’a pas de fin 715. » Ce quatrain n’est pas loin du poème commençant par « Ô toi, nuit de l’avant-mont ». Chez ces deux poètes, nous retrouvons l’amour spirituel exprimé à travers les motifs de l’amour des femmes. Chez les soufis toujours, Abū Madyan al-Tilimsānī (m. 594h/1198), un des chefs soufis au Maghreb, a joué un rôle dans l’intégration de certains aspects des ḥiğāziyyāt dans les poésies mystiques : « Des gens disent que la passion s’est emparée de lui Oui! Je ne suis pas le premier à devenir fou de Laylā. » Ce poète poursuit plus loin : « Le chamelier a appelé son nom quand nous étions à Ġaḍā Nous lui avons alors dit : Mentionne-la encore davantage pour nous. » Encore plus loin, il continue : 715 ˁĀṭif Ğawdat Naṣr, Al-Ramz al-Šiˁrī ˁInda al-Ṣūfiyya, op. cit., p. 167 : « laylatu l-amsi ḥaddaṯtu l-ḥabība aḥādīṯa l-ḥubbi wa-l-wiṣālī Fa-aḫaḏa yanquḍu l-ˁuhūda wa-yatağannā ˁalayya fī-dalālī wa-nqaḍati l-laylatu wa-lam aḥki min qiṣṣati l-ḥubbi illa l-bidāya wa-lā ḏanba li-l-layli wa-lākin qiṣṣatu l-ḥubbi laysa la-hā nihāya. » 282 « Même les branches du saule se penchent Et sur chaque branche, un oiseau a chanté une mélodie 716. » Dans ces vers, nous ressentons l’influence des ḥiğāziyyāt sur ce célèbre poète maghrébin, passionné par le chemin du pèlerinage, même si ce n’est pas un pèlerinage direct, physique. En effet, pour certains soufis, le hadj important est spirituel et intérieur, la volonté étant simplement de se rapprocher de Dieu. Mais dans les images, comme nous le voyons dans les vers susmentionnés, il profite de l’amour de Laylā avec la brise et le chamelier, les liant à un lieu du Ḥiğāz, al-Ġaḍā, et la passion est transférée aux chameaux et aux branches des saules, ainsi qu’aux oiseaux qui se tiennent perchés sur ces branches. Tout cela nous rappelle les poésies d’al-Raḍī, ce qui n’est pas surprenant, vu l’aspect mystique largement présent dans ses ḥiğāziyyāt. À l’Est, un autre poète irakien, Ibn al-Muˁallim al-Wāṣiṭī (m. 592h/1196), considéré parmi les poètes à avoir influencé directement Ibn al-Fāriḍ, était un poète aux poésies douces et délicates d’après Ibn Ḫallikān, affirmant qu’il est de ceux dont la poésie a voyagé, est devenue réputée et dont le niveau est très élevé, et dont la poésie d’amour est très proche du cœur 717. Sa réputation est certainement due à ses disciples, qui chantaient et répétaient tout de suite ses poèmes dès qu’ils faisaient leur apparition. La répétition augmente logiquement l’attachement à ces poésies, et donc leur réputation. Ce poète a énormément été influencé par Šarīf al-Raḍī, surtout par l’aspect doux de l’amour lié à la douleur, la souffrance, avec finalement le paradoxe entre la joie et les douleurs : « Je jure, par ce sur quoi leurs lèvres se sont fermées Sur l’eau fraîche dans des perles cachées 716 Ibid., p. 168 : « taqawwala nāsun qad tamallaka-hu l-hawā [...] ağal lastu fī-laylā bi-awwali man ğunnā daˁā bi-smiha l-ḥādī wa-naḥnu ˁala l-ġaḍā [...] fa-qulnā la-hū bi-l-lāhi min ḏikri-hā zidnā wa-ḥattā ġuṣūnu l-bāni mālat tarannuḥan wa-ġannat ˁalay-hā kullu ṣādiḥatin laḥnā. » 717 Muṣṭafā al-Šībī, « Hiğāziyyāt al-Šarīf al-Raḍī », Al-Šarīf al-Raḍī Dirāsāt fī Ḏikrāh al-Alf
, op. cit., p. 49. 283
Si le chamelier arrive à ˁ
Uḏayb
,
je
mourrai Et qui peut m’aider pour que mon serment se produise? Sans les traces de Laylā et de la passion
Dans
ses collines, je ne serais pas parti comme Mağnūn
718
. » L’image du chamelier qui arrive en un lieu précis, ˁUḏayb, et qui montre sa passion liée à ce lieu du Ḥiğāz, rappelle directement les écrits d’al-Raḍī. Pour aller plus loin, d’après al-Šībī, le poème d’Ibn al-Muˁallim dont proviennent les vers suivants est le meilleur lien entre Šarīf al-Raḍī et Ibn al-Fāriḍ, faisant fortement entrer les ḥiğāziyyāt dans le soufisme : « Renvoyez-moi les groupes perdus Si la maison est dépeuplée, ce n’est pas ma patrie. » Un peu plus loin, il est dit : « Ô habitants de Naˁmān, où est notre époque À Ṭuwayliˁ, ô habitants de Naˁmān 719? » Ibn al-Fāriḍ (m. 632h/1235), grand poète soufi, a pour sa part fait entrer les ḥiğāziyyāt dans sa poésie et dans la poésie soufie en général. Nous retrouvons chez lui l’esprit de Šarīf al-Raḍī avec les mots, les images et les toponymes. Ibn al-Fāriḍ n’était pas un poète naturel comme Šarīf, puisqu’il essayait de tisser sa poésie, sans que cela ne fasse perdre d’intérêt pour ses écrits, dans lesquels le sens de la passion pour les lieux est bien présent. Il est à savoir que ce poète, à l’instar de Šarīf, a vécu au Ḥiğāz, puisqu’il y est resté près de quinze ans, tout proche de Médine en particulier . Il parle alors de son expérience dans les vallées de la Mecque et ses montagnes, où il prenait plaisir à voir les bêtes sauvages 718 Abū al-ˁAbbās Šams al-Dīn Aḥmad Ibn Muḥammad Ibn Abī Bakr Ibn Ḫallikān, Wafayāt al-Aˁyān, op. cit., T. 5, p. 7 : « qasaman bi-mā ḍummat ˁalay-hi šifāhu-hum min-qarqafin fī-luˀluˀin maknūnī in-šārafa l-ḥādi l-ˁuḏayba la-aqḍiyan naḥbī wa-man-lī an-tabarra yamī-nī law-lam yakun āṯāru laylā wa-l-hawā 719 Ibid., T. 5, p. 6 : « ruddū ˁalayya šawārida l-aẓˁānī [...] yā-sākinī naˁmāna ayna zamānu-nā bi-tilāˁi-hī mā-ruḥtu ka-l-mağnūnī. »
ma-d
-dāru in-lam taġna min-aw
ṭ
ānī bi
-
ṭuwayli
ˁ
in ya
-
sākinī
naˁmā
nī
.
» 284
jour et nuit. Il a en effet également résidé dans une vallée à dix jours de marche de la Mecque 720. Mais à la fin de sa vie, il est retourné en Égypte, où il a ressenti la nostalgie pour les lieux saints, et c’est à ce moment-là que nous ressentons le plus le souffle de Šarīf al-Raḍī : « Est-ce que le feu de Laylā est apparu la nuit de Ḏī Salam? Ou l’éclair est-il apparu à Zawrāˀ puis ˁAlam? Les brises de Naˁmān ne peuvent-elles souffler pour nous avant l’aube? Et l’eau de Wağra ne peut-elle être bue à petites gorgées? Ô chamelier du groupe qui parcourt les déserts, contraignant Comme on parcourt un livre à l’endroit des armoises à Iḍam. » Il poursuit plus loin : « Et stationne à Salˁ, et demande à Ğizˁ s’il a plu Sur les arbres de Aṯīl à Raqmatayn. » Puis il dit encore : « Je te demande, par Dieu, si tu passes à ˁAqīq la matinée De leur transmettre les salutations sans être gêné. » Encore plus loin, il continue : « Ô gazelles qui se retournent, pour votre statut J’ai su que mon œil ne regarde personne d’autre Je suis au service d’un juge qui a donné un étrange verdict Il a décidé de verser mon sang dans le ḥill et le ḥaram 721. » 720 Ibn al-Fāriḍ, Šarḥ Dīwān Ibn al-Fāriḍ, annoté par Ḥasan al-Būraynī et ˁAbd al-Ġanī al-Nābulsī, Ḥanān Abī Ṭāqiya (Egypt), Al-Maṭbaˁa al-ˁĀmira al-Šarīfa, 1888, p. 5. 721 Abū Ḥafṣ Šaraf al-Dīn ˁUmar Ibn ˁAlī Ibn Muršid al-Ḥamawī, Dīwān Ibn al-Fāriḍ, Beyrouth, Dār Ṣādir, 2011, p. 128 et 129, v. 1-3 et 5-6 et 16-17 (basīṭ) : « hal nāru laylā badat laylan bi-ḏī salamī am-bāriqun lāḥa fi-z-zawrāˀi fa-l-ˁalamī 285
Cette poésie, dont nous venons de citer un certain nombre de vers, laisse très clairement transparaître l’esprit des poésies d’al-Raḍī. Ibn al-Fāriḍ semble être comme le disciple direct de Šarīf, malgré les quelque deux cents années qui les séparent. Dans les mots utilisés par Ibn al-Fāriḍ, nous retrouvons le feu (nār), les larmes, les gazelles, les caravanes, l’ambivalence entre la joie et la douleur, l’eau, et surtout le lieu Ḏī Salam, bien connu chez Šarīf al-Raḍī, et encore d’autres mots du vocabulaire de Šarīf, comme ḥill et ḥaram, l’éclair (bāriq), Laylā, ˁAqīq, les jours de Ḫayf, Wağra, Naˁmān, Iḍam, etc. Dans une autre poésie, Ibn al-Fāriḍ cite le lieu Ḥāğir de Šarīf al-Raḍī, qui est un lieu dans le désert du Ḥiğāz sur la voie du hadj 722. Al-Raḍī a mentionné Ḥāğir et Maḥāğir plusieurs fois, et les deux sont repris par Ibn al-Fāriḍ dans le premier vers d’un poème : « Protège ton cœur si tu es passé par Ḥāğir Ses gazelles sont comme l’épée dans les yeux (Maḥāğir) 723. » Ce lieu, Ḥāğir, que Šarīf a aimé et rattaché à l’armoise, est intéressant chez Ibn al-Fāriḍ, du fait qu’il ait réussi à combiner Ḥāğir et Maḥāğir, avec l’image reprise de Šarīf : « La nuit nous a donné l’occasion de voir les beaux visages Et parmi les flèches, les yeux et leurs contours 724. » Et dans cette poésie d’Ibn al-Fāriḍ, nous retrouvons une partie du lexique de Šarīf, comme kaṯīb, ṭayf et zawr. Ce poète, plus proche du métier d’Abū Tammām, essaye de jouer avec les mots, ce que Šarīf faisait naturellement, sans exagération. Ibn al-Fāriḍ a cette tendance arwāḥa naˁmāna hallā nasmatun saḥaran yā-sāˀiqa ẓ-ẓaˁni yaṭwi l-bīda muˁtasifan wa-māˀa wağrata hallā nahlatun bi-famī ṭayya s-siğilli bi-ḏāti l-šīḥi min iḍamī [...] wa-qif bi-salˁin wa-sal bi-l-ğizˁi hal muṭirat bi-r-raqmatayni uṯaylātin bi-munsağimī nā dtuka l-lāha in ğuzta l-ˁaqīqa ḍuḥan [...] fa-qra s-salāma ˁalay-him ġayra muḥtašimī ˁannī ilaykum ẓibāˀa l-munḥanā karaman ˁahdtu ṭarfiya lam-yanẓur liġayri-himī ṭawˁan li-qāḍin atā fī-ḥukmi-hī ˁağaban aftā bi-safki damī fi-l-ḥilli wa-l-ḥaramī. » 722 Ibn Manẓūr, Lisān al-ˁarab, op. cit.,
T
. 4, p.
42. 723 Abū Ḥafṣ Šaraf al-Dīn ˁUmar Ibn ˁAlī Ibn Muršid al-Ḥamawī, Dīwān Ibn al-Fāriḍ, op. cit., p. 148, v. 1 (basīṭ) : « iḥfaẓ fuˀādaka in-mararta bi-ḥāğirī 724 Dīwān, T. 2, p. 414, v. 4 (kāmil) : « sanaḥat la-nā fī-l-mušriqāti ˁašiyyatan fa-ẓibāˀu-hū min-hā ẓ-ẓubā bi-maḥāğirī. » wa-mina s-sihāmi maḥāğirun wa-ˁuyūnū. » 286 à reprendre des mots ou des images, utilisant le procédé de
paronomase
(ğinās), comme avec l
’éclair
(
barq
ou
b
āri
q).
Dans un autre poème
,
il
dit : « Est-ce que l’éclair est apparu du côté de Ġawr, éclatant? Ou est-ce la burqa qui est soulevée du visage de Laylā? Ou est-ce le feu de Ġaḍā qui a éclairé alors que Salmā est à Ḏī al-Ġaḍā? Ou a-t-il souri devant ce que les yeux lui ont raconté 725? » Puis il continue à citer des lieux spécifiques de Šarīf, comme Laˁlaˁ, alors que Laˁlaˁ n’est pas sur son chemin, puisqu’Ibn al-Fāriḍ voyageait entre le Ḥiğāz et l’Égypte. Laˁlaˁ est répété deux fois dans une même partie de vers, et il cite aussi Ḥāğir, Waˁsāˀ, Naqā, Nağd, Kāẓima, les Salumāt dans le Ḥiğāz, Ğizˁ, ˁĀliğ, Raqmatayn, Ġuwayr, Ḍāriğ, ˁĀmir, Maˀzamayn, Qibāb, Ğamˁ, Ḫayf, al-Ḥiğr, Zamzam et la Mecque. Il est intéressant de voir qu’il a choisi de mentionner la caravane irakienne, comme un clin d’œil à Šarī f al-Raḍī. Dans toutes ses poésies, nous ressentons l’influence déterminante d’al-Raḍī, avec les lieux, les arbres, les expériences, les chameaux et surtout les images qui les accompagnent. « Ibn al-Fāriḍ dans ses poèmes, [...], a utilisé l’essence féminine et, a décrit, par les sentiments humains et la beauté terrestre éphémère, son amour divin et son désir tourné vers la beauté dans sa hauteur et l’absolu, en distinguant dans cette description poétique la beauté réelle liée à l’unicité, la totalité et l’éternité d’une part, de la beauté métaphorique d’autre part 726. » En fait, la différence entre Ibn al-Fāriḍ et Šarīf al-Raḍī, c’est l’étiquette soufie que porte le premier. Ayant pris ce qualificatif, sa poésie est également qualifiée de soufie. Changeant ainsi de catégorie, la poésie est vue d’un œil différent, sous de nouveaux 725 Abū Ḥafṣ Šaraf al-Dīn ˁUmar Ibn ˁAlī Ibn Muršid al-Ḥamawī, Dīwān Ibn al-Fāriḍ, op. cit., p. 166 v. 1 (ṭawīl) : « A barqun badā min-ğānibi l-ġawri lāmiˁū anāru
l-ġaḍā
ḍāˀ
at wa-salmā bi-ḏī l-ġaḍā am-irtafaˁat ˁan-wağhi laylā l-bar
ā
qiˁū
ami-btasamat ˁammā ḥakat-hu l-madāmiˁū
. »
726
ˁĀṭif Ğawdat Naṣr, Al-Ramz al-Šiˁrī ˁInda al-Ṣūfiyya, op. cit., p. 174 : « fī-ašˁāri-bni-l-fāriḍi, [...], ahāba fī-hā bi-l-ğawhari l-unṯawiyyi iḏ-waṣafa min-ḫilāli l-ˁawāṭifi linsāniyyati wa-l-ğamāli l-arḍiyyi z-zāˀili, ḥubba-hu l-ilāhiyyā wa-taˁaššuqa-hu bi-l-ğamāli fī-ˁuluwwi-hi wa-iṭlāqi-hi, mumayyizan fī-hāḏā l-waṣfi š-šiˁriyyi bayna l-ğamāli l-ḥaqīqiyyi l-muttasimi bi-l-waḥdati waš-šumūli wa-l-abadiyyati, wa-l-ğamāli l-mağāziyyi. » 287 projecteurs, notre regard porté sur telle ou telle poésie dépendant pour beaucoup du genre dans lequel elle a été classée. Ibn al-Fāriḍ, comme nous l’avions vu pour Ibn ˁArab , explique de lui-même que ses propos sont employés pour parler de l’amour de Dieu 727. Il se met lui-même dans une catégorie particulière en tant que poète, et il met donc sa poésie dans la catégorie soufie. Ainsi, le regard des gens est porté sur cette poésie comme étant le fruit d’une réflexion voire d’une inspiration mystique. La poésie, à l’origine œuvre littéraire de l’esprit humain, revêt tout à coup un caractère sacré. À la différence de Šarīf al-Raḍī et ses ḥiğāziyyāt, ce ne sont plus que les lieux mentionnés qui sont sacrés de par leurs sens spirituels, mais toute la poésie est vue comme sacrée. Cette sacralisation de la poésie retire d’une certaine façon toute ambiguïté sur les sens de l’amour et donne lieu à bien des analyses qui tentent de donner des sens mystiques à chaque mot choisi, comme barq, nasīm, ṣabā, ṣafā et le hadj, qui est devenu l’intention de partir vers Dieu. Malgré la présence claire de l’aspect spirituel chez Ibn al-Fāriḍ, certains se demandent pourquoi ces lieux ne sont pas simplement vus comme des lieux qu’il a aimés après les avoir vus et y avoir vécu, comme Šarīf al-Raḍī, surtout en étant resté longtemps dans le Ḥiğāz. Mais d’autres commentateurs ont insisté sur l’aspect symbolique, donnant des interprétations pour que ces lieux soient vus comme des signes et des métaphores 728. Finalement, l’expérience du hadj, chez Šarīf al-Raḍī comme par la suite chez Ibn al-Fāriḍ, se traduit à chaque fois en un sentiment spirituel exprimé par des moyens poétiques physiques ou sensuels. Ibn ˁArabī (m. 638h/1240) est un autre soufi célèbre influencé par Šarīf al-Raḍī, que nous avons déjà cité plus tôt. Il est connu pour son amour pour Niẓām, appelée aussi ˁAyn al-Šams (l’œil du soleil), la fille de Zāhir Ibn Rustam al-Isfahānī. Elle était apparemment le cœur de son divan de ġazal intitulé Tarğumān al-Ašwāq. Quand il parle d’elle dans l’introduction de son commentaire personnel de son propre divan, il dit que c’est une fille douce et attirante, qui pare les assemblées et qui fait partie des adoratrices savantes et ascètes. Il ajoute que chaque nom utilisé est une allusion à elle, remarquant aussi que tout ce qu’il a composé l’a été dans la voie de Dieu, toujours selon la méthode soufie 729 : 727 Ibid., p. 175 728 Īmān Abū Duhaym, Ḥiğāziyyāt al-Šarīf al-Raḍī wa Nağdiyyātihi, op. cit., p. 104. 729 Muḥyī al-Dīn Ibn ˁAlī Ibn al-ˁArabī, Tarğumān al-Ašwāq, op. cit., p. 22-24. 288
« Tout nom que je mentionne dans ce recueil fait allusion à elle. Toute demeure dont je fais l’éloge nostalgique est la sienne. Dans cette composition poétique, je n’ai eu de cesse de suggérer les Événements divins, les Réalités spirituelles qui descendent d’elles-mêmes et les correspondances sublimes qui se présentent, selon une méthode allégorique qui est familière de notre Voie. (...) La science qu’elle possède est celle à laquelle je ferai allusion, et nul ne t’informera mieux que quelqu’un de parfaitement instruit (Coran 33/4) (...). J’ai donc exprimé tout cela dans le langage de la poésie amoureuse et celui des vers galants, afin que les âmes s’éprennent de ces modes d’expression et que les amateurs puissent apprécier leur audition. Ce style est bien celui qui convient au lettré distingué, spirituel et subtil 730. » Niẓām prend une telle place dans la poésie d’Ibn ˁArabī qu’une relation est faite entre l’amour pour Niẓām et l’amour pour Dieu. « Lorsque Ibn ’Arabî explicite une allusion à la jeune fille Nezâm comme étant, selon ses propres termes, une allusion à “une Sagesse (Sophia) sublime et divine, essentielle et sacrosainte, qui se manifesta visiblement à l’auteur de ces poèmes, avec une telle douceur qu’elle engendra en lui joie et allégresse, émotion et ravissement”, nous sommes témoins de la transfiguration d’un être que l’Imagination perçoit directement à la hauteur d’un symbole, en l’adossant à une lumière théophanique, c’est-à-dire à une lumière qui en révèle la dimension en au-delà. Dès l’origine et d’emblée, la figure de la jeune fille a été perçue par l’Imagination active au plan visionnaire, où elle se manifeste comme une Figure d’apparition (sûrat mithâlîya) de la Sophia Aeterna 731. » Ibn ˁArabī est différent d’Ibn al-Fāriḍ dans le sens où, chez ce dernier, il n’y avait pas d’histoire d’amour : Ibn al-Fāriḍ était un poète soufi avec une expérience soufie. Mais Ibn al-Fāriḍ était plus proche de Šarīf al-Raḍī qu’Ibn ˁArabī, même si celui-ci a lui aussi cité beaucoup de lieux du Ḥi āz, surtout ceux cités par Šarīf, comme Ḥağir, Rāma, Mina, Ğamarāt, Zamzam, Abraqayn, Ḏī Salam, al-Ḥimā, ˁĀliğ, Laˁlaˁ et Naqā : 730 Ibn ‘Arabî, Maurice Gloton (trad.), L’interprète des désirs, op. cit., p. 23. 731 Henry Corbin, L’Imagination créatrice dans le soufisme d’Ibn ‘Arabi, op. cit., p. 156-157. 289
« Combien tu fais le ġazal à Naqā de Ḥāğir Ô descendant de l’Arabe bédouin Je suis certes arabe et c’est pour cela Que j’adore les blanches et je désire les Arabes 732. » Au début d’un de ses poèmes, il demande à ses amis de se tourner vers al-Kaṯīb qui se trouve à Laˁlaˁ, entre Bassorah et Koufa. Mais, dans son commentaire, il donne un sens à Laˁlaˁ qui est celui d’un état d’étonnement et de désir 733. Puis dans les vers suivants, il parle de son amour : « Ô mes deux intimes, détournez votre chemin En passant par la Dune! Chevauchez vos montures jusqu’à la Halte de Laˁlaˁ Et aspirez aux eaux de Yalamlam Près d’elles, ceux que tu as connus ; Et ceux à qui appartiennent Mon jeûne, mon pèlerinage, ma visite Et ma fête solennelle aux lieux saints. Que jamais je n’oublie le jour où, à Minâ, Les cailloux sont lancés, ni les choses d’importance, Près du suprême autel sacrificiel, Ni près de la source de Zamzam. 732 Muḥyī al-
D
īn Ib
n
ˁ
Alī Ibn al-ˁArabī,
Tar
ğ
umā
n al
-
Ašwāq, op
.
cit., p. 156, v. 27-28 : « kam-tunāġī bi-n-naqā min-ḥāğirin yā-salīla l-ˁarabiyyi l-ˁurubā ana il
lā
ˁarabiyyun wa-li
ḏā
aˁšaqu
l-bīḍa wa-ahwa-l-ˁarabā
. » 733 Ibid, p. 35, v
.
1-4
: « ḫalīlayya ˁūğā bi-l-kaṯībi wa-ˁarriğā ˁalā laˁlaˁin wa-ṭlubā miyāha yalamlamī fi-inna bi- hā man qad-ˁalimta, wa-man lahum fa-lā ansa yawman bi-l-muḥaṣṣabi min minan muḥaṣṣabu-hum qalbī li-ramyi ğimāri-him ṣiyāmī wa-ḥağğī wa-ˁtimārī wa-mawsimī wa-bi-l-manḥari l-aˁlā umūran wa-zamzamī wa-manḥar-hum nafsī wa-mašrabu-hum damī. » 290 Là où ils lancent les pierres Demeure mon cœur, lancé contre leurs pierres. Mon âme, là où ils sacrifient Mon sang, là où ils s’abreuvent. Ô chantre conducteur de chameaux! Si tu viens à Ḥâjir, Arrête un moment les montures Et transmets le salut 734! » Ibn ˁArabī développe ainsi une image de Šarīf al-Raḍī quand il dit, dans une de ses ḥiğāziyyāt : « Ô chamelier des chameaux! Si tu viens à Ḥāğir Arrête les montures un moment, puis transmets les salutations 735. » Et dans plusieurs vers, Ibn ˁArabī parle d’une gazelle qui broute à l’intérieur de lui-même, comme dans le vers suivant : « Que mon père et moi soyons des rançons pour une belle gazelle Que cette gazelle broute entre mes côtes en sécurité 736. » Puis ce poète soufi déclare aussi que son cœur est une prairie à laquelle les gazelles peuvent avoir accès, insistant sur l’image du pâturage qui se trouverait entre les côtes et les entrailles : « Quoi de plus surprenant Qu’une gazelle voilée Montrant un jujubier, Et faisant signe de ses paupières! 734 Ibn ‘Arabî, Maurice Gloton (trad.), L’interprète des désirs, op. cit., p. 88-89. 735 Muḥyī al-Dīn Ibn ˁAlī Ibn al-ˁArabī, Tarğumān al-Ašwāq, op. cit., p. 37, v. 5 : « fa-yā ḥādiya l-ağmāli in-ğiˀta ḥāğiran 736 Ibid., p. 102, v. 6 : « bi-abī ṯumma-bī ġazālun rabībun fa-qif bi-l-maṭāyā sāˁatan ṯumma sallimī. » yartaˁī bayna aḍluˁī fī-amānī. » 291 Une gazelle dont le pâturage Se trouve entre côtes et entrailles! Ah quel prodige! Un jardin au milieu de feux! Mon c œur est devenu capable D’accueillir toute forme. Il est pâturage pour gazelles Et abbaye pour moines 737! » L’image de la gazelle du saule qui broute du cœur du poète est une image que nous avons déjà largement vu chez Šarīf al-Raḍī, dans le vers suivant que nous avons déjà cité plus haut : « Ô gazelle du saule qui broute dans sa charmille Profite aujourd’hui, car le cœur est ta prairie 738. » Commentant cette image du pâturage pour les gazelles qui se trouverait entre les côtes, c’est-à-dire au niveau de la poitrine, Ibn ˁArabī explique : « Le pâturage de cette gazelle se trouve entre les côtes (tarâib) : cet élément d’hémistiche fait allusion aux sciences demeurant dans la poitrine de l’amant, et les entrailles (ḥashâ) désignent les sagesses et la foi qui emplissent (ḥashâ) son intérieur et son cœur, ainsi que [‘Ali, cousin, gendre du Prophète et quatrième calife] l’a exprimé en frappant sa poitrine de la main : “Il y a ici des sciences en abondance! Si seulement je pouvais trouver des personnes pour les recevoir!” Ensuite, il est fait allusion à l’étonnement de l’amoureux qui brûle des feux de l’amour et du désir fervent. Comment, en effet, les sagesses 737 Ibn ‘Arabî, Maurice Gloton (trad.), L’interprète des désirs, op. cit., p. 146-147 : « wa-min ˁağabi l-ašyāˀi ẓabyun mubarqaˁun Wa-marˁā-hu mā-bayna t-tarāˀibi wa-l-ḥašā yušīru bi-ˁunnābin wa-yūši bi-ˀağfānī wa-yā ˁağaban min-rawḍatin waṣṭa nīrānī Laqad ṣāra qalbī qā
bilan kulla ṣūratin 738 Dīwān, T. 2, p. 99, v. 1 (basīṭ) : fa-marˁan li-ġizlānin wa-dayrun li-ruhbānī. » « yā ẓabyata l-bāni tarˁā fī-ḫamāˀili-hi liyahnaki l-yawma anna l-qalba marˁā-kī. » 292 et les sciences qui sont dans les côtes supérieures ou poitrine (tarâ’ib) et les entrailles ne sont-elles pas consumées 739? » Cette dernière poésie d’Ibn ˁArabī ne laisse pas insensible le lecteur qui connaît les écrits de Šarīf al-Raḍī. En effet, en plus de cette image du cœur-pâturage, qui ressemble à un calque de l’image citée plus tôt d’al-Raḍī, cette poésie d’Ibn ˁArabī laisse transparaître de nombreuses similitudes avec les ḥiğ
ā
ziyyāt de notre poète. 740 Ibid., p. 145. 741 Dīwān, T. 2, p. 416, v. 1 (basīṭ) : « yā ṭāˀira l-bāni ġirrīdan ˁalā fanani mā hāğa nawḥu-ka-lī yā ṭāˀira l-bāni. » 293
Avec elle je
converse
Au cré
puscule et
à
l’aube, Rempli d’un désir de tendresse Et d’un amour éploré 742. » On
retrouve
là aussi
, d’une certaine façon, l’ambivalence de Šarīf al-Raḍī, son amour n’allant pas sans le chagrin, la tristesse. Chez ces deux poètes, une forme de souffrance ou de tristesse semble inévitable comme conséquence de l’amour. Et cette passion d’Ibn ˁArabī, il en parle en énonçant plusieurs lieux très caractéristiques des ḥiğāziyyāti d’alRaḍī : « Avec cruels désirs et intense passion, Avec des épreuves nouvelles, Ils viennent jusqu’à moi En prenant de multiples formes. Qui sera à moi à Jam‘ À al-Muḥaççab près de Minâ? Qui sera à moi à Dhât al-Athl? Qui aussi à Na‘mân 743? » En quelques mots, Ibn ˁArabī passe d’un lieu à l’autre et n’énonce pas moins de cinq toponymes, rappelant immanquablement l’amour de Šarīf al-Raḍī pour ces mêmes lieux, ce dernier n’ayant cessé de répéter ces noms-là, et tout particulièrement Mina, avec ses vingt-huit occurrences dans les ḥiğāziyyāt de Šarīf al-Raḍī. Dans une autre poésie, Ibn ˁArabī commence par citer deux noms de lieux : « À Dhû Salam et au monastère, À côté de celui qui est présent à al-Ḥimâ, Des gazelles te laissent voir le soleil, Sous forme de statues de marbre 744. » 742 Ibn ‘Arabî, Maurice Gloton (trad.), L’interprète des désirs, op. cit., p. 145. 743 Ibid., p. 145-146. 744 Ibid., p. 160. 294
Il cite encore plusieurs noms de lieux caractéristiques des ḥiğāziyyāt dans les vers suivants, dans une autre poésie où les femmes et la beauté sont largement mentionnées, des femmes avenantes qui se pressent vers le poète lorsqu’il va « baiser la Pierre noire » : « Ô combien d’âmes fières Avons-nous abattues à Minâ, Près des lieux où les pierres sont lancées Contre les stèles de l’Ennemi Dans Sarḥat al-Wâdi Et les monts de Râma et de Jam‘, Quand les pèlerins déferlent Des monts de ‘Arafât 745. » Ces vers n’étant que quelques exemples parmi d’autres, nous remarquons que la citation de noms de lieux propres aux ḥiğāziyyāt n’est pas anecdotique chez Ibn ˁArabī, qui les cite et les répète, à la manière d’un Šarīf al-Raḍī qui en a fait sa spécificité, sans pour autant garder ˁArafāt secret, ce lieu étant expressément nommé dans le dernier vers susmentionné. Et à la fin de cette même poésie, Ibn ˁArabī termine ainsi en parlant de « femmes parfumées » : « Quand la crainte les prend Elles laissent flotter leurs tresses. Les longues boucles alors paraissent Les draper de ténèbres 746. » Cette dernière image rappelle également l’image vue précédemment chez Šarīf al-Raḍī dans une de ses ḥiğāziyyāt : « Celles qui partirent le soir enveloppées Dans les voiles de la nuit 747. » 745 Ibid., p. 122. 746 I
bid
., p. 123. 747 Dīwān,
T
. 1,
p
.
208
,
v
. 4 (mağzūˀu r-ramal) : rāˀiḥātin fī ğalābībi d-duğā mu tamirāti. 295 Ibn ˁArabī, dans son commentaire, offre des significations symboliques à des lieux géographiques, l’Irak symbolisant la racine et le Yémen la foi, la sagesse, le souffle du Miséricordieux et la douceur du cœur. Ibn ˁArabī, de manière générale, en s’inspirant de Šarīf, joue beaucoup avec les mots, et il aime mettre côte à côte des mots avec la même racine (ğinās), plaçant un toponyme à côté d’un mot proche, comme Mina avec munā (espoir), ˁArafāt avec ˁariftu (j’ai connu), Ğamˁ avec jamaˁnā (nous avons rassemblé). Enfin, dans un autre vers, Ibn ˁArabī dit : « Le Šarīf, c’est le pieux dont on est satisfait (al-Murtaḍā) 748. » Ce vers semble être un clin d’œil pour montrer sa proximité avec la famille de Šarīf alRaḍī de manière générale, ses propos faisant directement penser à son frère aîné Šarīf alMurtaḍā. Un respect spécifique est d’ailleurs accordé au titre de Šarīf que porte al-Raḍī, du fait qu’il soit un descendant du Prophète de l’Islam. Ces quelques mots peuvent d’ailleurs porter plusieurs sens, « al-Murtaḍā » pouvant être compris dans son sens littéral, ou bien renvoyer au nom que l’on connaît, et ainsi, l’adjectif « pieux » ne ferait qu’accroître le respect donné à un tel personnage, puisque le vers soutiendrait qu’alMurtaḍā est pieux, d’une part, et qu’il est le meilleur représentant de la position de Šarīf, d’autre part. Les ḥiğāziyyāt ont donc fait leur entrée dans la poésie soufie. En outre, certains ont profité des ḥiğāziyyāt pour faire l’éloge du Prophète (madḥ al-rasūl), même si ces éloges existaient depuis l’époque même du Prophète, sur le modèle de Ḥassān Ibn ābit et Kaˁb Ibn Zuhayr. Toutefois, ce que nous voyons par la suite dans les nouvelles poésies, comme avec Ṣarṣarī (m. 656h/1258) et al-Buṣayrī (m. 696h/1297), qui ont eu un rôle dans la cristallisation des madāˀiḥ (éloges du Prophète), c’est la diffusion des ḥiğāziyyāt, particulièrement chez les soufis, qui ont accordé une très grande place aux lieux du Ḥiğāz. Le madīḥ classique existait, et le genre des ḥiğāziyyāt a réussi à s’y intégrer. Même Ismaˁīl al-Nabhānī (m. 1350h/1931), juge palestinien qui a vécu une partie de sa vie à Médine, qui a collecté tous les éloges du Prophète depuis Kaˁb Ibn Zuhayr jusqu’à ses propres 748 Muḥyī al-Dīn Ibn ˁAlī Ibn al-ˁArabī, Dīwān Ibn ˁArabī, op. cit., p. 58, v. 2 : « inna š-šarīfa huwa t-taqiyyu l-murtaḍā. » 296 poésies de madāˀiḥ (vingt-cinq mille soixante-neuf vers tirés de deux cent treize poètes), note : « Il est recommandé pour celui qui fait l’éloge du Prophète qu’il montre son désir pour les lieux du Ḥiğāz et ses traces et l’amour de cette région, le désir tourné vers eux et les pleurs, et qu’il fasse la description des chameaux, du voyage et des lieux d’abreuvement, qu’il décrive les nuages, les éclairs et les vents qui viennent de leur côté, qu’il fasse des invocations pour les gens du Ḥiğāz et leurs foyers 749. » Il rapporte aussi, de Taqiyy al-Dīn Ibn Ḥuğğa al-Ḥanafī (m. 837h/1434), dans son livre Ḫizānat al-Adab : « Le ġazal avec lequel on commence le prologue de l’éloge du Prophète, le poète doit être poli et modeste, et il doit chanter des poésies d’amour en citant Salˁ, Rāma, l’avant-mont de ˁAqīq, al-ˁUḏayb, al-Ġuwayr, Laˁlaˁ et les côtés de Ḥāğir 750 » Ainsi, il était devenu normal et logique de citer les lieux des ḥiğāziyyāt dans toute volonté poétique de faire l’éloge du Prophète. Al-Šībī remarque que les poètes de madāˀiḥ ont bien profité des ḥiğāziyyāt et des lieux du Ḥiğāz, surtout de Médine, terre du Prophète, et que les soufis se sont accrochés aux ḥiğāziyyāt en tant que pays de leur exemple suprême 751. Nous remarquons par exemple que le poète Ṣarṣarī al-Baġdādī (m. 656h/1258), du village de Ṣarṣar près de Bagdad, un des fondateurs des éloges du Prophète, tué par les Tatars en défendant 749 Muṣṭafā al-Šībī, « Hiğāziyyāt al-Šarīf al-Raḍī », Al-Šarīf al-Raḍī Dirāsāt fī Ḏikrāh al-Alfiyya, op. cit., p. 54 : « wa-yustaḥsanu li-man madaḥa n-nabiyya an-yušabbiba bi-ḏikri d-diyāri l-ḥiğāziyyati wa-maˁālimi-hā wa-ḥubbi sukkāni-hā wa-š-šawqi ilay-him wa-l-bukāˀu wa-waṣfu n-niyāqi wa-s-sayri wa-l-manāhili wawaṣfu s-saḥābi wa-l-barqi wa-r-rīḥi l-latī tağīˀu min-naḥwi-him wa-d-duˁāˀu la-hum bi-s-salāmati wa-dduˁāˀu la-hum bi-s-salāmati wa-li-diyāri-him. » 750 Ibid : « inna l-ġazala l-laḏī yuṣaddaru bi-hi l-madīḥu n-nabawiyyu yataˁayyanu ˁalā n-nāẓimi an-yaḥtašima fī-hi wa-yataˀaddaba wa-yataḍāˀala wa-yatašabbaba muṭraban bi-ḏi
salˁin wa-rāmata wa-safḥi l-ˁaqīqi wal-ˁuḏaybi wa-l-ġuwayri wa-laˁlaˁin wa-aknāfi ḥāğirin. » 751 Ibid, p. 52. 297 Bagdad, commence ses vers par la nostalgie et le désir qu’il porte pour les lieux du Ḥiğāz et des lieux sacrés : « Ô larmes des yeux, ton rendez-vous est la séparation Tu as mis en réserve du sang qui coule Que veut dire le fait que le passionné fasse attention le jour de la séparation À ses larmes, quand les amis sont partis? Ô caravane du Ḥiğāz, que tu sois guidé pour que tu aies pitié Avec un cœur passionné qui avance avec vous Je suis surpris de voir qu’il descend à Ḏāt ˁIrqayn Avec tous ses efforts, alors que sa maison est en Irak Et il réside sur la terre de Naˁmān avec désir Et les chameaux n’ont pas senti sa marche Ô nuits de Ḫayf de Mina, est-ce que se font Des buts sous votre ombre ou est-ce un obstacle? Ô terre de la plaine de la Mecque! Y a-t-il un moyen Pour une rencontre dont jouit le groupe? » Comme Šarīf, ensuite, il revient vers Médine quand il parle de Salˁ : « Y a-t-il un retour possible aux montagnes de Salˁ Pour un amoureux qui est toujours passionné 752? » 752 Īmān Abū Duhaym, Ḥiğāziyyāt al-Šarīf al-Raḍī wa Nağdiyyātihi, op. cit., p. 106 : « dumūˁa l-ˁayni mawˁidu-ki l-firāqu hunālika mā-ḫazanti daman yurāqū wa-mā rifqu l-mutayyami yawma baynin ayā-rakba l-ḥiğāzi hudīta rifqan bi-admuˁi-hi wa-qad sāra r-rifāqū bi-qalbin hāˀimin maˁa-kum yusāqū ˁağibtu la-hu yaḥill
u bi-ḏāti ˁirqin wa-yaskunu arḍa naˁmāna štiyāqan bi-himmati-hi wa-manzilu-hu l-ˁirāqū wa-lam tašˁur bi-masrā-hu n-niyāqū fa-yā-laylāti ḫayfi minan atuqḍā wa-yā-baṭḥāˀa makkata hal-sabīlun maˀāribu fī-ẓilāliki am-tuˁāqū ilā-waṣlin yaliḏḏu bi-hi l-maḏāqū [...] 298
Dans cette poésie, nous retrouvons les sens et l’âme de Šarīf, avec les nuits de Ḫayf de Mina, la terre de Naˁmān, la Mecque, le retour vers Salˁ à Médine, son cœur partagé entre l’Irak et le Ḥiğāz. Tout cela nous rappelle clairement les ḥiğāziyyāt d’al-Raḍī. Après Ṣarṣarī, au Maroc, un autre grand poète de madāˀiḥ a écrit ce qui est peutêtre la poésie la plus connue d’éloges du Prophète. Son auteur, al-Buṣayrī (m. 696h/1297), raconte : « Plus tard, je fus atteint d’un mal terrible qui paralysa la moitié de mon corps et m’empêchait de bouger. Me vint alors l’idée de composer une œuvre qui traitait des mérites du Prophète, à travers laquelle j’invoquai Dieu de me guérir. Après l’avoir composée et m’être endormi, voilà que m’apparut en songe le Prophète. Je lui récitai mon œuvre en entier, et il caressa de sa noble main bénie les parties paralysées du corps de l’humble et ingrat serviteur que je suis. À mon réveil, j’étais complètement rétabli et n’éprouvais plus aucune douleur 753. » Il commence son poème par le vers suivant : « Est-ce le souvenir de tes voisins (bienheureux) de Ḏū Salam qui fait jaillir de tes prunelles ce flot de larmes mêlées de sang? ou est-ce en raison du vent impétueux qui souffle de Kāẓima, ou bien des éclairs qui fendent les ténèbres de Iḍam 754? » Le poète Al-Raḍī, avant lui, écrit, avec le même mètre basīṭ que nous voyons dans la poésie susmentionnée : « Si ce n’était le souvenir de ces jours à Ḏī Salam 755. » wa-hal raddun ilā aˁlāmi salˁin li-ṣabbin lā-yuzāyilu-hu štiyāqū. » 753 Sharaf al-Dîn Muhammad al-Sanhâjî, Hassan Boutaleb (trad.), Al-Burda : La perle des odes et le diadème des hymnes
à la gloire du Prophète, op. cit., p. 15. 754 Ibid., p. 36.
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Conflicts of Interest: The author declares no conflict of interest. Conflicts of Interest: The author declares no conflict of interest.
Funding: This research received no external funding. References 1. Browse All Publishers. SciLit. Available online: https://www.scilit.net/statistic-publishers
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online: https://scholarlykitchen.sspnet.org/2020/08/10/guest-post-mdpis-remarkable-growth/
(accessed on 23 July 2021). (
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3. Celebrating 25 Years of MDPI during 2021. Available online: https://www.mdpi.com/
anniversary25 (accessed on 22 July 2021). Publisher’s Note: MDPI stays neutral
with regard to jurisdictional claims in
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iations. Editorial
Publisher’s Note: Rheumato—A New Open Access Journal Clàudia Aunós MDPI, Avinguda Madrid, 95, 5-3, 08011 Barcelona, Spain; claudia.aunos@mdpi.com; Tel.: +34-93-639-7662 As one of the top four scholarly journal publishers in the world [1,2], MDPI is
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Citation: Aunós, C. Publisher’s Note:
Rheumato—A New Open Access
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Received: 28 July 2021
Accepted: 5 August 2021
Published: 12 August 2021
Citation: Aunós, C. Publisher’s Note:
Rheumato—A New Open Access
Journal. Rheumato 2021, 1, 1. https://
doi.org/10.3390/rheumato1010001
Received: 28 July 2021
Accepted: 5 August 2021
Published: 12 August 2021 Publisher’s Note: MDPI stays neutral y
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jpm (accessed on 23 July 2021). 5. Journal of Cardiovascular Development and Diseases Home Page. Available online: https://www. mdpi.com/journal/jcdd (accessed on 23 July 2021). 6
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Available online: https://www mdpi com/journal/tomography 5. Journal of Cardiovascular Development and Diseases Home Page. Available online: https://www. mdpi.com/journal/jcdd (accessed on 23 July 2021). 6
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Interwar Blueprints of Europe: Emotions, Experience and Expectation
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www.ssoar.info Issue Issue
This article is part of the issue “Interdisciplinary Approaches to Studying Emotions within Politics and International Rela-
tions”, edited by Alex Prior (University of Leeds, UK) and Yuri van Hoef (Utrecht University, The Netherlands). © 2018 by the author; licensee Cogitatio (Lisbon, Portugal). This article is licensed under a Creative Commons Attribu-
tion 4.0 International License (CC BY). There is but one way to avert that ruin: the economic
federation of Europe’s continental democracies….This is
the only road to salvation, alike for European politics
and European industry. (Coudenhove-Kalergi,
1923, p. 70, author’s own translation) This article aims to further expand on this by ex-
plicitly focusing on the role of emotional vocabulary in
relation to the meaning of key concepts of European
unity. To this end, it analyzes a canonical text of the time,
with a transnational outreach: Coudenhove-Kalergi’s
Pan-Europa (1923). Trineke Palm Submitted: 3 April 2018 | Accepted: 21 August 2018 | Published: 28 December 2018 Submitted: 3 April 2018 | Accepted: 21 August 2018 | Published: 28 December 2018 Abstract The notion of European integration has been contested from its very start. In the interwar period many ideas were floating
around on how to shape European unity. These interwar Blueprints for Europe have to be understood in the context of
conflicting and contradictory emotions of enmity and amity. This article looks at the emotive vocabulary of the canonical
text of Coudenhove-Kalergi’s Pan-Europa. It applies an emotion discourse analysis, using Koselleck’s notion of “space of
experience” and “horizon of expectation”. As such it shows the connection between the understanding and use of time
and emotions in discourse—thereby demonstrating the necessity of “reading” the blueprints of European integration as
highly normative and moral claims on the design of this European order. Keywords Keywords
discourse; Coudenhove-Kalergi; European integration; emotion; interwar; pan-Europa Politics and Governance, 2018, Volume 6, Issue 4, Pages 135–143 www.ssoar.info Empfohlene Zitierung / Suggested Citation:
Palm, T. (2018). Interwar Blueprints of Europe: Emotions, Experience and Expectation. Politics and Governance, 6(4),
135-143. https://doi.org/10.17645/pag.v6i4.1522 Empfohlene Zitierung / Suggested Citation:
Palm, T. (2018). Interwar Blueprints of Europe: Emotions, Experience and Expectation. Politics and Governance, 6(4),
135-143. https://doi.org/10.17645/pag.v6i4.1522 Terms of use:
This document is made available under a CC BY Licence
(Attribution). For more Information see:
https://creativecommons.org/licenses/by/4.0 Nutzungsbedingungen:
Dieser Text wird unter einer CC BY Lizenz (Namensnennung) zur
Verfügung gestellt. Nähere Auskünfte zu den CC-Lizenzen finden
Sie hier: https://creativecommons.org/licenses/by/4.0/deed.de Politics and Governance (ISSN: 2183–2463)
2018, Volume 6, Issue 4, Pages 135–143
DOI: 10.17645/pag.v6i4.1522 Interwar Blueprints of Europe: Emotions, Experience and Expectation Trineke Palm 2.1. Ideas, Emotives and Emotional Beliefs The emotional turn in IR builds on research in the history
and politics of emotions. Frevert’s (2011) work on the his-
tory of emotions is well-known, showing that our under-
standing of emotions has changed over time. Moreover,
in The Cultural Politics of Emotion, Ahmed (2014) aims
to show how the emotionality of text connects individu-
als to a collective, which includes some at the expense
of others. However, little is known of the ways in which these
substantive ideas for a future European order were con-
nected with specific emotives and emotional beliefs. Hence, building upon the “emotional turn” in interna-
tional relations (IR), this article develops a framework of
analysis that highlights the role of emotives and emo-
tional beliefs that were evoked in these transnational
visions of Europe, and that served to create moral and
political support for them. In particular, it aims to show
how experience and expectation are connected by emo-
tives. As such, the focus on how emotives are used to
strengthen the substantive ideas (moral/spiritual, polit-
ical and economic) gives us new insights on how a fu-
ture Europe was invoked and made attractive to a wider
public/audience. Research on emotions in IR consists of both macro-
theoretical
approaches
that
develop
generalizable
propositions about political emotions, and micro-
approaches that examine specific emotions in specific
contexts, often decision-making failures (Hutchison &
Bleiker 2014; see also Sasley, 2011). The “emotional
turn” in IR encompasses a wide variety of approaches
and concepts, ranging from a focus on individual sub-
jective experiences (emotions) and bodily expressions
(affection) to more collective systems of feeling (emo-
tional communities, Rosenwein, 2010; Koschut, 2018a),
emotional cultures (Moïsi, 2009), institutionalized emo-
tional norms (emotional regimes, Reddy, 2001; see also
Crawford, 2000) and conceptual/abstract expressions
(emotional beliefs, Mercer, 2010). Moreover, beyond the topic of European integration,
this article aims to advance research on emotions in IR. While the literature remained rather theoretical or fo-
cused on single emotions to account for decision-making
failures (Sasley, 2011; see also Bleiker & Hutchinson,
2008), the methodology of studying emotions in (the his-
tory) of IR gained more attention recently (see Clément &
Sangar, 2018). Building upon Koschut (2018b), I develop
an emotion discourse analysis, distinguishing between
two types of emotion (enmity and amity), enriched by
combining this with Koselleck’s (2005) notions of “space
of experience” and “horizon of expectation”. 2.1. Ideas, Emotives and Emotional Beliefs As such this
article engages with the need for more structured, sys-
tematic empirical research on emotive vocabulary in Eu-
ropean integration history. This article does not engage with questions about the
biological or cultural basis of emotion, but rather con-
ceives of emotional expressions as a strategy to constrain
or enable particular substantive ideas about Europe. As
such, it does not refer to emotions, but emotives. Emo-
tives are expressed emotions (i.e., emotional vocabulary)
that are instrumental in changing, building, hiding and in-
tensifying emotions that are accepted and recognized in
a particular community (see Reddy, 2001, p. 105). Emo-
tions are inadmissibly present in any political discourse,
however much this discourse tries to present its lan-
guage in impartial, objective and rational means. In fact,
doing so is in itself an emotional strategy, in the sense
that it tries to channel specific “non-emotional” emo-
tions, in order to render its contents more convincing. The central aim of this article is to examine how par-
ticular emotives and emotional beliefs were used to gain
support for a specific and salient blueprint of European
integration. To what extent and how are the experiences
and expectations, as expressed in Coudenhove-Kalergi’s
Pan-Europa, informed by emotives? Blueprints are understood here as a set of moral/
spiritual, economic and political ideas about a future Eu-
ropean order, that are expressed in a particular emotive
vocabulary. The next section discusses the relationship
between ideas, emotives and emotional beliefs in gen-
eral, and emotion discourse analysis in particular. Sub-
sequently, I introduce Coudenhove-Kalergi’s Pan-Europa
(1923) as a canonical text, and emotional landmark,
of interwar writings on European unity. In Sections 4
and 5 I analyze the emotive vocabulary of Coudenhove-
Kalergi’s text, focusing on the “space of experience” he
refers to and his outline of Europe’s future (“horizon of Of equal importance in the analysis are emotional be-
liefs (Mercer, 2010). In contrast to emotives that refer
to basic emotions, emotional beliefs are more abstract
concepts that integrate emotion and cognition (Mercer,
2010, p. 2). Examples of emotional beliefs, in the context
of this article, are nationalism, responsibility, solidarity
and reconciliation. Emotion is not an addition to these
concepts, but is an essential component for understand-
ing its meaning (Mercer, 2010, p. 7). 1. Introduction There is ample research on the postwar process of
European integration based on a rationalist account of
European integration as the outcome of the autonomous
calculations of different nation states based on their
economic, political or military interests (e.g., Milward,
1992), and ideational accounts focusing on transnational
networks, their norms and institutions, that pushed
for European integration (e.g., Kaiser, 2007; Lipgens,
1968; Müller, 2004). While Lipgens’ (1968) work on the
blueprints of national resistance movements has been
criticized for its “normative overdrive” (Kaiser, 2007),
it shows that there is a wide variety of blueprints of
Europe that transcend particular national views. While
these networks did not necessarily produce a common
political European idea, they provided a forum for col-
lective learning, and fostered mutual understanding and
trust (Kaiser, 2004). Moreover, they created normative- Before the European integration project started to insti-
tutionalize into the European Coal and Steel Community
(ECSC) in 1951, many ideas floated around on how to
shape European integration. Already during the interwar-
period and during World War II (WWII) there were sev-
eral plans that envisioned a united Europe. These inter-
war years were the “first golden age” of the European
ideal among intellectuals (Lacroix & Nicolaïdis, 2010). Re-
cently, Cattani (2017) combined conceptual history with
discourse analysis to examine the meaning of concepts
such as “European homeland” and “European nation”
in the context of the two intellectual conferences orga-
nized by the League of Nations in 1933. Similarly, Ifversen
(2002) analyzed the interconnected notions of crisis, civ-
ilization and Europe in the interwar era. Politics and Governance, 2018, Volume 6, Issue 4, Pages 135–143 135 expectation”). In Section 6, based on secondary litera-
ture, I reflect upon the emotionalizing effect, reception
and response. emotional bonds able to withstand domestic/nationalist
pressures (Kaiser, 2007). Concerning the interwar-period, numerous transna-
tional networks can be identified that did contribute to
these normative-emotional bonds, such as the Roman-
Catholic networks (Secrétariat International des Par-
tis Démocratiques d’Inspiration Chrétienne), ecumenical
networks (Life & Work, Faith & Order) and business net-
works (Council of Directors of European Industrial Feder-
ations [CDEIF]; Colpach-Group). Politics and Governance, 2018, Volume 6, Issue 4, Pages 135–143 2.2. Emotion Discourse Analysis For the purpose of this article, a canonical text on Eu-
ropean unity in the interwar period has been selected:
Coudenhove-Kalergi Pan-Europa (1923). An emotion discourse analysis looks into the emotion
potential and emotionalizing effect of political discourse
(Koschut, 2018b) and is concerned not so much with
the frequencies of specific emotives, but rather with
the prevalence of certain emotional patterns/structures,
i.e., a certain combination of emotives and emotional
beliefs. These master emotives “structure the social
meanings and effects of the discourse and thus set the
collective standard of emotional expression” (Koschut,
2018b, p. 294). Coudenhove-Kalergi’s writings are among the first of
blueprints of European unity in the interwar-era. Dur-
ing this “first golden age” of intellectual debate on Eu-
ropean unity over 600 articles were published on the
topic between 1919 and 1939, with a peak between 1925
and the early 1930s (Chabot, 2005, p. 14). While the in-
terwar period was characterized by intellectual debate
on European unity, it was also a period of thriving na-
tionalism. In the same year that Pan-Europa was pub-
lished, national socialist party leader Hitler called for a
“national revolution”, in strong emotives: “what moti-
vates us is neither self-conceit or self-interest, but only
a burning desire to join the battle in this grave eleventh
hour for our German Fatherland” (Frevert, 2015). More-
over, Coudenhove-Kalergi’s work has been compared to
Rohan’s Europa (published in 1924 at the same pub-
lisher as Pan-Europa), which proposed a culturally united
Europe built on a shared Catholic basis (Prettenthaler-
Ziegerhofer, 2013, pp. 167, 176). With 16,000 copies
being sold in 1926 Pan-Europa was a best-seller at
its time (Prettenthaler-Ziegerhofer, 2012). Translated in
many languages it also had a clear transnational reach
and was discussed in transnational settings such as the
Pan-European conferences that Coudenhove-Kalergi or-
ganized as part of his Pan-European Union. At the first
conference, in 1926, 2,000 participants from 24 nations
gathered in Vienna. As such, the Pan-European Union
“constituted a public space where both the meaning of This article combines emotion and time in discourse
(see also Skonieczny, 2018, p. 70). Concerning emo-
tive vocabulary, I distinguish between emotions associ-
ated with amity (i.e., empathy, pride, gratitude, grief,
honor, respect, compassion and sympathy) and enmity
(i.e., fear, anger, disgust, hatred, jealousy, rage) (Koschut,
2014). 2.1. Ideas, Emotives and Emotional Beliefs These conceptual-
izations of emotives and emotional beliefs point out that,
although research on emotions is often focused on the
level of the individual, there is also a strong collective 136 Politics and Governance, 2018, Volume 6, Issue 4, Pages 135–143 and political dimension to it: “[Emotions] frame what is
and is not possible in politics. They reveal and conceal,
enable and disable” (Hutchison & Bleiker, 2014, p. 508). Moreover, they contribute to identity formation, possi-
bly resulting in an emotional community. to experience. In contrast, forward looking emotions are
fear and hope. An example of an emotive that connects
experience with expectation is revenge, i.e., it refers to
anger based on the past, with the conviction to make up
for that in the future. These emotives are not an exhaus-
tive list, but rather examples to show the connection be-
tween emotives and notions of time. Moreover, while
analytically distinctive, empirically these emotions may
be intertwined. The connection of past and future, of ex-
perience and expectation, leads to particular blueprints
of Europe. As such, emotions are closely tied to morals in at least
two ways. First, emotional regimes prescribe what emo-
tives are “appropriate”. As such, morals precede emo-
tions. However, emotions and morals also are in a consti-
tutive relationship. So-called moral emotions entail a cer-
tain judgment, condemning others (i.e., contempt, anger
and disgust), or praising others (i.e., gratitude, awe, ele-
vation) (Haidt, 2003). The emotion discourse analysis proceeds in three
steps (Koschut, 2018b). First, the selection of texts (Sec-
tion 3). Second, the mapping of verbal expressions of
emotions (Sections 4 and 5). Third, interpreting and con-
textualizing its political effects (Section 6). It is the connection between ideas and emotives that
serves as a mobilization for action. Without ideas, emo-
tives have not object, and without emotives, ideas fail
“to produce a sense of obligation or loyalty necessary for
collective action” (Koschut, 2018b, p. 286). 2.2. Emotion Discourse Analysis As we will
see, his work consists of emotives with a strong reli-
gious connotation. Coudenhove-Kalergi’s approach is based on a neo-
aristocracy, i.e., a blend of “forward-looking enthusiasm
for technological and industrial progress and a backward-
looking authoritarian elitism” (Sorrels, 2016, p. 137). While Coudenhove-Kalergi opposed Hitler and was un-
equivocal in his condemnation of Nazi anti-Semitism, he
flirted with Mussolini’s fascism, emphasizing the need for
a “wise mixture of the authoritarian and aristocratic prin-
ciple with what can be healthy in the democratic princi-
ple [conceding] room to justice and the enlightened com-
mand of superior personalities” (Evola, 1933/2015, p. 47;
see also Coudenhove-Kalergi , 1934, pp. 94–96). Hence, although contested, his writings have been
central to the interwar thinking about European unity. As such this text is taken as an “emotional landmark”
(Koschut, 2018b).3 As a product of its time, Pan-Europa
is a telling example of the use of emotive vocabulary in
the interwar years. It tells us something about the recog-
nized and accepted emotions among the transnational
European elites of that time. While initially a great admirer of the American Pres-
ident Woodrow Wilson, Coudenhove-Kalergi became
strongly disappointed in Woodrow Wilson’s League of
Nations, which suffered from utopian overstretch and
did too little for the European dream of a unified conti-
nent: “Wilson’s attempt to set up an ecumenical League
of Nations has failed” (Coudenhove-Kalergi, 1923, p. 20). Coudenhove-Kalergi eventually abandoned his support
for the League of Nations in favor of a United States of
Europe, inspired by Fried’s Pan-Amerika (1910)1. In this,
he pitted his plan for Europe squarely against the ‘Red In-
ternational’, channeling both conservative, 19th century
concert ideas and new, fascist dreams for a rejuvenated
European unification. As we will see, he does so by posit-
ing negative emotives of revenge and destruction against
positive emotives of reconciliation and understanding. The United States of Europe is presented as the 6th
Europe, after Hellas, Roman empire, Völkerwanderung,
Roman-Catholic Europe, and the era of enlightened abso-
lutism. This way Coudenhove-Kalergi (1923, pp. 31–33)
presents Pan-Europa as a continuation of a century old
project. This Pan-Europa would consist of 26 states (ex-
cluding England and Russia)2. After a Pan-European con-
ference, the participating countries would sign an obliga-
tory arbitration and guarantee treaty, eventually leading
to a customs union. 2.2. Emotion Discourse Analysis Moreover, building upon Koselleck (2005), I distin-
guish between the “space of experience” and “horizon
of expectation” of this emotional vocabulary. No emo-
tive can be crafted and disseminated without the pre-
supposition of an existing, collective reservoir of experi-
ences and expectations to tap into. “Space of experience”
refers to references to past experiences and reflections
on the state of affairs. “Horizon of expectation” refers
to references to a future European order. Both can be
combined by a positive, inclusive emotional vocabulary
or a more negative, exclusive emotional vocabulary (see
Table 1). So, negative backward emotions are anger and
hatred, whilst trust is a positive emotion with reference Table 1. Coding scheme emotion discourse analysis. Table 1. Coding scheme emotion discourse analysis. Space of Experience
Horizon of Expectation
Enmity
Hatred, Anger
Fear
Revenge
Amity
Trust
Hope
Politics and Governance, 2018, Volume 6, Issue 4, Pages 135–143
137 Space of Experience 137 atively in the EU integration history literature; he was
ineffective (Horner, 2001), and a marginal political fig-
ure (Milward, 1997). His political ideas have been criti-
cized as well: “As a blueprint it was imprecise, naïve and
optimistic” (White, 1989). Nevertheless, Coudenhove-
Kalergi’s movement was well-known and contributed to
a coalition of idealism and commercial interests that
was of importance after WWII (Klemann, 1966). Hon-
orary president of the Pan-Europa Movement and Prime
Minister of France, Aristide Briand, proposed a Euro-
pean Federal Union in 1930 at the League of Nations. Af-
ter the war, with the emergence of many other move-
ments for European integration, Coudenhove-Kalergi’s
position was less central. However, his work was ac-
knowledged, as he was mentioned by Churchill in his fa-
mous speech at the University of Zürich in 1946: “Much
work has been done upon this task by the exertions of
the Pan-European Union, which owes so much to the
famous French patriot and statesman Aristide Briand
(Churchill, 1946).” Moreover, he was awarded the first
Charlemagne Prize in 1950, acknowledging his efforts for
promoting European unity. Europe and the vision of a united Europe were discussed”
and “provoked new conceptualisations of Europe as a
bearer of common identity” (Orluc, 2002, pp. 25, 43). Although Coudenhove-Kalergi, born in 1894, did
have a Catholic background, he “generally ignored
the theology of his time” (Sorrels, 2016, p. 213). His
biographer Prettenthaler-Ziegerhofer (2001) classifies
Coudenhove-Kalergi as a Christian socialist. 1 For an extensive discussion of the relationship between Fried’s work and that of Coudenhove-Kalergi , see Sorrels (2016).
2 England had transformed into “an oceanic empire” with non-European interests, and Russia distanced itself from the democratic system of Europe by
proclaiming Sovietism (Coudenhove-Kalergi, 1923, p. 33).
3 Other relevant writings of Coudenhove-Kalergi in this period consist of Paneuropa ABC (1931), Kampf um Paneuropa (1925) and Europe Must
Unite (1938). 2.2. Emotion Discourse Analysis So, Coudenhove-Kalergi prioritizes
political and security integration over economic integra-
tion, i.e., a customs union is to be developed only after
political treaties and security pacts are agreed upon. 4. Space of Experience in Pan-Europa In this section I will take a closer look at the emotional
vocabulary of Coudenhove-Kalergi concerning Europe’s
past and how he understands this to affect his contempo-
raries’ emotional outlook. First, we identify Coudenhove-
Kalergi’s understanding of emotions in international pol-
itics. Second, we will see how Coudenhove-Kalergi con-
nects the key emotives of his time (hate and fear) with
(a) an emotional belief in nationalism, and (b) a narrow
space of experience. 4.1. Emotions Trump Interests While the role of emotions in inter-state relations is de-
bated (see Bleiker & Hutchison, 2018), in Coudenhove-
Kalergi’s writings they are omnipresent. He argues that
“not only are political relations between states func-
tions of national sympathy and antipathy—but national
friendships and enmities are also often functions of
political relations” (Coudenhove-Kalergi, 1923, p. 122). Coudenhove-Kalergi even claims that the emotional di- The life and work of this Austrian-Japanese politi-
cian and philosopher have been evaluated rather neg- Politics and Governance, 2018, Volume 6, Issue 4, Pages 135–143 138 This progress in the rest of the world contrasted
with Europe’s decline (Coudenhove-Kalergi, 1923, p. VIII)
is due to the fact that the (technical) abilities to ex-
pand the horizon of expectations are not grasped by
his contemporaries—they lack the necessary imagina-
tion (Phantasie) and are backward-looking, according to
Coudenhove-Kalergi (1923, p. 98). Because of a lack of
imagination, Coudenhove-Kalergi sees the European pol-
itics of his days as yesterday’s politics rather than the pol-
itics of tomorrow (1923, p. VIII). Instead of focusing on
how to prevent a future war, more space in the discus-
sions is given to the last war (1923). This experience of
war is a too weak foundation to prevent another one7. mension of the relations among states (Germany and
France in particular) trumps rationality and interests4,
and that these emotions are artificially created over
centuries for political-military purposes (Coudenhove-
Kalergi, 1923, pp. 121–122)—a typical expression of
a lingering 19th century preference for “concert” and
great power understanding and deliberation (De Graaf,
in press). 4 “Never was the hate between Germans and French bigger than it is now. This hatred for each other is stronger than reason, stronger than their common
interest” (Coudenhove-Kalergi, 1923, pp. 121–122, author’s own translation).
5 “Everywhere there is misery, unrest, discontent, hatred and fear” (Coudenhove-Kalergi, 1923, p. VIII, author’s own translation).
6 “These states of the West are disjointed and disorganized, in everlasting conflict with one another, and torn by hatred and jealousy (Coudenhove-Kalergi,
1923, p. 23)”; “controlled by mutual hatred and envy, by revenge and resentment” (Coudenhove-Kalergi, 1923, p. 95).
7 “To base the hope for peace on this foundation is short-sighted and hopeless” (Coudenhove-Kalergi, 1923, p. 104, author’s own translation). 4.2. Hate, Fear and Nationalism The defining space of experience, against which
Coudenhove-Kalergi writes Pan-Europa is World War I
(WWI). Against this backdrop, the master emotives that
Coudenhove-Kalergi identifies are: hate (Hass) and fear
(Furcht)5. These emotives of jealousy, hatred, envy, re-
venge, and malice6 fuel a vicious circle of eternal fight
in which Europe tumbles from one crisis to another. Coudenhove-Kalergi connects these emotives of hate
and jealousy to nationalism and argues that this toxic
combination of national hate and national jealousy re-
sults in a descent, from month to month, of the spiri-
tual and material European culture (Coudenhove-Kalergi,
1923, p. 68). Feelings of hate trump those of solidar-
ity (p. 71). So, in Coudenhove-Kalergi ’s understanding, the eter-
nal focus on “yesterday” is the main cause of Eu-
rope’s fragmentation (1923, p. VIII). Hence, he suggests
the following tactic: Das Übermorgen gegen das Mor-
gen ausspielen [To Set the Day after Tomorrow Against
Tomorrow] (p. 87). In other words, he aims to propose to
widen the horizon of expectation and accuses his oppo-
nents of limiting that same horizon and only looking back. 5.1. Anti-Europe: Europe of the Past—Revenge &
Destruction The default option is to continue on the road of hate,
which would inevitably cause Europe’s demise, con-
quered by the upcoming powers (i.e., the United States
and Russia) (1923, pp. 21–22). This default option is
based on a vicious circle of French politics of destruc-
tion and a German politics of revenge. Unforgiveness
on both sides lead to a politics of respectively destruc-
tion and revenge (p. 134). These two are closely inter-
linked, and Coudenhove-Kalergi is very much aware of
the strength of revenge politics: it is fueled by every act
of violence (p. 131). 4.3. A Narrow Space of Experience Coudenhove-Kalergi (1923) observes that the space of
experience expands. Because of technical progress, the
world becomes smaller (p. 18). Hence, in terms of the
“space” of these emotives, Coudenhove-Kalergi (1923)
notes that the effects of these feelings of hate are
not restricted to the European continent but contami-
nate the international atmosphere (p. 23). He refers to
Europe as an eternal source of uncertainty and anxiety,
with analogy to the tempestuous developments in the
Balkans (p. 24). Moreover, taking an outside-in perspec-
tive, Coudenhove-Kalergi (1923) portrays an image of
Europe as once being “feared”, but now “pitied” and de-
rided by the upcoming powers (pp. 17, 27). 5. Horizon of Expectation in Pan-Europa In this section we will look at the use of emotional vo-
cabulary by Coudenhove-Kalergi regarding his proposed
future European order. He distinguishes between two
options—a third option does not exist (1923, p. 54). The
fight between anti-European and pan-European is a fight
between “Vergangenheit und Zukunft” (p. 168). Key to
this choice for a Europe of the past, or a Europe of the
future, is the Franco-German relationship. As such, it is the “national chauvinist”, who in prop-
agating a European war shows that blind hate against
the enemy is stronger than love of one’s own (p. 101),
that is Coudenhove-Kalergi ’s (1923) main antagonist. On
Coudenhove-Kalergi ’s (1923) list of opponents, they are
followed by the communists, militarists and protected in-
dustries (p. 162). Russia is seen as merely taking advan-
tage of Europe’s internal weakness. Coudenhove-Kalergi
(1923) even seemed to resist too much of an Ameri-
can interference in European matters: “neither the West
nor the East will save Europe: Russia wants to conquer
Europe, America wants to buy it” (p. XI). 8 Other interwar intellectuals referring to Europe as maternal at symposia initiated by the League of Nations in 1933 in Paris and Madrid are the French
Jesuit Yves de la Briere and writer Georges Duhamel (see Cattani, 2017, pp. 675, 677). 5.3. A Narrowing Horizon of Expectation As long as there is a democratic regime in Germany,
Coudenhove-Kalergi deems it is not too late for reconcili-
ation. Nevertheless, he is pessimistic about the time that
is left (Coudenhove-Kalergi, 1923, p. 131). His book aims
to underscore the urgent need of European unity, before
the horizon of expectation will narrow again. The United
States of Europe is “the only salvation from the chaos of
the present—before the collapse of the future” (p. 39). To pave the way for reconciliation and understanding
would require a magnanimous, generous and redemp-
tive act (grossmütiger Akt) of France to “teach Germany
to believe in the French will to reconciliation to strike
a deadly blow to the politics of revenge” (Coudenhove-
Kalergi, 1923, p. 131). In particular, Coudenhove-Kalergi
thinks of “a voluntary renunciation of the occupation of
the Rhine” in exchange for an “inter-European guarantee
treaty” (1923). This shows that, in Coudenhove-Kalergi’s
understanding, pan-Europe provides the necessary con-
ditions for a politics of reconciliation. 5.2. Pan-Europe: Europe of the Future—Reconciliation &
Understanding Against this grim picture, Coudenhove-Kalergi proposes
his alternative, almost eschatological, horizon of salva- Politics and Governance, 2018, Volume 6, Issue 4, Pages 135–143 139 tion, or redemption (Rettung), liberation and solidar-
ity: European economic integration, based on a political
union (1923, pp. IX, 27, 70). The pan-European horizon
that Coudenhove-Kalergi aims to provide would be based
on a French politics of reconciliation (Versöhnung) and
a German politics of understanding (Verständigung) (pp. 124–128). This almost eschatological and salvific hori-
zon unites the “fallen men” of Europe with their once
so greatly ordained destiny; thereby channeling a typi-
cal sense of Christian teleology into the political realm. To step towards this horizon, this Christian notion of re-
demption and reconciliation also entails a break with
the past; redemption does not arrive automatically—the
way needs to be paved with sacrifices and personal en-
gagement (1923, p. 71). would bring about one’s own ruin, would suffice in first
instance (1923, p. 124). In his opposition of anti-European and Pan-European
Coudenhove-Kalergi is careful not to position national
identity against a pan-European identity. Rather, he
refers to Pan-Europe as Übervaterland (1923, p. 156). In
later writings he further expands on this relationship be-
tween European and national identity, arguing that “just
as love for the mother does not detract love for the fa-
ther, neither does love for the European Mutterland re-
duce love for the national Vaterländern” (Coudenhove-
Kalergi, 1931, p. 17). So, national and European identity
are no zero-sum relationship (perhaps: “are not mutually
exclusive”). This image of the family has a strong emo-
tional connotation, referring to a given relationship—
whether you like it or not. Against the background of the
suffering caused by WWI, the maternal reference to Eu-
rope underlines its emotional and pacifist connotation
(Cattani, 2017, pp. 677–678)8. The notion of sacrifice was a key emotive used as
part of the war culture rhetoric (Horne, 2004) that
Coudenhove-Kalergi is so strongly opposing. Rather than
referring to sacrifice as a way to claim moral compen-
sation and renewing antagonism, Coudenhove-Kalergi
uses this vocabulary in combination with the notion
of reconciliation. 6. Emotionalizing Effects, Reception & Response Forward-looking Europe
salvaon, liberaon,
reconciliaon,
understanding, solidarity Forward-looking Europe
salvaon, liberaon,
reconciliaon,
understanding, solidarity Backward-looking
Europe salvaon, liberaon,
reconciliaon,
understanding, solidarity Figure 1. Emotional discourse Pan-Europa. emotives to mobilize his audience, it was not success-
ful in preventing another vicious cycle of French-German
politics of hate. As such, this article highlights the con-
straining force of emotives of enmity on what can be
imagined as possible horizons. to change the course of history. As Coudenhove-Kalergi
(1943) himself points out in his memoirs, his movement
was supported by many individual members of govern-
ments, but at a more official level, governments ignored
it. Opposition did not only come from nationalist groups,
but also from pacifists accusing Coudenhove-Kalergi
of advocating a Machiavellian pacifism (Prettenthaler-
Ziegerhofer, 2013, p. 163). Coudenhove-Kalergi (1943)
recalls that he had not expected Hitler to take over
Germany: “When I came back to Germany in 1924, the
Hitler chapter seemed definitely closed”. Nevertheless,
his fear of another cycle of rage and destruction politics
came true. to change the course of history. As Coudenhove-Kalergi
(1943) himself points out in his memoirs, his movement
was supported by many individual members of govern-
ments, but at a more official level, governments ignored
it. Opposition did not only come from nationalist groups,
but also from pacifists accusing Coudenhove-Kalergi
of advocating a Machiavellian pacifism (Prettenthaler-
Ziegerhofer, 2013, p. 163). Coudenhove-Kalergi (1943)
recalls that he had not expected Hitler to take over
Germany: “When I came back to Germany in 1924, the
Hitler chapter seemed definitely closed”. Nevertheless,
his fear of another cycle of rage and destruction politics
came true. This emotion discourse analysis of Pan-Europa has
been a limited exercise. There are, at least, three ways
to further expand on this analysis. First, compare Pan-
Europa with Coudenhove-Kalergi’s emotional vocabulary
after the war. This would allow to reflect upon the rela-
tionship between emotives and crisis. Second, compare
Pan-Europa with the emotional vocabulary of contem-
poraries (e.g., Briand’s Federal European Union, 1930;
Rohan’s Europa, 1924). It would allow one to assess to
what extent and how Coudenhove-Kalergi’s notions of
reconciliation and solidarity resonated with contempo-
rary political elites, and to what extent religiously in-
spired idiom still could find a home in European politics. As Frevert (2015) shows, the Nazi politics of emotions
drew also on religious semantics to mobilize its audience. 6. Emotionalizing Effects, Reception & Response Third, methodologically it would be of interest to inves-
tigate in what ways an analysis of the images (i.e., book
covers, symbols) confirm the emotion discourse analysis. 6. Emotionalizing Effects, Reception & Response Central to Pan-Europe, and Coudenhove-Kalergi’s writ-
ings in general, is its rather dichotomous character. Coudenhove-Kalergi integrates a sense of time with
particular emotives and emotional beliefs. The anti-
European option is based on backward-looking emo-
tives of enmity, whereas the pan-European order is re-
ferred to in terms of forward-looking emotives of amity
(Figure 1). The notion of reconciliation is of course closely con-
nected to that of sacrifice (Opfer): Those who love Europe must not shy away from any
sacrifice in order to save their continent from this mor-
tal danger. (Coudenhove-Kalergi, 1923, p. 97, author’s
own translation) This quote also underlines the point that while
Coudenhove-Kalergi speaks about the relationship be-
tween France and Germany at a state-level, he also calls
on every individual citizen to assume its responsibility. Neutrality in this question of “life and death” is not pos-
sible, running away from a decision is high treason (1923,
p. 102). Moreover, it recognizes the feelings of enmity. Coudenhove-Kalergi did recognize the dominance
of emotives of enmity among his contemporaries and
aimed to expand their horizon of expectation to con-
ceive of a different European order. At his Pan-European
Congresses he aimed to practise these notions of
reconciliation—a highly Christian infused and morally
laden emotive—and solidarity—a more social and sec-
ularized one. When the German government discussed
whether to officially participate in the 2nd Pan-European
Congress in Berlin in 1930, Minister of the Interior Wirth,
argued in favour with reference to this notion of recon-
ciliation (Prettenthaler-Ziegerhofer, 2001). To
break
the
resistance
against
Pan-Europa,
Coudenhove-Kalergi looks for an awakening (Erwachen)
of a pan-European feeling of solidarity (1923, p. 109). This awakening suggests that solidarity is already there,
it only has to be activated. Coudenhove-Kalergi distin-
guishes between two types of solidarity—a solidarity of
love and a solidarity of reason. He primarily refers to
the latter, i.e., the awareness that the ruin of the other Although
Coudenhove-Kalergi
strongly
connects
emotion to mobilize forces for his Pan-European or-
der, his Pan-European movement has not proved able Politics and Governance, 2018, Volume 6, Issue 4, Pages 135–143 140 Backward-looking
Europe
hate, rage, revenge,
destrucon, naonalism
Forward-looking Europe
salvaon, liberaon,
reconciliaon,
understanding, solidarity
Figure 1. Emotional discourse Pan-Europa. Backward-looking
Europe
hate, rage, revenge,
destrucon, naonalism
Figure 1. Emotional discourse Pan-Europa. 7. Conclusion While research on the interwar plans for European inte-
gration is well developed, much less is known about the
emotive vocabulary that is involved in those “blueprints
of Europe”. To this end, this article analyzed Coudenhove-
Kalergi’s canonical text Pan-Europa. Central to his experience is WWI and the hate be-
tween Germany and France. This hate is combined with
a strong inclination to look back. As such it leads to a vi-
cious circle of destruction and revenge. These negative
emotives associated with nationalism as emotional be-
lief feed into Coudenhove-Kalergi’s plan for an alterna-
tive European political order which is centered on the
emotional and spiritual belief of reconciliation, sacrifice
and solidarity—invoking a moralistic appeal and call to
action. Coudenhove-Kalergi uses fear of the past to urge
his audience to expand their horizon of expectation, and
to mobilize their inner redemptive drive. He aims to ex-
pand this horizon of expectation with an almost eschato-
logical plan for European unity, in which these negative
emotions would turn into feelings of solidarity. While he
acknowledges that a solidarity of love may be a bit too
much, he calls for a solidarity of reason—and for great
acts of sacrifice to meet these high-pitched stakes. Acknowledgments This work has been supported by the Netherlands Orga-
nization for Scientific Research (NWO) under grant #360-
52-190. I would like to thank the members of the re-
search group Blueprints of Hope: Prof. Beatrice de Graaf
(PI), Prof. Peter-Ben Smit, Prof. Mathieu Segers, Clemens
van den Berg and Jorrit Steehouder. Moreover, the arti-
cle benefitted from comments from Prof. Eckhart Conze
(May 2017) and comments received at EUSA Miami
(May 2017). The author declares no conflict of interests. The author declares no conflict of interests. Using Koselleck’s (2005) meta-historical categories of
space of experience and horizon of expectation allows
one to contextualize this emotive vocabulary. We have
seen how Coudenhove-Kalergi aimed to connect the
emotive of sacrifice to a politics of reconciliation, rather
than destruction and resentment. While Coudenhove-
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in Cambridge Review of International Affairs, Scandinavian Political Studies and Contemporary Politics. 143
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EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE: APLICABILIDADE DAS NOVAS TECNOLOGIAS
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v.2, n.7, 2021 v.2, n.7, 2021 EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE: APLICABILIDADE DAS NOVAS
TECNOLOGIAS
EDUCATION 4.0 AND HOW NURSING IS INSERTED: APPLICABILITY OF NEW TECHNOLOGIES Leticia Falcão Nogueira1, Emanuel Pereira dos Santos2, Claudiane Blanco Andrade dos Santos3, Vera Lúcia
Freitas4 e27589
https://doi.org/10.47820/recima21.v2i7.589 KEYWORDS: Education 4.0. Industrial Revolution. Technological innovations. Nursing. History of
nursing. Applications. Continuing education in nursing. RESUMO A presente resenha crítico-descritiva traz a análise de um paralelo entre a Educação 4.0 e a
Revolução Industrial e como elas impactam no cuidar, relacionar e aprender da Enfermagem, mais
ainda em se tratando de um contexto da COVID-19, onde tais mudanças ocorreram de forma tão
acelerada. Tem como cerne, trazer à luz a análise da evolução industrial e como ela contribui com as
mudanças na forma de aprender e ensinar, bem como, analisar as linguagens necessárias na
revolução 4.0 para lecionar conteúdos de maneira predominantemente on-line. A tabela apresentada
no estudo facilita a visualização dos novos usuários das tecnologias que são tão pertinentes para o
aprendizado e para o trabalho, desta forma ajudamos as pessoas a inovarem cada vez mais,
respeitando as características únicas em relação ao ritmo de aprendizagem do indivíduo. PALAVRAS CHAVES: Educação 4.0. Revolução Industrial. Inovações tecnológicas. Enfermagem. História da enfermagem. Aplicativos. Educação continuada na Enfermagem. 1 Graduanda de Enfermagem da Universidade Federal do Estado do Rio de Janeiro (UNIRIO) RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia
Enfermeiro Mestre pela Universidade Federal do Estado do Rio de Janeiro (UNIRIO)
3 Advogada graduada em Direito e Pós-graduada em Direito Processual Civil pela Universidade Candido Mendes
(UCAM)
4 Docente Doutora na Universidade Federal do Estado do Rio de Janeiro (UNIRIO) g
(
)
2 Enfermeiro Mestre pela Universidade Federal do Estado do Rio de Janeiro (UNIRIO) RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia Enfermeiro Mestre pela Universidade Federal do Estado do Rio de Janeiro (UNIRIO)
3 Advogada graduada em Direito e Pós-graduada em Direito Processual Civil pela Universidade Candido Mendes
(UCAM)
4 Docente Doutora na Universidade Federal do Estado do Rio de Janeiro (UNIRIO) (UCAM)
4 Docente Doutora na Universidade Federal do Estado do Rio de Janeiro (UNIRIO) p
(
)
da graduada em Direito e Pós-graduada em Direito Processual Civil pela Universidade Candido Mendes 1 Graduanda de Enfermagem da Universidade Federal do Estado do Rio de Janeiro (UNIRIO)
2 E f
i
M
t
l
U i
id d
F d
l d
E t d
d
Ri
d
J
i
(UNIRIO) p
(
)
3 Advogada graduada em Direito e Pós-graduada em Direito Processual Civil pela Universidade Candido
(UCAM) 1 Graduanda de Enfermagem da Universidade Federal do Estado do Rio de Janeiro (UNIRIO)
2 Enfermeiro Mestre pela Universidade Federal do Estado do Rio de Janeiro (UNIRIO)
3 Advogada graduada em Direito e Pós-graduada em Direito Processual Civil pela Universidade Candido Mendes
(UCAM)
4 Docente Doutora na Universidade Federal do Estado do Rio de Janeiro (UNIRIO) ABSTRACT This descriptive critical review deals with a parallel between the Education 4.0 Industry Revolution and
how they impact the care, relationship and learning of Nursing, even more in the context of Covid-19
where such changes occurred so fast. Its core is to bring to light the analysis of industrial evolution
and how it contributes to changes in the way of learning and teaching, as well as analyzing the
languages needed in the 4.0 revolution to teach content predominantly online. The table presented in
the study facilitates the visualization of new users of technologies that are so relevant for learning and
for work, in this way we help people to innovate more and more, respecting the unique characteristics
in relation to the individual's learning pace. KEYWORDS: Education 4.0. Industrial Revolution. Technological innovations. Nursing. History of
nursing. Applications. Continuing education in nursing. RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia v.2, n.7, 2021 RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas INTRODUÇÃO As escolas necessitam preparar estudantes para trabalhos que ainda não foram criados, para
tecnologias que ainda não foram inventadas, para resolver problemas que ainda não foram previstos
(OECD, 2018). A Educação 4.0 pode ser vista como uma nova interpretação dos conceitos como
aprendizagem, aluno, professor e escola, alinhados com as necessidades da Indústria 4.0
(HIMMETOGLU et al.,2020). O uso das tecnologias da quarta revolução industrial se torna
imprescindível para a contínua evolução nas formas de relacionar, cuidar e aprender. A enfermagem,
como sempre se encontra no centro do Ato de cuidar, logo, inovar desde a graduação se torna
indispensável. Com o COVID-19, houve uma aceleração em tais modificações que estavam
acontecendo de forma mais lenta desde a globalização. As escolas e faculdades tiveram que aderir a
ferramentas para facilitar a aula on-line, os alunos tiveram que aprender novas tecnologias para
apresentações de trabalho e para uma melhor interação com os docentes, tal panela de pressão foi o
cenário perfeito para radicalizar a dinâmica do aprender, estudar e consequentemente trabalhar
(CARMO et al., 2020). Diante desse cenário se faz necessária a adequação dos profissionais para
utilizarem essas plataformas como instrumento para o ensino, desenvolvendo competências e
usando de forma racional os recursos disponíveis. Dado o avanço da tecnologia, com a grande demanda por uso de aplicativos na atualidade, e como
reflexo dessa iteratividade, também no ambiente acadêmico, logo percebe-se a extrema importância
de reunir todos esses aplicativos utilizados, elencados em um quadro de modo a facilitar a
visualização e a manejo pelos usuários, quer sejam eles graduandos, pós-graduandos, docentes e
técnicos administrativos. Esse quadro possibilitará a futuros usuários dessas novas ferramentas de
aprendizado, que são os aplicativos, a escolha de um melhor método de interação, conforme haja a
necessidade de sua utilização no universo acadêmico. Dependendo do cenário apresentado na Universidade, o programa de computador ou aplicativo de
celular (que pode apresentar a mesma ferramenta em dispositivos diferentes), pode necessitar de
uma especificidade diferente para auxiliar no processo ensino/aprendizagem. O quadro de
apresentação das ferramentas é uma excelente opção para mostrar aos usuários as escolhas
disponíveis a serem usadas que mais se adequem à sua necessidade de uso, uma vez que, ao se
propor uma inovação tecnológica, um importante alavanca de mudanças é acionada (PANIAGUA,
2018). Segundo Pinto et al. (2021), Educação 4.0 é: Segundo Pinto et al. RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia INTRODUÇÃO (2021), Educação 4.0 é: “O modelo entende que o uso irrestrito dos recursos tecnológicos visando a
interação irá proporcionar o desenvolvimento da coletividade a partir das
contribuições mútuas e coletivas, estimulando a criatividade e as competências
sócioemocionais”. 2 2 2 v.2, n.7, 2021 RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas Segundo Pinto et al.(2021): Segundo Pinto et al.(2021): “educação 4.0 vem se tornando uma necessidade premente nas diversas áreas do
saber. As profissões perpassaram a necessidade básica do conhecimento,
carecendo uma maior gama de informações, ações, comportamentos e
relacionamentos individuais e coletivos para o correto desempenho de suas
atividades”. RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas da eletricidade e da energia química, os grandes motores. Tais elementos possibilitaram um aumento
substancial na produção de insumos, o que precisou de um estímulo ao consumo considerável para
manter a cadeia de produção por linhas de montagem. No cenário educacional, a Segunda
Revolução industrial ensejou na vasta utilização dos quadros negros e de laboratórios químicos
escolares (HARKINS, 2008; SCHWAB e DAVIS, 2018). da eletricidade e da energia química, os grandes motores. Tais elementos possibilitaram um aumento
substancial na produção de insumos, o que precisou de um estímulo ao consumo considerável para
manter a cadeia de produção por linhas de montagem. No cenário educacional, a Segunda
Revolução industrial ensejou na vasta utilização dos quadros negros e de laboratórios químicos
escolares (HARKINS, 2008; SCHWAB e DAVIS, 2018). Os autores referenciados, ao falarem da Segunda Revolução Industrial, se referem à qualidade da
energia empregada na produção desses produtos. Isso nos mostra que, quão melhor a qualidade da
ferramenta a ser utilizada, maior será o produto. Em se tratando da Educação 4.0, quanto melhor o produto, maior a parcela populacional atingida e
melhor o processo de ensino e aprendizagem observado em questão. Por outro lado, na Terceira Revolução Industrial, Harkins (2008) a categoriza como aquela
responsável pela globalização mundial em uma escala astronômica, quando comparada com as
Grandes Navegações do século XV. Com o Surgimento da Rede Mundial (Internet), que possibilitou maior troca de informação e
comunicação em toda a parte do mundo, encurtando distâncias, essa nova realidade inaugurou
mudanças reais nas relações do trabalho e no modo de produção quando comparada à Segunda
Revolução. Tecnologias eletrônicas de ponta, novos métodos de conexão, estabilidade de conexão em maiores
distâncias, usos de satélites, comunicação em escala mundial, o aprimoramento da linguagem
internacional adotando o inglês como língua universal, dentre tantos outros avanços, transformaram
significativamente o cenário mundial através do avanço tecnológico, tornando uma ambiência
propícia para receber a Revolução 4.0 atualmente vigente. Várias linguagens foram necessárias até a atual Revolução 4.0. O mundo necessitou se conectar por
cabos e sinais de antenas e satélites. REVISÃO TEÓRICA Historicamente o desenvolvimento das chamadas tecnologias digitais de informação e comunicação
(TDIC's) trouxe à Educação desdobramentos no que se referem à formação docente e acadêmica. Assim, tal desenvolvimento somente foi possível devido ao capital investido em pesquisa e em
conhecimento. Com o investimento do capital em pesquisa, muitos avanços puderam ser observados. Esta
perspectiva relaciona-se à uma evolução histórica da indústria tecnológica. Segundo Harkins, (2008) a economia - neste trabalho incluindo todas as formas de tecnologia que
foram incorporadas à produção - pode ser classificada segundo seu período histórico e as formas de
energia utilizadas no processo de produção. Essa nova forma de interagir, através do uso de aplicativos, possibilita a diminuição do tempo
consumido pelo usuário, consequentemente, permitindo-lhe melhor qualidade de tempo. Toda nova
tecnologia tem a proposta de melhoria de algum aspecto que esteja dificultado em sua sociedade ou
na sociedade de uma forma mais globalizada, onde se observa a necessidade em diversas culturas. O Grande marco tecnológico da primeira Revolução Industrial foi a forma de fabricação dos produtos,
que antes eram feitos por artesãos em suas residências, passando a serem feitos por maquinários
movidos a energia a vapor, possibilitando assim uma produção em escala maior. Na educação, o
desafio foi transformar esses artesãos em operários, para comporem a força de trabalho nas fábricas,
muitas vezes colocando-os em salas de aula com quadros negros para aprenderem o ofício
(SCHWAB, 2016; SCHWAB e DAVIS, 2018). Conforme análise trazida pelos autores referenciados acima, a Revolução Industrial chegou para
modificar a forma de produção e seus processos de construção, com o intuito de maximizar a
produção, possibilitando assim, a oferta de produtos e serviços em escalas maiores. Trazendo essa
mesma linha de raciocínio à educação, torna-se uma ferramenta que viabiliza o acesso em escalas
maiores a uma maior parcela da população. Na Segunda Revolução Industrial, tem-se não somente como elemento principal o produto. Nesse
cenário, tem-se a atenção voltada ao tipo de energia utilizada. Nesta nova fábrica predominam o uso RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia 3 3 3 v.2, n.7, 2021 RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 Para que vários pontos de nosso planeta com culturas e
línguas diferentes pudessem ser conectados, uma língua teve que sobrepujar sobre as outras, no
intuito do entendimento comum e, atualmente, o uso de cabos, por exemplo, se vê desnecessário,
tamanha a velocidade no aperfeiçoamento das tecnologias ao longo do tempo. RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia METODOLOGIA O estudo em tela, trata-se de uma resenha crítica descritiva. Segundo (BELMONT, 2004) “Resenhar
é relatar as partes que constituem um objeto, observando seus aspectos mais relevantes”. Nesse
estudo serão apresentadas as principais ferramentas que podem ser utilizadas no meio acadêmico. v.2, n.7, 2021 RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas O quadro apresentará, de forma descritiva, as possibilidades a serem utilizadas no momento da
formalização do ensino/aprendizado através do uso das ferramentas tecnológicas vigentes até o
momento. A estratégia de busca dos artigos foi realizada com as seguintes palavras-chave: (Educação 4.0;
Revolução industrial; Inovações tecnológicas; Enfermagem; História da enfermagem; Aplicativos;
Educação continuada na enfermagem) em quatro bases de dados, sendo elas: Medline (0), Lilacs (1),
Google Acadêmico (6), Scielo (3). Os seguintes artigos serão tabulados e discutidos no próximo item. A busca e leitura dos referidos materiais foi realizada entre junho e julho de 2021, onde a escrita e
análise foram feitas minuciosamente. A análise dos artigos ajudará os leitores a selecionarem os
melhores aplicativos/programas a serem utilizados na relação ensino/aprendizagem. Não houve necessidade da apreciação do CEP (Comitê de Ética em Pesquisa), pois a resolução Nº
466 de 12 de Dezembro de 2012 não prevê que seja necessária apreciação no caso de resenhas de
materiais já publicados, uma vez que não há nada notificado sobre o assunto na resolução citada. RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia DISCUSSÃO/APRESENTAÇÃO DOS DADOS Estão sendo apresentadas no quadro abaixo, as ferramentas com as possibilidades de utilização
para auxiliar na escolha da ferramenta mais adequada para a atividade pedagógica a ser utilizada. É
importante ressaltar, que para cada perfil de aluno e cada perfil de proposta de aprendizado, uma ou
mais ferramentas podem ser utilizadas entre o professor e o aluno. As ferramentas auxiliarão no
aprendizado. Cabe ao educador escolher quais aplicativos de celular/programas de computador serão mais
proveitosos em suas aulas, palestras ou cursos oferecidos durante o período letivo ou até mesmo
fora deles. O quadro a seguir, apresenta o tipo de função que pode ser explorada pelo educador, o
seu conceito de forma resumida para um bom entendimento e os exemplos mais utilizados nessa
época. Dada a crise sanitária mundial, ocasionada pela pandemia de COVID-19, muitos alunos e
professores necessitaram realizar suas atividades de casa, o que ensejou no desenvolvimento e na
maior utilização da modalidade de ensino de natureza exclusivamente on-line. RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia 5 5 v.2, n.7, 2021 RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218
EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas
Quadro 1 – Ferramentas para Educação on-line. FUNÇÃO
CONCEITO
EXEMPLO
Criação de conteúdo
de vídeo interativo
É possível criar conteúdos e vídeos
interativos
de
forma
simplificada,
gerando uma apresentação interativa e
dinâmica
do
conteúdo
a
ser
apresentado. YouTube Vídeo Editor;
Criação
de
Infográficos
e
cartazes. A utilização de Infográficos e cartazes
auxilia no entendimento mais detalhado
de um conteúdo mais complexo, pois é
a junção de um desenho ou imagem
com o auxílio de um texto utilizado para
explicar ou informar sobre o assunto que
não seria tão bem compreendido. São
ótimas ferramentas para criação em sala
de aula ou Laboratórios de Informática. Piktochart;
Infogram;
Easel;
Canva;
Criação
de
Blogs,
wikis para espaços
participativos
para
estudantes. O Blog é uma ferramenta muito útil para
criar
um
espaço
de
ensino
e
aprendizagem
onde
alunos
e
professores têm acesso mutuamente e
poderão contribuir mutuamente para a
evolução acadêmica ou até mesmo para
ministrar
cursos
virtuais
de
aprimoramento de técnicas do ensino
prático. Wordpress;
Edublogs;
Wikia;
Google Classroom;
Criação
de
apresentações. RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia DISCUSSÃO/APRESENTAÇÃO DOS DADOS Em aulas expositivas e dialogadas, a
forma de transmitir o conteúdo é muito
importante para prender a atenção. Prezi;
Canva;
Fppt;
Flips-Nack; EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas 6 v.2, n.7, 2021 RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas Salas de aula virtuais, como o próprio
nome diz, vêm para simular a vivência
da sala de aula à distância. Tentando
manter a mesma pessoalidade da aula
presencial sem deixar de ser atual e
inovadora. Criação de salas de
aula virtuais. Rede escola digital; Criação de salas de
aula virtuais. Fonte: Nogueira, L. F.; Santos, E. P.; Santos, C. B. A.; Freitas, V. L. Educação 4.0 e como a
Enfermagem se insere: Aplicabilidade das novas tecnologias. Monografia de Graduação da
EEAP/UNIRIO 2021. O aprendizado de forma on-line possibilita que pessoas que não podem se encontrar fisicamente por
diversos motivos (pandemia, barreiras geográficas e outros limitadores presenciais), possam
continuar aprendendo e produzindo conhecimento de forma a auxiliar na inclusão nesses casos,
onde anteriormente seria inviável a participação desse processo ensino/aprendizado pela ausência
dessa opção tecnológica. Segundo Sonego e Behar (2019), “acredita-se que existe um leque de possibilidades para a
realização de atividades com os smartphones e tablets, potencializando o desenvolvimento da m-
learning". Ao longo das últimas duas décadas, nossa sociedade teve um grande avanço tecnológico,
culminando não somente com a internet, como também a criação e aperfeiçoamento de novos
dispositivos que a recebem, aumentando as possibilidades de acesso. Atualmente, pode-se realizar estudos através de diversos dispositivos que, no início dos anos 2000,
não se poderia prever que em poucos anos estariam disponíveis para uso por grande parte da
população. Segundo Sonego e Behar (2019), “Acerca desse termo, existem muitas discussões em
nível mundial, que abordam que, dentre os dispositivos, devem ser considerados os equipamentos do
tipo, smartphones, tablets, Ipads, notebooks e televisão do tipo smart tv”. Ainda que a necessidade de adaptação das instituições de ensino para continuar ministrando seus
conteúdos letivos tenha se dado em decorrência da pandemia, essa nova tendência de estudo à
distância já vinha sendo implementada aos poucos em nossa sociedade. Deve-se destacar que o aprendizado apresentará diversas transformações, conforme a sociedade
mostre mais necessidade de investimento em mais tecnologia, ou mesmo pela necessidade de
adequação aos novos pensamentos que surgirão ao longo dos anos. 7 v.2, n.7, 2021 RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas Segundo Sonego e Behar (2019), “destaca-se que a todo o momento surgem novas tendências em
tecnologias. Com isso, são verificadas novas demandas dos alunos, para que o uso dos dispositivos
seja efetivado através de atividades escolares”. O docente já possui um papel muito importante no aprendizado dos seus alunos mesmo antes da
globalização, que popularizou a Internet e sua utilização em larga escala. Com essa nova de
adaptação às plataformas digitais pelas instituições de ensino, o docente, a partir de então, deverá
estar sempre atento às possíveis características decorrentes de diferenças socioeconômicos que
seus alunos apresentem, de modo a possibilitar que todos sejam efetivamente incluídos e possam
participar dessa nova realidade que acortinou-se para a educação mundial. É importante trazer à discussão, que ainda que se esteja diante de um cenário necessariamente
virtual entre alunos e professores, não se pode fechar os olhos para as diferentes realidades
ocasionadas por barreiras socioeconômicas, que ao longo da pandemia, têm inviabilizado o acesso
ao ensino de qualidade através das plataformas digitais, por grande parte dos estudantes brasileiros,
abarcando tanto os níveis de pós-graduação, graduação, como também alunos dos níveis médio e
fundamental. O acesso aos conteúdos digitais não é uniforme, tampouco linear para todos os alunos e professores,
visto que diversas sãos as barreiras, que vão desde à impossibilidade de fazerem uso de um
dispositivo capaz de suportar o acesso às plataformas digitais, como também a limitação na conexão
com a Internet, para acessar os conteúdos educacionais. RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas “Neste sentido, entendemos que as implicações da inclusão e da exclusão digital na
formação de educadores possuem desdobramentos impactantes nas práticas
pedagógicas,
principalmente
quando
consideramos
que
nem
todos
os
educadores/as participaram de processos de inclusão digital na perspectiva aqui
apontada, a partir da realidade concreta apresentada nas pesquisas”. A discussão sobre tecnologias para substituir a necessidade de o aluno estar em sala de aula ainda é
recente e muitas instituições não se utilizavam, de forma expressiva, dessa tecnologia. A discussão sobre tecnologias para substituir a necessidade de o aluno estar em sala de aula ainda é
recente e muitas instituições não se utilizavam, de forma expressiva, dessa tecnologia. Os professores necessitam de investimento necessários para que possam suportar esse novo
cenário, através de equipamentos e capacitação de modo que seja ofertada uma atualização, uma
vez que limitar-se a apresentar as novas ferramentas para pessoas que não têm qualquer intimidade
com dispositivos eletrônicos, torna muito difícil a sua incorporação de forma eficaz no dia a dia. Em
razão disso, faz-se necessária a instrumentalização através da capacitação, de modo a garantir o
melhor uso possível das ferramentas tecnológicas, nesse processo de inovação das instituições de
ensino. Segundo Sonego (2019), “Fornecer apoio e formação a professores por meio de tecnologias móveis:
para realizar a incorporação dos dispositivos móveis nas ações docentes, se faz necessário que o
professores tenham perpassado por formação pedagógica inicial e continuada específica para o uso
destes dispositivos”. Os dispositivos tecnológicos apresentam uma gama de possibilidades para tanto os alunos, quanto
os professores e os funcionários da educação, e até mesmo para profissionais da área da saúde, por
meio da realização de buscas acadêmicas e de suporte nos estudos que viabilizam a continuidade do
acesso ao conhecimento e a construção do saber. Segundo Sonego e Behar (2019), “percebe-se que um aplicativo pode apresentar inovações nas
práticas docentes, com possibilidades de pesquisa, escrita, leitura, construção de conteúdo, além de
seu compartilhamento entre os envolvidos nesse processo”. Segundo (SOUZA & GUIMARÃES, 2020) Segundo (SOUZA & GUIMARÃES, 2020) “Essa imensa desigualdade no acesso à internet adquire novos contornos diante da
pandemia de Covid-19 pois, em razão do isolamento social recomendado pelas
autoridades sanitárias, os sistemas de ensino passaram a adotar a educação a
distância, que, da forma como foi implantada, tem se revelado como mais um
indicativo da desigualdade social que sempre caracterizou a educação brasileira”. Também merece atenção a realidade dos docentes, que, em sua maioria, tiveram uma formação
onde esses dispositivos não eram utilizados. Segundo Albino (2019): “O docente na educação 4.0 tem um papel mais amplo no processo de ensino
aprendizagem pois deve variar suas metodologias para conseguir atingir o máximo
entendimento por parte dos alunos sabendo de suas diferenças sociais e culturais,
desta forma o docente deve buscar novos conhecimento e ferramentas que auxilie
nesse processo, pois em alguns momentos será o transmissor do conhecimento, em
outro o mediador, e até mesmo o orientador de pesquisas direcionadas utilizando
ferramentas por ele indicada”. Segundo Marcon (2020): RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia 8 8 8 v.2, n.7, 2021 RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 Após o fim dessa pandemia de COVID-19, o mundo retornará às salas de aula, porém, ocorrerá a
possibilidade do aprendizado híbrido, onde as salas de aula poderão ser físicas e, em alguns
momentos, virtuais, conforme a necessidade de cada grupo acadêmico. Segundo Albino (2019), “O aprendizado híbrido acontece tanto em uma sala de aula (ou outro espaço
físico) quanto on-line". A modalidade forma híbrida de estudo, se dará com o aluno podendo realizar parte do seu estudo
estando fisicamente dentro das instalações da instituição de ensino, e o professor, dentro de casa, ou
mesmo em uma sala de aula padrão invertida, onde se poderá ser feita de casa, com conteúdos
gravados para serem acessados no momento que melhor couber aluno, permitindo-lhe administrar o
seu próprio tempo. RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia 9 9 9 v.2, n.7, 2021 RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas Segundo Albino (2019), “A sala de aula padrão invertida: os alunos recebem a lição em casa e
assistem a palestras em vídeo e devem ler qualquer material relevante para a aula do dia seguinte”. CONCLUSÃO A Educação 4.0 com suas propostas e inovações, garantem aos professores e alunos maior contato
no processo da construção do conhecimento, respeitando as características únicas do indivíduo em
relação ao ritmo e características de aprendizado, como também o caráter socioeconômico em que
cada indivíduo se encontra. Os aplicativos utilizados para facilitar a boa adesão do conhecimento do conteúdo trabalho são
vários, se fazendo necessário o uso de criatividade, investimento e capacitação técnico dos
profissionais, uma vez que muitas são as ferramentas que vêm surgindo e que estão à disposição
dos alunos e professores, de modo a viabilizar o acesso à educação à distância. Com esse compilado de aplicativos/programas espera-se poder servir de auxílio a acadêmicos,
docentes e profissionais a se atualizarem a fim de gerar um ambiente de aprendizado mais dinâmico
e inclusivo. Da mesma forma que nas Revoluções Industriais apresentaram mudanças sociais geradas através
dos novos modelos de produções e das novas formas de ensinar e de reter conhecimento
culminando em mudanças significativas na sociedade, tivemos um desdobramento dessa atualização
nas práticas de ensino e aprendizagem no contexto gerado pela COVID-19 quando escolas,
universidade e ambientes de trabalho precisaram ficar necessariamente remotos, exigindo de
professores e gestores manejo na administração de contatos com alunos e funcionários de modo a
continuarem produzindo. Tal cenário desencadeou uma ambiência favorável para a criação e utilização de aplicativos e
programas com propósitos facilitadores de ministração de conteúdos assim como também deflagrou
as diferenças socioeconômicas como desafios para o acesso às plataformas digitais por alunos e
professores. A nova oportunidade criada pela pandemia ofereceu a possibilidade de desenvolver de forma maciça
a educação a distância, permitindo que possa alcançar a todos os alunos e professores, mostrando a
necessidade de buscar desenvolver cada vez mais, ferramentas que possibilitem o acesso à
educação também, pelo meio virtual. Faz-se necessário portanto, criar uma nova consciência de adaptação às novas realidades que se
apresentem, bem como desenvolver meios de manutenção de informação e formação, ainda que de
forma remota. RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia 10 10 v.2, n.7, 2021 RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas Uma educação que promove maior alcance de estudantes é, de forma direta, mais inclusiva, pois as
distâncias entre as residências e as instituições de ensino são encurtadas, possibilitando os que têm
mais dificuldades de acesso, principalmente pela distância, poderem dispor da oportunidade de
continuar os seus estudos com a diminuição da evasão escolar por tempo de transporte ou a
impossibilidade de locomoção, que no momento se deflagrou com a pandemia mundial. Se houver união de esforços, investimento, redução de custos e oferta de dispositivos de acesso à
internet, conexões viáveis disponíveis de modo equânime, tanto a alunos quanto a professores, é
possível se oferecer uma educação de qualidade, que chegue a mais pessoas. REFERÊNCIAS ALBINO, R. As Principais Metodologias e Ferramentas na Educação 4.0. Araçatuba, SP: s.n.,
2019. ALBINO, R. As Principais Metodologias e Ferramentas na Educação 4.0. Araçatuba, SP: s.n.,
2019. ARRUDA, E. P. Educação Remota Emergencial: elementos para políticas públicas na educação
brasileira em tempos de Covid-19. Em Rede - Revista de Educação a Distância, v. 7, n. 1, 2020. CARMO, J. R.; PACIULLI, S. O. D.; NASCIMENTO, D. L. O impacto do uso de Tecnologias de
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em um contexto de pandemia. Research, Society and Development, [S. l.], v. 9, n. 10, p. e5199108940, 2020. COSTIN, C. Educar para um futuro mais sustentável e inclusivo. Estudos Avançados [online], v. 34,
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em: 29 jul 2021. HARKINS, A. M. Leap frog principles and practices: core components of education 3.0 and 4.0. Futures research quarterly draft VIII. 2008. HIMMETOGLU, B.; AYDUG, D.; BAYRAK, C. Education 4.0: Defining the teacher, the student, and
the school manager aspects of the revolution. Turkish Online Journal of Distance Education, v. 21,
(Special Issue-IODL), p. 12-28, 2020. INTERNATIONAL LABOUR ORGANIZATION - Global Commission on the Future of Work-2019. Work for a Brighter Future. Genève: International Labour Office, 2019. KAGERMANN, H.; WAHLSTER, W.; HELBI, J. Recommendations for implementing the strategic
initiative INDUSTRIE 4.0. Frankfurt: National Academy of Science and Engineering/Federal Ministry
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2018 RECIMA21 - Ciências Exatas e da Terra, Sociais, da Saúde, Humanas e Engenharia/Tecnologia 11 11 v.2, n.7, 2021 RECIMA21 - REVISTA CIENTÍFICA MULTIDISCIPLINAR
ISSN 2675-6218 EDUCAÇÃO 4.0 E COMO A ENFERMAGEM SE INSERE:
APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas APLICABILIDADE DAS NOVAS TECNOLOGIAS
Leticia Falcão Nogueira, Emanuel Pereira dos Santos,
Claudiane Blanco Andrade dos Santos, Vera Lúcia Freitas PANIAGUA, A.; ISTANCE, D. Teachers as Designers of Learning Environments: the importance of
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10.33448/rsd-v10i10.18867. Disponível em: https://rsdjournal.org/index.php/rsd/article/view/18867. Acesso em: 10 aug. 2021. SCHWAB, K.; DAVIS, N. Shaping the future of the fourth industrial revolution. New York:
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móvel. 2019. 241p. Tese (doutorado em Educação) - Universidade Federal do Rio Grande do Sul,
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pandemia: uma análise da desigualdade de acesso à internet na periferia de Curitiba. Revista
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https://doi.org/10.12957/riae.2020.51097. Acesso em: 08 ago. 2021. UNESCO. United Nations Educational, Scientific and Cultural Organization. COVID19 Educational
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Disruption and Response. 2020. Disponível em: https://pt.unesco.org/covid19/educationresponse. Acesso em: 12 jul. 2021. 12 12
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Electronic friction and tuning on atomically thin MoS2
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INTRODUCTION adhesion between graphene and supporting SiO2/Si substrate
via plasma treatment of the substrate to suppress the puckering
effect16,22. However, the tuning of the friction on atomically thin
graphene is irreversible because the interfacial bonding is hard to
recover to the initial state23. The friction on monolayer graphene
can be modulated reversibly by tuning the atomic-level contact
quality with simple mechanical straining under the relatively small
range of applied load18. In a word, the energy dissipation of
friction on atomically thin 2D materials is still undefined for
controllable tuning. Friction exists in almost all mechanical systems with moving parts
and it accounts for about a third of the world’s energy
consumption1,2. Consequently, friction is the process of energy
dissipation when the surfaces of moving parts slide against each
other. The friction reduction is desirable at all scales from macro to
nano for saving energy3,4. Atomically thin 2D materials such as
graphene, transition metal dichalcogenides (e.g., MoS2), and
boron nitride (h-BN) exhibit excellent lubrication properties and
attract wide attention as solid lubricants in confined layers of
molecular thickness for various micro-and nanoelectromechanical
systems (MEMS/NEMS)5–8. Also, alternating hold and release the
interfacial parts for steady operation in the control knob and
sliding actuators of 2D materials-based MEMS/NEMS3 require
controllable friction. It is necessary to not only seek the
contributions but also the basic science such as energy dissipation
to be understood to the friction on 2D materials effectively. The
atomic force microscopy (AFM) methods have allowed basic
studies of friction mechanisms on atomically thin 2D materials
which intensively address how energy is dissipated at a sliding
contact9. g
The sliding AFM tip during friction involves the direct transfer of
energy into the phononic dissipation and electron-hole pairs
before transfer to the phonon populations. The electron-phonon
coupling (EPC) becomes another channel for energy dissipation
and builds a link between electronic and phononic effects12. Consequently, the energy of friction can be dissipated through
phononic consumption and electronic contribution24–26. Electro-
nic contributions to energy dissipation of friction on atomically
thin 2D materials need to be manifested in many physical
phenomena. Dong27 quantitatively calculated the effect of EPC on
the friction of monolayer graphene by MD simulation with a
Langevin equation and found the friction increases slightly with
the increase of EPC. www.nature.com/npj2dmaterials ARTICLE
OPEN
Electronic friction and tuning on atomically thin MoS2 Bin Shi
1, Xuehui Gan1,2, Kang Yu1, Haojie Lang1, Xing’an Cao1, Kun Zou1 and Yitian Peng
1,2✉ Friction is an energy dissipation process. However, the electronic contribution to energy dissipation channels remains elusive
during the sliding friction process. The friction and dissipation on atomically thin MoS2 with semiconductive characteristics are
studied and tuned by the gate-modulated carrier concentration. The electronic contribution to energy dissipation of friction on
atomically thin MoS2 was confirmed and regulated through tuning the strength of the electron-phonon coupling. The electron-
phonon coupling can be strengthened and depressed to increase and decrease friction by the gate-modulation of the carrier
concentration. The fitting of the friction on atomically thin MoS2 and carrier concentration is approximately linear which is in
accordance with Langevin equation induced friction. Then the active, dynamical, and repeated tuning of friction on atomically thin
MoS2 with semiconductive properties is achieved by the active modulation of carrier concentration with gate voltage. These
observations help us to understand the electronic friction in essence, provide a utility approach to tune the friction intelligently on
atomically thin two-dimensional materials with semiconductive properties and achieve superlubric properties for the application in
various micro-and nanoelectromechanical systems. npj 2D Materials and Applications (2022) 6:39 ; https://doi.org/10.1038/s41699-022-00316-6 1College of Mechanical Engineering, Donghua University, Shanghai 201620, China. 2Shanghai Collaborative Innovation Center for High Performance Fiber Composites, Donghua
University, Shanghai 201620, China. ✉email: yitianpeng@dhu.edu.cn College of Mechanical Engineering, Donghua University, Shanghai 201620, China. 2Shanghai Collaborative Innovation Center for High Pe
niversity, Shanghai 201620, China. ✉email: yitianpeng@dhu.edu.cn College of Mechanical Engineering, Donghua University, Shanghai 201620, China. 2Shanghai Collaborative Innovation Center for High Performance Fiber Composites, Donghua
University Shanghai 201620 China ✉email: yitianpeng@dhu edu cn hua University, Shanghai 201620, China. 2Shanghai Collaborative Innovation Center for High Performance Fiber Composites, Donghua
ail: yitianpeng@dhu.edu.cn INTRODUCTION Filleter et al.12 found that the nanofriction on
monolayer graphene was twice than that on bilayer graphene due
to the strong EPC in monolayer graphene by means of angle-
resolved photoemission spectroscopy. While a deep understand-
ing of the EPC affects the frictional properties of 2D materials in
essence is still required. Park et al.28 discovered that the
accumulation of carriers near the semiconducting surface leads
to excess friction due to high carrier density. Furthermore, the
electron doping for single-layer and bilayer graphene transistors
has been investigated with non-destructive Raman spectroscopy
characterization based on the renormalization of the G and 2D
modes as a function of carrier concentration29. However, the
electronic contribution to friction during sliding contacts provides There has been considerable progress in understanding the
energy dissipation of friction on atomically thin 2D materials in the
past few decades10. The structure11,12 and substrate13–15 affect the
friction on atomically thin 2D materials deposited on the substrate
due to the elastic deformation15, adhesion induced puckering16,
and contact quality17,18. The kinetic energy associated with the
AFM tip sliding on atomically thin 2D materials could attribute to
dissipation by coupling the atomic relaxations to phonon modes
that propagate away and the elastic deformation energy due to
the out of plane deformation16–18. Then the defect generation and
chemical functionalization of atomically thin 2D materials could
tune the friction by increasing locally the potential corrugation
and lowing the bending rigidity, but destroy the structure
permanently19–21. The friction on the surface of atomically thin
graphene
is
reduced
dramatically
by
enhancing
interfacial Published in partnership with FCT NOVA with the support of E-MRS B. Shi et al. Fig. 1
The nanofriction experiments under different Vg. a Schematic of the nanofriction measurements on atomically thin MoS2 unde
different Vg. b AFM image and optical image(inset) of atomically thin MoS2 including the height profile of atomically thin MoS2 along the re
dash line(inset). Scale bar: 2 μm. c Friction map with the applied load of 10 nN under the modulation of Vg from 0 V to −5 V and back to 0 V
The inset shows the profiles of the nanofriction indicated by the white arrow line. Scale bar: 100 nm. d Friction map with the applied load o
10 nN under the modulation of Vg from 0 V to 5 V, then back to 0 V. INTRODUCTION The inset shows the profiles of the nanofriction indicated by the whit
arrow line. Scale bar: 100 nm. e The nanofriction on atomically thin MoS2 as a function of applied load with different Vg. f The nanofriction at
constant load of 10 nN under different Vg. g The nanofriction on atomically thin MoS2 varied with the different Vg. Error bars represen
standard deviations. B. Shi et al. 2 Fig. 1
The nanofriction experiments under different Vg. a Schematic of the nanofriction measurements on atomically thin MoS2 under
different Vg. b AFM image and optical image(inset) of atomically thin MoS2 including the height profile of atomically thin MoS2 along the red
dash line(inset). Scale bar: 2 μm. c Friction map with the applied load of 10 nN under the modulation of Vg from 0 V to −5 V and back to 0 V. The inset shows the profiles of the nanofriction indicated by the white arrow line. Scale bar: 100 nm. d Friction map with the applied load of
10 nN under the modulation of Vg from 0 V to 5 V, then back to 0 V. The inset shows the profiles of the nanofriction indicated by the white
arrow line. Scale bar: 100 nm. e The nanofriction on atomically thin MoS2 as a function of applied load with different Vg. f The nanofriction at a
constant load of 10 nN under different Vg. g The nanofriction on atomically thin MoS2 varied with the different Vg. Error bars represent
standard deviations. a possible method to tune the friction on atomically thin 2D
materials through the carrier concentration30. over many orders of magnitude. Scanning Kelvin force microscopy
(SKPM) has been used to measure the contact potential difference
(CPD) of atomically thin MoS2 under the gate voltage to calculate
the variation of carrier concentration during the carrier accumula-
tion and depletion36,37. The strengthening degree of EPC on
atomically thin MoS2 could be detected by the Raman spectra. As
the degree of EPC correlated strongly with carrier concentration,
the potential distribution of atomically thin MoS2 provides the
connection of carrier concentration with energy dissipation based
on the modulation of gate voltage38. Then, the friction on
atomically thin MoS2 could be tuned by regulation of the carrier
concentration over many orders of magnitude based on field
effect. Published in partnership with FCT NOVA with the support of E-MRS npj 2D Materials and Applications (2022) 39 INTRODUCTION The semiconductor materials offer an interesting platform to
investigate the electronic contribution to energy dissipation
where energy dissipation is linked to interaction with the electron
system. The charge carrier concentration and energy band
alignment of semiconductors could be extensively and precisely
controlled by the electric field31. In contrast to the zero gaps of
graphene
and
wide
gap
of
boron
nitride32,
mechanically
exfoliated atomically thin MoS2 exhibits unique n-type semicon-
ducting properties and tunable bandgap33. The carrier concentra-
tion in atomically thin MoS2 can be modulated easily with high
sensitivity in a relatively large range from ~1013 to 1015 cm−2 by
many orders of magnitude using the gate voltage34,35. Atomically
thin MoS2 with semiconductive properties provides the possibility
to study the effect of free charge carriers in the energy dissipation
of friction because it is possible to reversibly change their density In this manuscript, the carrier concentration of atomically thin
MoS2 deposited on SiO2/Si substrate is controlled by gate voltage
(Vg) based on field effect. Then the electronic friction and energy
dissipation
on
atomically
thin
MoS2
are
investigated
by Fig. 2
Force-distance curves on atomically thin MoS2 and variation of contact area under different Vg. a Adhesion and electrostatic force
as a function of Vg. b Typical force-distance curves measured under the positive Vg. c Typical force-distance curves measured under the
negative Vg. d Plots of the friction force versus the applied load on atomically thin MoS2 at Vg = 0 V. e The nanofriction caused by electrostatic
force (fele) and additional effect (fextra). f Variation of the true contact area during AFM tip sliding on atomically thin MoS2 under different Vg. Error bars represent standard deviations. B. Shi et al. B. Shi et al. 3 Fig. 2
Force-distance curves on atomically thin MoS2 and variation of contact area under different Vg. a Adhesion and electrostatic force
as a function of Vg. b Typical force-distance curves measured under the positive Vg. c Typical force-distance curves measured under the
negative Vg. d Plots of the friction force versus the applied load on atomically thin MoS2 at Vg = 0 V. e The nanofriction caused by electrostatic
force (fele) and additional effect (fextra). f Variation of the true contact area during AFM tip sliding on atomically thin MoS2 under different Vg. Error bars represent standard deviations. INTRODUCTION modulating the degree of EPC with varying carrier concentration
based on atomic-scale stick-slip calculation, surface potential, and
in-situ Raman spectroscopy under the external electric field. The
friction on atomically thin MoS2 is tuned dynamically, repeatably,
and
controllably
based
on
the
modulation
of
the
carrier
concentration further. substrate was investigated using calibrated AFM under the gate
voltage, as schematically shown in Fig. 1a. As shown in Fig. 1c, the
Vg was sequentially applied in the order of 0–−5 V and then return
to 0 V. The nanofriction on atomically thin MoS2 decreases at Vg =
−5 V and then recovered fully to its initial state when Vg was
returned to 0 V. The nanofriction has little change under relatively
low negative Vg < −5 V (Supplementary Information Fig. 1). The
nanofriction on atomically thin MoS2 at Vg = −5 V decreases about
20% contrast to the initial value from Fig. 1c. The nanofriction on
atomically thin MoS2 is reduced obviously at the negative gate
voltage of −5 V and decreases slightly with the increase of the
negative bias when the gate voltage is beyond −5 V. However, the
nanofriction on atomically thin MoS2 under the Vg = 5 V increases
almost to 1.8 times than the initial value, as shown in Fig. 1d. The
nanofriction on atomically thin MoS2 increases with the increase Published in partnership with FCT NOVA with the support of E-MRS npj 2D Materials and Applications (2022) 39 Mechanism The adhesion on atomically thin MoS2 were measured using AFM
under different Vg. Figure 2a shows the adhesive force on
atomically thin MoS2 and electrostatic force as a function of the Vg. The adhesion Fad approximately follows a parabolic law, i.e., Fad ∝
V 2, with little difference between positive and negative Vg. The Fad
mainly consists of van der Waals force (FvdW), electrostatic force
(Fele), capillary force (Fcap), and chemical bonding force (Fchem) in
an ambient environment, which can be written as: (1) Fad ¼ FvdW þ Fele þ Fcap þ Fchem
(1) Fad ¼ FvdW þ Fele þ Fcap þ Fchem The atomic-scale stick-slip behaviors on atomically thin MoS2
were measured using AFM to comprehend the underlying
mechanism under different Vg further. The lateral force curves at
atomic scale under different Vg are shown in Fig. 3a. The sawtooth
shape of the trace shows the atomic lattice period of about
0.31 nm corresponding to the lattice constant of atomically thin
MoS2. It is also clearly observed that the distance of the stick-slip
period increases from 0.31 nm to 0.87 nm when the Vg was
regulated from 0 V to 4 V. The atomic-level stick-slip behaviors of
the lateral force on atomically thin MoS2 change with Vg, then
return to the original. As the Vg increased, the stick-slip behaviors
switch from single-slip to multiple-slip regime. The multiple-slip
regime can also be fully recovered to a single-slip regime when Vg
returns from 4 V to 0 V, as shown in Fig. 3a. The reversible
transition between single slip and multiple slips indicates that the
atomic-scale stick-slip behaviors can be reversibly modulated by
the Vg. The lateral force map in the sliding process under different
Vg is shown in Fig. 3b. Figure 3b was used to analyze the lateral
force map in the sliding process under different gate voltages. Figure 3b indicated that the multiple-slip regime can also be fully
recovered to a single-slip regime when Vg changes from 0 V to 6 V
and then return to 0 V. Few regular atomic stick-slip motions are
also observed when Vg ≥6 V (Supplementary Information Fig. 5). As shown in Fig. 3c, the mean lateral force increases with the
increase of Vg. In the meantime, the total energy dissipation
calculated from the atomic-scale stick-slip curves increased
monotonically with the increase of the positive Vg shown in
Fig. 3c. Mechanism In addition, the distance between the trace and retrace
values decreases demonstrating less energy dissipation for the
decrease of average nanofriction under the negative Vg. The surface properties of mechanically exfoliated atomically
thin MoS2 keep highly hydrophobic and stable with barely
dangling bonds under different Vg (Supplementary Information
Fig. 3). Also, any effect of a changing tip structure, geometrically or
chemically can be neglected during the nanofriction experiment
under different Vg. The change of FvdW, Fcap, and Fchem can be
neglected for the increase of the adhesion between the AFM tip
and atomically thin MoS2 under different Vg. Consequently, the
main reason for the change of adhesion force is the effect of Fele
under different Vg. The electrostatic force caused by the electric
field could be quantitatively calculated from the adhesion
measurement of the single force curve under different Vg. So
the Fele can be calculated using Eq. (1) at different Vg, as shown in
Fig. 2a. Based on the quadratic fit of the electrostatic force, the
relationship between the Fele and the Vg is (2) Fele ¼ 0:14ðVg 0:64Þ2 þ 0:07
(2) Fele ¼ 0:14ðVg 0:64Þ2 þ 0:07 The electrostatic forces at Vg = 0 V, 5 V, 10 V, −5 V and −10 V
are about 0 nN, 2.73 nN, 12.4 nN, 4.69 nN and 15.92 nN, respec-
tively. The electrostatic force would increase the interaction
between the tip and the atomically thin MoS2 and create
additional friction. Therefore, the additional nanofriction (fele)
caused by Fele was calculated while the Fele was treated as an
additional load to the AFM tip. The typical single force curves on
the atomically thin MoS2 were acquired with the AFM tip under
the positive Vg, as shown in Fig. 2b. The difference of pull-off force
between 0V and 6 V reveals the attraction contact because of the
electrostatic interaction. The typical single force curves on the
atomically thin MoS2 under the negative Vg shown in Fig. 2c have
a similar tendency as the positive Vg. g
The increase in the lateral force of stick-slip asymmetry suggests
an incremental energy corrugation (see Supplementary Informa-
tion Fig. 6)39. Therefore, a large lateral force is required to cross the
barrier resulting in more energy dissipation under the positive Vg. The lateral contact stiffness was extracted from the slope of each
stick phase in lateral force curves at the atomic scale in Fig. 3a. Friction properties under the electric field Consequently, there is
an additional mechanism for the increase on the nanofriction but
the load. Considering that the change of adhesion force is closely
related to the change of contact area, the nanofriction usually
agrees well with the true contact area based on the contact model
of Derjaguin-Müller-Toporov (DMT). As shown in Fig. 2f, the
changes of contact area between the AFM tip and atomically thin
MoS2 (Detailed calculation process in Supplementary Information
Fig. 4) could not account for the obvious variation in nanofriction
under different Vg. These findings indicate that the fele plays a
small role in the change of nanofriction on atomically thin MoS2
under the different Vg. Meanwhile, the gate voltage induced
another factor to change the nanofriction on atomically thin MoS2
besides the contact area. where f0 is the nanofriction at Vg = 0 V, fg is the nanofriction force
under the different Vg with the applied load of 7.8 nN. The force of
electrostatic attraction was supposed to increase the adhesion
force, then increase nanofriction under the normal electric field. Figure 2e also shows that the electrostatic force is insufficient to
increase the nanofriction to such an extent. Consequently, there is
an additional mechanism for the increase on the nanofriction but
the load. Considering that the change of adhesion force is closely
related to the change of contact area, the nanofriction usually
agrees well with the true contact area based on the contact model
of Derjaguin-Müller-Toporov (DMT). As shown in Fig. 2f, the
changes of contact area between the AFM tip and atomically thin
MoS2 (Detailed calculation process in Supplementary Information
Fig. 4) could not account for the obvious variation in nanofriction
under different Vg. These findings indicate that the fele plays a
small role in the change of nanofriction on atomically thin MoS2
under the different Vg. Meanwhile, the gate voltage induced
another factor to change the nanofriction on atomically thin MoS2
besides the contact area. Friction properties under the electric field The topography of the atomically thin MoS2 on SiO2/Si substrate
acquired by AFM in tapping mode is shown in Fig. 1b. The height
of atomically thin MoS2 is about 0.9 nm corresponding to a single
layer from the topography profile in the inset of Fig. 1b. Then, the
nanofriction of atomically thin MoS2 deposited on SiO2/Si npj 2D Materials and Applications (2022) 39 B. Shi et al. 4 of positive gate voltage from 0 to 5 V, then recovered fully to its
initial value when Vg was returned to 0 V, as shown in Fig. 1d. Therefore, the nanofriction on atomically thin MoS2 can be
reversibly tuned by the regulation of Vg. Also, the polarity of the
applied Vg exhibits a different effect on the nanofriction of
atomically thin MoS2. Figure 1e shows the curves of nanofriction
on atomically thin MoS2 versus the applied load measured under
different Vg. Figure 1f shows the nanofriction on atomically thin
MoS2 at a constant load of 10 nN under different Vg. The
corresponding relationship between the magnitude of nanofric-
tion and the applied Vg is shown in Fig. 1g. The high-resolution
AFM image of atomically thin MoS2 denotes that it can remain
stable after the friction test under the external electric field
(Supplementary Information Fig. 2). It can be clearly seen that the
nanofriction on atomically thin MoS2 was affected by the direction
and magnitude of the Vg. 0.19 nN, and 0.64 nN at the Vg of 0 V, 5 V, 10 V, −5 V, and −10 V,
respectively. Consequently, the increases of the nanofriction
under different Vg are far more than the additional fele caused
by electrostatic force. It can be concluded that there is an
additional effect leading to the nanofriction increasing. Therefore,
the nanofriction (fextra) caused by the additional action can be
given by the equation: fextra ¼ fg fele f0
(4) fextra ¼ fg fele f0 (4) where f0 is the nanofriction at Vg = 0 V, fg is the nanofriction force
under the different Vg with the applied load of 7.8 nN. The force of
electrostatic attraction was supposed to increase the adhesion
force, then increase nanofriction under the normal electric field. Figure 2e also shows that the electrostatic force is insufficient to
increase the nanofriction to such an extent. Published in partnership with FCT NOVA with the support of E-MRS npj 2D Materials and Applications (2022) 39 Mechanism It
has been reported that the value of lateral contact stiffness
increases with load, indicating the increase of the contact area40. However, Fig. 3d shows that the lateral contact stiffness is almost
kept stable under different Vg at a given load. Generally, the lateral y
p
g
The initial value of nanofriction starts with the adhesion force of
7.8 nN and the corresponding normal load is also 7.8 nN. Therefore, fele can be given by (3) fele ¼ μ ´ Fele where μ is defined as the equal friction coefficient fitted from the
friction as a function of load, as shown in Fig. 2d. The values of fele
were calculated approximately to be 0 nN, 0.11 nN, 0.50 nN, Published in partnership with FCT NOVA with the support of E-MRS B. Shi et al. Fig. 3
Atomic-scale friction and stick-slip behavior on atomically thin MoS2 under different Vg. a Stick-slip behavior measured at Vg = −4–
+4 V, respectively. b Lateral force mapping under different Vg: the gate bias voltage was dynamically modulated from 0 V to 6 V and return to
0 V. Scale bar: 1 nm. c Energy dissipation and lateral peak force under different Vg. d The lateral contact stiffness k under different Vg. Error bars
represent standard deviations. 5 Fig. 3
Atomic-scale friction and stick-slip behavior on atomically thin MoS2 under different Vg. a Stick-slip behavior measured at Vg = −4–
+4 V, respectively. b Lateral force mapping under different Vg: the gate bias voltage was dynamically modulated from 0 V to 6 V and return to
0 V. Scale bar: 1 nm. c Energy dissipation and lateral peak force under different Vg. d The lateral contact stiffness k under different Vg. Error bars
represent standard deviations. trivial effect of the Vg is to modulate the carrier concentration
based on the electric field effect42. The carrier concentration
can be calculated based on n ¼ Ncexp
ECEF
ð
Þ
KBT
43,44, where
Nc = (2m*)/(πħ2) = 3.8 × 1014eV cm−2 is the density of states in
the conduction band, EC is the bottom of the conduction band, KB
is Boltzmann constant (KB = 1.380649 × 10−23 J/K), T is the
temperature (300 K). EF is the Fermi energy level. EF can be
calculated by measuring the surface potential through SKPM. Therefore, the SKPM test is used to calculate the carrier
concentration of MoS2. Published in partnership with FCT NOVA with the support of E-MRS npj 2D Materials and Applications (2022) 39 Mechanism The work function of MoS2 can be
calculated by SKPM based on the formula of ΔCPD = (Wtip −
Wsample)/e45, where Wsample and Wtip are the work functions of the
MoS2 and tip, e is the electronic charge. The work function of
MoS2 can be calculated by measuring ΔCPD with SKPM, and then
the Fermi energy level of MoS2 can be calculated by W = 0−EF. However, the surface potential of MoS2 is not only determined by
the work function of MoS2 but also directly affected by the electric
field after applying the gate voltage. The surface potential of SiO2
under the different Vg was measured and the results are shown in contact stiffness would show a prominent deviation by the sliding
energy barrier41. In addition, the average shear stress of the
contact was analyzed. The friction (F) is proportional to the real
contact area (A) as F = τ × A41, where τ is the shear strength. The
real contact area under different gate voltages has been obtained
based on the DMT model shown in Fig. 2f. The variation of the
contact area and shear strength with gate voltages is shown in
Supplementary Fig. 7 (see Supplementary Information). The shear
strength increases under the positive gate voltages and decreases
under the negative gate voltages. However, the data in Fig. 3
shows that the contact stiffness remains unchanged under
different bias voltages. The reason is that Formula (1) holds if
electronic contributions to friction are not involved28. These
results indicated that the shear stress is hard to explain the
experimental results of MoS2 under the electric field. Conse-
quently, the variation of nanofriction on atomically thin MoS2 at
different Vg can attribute to another factor beyond the energy
dissipation from the energy barrier. In addition to the changes in electrostatic force between the tip
and atomically thin MoS2 under the external electric field, a more Published in partnership with FCT NOVA with the support of E-MRS npj 2D Materials and Applications (2022) 39 B. Shi et al. Fig. 4
Surface potential mapping of atomically thin MoS2 on SiO2/Si substrate. a–l The surface potential mapping of atomically thin MoS2
under different Vg: −6–+6 V. Scale bar: 2 μm. m The line profiles of surface potential along the white line across the atomically thin MoS2
under different Vg. n ΔCPD of atomically thin MoS2 as a function of Vg. npj 2D Materials and Applications (2022) 39 Mechanism Therefore, the carrier concentration was calculated based
on n ¼ Ncexp
ECEF
ð
Þ
KBT
, where EC = −3.9246, KBT = 0.0259 eV47,
E
W
Th
k f
i
f
d
i
i
i
V Th of MoS2 is lower than the potential of SiO2 under the positive gate
voltages indicating that the MoS2 is an n-type semiconductor,
while the potential of MoS2 is higher than the potential of SiO2
under the negative gate voltages indicating that the n region is
reverse-biased causing depletion or weak inversion36. Surface
potential profiles along the atomically thin MoS2 on the SiO2/Si
substrate under the different Vg are shown in Fig. 4m. The values
of ΔCPD are given in Fig. 4n after applied gate voltage from −6
to +6 V. Therefore, the carrier concentration was calculated based
on n ¼ Ncexp
ECEF
ð
Þ
KBT
, where EC = −3.9246, KBT = 0.0259 eV47,
E
W
Th
k f
i
f
d
i
i
i
V Th of MoS2 is lower than the potential of SiO2 under the positive gate
voltages indicating that the MoS2 is an n-type semiconductor,
while the potential of MoS2 is higher than the potential of SiO2
under the negative gate voltages indicating that the n region is
reverse-biased causing depletion or weak inversion36. Surface
potential profiles along the atomically thin MoS2 on the SiO2/Si
substrate under the different Vg are shown in Fig. 4m. The values
of ΔCPD are given in Fig. 4n after applied gate voltage from −6
to +6 V. Therefore, the carrier concentration was calculated based
on n ¼ Ncexp
ECEF
ð
Þ
KBT
, where EC = −3.9246, KBT = 0.0259 eV47,
EF = −Wsample. The work function of conductive tip is 4.29 eV. The
ΔCPD is 0.043 V at the gate voltage of 0V and the corresponding
Fermi energy level is EF = −4.22. The ΔCPD is 0.102 V at the gate
voltage of 5 V and the corresponding Fermi energy level is EF =
−4.13. Published in partnership with FCT NOVA with the support of E-MRS Mechanism Error bars represent standard deviations. 6 Fig. 4
Surface potential mapping of atomically thin MoS2 on SiO2/Si substrate. a–l The surface potential mapping of atomically thin MoS2
under different Vg: −6–+6 V. Scale bar: 2 μm. m The line profiles of surface potential along the white line across the atomically thin MoS2
under different Vg. n ΔCPD of atomically thin MoS2 as a function of Vg. Error bars represent standard deviations. Supplementary Fig. 8 (Supplementary Information). To minimize
the influence of the applied electric field on the SKPM test, the
surface potential of SiO2 was used as the reference. Because the
relationship between the surface potential of SiO2 and gate
voltage is linear. The surface potential of SiO2 is consistent with
the Vg indicating that the work function of SiO2 as an insulator is
very little affected by the Vg. Therefore, the surface potential
difference between SiO2 and MoS2 (ΔCPD) was used to reflect the
effect of electric field on MoS2, which is relatively more accurate. The surface potential of MoS2 deposited on SiO2/Si substrate
under different Vg was shown in Fig. 4a–l. The yellow dotted box
denotes the area of the atomically thin MoS2 on the surrounding
SiO2/Si substrate. The surface potential of atomically thin MoS2
increases with the increase of positive Vg as presented in Fig. 4a–f. Also, the surface potential of atomically thin MoS2 decreases with
a decrease of the negative Vg, as shown in Fig. 4h–l. The potential of MoS2 is lower than the potential of SiO2 under the positive gate
voltages indicating that the MoS2 is an n-type semiconductor,
while the potential of MoS2 is higher than the potential of SiO2
under the negative gate voltages indicating that the n region is
reverse-biased causing depletion or weak inversion36. Surface
potential profiles along the atomically thin MoS2 on the SiO2/Si
substrate under the different Vg are shown in Fig. 4m. The values
of ΔCPD are given in Fig. 4n after applied gate voltage from −6
to +6 V. Mechanism The FWHM of Raman mode derives from three factors as
given below: ΓFWHM = Γ0 + Γanh + ΓEPC, where Γ0 is the intrinsic
FWHM, Γanh denotes the phonon-phonon interaction, and ΓEPC is
due to EPC51. Therefore, the ΓEPC can be used to illustrate the
strength degree of EPC. The Γ0 and Γanh are expected to be
the same at different Vg. The FWHM of A1g mode increases with
the increase of carrier concentration. However, the FWHM of the
E12g phonon was independent of the electron concentration38. The high level of carrier concentration causes the strong strength
of EPC from the change in the A1g. The ΓEPC increases with the
increased carrier concentration indicating that the strength of
EPC increases with the increased carrier concentration. The
friction increases with the increase of the carrier concentration, cm−2 at the gate voltage of 0 V (intrinsic carrier concentration)
and
0.12 × 1012 cm−2
at
the
gate
voltage
of
5 V. Carrier
concentration n can also be approximatively calculated using
the idealized parallel-plate capacitor model with n ¼ CSiO2 ´ Vg
(CSiO2 is ~15 nF cm−2 for 300 nm SiO2 dielectrics)48. Therefore, the
alternate surface potential of atomically thin MoS2 suggests that
the carriers accumulate under the positive Vg and dissipate under
the negative Vg from the surface potential characterization and
calculation. The variation of the CPD also indicates the direction and
amount of energy band bending and the position of the Fermi
level at the surface caused by surface electron states49. Since the
atomically thin MoS2 is n-type semiconductor, the Fermi level in
equilibrium is close to the conduction band (CB), as shown in
Fig. 5a. The energy band bends downward and Fermi level is
closer to the conduction band under the positive Vg. When a
positive Vg is applied, only a few tenths of electron-volts shift at
the conduction band edge50. This effect is clearly observed in
Fig. 4n, where a positive Vg results in a little increase of △CPD due
to its proximity to CB. However, the change of Fermi level under
the negative is significantly changed due to the large distance
between the Fermi level and the valence band (VB), as shown in
Fig. 5c. Therefore, a large increase in ΔCPD was observed under
the negative Vg. Furthermore, the variation in carrier concentration
contributes to the shift of the position of Fermi level by the band
bending. Mechanism Therefore, the carrier concentration of MoS2 is 3.4 × 109 of MoS2 is lower than the potential of SiO2 under the positive gate
voltages indicating that the MoS2 is an n-type semiconductor,
while the potential of MoS2 is higher than the potential of SiO2
under the negative gate voltages indicating that the n region is
reverse-biased causing depletion or weak inversion36. Surface
potential profiles along the atomically thin MoS2 on the SiO2/Si
substrate under the different Vg are shown in Fig. 4m. The values
of ΔCPD are given in Fig. 4n after applied gate voltage from −6
to +6 V. Therefore, the carrier concentration was calculated based
on n ¼ Ncexp
ECEF
ð
Þ
KBT
, where EC = −3.9246, KBT = 0.0259 eV47,
EF = −Wsample. The work function of conductive tip is 4.29 eV. The
ΔCPD is 0.043 V at the gate voltage of 0V and the corresponding
Fermi energy level is EF = −4.22. The ΔCPD is 0.102 V at the gate
voltage of 5 V and the corresponding Fermi energy level is EF =
−4.13. Therefore, the carrier concentration of MoS2 is 3.4 × 109
EF = −Wsample. The work function of conductive tip is 4.29 eV. The
ΔCPD is 0.043 V at the gate voltage of 0V and the corresponding
Fermi energy level is EF = −4.22. The ΔCPD is 0.102 V at the gate
voltage of 5 V and the corresponding Fermi energy level is EF =
−4.13. Therefore, the carrier concentration of MoS2 is 3.4 × 109 Published in partnership with FCT NOVA with the support of E-MRS Fig. 5
Energy band diagram of atomically thin MoS2 with n-type semiconductor properties. a Energy band diagrams of atomically thin
MoS2 at equilibrium. b The energy band bending and electron accumulation caused by the existence of donor-like surface states under the
positive gate voltage. The band diagram of the MoS2/SiO2/Si structure. c The band diagram of the MoS2/SiO2/Si structure at negative gate
voltage. The energy band bending and surface electron depletion under the negative gate voltage. B. Shi et al. B. Shi et al. B. Shi et al. 7 Fig. 5
Energy band diagram of atomically thin MoS2 with n-type semiconductor properties. a Energy band diagrams of atomically thin
MoS2 at equilibrium. Mechanism b The energy band bending and electron accumulation caused by the existence of donor-like surface states under the
positive gate voltage. The band diagram of the MoS2/SiO2/Si structure. c The band diagram of the MoS2/SiO2/Si structure at negative gate
voltage. The energy band bending and surface electron depletion under the negative gate voltage. atomically thin MoS2 was depicted in Fig. 6a. Figure 6b displays
the evolution of zone-center phonon E12g and A1g modes of the
atomically thin MoS2 under different Vg. As depicted in the inset
of Fig. 6b, the A1g phonon involves the sulfur atomic vibration in
the opposite direction along the c axis (perpendicular to the
basal plane), whereas the E12g mode illustrates the displacement
of Mo and S atoms are in the basal plane. The dependence of the
change in different renormalization mode frequencies of the two
modes A1g and E12g on the carrier concentration from the
obtained Line-shape parameters show that the A1g mode
frequency softens by 4 cm−1, as compared to only ~0.6 cm−1
for the E12g mode under the maximum voltage of 20 V. The
linewidth of the A1g mode increases significantly by ~0.98 cm−1
for the maximum doping achieved, whereas the linewidth of the
E12g mode does not show appreciable change. The renormaliza-
tion A1g mode could be used as a sign of the quantitative EPC
corresponding to the carrier concentration modulated by the Vg. The down-shift of A1g mode is due to the increase of carrier
concentration leading to the occupation of an anti-bonding state
in the conduction band of atomically thin MoS2. The occupation
of an anti-bonding state increases the total electron energy of
the system and weakens the Mo-S bond due to electron doping. Therefore, electron doping caused a significant change in the
EPC of the A1g mode, while the E12g mode of Raman spectra with
symmetry is quite insensitive to the external electric field. The
phonon frequency and linewidth of the E12g mode are much less
dependent on carrier concentration. Figure 6c, d show the shift
of the mode frequencies and the corresponding full width at half
maximum (FWHM) as a function of Vg, respectively. The A1g
mode specifically exhibits a strong sensitivity to electron doping
and the phonon frequency decreases by 4 cm−1 and the
linewidth broadens by 1.1 cm−1 for electron doping at the Vg
of 20 V. Published in partnership with FCT NOVA with the support of E-MRS npj 2D Materials and Applications (2022) 39 Mechanism The energy band of atomically thin MoS2 bends
downward and the Fermi level is closer to the conduction band
resulting in carrier accumulation under a positive Vg. Figure 5c
shows that the energy band bends upward and the Fermi level is
closer to the valence band under the negative Vg, indicating that
the carrier dissipates near the atomically thin MoS2 surface. Although the surface potential decreases significantly under the
negative Vg, the corresponding carrier concentration decreases
slightly. Similarly, the friction (or peak force) in Fig. 3c also shows a
slight decrease as the voltage increases from 0 V to −4 V. Therefore, the decrease of ΔCPD in Fig. 4n indicates that the
carrier concentration decreases under the negative gate voltage. Therefore, the decrease of carrier concentration under the
negative Vg may be closely related to the decrease of nanofriction. The nondestructive characterization of in-situ Raman spectro-
scopy
was
explored
to
investigate
the
effect
of
carrier
concentration on the EPC on atomically thin MoS2 under
different Vg. The schematic diagram of Raman spectroscopy
analysis
on
the
gate-modulated
carrier
concentration
of g
The nondestructive characterization of in-situ Raman spectro-
scopy
was
explored
to
investigate
the
effect
of
carrier
concentration on the EPC on atomically thin MoS2 under
different Vg. The schematic diagram of Raman spectroscopy
analysis
on
the
gate-modulated
carrier
concentration
of npj 2D Materials and Applications (2022) 39 Published in partnership with FCT NOVA with the support of E-MRS Fig. 6
The influence of normal electric field on nanofriction using Raman spectroscopy. a Raman experimental setup. b Raman spect
atomically thin MoS2 under different Vg. c FWHM and carrier concentration of A1g mode as a function of Vg. d FWHM and carrier concentra
of E12g mode as a function of Vg. e Carrier concentration and friction as a function of gate voltage. f Plots of the friction as a function of ca
concentration. g AFM tip sliding on atomically thin MoS2 emitting electron and phonon pairs. h Schematic diagram of the energy dissipa
as the AFM tip slides on atomically thin MoS2. Error bars represent standard deviations. B. Shi et al. 8 Fig. 6
The influence of normal electric field on nanofriction using Raman spectroscopy. a Raman experimental setup. b Raman spectra of
atomically thin MoS2 under different Vg. c FWHM and carrier concentration of A1g mode as a function of Vg. Mechanism ζi(t) is a random fluctuating force
and subject to the fluctuation-dissipation theorem53. The random
force ζi(t) can be solved with the equation of <ξ(t)ξ(t')≥2miRikBTe δ
(t −t'), where δ is the Dirac delta function. Ri can be given as
follows: (10) If α and β are equal to 0.97 and 1.23, respectively. Then, the
friction is linear with the carrier concentration from the Langevin
equation which is in accordance with the fitting of friction and
carrier concentration. Ri ¼ migep
3NkB
;
(7) Ri ¼ migep
3NkB
; (7) The strength of electron-phonon coupling in atomically thin
MoS2 increases with the increase of gate voltage due to electron
doping. Therefore, this lead us to the most likely scenario to
explain the increased nanofriction of atomically thin MoS2 under
the applied gate voltage. Phonons excited by the mechanical
energy of the sliding tip in atomically thin MoS2 are much more
likely to scatter with electrons, introducing a more efficient means
of dissipating energy12. During the observed stick-slip motion, the
lattice is locally distorted and released by the slipping probe tip
transferring kinetic energy into lattice vibrations26. For the
atomically thin MoS2 under the electric field, the lattice motion
is damped by the creation of electronic excitations through
electron-phonon coupling. This additional mechanism increases
the efficiency of energy dissipation during slip events which are
responsible for frictional energy losses. N is the number of atoms. ζi is related to gep and gep is the EPC
parameter54, which is gep ¼ π2meC2
s ne
6τ Ti
ð ÞTe , where me is the effective
electron mass, Cs is the speed of sound, ne is the electrons density,
and τ(Te) is the electron relaxation time defined as the electron-
phonon scattering time. The EPC parameter is proportional to the
electron density. Based on the equation of (7), the friction can be
given as, Ri ¼
mimeπ2C2
s
18k2
BNτ Te
ð
ÞTe
ne
(8) (8) The total friction Ftotal could be divided into two parts including
the electronic contribution and phononic consumption. The
friction caused by phononic consumption Fph could be described Fig. 7
Modified Prandtl−Tomlinson model and calculating of the energy corrugation. a Schematic of the modified Prandtl−Tomlinson
model describing that the AFM tip moves on the monolayer MoS2. b Energy corrugation Utot as a function of gate voltage. c Energy
corrugation UEPC of electronic friction as a function of gate voltage. Mechanism d FWHM and carrier concentration
of E12g mode as a function of Vg. e Carrier concentration and friction as a function of gate voltage. f Plots of the friction as a function of carrier
concentration. g AFM tip sliding on atomically thin MoS2 emitting electron and phonon pairs. h Schematic diagram of the energy dissipation
as the AFM tip slides on atomically thin MoS2. Error bars represent standard deviations. Fig. 6
The influence of normal electric field on nanofriction using Raman spectroscopy. a Raman experimental setup. b Raman spectra of
atomically thin MoS2 under different Vg. c FWHM and carrier concentration of A1g mode as a function of Vg. d FWHM and carrier concentration
of E12g mode as a function of Vg. e Carrier concentration and friction as a function of gate voltage. f Plots of the friction as a function of carrier
concentration. g AFM tip sliding on atomically thin MoS2 emitting electron and phonon pairs. h Schematic diagram of the energy dissipation
as the AFM tip slides on atomically thin MoS2. Error bars represent standard deviations. indicating that the energy dissipation of friction is consistent
with the strengthening of EPC. Both the friction and carrier
concentration
increase with the increase
of gate
voltage
confirming a strong correlation between the friction and carrier
concentration from Fig. 6e. The data of friction and carrier
concentration were fitted to quantify the relationship between
the friction and carrier concentration. The fitted curve in Fig. 6f
shows that the relationship between the friction (F) and carrier
concentration (ne) is Therefore, the friction to the carrier concentration approximately
follows a linear function of one variable. Figure 6g shows the schematic diagram of the energy
dissipation as the AFM tip slides on atomically thin MoS2
emitting phonon. The phonons produced by friction on
atomically thin MoS2 interact with electrons and lead to an
additional channel of energy dissipation by EPC, as schemati-
cally shown in Fig. 6h. Therefore, the strength of EPC in
atomically thin MoS2 was significantly changed by the gate-
modulated carrier concentration. The physical picture for the
change in nanofriction on atomically thin MoS2 under different F ¼ 0:97ne þ 1:23 (5) F ¼ 0:97ne þ 1:23 Published in partnership with FCT NOVA with the support of E-MRS npj 2D Materials and Applications (2022) 39 B. Shi et al. Mechanism 9 as Equation: Fph ¼ πA
b2ω E0 þ kTln
ν
ν0
h
i
55. Thus,
Ftotal ¼ FEPC þ Fph ¼
mimeπ2C2
s
18k2
BNτ Te
ð
ÞTe
ne ´ νi þ πA
b2ω E0 þ kTln
ν
ν0
(9) Vg was proposed: the instability of the stick-slip behavior would
cause local vibration of the crystal lattice and the vibration
energy is dissipated by the excitation and propagation of
phonons. However, the associated lattice motion is damped by
the creation of electronic excitations through EPC in atomically
thin MoS2 with high electron doping. Also, the EPC could
change the dynamic behavior of lattice motion. The trajectory
of a nucleus in atomically thin MoS2 during the friction process
follows a Langevin equation25,26. Therefore, the generalization
of the Langevin equation can be used to demonstrate the
energy dissipation due to EPC with a friction term Ri and a
random force ζi25,52, (9) The
mimeπ2C2
s
18k2
BNτ Te
ð
ÞTe ´ νi remains the same a can be treated as a
constant α because only the carrier concentration was modulated
by the electric field and other parameters such as temperature
and velocity were kept the same. Furthermore, the friction caused
by phononic consumption also remains the same. Thus, the Fph is
also set as a constant β ¼ πA
b2ω E0 þ kTln
ν
ν0
h
i
. Then the general
formula of friction on atomically thin MoS2 under the electric field
could be described as follows, mi
∂vi
∂t ¼ FiðtÞ
X
RiðtÞνi þ ζiðtÞ
(6) (6) Ftotal ¼
mimeπ2C2
s
18k2
BNτ Te
ð
ÞTe
νi ´ ne þ πA
b2ω E0 þ kTln
ν
ν0
¼ α ne þ β where mi is the atom mass, vi is the velocity, Fi(t) denotes the total
force exerted on atom (mean force), and Ri is the friction term
representing the strength of EPC. ζi(t) is a random fluctuating force
and subject to the fluctuation-dissipation theorem53. The random
force ζi(t) can be solved with the equation of <ξ(t)ξ(t')≥2miRikBTe δ
(t −t'), where δ is the Dirac delta function. Ri can be given as
follows: where mi is the atom mass, vi is the velocity, Fi(t) denotes the total
force exerted on atom (mean force), and Ri is the friction term
representing the strength of EPC. Published in partnership with FCT NOVA with the support of E-MRS Mechanism Error bars represent standard deviations. Fig. 7
Modified Prandtl−Tomlinson model and calculating of the energy corrugation. a Schematic of the modified Prandtl−Tomlinson
model describing that the AFM tip moves on the monolayer MoS2. b Energy corrugation Utot as a function of gate voltage. c Energy
corrugation UEPC of electronic friction as a function of gate voltage. Error bars represent standard deviations. Published in partnership with FCT NOVA with the support of E-MRS
npj 2D Materials and Applications (2022) 39 Published in partnership with FCT NOVA with the support of E-MRS npj 2D Materials and Applications (2022) 39 B. Shi et al. 10 The atomic-scale stick-slip curves under different gate voltages
are shown in Fig. 3a. The Prandtl–Tomlinson model has been
greatly accepted for depicting the universal stick-slip phenomena. Since only the carrier concentration is changed, the phononic
friction is a constant under different gate voltages. We assume
that a linear superposition of the phononic energy dissipation and
an independent electronic friction channel caused by electron-
phonon coupling (EPC)24,55. Then, the total energy corrugation can
be given by Utot = Uph + UEPC, where Uph is the energy corrugation
of phononic friction, UEPC is the energy corrugation of electric
friction induced by EPC. dissipation of friction. Therefore, the energy corrugation at −4 V is
approximated as the Uph. For a better analysis of the electronic
friction branch, we substract the Uph curve from the total Utot. What remains is the electronic friction contribution as a function
of gate voltage in Fig. 7c. By fitting the UEPC as a linear function of
the gate voltage, we find that the relationship between the energy
corrugation of UEPC and the gate voltage is linear, which is
consistent with the relationship between carrier concentration
and friction. The friction tuning effect induced by an electric current was
investigated using conductive atomic force microscopy and DFT
calculation by Song et al.58. DFT calculations propose that a larger
electron density fluctuation during sliding tends to have a larger
sliding barrier change resulting in a larger change of friction,
which confirmed the contribution of electron density fluctuation
to friction. The carrier concentration of MoS2 was tuned based on
field effect modulation. Increasing carrier concentrations would
increase the electron density fluctuation resulting in larger friction. Wang et al. Mechanism has performed friction experiments of a single-asperity
sliding on a high-Tc superconductor from 40 to 300 kelvin and
they found that the electronic friction exceeds 50% of the total
friction signal24. Friction on single-layer graphene is found to be a
factor of two larger than on bilayer films due to the electron-
phonon coupling12. In our manuscript, the phonon dissipation is
governed by the energy dissipation of friction as the gate voltage
is below 0V due to electron dissipation. The friction could be
reduced by 17% under the negative voltage. Also, the nanofriction
does not decrease further when the gate voltage exceeds −5 V. As
a result, we infer that the electronic friction at the gate voltage of
0V is ~17% and phononic friction 83%. In addition, the charge
carriers
accumulate
and
the
electron-phonon
coupling
is
enhanced under the positive gate voltage, leading to more
efficient energy dissipation due to electron doping. As a result, the
electronic contribution dominates the energy dissipation in y
Therefore, a modified Prandtl−Tomlinson model was proposed,
as shown in Fig. 7a. An AFM tip moves on MoS2 surface can be
simplified to the motion of a point mass being dragged over a
periodic potential by a supporting body M through a spring. The
elastic spring of stiffness k represents the torsional stiffness of the
AFM cantilever. The system is characterized by the following
potential energy56,57: V ¼ Uph þ UEPC
2
cos 2π xtip
a
þ 1
2 k xtip xs
2
(11) (11) According to the modified PT model, the maximum lateral force
max (Fmax L) during the stick-slip motion is determined by the
interface energy corrugation Uph and UEPC. The maximum of the
absolute value of the force Fmax L is found at xtip = a/4, and we
obtain Utot ¼ a ´ Fmax
L
=π
(12) (12) Utot ¼ a ´ Fmax
L
=π The Utot was shown in Fig. 7b. The Utot increases significantly
under the positive gate voltage and decreases slightly under the
negative gate voltage. When the gate voltage is below 0V, the
electronic friction was suppressed due to the dissipation of charge
carriers and the phonon dissipation is governed by the energy Fig. 8
Nanofriction modulation on atomically thin MoS2. a Mapping of the nanofriction on atomically thin MoS2 modulated dynamically
under different Vg. Scale bar: 100 nm. npj 2D Materials and Applications (2022) 39 METHODS 11. Fajardo, O. Y. & Mazo, J. J. Surface defects and temperature on atomic friction. J. Phys. Condens. Mat. 23, 355008 (2011). DATA AVAILABILITY The data that support the plots within this paper and other findings of this study are
available from the corresponding author upon reasonable request. The data that support the plots within this paper and other findings of this study are
available from the corresponding author upon reasonable request. The data that support the plots within this paper and other findings of this study are
available from the corresponding author upon reasonable request. Received: 23 September 2021; Accepted: 16 May 2022; Raman spectroscopy characterization of atomically thin MoS2 Room-temperature Raman spectra were recorded with 532-nm line of an
argon ion laser as exciting radiation with a Witec confocal spectrometer
using a 50 × long working distance objective. Laser power was kept below
1 mW to avoid heating samples. The positive and negative gate voltages
were also applied to the conductive gold foil on the SiO2/Si substrate
during Raman measurements. Topography, friction, and adhesion measurements Topography, friction, and adhesion measurements The topography and thickness of atomically thin MoS2 were determined
by AFM in tapping mode (MFP-3D, Asylum Research Inc). The nanofriction
and adhesion measurements were accomplished on atomically thin MoS2
using AFM in contact mode in an ambient environment (temperature
~22 °C, relative humidity ~30%). The nanofriction on atomically thin MoS2
was measured in the scanning area of 500 × 500 nm2 at the scanning
speed of 1.25 μm/s. The adhesions were evaluated by the pull-off force
from force-distance measurements with a Si probe. Adhesion force was
measured five times to take the average value. The SiO2/Si substrate was
placed on the conductive gold foil for the connection of the external
electric field. The positive and negative values of the gate voltages (Vg)
were applied to the conductive gold foil during the nanofriction and
adhesion measurements. g
g
The nanofriction induced phonons in atomically thin MoS2 interact
strongly with electrons due to EPC resulting in a new channel for
energy dissipation of friction. The nanofriction on atomically thin
MoS2 increases with the increase of the carrier concentration,
indicating that the energy dissipation of friction is consistent with the
strengthening of EPC. The background carrier concentration was
reduced by applying the negative Vg. Therefore, the decreased carrier
concentration depresses the strength of the EPC to decrease the
additional energy dissipation efficiency in atomically thin MoS2. Then,
the nanofriction on atomically thin MoS2 correlates strongly with the
carrier concentration. Therefore, active controlling of nanofriction on
atomically thin MoS2 could be obtained by tuning the carrier
concentration based on gate voltage modulation. REFERENCES 1. Chu, S. & Majumdar, A. Opportunities and challenges for a sustainable energy
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nanotribology. J. Phys. D. Appl. Phys. 41, 123001 (2008). nanotribology. J. Phys. D. Appl. Phys. 41, 123001 (2008). 10. Andersson, D. & de Wijn, A. S. Understanding the friction of atomically thin
layered materials. Nat. Commun. 11, 420 (2020). Surface potential measurements The surface potential of atomically thin MoS2 was measured by scanning
Kelvin force microscopy (SKPM) under an ambient condition with different
gate voltages. The spring constant and resonance frequency of the
conductive probe for surface potential measurements was 2.8 N/m and
75 kHz, respectively. g
g
Figure. 8a shows 500 × 500 nm2 frictional map (grayscale:
bright, high nanofriction; dark, low nanofriction) on the atomically
thin MoS2 under the cyclic Vg of 5 V. The Vg was sequentially and
dynamically modulated in the order of 0-5-0-5-0-5-0-5-0-5 V. A
strip of nanofriction was observed from nanofriction map under
the applied electric field. As shown in Fig. 8b, it is clearly observed
that the nanofriction increases as the increase of Vg. Also, the
changes of nanofriction on atomically thin MoS2 are rapid and
reversible during the switch of the Vg. Figure. 8c shows the
frictional profiles during the dynamic tuning of Vg between −10 V
and 0 V. The nanofriction is reduced reversibly at Vg = −10 V and
the nanofriction can also be fully recovered when the Vg return to
0 V, as shown in Fig. 8c. Figure. 8d shows that the nanofriction is
controlled stably and reversibly at high Vg, proving the high
reliability of this nanofriction-tuning method. Figuer. 8d also
shows that the nanofriction response has good repeatability and
stability to the variation of Vg. The nanofriction can reduce by
about 20% under the negative Vg (Fig. 8c) and increase about five
times under the positive Vg (Fig. 8d). In particular, the effect of the
negative and positive Vg on the nanofriction have a significant
difference. Therefore, actively tuning of nanofriction on atomically
thin MoS2 in a dynamic, reversible, and controllable way is
achieved based on the field effect modulation of the carrier. Mechanism b Corresponding profiles of the nanofriction indicated by the blue arrow line. c The nanofriction on
atomically thin MoS2 modulated by applying the negative Vg. d The nanofriction on atomically thin MoS2 modulated by applying different
positive Vg (the load for friction test was 10 nN). Fig. 8
Nanofriction modulation on atomically thin MoS2. a Mapping of the nanofriction on atomically thin MoS2 modulated dynamically
under different Vg. Scale bar: 100 nm. b Corresponding profiles of the nanofriction indicated by the blue arrow line. c The nanofriction on
atomically thin MoS2 modulated by applying the negative Vg. d The nanofriction on atomically thin MoS2 modulated by applying different
positive Vg (the load for friction test was 10 nN). Published in partnership with FCT NOVA with the support of E-MRS npj 2D Materials and Applications (2022) 39 B. Shi et al. 11 friction under the positive gate voltage. Considering phononic
friction is a constant, the electronic friction to the relative
contribution of the two components increases with the increase
of gate voltage. Topography, friction, and adhesion measurements Published in partnership with FCT NOVA with the support of E-MRS Fabrication of the atomically thin MoS2 and AFM probe
calibration 12. Filleter, T. et al. Friction and dissipation in epitaxial graphene films. Phys. Rev. Lett. 102, 086102 (2009). The natural MoS2 bulk crystals used in this study are provided by SPI
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thick SiO2 insulation layer. The Si probes (Multi75Al-G, 3.0 N/m, Budget
Sensors) with rectangular cantilevers were used for the nanofriction and
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in silicon pn junctions. Science 313, 186–186 (2006). 29. Yan, J., Zhang, Y. B., Kim, P. & Pinczuk, A. ACKNOWLEDGEMENTS phonon coupling in graphene. Phys. Rev. Lett. 98, 166802 (200 30. Park, J. Y., Qi, Y., Ogletree, D. F., Thiel, P. A. & Salmeron, M. Influence of carrier
density on the friction properties of Silicon pn junctions. Phys. Rev. B 76, 064108
(2007). This work is supported by the National Natural Science Foundation of China (Grant
Nos. 52075093, 51775105), the Fundamental Research Funds for the Central
Universities, and the DHU Distinguished Young Professor Program. 31. Ahn, C. H. et al. Electrostatic modification of novel materials. Rev. Mod. Phys. 78,
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calibration Electric field effect tuning of electron-
phonon coupling in graphene. Phys. Rev. Lett. 98, 166802 (2007). 9. Yan, J., Zhang, Y. B., Kim, P. & Pinczuk, A. Electric field effect tuni AUTHOR CONTRIBUTIONS B.S. performed the experiments, analyzed the data, and wrote the manuscript. Y.P. and 33. Wang, Q. H., Kalantar-Zadeh, K., Kis, A., Coleman, J. N. & Strano, M. S. Electronics
and optoelectronics of two-dimensional transition metal dichalcogenides. Nat. Nanotechnol. 7, 699–712 (2012). X.G. analyzed the data and provided suggestions on the experimental design. K.Y., H.L., X.C., and K.Z. provided suggestions on the experimental design and manuscript writing. All authors discussed the results and revised the manuscript. X.C., and K.Z. provided suggestions on the experimental design and manuscript writing. All authors discussed the results and revised the manuscript. 34. Ye, J. T. et al. Superconducting dome in a gate-tuned band insulator. Science 338,
1193–1196 (2012). 35. Sarkar, D. et al. MoS2 field-effect transistor for next generation label-free bio-
sensors. ACS Nano 8, 3992–4003 (2014). COMPETING INTERESTS The authors declare no competing interests. 36. Melitz, W., Shen, J., Kummel, A. C. & Lee, S. Kelvin probe force microscopy and its
application. Surf. Sci. Rep. 66, 1–27 (2011). 37. Koren, E., Rosenwaks, Y., Allen, J. E., Hemesath, E. R. & Lauhon, L. J. Nonuniform
doping distribution along silicon nanowires measured by Kelvin probe force
microscopy and scanning photocurrent microscopy. Appl. Phys. Lett. 95, 092105
(2009). ADDITIONAL INFORMATION Supplementary information The online version contains supplementary material
available at https://doi.org/10.1038/s41699-022-00316-6. 38. Chakraborty, B. et al. Symmetry-dependent phonon renormalization in mono-
layer MoS2 transistor. Phys. Rev. B 85, 161403 (2012). Correspondence and requests for materials should be addressed to Yitian Peng. 39. Socoliuc, A., Bennewitz, R., Gnecco, E. & Meyer, E. Transition from stick-slip to
continuous sliding in atomic friction: entering a new regime of ultralow friction. Phys. Rev. Lett. 92, 134301 (2004). Reprints and permission information is available at http://www.nature.com/
reprints Reprints and permission information is available at http://www.nature.com/
reprints J. et al. Surface potential mapping of SAM-functionalized organic semi-
conductors by Kelvin probe force microscopy. Adv. Mater. 23, 502–507 (2011). 48. Wang, Y., Udyavara, S., Neurock, M. & Frisbie, C. D. Field effect modulation of
electrocatalytic hydrogen evolution at back-gated Two-Dimensional MoS2 Elec-
trodes. Nano Lett. 19, 6118–6123 (2019). © The Author(s) 2022 npj 2D Materials and Applications (2022) 39 Published in partnership with FCT NOVA with the support of E-MRS Published in partnership with FCT NOVA with the support of E-MRS
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Vers une psychopathologie du comportement antisocial chez l'enfant Vers une psychopathologie du comportement antisocial chez l'enfant Par Luis Hernan Mardones Navarro Thèse de Doctorat de Recherches en Psychanalyse et Psychopathologie Dirigée par François Richard Présentée et soutenue publiquement à Paris le 17 décembre 2016 JURY
Président du jury : Christian Hofmann, Professeur Université Paris Diderot - Paris 7 Rapporteur : Vincent Estellon, Professeur Université Paul Valéry - Montpellier III Rapporteur : Luisa Paz Rodriguez, Professeur Universidad Zaragoza Directeur de thèse : François Richard, Professeur Université Paris Diderot – Paris 7 Membre invité : Jean-Bernard Chapelier, Maitre de conférences Université de Poitiers
Résumé
Cette thèse a pour objectif d'éprouver la pertinence théorique d'un éclairage psychopathologique de la clinique du comportement antisocial chez l'enfant. Dans ce contexte, elle analyse à différents niveaux les contacts et rapports entre le psychisme de l'enfant et les discours théorico-cliniques autour de l'antisocial : psychiatrique, psychologique, sociologique, psychanalytique, phénoménologique et poétique. Dans la problématique du comportement antisocial chez l'enfant, le conflit entre la représentation du comportement de l'anti social et le récit associé à l'expérience psychique apparait comme essentiel. l'interstice de ces deux manifestations psychiques opposées, le geste est un autre type de manifestation psychique entre l'acte et le récit. Ce conflit intrapsychique s'accompagne dans la relation clinique d'une contamination du contre transfert et au niveau social, d'une riposte sociale qui est à la base de la significativité du phénomène antisocial. Notre intention est de montrer comment les modèles psychanalytiques traitant de la clinique du passage à l'acte ont laissé la question de la complexité du fonctionnement psychique de l'antisocial en périphérie. Au vu de ce constat, nous soumettons deux hypothèses: Dans la première hypothèse, nous postulerons que l'absence de significativité du comportement antisocial n'est pas simplement un court-circuit entre acte et pensée. Court-circuit dont l'incidence laisserait indemne la nature intentionnelle du comportement. La significativité antisociale est le signe psychopathologique du comportement en même temps que son intentionnalité. Abstract This research aims at underlining the theoretical relevance of the psychopathological aspect of the clinic of children's antisocial behaviour. We analyzed on different levels the contacts and relations between the child's psyche and the theoretico-clinical approaches of the antisocial: psychiatrical, psychological, phemenological, psychoanalytical and poetical. In this context, we review the importance of the conflict between the antisocial representation of the behaviour's process and the narration emerging from the experience of the antisocial psyche. With the meeting of those opposed psychic manifestations comes forward the gesture as another type of psychic manifestation between acting and thinking. In the clinical relationship, this intrapsychic conflict contamine counter-transference. At the social level, the social reaction stands at the core of the significativity of the antisocial phenomenon. Our intention is to show how the psycho-analytical models dealing with the acting out clinic overlooked the complexity of the way the antisocial's psyche works. Therefore, we propose two hypothesizes: First, the absence of significativity in the antisocial behaviour is not only a rupture between acting and thinking. A rupture which would not alter the intentionality of this behaviour. The antisocial significativity is a behavioural psychopathological marker as well as its intentionality. The antisocial gesture has to be understand as one of the morbid figures in the child's psychological development. The second hypothesis of a gerund of the gesture throws some light on the impulsive antisocial gesture. The maturity of this process comes along with the clinician progressive approximation of the antisocial phenomenon. Key-words : antisocial, psychopathie, behavior, act, significativity, gesture, antisocial gesture, gerund, game, gerund of geste, representation drive, subject. 2 A ceux dont l'amitié n'est qu'une souffrance. A mes proches auxquels ma compagnie a fait défaut. 3 Cette thèse a été possible grâce à une bourse d'excellence académique de Master recherche et Doctorat financée par La commission Nationale de Recherche en Sciences et Technologie du Gouvernement Chilien (Conicyt-Chile) et L'ambassade de la France au Chili au nom du Gouvernement Français. Je remercie le Gouvernement Chilien et le Gouvernement Français, qui ont rendu possible cette recherche à travers leur soutien financier et administratif. Je remercie le Professeur M. Jean-Bernard Chapelier d'avoir accepté de diriger ce travail de thèse, et de m'avoir soutenu avec bienveillance et confiance, dès le début jusqu'à la fin. Je remercie également le Professeur M. François Richard d'avoir été l'élan initial de cette expérience de recherche en France, ainsi que de m'accompagner pour la clôture de cette thèse. Je remercie les membres du jury d'avoir voulu juger cette thèse. Je remercie la chef de Service de pédopsychiatrie du Centre hospitalier Marc Jacquet Mme. Anne-Christine Zeegers de sa confiance sur mon travail clinico-théorique. Je remercie spécialement l'équipe professionnelle de l'Hôpital de Jour pour enfants du Centre hospitalier Marc Jacquet qui m'a permis de développer un contact plus proche des enfants au quotidien. Je remercie mes maîtres Ignacio Morlans, Rafael Parada et Edgardo Thumala. Je remercie Marco Araneda, Charles Delattre, et Alexandre Har de m'avoir accompagné amicalement, chacun à sa manière, dans cet effort. Je remercie spécialement le soutien ainsi que l'amitié de Juan Gomar.
5 REMERCIEMENTS, 5 SOMMAIRE, 6 INTRODUCTION, 8 METHODOLOGIE, 13. SOMMAIRE Partie I PROBLEMATIQUE DE L'ANTISOCIAL, 16 I. Preambule historique : etat de faits de l'antisocial, 19 II. Contact, rapport, tendance et comportement antisocial, 36
Contact antisocial, 37 Rapport antisocial, 66 Tendance antisociale, 79 Comportement antisocial Addendum : Comportement antisocial et psychopathie, 124 Psychopathie, une compréhension française de l'antisocial, 126 Description, 127 Critique, 151 Le repère psychopathique de Flavigny, 155 Partie II ANALYTIQUE PHENOMENOLOGIQUE DU GESTE, 167 I. L'enfant aux toupies,
167
Contexte clinique,
169
Description clinique, 174 Cas (anti)social, 189 Esquisse d'un diagnostic, 190 II. Acte et phrase : un tout gestuel, 213 III. Signification du tout gest
uel
, 240 La référence de Ricoeur
et la
signification de Derrida, 241 6 IV. Signification pré-psychopathologique, 277 V. Addendum : Prélude du geste, 302
Silence gestuel,
303
La grâce du geste, 307 L'idée de
suicide, enjeux esthétiques, 319 Du suicide esthétique au geste, 321 Le geste dans le théâtre :
la solution
d'Artaud,
326 Partie III SYNTHETIQUE D'UNE METAPSYCHOLOGIE DU GESTE ANTISOCIAL, 329 I.
Psycho
pathologi
e du
geste, 329 Le passage à l'acte suicidaire et son geste, 332 Au-delà de la psychose : clôturer Artaud, 341 Le geste selon Kristeva, 348 L'abject de Kristeva, 354 Geste affectif, 369 II.
Geste,
jeu et gérondif
, 381 Geste, 381 Le geste de Lacan, 382 Jeu, 398 L'enfant aux toupies avec Winnicott, 399 Gérondif, 419 Traces du gérondif dans la psychanalyse du sujet, 425 Sujet et gérondif, 441 Avec Richard, 453 Le paradigme du Nebenmensch selon Richard, 457
III. Addendum
:
Solitude antisociale,
481 CONCLUSION, 495 BIBLIOGRAPHIE, 500 7
INTRODUCTION « "Tom!" Pas de réponse [] Elle entendit un léger bruit derrière elle et se retourna juste à temps pour attraper un jeune garçon [] qu'est-ce que tu fabriquais là-dedans? – Rien
.
Comment ça, rien? Regarde
tes
mains.
Et
ta bouche! Qu'est-ce que c'est que cette
moustache? – J'en sais rien, ma tante. Mais moi, je sais! C'est de la confiture, voilà ce que c'est! »1 Les aventures de Tom Sawyer, Mark Twain (1876). « Vous me connaissez pas déjà si vous avez pas lu un livre dénommé Les aventures de Tom Sawyer, mais ça fait rien. Ce livre c'est Mr. Mark Twain qui l'a écrit, et il a dit la vérité presque toujours, avec des choses exagérées. [] La veuve Douglas a procédé à mon adoption et s'est mis dans l'idée de polir mon caractère, mais la vie à la maison était pas commode du matin au soir, vu que cette femme était si horriblement stricte sur tout. A la fin, j'en ai eu assez, j'ai mis les voiles. »2 Aventures de Huckleberry Finn, Mark Twain (1884-1885). Tout le comportement antisocial chez l'enfant se joue dans ces deux passages. Dans le premier passage, on perçoit un dialogue entre un enfant et un adulte. Il semble que l'enfant a mangé de la confiture sans la permission mais, lorsqu'il est interrogé sur le méfait, il ne peut pas rendre compte des faits, puis il se sauve. C'est tout le début du livre sur Tom Sawyer, le camarade de Huckleberry Finn. Le deuxième passage correspond à la suite du premier livre où le héros est Huckleberry, le camarade de Tom. On perçoit dans le premier passage que le récit de Tom est laconique mais la scène est bien décrite pour nous faire imaginer le type de bêtises que l'enfant fait. Ainsi, on complète l'expérience grâce à la réaction de l'adulte qui finit par interpréter ce qu'a fait l'enfant. Dans le passage de Huckleberry, c'est lui-même qui s'adresse au lecteur. Il questionne de façon ironique ce qu'a écrit Twain dans le premier bouquin. D'après nous, ces deux passages sont étroitement 1 Twain M. (1876), Les aventures de Tom Sawyer, la pléiade Gallimard, Paris, 2015, pp. 11-12. Twain M. (1884-1885), Aventures de Huckleberry Finn (Le camarade de Tom Sawyer), la pléiade Gallimard, Paris, 2015, pp. 875-876. 2 8 solidaires. Dans le premier, l'enfant ne rend pas suffisamment compte de ce qu'il a fait où ce que dit l'adulte est mis en question par le seul fait de ne « rien savoir ». Dans le deuxième, l'enfant construit un récit questionnant la parole de l'auteur du livre, c'est-à-dire revendiquant la position de l'enfant par rapports aux faits passés. En plus, Huck se montre opposé à l'idée que quelqu'un veuille « polir son caract ». Il faut préciser que dans la version originale le personnage parle dans un anglais issu d'une espèce de dialecte ou d'argot où il ne dit pas « polir mon caractère », il dit « sivilize »3, littéralement « civilize », « civiliser » en français. Il semble que pour la version française « polir le caractère » est synonyme de « civiliser ». Autrement dit, il s'agit ici d'une imposition culturelle via l'éducation. L'opposition à cette dernière peut s'exprimer par le comportement antisocial. Et si ces deux passages évoquent de la grâce, parce qu'ils sont humoristiques, ce n'est dû qu'à l'art de l'écrivain. 9 élucidation n'est qu'une opinion assujettie par l'expérience du fait que tous les enfants sont jaloux et impulsifs. La netteté de l'interprétation tombe ainsi sur une généralisation qui lui est propre, mais inexacte. Si l'on questionne l'enfant sur la motivation de son comportement il dirait, comme font la grande majorité des enfants, « je n'en sais rien ». La réponse ne rend pas compte du fait, et comparée au comportement, elle n'a rien à voir avec le contexte. On pourrait même dire que l'utilité intelligible de ces mots, c'est-à-dire sa qualité reste voilé par la grosserie du comportement. Celles-ci peuvent avoir une utilité pour l'interprétation mais, à côté de l'acte, l'utilité est marginale. Ici, ce n'est pas la force du comportement qui éclipse la force de la parole, c'est la parole elle-même que ne dit « rien ». C'est le langage rudimentaire de l'enfant qui marque la différence avec l'adulte quant à l'expression antisociale. Ni le contexte ni l'expression agis ne procurent, en effet, un sens plus ou moins établi. Si l'adulte antisocial présente un laconisme expressif quand il parle de son comportement, la parole qui s'en dégage est renvoyée au concrétisme qui est propre à l'enfant. Ce concrétisme paraît gêner l'autre qui, en questionnant l'intention de l'acte, réclame le sens voilé derrière l'évidence des faits. Mais, disons d'emblée, que suivre la trace de ce sens conduit au manque de sens ou si l'on veut, à son excès. Il semble que dans la clinique de l'enfant « difficile » la quête de sens ne manque pas au rendez-vous. Le comportement de l'enfant est optimal pour s interroger sur la nature intentionnelle du comportement antisocial. De l'acte (en passant par son passage) au comportement antisocial, selon la psychopathologie actuelle, il n'y a que le manque de langage. C'est juste mais incomplet. C'est dans cet interstice où nous situons pourtant une hypothèse forte : la manifestation psychopathologique du comportement antisocial ne se montre ni dans l'acte, ni dans la parole, mais dans le geste qui est entre les deux. Le geste antisocial présent de façon inhérente chez l'enfant surpasse, dans le comportement antisocial, l'expression évidente de la dissociation existante entre acte et phrase. Raison pour laquelle, il est impraticable aujourd'hui de continuer à postuler un paradigme tirant un sens de cette dissociation sans passer en revue la métapsychologie du sujet. L'hypothèse subsidiaire à ce raisonnement est que le comportement antisocial ne suit ni un sens rationnel linéaire (de l'acte à la phrase), ni un sens irrationnel (de la phrase à l'acte). L'antisocial est immédiatement présent dans le comportement dont la significativité est en manque. Cette dernière est presque toujours absente et son actualisation ne dépend en aucun cas de l'assignation d'un sens strictement langagier. Le problème est que la présence de significativité n'explique pas en elle-même l'absence d'une nature antisociale. L'absence de significativité du comportement antisocial, ne serait-ce qu'en raison de la 10 formation complexe des traces mnésiques et des horizons représentationnels, n'est pas simplement un court-circuit entre acte et pensée qui laisserait indemne la nature intentionnelle de ce comportement : la significativité antisociale (en négatif parce qu'absente) est déjà le signe psychopathologique du comportement et simultanément son intentionnalité. Etant présente in absentia, cette significativité est davantage repérable dans le contact avec autrui. Il faut donc s'interroger sur ce qu'elle suscite chez l'autre dont les opérations défensives vont de l'aversion paralysante jusqu'au travail sublimatoire. Le sujet antisocial semble en ce sens exprimer son symptôme par une expérience pulsionnelle irreprésentable où le rapport à autrui devient essentiel pour une possible figuration. Celle-ci est, comme la plupart des manifestations pulsionnelles, fugace et dépendante d'un pseudopode représentationnel. Il n'est pas rare alors de constater que l'appréhension dont fait preuve le sujet antisocial pour son objet, soit d'une nature sadique, car l'emprise promet de retenir et d'élargir l'évanescence de l'expérience pulsionnelle de décharge. Ce parasitisme peut être la base de l'incapacité d'intégration objectale en présupposant un processus de deuil irrésolu. Le paradoxe de décharger et de retenir simultanément chez l'antisocial détermine, dans le rapport à autrui, l'impossibilité de lier la pensée et l'acte dans un tout discursif dont le relais est repris par une instance intermédiaire qui est le geste. En priorisant le recours à l'instantanéité du geste (en réinitialisation constante et délié de son objet de récit), les instances psychiques s'atomisent chez le sujet en perdant la limite nette qui oriente le devenir psychique. En conséquence, le manque d'empathie caractéristique du moi antisocial, lié à un surmoi posé à l'extérieur, se montre mieux par l'incapacité à faire un processus de deuil : non nécessairement de l'objet perdu (dans le rapport originaire) mais de la fuite pulsionnelle persistante liée à la domination du ça. Enfin pour saisir la progression de ces idées, qui mènent vers une significativité pas strictement antisociale, une application approfondie sert de fil rouge. Nous avons confronté avec de la rigueur les approches psychanalytiques qui peuvent rendre compte du phénomène antisocial et ses déclinaisons théorico-cliniques impliquées dans une discussion métapsychologique. Ainsi, à la lumière des pathologies dites de la subjectivation, nous avons pu confronter notre démarche avec la psychanalyse actuelle et attaquer les positions qui rendaient compte des enjeux autant métapsychologiques que des relations intersubjectives. 12 METHODOLOGIE
Cette thèse s'inscrit dans une démarche d'articulation clinique et théorique. Elle trouve sa base dans l'expérience clinique qui annonce le phénomène psychopathologique, mais qui tend à le subjuguer dans la contingence professionnelle contextuelle. Par contingence professionnelle, nous entendons les dispositifs cliniques où se construit le contact avec le phénomène antisocial. Le dispositif clinique et institutionnel en fonction de sa temporalité (urgence, crise, chronicité) figure la compréhension du phénomène psychopathologique et son traitement. Pour chaque illustration clinique que j'ai choisi de développer, les lieux et les fonctions seront précisés : Au Chili, « Previene-CONACE » est un dispositif communautaire de prévention de la consommation de stupéfiants dans lequel j'ai développé des interventions psycho-éducatives pour des populations à risque enfants et adolescents auprès des mairies, des écoles primaires et secondaires. Responsable du programme d'accueil et de traitement pour toxicomanes, « Service de santé aire nord » où j'ai occupé le poste de psychologue clinicien. Service d'addictologie de « l'Institut Psychiatrique Dr. Horwitz Barak » où j'ai eu le rôle de psychologue clinicien hospitalier. Psychodiagnostic et psychothérapie, en pratique libérale. En France, Centre de Soins, d'Accompagnement et de Prévention en Addictologie, Emergence, « Institut Mutualiste Montsouris », en tant que psychologue clinicien. Hôpital de jour pour enfants, « Centre Hospitalier Marc Jacquet » où j'exerce actuellement en tant que psychologue clinicien. La logique méthodologique : 13
Le champ de travail de cette thèse « l'antisocial chez l'enfant », s'organise selon trois axes méthodologiques : clinico-descriptif, théorico-conceptuel et poético-esthétique. Chaque axe sera questionné selon plusieurs dimensions de réflexion que nous préciserons au fur et à mesure. L'axe clinico-descriptif s'appuie sur une illustration d'un cas clinique détaillé auquel viendront s'ajouter des vignettes cliniques, selon la pertinence du sujet déroulant. Le choix de cette présentation et de cette mise en forme est dû à plusieurs raisons. D'abord je souhaite mettre en parallèle les expressions cliniques concernant l'antisocial, à celles qui sont réduites à un syndrome de type psychopathique. Puis les illustrations cliniques seront complétées d'une lecture transnosographique du phénomène antisocial. Le matériel clinique est tiré de plusieurs expériences thérapeutiques, son origine sera signalée pour avertir du contexte où se déroule la manifestation antisociale. La sphère descriptive permettra d'observer le comportement du phénomène antisocial à plusieurs niveaux : une manifestation comportementale à l'âge adulte et pendant l'enfance, et selon plusieurs niveaux de lectures psychanalytique, psychiatrique, psychopathologique, psychologique. Nous montrerons également les mutations du phénomène antisocial et ses contextes. Cette mutation non exclusive à l'antisocial soulève d'autres questions méthodologiques, celle de l'âge de l'émergence des troubles et celle de la comorbidité. L'axe théorico-conceptuel abordera l'examen critique du phénomène antisocial. La revue des différents courants théoriques en psychanalyse et leur révision conceptuelle permettront d'élargir la discussion strictement clinique. Cette analyse sera confrontée avec d'autres disciplines proches de la psychanalyse afin d'éclaircir le phénomène clinique et d'éviter les solutions de type cosmétique, qui tendent à l'obscurcir. La philosophie contribuera à l'extension et à la précision de la nature esthétique du phénomène antisocial. L'axe poético-esthétique viendra traiter des manifestations poétiques du comportement antisocial. Différentes sources seront consultées dans lesquelles le phénomène antisocial s'exprime avec netteté. La littérature aura une place importante pour ce repérage. A partir d'extraits de la littérature : romans, poésies, correspondances, scénarios de films, mon intention sera de mettre en exergue les manifestations antisociales. Ces trois dimensions connaîtront un traitement analytique et synthétique. A savoir, l'analyse concrète de leurs expressions textuelles, c'est-à-dire la description du propos quant à sa forme, origine contextuelle et les implications hors du champ de leurs disciplines. De cette façon, les courants de pensée seraient considérés moins comme des vérités établies que comme 14 des outils conceptuels sujets à une critique en progression. Partie I PROBLEMATIQUE DE L'ANTISOCIAL
Il est difficile, dans la clinique, de prendre contact avec l'antisocial. Un fait connu par les cliniciens soutient cette affirmation : les psychopathes ne consultent pas. Un autre fait suit celui-ci : les expressions antisociales du psychopathe peuvent être repérées par le biais d'un autre trouble. Cette situation met le clinicien en contact avec l'antisocial du psychopathe ou l'antisocial d'un patient quel qu'il soit. Mais cela n'est pour autant pas un contact direct avec la nature antisociale pure. Prenons un exemple pour illustrer ces propos. L'un des critères d'exclusion pour le traitement de désintoxication des patients toxicomanes est la psychopathie. Le patient présentant une personnalité dite psychopathique ne peut pas se faire hospitaliser pour une cure de désintoxication. Il n'a l'accès qu'à un accueil de type ambulatoire. Les chances de réussir à une désintoxication se voient amoindries et le cycle de consommation devient perpétuel. Ce cycle compulsif restreint la marge de manoeuvre comportementale du patient. Il ne vit que pour consommer de la drogue. Cette compulsion transforme la personnalité du patient en la réduisant à un circuit hermétique dans lequel le centre d'intérêt est la préservation de la satisfaction liée à la drogue. Une réaction violente suit cette satisfaction si elle se voit interrompue. Cette réaction est presque normale dans tout processus de désintoxication. On peut même la préconiser pour lui faire face. Le propos de réagir contre la désintoxication (en passant à l'acte) vaut contre toute intention à tenir une abstention pour issue. En effet, le patient lutte contre le syndrome de privation, c'est-à-dire il vit comme nécessaire la résistance même si elle implique le dommage de lui-même et de son entourage. C'est dans ce d que se manifeste initialement l'antisocial. En effet, passer à l'acte violemment en agressant, en mentant, en volant figure l'attaque antisociale à l'autre. Ce comportement antisocial est un couteau à double tranchant. D'un côté, la quête effrénée de drogue active violemment le recours à l'acte, c'est-à-dire qu'elle est entièrement justifiée par le sujet agissant. D'un autre côté, celle-ci prive le sujet du soutien nécessaire pour se faire guérir. La souffrance liée au corrélat somatique est éclipsée par le comportement antisocial. Ce corrélat somatique occulte à son tour la vraie intentionnalité du comportement antisocial. Le clinicien qui travaille en addictologie connaît de surcroît les 16 vicissitudes liées à la consommation de drogue. Il priorise la suppression de symptômes somatiques en même temps qu'il motive le patient (et son entourage) au changement, puis il analyse les facteurs de risques d'une possible récidive. Généralement, dans cette analyse il questionne la nature intentionnelle du comportement antisocial. La plupart du temps, celle-ci corrobore sa liaison à la quête de drogue. Le mot-clé pour cette affaire est la comorbidité ou la cooccurrence de deux ou plusieurs pathologies psychiatriques. Ce qui importe est la suppression de la symptomatologie addictive et cela est sans aucun doute juste. Ainsi, ce qui pose problème pour le traitement d'une addiction, c'est le comportement antisocial. Mieux, ce dernier pose problème à tout type de traitement (principalement thérapeutique). Ce comportement éloigne le patient d'une quête de secours franche, condition essentielle pour recevoir un accueil bienveillant. Sous cette logique, l'entourage du patient s'étend de sa famille vers l'institution. Tout le monde est concerné par la dépendance du patient à la drogue et ultérieurement à l'institution. Cette dernière rejette le comportement antisocial du patient au même titre que le fait la famille. Avec cet exemple nous voulions d'abord illustrer la nuisibilité du comportement antisocial. Puis, montrer comment celui-ci est un critère d'exclusion, même si une pathologie grave comme la toxicomanie l'amène à l'examen. Mais, ce que nous voulions préciser, c'est le rejet social qu'il évoque. Ce rejet social est le seul point en commun avec le comportement antisocial chez l'enfant. En fait l'enfant, avec des comportements antisociaux, est presque sans doute considéré malade et son exclusion sociale devient la porte d'entrée dans l'institution psychiatrique. Chez l'enfant – en assumant qu'il ne consomme pas de stupéfiants par imitation de l entourage – tout ce qui était considéré comme critère d'exclusion chez l'adulte, sert de motif de traitement du comportement antisocial. Ce dernier est considéré à part entière comme un signe de morbidité chez l'enfant. Il semble qu'être en contact avec ce type d'enfant soit moins nuisible que de l'être avec l'adulte. Encore, il semble que si ce contact n'est pas possible, un rapport ultérieur est impensable. Est-ce que ce passage du contact au rapport est toujours univoque? Est-il possible d'établir un rapport avec l'antisocial adulte? Ou est-il nécessaire de comprendre l'évolution entre contact et rapport, autrement? Pour répondre à ces questions, il faut observer comme se comporte le comportement antisocial lorsqu'il entre en contact avec autrui. PREAMBULE HISTORIQUE : ETAT DE FAITS DE L'ANTISOCIAL
L'antisocial chez Freud se montre de manière flagrante, au contraire de ce que l'on peut fréquemment supposer, non spécialement par ses formulations théoriques au respect, mais plutôt par son versant technique. En fait, Freud prononce en 1906 une conférence Diagnostic de l'état des faits et psychanalyse4 en faisant une comparaison entre le psychopathologique et le criminel. Ce texte n'offre pas de grandes difficultés au chercheur lorsqu'il veut déchiffrer les propos criminologiques qui en découlent. De plus, la manière didactique à laquelle Freud soumet les topiques est claire et illustrative. Même, s'il s'agit de travailler les notions plus problématiques des « idées incidentes » liées à « l'association » libre – déjà respectivement différenciées par Freud entre Einfall et Assoziation –, ce texte n'offre pas une résistance majeure au lecteur. Il suffit, pour surmonter toute complication possible de rapprocher ces notions à l'Interprétation de rêves, Sur psychopathologie de la vie quotidienne et à La technique psychanalytique pour savoir que Freud voulait mettre en exergue l'intentionnalité inconsciente qu'il y a derrière les associations sauvages5. Pourtant, au moment de le travailler en filigrane, d'emblée, le titre en langue originale « Tatbestandsdiagnostik und psychoanalyse » mérite une petite décomposition pour souligner la problématique de son propos. Tatbestandsdiagnostik, mot composé de Tatbestands (état des faits)/ diagnostik (diagnostic) nous offre un sens littéral : Diagnostic de l'état des faits. Mais si l'on y regarde de plus près, on note que le diagnostic est proche du judiciaire, certes on peut faire un diagnostic de quelques états de fait, sans nul doute. Pourtant, et voilà la logique du titre, la présence la psychanalyse nous montre comment la transposition du diagnostic, généralement proche du clinicien, passe du côté du judiciaire. Les traductions précédentes montrent des enjeux particuliers, la première de 1933 La psychanalyse et l'établissement des faits en matière judiciaire par une méthode diagnostique n'admettait pas de malentendus de traduction, elle se faisait directement à l'interprétation explicite, tout en laissant de côté la problématique que présente le titre. La seconde de 4 Freud, S. (1906) « Diagnostic de
l'
état des fait
s
et psychanalyse » in OEuvres complète
s
. Psychanalyse. Tom
e
VIII
(
1906
-
1908
),
Puf
,
Paris, 2019.
Cette conférence apparaît écrite
in "Archiv für Kriminal-Antropologie und Kriminalistik" périodique dirig
é
par Hans Gross, professeur de criminologie à Prague
(p.14). L'établissement des faits par voie diagnostique et la psychanalyse semble être plus fidèle au titre allemand, mais elle se montre à son tour proche de la traduction anglaise de 1959 Psycoanalysis and the Establishment of the Facts in Legal Proceedings. Les deux premières parlent d'établissement des faits via le diagnostic, toutes les deux font une adjectivation du diagnostic pour établir ou fixer les faits via un diagnostic. Alors que la version anglaise fait l'économie du mot diagnostic, mais elle ajoute aussi le mot établissement, Establishment. Certes les trois traductions cherchent à rendre justice au titre freudien, mais tout en ajoutant le mot établissement comme si, par l'exercice psychanalytique, on pourrait déterminer ou établir les faits de manière judiciaire. La version actuelle avec laquelle nous travaillons ce texte est Diagnostic de l'état des faits et psychanalyse, elle semble être la plus fidèle au titre original car elle combine littéralité et sens, sans besoin d'ajouter le mot établissement. « Le diagnostic des faits » fait trait d'union avec ce qui est écarté par le « et ». Faire le diagnostic implique alors dans un second degré, la participation de la psychanalyse dans l'établissement des faits. Cependant, assumer un certain établissement, signifie grosso modo penser à une psychanalyse appliquée au champ judiciaire. Contrairement, il semble que l'intention de Freud était, depuis le début du texte, de garder à distance la psychanalyse du judiciaire, même si les deux disciplines partagent une méthode, voire une technique – à l'époque, l'application des « expériences d'association ». Cette appréhension ne quittera jamais Freud. Il était aussi délicat à ce propos, qu'en 1931 dans L'expertise de la Faculté au procès Halsmann6, Freud n'hésite guère, même si la confirmation du complexe d'OEdipe étai en jeu, d'en dénoncer l'usage inapproprié de la psychanalyse en matière judiciaire. Philipp Halsmann avait été jugé en 1929 pour le meurtre de son père. Erich Fromm écrit un article en questionnant l'usage du complexe d'OEdipe comme élément à charge contre Halsmann. Freud commence le texte en citant « Le neveu de Rameau » de Diderot, à propos d'un petit sauvage qui tuerait son père et puis coucherait avec sa mère. Freud met ici l'accent, à juste titre, sur l'imbécillité du sauvage tout en montrant le penchant à l'agression que garde le complexe d'OEdipe, mais surtout sur l'imbécillité qui régit le contrôle de la volonté agissante – dont les facultés mentales sont grossièrement perturbées. 6 Freud S. (1930) « L'expertise de la Faculté au procès Halsmann » in OEuvres complètes Psychanalyse tome XIX, Puf, Paris, 1995, pp.39-43. Freud justifie son désaccord avec le jugement « la mention du complexe d'OEdipe induit en erreur. »7 Puis il rappelle une anecdote, un vol avec effraction – jeu de mots entre Einbruch (effraction) et Ehebruch (infraction au mariage)8 – dont un accusé réclame d'être puni pour adultère, car il porte aussi l'instrument sur lui. Dans cette anecdote, bien proche, elle, d'un « trait d'esprit », Freud conteste encore l'usage abusé du complexe d'OEdipe. Mais c'est sur ce jeu de mots, sensé d'avoir une significativité, que l'évidence intentionnelle tombe sur l'absurde, voire la surinterprétation. Avant de reprendre l'expertise en traitement, Freud fait une dernière référence : Les frères Karamazov de Dostoïevski. Le traitement qu'il en fait ne peut être classé que brillant. Freud montre brièvement l'action du complexe d'OEdipe dans le déroulement de la trame, mais, dans un même coup, il remarque l'ambiguïté de son usage, en citant « la psychologie est une arme à double tranchant.»9 Ainsi, et après ces trois références, Freud prononce son jugement, qui mérite une citation in extenso : « Dans cette même expertise, on se heurte à une contradiction qui est loin d'être sans significativité. L'influence possible de l'ébranlement de l'humeur sur le trouble de la mémoire portant sur les impressions avant et pendant la période critique est réduite à l'extrême, à tort selon mon jugement ; les hypothèses d'un état exceptionnel ou d'une affection animique sont résolument rejetées, mais l'explication par un "refoulement" intervenu chez Philipp Halsmann après l'acte est complaisamment admise. Je dois dire qu'un tel refoulement tombé d'un sans nuages chez un adulte n'offrant aucun signe de névrose grave – refoulement d'une action, qui serait à coup sûr d'une plus grande significativité que tous les détails contestables d'éloignement et de chronologie et qui se produit dans un état normal ou simplement modifié par la fatigue du corps –, voilà qui serait certes une rareté de premier ordre. »10 Cette « significativité », si souhaitée pour justifier ce fait criminel, n'est ici présente que in absentia. Et elle, si remarquée par Freud, fait partie du chaînon, d'une chaîne d'événements liés par une cause sans signification efficiente. Ainsi, ce que réclame Freud n'est pas seulement l'usage du complexe d'OEdipe, mais une significativité inappropriée. Cette significativité n'est pas autre que la Bedeutung.11
7 Ibid., p.41. Ibid., p.42, note a de traducteurs. 9 Ibid., p.42 10 Ibid., pp.42-43. Nous soulignons. Dans la traduction de 1985 ce que nous soulignons, était traduit pour (sans) importance Bedeutungslosen et signification Bedeutsamer. Freud S. « L'expertise de la Faculté au procès Halsmann » in Résultats, idées et problèmes, II, Puf, Paris, p.189. 11 Dans Traduire Freud à ce propos « pour Bedeutung : le français hésite à chaque fois entre "signification" et "importance", une question que ne se pose pas, en revanche, le lecteur anglais pour qui le mot signifiance recouvre assez exactement la totalité de Bedeutung. Bourguignon A., Cotet P., Laplanche J., Robert F., Traduire Freud, Paris, Puf, 1989, p.52. Les auteurs vont diviser l'utilité du mot, Bedeutung comme « importance » et/ou Bedeutung comme « signification » à ce qu'ils déclinent en « significativité » pour mieux saisir la différence de son emploi. Ainsi, « comme substantif dérivé en –ung, ce mot peut désigner soit le résultat achevé d'un processus (signifier La Bedeutung significativité, auparavant traduite comme l'importance, est la vraie « arme à double tranchant ». La contradiction que Freud remarque, est une significativité en erreur, c'est-à-dire l'accentuation d'un fait qui n'a pas d'importance capitale dans le déroulement de l'action. En d'autres termes, elle y participe et a même un pouvoir d'explication, mais ce n'est pas la cause juste. Métaphoriquement parlant, ce serait comme un mot mal accentué, par exemple « chanter », à l'oral il souffre la suppression de la lettre « r », puis il gagne l'accent dans la voyelle « é », mais « chantér », incorrectement écrit, n'a aucun sens efficient, puisque la transformation s'est déjà fait à l'oral, en consé l'ajout devient une contamination de genres. Ainsi, l'écriture correcte du mot « chanter » est, sans faire mention du contexte, subjuguée à l'interprétation de lecture dont l'accent participe, pour ainsi dire de manière virtuelle. De sorte que cet usage du complexe d'OEdipe explique partiellement l'action dont la contradiction saute aux yeux en tant que cible ratée. A contrario pour élucider le fait, cette signification le recouvre d'une explication si spéculative que l'acte brut devient une subtilité trompeuse et impropre. Cette significativité ratée veut dire justement une autre chose : ce qui était significatif chez l'accusé (période critique de l'acte) a perdu sa valeur à force de l'interpréter comme un refoulement oedipien. Suite à cette parenthèse, nous pouvons aborder le texte en question, car, au-delà du criminel, les deux textes sont étroitement liés. Ce contenu de représentation correspond à ce que Jung appelle « complexe » – que Freud avait déjà esquissé dans Études sur l'hystérie (1894-1895) pour désigner des souvenirs liés par association. Tout de suite, Freud lie ces associations à ses travaux de 1901 (Sur la psychopathologie de la vie quotidienne) pour argumenter les motivations inconscientes, « sens caché » dit-il, des méprises quelles qu'elles soient. C'est là justement que la distinction entre « idées incidentes » et « expériences d'association » est cruciale, car les premières sont liées à une occurrence, on dirait fortuite, sans qu'il y ait une médiation de par la volonté du sujet dans l'exercice associatif ; tandis que dans les secondes, le sujet reste attentif, voire collaborateur au processus associatif qui lui arrive. Les premières sont, on dirait, plus proches des opérations manquées, alors que les secondes sont voisines de la libre association. C'est la critique qui fait la différence entre les deux, sa présence déterminera la censure de l'idée incidente, et des pensées non voulues, tandis que son absence déterminera un discourir continuel de ces pensées et ainsi passer à une association « libre ». Cette dernière faculté n'est pas toujours à portée de main du sujet, la plupart du temps elle souffre de fortes résistances. Pour rendre encore plus claire l'importance de cette distinction dans le texte, il faut citer un passage de l'interprétation de rêves où Freud fait une comparaison entre l'homme qui se donne à la réflexion et celui qui fait l'auto-observation de ses processus psychiques : « Dans la réflexion, une action psychique entre davantage en jeu que dans l'autoobservation la plus attentive, comme le prouvent d'ailleurs la mine tendue le front plissé de celui qui réfléchit, à l'opposé de la mimique tranquille de l'auto-observateur. Dans les deux cas, il y a forcément une concentration de l'attention, mais celui qui réfléchit exerce en outre une critique, par suite de quoi il rejette, après qu'il les a perçues, une partie des idées incidentes montant en lui, coupe court à d'autres, de sorte qu'il ne suit pas les chemins de pensée qu'elles ouvriraient, tandis qu'envers d'autres pensées encore il sait s'y prendre pour qu'elles ne deviennent absolument pas conscientes, donc qu'elles soient réprimées avant d'être perçues. L'auto-observateur, par contre, n'a que 23 la peine de réprimer la critique ; s'il y réussit, une multitude d'idées incidentes lui viennent à la conscience qui, sinon, seraient restées insaisissables. » 13 Freud fait l'analogie de cet état psychique avec celui du préambule à l'endormissement. On serait tenté de dire une véritable hallucination hypnagogique, cependant dans ce processus décrit plus haut, il y a une instance critique qui régit – facultativement – la réception des idées incidentes. De plus, celles-ci semblent flâner sur l'espace de conscience, mais leur présence, bien qu'elle soit proche du hasard, n'est que requise par une instance observatrice. Une déformation extrême de ces idées, serait de penser à une agglomération des idées dans la conscience du sujet. Telles quelles, ce seraient des pensées en amoncellement dues à un changement de l'humeur et sa conséquente tachypsychie et hyper-condensation des pensées en fuite – dont leur sens est plus quantitatif (vitesse et accélération) que qualitatif (association et symbolique). Pourtant ce que Freud pointe là est tout à fait d'une autre nature : le sujet a un certain gain dans l'observation du flux idéatif. On dirait un exquis équilibre entre contrôle et relaxation où l'intentionnalité circule dans le courant idéo-affectif. Ainsi, ce que Freud montre, via l'exemple mécaniciste de stimuli-réaction de l'école de Zurich, n'est pour autant pas un processus mécanique de pensée associative, il est l'émulsion juste entre production et résultat. Ce qui est important dans le texte que nous traitons, c'est que la peine à réprimer la critique stimule un processus de pensée spontanée et libre. C'est ici que Freud met l'accent pour différencier le criminel du névrosé. d'abord Freud dit que le secret est pour l'hystérique quelque chose d'involontaire, qu'il méconnaît, tandis que le criminel est conscient et ne restreint sa connaissance qu'à lui-même, il le cache à autrui. Freud montre que cette différence est fondamentale lorsqu'on interroge ou demande au sujet d'associer les mots-stimuli. Le criminel ne fait pas d'expérience spontanée, on dirait qu'il est préparé à répéter identiquement ses associations tout en occultant la réaction au complexe – ce qui le ferait réagir ou ce qui déclencherait une libre association. Bref, il est actif dans sa critique, voire il la fait travailler en sa faveur et en même temps contre l'autre. Alors que le névrosé a une attitude de travail pour le clinicien, c'est-à-dire en faveur de la guérison, souligne Freud dans le texte. 24 aura une conscience de culpabilité agissante, une « mauvaise conscience » dit Freud ; on peut par conséquent supposer que le criminel en est dépourvu. Avant d'analyser la différence qualitative qui existe entre l'un et l'autre, nous voulons mettre en relief une différence quantitative. En fait, la quantité de temps qu'il y a entre le motstimulus et les idées incidentes – quasiment toujours plurielles sous la plume de Freud, car ces idées appartiennent à un réseau associatif mixte, c'est-à-dire subjuguées aux processus de condensation et déplacement – est déterminante pour comprendre la distinction entre criminel et malade. En effet, Freud montre comment le malade « s'arrête », « hésite » et « fait des pauses » dans la reproduction des idées incidentes. Il catalogue ce phénomène comme une « résistance », un signe lié au « complexe » : « Elle [la résistance] est même pour nous l'indice le plus important d'une telle signification, tout comme pour vous [le public] l'allongement analogue du temps de réaction. »14 Ce passage ne parle pas seulement de la résistance au sens clinique, il montre aussi la transposition de la clinique au judiciaire. Ce temps de réaction dans le domaine judiciaire reste sans qualification psychopathologique, tandis que dans la psychanalyse il peut être qualifié de résistance. Ici on voit très clairement comment Freud tranche les différents espaces, mais ce qui en résulte de plus intéressant, c'est d'analyser la manière dont il le fait. Par ailleurs, Paul-Laurent Assoun a un avis semblable. Pourtant l'auteur examine le texte en parlant de synonyme là où nous avons noté une différence – surtout à propos du titre. Cela va avoir des conséquences d'appréciation car Assoun privilégierait les sujets classiques suscités par le texte : « Ce texte est l'occasion de r la notion de COMPLEXE [] cela permet de mettre en évidence la règle de LIBRE ASSOCIATION. Enfin, la notion de SENTIMENT DE CULPABILITÉ, en sa dimension inconsciente, se trouve restituée par une mise en regard de ses formes névrotique et criminelle. »15 Certes le texte traite toutes ces notions-là, même si elles sont mises en tension entre une discipline et l'autre. Toutefois, et bien que la systématisation d'Assoun les mette clairement en exergue, elle ne cerne plus suffisamment un sujet en filigrane traité par Freud. L'accent mis par Freud, que nous soulignons plus haut, n'est pas anodin du tout, ni sommairement rhétorique non plus. psychopathologique entre le criminel et le malade. En effet, la résistance « indice plus importante d'une signification » regroupe, dans son sein, le zeigen et la Bedeutung, c'est-à-dire, l'indice de la significativité. Sans ces notions, la résistance montrée, soit par le criminel ou par le névrosé, n'aurait pas de ressemblance, ces derniers seraient alors, pour ainsi dire, condamnés à marcher en parallèle, tout en ayant une nature psychologique bien similaire. Cette similitude est loin d'être sans implications, puisqu'elle fait partager la significativité qui a un par rapport de l'une à l'autre. Autrement dit, la significativité qu'a le malade passe pour ainsi dire du côté du criminel, et ce dernier à son tour partage la sienne avec le premier. D'où la technique qui montre la différence qui fait liaison entre judiciaire et psychanalyse. On pourrait alors dire que la matière antisociale prend la forme de celui qui l'exprime. De plus cette expression, pure dans ses origines en tant qu'intention, subit une deuxième mutation, celle de l'interprétation. Un exemple simple, sans rentrer dans des spécifications majeures, est de penser à ce qu'on entend par Acting-out et passage à l'acte ; le premier trouve sa significativité dans l'interprétation, mais surtout grâce au contexte du cadre psychothérapique, alors que le deuxième, aussi dépendant de l'interprétation, est livré à l'intempérie dont la significativité est partagée avec la réalité matérielle. Le premier a la symbolisation psychanalytique en sa faveur, le deuxième est en dépourvu. Ce dernier est tout à fait interprétable, mais exposé à la sauvagerie d'interprétations. L'Acting-out, même s'il vise la fuite, garde toujours un domicile connu, alors que le à l'acte fait sa demeure dans les lieux de transit. L'orphelinat du dernier est le sceau de l'antisocial. Nous avons dit que ce texte était tiré d'une conférence faite pour Freud aux étudiants de Droit pénal du Pr. Löffler, donc un public profane à la doctrine psychanalytique. Cela nous permet de faire un petit détour explicatif. Le sujet à traiter était très pertinent aux deux disciplines car il s'agissait de la mise en pratique d'un protocole psychologique pour faire les interrogations aux inculpés. De plus, Freud cite Hans Gross réputé criminologue et directeur du périodique où était publiée cette conférence. Ce dernier était aussi père d'Otto Gross, médecin psychanalyste, traité par Bleuler puis Jung à l'hôpital Burghölzli à Zurich. 26 Jungienne. En fait, dans l'avant-propos du livre qui groupe ces recherches, daté de juillet 1906, Jung reconnaît à son tour l'influence de Freud « un coup d'oeil même rapide sur les pages de ce travail montre ce que je dois aux géniales conceptions de Freud. »17 Tout à fait juste car Jung fait, bien que psychiatrique dans ces descriptions, une interprétation psychanalytique des processus inconscients agissant dans les expériences d'association. Il passe en revue une série très large de positions psychopathologiques et aborde également la littérature dominante de l'époque à propos de la démence précoce : « détérioration aperceptive (Weygandt) ; dissociation, abaissement du niveau mental (Janet, Masselon) ; désintégration de la conscience (Gross) ; désintégration de la personnalité (Neisser et al.) [] tendance à fixation (Masselon, Neisser) et Neisser en déduit l'appauvrissement affectif »18. Jung n'hésite guère à remarquer les travaux de Freud, tout en les associant à ceux de Gross : « Freud et Gross découvrent le fait important de l'existence de séries de représentations clivées. »19 Jung en parle, à propos d'un cas très détaillé d'une patiente atteinte de démence précoce : « des complexes opposés refoulés, d'un côté du complexe de préjudice, de l'autre des restes de correction normale. L'entrée dans la conscience de fragments de ces séries devenues autonomes se réalise essentiellement sous forme d'hallucinations, à la manière décrite par Gross et à partir des racines psychiques supposées par Freud. »20 Bien que Jung remarque l'importance des travaux de Freud « Freud a dit tout l'essentiel dans ses travaux sur l'hystérie, la névrose obsessionnelle et le rêve »21, l'auteur s'en détache partiellement en ant la notion de « complexe à tonalité affective. »22 Jung précise : « Le fondement essentiel de notre personnalité est l'affectivité. Pensée et action ne sont pour ainsi dire que des symptômes de l'affectivité. »23 De cette citation on peut déduire rapidement que Jung considère l'affect comme quelque chose d'inconscient, une sorte de propulseur primordial de la vie psychique dont l'action et la pensée en sont les représentants. Il est clair que l'on est en présence d'une séquelle expérimentale de la triade, structure primordiale des 17 Jung C.J. (1907), Psychogénèse des maladies mentales, Albin Michel, Paris, 2001, p.13. Ibid., p.52. Cf. Jones E. (1955) La vie et l'oeuvre de Sigmund Freud. II Les années de maturité 1901-1919, Puf, Paris, 2006, Selon Jones, Otto Gross en 1904 a écrit « Zur differentialdiagnostic negativischer Phänomene » où il compare la dissociation des idées, décrites par Freud, et la dissociation dans l'activité consciente chez les démences précoces. A cet article a suivi un livre en 1907: « Das Freudische Ideogenitätsmoment und seine Bedeutung im manisch-depressiven Irresein Kraepelins » (p.32). 18 19 Jung C.J. (1907), Psychogénèse des maladies mentales, op. cit., p.52. Ibid., pp.180-181. 21 Ibid., p.53. 22 Ibid., p.53. Ici on voit comment les conceptions jungiennes, même s'il dit se différencier légèrement, sa logique reste très freudienne, surtout si l'on songe au Projet d'une psychologie de 1895 de Freud. 23 Ibid., p.53. 20 27 comportementalistes – cognition, comportement, affect – héritiers de l'école de Wundt surtout, pourtant Jung dépasse ce courant. Parce qu'il argumente les associations par une intentionnalité inconsciente inspirée principalement de Freud – et dans une considérable mesure de Janet à propos d'un appauvrissement de l'attention. De cette manière, Jung justifie les rythmes, latences, intensités des réponses des associations, les erreurs – un fragment serait tout à fait dédié aux lapsus calamis, les « lapsus freudiens » comme dit Jung. Ce dernier précise les expériences d'association : « Le sujet a reçu pour consigne de porter son attention sur l'expérience ; si son attention faiblit, c'est-à-dire si elle se détourne de la signification du mot inducteur sans qu'aucune cause externe puisse être incriminée, il doit nécessairement exister une cause interne de cette distraction : cette cause, nous la trouvons le plus uvent dans la réaction précédente ou encore dans la même réaction. Une idée à forte charge affective est apparue, un complexe, qui en raison de sa forte tonalité affective, acquiert une grande netteté dans la conscience ou, en cas de refoulement, fait passer une inhibition dans la conscience, supprimant ou diminuant par là pour un court moment l'efficacité de la représentation donnée en orientation (attention portée au mot inducteur). L'exactitude de cette hypothèse se laisse le plus souvent démontrer sans peine par l'analyse. »24 Ce passage offre une confirmation de ce que nous avons signalé plus haut, par rapport à la mécanique du mot-stimulus et réaction que Freud décrit dans sa conférence – description de « l'application scientifique » de ses propres idées. De plus ce passage montre le coeur de la technique junguienne du « test d'association », c'est-à-dire un outil – physico-conceptuel = réaction-galvanomètre – qui, dans son usage, surtout au début, n'était guère limité à la clinique. Il faut aussi ajouter que cette citation est accompagnée d'une note qui renvoie à deux travaux précédents de Jung. C'est dans l'un d'eux que nous remarquons le titre « Die psychologische Diagnose des Tatbestandes » 1906 traduit comme « Le diagnostic psychologique des faits » ; ce titre est assez ressemblant à celui de Freud de 1906, ce n'est pas un hasard parce que c'est à propos de ce texte que Freud fait ici la première référence à Jung25. L'autre texte de Jung « Diagnostische Assoziationsstudien »26 regroupe ses travaux initiaux sur les expériences 24 Ibid., p.70. Cf. L'introduction de Freud S., Jung C., Correspondence 1906-1914, op. cit., p.12. Selon Bair, en interprétant Jung : « un criminel impliqué dans une affaire sait comment les choses se sont déroulées et en viendra « inconsciemment » à se trahir [] Jung formulait cependant une « objection solide à l'utilisation médico-légale de cette méthode » ». Bair D., Jung, op. cit., p.148. On songe ici aux mêmes précautions prises par Freud à propos du texte en traitement, mais surtout à L'expertise de la faculté au procès Halsmann, cité en haut. Pourtant, d'après Ellenberger, à propos d'un vol d'argent dans un hôpital, Jung soumit les soupçonnés au test d'association. Ellenberger H. F., A la découverte de l'inconscient, Simep, Villeursbanne, 1974, p.573. De plus, Hugo Munsterberg, professeur de psychologie à Harvard, utiliserait, la permission de Jung
, les
expériences d
'
association
en
milieu judiciaire
. Bair D., Jung
,
op
.
cit., p.151. 26 Jung C.J. (1911-1913), Psychogénèse des maladies mentales, Albin Michel, Paris, 2001, p.361. 25
28 d'association. Mais ce sujet avait déjà des extensions précédentes sur d'autres domaines. Le 21 d'octobre 1905, Jung donne son premier cours sur « La signification psychopathologique des expériences d'association » lequel sera, publié une année après avec le même titre, dans le périodique de Hans Gross27 Archiv für Kriminalanthropologie und Kriminalistik (Leipzig) volume 2228. Freud y écrira comme nous l'avons déjà dit, dans le volume 26. Partager ce périodique ne fait pas seulement que rapprocher les auteurs quant aux sujets scientifiques (psychopathologie et criminologie), il les lie surtout par la clinique. 27 Considéré comme
le père
de
la
criminologie et de la psychologie judiciaire. Bair D., Jung, op. cit., p.147. Voir note 47 Ibid., p.1028 (« Die psychopathologische Bedeutung des Assoziationsexperimentes »), nous soulignons. 29 Freud S. (1906) « Diagnostic de l'état des faits et psychanalyse » in OEuvres complètes. Psychanalyse. Tome VIII, op. cit., pp. 23-24. 30 Ibid., p.24. Nous soulignons. 28 29
« Établir
par des expérience
s »
d'association
« si
la résistance consciente se tra
hit » rompt bel et bien les ponts entre
la
clinique et
le judiciaire. « Établir »
reste du côté
du
judiciaire
: la
résistance est
à
la frontière
de l'inconscient et du conscient, mais aussi entre les deux disciplines ; elle
est établi
e
dans
les
état
s de faits alors que dans la clinique elle est diagnostiquée. Le problème ici c'est que cette « résistance consciente », lors de se trahir, passe du côté inconscient – ce qui n'est pas la même chose que de passer du côté de la clinique –, ce qui revient au même de dire, transiter impunément d'un côté à l'autre. Elle peut parfaitement être interprétée comme une conscience de culpabilité des deux côtés. La clinique interprète ce fait comme quelque chose de névrotique, alors que le judiciaire comme un fait criminel. La significativité de ce phénomène se trouve à la base de la demande théorique faite par la discipline judiciaire envers la psychanalyse. Depuis l'année 1880, souligne Ellenberger, Hans Gross « avait noté des lapsus linguae significatifs et d'autres manifestations analogues [] Gross rapporte le cas d'un homme qui s'était substitué au véritable témoin pour porter un faux témoignage, d'abord verbalement, puis par écrit, et qui s'était trahi au dernier moment en signant sa position de son véritable nom. Gross estimait que les faux témoins se trahissaient inévitablement, fût-ce par un seul mot, mais aussi par toute leur attitude, leur physionomie ou leurs gestes. »31 L'interprétation de Hans Gross naît véritablement à la limite de deux disciplines : la résistance détectée se trahit par les « mêmes indices », que Freud essaie de tenir à distance. Rien n'empêche qu'une intuition judiciaire puisse assigner à ces « mêmes indices » une valeur « inconsciente », qui dépasse l'établissement d'états des faits. Cette interprétation signifie ces indices-là dans une perspective psychanalytique, parce qu'ils sont le coeur de la résistance. « Vous pouvez en effet, lors de votre investigation, être induits en erreur par tel névrosé qui réagit comme s'il était coupable, bien qu'il soit innocent, parce qu'une conscience de culpabilité, toute prête et aux aguets, s'empare de l'accusation faite dans ce cas particulier. Ne prenez pas ce cas pour une invention oiseuse ; pensez à la chambre des enfants, dans laquelle on peut l'observer assez fréquemment. Il arrive qu'un enfant à qui l'on reproche un méfait dénie résolument sa culpabilité, mais pleure en même temps comme un pécheur qu'on a confondu. Vous penserez peut-être que l'enfant ment lorsqu'il assure de son innocence, mais le cas peut se présenter autrement. L'enfant n'a effectivement pas commis ce méfait-là, que vous mettez à sa charge, mais il en a en revanche commis un autre, un semblable, dont vous ne savez rien et dont vous ne l'accusez pas. Il dénie donc à bon droit sa culpabilité – pour ceci – et pourtant sa conscience de culpabilité se trahit en même temps – à cause de cela. Le névrosé adulte se comporte sur ce point – comme sur beaucoup d'autres – exactement comme un enfant ; il existe beaucoup d'êtres humains de cette sorte, et l'on peut encore se demander si votre technique réussira à différencier de tels auto-accusateurs de ceux qui sont effectivement coupables. »32 Freud tranche via la régression à un état enfantin où la responsabilité de distinguer le méfait ne passe pas uniquement par la confession de l'enfant, elle est partagée avec l'adulte. Pourquoi Hans Gross, criminologue réputé, pousse les "faits" finement vers la névrose alors que Freud est plus tranchant à propos du criminel? Il semble que l'opinion de Hans Gross penche vers une interprétation plus psychopathologique qu'un jugement criminologique. Alors que Freud se montre plus méfiant et moins enclin vers une interprétation psychopathologique. En ce sens, nous pouvons interpréter la motivation de Hans Gross comme un effort pour dispenser le névrosé d'un
32 Freud S. (1906) « Diagnostic de l'état des faits et psychanalyse » in OEuvres complètes. Psychanalyse. Tome VIII, op. cit, pp.24-25. On perçoit encore ce type de raisonnement chez dans Freud S. (1913), « Deux mensonges d'enfants » in OEuvres complètes. Psychanalyse. Tome XII, Puf, Paris, 2006. Freud présent deux cas cliniques par rapport aux mensonges des enfants. Prenons
le
deuxième
exemple
. Il s'agit d'une femme névrotique qu'à l'âge de dix ans mentait aux camarades d'école. Elle mentait en se vantant de manger la glace tous les jours car elle supposait que c'était quelque chose de distinguée. A l'occasion, elle était sensée de dessiner un cercle à main levée, mais elle triche en utilisant le compas, puis elle nie obstinément l'avoir fait, même si les preuves étaient flagrantes. Freud découvre que la fille était fortement attachée à son père : « elle fanfaronnait devant ses condisciples pour ne pas devoir diminuer son père. Lorsqu'elle apprit plus tard à traduire la glace du déjouner par "glace", la voie était alors frayée par laquelle le reproche concernant cette réminiscence pouvait déboucher dans une angoisse devant les éclats et débris de verre » (p.73). Pour nous, il peut s'agir d'un acte manqué est ses dérivations. Freud, interprète ce symptôme comme un désir incestueux masqué à l'identification au père dans la tentative de tricherie (dessiner bien comme lui). Dans le paragraphe final de l'article : « on commettrait une grave erreur si, à partir de méfaits enfantins, on pronostiquait le développement d'un caractère immorale ». Ibid., p.73. Dans cette conclusion se rejoint clairement nos propos par rapport au comportement antisocial. En effet, l'interprétation psychopathologique du comportement l'antisocial ne peut pas se passer d'une considération qui soit écartée des faits comportementales et leurs récits concomitants. Ainsi, une interprétation semble plus fructifère, et en même temps moins un jugement, si elle considère les variables métapsychologiques comme les prototypes d'une correcte observation clinique. 31 jugement qui ne soit pas de type diagnostique, tand que la réticence de Freud comme une protection de la technique psychanalytique. Suivons les faits historiques pour comprendre cette dichotomie. Il est connu que l'élément qu'il y a au milieu de celle-ci n'est pas purement technique. Juste au centre se trouve « l'affaire Gross » (Otto, fils). Ernest Jones, qui a connu Otto Gross, déclare qu'il était un homme génial mais, dans sa première lettre (13 mai 1908) à Freud, il se montre réticent pour le traitement qui Jung fait : « c'est Jung qui va le traiter sur une plan psychique et naturellement, j'en suis un peu géné car Jung a du mal à dissimuler ses sentiments et que Gross lui inspire une très nette aversion. »33 Ferenczi de son côté : « de tous ceux qui vous (Freud) ont suivi jusqu'à présent, il est le plus important. Dommage qu'il soit voué à la déchéance. »34 Jung dirait « dommage que Gross soit un tel psychopathe ; c'est une forte intelligence. »35 Freud dirait qu'il n'était « pas assez sain »36. Dans une autre lettre il dit que Gross était une « homme si précieux [] et comme residu de cette analyse [le traitement de Jung à Gross] il subsistait une relation d'amitié et de collaboration entre vous. »37 Encore que Jung repondrerait la lettre en disant : « jusqu'ici l'affaire Gross m'a consumé [] diagnostic : "démence précoce" [] en dépit de tout il est mon ami, car au fond c'est un homme bon et distingué, un esprit inhabituel. »38 Les jugements qui portent sur Gross passent par l'intelligence, l'amitié, la psychopahie, bref les qualités humaines susceptibles à être comprises comme morales. Mais ils ne sont pas suffisamment significatifs pour trancher une psychopathologie nette. La psychiatrie contemporaine pencherait pour un trouble du spectre bipolaire, alors que la clinique psychanalytique actuelle, surtout celle inspirée de Green, dirait volontiers qu'il s'agit d'un « cas limite ». Otto Gross a été trois fois interné au Burghölzli, une première fois en 1902, puis en 1904, sous les soins de Bleuler, enfin en 1908 en tant que patient de Jung. Dans une lettre datée de 10 juillet de 1902, de Bleuler à Hans Gross, le premier communiquait le diagnostic d'Otto : « Une "dépendance" compliquée d'une "constitution psychopathologique" »39. Lors de la troisième hospitalisation, Otto manifestait son rejet envers 33 Freud S., Jones E., Correspondance 1908-1939, Paris,
f, 1998, p.47. Freud S., Ferenczi S., Correspondance Tome I 1908-1914, Calman-Lévy, Paris, 1992, p.164. En fait, Ferensci dit être « enchanté » du texte Uber psychopatische Minderwertigkeiten (Infériorités psychopathiques) d'Otto Gross de 1909. Ibid., p.164. 35 Freud S., Jung C., Correspondance 1906-1914, op. cit., p.138. 36 Ibid., p.189. 37 Ibid., p.220. 38 Ibid., p.222. 39 Bair D., Jung, op. cit., p.1043. 34 32 la figure de Bleuler « Otto détestait Bleuler »40 ; puis Hans Gross demande à Freud d'intervenir pour assurer la prise en charge de son fils.
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Review: Islands of Place and Space. A Festschrift in Honour of Arne Kruse
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Donna Heddle
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English
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Spoken
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Scandinavica Vol 63 No 1 2024
Book Review: Islands of Place and
Space. A Festschrift in Honour of
Arne Kruse
Donna Heddle
University of the Highlands and Islands
DOI
10.54432/scand/MWFD5032
This work is licensed under a Creative Commons
Attribution 4.0 International License. Scandinavica Vol 63 No 1 2024 1 1 Scandinavica Vol 63 No 1 2024 CHRISTIAN COOIJMANS (ed.): Islands of Place and
Space. A Festschrift in Honour of Arne Kruse. Scottish Society for Northern Studies. Edinburgh
2022. Pp. 330. ISBN: 978-1-3999-3523-4. It is always a pleasure to review a festschrift for an esteemed colleague. Festschrifts tend to be idiosyncratic and eclectic, which makes them a
joy to read, because they encapsulate the nature of the individual
being celebrated – in this case, Dr Arne Kruse, who has recently retired
from the University of Edinburgh after a long and illustrious career. This quietly dignified and charming festschrift starts with a preface
from Angus Robertson MSP, Cabinet Secretary for the Constitution,
External Affairs, and Culture, acknowledging the importance of Arne’s
research in furthering the understanding between Scotland and
Norway. We then move to a warm and comprehensive introduction to
the life and career (and post-retirement activities) of Arne, co-written
by the editor Christian Cooijmans and Birgitte Guenier-Kruse. This
leads us to a splendid collection of fifteen contributions reflecting the
interests, academic and otherwise, the influence, and the long career
of Arne. The volume is structured particularly well, sending the reader on a
voyage of discovery flowing from one research topic to another as they
have featured in Arne’s long career, and also encapsulating other
interests, such as fishing. The order of the contributions cleverly
mirrors the relationship each contributor has had with Arne, and three
of them (Chapters III, XII, and XV) are particularly personal and
anecdotal. We begin with a paper looking at the building blocks of
language in fine academic style in an excellent paper by Arnstein
Hjelde on the changes in Minnesota-Norwegian dialects since the
1980s and we end with a lyrical prose poem about the journey home
by Hilde Rognskog and Heidi Rognskog Mella, finishing with the
emotive coda “Tomorrow, the wind will abate”, signifying journey’s
end. The sense of place and space is well established through several
papers on place names, again reflecting Arne’s interests. The second
and third contributions in the festschrift deal specifically with place
names in Norway - Botolv Helleland looks in detail at metaphors in the
mountain place names of Western Norway and Berit Sandnes looks at
the methodology surrounding the Møre and Romsdal place-name
project of 1985-95 to which Arne contributed. 1 The sixth, ninth and
eleventh contributions look at aspects of the onomastics of Scotland 2 Scandinavica Vol 63 No 1 2024 and the Norse influence on them, which became one of Arne’s great
research interests. His colleague from the Department of Scandinavian
Studies, Alan MacNiven, takes another look at – dalr place names in
Scotland; Anke-Beate Stahl analyses the specific place names of the
island of Mingulay; and Peder Gammeltoft reflect on what spatial
analysis can tell us about place names ending in -staðir and -bolstaðir. We also have fascinating papers on Norse sheilings in Caithness from
Ryan Foster, on language shift in Scotland and the Irish Sea region after
the Vikings by Pavel Iosad and on Nynorsk, which Arne enthusiastically
espoused, from Guy Puzey. Arne’s cultural heritage interests are well expressed by papers
ranging from one on Selma Lagerlöf by his long-term colleague at
Edinburgh, Bjarne Thorup Thomsen, to an explanation of the Arne
Kruse Method (being pragmatic, interdisciplinary and scholarly) in
looking at fieldwork Liv Willumsen undertook with him pertaining to
the North Berwick witch trials; to a discussion of Norse magic and Norn
words from Andrew Jennings; to a note from Brian Smith on the place
name Þursaskr which is found in Orkneyinga Saga; and to a discussion
of the herring boom and recession in Shetland from Linda Riddell. Echoing the book’s ethos, these last three contributors are islanders
themselves, writing about island matters. g
This book is certainly an analysis of islands of place and space – all
connected by the swirling, boundless intellect that is Arne Kruse. 3 3 3
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Ilustración gráfica LGTB. Caricatura, burla y erotismo
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REVISTA CHILENO-ESPAÑOLA, ACADÉMICO CIENTÍFI CA DE HUMANI DADES, ARTE Y
C U L T U R A ( I S S N : I S S N : 25 30 - 6 01 4) , N Ú M . 7 ( S E P T I E M B R E 202 0 )
ILUSTRACIÓN GRÁFICA LGTB
CARICATURA, BURLA Y EROTISMO
LGTB GRAPHIC ILLUSTRATION
CARICATURE, FUN AND EROTISM
María Aragón López
Universidad de Málaga
mariaaragon91@gmail.com
Recibido: 02 abril 2020 Aceptado: 20 mayo 2020 Publicado: 01 julio 2020
Resumen: La ilustración gráfica presenta una gran variedad de temáticas y técnicas. En
el presente artículo se estudiará en concreto la ilustración y la caricatura LGTB de carácter
burlesco y erótico haciendo un gran énfasis en las revistas satíricas de finales del siglo
XIX y principios del siglo XX, como L’Assiette au Beurre, debido a su gran relevancia y
repercusión. También veremos otros ejemplos de igual relevancia histórica y artística
como pueden ser las ilustraciones de Gerda Wegener para Douze sonnets lascifs.
Palabras clave: ilustración, caricatura, LGTB, homosexual, safismo
Abstract: The graphic illustration presents a great variety of themes and techniques. In
this article we will study specifically the LGTB illustration and caricature of a burlesque
and erotic characteristic with a great emphasis on satirical magazines from the late 19th
and early 20th centuries, like L'Assiette au Beurre, due to its great relevance and
repercussion. We will also see other examples of equal historical and artistic relevance
such as Gerda Wegener's illustrations for Douze sonnets lascifs.
Keywords: illustration, caricature, LGBT, homosexual, sapphism
Introducción
A la hora de tratar el tema de la ilustración gráfica tenemos que comprender que es
bastante amplio debido a su extensa historia, contextos y a sus diferentes técnicas y
estéticas, que han ido marcando la trayectoria y evolución de este arte. En estos últimos
años se está prestando una merecida importancia a la ilustración gráfica, con el desarrollo
MARÍA ARAGÓN LÓPEZ
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ILUSTRACIÓN GRÁFICA LGTB
ILUSTRACIÓN GRÁFICA LGTB
REVISTA CHILENO-ESPAÑOLA, ACADÉMICO CIENTÍFI CA DE HUMANI DADES, ARTE Y
C U L T U R A ( I S S N : I S S N : 25 30 - 6 01 4) , N Ú M . 7 ( S E P T I E M B R E 202 0 )
de numeroso material alrededor de esta cuestión, como artículos, libros, congresos, ferias,
etc. En definitiva, tratándola como una disciplina o categoría artística más, como puedan
ser la pintura o la escultura. Un asunto menos trabajado es el que concierne al ámbito de
la imagen gráfica LGTB, ya que, aunque también encontramos cierta información que
nos permite ahondar en dicho tema, consideramos que todavía no se ha creado una
bibliografía específica en relación a esta cuestión, sobre todo en el panorama español.
Atendiendo a esta realidad, Louis-Georges Tin afirma que «todavía está por hacer la
historia de la representación de los homosexuales a través de la caricatura» (Tin, 2012,
p.102), razón por la cual vemos necesario este trabajo de análisis sobre la ilustración
LGTB.
A continuación, nos centraremos concretamente en un conjunto de ilustraciones y
caricaturas de temática LGTB con diferentes características y objetivos como son la
sátira, la burla y el erotismo. A pesar de que muchas de las imágenes que se mencionarán
a continuación son de naturaleza despectiva, son igualmente muy interesantes respecto al
mensaje, la estética y la evolución estilística de las mismas.
La caricatura
Una caricatura, por lo general es la representación cómica, humorística o sarcástica de
uno o varios personajes o de una escena, en la que se exageran los rasgos de los
protagonistas o los mismos hechos. Tal como menciona el doctor José Enrique Peláez
Malagón (2002), la caricatura ha tenido un gran desarrollo desde el comienzo de su
existencia. Su historia comienza desde el antiguo Egipto, continúa en Grecia y en Roma,
llegando a la Edad Media y el Renacimiento, hasta que, finalmente, el uso de la litografía
y la difusión de la prensa hacen que en el siglo XIX la caricatura alcance su mayor
desarrollo, concibiéndola como «un lenguaje popular y asequible para todos» (Peláez,
2002, s. p.).
La caricatura, por tanto, debido a sus propias características, también nos puede
servir como una valiosa fuente de información. Gracias a ellas podemos conocer muchos
hechos políticos, e incluso cómo estos eran vistos por el pueblo, pues toda imagen se
enmarca en un contexto. Pero ¿qué pasa cuando la caricatura está relacionada con lo
LGTB, o más concretamente con la homosexualidad? En un primer momento, vemos
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ILUSTRACIÓN GRÁFICA LGTB
ILUSTRACIÓN GRÁFICA LGTB
REVISTA CHILENO-ESPAÑOLA, ACADÉMICO CIENTÍFI CA DE HUMANI DADES, ARTE Y
C U L T U R A ( I S S N : I S S N : 25 30 - 6 01 4) , N Ú M . 7 ( S E P T I E M B R E 202 0 )
como se usan como un instrumento para desvirtuar y difamar la figura de personajes
públicos, como reyes, políticos, militares, o cualquier persona conocida por la población,
los cuales han podido ser acusados de prácticas homosexuales y sodomitas.
Un ejemplo de lo que nos referimos es el de la propaganda pornográfica que se
usó contra los últimos reyes de Francia y su corte. Entre ellos destacamos los libelos que
hacen referencia a María Antonieta (Zweig, 2018, p.185). En dichos textos y los grabados
que los acompañan, vemos como se acusa a la reina de participar en distintas orgías y de
mantener sexo lésbico, entre otras cuestiones. Esto lo vemos en Vie de Marie-Antoinette
d‟Autriche, reine de France1 del autor Charles-Joseph Mayer y publicado en 1793. En el
grabado contemplamos como Marie-Antoinette junto con otra mujer se besan y se tocan
los genitales y los senos. Obviamente la representación de la reina y de su amante no
exagera los cuerpos o la situación, siendo por lo tanto más realista que una caricatura
como tal.
Más cerca de nuestro tiempo y espacio, continuamos viendo como esas
identidades que no se asocian con lo heteronormativo siguen siendo un motivo artístico
destacado de la caricatura. Goya nos deja un dibujo en su Álbum C el cual titula El
Maricón de la tía Gila (1808-1814). Una vez más, vemos como se incluyen personajes
de todas las clases y condiciones, en este caso más por el hecho de dejar constancia que
por crear burla. En este Álbum C nos encontramos con dibujos de carácter político,
religioso, social y económico en donde se plasman desde torturas hasta persecuciones o
escenas de toque más costumbristas, en las que podemos ver personajes de todas las
índoles, como tullidos, deformes, mendigos y demás personas monstruosas. Entre este
colectivo estaría también el “maricón” 2, perteneciendo a un grupo de personas que no
encajan con la sociedad. Esta serie de caricaturas satíricas y grotescas nos representa una
1 Título completo: Vie de Marie-Antoinette d‟Autriche, reine de France, femme de Louis XVI, roi des Français, depuis
la perte de son pucelage jusquàu 1er mai 1791. Ornée de ving-six figures et augmentée d‟une 3e partie. Avec
permission de la liberté. «Vida de María Antonieta de Austria, reina de Francia, esposa de Louis XVI, rey de los
franceses, desde la pérdida de su virginidad hasta el 1 de mayo de 1791. Adornada con veintiséis figuras y aumentada
en una tercera parte. Con permiso de la libertad.»
2 La palabra maricón vendría siendo un aumentativo de la palabra marica, y esta a su vez un diminutivo del nombre
María. En el diccionario de Covarrubias (1611) se recogía por primera vez este término en el que se entiende maricón
como hombre afeminado que se inclina a hacer cosas de mujer. Más tarde el término pasa a significar también hombre
cobarde como se recoge en el Diccionario de autoridades de 1734 en el que dice «El hombre afeminado y cobarde, y
lo mismo que Marica». No es hasta el siglo XIX que el término tiene una connotación sexual, como ocurre en
Diccionario Usual de 1899 en el que relacionan maricón con sodomita. Aunque esta expresión ha estado cargada de un
carácter peyorativo, cada día son más personas del colectivo homosexual los que la usan de manera reivindicativa como
vocablo español en sustitución del gay de origen inglés.
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ILUSTRACIÓN GRÁFICA LGTB
ILUSTRACIÓN GRÁFICA LGTB
REVISTA CHILENO-ESPAÑOLA, ACADÉMICO CIENTÍFI CA DE HUMANI DADES, ARTE Y
C U L T U R A ( I S S N : I S S N : 25 30 - 6 01 4) , N Ú M . 7 ( S E P T I E M B R E 202 0 )
realidad, en palabras de Valeriano Bozal (Bozal, 2008, p.412) «los dibujos del artista
aragonés se atienen a la más estricta individualidad y cuidan de ofrecernos con todo
esmero las características personales de cada uno de esos protagonistas, anónimos, pero
no por ello menos reales, en ni ningún caso, abstractos.» y añade más tarde (Bozal, 2008,
pp. 418-419):
Goya también se sirvió del exceso de la deformación, pero mantuvo siempre un criterio
de verosimilitud, tal como puede apreciarse, por ejemplo, en algunos de los dibujos del Álbum C,
en Este fue un cojo qe. tenia señoria, (h. 1803-24, Madrid, Museo del Prado). Éste y otros tullidos
y mendigos son figuras deformes, pero el artista aragonés, en lo cual se diferencia muy claramente
de Hogarth, los ha representado de manera que produce la sensación de que las deformidades son
propias de cada uno de ellos, y no interpretaciones o exageraciones del dibujante.
Aunque no estamos hablando de una persona deforme, las personas de otros
géneros y sexualidades se podrían enmarcar en este grupo por esa consideración de
personas ambiguas, invertidas o desviadas. Pero lo interesante es más bien la
representación misma del personaje que nos muestra Goya.
La burla
Siguiendo con esa mirada de la sociedad que juzga los géneros no normativos y las
sexualidades indeterminadas, encontramos un libro que nos ayuda a explicar estos
fenómenos en donde lo distinto, en esta ocasión en relación con lo femenino, se hace
patente en el arte. Nos referimos a la obra de Nicole G. Albert, Lesbian Decadence:
Representations in Art and Literature of Fin-de-Siècle France. G. Albert, realiza una
recopilación de textos históricos y científicos, novelas, poemas, pinturas e ilustraciones
en relación con el safismo, analizándolo como un fenómeno social de finales del siglo
XIX en Francia, que se considera como su época dorada. Debido a que en estos
momentos la homosexualidad masculina se había convertido en un gran tabú, tanto el
mundo de las artes y las letras como el mundo científico se centraron en la
homosexualidad femenina, ya sea para estudiarla, ridiculizarla por ser contranatural o
tratarla como tema erótico. Lo que no cabe duda es que el tema del safismo gozó de gran
popularidad llegando incluso a rozar la obsesión.
De esta forma, en Lesbian Decadence vemos un recorrido por todos esos textos e
imágenes las cuales nos contextualizan este fenómeno como la poetisa Safo de Lesbos,
MARÍA ARAGÓN LÓPEZ
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ILUSTRACIÓN GRÁFICA LGTB
ILUSTRACIÓN GRÁFICA LGTB
REVISTA CHILENO-ESPAÑOLA, ACADÉMICO CIENTÍFI CA DE HUMANI DADES, ARTE Y
C U L T U R A ( I S S N : I S S N : 25 30 - 6 01 4) , N Ú M . 7 ( S E P T I E M B R E 202 0 )
la cual fue estigmatizada por su condición, retratándola con numerosos amoríos y como
mujer promiscua. También encontramos referencias sobre Les Fleurs du Mal de Charles
Baudelaire o diversos estudios, entre ellos, los que relaciona el amor entre mujeres con el
vicio y la enfermedad mental 3, creando verdaderamente una mitología decadente de la
lesbiana. Así pues, aunque no se criminalizaba estas acciones amorosas, sí eran señaladas
como inmoral por muchos.
Muchas de las ilustraciones que se recogen en Lesbian Decadence muestran
caricaturas de este hecho por algunas de las revistas francesas de la época como L’Assiette
au Beurre, Le Rire o Fantasio, las cuales dedicaban en sus distintos espacios de las
revistas a la homosexualidad, por lo que podemos llegar a la conclusión de que era un
tema muy frecuente en estas revistas satíricas. En concreto, L’Assiette au Beurre, aunque
de vez en cuando tratase dicho tema, dedicó exclusivamente dos números a la
homosexualidad. El primero de estos números fue el 422 publicado el 1 de mayo de 1909,
donde dedicaban un capítulo especial a los P’tits jeun’hommes, es decir, a los jovencitos.
Esos hombres considerados amanerados y ridiculizados con estas caricaturas satíricas
donde aparecen maquillados y con posturas seductoras.
Posteriormente, en el número 568 que salió el día 2 de marzo de 1912 se dedicó
al tema del safismo. En ella retrata algunos de los hechos más característicos de estas
prácticas de forma sarcástica. Son las Mesdames, Messieurs la propia expresión denota
esa ambigüedad en cierto tono burlesco. En la portada de esta revista satírica nos muestra
un personaje mitad mujer mitad hombre, es decir mujeres vistiendo y comportándose a la
manera masculina. Siguiendo este estilo de representación, son muchas las mujeres que
son retratadas de esta forma, vestidas con trajes, sombrero, fumando y con posturas
dominantes, lo que vendría a ser en términos más despectivos marimacho. Básicamente
en ambos números podemos ver una serie de clichés que se repiten y se asocian a la
homosexualidad masculina y a la homosexualidad femenina.
La diferencia entre estos dos números y las caricaturas e ilustraciones de otras
revistas es que las representaciones de los afeminados tienen como objetivo principal el
de restar valor a los hombres de los que hablan ya sea a nivel general o concretando en
una figura de poder como puede ser un personaje político. De forma opuesta, las mujeres
5
3 Ambas cuestiones se tratan en la segunda parte del libro Her Traits, Her Vices, and Her Sexual Aberrations.
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ILUSTRACIÓN GRÁFICA LGTB
ILUSTRACIÓN GRÁFICA LGTB
REVISTA CHILENO-ESPAÑOLA, ACADÉMICO CIENTÍFI CA DE HUMANI DADES, ARTE Y
C U L T U R A ( I S S N : I S S N : 25 30 - 6 01 4) , N Ú M . 7 ( S E P T I E M B R E 202 0 )
que aparecen virilizadas en estas revistas satíricas no tienen una connotación tan negativa.
De hecho, aunque se ha comentado que siempre hay cierto toque burlesco, estas
señora’señores (Tin, 2012, p.105) no supondrían una amenaza real según muchos
dibujantes, que así lo reflejaban en sus ilustraciones, haciendo énfasis en esta cuestión.
Muestra de ello, lo encontramos en una caricatura de la revista, en el texto vemos que un
hombre le increpa a otro Et vous tolérez que votre femme… avec son amie… a lo que
contesta Bah! c’est ma façon à moi de n’être pas cocu!4 El principal objetivo de estas
caricaturas era satirizar ciertas cuestiones que empiezan a surgir en el cambio de siglo,
como los cabarets homosexuales o los bares para lesbianas, que irrumpían en mitad de
una sociedad conservadora, dando lugar incluso a que muchas de estas ilustraciones
fuesen muy hirientes para este colectivo o para personas específicas.
Este último caso lo podemos encontrar en lo que se conoció como el escándalo
Harden-Eulenburg, que sucedió durante el gobierno del emperador Guillermo II de
Alemania, quien estuvo implicado en el mismo. Aunque los motivos no están del todo
claros, se cree que algunas personas, entre ellas Maximilian Harden, fundador de la
revista Die Zukunft, estaban descontentas con las políticas externas del gobierno del
káiser, desatando una serie de críticas en los periódicos y atacando a la vida privada del
círculo más próximo de Guillermo II, e incluso al propio emperador. Se divulgaron
artículos y viñetas en donde estos personajes eran señalados por haber experimentado
prácticas homosexuales de todo tipo (Tamagne, 2003, pp. 49). Obviamente, y dentro del
marco que venimos presentando, podemos interpretar la acusación de homosexualidad de
los dirigentes de un país como un intento de desprestigiar las acciones políticas de estos
y quitarles toda su autoridad. La acusación de homosexualidad a su vez asociada con la
incapacidad de estos de decidir cuestiones políticas y bélicas por su carácter amanerado
y femenino.
No solo los periódicos alemanes se hicieron eco de estos escándalos, pues como
menciona Steakley (1983), el escándalo llegó a muchos otros países europeos:
The Eulenburg Affair was of such consuming interest to France that it remains the only
country to have produced monographs on the subject. Switzerland and Belgium tried to maintain
4«-Y toleráis a vuestra esposa… con su amiga…
MARÍA ARAGÓN LÓPEZ
-¡Bah! ¡Es mi manera de no ser cornudo!»
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ILUSTRACIÓN GRÁFICA LGTB
REVISTA CHILENO-ESPAÑOLA, ACADÉMICO CIENTÍFI CA DE HUMANI DADES, ARTE Y
C U L T U R A ( I S S N : I S S N : 25 30 - 6 01 4) , N Ú M . 7 ( S E P T I E M B R E 202 0 )
their neutrality, but cartoons from these countries show the opprobrium they attached to
homosexuality. In contrast, the Italian treatment tended more clearly to the sensational (pp.36)5.
En el caso francés se producen monográficos sobre el asunto, y las revistas
satíricas, entre ellas Fantasio, Le Rire o L’Assiette au beurre, rápidamente reproducen las
viñetas de la prensa alemana y crean otras caricaturas mofándose del tema dejando la
dignidad y el honor del pueblo alemán defenestrado. En el caso de la revista antes
mencionada, L’Assiette au beurre, vuelve a dedicar uno de sus números al tema de la
homosexualidad, en este caso en relación con la controversia del caso Harden-Eulenburg
y de los militares alemanes. En el número 377 del 20 de junio de 1908 podemos ver ya
en la portada como el emperador se pregunta en dónde ha quedado el prestigio de su
imperio, y si seguimos por las páginas del mismo continuamos viendo una serie de
imágenes y caricaturas aludiendo al mismo problema, sugiriendo también la incapacidad
de estas personas de tomar decisiones por su condición.
El erotismo
Por otro lado, no todas las muestras de artes que encontramos de esta época están
relacionadas con la sátira, la burla y la denigración. En el caso del amor sáfico también
encontramos visiones en donde la pareja femenina tiene un encanto seductor para los
espectadores. Es un nuevo comienzo de la imagen de las lesbianas donde se rechaza esas
caricaturas varoniles siendo elevado el safismo a un motivo realmente artístico. Una vez
más, dentro de Lesbian Decadence encontramos varios ejemplos en donde estas
relaciones amorosas se reflejan desde esta otra perspectiva. Una muestra de lo que
decimos lo encontramos en el libro Femme honnêtes! de Joséphin Péladan (1885).
Ilustrado por Fedinand Bac, muestra como las amantes Lucie y Berthe se abrazan entre
el cariño, la sensualidad y la pasión, justo antes de besarse.
De una manera más explícitamente sexual, nos encontramos la obra de Franz von
Bayros, un artista que destacó por sus ilustraciones eróticas, las cuales fueron censuradas
«El asunto Eulenburg fue de un interés tan grande para Francia que sigue siendo el único país que ha producido
monografías sobre el tema. Suiza y Bélgica intentaron mantener su neutralidad, pero las caricaturas de estos países
muestran el oprobio que atribuían a la homosexualidad. En contraste, el tratamiento italiano tendió más claramente a
lo sensacional.» (Steakley 1983, pp. 36)
5
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ILUSTRACIÓN GRÁFICA LGTB
REVISTA CHILENO-ESPAÑOLA, ACADÉMICO CIENTÍFI CA DE HUMANI DADES, ARTE Y
C U L T U R A ( I S S N : I S S N : 25 30 - 6 01 4) , N Ú M . 7 ( S E P T I E M B R E 202 0 )
o prohibidas, obligándole incluso a abandonar el país en donde se encontraba. Su trabajo
más controvertido fue el que realizó en 1911 titulado como Erzahlungen vom
Toilettentisch (Tales from the Dressing Table)6, y como es de esperar, entre estas
ilustraciones encontramos también esas relaciones entre mujeres que disfrutan del placer
sáfico desde un punto de vista más masculino, pero igualmente bello, delicado y con
cierto gusto al decadentismo.
Otro punto de vista, erótico, seductor y elegante, es el que nos muestra la artista e
ilustradora Gerda Wegener 7. Aparte de ser conocida por sus obras, también destaca por
estar casada con el primer transexual de la historia, o por lo menos de los primeros
hombres conocidos que se sometieron a una cirugía para cambiar su género de forma
oficial. Se trataba de otro artista, Einar Wegener, quien adquirió después la identidad de
Lili Elbe. Al contrario de lo que pudiéramos pensar, Gerda, más allá de condenar la
transformación de su marido al sexo opuesto, se aleja de los prejuicios de su época y
ayuda a que la transformación se produzca. Este puede ser quizás uno de los motivos de
su interés de representar esas sexualidades diferentes y fuera de lo heteronormativo,
habiéndose afirmado al respecto que «la revendication implicite ou explicite des droits
des minorités liées à leur identité sexuelle est partout presente dans les illustrations, les
dessings et les peintures de Gerda Wegener, dès les années 1900» 8 (Claustrat, 2015, s.
p.). Ya sea un acto reivindicativo implícito o explicito, si es verdad que se ve interesada
por mostrar la variedad desde un punto de vista humano y sensible.
De esta manera, Gerda Wegener no solo destaca por la representación de aquellas
temáticas dirigidas expresamente a los hombres, como puede ser la guerra, sino que
también expresa en sus dibujos y pinturas los temas pocos tratados por la sociedad por
estar relacionadas con esas minorías rechazadas por su condición o identidad sexual,
como pueden ser las lesbianas, los transexuales y travestidos. Así pues, comenzamos a
ver retratos de Lili mostrando esa nueva identidad, como si fuese la musa del artista,
representándola con tocados distintos, vestimentas de todos los colores, acompañada por
la propia artista e incluso desnuda. Eva María Ramos Frendo comenta sobre este hecho
6 «Cuentos desde el tocador».
7 Gerda Wegener (1885-1904) fue una artista danesa conocida principalmente por los retratos y las ilustraciones con
carácter satírico para libros y revistas que realizó cuando residía en París.
8 «la reivindicación implícita o explícita de los derechos de las minorías relacionadas con su identidad sexual se presenta
en todas parte de las ilustraciones, dibujos y pinturas de Gerda Wegener, desde los años 1900»
MARÍA ARAGÓN LÓPEZ
8
MARÍA ARAGÓN LÓPEZ
ILUSTRACIÓN GRÁFICA LGTB
ILUSTRACIÓN GRÁFICA LGTB
REVISTA CHILENO-ESPAÑOLA, ACADÉMICO CIENTÍFI CA DE HUMANI DADES, ARTE Y
C U L T U R A ( I S S N : I S S N : 25 30 - 6 01 4) , N Ú M . 7 ( S E P T I E M B R E 202 0 )
que «Lili será continuamente el disfraz o máscara bajo el que se oculta Einar o, más bien,
podríamos decir que es ese disfraz el que hace posible a Einar desenmascarar su verdadera
condición sexual en la figura de Lili» (Ramos, 2016. s. p.).
Como hemos dicho anteriormente, Gerda Wegener no destacó solo por sus
retratos y pinturas, sino que también fue muy reconocida por sus ilustraciones y
caricaturas. Entre ellas destacamos las que realizó para Douze sonnets lascifs 9 de Louis
Pearceau. A pesar de su estilo Art Decó, de la gran calidad de las imágenes, de la
elegancia de las formas y de la sutileza de los colores, nos muestra unas acuarelas donde
aparecen unas mujeres delicadas remarcadas por óvalos y círculos en situaciones
sexualmente explicitas. En contraposición con Franz von Bayros, es la mirada femenina
la que predomina aquí, y a pesar de los tintes eróticos, no podemos negar la delicadeza y
dulzura con la que se muestran estas ilustraciones.
Conclusión
Tras el análisis de los diversos ejemplos expuestos, no podemos negar que el hecho de
encontrar tal cantidad de ilustraciones de temática homosexual, ya sea desde un punto de
vista satírico o de carácter erótico, es porque realmente nos encontramos ante un
fenómeno social, que no solo abarcó a las ilustraciones de las revistas y libros, sino que
llegó a otros ámbitos como la literatura y al panorama científico.
Igualmente, también podemos destacar como la forma de representación y las
características que encontramos en las ilustraciones mencionadas difieren entre las
sarcásticas y las eróticas, pues cada una es propia de un contexto determinado o persigue
objetivos bien distintos. Por un lado, unas resaltan los clichés, llevándolos al absurdo,
mientras que las otras resaltan la sensualidad y la lujuria. Pero, ante todo, ambas son el
reflejo de una misma época en la que poco a poco el mundo LGTB iba apareciendo en el
panorama social y político para convertirse en un tema que a nadie dejaba indiferente.
9
9 «Doce sonetos lascivos»
MARÍA ARAGÓN LÓPEZ
MARÍA ARAGÓN LÓPEZ
ILUSTRACIÓN GRÁFICA LGTB
ILUSTRACIÓN GRÁFICA LGTB
REVISTA CHILENO-ESPAÑOLA, ACADÉMICO CIENTÍFI CA DE HUMANI DADES, ARTE Y
C U L T U R A ( I S S N : I S S N : 25 30 - 6 01 4) , N Ú M . 7 ( S E P T I E M B R E 202 0 )
Recursos bibliográficos
Albert, Nicole G. (2016). Lesbian Decadence: Representations in Art and Literature of
Fin-de-siècle France, Translated by Nancy Erber and William A. Peniston, Harrington
Park Press, New York.
Bozal, Valeriano (2008). ‘Dibujos grotescos de Goya’. En Anales de Historia del Arte,
Universidad complutense de Madrid, pp. 407-426.
Martínez Moro, Juan (2004). La ilustración como categoría: Una teoría unificada sobre
arte y conocimiento, Ediciones Trea, S.L., España.
Peláez Malagón, José Enrique (2002), Historia de la caricatura. Clío: History and History
Teaching, Nº 27. Recuperado de: (http://clio.rediris.es/arte/caricaturas/caricatura.htm)
Ramos Frendo, Eva María (2016). Las ilustraciones de la danesa Gerda Wegener (18841940) en el semanario francés La Baionnette. Una satírica visión femenina de la Primera
Guerra Mundial. AACADigital: revista de la Asociación Aragonesa de Críticos de Arte,
Nº 34. Recuperado de: (http://www.aacadigital.com/contenido.php?idarticulo=1192)
Steakley, James D. (1983). Iconography of a Scandal: Political Cartoons and the
Eulenburg Affair. Studies in Visual Communication, Volume 9, Issue 2, University of
Pennsylvania,
pp.
20-51.
Recuperado
de:
(https://repository.upenn.edu/cgi/viewcontent.cgi?referer=&httpsredir=1&article=1211
&context=svc)
Tamagne, Florence (2003). ‘Caricatures homophobes et stéréotypes de genre en France
et en Allemagne: la presse satirique, de 1900 au milieu des années 1930’. En Le Temps
des
médias,
Nouveau
Monde
éditions,
pp.
42-53.
Recuperado
de:
(https://www.cairn.info/revue-le-temps-des-medias-2003-1-page-42.htm)
Tin, Louis-Georges (2012). Diccionario de la homofobia, Editorial Akal, Madrid,
España.
10
MARÍA ARAGÓN LÓPEZ
MARÍA ARAGÓN LÓPEZ
ILUSTRACIÓN GRÁFICA LGTB
ILUSTRACIÓN GRÁFICA LGTB.
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tel-03850840-2022UPSLO002_archivage.txt_4
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French-Science-Pile
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Open Science
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Various open science
| 2,022
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Mechanisms of star formation quenching in local galaxies. Astrophysics [astro-ph]. Université Paris sciences et lettres, 2022. English. ⟨NNT : 2022UPSLO002⟩. ⟨tel-03850840⟩
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None
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English
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Spoken
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2001, ApJ, 556, 121 Kewley, L. J., Nicholls, D. C., & Sutherland, R. S. 2019, ARA&A, 57, 511 Koss, M., Mushotzky, R., Treister, E., et al. 2011, ApJ, 735, L42 Krajnović, D., Cappellari, M., de Zeeuw, P. T., & Copin, Y. 2006, MNRAS, 366, 787 Lacy, M., Baum, S. A., Chandler, C. J., et al. 2020, PASP, 132, 035001 Lahén, N., Johansson, P. H., Rantala, A., Naab, T., & Frigo, M. 2018, MNRAS, 475, 3934 Mannucci, F., Cresci, G., Maiolino, R., Marconi, A., & Gnerucci, A. 2010, MNRAS, 408, 2115 Maschmann, D., & Melchior, A.-L. 2019, A&A, 627, L3 Maschmann, D., Melchior, A.-L., Mamon, G. A., Chilingarian, I. V., & Katkov, I. Y. 2020, A&A, 641, A171 McAlpine, S., Harrison, C. M., Rosario, D. J., et al. 2020, MNRAS, 494, 5713 Mendenhall, W., & Sincich, T. 2011, A Second Course in Statistics Regression Analysis (Pearson Education) Mesa, V., Duplancic, F., Alonso, S., Coldwell, G., & Lambas, D. G. 2014, MNRAS, 438, 1784 Nevin, R., Blecha, L., Comerford, J., & Greene, J. 2019, ApJ, 872, 76 Osterbrock, D. E., & Ferland, G. J. 2006, Astrophysics of Gaseous Nebulae and Active Galactic Nuclei (Sausalito, CA: University Science Books) Patton, D. R., Torrey, P., Ellison, S. L., Mendel, J. T., Scudder, J. M. 2013, MNRAS, 433, L59 Pawlik, M. M., Taj Aldeen, L., Wild, V., et al. 2018, MNRAS, 477, 1708 Poggianti, B. M., & Wu, H. 2000, ApJ, 529, 157 Prieto, J., Escala, A., Privon, G., & d’Etigny, J. 2021, ArXiv e-prints [arXiv:2101.09407] Saintonge, A., Kauffmann, G., Wang, J., et al. 2011, MNRAS, 415, 61 Sakamoto, K., Aalto, S., Combes, F., Evans, A., & Peck, A. 2014, ApJ, 797, 90 Salim, S., Lee, J. C., Janowiecki, S., et al. 2016, ApJS, 227, 2 Schawinski, K., Thomas, D., Sarzi, M., et al. 2007, MNRAS, 382, 1415 Shah, E. A., Kartaltepe, J. S., Magagnoli, C. T., et al. 2020, ApJ, 904, 107 Sharma, R. S., Choi, E., Somerville, R. S., et al. 2021, ArXiv e-prints [arXiv:2101.01729] Solomon, P. M., Downes, D., Radford, S. J. E., & Barrett, J. W. 1997, ApJ, 478, 144 Somerville, R. S., & Davé, R. 2015, ARA&A, 53, 51 Strauss, M. A., Weinberg, D. H., Lupton, R. H., et al. 2002, AJ, 124, 1810 Tubín, D., Treister, E., D’Ago, G., et al. 2021, ApJ, 911, 100 van Dokkum, P. G., Nelson, E. J., Franx, M., et al. Facteur de conversion CO/H2
Calibrer la relation entre l’intensité observée de CO et la masse de gaz H2 est crucial pour interpréter la quantité de gaz susceptible de former des étoiles. Une relation répandue est: Mmol = αCO LCO (4.16) où Mmol est exprimée en masses solaires M⊙ et LCO en K km s−1 pc2. αCO est alors un rapport masse-lumière. Ce rapport dépend de différents paramètres, tels que la densité du gaz, la température et la métallicité. La valeur de α = 4, 36 ± 0, 9 M⊙ /(K km s−1 pc2, qui tient compte de la quantité d’Helium, est souvent utilisée pour la Voie Lactée, les galaxies proches et celles à faible métallicité (par ex. Bolatto et al., 2008; Schinnerer et al., 2010). Une valeur de ce facteur qui tient compte de la métallicité est discutée dans divers travaux (Genzel et al., 2015; Tacconi et al., 2018). Observations à l’IRAM 30m
Afin d’estimer le contenu en gaz moléculaire des galaxies DP/MaNGA et de mesurer leur efficacité à former des étoiles, nous avons observé ces 29 sources avec le télescope 30m de l’Institut de Radio Astronomie Millimétrique (IRAM). La luminosité du CO est donnée par l’équation 5.4 et la masse totale de gaz moléculaire est obtenue avec l’équation 5.5. J’ai appliqué une correction d’ouverture pour les galaxies étendues au-delà de la taille du faisceau primaire (22′′ à la fréquence de la transition CO(1-0)), afin de tenir compte du gaz moléculaire présent dans les 91
Exploring the cold gas content of the DP/MaNGA galaxies
In this chapter, I present the observations I conducted at the IRAM 30m telescope and at the Nançay Radio Telescope to respectively obtain molecular and atomic gas content within the DP/MaNGA sources. First I give brief introductions to what can be inferred from cold gas observations, then I describe the setups of the observations and finally I present the results and analyse them in the light of other surveys of nearby galaxies. CHAPTER 5. EXPLORING THE COLD GAS CONTENT OF THE DP/MANGA GALAXIES 1 M
olecular gas phase In the typical conditions within the interstellar medium, the H2 molecule cannot be observed directly because of very small rotational transition probabilities and thus weak line emission, due to its lack of an electric dipole moment. Thus it is necessary to use other molecules in order to trace H2 by making assumptions on the relation between the abundance of the observed molecule and the one of H2. The most commonly tracer of H2 is the carbon monoxyde CO. This molecule becomes excited through collisions with the H2 molecule and its de-excitation results in the emission of photons in observable wavelength regions. The most studied CO rotational transitions are CO(J 1 → 0) and CO(J 2 → 1) that are at the heart of various molecular gas surveys and that are commonly referred to as CO(1-0) and CO(2-1). Their respective rest-frame frequencies are νCO(1−0) = 115.271 GHz and νCO(2−1) = 230.538 GHz. 1.1 CO-to-H2 conversion factor
Calibrating the relation between the observed CO intensity and the total H2 gas mass is crucial to interpret the amount of gas that is likely to form stars. A standard methodology gives: N(H2 ) = XCO ICO (5.1) where N(H2 ) is the dihydrogen column density in cm−2 and ICO is the observed CO integrated line intensity in K km s−1. The factor XCO is thus given in cm−2 /(K km s−1 ). By integrating over the emitting area and correcting by the mass contribution of heavier elements, another relation is adopted: Mmol = αCO LCO (5.2) where Mmol is given in units of M⊙ and LCO in K km s−1 pc2. With this definition, αCO is a mass-to-light ratio. One refers to “CO-to-H2 conversion factor” for both XCO and αCO. This factor depends on different environmental parameters such as the gas density, temperature and metallicity. In the Milky Way, as well as in nearby star-forming and low-metallicity galaxies, a value of α = 4.36 ± 0.9M⊙ /(K km s−1 pc2 ) accounting for helium has been found using different estimate methods (e.g. Dame et al., 2001; Bolatto et al., 2008; Schinnerer et al., 2010; Leroy et al., 2011). At higher redshift, the metallicity dependence of the factor can be taken into account through (as in Genzel et al., 2015; Tacconi et al., 2018): αCO = α q 0.67 × exp(0.36 · 108.67−log Z ) × 10−1.27·(log Z−8.67) (5.3) where α is the factor given above and where log Z=12+log(O/H), the gas-phase metallicity based on the Pettini & Pagel (2004) calibration. 2 Observations at IRAM 30m
We aimed at measuring the molecular gas content of the DP/MaNGA galaxies in order to estimate their star formation efficiency and study the process of quenching within these systems. We observed all objects with the IRAM 30m telescope at Pico Veleta in Spain. 2. OBSERVATIONS AT IRAM 30M Data acquisition and reduction
We used the spectral line receiver Eight MIxer Receiver (EMIR) in the configuration E0/2, which enables us to simultaneously operate in the 3 and 1.3 mm windows. The EMIR bands were connected to two backends: the Fast Fourier Transform Spectrometers (FTS) and the Wideband Line Multiple Autocorrelator (WILMA), allowing us to obtain measurements with a resolution of 0.195 MHz and 2 MHz, respectively. We used the symmetrical wobbler switching mode, in which the secondary mirror moves up to ±120 arcsec in azimuth. Since the telescope are performed with the on-off mode enabled by the rapid motion of the secondary mirror, the on and off positions are shifted along the azimuthal axis and therefore the airmass does not vary between both scans. The provided baselines are thus better than in the position switching mode for instance. Scans of 6 min duration were performed and encompassed on-source and off-source sub-scans of 30 s duration each. Pointing calibrations were taken on a nearby planet or bright quasar. Telescope focusing was regularly carried out. Integrations were repeated until a signal was detected and a reasonable signal-to-noise ratio of the line was reached. Multiple integrations were combined in the dedicated software package class. Usually, baselines were calculated in line-free regions using a linear fit, except for some cases in which we evaluated that higher order polynomials were required, based on visual inspection. Scans from September 1st, 2019, were discarded as the weather conditions were bad on this day. The averaged spectra were smoothed by binning the velocity channels to a velocity resolution of ∼ 64 km s−1. The reduced spectra are displayed in Figs. 5.1 and 5.2, with the Legacy survey snapshot to see the MaNGA coverage as well as the IRAM 30m telescope beam-widths. 2.2 Molecular gas analysis
The CO luminosity is expressed as: LCO K km s−1 pc−2 3.25 × 107 = (1 + z) FCO Jy km s−1 νrest GHz −2 DL Mpc 2 (5.4) where FCO is the velocity-integrated flux, νrest is the rest-frame frequency and DL the luminosity distance. For all detected galaxies, we compute the molecular gas mass MH2 following: MH2 = αCO LCO (5.5) where αCO is the estimated CO-to-H2 conversion factor. For the galaxies which are extended over a region broader than the beam size (22′′ at the frequency of CO(1-0)), an aperture correction has to be applied in order to account for the molecular gas in the non-covered regions. I follow the procedure explained in Lisenfeld et al. (2011). An exponential distribution is assumed for CO (CO maps have shown that such distributions describe the CO emission well, e.g. Nishiyama et al., 2001; Regan et al., 2001; Leroy et al., 2008). I consider the optical radius at the 25 mag isophote, r25, by dividing the optical diameter at the 25 mag isophote extracted from the NASA/IPAC Extragalactic Database (NED)1. We can assume re /r25 = 0.2 where re is the CO scale length (Lisenfeld et al., 2011). I measure the galaxy inclination using the minor-tomajor axial ratio b/a given in the MaNGA PyMorph photometric value-added catalogue. The ratio is estimated based on a 2D surface brightness fit, either with a Sérsic profile or with a 1 https://ned.ipac.caltech.edu/
CHAPTER 5. EXPLORING THE COLD GAS CONTENT OF THE DP/MANGA GALAXIES
Sérsic+Exponential profile (Fischer et al., 2019). I compute the inclination i as: cos i = s (b/a)2 − q02, 1 − q02 (5.6) where q0 describes the intrinsic axial ratio of an edge-on observation and is set to q0 = 0.2 (Holmberg, 1958, see also e.g. Tully et al. (2009)). Following Lisenfeld et al. (2011), I estimate the correction factor: fa = πre2 (Z 0 ∞ dx Z ∞ 0 dy exp −ln(2) " 2x ΘB 2 2 y cos(i) ΘB + exp − p x2 + y 2 re 2 #!! )−1 (5.7), where ΘB is the FWHM of the telescope beam, at the observed frequency. The integration of Eq. 5.7 is carried out numerically. The correction factor depends on the scale length of the CO emission, the beam size and the inclination of the galaxy. The correction factors obtained for the DP/MaNGA galaxies are such that fa ≤ 2. 2. OBSERVATIONS AT IRAM 30M
ID r25 i corr. factor αCO M⊙ /(K km s− 1 pc2 ) MH2 109 M⊙ μH2 log(SFE) yr−1 tdepl Gyr 1.81 2.00 1.13 1.23 1.24 1.59 1.52 1.53 1.45 1.17 1.22 1.73 1.15 1.36 1.15 1.31 1.21 1.36 1.25 1.18 1.46 1.22 1.20 1.23 1.14 1.12 1.15 1.19 1.09 3.9 3.9 3.8 3.8 3.8 4.2 3.9 4.0 4.0 3.8 3.8 4.2 3.9 3.9 3.8 3.8 3.9 4.1 3.8 3.9 4.1 4.1 4.1 4.0 3.8 4.1 4.1 4.1 4.0 1.3 2.3 0.3 0.6 5.5 3.2 1.0 3.7 4.1 4.4 2.1 4.4 6.1 3.1 3.3 1.5 3.5 10.6 7.5 9.9 13.9 11.8 9.7 43.0 17.2 30.0 13.2 32.4 31.8 -1.8 -1.3 -1.6 -1.7 -0.9 -1.6 -1.8 -1.3 -1.5 -0.9 -1.3 -1.5 -1.1 -1.4 -1.4 -1.5 -1.4 -1.1 -1.1 -1.1 -0.9 -0.9 -1.0 -0.6 -0.9 -0.6 -1.0 -0.6 -0.7 -9.1 -9.3 -8.9 -9.4 -9.3 -9.1 -9.4 -9.0 -9.3 -9.3 -8.9 -9.3 -8.8 -8.8 -8.4 -9.4 -9.0 -9.4 -9.3 -8.6 -9.4 -9.8 -9.8 -9.4 -8.7 -9.6 -9.1 -9.7 -9.1 1.3 2.1 0.9 2.6 2.0 1.3 2.4 1.1 1.9 1.8 0.8 2.1 0.6 0.7 0.3 2.3 0.9 2.5 1.9 0.4 2.7 5.8 6.9 2.8 0.5 4.2 1.4 4.7 1.1 ◦ G1 G2 G3 G4 G5 G6 G7 G8 G9 G10 G11 G12 G13 G14 G15 G16 G17 G18 G19 G20 G21 G22 G23 G24 G25 G26 G27 G28 G29 30.495 45.505 14.56 15.415 18.935 27.495 21.78 29.765 24.685 17.01 17.265 38.99 14.955 22.19 14.665 18.765 16.29 20.76 17.265 14.085 20.33 17.88 13.69 13.74 13.71 12.01 10.895 13.875 8.81 33 67 75 41 66 46 0.0 63 53 76 58 71 67 56 66 44 55 47 49 48 65 35 0.0 60 53 0.0 39 32 97
Table (5.1) Relevant properties of the DP/MaNGA galaxies to explore their molecular gas content. THe first column gives an arbitrary identifier to each galaxy; columns 2 to 4 display the radius at the 25 mag isophote, the inclination and the correction factor for aperture effect. Columns 5 to 9 refer to inferred quantities from the CO observations: the CO-to-H2 conversion factor, the H2 mass, the H2 fraction, the logarithmic star formation efficiency and the depletion time. I compute the conversion factor αCO following Eq. 5.3. The mean conversion factor for the 29-galaxy sample is 3.9 with a standard deviation of 0.1. Star-formation main sequence I am able to investigate how some properties derived from the CO observations vary across the SFMS. To do this and to get homogeneous values with current CO surveys on MaNGA galaxies, I make use of the measurements found in Salim et al. (2016), if available, otherwise 98
CHAPTER 5. EXPLORING THE COLD GAS CONTENT OF THE DP/MANGA GALAXIES
in Brinchmann et al. (2004). I define the molecular gas fraction μH2 = MH2 /M⋆. I measure a median molecular gas fraction for the DP/MaNGA sample of μH2 = 0.07, computed with the detected sources. I also estimate the star formation efficiency SFE = SFR/MH2, expressed in yr−1, and the depletion time tdepl which is the inverse of SFE. The latter quantity is an indicator of the necessary time to convert all the available molecular gas into stars at the current star formation rate. The median star formation efficiency is log(SFE/yr−1 ) = −9.3.
Figure 5.3
shows the offset
with respect to the star-formation main sequence defined by Whitaker
et al
. (2012), for the 29 DP/MaNGA galaxies.
I
colour
-coded them
according to the classification detailed
in
1.1.
The left panel of Fig. 5.4 shows the galaxies in the SFR-M⋆ plane, with a colour-coding referring to the molecular gas fraction. The right panel displays the SFE as colour-coding. Through the correlator, these signals are then multiplied and time averaged. The resulting output signal is: V2 Rc = cos(w τg ) (5.10) 2 As an even function, the correlator only detects the even part of the brightness distribution of the observed source. It is possible to introduce a second correlator which adds a π/2 phase delay to the output of one of the antennae. The second correlator has then as output signal: 2 Rs = V2 sin(w τg ). Both correlators can be combined into a complex value, that is referred to as a visibility V(u,v): Z V (u, v) = Rc − i Rs = u·s I(s) e−2 π i ν c ds (5.11) where I is the spatial sky brightness distribution. An interferometer thus gives a measure of the two dimensional Fourier transform of the spatial sky surface brightness distribution I. This distribution is retrieved by applying an inverse Fourier transform on the measured visibilities. The more antennae composing the interferometer, the better the (u, v) plane is sampled. 3.2 Proposed project
The merging system MaNGA ID 1-114955 that we describe in the previous chapter presents a peculiar stage of pre-coalescence merger with strong kinematic disturbances. As we detected a large reservoir of molecular gas in this system with the IRAM 30m telescope, this system probably gained a large part of its gas mass through the merging process. By mapping this galaxy with the NOEMA telescope, we will be able to spatially resolve the molecular gas and to conclude on its dynamics. We will be able to disentangle the molecular gas content corresponding to the stellar components of both galaxies. The mapping of both CO transitions will enable us to investigate the excitation and the density of the gas, using the CO(2-1)/CO(1-0) ratio. In addition, we expect a continuum flux of 30 μ Jy and 12 μ Jy at 3 mm and 1 mm respectively and a high resolution map might provide new arguments on the relation between the radio continuum flux and the star formation in a complex system, and possibly the presence of a hidden AGN. We will distinguish possible point sources at the centre of each galaxy, and separate them from diffuse emission. We already have measurements of the CO(1-0) and CO(2-1) gas content with the IRAM 30m telescope within a beam of 23′′ and 12′′ in diameter respectively. The respective measured velocity-integrated flux and linewidth are FCO(1−0) = 27Jy km s−1, FWHMCO(1−0) = 624km s−1 and FCO(1−0) = 117Jy km s−1, FWHMCO(2−1) = 897km s−1. Since the molecular gas in galaxies is not equally distributed, we expect the signal to be dispatched into clumps and streams. We thus assume only Nclumps (∼ 20 to 50) to contain a signal given the NOEMA 2′′ × 2′′ synthesised beam. Since we will measure smaller regions of the two superimposed galaxies, we will also expect to probe only a fraction, σv, of the global velocity gradient. These local velocity dispersions are expected to be about 30km s−1 for the studied galaxy. We thus expect to find a signal strength of approximately F/(Nclumps × σv ) in each beam. In Tab. 5.2, we give the estimated signal in each beam for the proposed observations. We furthermore show the expected noise RMSexp computed with the IRAM sensitivity estimator. We also plan to search for the continuum flux (expected at 30μJy and 12μJy respectively in the 2′′ × 2′′ synthesised beam) which might be associated to the VLA flux. We finally find for a telescope time of 4h for each CO line an estimated signal strong enough to map the molecular gas with high spatial and spectral resolution. This project, 106
CHAPTER 5. EXPLORING THE COLD GAS CONTENT OF THE DP/MANGA GALAXIES 5 Observations at NRT 5.1 Data acquisition and reduction
We observed the 16 lowest-redshifted DP/MaNGA galaxies with the Nançay Radio Telescope (NRT), located in France. The radio telescope is composed of a rotating plane mirror that reflects the waves towards the spherical mirror which sends them to the focal carriage in the centre, where they are collected by cooled receivers. At 21 cm, the half power beam widths are 4′ (R.A.)× 22′ (Dec.). The radio telescope is composed of a plane mirror that reflects the waves Figure (5.8
) Left panel: Photography of the NRT. Right panel: Photography of the focal carriage and schematic representation of the wave path. towards the spherical mirror which sends them to the focal carriage in the centre, where they are collected by cooled receivers. We used the Low Frequency system which is optimised for the HI 21 cm line and recorded four polarisations simultaneously. Figure (5.9) Atomic gas spectra for the detected DP/MaNGA galaxies
.
5. OBSERVATIONS AT NRT 5.2 107 Atomic gas analysis
For all detected galaxies, we compute the atomic gas mass using: MHI M⊙ 2.356 × 105 = (1 + z) DL Mpc 2 FHI Jy km s−1 (5.12) where DL is the luminosity distance of the source and FHI is the velocity-integrated flux (Giovanelli & Haynes, 2016). We have started computing the HI masses and upper limits for the galaxies that we observed. We will be able to compare our results with the measurements from HI-MaNGA (Masters et al., 2019; Stark et al., 2021) and ALFALFA surveys, when available. 5.3 Perspectives Combining both the CO and the HI data will allow me to infer the cold gas content of 16 of the DP/MaNGA galaxies. I will be able to derive a Kennicutt-Schmidt relation based on both the molecular and atomic gas. 108
CHAPTER 5. EXPLORING THE COLD GAS CONTENT OF THE DP/MANGA GALAXIES
Conclusion Ce travail porte sur les mécanismes responsables de l’arrêt de la formation d’étoiles dans les galaxies proches, à travers l’étude de la morphologie, de la cinématique et de l’excitation du gaz ionisé. Afin de connaître le contenu en gaz froid, constituant le réservoir de formation d’étoiles, au sein de ces galaxies locales, la quantité de gaz moléculaire et atomique est également estimée. En utilisant le relevé Mapping Nearby Galaxies at APO (MaNGA), nous avons effectué une analyse des propriétés résolues spatialement d’un échantillon de 29 galaxies proches. Dans ce travail, nous nous concentrons sur l’étude d’objets qui présentent tous la même particularité : des caractéristiques spectrales complexes sont détectées dans leur spectre intégré dans les 3′′ centrales, qui sont bien ajustées par une approche d’ajustement à plusieurs composantes. Nous avons étudié différentes raies d’émission de gaz ionisé (notamment Hβ, [OIII], Hα, [NII]) afin d’étudier les processus de formation et d’extinction des étoiles. Nous avons inspecté certains indices spectraux, afin de déduire les propriétés des populations stellaires des 29 galaxies que nous avons analysées. Nous avons également commencé à étudier la cinématique du gaz et des étoiles au sein de ces galaxies, afin de sonder les mécanismes sous-jacents responsables des caractéristiques spectrales complexes que nous observons. Pour estimer l’efficacité de la formation d’étoiles, nous avons réalisé des observations du gaz moléculaire à l’IRAM 30 m et calculé la masse totale d’hydrogène moléculaire. Dans le but de sonder le contenu global en gaz froid de ces galaxies, nous avons également commencé à évaluer leur masse de gaz atomique, sur la base de nos observations au radio-télescope Nançay pour 16 galaxies. Les principaux résultats concernant ces 29 galaxies sont résumés ci-dessous: Grâce à l’analyse des données MaNGA, nous avons remarqué que les 29 galaxies DP/MaNGA montrent un excès de formation d’étoiles dans un rayon effectif. En étudiant les traceurs de l’âge des populations stellaires ainsi que la source de l’excitation du gaz ionisé, nous avons établi que ces galaxies constitu une séquence évolutive, compatible avec un scénario de fusions mineures. Les galaxies appartenant au «nuage bleu» sont des galaxies qui présentent une formation d’étoiles active et des populations stellaires jeunes à tous les rayons galactiques, et qui ont une grande quantité de gaz, probablement due à une accrétion de gaz. Les galaxies de la «vallée verte» que nous observons doivent leur couleur à des populations stellaires rouges et présentent deux types de distributions spatiales: certaines galaxies ont une excitation de type AGN dans leurs régions centrales, ainsi que des populations stellaires âgées. Elles pourraient correspondre à des galaxies dont l’activité nucléaire a été déclenchée par des flux de gaz vers le centre, éteignant la
CHAPTER 5. EXPLORING THE COLD GAS CONTENT OF THE DP/MANGA GALAXIES
formation d’étoiles “de l’intérieur vers l’extérieur”. D’autres galaxies ne montrent pas de claire évidence d’excitation type AGN en leur centre et possèdent de vieilles populations stellaires dans leurs régions périphériques. Nous affirmons que le gaz dans ces galaxies a été entraîné des parties extérieures vers le noyau et que l’épuisement du gaz extérieur entraîne une extinction de la formation d’étoiles “de l’extérieur vers l’intérieur”. Dans l’une de ces 29 galaxies, notre propre procédure d’ajustement, basée sur une approche à plusieurs composantes, a révélé la présence de deux objets sur la ligne de visée. Nous avons associé chaque composante cinématique à une galaxie individuelle et estimé un rapport de masse de 1:9 pour ce système en interaction. De plus, nous avons remarqué une émission radio étendue superposée au compagnon le plus petit, et avons détecté une métallicité du gaz élevée et une région de formation d’étoiles excentrée dans la galaxie principale. Ces caractéristiques peuvent résulter d’échanges de gaz pendant le processus de fusion et, afin de comparer la cinématique du gaz ionisé avec celle du gaz moléculaire, nous avons demandé des observations à haute résolution du gaz CO avec l’interféromètre NOrthern Extended Millimetre Array. La première inspection des données indique une distribution irrégulière du gaz moléculaire. L’analyse effectuée sur cette galaxie s’est avérée efficace pour dévoiler les mécanismes sousjacents responsables des caractéristiques spectrales que nous avons détectées. Par la suite, nous utiliserons donc cette analyse à plusieurs composantes pour étudier et interpréter la cinématique des autres galaxies à doubles-pics/MaNGA. Des analyses innovantes telles que celle que nous avons appliquée aideront à obtenir une compréhension satisfaisante des processus qui déclenchent, régulent et éteignent la formation d’étoiles, et par conséquent, à résoudre certaines des questions restantes sur l’évolution des galaxies.
6 Conclusion
This work addresses the mechanisms responsible for star formation quenching in nearby galaxies, through the study of the morphology, the kinematics and the excitation of the ionised gas. In order to inquire the cold gas content, constituting the star formation reservoir, within these local galaxies, the amount of molecular and gas is also estimated. Using the Mapping Nearby Galaxies at APO –MaNGA– survey, we have performed an analysis of the spatially-resolved properties of a sample of 29 nearby galaxies. In this work, we focus on the study of objects that all show the same peculiarity: they present complex spectral features in their central 3′′ spectrum, which are well recovered by a multi-component fitting approach. We investigated different ionised-gas emision lines (notably Hβ, [OIII], Hα, [NII]) to study star formation and termination processes. We inspected some spectral indices, in order to infer stellar population properties of the 29 galaxies that we analysed. We have also started to investigate the kinematics of gas and stars within these galaxies, so as to probe the underlying mechanisms reponsible for the complex spectral features that we observe. To estimate the star formation efficiency, we have performed molecular gas observations at IRAM 30 m and computed the total mass of molecular hydrogen. Aiming at probing the global cold gas content of these galaxies, we have started evaluating their atomic gas mass as well, based on our observations at the Nançay Radio Telescope for 16 galaxies. The main conclusions about these 29 galaxies are summarised below: Through the analysis of the spatially-resolved data, we showed that the star formation excess, in these 29 galaxies, is located within one effective radius. By classifying them according to the AGN activity evidence, as well as their stellar population age, we established that these galaxies constitute an evolutionary sequence, compatible with a minor merger scenario. The blue group gathers galaxies that exhibit star formation and young stellar populations at all radii, that have a large amount of gas possibly due to gas accretion. The green objects that we observe owe their colour to red stellar populations and present two types of spatial distributions. Some galaxies have an important AGN contribution in their inner parts, which correlates with a lack of young stars. They might correspond to galaxies whose nuclear activity has been triggered by gas fueling towards the centre, quenching star formation “inside-out”. Some others do not show any strong AGN evidence at their centre and host old stellar populations in their outskirts. We In one of these 29 galaxies, our own fitting procedure, based on a multi-component approach, revealed the presence of two objects in the line of sight. We associated each kinematics component to an individual galaxy and estimated a mass ratio of 1:9 for this interacting system. Furthermore, we noticed an extended radio emission superimposed on the smaller companion, and detected off-centered high gas-phase metallicity and star formation in the main component. These characteristics can result from gas exchanges during the merging process and so as to compare the ionised gas kinematics with that of the molecular gas, we applied for high-resolution CO gas observations with the NOrthern Extended Millimetre Array. The first inspection of the data indicates an irregular distribution of the molecular gas. The analysis performed on this galaxy proved to be efficient at unveiling the underlying mechanisms responsible for the spectral features that we detected. Next, we will thus make use of this multi-component analysis to study and interpret the kinematics of the other double-peaked/MaNGA galaxies. Innovative analyses such as the one that we applied will help obtain a satisfactory understanding of the processes that trigger, regulate and quench star formation, and consequently, solve some of the remaining questions about galaxy evolution.
7 Appendices 1 Workshop and conference participations
1.1 Workshops and conferences •
February
2021 (
online
)
Double-peak emission line galaxies workshop, organising: committee member “Morphokinematic transformations in an interacting local galaxy in the light of MaNGA”, oral presentation • 2019
&
2020 (online) European Astronomical Society Annual Meeting Lyon: “Morpho-kinematic transformation observed in double-peak emission-line MaNGA galaxies”, poster presentation • May 2019 (Québec, Canada) SIGNALS survey workshop: “Asymmetric kinematics features within galaxies at z=0.06 as revealed by MaNGA”, oral presentation. • October 2019 (Paris)
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Design and Experimental Study of Progressive Spring for Formula Student Race Car
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Anmol Shripad Patil
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1. INTRODUCTION: have a constant rate of deflection. For this reason,
they are called variable rate springs. Since racetracks
have different road surfaces, a suspension that is
adaptive to changing with road surfaces is desired. It
also means to achieve a compliant suspension in the
rough and a tight suspension for high-speed turns. Whereas linear springs allow working easily because
the spring rate is constant, this results in quick tuning
as many teams avoid risking in terms of suspension
settings. Hence progressive springs become a great
advantage. have a constant rate of deflection. For this reason,
they are called variable rate springs. Since racetracks
have different road surfaces, a suspension that is
adaptive to changing with road surfaces is desired. It
also means to achieve a compliant suspension in the
rough and a tight suspension for high-speed turns. Whereas linear springs allow working easily because
the spring rate is constant, this results in quick tuning
as many teams avoid risking in terms of suspension
settings. Hence progressive springs become a great
advantage. FS car’s suspension springs compress to absorb
impact when tires roll over irregular terrain and keep
the car off the ground in correspondence with the
damper. The two types of springs used in race cars
are linear and progressive. Which one do you think
is best for FS cars? On the one hand, progressive-rate
springs generally have less total deflection; they also
provide the benefits of a lowered car - better
handling, reduced nose dive, and decreased body
roll, along with better ride quality. This is because
the closely wound coils needed to create the dual
rates take up deflection space. As a progressive
spring compresses, the spring rate will gradually
increase. On the other hand, in linear springs, the
spring rate is constant, and it is not affected by the
load acting on the spring. In this paper, a detailed
design procedure and analytical calculation of
progressive spring for FS car followed by the
experimental study whose results were obtained by
spring load testing machine are explained. But even after choosing progressive, how will one
assure whether it is satisfactory or not? Rather how
will one determine the behavior of spring? So that is
when the experimental study comes into play which
has to go hand in hand with the theoretical research. 2. METHODOLOGY: 1. To achieve better ride quality and superior
handling due to the roll control offered by the
springs. Original Article License: This work is licensed under a Creative Commons Attribution 4.0 International License. R
d F ll Li 1. INTRODUCTION: The load testing setup provides us with estimated
results that are further validated with results from
testing a car and running it on progressive springs. And then, it leads to the final lock to the tuning. Abstract: The purpose of carrying out the present work is to design, manufacture & test the progressive springs on an FS
vehicle. This is one type of helical spring with a variable spring rate. The main purpose of designing progressive
springs is to avail all the advantages of the variable spring rate over the linear spring rate and better ride quality
along with roll control, compared to linear rate springs. We took several factors of vehicle dynamics under
consideration before settling on progressive springs. Before starting with the design procedure, we had set
objectives and followed the standard methodology of spring design to get the required output. The purpose of carrying out the present work is to design, manufacture & test the progressive springs on an FS
vehicle. This is one type of helical spring with a variable spring rate. The main purpose of designing progressive
springs is to avail all the advantages of the variable spring rate over the linear spring rate and better ride quality
along with roll control, compared to linear rate springs. We took several factors of vehicle dynamics under
consideration before settling on progressive springs. Before starting with the design procedure, we had set
objectives and followed the standard methodology of spring design to get the required output. Along with that, we took design philosophy under consideration. We reviewed all the parameters before
finalizing the spring material as it is one of the major factors. We carried out all the necessary design calculations
to complete the dimensions and stiffness of the spring. The conclusion helped us to achieve better ride quality
and roll control accompanying the optimized spring design satisfying all the necessities such as load, stiffness,
and deflection of progressive springs. Along with that, we took design philosophy under consideration. We reviewed all the parameters before
finalizing the spring material as it is one of the major factors. We carried out all the necessary design calculations
to complete the dimensions and stiffness of the spring. The conclusion helped us to achieve better ride quality
and roll control accompanying the optimized spring design satisfying all the necessities such as load, stiffness,
and deflection of progressive springs. Keywords: suspension; progressive springs; spring rate; ride quality; compression Anmol Shripad Patil1, Eshita Nandi2, Prasad Nanasaheb Punekar3* ,Suyash Wagh4 *Correspondence (Prasad Nanasaheb Punekar): prasadpunekar29@gmail.com
1 Flat no. D-303, Olympia Society, Wakad, Pune-411056. 2 Flat no.403 ,A2 building,Manik Baug Orchid,opp anna magar saheb stadium,pimpri pune -18
3 Abhang Colony, Rahatani, Kalewadi, Pune-411017. 4 Building No.16, Room No. 06, Police Headquarters, Gangapur Road, Nashik-4220 *Correspondence (Prasad Nanasaheb Punekar): prasadpunekar29@gmail.com
1 Flat no. D-303, Olympia Society, Wakad, Pune-411056. 2 Flat no.403 ,A2 building,Manik Baug Orchid,opp anna magar saheb stadium,pimpri pune -18
3 Abhang Colony, Rahatani, Kalewadi, Pune-411017. 4 Building No.16, Room No. 06, Police Headquarters, Gangapur Road, Nashik-4220 2.3 Abbreviations Fig. 1 Flow chart of a design procedure for springs Fig. 1 Flow chart of a design procedure for springs 2.1 Problem Statement 𝐿𝑓1- free length of softer region of spring
16. 𝐿𝑓2- free length of stiffer region of spring
17. D - mean diameter of coil
18. G / g - Modulus of Rigidity
19. n1 - total number of coils
20. n - total active number of coils
21. n1 -total active number of coils of softer region of
spring
22. n2 -total active number of coils od stiffer region of
spring
2.4 Design Procedure
Fig. 1 Flow chart of a design procedure for springs 10. 𝛿1- deflection of softer region of spring
11. 𝛿2- deflection of stiffer region of spring
12. d - spring diameter
13. 𝐿𝑆- solid length of spring
14. 𝐿𝑓- free length of total number of coils
15. 𝐿𝑓1- free length of softer region of spring
16. 𝐿𝑓2- free length of stiffer region of spring
17. D - mean diameter of coil
18. G / g - Modulus of Rigidity
19. n1 - total number of coils
20. n - total active number of coils
21. n1 -total active number of coils of softer region of
spring
22. n2 -total active number of coils od stiffer region of
spring 2.1 Problem Statement 2. To get a lower ride height, increased stiffness
through hard cornering, and a more comfortable ride
over a Why choose progressive rate springs over linear rate
springs? Progressive springs contribute to the roll
control of the vehicle. A progressive spring does not 1 stiffer ride. 3. To avail the advantages of the dual stiffness over
the different track conditions. 4. To validate the design by correlating design
calculations with the testing results data. 2.3 Abbreviations
1. FS- formula student
2. k - spring stiffness
3. k1- spring stiffness of softer region of coil
4. k2 - spring stiffness of stiffer region of coil
5. W - load acting on spring
6. W1 - load acting on softer region of coil
7. W2 - load acting on stiffer region of coil
8. 𝛿- deflection of spring
9. 𝛿𝑚𝑎𝑥- maximum deflection of spring
10. 𝛿1- deflection of softer region of spring
11. 𝛿2- deflection of stiffer region of spring
12. d - spring diameter
13. 𝐿𝑆- solid length of spring
14. 𝐿𝑓- free length of total number of coils
15. 𝐿𝑓1- free length of softer region of spring
16. 𝐿𝑓2- free length of stiffer region of spring
17. D - mean diameter of coil
18. G / g - Modulus of Rigidity
19. n1 - total number of coils
20. n - total active number of coils
21. n1 -total active number of coils of softer region of
spring
22. n2 -total active number of coils od stiffer region of
spring
2.4 Design Procedure
Fig. 1 Flow chart of a design procedure for springs stiffer ride. 3. To avail the advantages of the dual stiffness over
the different track conditions. 4. To validate the design by correlating design
calculations with the testing results data. 2.3 Abbreviations
1. FS- formula student
2. k - spring stiffness
3. k1- spring stiffness of softer region of coil
4. k2 - spring stiffness of stiffer region of coil
5. W - load acting on spring
6. W1 - load acting on softer region of coil
7. W2 - load acting on stiffer region of coil
8. 𝛿- deflection of spring
9. 𝛿𝑚𝑎𝑥- maximum deflection of spring
10. 𝛿1- deflection of softer region of spring
11. 𝛿2- deflection of stiffer region of spring
12. d - spring diameter
13. 𝐿𝑆- solid length of spring
14. 𝐿𝑓- free length of total number of coils
15. 2.5 Design Philosophy 1. Calculation of force to be applied on spring. 2. Figuring out solid height will also be based on the
calculated force. The spring should not compress to
its solid height before all the force is applied. 3. Based on this calculated force, wire is selected. 4. Spring design takes careful consideration, precise
calculations, and the right expertise. 5. Spring model. The selection of spring materials depends on the
following factors: 1. Calculation of force to be applied on spring. The selection of spring materials depends on the
following factors: 2. Figuring out solid height will also be based on the
calculated force. The spring should not compress to
its solid height before all the force is applied. g
1. Maximum load acting on the spring. 2. Set of stress through which the spring operates g
pp
3. Based on this calculated force, wire is selected 3. Constraints of mass and volume of spring. 4. Expected fatigue life. 4. Spring design takes careful consideration, precise
calculations, and the right expertise. 5. Environmental conditions such as temperature
and corrosive atmosphere. 5. Spring model. 6. Severity of deformation encountered while
making spring. 2 2.7 Spring Parameters Sr. No. Parameter
Specification
1
Raw Material
Sp.St.GR-III
2
Wire Diameter
6.00 mm
3
Outside Diameter
47.00 mm
4
Mean Diameter
41.00 mm
5
Total Coil
12.00 Nos
6
Active Coil
10.00 Nos
7
Free Length
140.00 mm
8
Coil Upper Length (Low Pitch)
65.00 mm
9
Coil Lower Length (High Pitch)
75.00 mm
10
Solid Length
72.00 mm
Table 1: Design parameters and respective specifications of progressive spring. Table 1: Design parameters and respective specifications of progressive spring. Therefore n1 =
𝑔𝑑 4
8𝐷3 𝑘1 =
79.344 ×103× 64
8×41.63×29
n1=6.1567 Therefore n1 =
𝑔𝑑 4
8𝐷3 𝑘1 =
79.344 ×103× 64
8×41.63×29 2.8 Design Calculation NOTE: a) Initially K is assumed on the basis of
maximum load acting, deflection required and free
length 𝐿𝑓1 = 𝐿𝑆+ 𝛿𝑚𝑎𝑥+ (𝑛1 −1) × 1 (2)
𝐿𝑓1 = (𝑛1𝑑) + 𝛿𝑚𝑎𝑥+ (𝑛1 −1) × 1 (3)
𝑛1 = 𝑡𝑜𝑡𝑎𝑙 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑐𝑜𝑖𝑙𝑠. (2)
(3) b) If we need to increase the load capacity, we need
to increase the stiffness and decrease the deflection ↑𝑘= 𝑊
𝛿↓
d On each side of coil, the no. of inactive coil will be On each side of coil, the no. of inactive coil will be
1. 𝛿↓
c) To decrease the 𝐿𝑓, we need to decrease the
number of coils, i.e., increase the pitch Keeping
the stiffness constant. Therefore, Therefore, Therefore,
𝐿𝑓= (𝑛+ 1)𝑑+ 𝛿𝑚𝑎𝑥+ (𝑛1 + 1 −1) × 1 (4)
𝐿𝑓= (𝑛+ 1)𝑑+ 𝛿𝑚𝑎𝑥+ 𝑛× 1
𝐿𝑓1 = (n1 + 1) d +𝛿1+ n1×1
𝐿𝑓1 = (6.1567 + 1)6 + 𝛿1 + 6.1567 (5)
Constraint of deflection. (4) a)
d= 6 mm
D= 41.6 mm
k1= 29 N/mm (assumed stiffness)
G= 79.344× 103 N/mm^2
n1=? 𝐿𝑓1=? k=
𝒈𝒅 𝟒
𝟖𝑫𝟑𝒏 (1 a)
d= 6 mm
D= 41.6 mm
k1= 29 N/mm (assumed stiffness)
G= 79.344× 103 N/mm^2
n1=? 𝐿𝑓1=? k=
𝒈𝒅 𝟒
𝟖𝑫𝟑𝒏 (1) a)
d= 6 mm
D= 41.6 mm
k1= 29 N/mm (assumed stiffness)
G= 79.344× 103 N/mm^2
n1=? 𝐿𝑓1=? k=
𝒈𝒅 𝟒
𝟖𝑫𝟑𝒏 (5) 𝑓
Constraint of deflection. Constraint of deflection. Therefore,
Max deflection=𝛿𝑚𝑎𝑥= 55.8 mm -------- Total
But, we need, 𝑡𝑜𝑡𝑎𝑙 𝑑𝑒𝑓𝑙𝑒𝑐𝑡𝑖𝑜𝑛×
1
3-------(for low
pitch coils) Therefore, Therefore, Max deflection=𝛿𝑚𝑎𝑥= 55.8 mm -------- Total
But, we need, 𝑡𝑜𝑡𝑎𝑙 𝑑𝑒𝑓𝑙𝑒𝑐𝑡𝑖𝑜𝑛×
1
3-------(for low
pitch coils) (1) 3 𝛿1 = 55.8 ×
1
3
𝛿1 = 18.6
(6)
Now,
𝛿1 =
8𝑊𝐷3𝑛
𝐺𝑑 4 (7)
𝛿1=18.6 mm
D=41 6 mm
W = Total load that the coils can take
W =𝑊1 + 𝑊2 = 1915.7913 𝑁
Total coils = 4.8256+6.1567+Inactive coils
=10.9823+2
=12.9823
𝐿𝑓= 𝐿𝑓1 + 𝐿𝑓2=144.6758 mm W = Total load that the coils can take
W =𝑊1 + 𝑊2 = 1915.7913 𝑁
Total coils = 4.8256+6.1567+Inactive coils
=10.9823+2
=12.9823 (6) 𝐿𝑓= 𝐿𝑓1 + 𝐿𝑓2=144.6758 mm (7) (7) From (5) and (6) From (5) and (6)
𝐿𝑓1 = (6.1567 + 1)6 + 18.6 + 6.1567
𝐿𝑓1 = 67.6969 𝑚𝑚
𝑘=
𝑊
𝛿 equation satisfied (8) 𝐿𝑓1 = (6.1567 + 1)6 + 18.6 + 6.1567 (8) b) d= 6 mm
D= 41.6 mm
k2= 37 N/mm (assumed stiffness)
G= 79.344× 103 N/mm
n2=? 𝐿𝑓2= ? k2=
𝑔𝑑 4
8𝐷3𝑛 (1)
Therefore
n2 =
𝑔𝑑 4
8𝐷3𝑘2 =
79.344 ×103× 64
8×41.63×37
n2=4.8256
Remaining deflection= (55.8-18.6)
𝛿2=37.2 mm 3. RESULTS: The designed spring is being tested on an industrial
spring testing machine. The results of testing are
given in the form of load v/s deflection graph shown
in fig.2. Fig.2 Load v/s Displacement graph Fig.2 Load v/s Displacement graph From (5) and (6) Authors’ Information: Anmol Shripad Patil, born in 2001, is currently pursuing B.E. in
mechanical engineering with Savitribai Phule Pune University, Pune. He is interested in field of research on robotics and automation. Eshita Nandi, born in 1999, has completed B.E. in mechanical
engineering from Savitribai Phule Pune University, Pune. Her main
interest lies in research of automobiles. Prasad Nanasaheb Punekar, born in 2000, is currently pursuing B.E. in mechanical engineering with Savitribai Phule Pune University,
Pune. He is interested in field of research on mechanical design and
automation. Fig. 5 Design Model of Progressive Springs Suaysh Wagh, born in 1998, is pursuing M.Sc. mobility engineering
with Charmers University of Technology, Sweden. His is mainly
interested in automotives research. 4. DISCUSSION: The spring’s progressive behavior could be
observed from fig.2 Load v/s deflection graph. Initially, the graph is non-linear and then it turns into
linear which indicates that at the beginning the load
required to deflect the spring up to certain extent is
less, and afterward as the deflection goes on
increasing the load required increases linearly. (1) Fig. 3 Progressive springs on testing setup (2)
(3)
(4)
(5)
(6)
Fig. 3 Progressive springs on testing setup The maximum load is given by 𝛿2 =
8𝑊𝐷3𝑛2
𝐺𝑑 4 (2)
𝑊=
𝛿2×𝐺𝑑 4
8𝐷3𝑛2 (3)
W2=1376.3875 N Fig. 3 Progressive springs on testing setup Fig. 3 Progressive springs on testing setup 5 5 Fig. 4 Manufactured and tested model
Fig. 5 Design Model of Progressive Springs Fig. 4 Manufactured and tested model Fig. 4 Manufactured and tested model length, maximum bump travel, and droop travel. The iteration we performed to achieve the required
stiffness within the constraints was successful, and
the design gave us the needed results on testing. We
validated the spring design with the help of actual
testing. Authors’ Contributions: Fig. 4 Manufactured and tested model Anmol Shripad Patil was in charge of the complete research and
design process. Prasad Nanasaheb Punekar and Suyash Wagh
together monitored complete experimentation and manufacturing
process. Eshita Nandi managed and analyzed the testing. Fig. 5 Design Model of Progressive Springs Acknowledgement: Acknowledgement:
We thankfully acknowledge Suaan Techmechs Pvt Ltd of India and
Team Redline Racing of JSPM’s Rajarshi Shahu College of Engineering
of India for their support. Funding: Competing Interests:
No competing financial and non-financial interests. Availability of Data and Materials:
Not Applicable. Funding:
Not Applicable. 5. CONCLUSIONS: In this research paper, part of the discussion is a
study about the design of progressive springs for FS
cars. This attempt was helpful to get successful
results, as we tested the progressive springs on the
FS car and an industrial spring testing machine. The
stiffness of 29 N/mm at the softer region and 37
N/mm at the stiffer region was being finalized, and
the spring with this stiffness fulfilled the aim of both
the softer rate at the low load and the stiffer rate for
the sharp turns. The stiffness of springs increased
with increasing load in parabolic nature. This
fascinated the required load transfer at corners. Its
starting low stiffness also provided comfort to the
driver at small bumps and droops. It provides the
required stiffness as well as the maximum strength,
which was necessary for our design. At the same
time, it also satisfied all the constraints of free REFERENCES: 1. https://www.hypercoils.com › linear...Web results Linear vs. Progressive
Rate Suspension Springs | Tech Tips | Hyperco
2. Design Of Machine, McGraw hill, VB Bhandari
3. https://www.progressivesuspension.com
4. Web results
5. Progressive Suspension: High-performance motorcycle suspension ... 6. https://en.m.wikipedia.org › wiki
7. Spring (device) – Wikipedia
8. RCVD (Race Car Vehicle Dynamics) 5 5
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Exposure to Celebrity-Endorsed Small Cigar Promotions and Susceptibility to Use among Young Adult Cigarette Smokers
|
Kymberle L. Sterling
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Hindawi Publishing Corporation
Journal of Environmental and Public Health
Volume 2013, Article ID 520286, 6 pages
http://dx.doi.org/10.1155/2013/520286 Hindawi Publishing Corporation
Journal of Environmental and Public Health
Volume 2013, Article ID 520286, 6 pages
http://dx.doi.org/10.1155/2013/520286 Kymberle L. Sterling,1 Roland S. Moore,2 Nicole Pitts,1 Melissa Duong,1
Kentya H. Ford,3 and Michael P. Eriksen1 1 Institute of Public Health, Georgia State University, 140 Decatur Street, Urban Life Building, Room 878, Atlanta, GA 30303, USA
2 Prevention Research Center, Pacific Institute for Research and Evaluation, 1995 University Avenue,
SteX 450, Berkeley, CA 94704, USA 3 Health Outcomes and Pharmacy Practice Division, The University of Texas College of Pharmacy,
2409 University Avenue PHR Building, 3.209 A 1900, Austin, TX 78712-0127, USA 3 Health Outcomes and Pharmacy Practice Division, The University of Texas College of Pharmacy,
2409 University Avenue PHR Building, 3.209 A 1900, Austin, TX 78712-0127, USA Correspondence should be addressed to Kymberle L. Sterling; ksterling@gsu.edu Correspondence should be addressed to Kymberle L. Sterling; ksterling@gsu.edu Received 2 July 2013; Revised 25 September 2013; Accepted 8 October 2013 Received 2 July 2013; Revised 25 September 2013; Accepted 8 October 2013 Academic Editor: Ike S. Okosun Copyright © 2013 Kymberle L. Sterling et al. This is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly
cited. Small cigar smoking among young adult cigarette smokers may be attributed to their exposure to its advertisements and promotions. We examined the association between exposure to a celebrity musicartist’s endorsement of a specific brand of small cigars and young
adult cigarette smokers’ susceptibility to smoking that brand. Venue-based sampling procedures were used to select and survey a
random sample of 121 young adult cigarette smokers, aged 18–35. Fourteen percent reported exposure to the artist’s endorsement of
the small cigar and 45.4% reported an intention to smoke the product in the future. The odds of small cigar smoking susceptibility
increased threefold for those who reported exposure to the endorsement compared to those not exposed (OR = 3.64, 95% CI 1.06
to 12.54). Past 30-day small cigar use (OR = 3.30, 95% CI 1.24 to 8.74) and past 30-day cigar use (OR = 5.08, 95% CI 1.23, 21.08) were
also associated with susceptibility to smoke a small cigar. An association between young adult cigarette smokers’ exposure to the
music artist’s small cigar endorsement and their susceptibility to smoke small cigars was found. This association underscores the
importance of monitoring small cigar promotions geared toward young people and their impact on small cigar product smoking. Journal of Environmental and Public Health Journal of Environmental and Public Health traditional avenues for cigar advertisements, tougher restric-
tions on tobacco advertising may entice the tobacco industry
to use creative marketing strategies to promote small cigars. According to a small survey of concertgoers (𝑛= 238) posted
on the festival’s webpage (http://www.coachella.com), 31.5%
were aged 25–29 years, 29.8% were aged 20–24 years, and
12.2% were aged 30–34 years old. Venue-based sampling pro-
cedures [23], in which the venue is the primary sampling unit,
were used to select and survey eligible young adults at the
festival. to use creative marketing strategies to promote small cigars. In 2012, a celebrity hip-hop music artist, Calvin Broadus
Jr., also known as Snoop Dogg, announced his launching
of a new brand line of cigarillo products, called Executive
Branch. Several widely circulated music magazines and press
releases reported that Snoop Dogg planned to “unveil” the
new product during his upcoming performances at the
2012 Coachella Music and Arts Festival in Indio, California. The music festival mainly attracts an international audience
of adults aged 35 and younger [10, 11]. A recent study
by Richardson and colleagues recently documented Snoop
Dogg’s promotion of Executive Branch small cigars on social
media sites, such as Instagram [12]. The tobacco industry
historically has relied upon hip-hop and other forms of music
to promote smoking, as music can be used as a form of direct,
targeted marketing [13]. Hip-hop music has a wide appeal
and a varied fan base across different genders, racial/ethnic
backgrounds, and nationalities (i.e., USA versus international
countries). Endorsements by its celebrity music artists can
introduce their youthful fan base (primarily under 30 years
old and including many fans under the age of 18) to new
tobacco products and smoking behavior. As such, exposure
to a hip-hop music artist’s endorsement and promotion of
a brand-specific small cigar product may be associated with
young adults’ small cigar smoking behaviors. In 2012, the
Surgeon General’s report concluded that there is a causal
association between exposure to smoking in the movies and
youth smoking. The report also concludes that traditional
advertising causes youth smoking [14, 15]. To our knowledge
no study has examined the association between small cigar
smoking behavior and exposure to a celebrity music artist’s
advertisement and promotion of small cigars. Journal of Environmental and Public Health We surveyed
a random sample of young adult cigarette smokers who
attended the Coachella festival in Indio, California, to assess
their exposure to Snoop Dogg’s endorsement and advertise-
ment for Executive Branch small cigars. We focused on young
adult cigarette smokers because extant evidence suggests that
they may be considered at high risk for small cigar use
[6, 7, 16]. We hypothesized that young adults who reported
exposure to the artist’s small cigar advertisements, compared
to those who did not report exposure, would have greater
susceptibility or intention to smoke Executive Branch small
cigars in the future. Our hypothesis is supported by ample
evidence that has reported that exposure to cigarette smoking
advertisements is associated with a susceptibility to smoke
cigarettes [17–20]. Our rationale for use of intention to smoke
small cigars is that it is a proximal predictor of smoking
behavior [21] and is the most appropriate outcome to assess
i
ti
l d i To construct our sampling frame, the study team (K.S. and N.P.) utilized ethnographic sampling techniques to sur-
vey the area and identify locations within the festival where
young adult smokers would gather or pass through. We
identified three locations within the venue that fit this crite-
rion: two “beer gardens” where festival attendees purchased
and consumed alcoholic beverages and a smoking lounge. Each location was restricted to individuals aged 18 and older. Within the smoking lounge, we implemented area-based
sampling and approached each individual for inclusion into
the survey. For the two “beer gardens,” we identified young
adults who were smoking cigarettes and randomly selected
them by systematically intercepting every third cigarette
smoking attendee that crossed a predetermined point. 2.2. Procedures. The two study team members collected data
from attendees over a two-day period from festival open to
close. Upon interception, each team member approached fes-
tival attendees, gave them a short description about the study,
and obtained verbal consent. If the attendee refused, the study
team member noted their refusal and continued the process
with the next random participant. Festival attendees who
appeared to be visibly impaired by alcohol or other substances
were not approached. The study team determined the eligibil-
ity status of festival attendees that were intercepted and who
were willing to participate using a brief screener that verified
the attendant’s age and cigarette smoking status. Attendees
who did not meet the eligibility criteria were not surveyed. Journal of Environmental and Public Health g
y
y
Over the two-day data collection period, a total of 275
participants were approached. Of those 126 (45.8%) refusals
were noted and 6 (2.2%) participants did not meet study
eligibility criteria. Data were lost for 22 respondents due to a
synchronization error between the server and the device and
could not be retrieved. Thus survey responses were collected
and retained for 121 young adults. 2.3. Study Sample Description. The majority of the 121
respondents were male (55.4%) and between the ages of 18
and 30 years old (88.0%). Regarding race/ethnicity, most
respondents were white (75.2%) and lived in a country
outside of the USA (52.4%). Overall, 70% reported past 30-
day nonmentholated cigarette smoking, while 36.4% smoked
menthol cigarettes. 2.4. Measures. Survey items were entered in a survey software
app, iSurveySoft (http://www.isurveysoft.com), and were
uploaded to the handheld device (iPod touch 5th generation). The brief survey consisted of 21 items including demographic
information, smoking history and current tobacco use, and
exposure to small cigar advertisements. 1. Introduction sustain dependence and increase the carbon monoxide levels
of smokers [3, 4]. Small cigar smoking is a public health threat
and is not a safer alternative to cigarette smoking. In the United States (USA), cigarette consumption has
steadily declined during the past decade. However, sales of
cigars, particularly little cigars and cigarillos, have markedly
increased. Of the three types of cigar products available in
the USA, trend data from 1993 to 2006 suggest that the
sales of large cigars decreased from 37% to 47%, while the
sales of cigarillos increased from 25% to 32% and little cigars
increased from 37% to 47% [1]. Increased consumption of
small cigars (i.e., little cigars and cigarillos) presents a new
challenge for tobacco control researchers. Small cigars are
similar to cigarettes in their size, shape, and filtering, are often
inhaled, and are flavored. All cigars contain approximately the
same toxic and carcinogenic compounds found in cigarettes
[2]. Small cigars deliver considerable amounts of nicotine to g
g
Small cigar smoking has become increasingly popular
among young adults, aged 18–35. Over 65% of young adults
have heard about small cigars [5], and 26% report ever use of
small cigars [6]. Young adults with a history of tobacco use
have an elevated risk for cigar smoking, with recent studies
indicating that 12.0–16.0% of young adult cigarette smokers
have smoked some cigar variant (cigars, little cigar, or
cigarillos) in the past 30 days [6–8]. The increased popularity
of small cigars among young adult smokers may be due, in
part, to their exposure to advertisements and promotions of
small cigar products. Although magazines and point-of-sale
advertisements (which accounted for 24.1% and 13.0% of cigar
advertising and promotional expenditures in 1999) [9] are 2 2 3. Results 3.1. Small Cigar Smoking Behavior and Demographic Charac-
teristics. Of the 121 young adult cigarette smokers in our sam-
ple, 25.6% reported smoking small cigars at least once in the
past 30 days. Table 1 shows the distribution of demographic
characteristics by small cigar smoking status. The majority of
small cigar smokers in our sample were male and aged 18–
24 years old, though a sizeable proportion were aged 25–30
years old. Although most small cigar smokers in our sample
were White, significantly more African-Americans reported
smoking a small cigar in the past 30 days than not smoking a
small cigar (𝑃= 0.001). Notably, over half of the small cigar
smokers in our sample lived in a country outside of the United
States. 2.4.3. Susceptibility to Smoke Small Cigars in the Future. Respondents were asked two questions: (1) if they intended to
smoke Executive Branch small cigars and (2) if they intended
to smoke any other small cigar in the future. Response
categories for both questions were “yes” or “no.’’ 3.2. Concurrent Tobacco Use among Small Cigar Smokers. Small cigar smokers (𝑁
=
31) in our sample reported
concomitant use of other tobacco products. Over two-thirds
(71.0%) of small cigar smokers reported smoking nonmen-
tholated cigarettes in the past 30 days, while one-third (32.3%)
reported using mentholated cigarettes. Small cigar smokers
also reported current cigar smoking, with 19.4% reporting
smoking cigars at least once in the past 30 days. 2.4.4. Exposure to Small Cigar Advertisements. We assessed
festival attendees’ exposure to (1) the Snoop Dogg’s adver-
tisements of the Executive Branch small cigars and (2) other
advertisements for other small cigar products (e.g., Swisher
Sweets). Brand names were provided to assist the respondents
with recall. Response categories for both questions were
“yes” or “no.” If a respondent reported exposure to the
product, they were asked to recall where they had seen the
advertisement. Response categories included in a newspaper,
in a magazine, on the Internet, on social media sites (e.g.,
Facebook, Twitter, etc.), and other. 3.3. Exposure to Small Cigar Advertisements and Urge to
Smoke. Approximately 14.0% of respondents reported see-
ing Snoop Dogg’s advertisement of Executive Branch small
cigars. The majority of respondents (33.3%) reportedly saw
these advertisements on the Internet, while 27.8% reported
seeing them on social media sites, such as Facebook or
Instagram. 2. Methods 2.1. Participants and Sampling Frame Construction. Eligible
participants were young adults, aged 18–35, who attended the
Coachella festival and reported smoking at least one cigarette
in the past 30 days. Approximately 85,000 concertgoers
attended the festival per day over a three-day period [22]. 2.4.1. Demographic Information. Respondents were asked
about their age, gender, race/ethnicity, and country and 3 Journal of Environmental and Public Health state of residence. Age categories included <18 years old,
18–24 years old, 25–30 years old, and 31 years or older. Racial/ethnic categories included Asian/Pacific Islander,
American Indian/Alaskan Native, Black/African-American,
Hispanic, White, and other. Respondents were allowed to
select one or more of the racial/ethnic categories. who did not report use. Descriptive statistics were also used
to assess exposure to small cigar advertisements (celebrity-
endorsed and other) and the urge to smoke small cigars after
viewing the ads. Bivariate and multivariate logistic regression
analyses were conducted to assess the associations between
small cigar smokers and nonusers and the demographic,
smoking-related, exposure, and urge to smoke variables. The
dependent variable for the bivariate and multivariate analysis
was small cigar smoking. 2.4.2. Smoking-Related Variables. Respondents were asked
if they had used any of the following tobacco products in
the past 30 days: cigarettes, nonmentholated (i.e., Marlboro),
mentholated cigarettes (i.e., Newports), cigars, and small
cigars (e.g., Black & Mild, Swisher Sweets). The response
categories for each included “yes” or “no”. Brand names
were provided for each tobacco product to help respondents
better recognize the tobacco product [24]. Respondents were
allowed to select one or more of the tobacco products if they
used them in the past 30 days. Those who selected “yes”
for the past 30-day use of any of the tobacco products were
classified as current users. Journal of Environmental and Public Health the demographic and smoking-related variables and the
exposure to Snoop Dogg’s Executive Branch small cigar
advertisements were associated with susceptibility to smoke
the product in the future. Though demographic variables
were not associated with susceptibility, bivariate analyses
found that past 30-day cigar (OR = 4.90, 95% CI 1.26, 19.13,
𝑃= 0.02) and small cigar (OR = 2.98, 95% CI 1.19, 7.42,
𝑃= 0.02) smoking were significantly associated with an
intention to smoke Executive Branch small cigars. Exposure
to Snoop Dogg’s advertisements was marginally associated
with intention to smoke Executive Branch small cigars (𝑃=
0.08). Given the exploratory nature of our study, variables
from the bivariate analysis that were significant at 𝑃< 0.10
were included in the multivariate analysis. Table 2 shows the
results of our multivariate model. Respondents who reported
current cigar smoking were five times more likely and those
who smoked small cigars were three times more likely to
be susceptible to smoking Executive Branch small cigars. Respondents who reported seeing the artist’s advertisement
for Executive Branch small cigars were three times more likely
to be susceptible to smoking those small cigars than those
who were not exposed to the advertisement. the demographic and smoking-related variables and the
exposure to Snoop Dogg’s Executive Branch small cigar
advertisements were associated with susceptibility to smoke
the product in the future. Though demographic variables
were not associated with susceptibility, bivariate analyses
found that past 30-day cigar (OR = 4.90, 95% CI 1.26, 19.13,
𝑃= 0.02) and small cigar (OR = 2.98, 95% CI 1.19, 7.42,i Journal of Environmental and Public Health Journal of Environmental and Public Health 4 Table 1: Demographic information of young adult cigarette smokers attending the festival. Past 30-day small cigar smoker
(𝑁= 31, %)
Nonsmall cigar smokers
(𝑁= 90, %)
Total sample
(𝑁= 121, %)
P value
Country of residence
United States
58.1
55.6
47.6
Outside of United States
41.9
44.4
52.4
Age
18–24 years
51.6
48.9
42.6
25–30 years
48.4
51.1
43.4
≥31 years
0.0
0.0
14.0
Sex (% male)
58.1
54.4
55.4
Race (respondents were able to select
more than one, thus total >100%)
Asian/Pacific Islander
9.7
6.7
7.4
American Indian/Alaskan
Native
3.2
1.1
1.2
Black
22.6
5.6
9.9
∗∗
Hispanic
13.3
12.9
13.2
White
67.7
77.8
75.2
Other
3.2
1.1
3.4
∗P < .05, ∗∗P < .01, ∗∗∗P <.001. Table 1: Demographic information of young adult cigarette smokers attending the festival. Table 2: Multivariate logistic regression analyses predicting small cigar susceptibility. Predictor
OR (95% CI) for intention to smoke
celebrity endorsed small cigar
P
OR (95% CI) for intention to smoke
other small cigars
P
Current cigarette use
—
2.55 (0.94, 6.89)
Current cigar use
5.08 (1.23, 21.08)
∗
4.22 (1.05, 16.93)
∗
Current small cigar use
3.30 (1.24, 8.74)
∗
17.55 (3.87, 79.69)
∗∗∗
Exposure to celebrity-endorsed small cigar
advertisements
3.64 (1.06, 12.54)
∗
1.99 (0.81, 4.95)
∗P < .05, ∗∗P < .01, ∗∗∗P <.001. Table 2: Multivariate logistic regression analyses predicting small cigar susceptibility. 3.5. Susceptibility to Smoke Other Small Cigars in the Future. Over half (53.7%) of our respondents intended to smoke
other small cigars in the future; 41.5% were already current
small cigar smokers. Bivariate analyses found that past 30-
day cigarette (𝑃= 0.06), cigar (OR = 3.75, 95% CI 1.00,
14.00, 𝑃< 0.05), and small cigar smoking (OR = 16.34,
95% CI 3.65, 73.18, 𝑃< 0.001) were significantly associated
and exposure to other small cigar advertisements (𝑃
=
0.06) was marginally associated with the susceptibility to
smoke other small cigars. Table 2 shows the results of our
multivariate model for susceptibility to other small cigars. Past 30-day cigar and small cigar smoking were significantly
associated with susceptibility to smoke small cigars in the
future. Exposure to other small cigar advertisements was not
significantly associated with susceptibility to smoke other
small cigar products, however. 3. Results Of the respondents who were exposed to the
Executive Branch advertisements, 82.4% said seeing the
advertisement made them want to try the product. 2.4.5. Urge to Smoke Small Cigars. Urge to smoke Executive
Branch small cigars was assessed by asking respondents how
much they wanted to smoke the product after exposure to
Snoop Dogg’s advertisement. The urge to smoke variable is
similar to the one used by Sargent and colleagues [25]. A four-
point response category was used to assess urge to smoke
and included 1 = strongly agree to 4 = strongly disagree. We
assessed exposure to and urge to smoke other small cigar
products with similar questionnaire items, by replacing the
phrasing “celebrity-endorsed small cigar product” with “other
small cigar products.’’ Approximately 60.2% of our respondents had been
exposed to other small cigars advertisements. Respondents
reported seeing advertisements for the other small cigars in
magazines (35.1%), on the Internet (23.0%), in convenience
stores or bodegas (12.4%), in smoke shops (6.6%), and on
social media sites (5.4%). Over half of respondents (55.1%)
wanted to try the small cigars after being exposed to the
advertisements. 2.5. Analysis. Respondent demographics, smoking-related
behaviors, advertising exposure, and urge to smoke variables
were explored using descriptive statistics. We assessed the
prevalence of the past 30-day small cigar smoking among
our sample of young adult cigarette smokers. To understand
the demographic and tobacco use characteristics of small
cigar users in our sample, we conducted descriptive analyses
comparing survey responses from those young adult cigarette
smokers who reported past 30-day use of small cigars to those 3.4. Susceptibility to Smoke Executive Branch Small Cigars. Forty-five percent (45.4%) of 121 young adult cigarette
smokers in our sample reported an intention to smoke
Executive Branch small cigars in the future; of these, 40.9%
were current small cigar smokers. Bivariate and multivariate
logistic regression analyses were conducted to assess whether 4 Journal of Environmental and Public Health 4. Discussion Small cigar smoking is a growing public health concern
among young adults, particularly for those with a history
of tobacco use. Among our sample of young adult cigarette Journal of Environmental and Public Health 5 An important limitation of this study is its generalizabil-
ity, as the survey sample largely consisted of a sample of
young adult festival attendees in the United States. Findings
from this study may not generalize to other young adult
populations. While we broadly assessed other small cigars,
this study specifically examined exposure to Snoop Dogg’s
advertisements and promotions of Executive Branch small
cigars. As such, our findings may not generalize to other
small cigar brands. We used brand-specific items to estimate
small cigar use among our respondents. Though these items
may estimate small cigar use more accurately [24], their
use may also limit comparability to studies that used other
measures to capture small cigar use (i.e., single “catch all”
question assessing cigar product use). Finally, the items that
assessed small cigar use in our study did not specifically
ask if participants were smoking tobacco in the small cigar. Despite the use of this item to measure small cigar smoking
in other studies [8], its use may bias findings because it may
also capture use of marijuana. Thus, our findings should be
interpreted with caution. smokers, over 25% concurrently smoked small cigars in the
past 30 days. Our finding is consistent with that of prior
studies [6, 7]. Concurrent use of both products is a public
health concern, as it may increase young adults’ risk for
developing nicotine dependence [2, 26], may lead to an
escalation of tobacco use, and may make it difficult to quit
smoking and achieve long-term abstinence.i Our preliminary findings suggest that exposure to small
cigar advertisements may explain, in part, young adults’
awareness of small cigars. Over 60% of our respondents
reported exposure to other small cigar advertisements, and
14% reported exposure to Snoop Dogg’s Executive Branch
small cigar advertisement. Although traditional outlets such
as magazines and convenience stores were also sources of
advertisement exposure, respondents reported exposure to
small cigar advertisements on the Internet, particularly on
social media sites. This is consistent with growing body of
evidence that has found protobacco images and references
[27–29], in particular small cigar smoking [12, 30] on Internet
social media sites. 4. Discussion We have little evidence to support the claim that the
tobacco industry paid for or supported the hip-hop artist’s
endorsement of this small cigar product. However, the
Federal Trade Commission Report of 1999 documented the
cigar industry’s payments to celebrities for their endorse-
ments, appearances, and cigar product placements. It is
well documented that cigarette smoking has been promoted
through rap and hip-hop music [15, 31] and through music-
themed campaigns (i.e., the Kool Mixx campaign by Brown
& Williamson [13], and concerts and festivals such as Camel’s
“Smooth Moves” and “Speakeasy” tours) [32]. Rap and
hip-hop music genres have a wide reach. Hip-hop artists
appeal to individuals across gender, nationalities (USA versus
international), racial/ethnic backgrounds and ages (although
as noted previously, to youth in particular). These artists
are often viewed as trendsetters. Taken together, the artists’
endorsement of a cigar product may establish brand loyalty
and influence the smoking behaviors of large segments of
their fans. Perhaps the use of celebrity rap and hip-hop artists
to promote small cigar products and smoking is a strategy
that is being used by the cigar industry to circumvent tobacco
advertising restrictions. 4.1. Conclusion. Our study documented young adult smok-
ers’ exposure to a celebrity hip-hop artist’s advertising of
Executive Branch small cigars. We also provided preliminary
evidence of an association between exposure to a celebrity
artist’s small cigar promotion and susceptibility to smoke
the brand-specific small cigar. Our findings indicate that
celebrity endorsement is a potentially important source of
marketing small cigar products to young adults and should
be monitored. The authors declare that they have no conflict of interests. The authors declare that they have no conflict of interests. Journal of Environmental and Public Health 6 [7] A. Richardson, H. Xiao, and D. M. Vallone, “Primary and dual
users of cigars and cigarettes: profiles, tobacco use patterns and
relevance to policy,” Nicotine & Tobacco Research, vol. 14, no. 8,
pp. 927–932, 2012. [24] J. J. Terchek, E. M. G. Larkin, M. L. Male, and S. H. Frank,
“Measuring cigar use in adolescents: inclusion of a brand-
specific item,” Nicotine and Tobacco Research, vol. 11, no. 7, pp. 842–846, 2009. [25] J. D. Sargent, M. Morgenstern, B. Isensee, and R. Hanewinkel,
“Movie smoking and urge to smoke among adult smokers,”
Nicotine and Tobacco Research, vol. 11, no. 9, pp. 1042–1046,
2009. [8] K. Sterling, C. J. Berg, A. N. Thomas, S. A. Glantz, and J. S. Ahl-
uwalia, “Factors associated with small cigar use among college
students,” American Journal of Health Behavior, vol. 37, no. 3, pp. 325–333. [9] Federal Trade Commission, “Cigar sales and advertising and
promotional expenditures for calendar years 1996 and 1997,”
1999, http://www.ftc.gov/os/1999/07/cigarreport1999.htm. [26] F. Baker, S. R. Ainsworth, J. T. Dye, C. Crammer, M. Thun,
and D. Hoffmann, “Health risks associated with cigar smoking,”
Journal of the American Medical Association, vol. 284, no. 18, pp. 735–740, 2000. [10] E. Ramirez,
“Snoop
dogg
launches
new
cigar
brand,”
Billboard, 2012, http://www.billboard.com/articles/columns/
the-juice/512786/snoop-dogg-launches-new-cigar-brand#/col-
umn/the-juice/snoop-dogg. [27] M. V. Carroll, A. Shensa, and B. A. Primack, “A comparison
of cigarette- and hookah-related videos on YouTube,” Tobacco
Control, vol. 22, no. 5, pp. 319–323, 2013. [11] Snoop dogg launching cigar brand at festival, “Rapper will
introduce executive branch smokes at coachella,” Rolling Stone,
2012,
http://www.rollingstone.com/music/news/snoop-dogg-
launching-cigar-brand-20120112. [28] S. R. Forsyth and R. E. Malone, ““YouTube” telling or selling
you something? Tobacco content on the YouTube video-sharing
website,” Nicotine and Tobacco Research, vol. 12, no. 8, pp. 810–
816, 2010. [12] A. Richardson, O. Ganz, and D. Vallone, “The cigar ambassador:
how Snoop Dogg uses Instagram to promote tobacco use,”
Tobacco Control, 2013. [29] B. Freeman and S. Chapman, “Is “YouTube” telling or selling
you something? Tobacco content on the YouTube video-sharing
website,” Tobacco Control, vol. 16, no. 3, pp. 207–210, 2007. [13] N. Hafez and P. M. Ling, “Finding the kool mixx: how
Brown and Williamson used music marketing to sell cigarettes,”
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vehicle for little cigars and cigarillos?” Tobacco Control, 2012. [31] R. H. DuRant, E. S. References [1] L. T. Kozlowski, K. M. Dollar, and G. A. Giovino, “Cigar/cigaril-
lo surveillance. Limitations of the U.S. department of agricul-
ture system,” American Journal of Preventive Medicine, vol. 34,
no. 5, pp. 424–426, 2008. [2] D. Hoffmann and I. Hoffman, “Chemistry and toxicology. Cigars: health effects and trends smoking and tobacco control
monograph No 9. U.S. DHHS,” National Institutes of Health,
National Cancer Institute, pp. 55–104, 1998. g
Exposure to the hip-hop artist’s small cigar advertise-
ments appeared to influence small cigar smoking susceptibil-
ity among the young adult cigarette smokers in our sample. Those who reported exposure to the artist’s Executive Branch
small cigar advertisements were three times more likely to
intend to smoke these small cigars in the future. Notably,
over 40% of those who reported an intention to use the
Executive Branch small cigars were already past 30-day small
cigar smokers. Although not previously documented for
music entertainment, our findings are consistent with studies
that found an association between exposure to prosmoking
depictions in films and smoking behavior among adolescents
[15]. Future studies that examine the association between
exposure to small cigar advertisements and intention to
smoke among other young adult smoking samples are needed
to substantiate our findings. [3] M. D. Blank, A. Nasim, A. Hart Jr., and T. Eissenberg, “Acute
effects of cigarillo smoking,” Nicotine and Tobacco Research, vol. 13, no. 9, pp. 874–879, 2011. [4] L. A. Fabian, L. L. Canlas, J. Potts, and W. B. Pickworth, “Ad lib
smoking of black & mild cigarillos and cigarettes,” Nicotine and
Tobacco Research, vol. 14, no. 3, pp. 368–371, 2012.l [5] A. K. Regan, S. R. Dube, and R. Arrazola, “Smokeless and fla-
vored tobacco products in the U.S.: 2009 styles survey results,”
American Journal of Preventive Medicine, vol. 42, no. 1, pp. 29–
36, 2012. [6] J. M. Rath, A. C. Villanti, D. B. Abrams, and D. M. Vallone,
“Patterns of tobacco use and dual use in US young adults: the
missing link between youth prevention and adult cessation,”
Journal of Environmental and Public Health, vol. 2012, pp. 1–9,
2012. Journal of Environmental and Public Health Rome, M. Rich, E. Allred, S. J. Emans, and
E. R. Woods, “Tobacco and alcohol use behaviors portrayed in
music videos: a content analysis,” American Journal of Public
Health, no. 7, pp. 1131–1135, 1997. [14] U. S. Department of Health and Human Services, “Preventing
tobacco use among youth and young adults (2012): a report of
the surgeon general,” Tech. Rep., Rockville, MD: Office of the
Surgeon General, 2012. [32] C. R. Stanton, A. Chu, J. Collin, and S. A. Glantz, “Promoting
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British American tobacco and the Ministry of Sound,” European
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in US cigar use epidemiology among young adults aged 18–25
years: a focus on race/ethnicity and brand,” American Journal of
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rette advertisements: links between exposure, liking, and the
appeal of smoking,” Tobacco Control, vol. 7, no. 2, pp. 129–133,
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“Seeing, wanting, owning: the relationship between receptivity
to tobacco marketing and smoking susceptibility in young
people,” Tobacco Control, vol. 7, no. 2, pp. 123–128, 1998. [19] D. M. Straub, N. K. Hills, P. J. Thompson, and A.-B. Moscicki,
“Effects of pro- and anti-tobacco advertising on nonsmoking
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pro-tobacco and anti-tobacco messages on adolescent smoking
susceptibility,” Nicotine and Tobacco Research, vol. 8, no. 3, pp. 455–465, 2006. [21] I. Ajzen and M. Fishbein, Understanding Attitudes and Predict-
ing Social Behavior, Prentice Hall, Englewood Cliffs, NJ, USA,
1980. [22] Coachella Breaks Attendance Record, “NBC Southern Califor-
nia,” 2013, http://www.nbclosangeles.com/news/local/Coach-
ella-Breaks-Attendance-Record-85000-stabbing-148020395
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NADPH Oxidase Biology and the Regulation of Tyrosine Kinase Receptor Signaling and Cancer Drug Cytotoxicity
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Int. J. Mol. Sci. 2013, 14, 3683-3704; doi:10.3390/ijms14023683
OPEN ACCESS
International Journal of
Molecular Sciences
ISSN 1422-0067
www.mdpi.com/journal/ijms
Review
NADPH Oxidase Biology and the Regulation of Tyrosine Kinase
Receptor Signaling and Cancer Drug Cytotoxicity
Rafael Paletta-Silva 1,†,*, Nathália Rocco-Machado 2,3,†,* and JoséRoberto Meyer-Fernandes 2,3,*
1
2
3
†
Clinical Research Coordination, Nacional Institute of Cancer (INCA), AndréCavalcanti Street, 37,
Rio de Janeiro, RJ 20231-050, Brazil
Institute of Medical Biochemistry, Federal University of Rio de Janeiro (UFRJ), CCS, Bloco H,
University City, Fundão Island, Rio de Janeiro, RJ 21941-590, Brazil
Institute of National Science and Technology of Structural Biology and Bioimage (INCTBEB),
CCS, Bloco H, University City, Fundão Island, Rio de Janeiro, RJ 21941-590, Brazil
These authors contributed equally to this work.
* Authors to whom correspondence should be addressed; E-Mails: rafael.paletta@hotmail.com (R.P.-S.);
nathaliarocco@hotmail.com (N.R.-M.); meyer@bioqmed.ufrj.br (J.R.M.-F.);
Tel.: +55-21-2562-6781; Fax: +55-21-2270-8647.
Received: 24 December 2012; in revised form: 28 January 2013 / Accepted: 31 January 2013 /
Published: 7 February 2013
Abstract: The outdated idea that reactive oxygen species (ROS) are only dangerous
products of cellular metabolism, causing toxic and mutagenic effects on cellular
components, is being replaced by the view that ROS have several important functions in
cell signaling. In aerobic organisms, ROS can be generated from different sources,
including the mitochondrial electron transport chain, xanthine oxidase, myeloperoxidase,
and lipoxygenase, but the only enzyme family that produces ROS as its main product is the
NADPH oxidase family (NOX enzymes). These transfer electrons from NADPH
(converting it to NADP−) to oxygen to make O2•−. Due to their stability, the products of
NADPH oxidase, hydrogen peroxide, and superoxide are considered the most favorable
ROS to act as signaling molecules. Transcription factors that regulate gene expression
involved in carcinogenesis are modulated by NADPH oxidase, and it has emerged as a
promising target for cancer therapies. The present review discusses the mechanisms by
which NADPH oxidase regulates signal transduction pathways in view of tyrosine kinase
receptors, which are pivotal to regulating the hallmarks of cancer, and how ROS mediate
the cytotoxicity of several cancer drugs employed in clinical practice.
Int. J. Mol. Sci. 2013, 14
3684
Keywords: NADPH oxidase; reactive oxygen species; cancer; tyrosine kinase receptors;
cancer drugs
1. Introduction
Reactive oxygen species (ROS) are formed during aerobic metabolism and are much more reactive
than molecular oxygen. For many years, they were only associated with cell injury, but currently, they
are viewed as important components of the biological processes [1–5]. In aerobic organisms, ROS can
be generated from different sources, including the mitochondrial electron transport chain, xanthine
oxidase, myeloperoxidase, lipoxygenase, and uncoupled endothelial NO synthase, but the only enzyme
family that produces ROS as its main product is the nicotinamide adenine dinucleotide phosphate
(NADPH) oxidase family (NOX enzymes) [6]. The NOX family comprises seven members: NOX 1–5
and the dual oxidases (DUOXs) DUOX 1 and DUOX 2. The NADPH oxidase complexes were
initially thought to be specific to phagocyte cells, but these enzymes are now recognized as being
almost universally expressed in nonphagocytic cells.
Currently, NADPH oxidase-derived ROS are known to modulate a variety of functions in different
cell types. There is increasing interest in the investigation of redox-sensitive signaling pathways, as
cancer cells present high oxidative metabolism with high ROS content. Such signals involve
transcription factors, such as AP-1 and NF-kappaB, and cell signaling components, such as p38
MAPKs, PKC and PI3K/AKT, which are all NADPH-derived ROS sensitive. Our goal in this review
is to discuss the mechanisms by which NADPH oxidase regulates signal transduction pathways in
view of tyrosine kinase receptors, a pivotal regulator of hallmarks of cancer, and the role of NADPH
oxidase in the mechanisms of action of cancer drugs.
2. The NOX Family
The NOX family consists of seven members: NOX 1–5 and two dual oxidases (DUOXs), DUOX 1
and DUOX 2. All NOX isoforms have six transmembrane domains in the N-terminal half, four
conserved histidine residues located in the third and fifth transmembrane helices, which coordinate two
hemes, and a flavoprotein domain and NADPH-binding domain in the C-terminal cytosolic region.
NOX 5 also has Ca2+-binding domains, EF-hands, at the N-terminus. In addition to EF-hands, DUOX
enzymes have a membrane-spanning region and a peroxidase-like domain at the N-terminus [6,7].
Figure 1 and Table 1 show the seven isoforms of NADPH oxidases, their subunits and regulators.
Different NOX proteins are often expressed in the same cell or tissue. Although they produce the
same type of ROS, the enzymes regulate distinct functions in different cell types. These findings
suggest that the complement of NOX proteins within a cell and their subcellular localization and
coupling to external stimuli are determinants of the response to NOX activation.
Int. J. Mol. Sci. 2013, 14
3685
Figure 1. Schematic sequence of NOX isoforms and their regulatory subunits.
Table 1. NOX isoforms, their regulatory subunits and activators.
NOX isoforms
NOX1
NOX2
NOX3
NOX4
NOX5
DUOX1
DUOX2
Subunits
p22phox, NOXA1, NOXO1 and RAC1
gp91phox, p22phox, p40phox, p47phox,
p67phox, RAC1
p22phox, NOXO1, NOXA1, RAC1
P22phox
NONE
DUOXA1, DUOXA2
DUOXA1, DUOXA2
Regulators
ANG II, PDGF
PKC, (TNF)-α,
phosphatidic acid
Unknown
Poldip2
2+
Ca , ptdlns(4,5)p2
IL-4, IL-3,Camp, PKA
IFN-γ, PLC, PKC
References
[8,9]
[7,8,10–16]
[8,17–19]
[20,21]
[7,8,22]
[7,23–25]
[7,23–25]
2.1. NOX 2
The classical NADPH oxidase in phagocytes was the first member of the family to be described [26].
It consists of two membrane-bound elements, gp91phox (“phox” stands for phagocyte oxidase) and
p22phox; three cytosolic proteins, p40phox, p47phox, p67phox; and a small G-protein, Rac [10].
Gp91phox is the catalytic subunit, also called NOX 2, and has a C-terminal cytosolic region that
contains a flavoprotein domain, which is homologous to flavoprotein dehydrogenase flavin adenine
dinucleotide (FAD)-binding sequences and a sequence that represents a NADPH-binding site [8,27].
Protein kinase C (PKC) has been shown to be involved in the phosphorylation of gp91phox during the
activation of human neutrophils [11]. This subunit forms a heterodimer with p22phox at the plasma
membrane, which is called flavocytochrome b558 [6–8].
P22phox has a proline-rich sequence (PRR) that is involved in binding to Src homology 3 (SH3)
domains in the cytosolic subunit p47phox. In the resting state, the SH3 domains of p47phox bind the
autoinhibitory region (AIR) in the C-terminal half, interrupting binding to p22phox. In the presence of
Int. J. Mol. Sci. 2013, 14
3686
an appropriate stimulus, such as phorbol 12-myristate 13-acetate (PMA), serine residues of
p47phox can be phosphorylated through PKC [12], and p47phox becomes available to bind p22phox.
It was also shown that tumor necrosis factor (TNF)-α induces p47phox phosphorylation in human
neutrophils [13]. The p22phox subunit can be phosphorylated by PKC and phosphatidic acid on
threonine residues [7,8,14].
Another cytosolic component, p67phox, is associated with the C-terminal PRR of p47phox through
its C-terminal SH3 domain. It also has four tetratricopeptide-rich regions that are responsible for
binding Rac proteins and a proline-rich region. The C-terminal domain contains the phox and Bem1p
(PB1) domain, whose function is to create heterodimers between proteins containing the PB1 domain.
P67phox is phosphorylated on its serine and threonine residues by PKC-dependent and -independent
pathways [15].
P40phox, which is also localized in the cytosol, is associated with p67phox via mutual phox and
PB1 domains. It can be phosphorylated at the serine 315 and threonine 154 residues by PKC [16], and
it is not necessary to activate the oxidase complex.
The translocation of cytosolic components and their association with the membrane subunit lead to
the activation of the enzyme [7,8]. NOX 2 is essential in innate host defense by both producing ROS to
attack invaders and acting as a signaling molecule to initiate inflammatory and immunoprotective
responses [28]. It is expressed not only in phagocytes but also vascular cells [29], the endothelium,
fibroblasts, cardiomyocytes, skeletal muscle, hepatocytes, and hematopoietic stem cells [30]. NOX 2
activity dysregulation may lead to dysfunction and contribute to hypertension [17,31].
2.2. NOX 1
NOX 1 was the first homologue of gp91phox to be described, and it was originally isolated from
colon epithelial cells (CECs) [32,33] but is also found at lower levels in vascular smooth cells,
endothelial cells, the uterus, placenta, prostate, osteoclasts, retinal pericytes and macrophages [8,9,29].
There are two proposed roles for the physiological action of NOX1 in CECs: immune defense and cell
proliferation. From a pathological view, the misregulation of the enzyme could be responsible for
inflammatory bowel disease and carcinogenesis [17,34,35].
This isoform is composed of the membrane subunit p22phox; two cytosolic subunits, the 67phox
homologue NOXA1 (NOX activator 1) and the p47phox homologue NOXO1 (NOX organizer 1); and
Rac1. Its expression is induced by vasoactive factors, such as angiotensin II (Ang II) and
platelet-derived growth factor (PDGF) [8,9].
2.3. NOX 3
NOX 3 was discovered in 2000 [36], and it is expressed in the inner ear, lung endothelial cells and
fetal spleen, kidney, lung, and skull, suggesting that NOX3 plays an important role in tissue
development but is turned off in adult tissue. The activity of this subunit depends on p22phox,
NOXO1, NOXA1 and Rac1 [8,17–19].
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2.4. NOX 4
NOX 4 was first identified in the kidney [37,38] but was subsequently found in many cell types,
such as smooth cells, endothelial cells, fibroblasts, keratinocytes, osteoclasts, neurons, and
hepatocytes. NOX 4 is constitutively and highly expressed in various cell lineages [17,29].
This isoform requires only the p22phox subunit to exert its enzymatic activity [20]; however, unlike
NOX 1, NOX 2 and NOX 3, it does not require the organizer subunit-interacting proline-rich domain
of p22phox [39]. Recently a p22phox-interacting protein, polymerase delta-interacting protein
(Poldip2), was described to enhance the activity of NOX 4 [21]. The enzyme has been suggested to be
involved in stress signals in the kidney and smooth muscle, TGFβ-induced differentiation, insulin
signalling, oxygen sensing, cardiac differentiation and transcriptional regulation [17].
2.5. NOX 5
NOX 5 is a calcium (Ca2+)-dependent homologue, and it has four Ca2+-binding domains (EF-hands)
in the N-terminus. This isoform has been identified in lymphoid tissues, testes, the spleen, and
endothelial cells. Unlike other NOX proteins, NOX 5 does not need any other subunit for its
activation; however, the enzyme is regulated by intracellular Ca2+ levels [7,8]. It was reported that
phosphatidylinositol (4,5)-bisphosphate (Ptdlns(4,5)p2) plays a key role in the translocation of NOX 5
to the plasma membrane [22]. NOX 5 can also be activated by PKC through the phosphorylation of
serine residues. This activation process has no equivalent in the NOX family and seems to occur on
regions of this enzyme that are different from other isoforms of NADPH oxidase [40].
This enzyme participates in endothelial cell proliferation, migration and angiogenesis [8]. NOX 5
has been observed in certain pathologies, such as hairy cell leukemia [41], melanoma cells [42],
prostate cancer cells [43], and Barrett’s mucosa [44].
2.6. DUOX 1 and DUOX 2
The dual oxidases DUOX 1 and DUOX 2 were first described as homologues of NOX 2 in the
thyroid gland [45,46]. They are composed of a NOX-like region in the C-terminal half, two EF-hands,
a membrane-spanning region and a prolonged peroxidase-like N-terminus. The name dual oxidase is
based on the presence of an extracellular peroxidase-like domain in addition to their NOX-like
NADPH oxidase core; however, no clear peroxidase activity has been attributed to human
DUOXs [23,47].
DUOXs do not require other components of NADPH oxidase for their activity, but they are Ca2+
dependent and also need the DUOX maturation factors DUOXA1 and DUOXA2. DUOXAs are
transmembrane glycoproteins characterized as endoplasmic reticulum (ER) proteins that are essential
for ER-to-Golgi transition, maturation, and targeting DUOX 1 and DUOX 2 to the plasma membrane
as functional complexes [7,24]. DUOX 1 is expressed in the thyroid gland, airway epithelia, placenta,
prostate, testis, pancreas and heart. DUOX 2 is expressed in the thyroid gland, airway epithelia,
epithelial cells in salivary excretory ducts and rectal glands [7].
In the airway epithelia, DUOX 1 gene expression is induced by Th2 cytokines (IL-4 and IL-13),
and DUOX 2 gene expression is induced by Th1 cytokines (IFN-γ) [48]. Differential DUOX regulation
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might be important for host defense and inflammatory responses [7]. Mutations in the DUOX 2 gene in
the thyroid have reported in congenital hypothyroidism [49]. At the post-translational level, both
enzymes are regulated by different intracellular signaling cascades. DUOX 1 responds to cAMP and
PKA-mediated phosphorylation, and DUOX2 is stimulated by the PLC cascade and PKC-dependent
phosphorylation [25].
3. Reactive Oxygen Species
ROS are produced naturally or through metabolic dysfunction. They are responsible for
phagocytosis and the regulation of cellular growth, signaling and the synthesis of important substances.
However, in excess, they can be detrimental.
Superoxide (O2•−), the main ROS produced in vivo, is highly reactive and short lived. Therefore, it
has to be produced in proximity to its target to be effective as a signaling molecule. O2•− is capable of
reacting with nitric oxide (NO), forming peroxynitrite (OONO−), a strong oxidizing, nitrating and
nitrosylating agent. O2•− also reacts with (FeS)4 clusters within proteins, which may release ferric irons,
and protein thiols, such as cysteine residues. O2•− is not able to cross biological membranes due to its
negative charge and can be dismutated to hydrogen peroxide (H2O2) spontaneously or enzymatically
via superoxide dismutase (SOD) [17,50,51]. H2O2 is able to cross biological membranes, is more
stable than O2•− and is regulated by catalase and glutathione (GSH) peroxidase, which converts H2O2
to water and other metabolites. The reaction of H2O2 with chloride (Cl−), catalyzed by
myeloperoxidase, produces the oxidant hypochlorite (ClO−). Reaction with metal ions, such as copper
(Cu+) and iron (Fe2+), generates hydroxyl radical (·
HO), which reacts rapidly and indiscriminately with
biomolecules of all classes. H2O2 can also reversibly react with cysteine residues on proteins, leading
to their activation or inactivation, directly activate redox-sensitive kinases, such as protein kinase B,
protein kinase C, the mitogen-activated protein kinase family and Janus kinase, and have a direct effect
on ion channels or receptors [17,51,52].
3.1. NOX-Derived ROS
As mentioned earlier, NADPH oxidases produce ROS as their main product and not as a
consequence, as in the case of other sources, such as the mitochondrial electron transport chain,
xanthine oxidase, myeloperoxidase, lipoxygenase and uncoupled endothelial NO synthase, which
produce ROS as a side effect of their action. The existence of many NOX isoforms suggests the
relevance of redox-sensitive signaling cascades [51,53,54]. NADPH oxidases produce ROS by
transferring an electron from NADPH to oxygen via FAD and the two heme groups, with the second
heme group being responsible for reducing molecular oxygen. Heme is an obligate electron donor;
therefore, it is generally accepted that O2•− is the initial product of NADPH oxidases, although it was
previously reported that NOX4 and DUOXs directly produce hydrogen peroxide (H2O2) [24,55,56].
Studies on NOX4 ROS production are controversial. In Serrander et al. [40], NOX4 induced a
strong signal with probes that detected extracellular H2O2 but not with probes that detected
extracellular O2•−. These authors also used the DHE method, which is widely used to detect
intracellular O2•−, and no NOX4 signal was detected. Another method to measure intracellular O2•−, the
reduction of NBT to formazan, was also used. Induced NOX4 cells elicited significant NBT reduction
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compared with non-induced cells. As only O2•− is capable of reducing NBT, this result provides strong
support for NOX4 O2•− production. The study suggested that NOX4 produces primarily O2•− in a tight
intracellular compartment devoid of DNA, and this O2•− is rapidly converted into H2O2. Others believe
that NOX4 produces primarily H2O2. Dikalov et al. [57] showed that siRNA against NOX4 does not
reduce O2•− but does reduce H2O2 production, and Takac (2011) [56] identified the external E-loop of
the protein as an essential structure for H2O2 production and showed that alterations of the E-loop
switch in NOX4 led to O2•− production.
DOUX1 and DUOX2 were originally found to produce mainly H2O2 [58]. No homology was found
in the DUOXs with the E-loop of NOX4, but they have a long NH2 extracellular domain that might be
involved in H2O2 production [24]. The DUOX maturation factors have also been associated with the
type of ROS produced. When DUOX1 and DUOX2 are coexpressed with DUOX, they are not retained
as ER resident proteins. The DUOX-DUOXA complex migrates to the plasma membrane and
produces high amounts of H2O2 but no detectable superoxide [55]. Alterations of DUOX maturation
factors switch the enzyme from H2O2 to O2•− formation [24,56].
4. Regulation of Cancer Cell Biology by NADPH Oxidase Activity: Implications in Hallmarks
of Cancer
The role of ROS-dependent NADPH oxidase in biological systems can be classified into at least
two functions: (1) promoting oxidative stress; an imbalance between the generation and neutralization
of ROS in cells has deleterious effects on macromolecules, such as proteins, nucleic acids and lipids,
leading to cellular damage and enhancing the risk of mutations; and (2) the regulation of several
signaling pathways that are redox sensitive contributes to cancer pathophysiology [59,60].
ROS-derived NADPH oxidase activity is closely related to several mechanisms underlying cancer
cell biology and, consequently, disease progression. The high metabolic oxidative stress observed in
cancer cells modulates a wide range of processes that confer acquired capabilities during tumor
development. It has been demonstrated that all the hallmarks of cancer that were classified by Hanahan
and Weinberg [61] and reviewed [62] (e.g., sustaining proliferative signaling, evading growth
suppressors, resisting cell death, enabling replicative immortality, inducing angiogenesis, activating
invasion and metastasis, reprogramming energy metabolism, and evading immune responses) can be
modulated by ROS derived from NADPH oxidase isoform activities in a redox-regulated manner.
NOX dictates carcinogenesis through the regulation of several cell signaling pathways related to
carcinogenesis that respond to stress signals, such as Janus kinase-signal transducer and activator of
transcription (JAK-STAT), protein kinase C, mitogen-activated protein kinase (MAPK), AKT [60,63]
(Figure 2).
The effects of NOX on the hallmarks of cancer mentioned above have been well documented in
recent reports [59,64]. In the following sections, we will present and discuss the role of NOX enzymes
in cancer disease in view of tyrosine kinase receptor (TKR) signaling, which has been implicated in
pivotal hallmarks of cancer and is currently being explored as cancer molecular target therapy.
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Figure 2. Reactive oxygen species (ROS) can act in a synergic manner with tyrosine
kinase receptor (TKR) to promote carcinogenesis. TKR can enhance the
expression/activity of NOX isoforms. The ROS generated from NOX activity can sustain
the activation of TKR signaling pathways and of transcription factors involved on
carcinogenesis through inhibition of PTP activities.
5. Redox-Sensitive Regulation of Signaling Pathways: NADPH-Derived ROS as Mediators
Thiol redox regulation dictates gene expression, interfering with transcription factor activities, and
can also act as a second messenger, interfering with protein functions responsible for transducing cell
signaling events. Thiol group modifications are thought to play a central role in the cellular signaling
process through ROS modifications. In general, the mechanisms underlying such regulation are dictated
by redox reactions, which alter the cysteine residue state in protein phosphatases and kinases [65].
TKRs are one of the most important redox-responsive signaling cascades that are commonly
enhanced in several cancers cells compared with normal cells. Enhanced signal transduction caused by
growth factors, such as epidermal growth factor (EGF) and platelet-derived growth factor (PDGF),
cytokines and hormones results in an uncontrolled proliferative response. These events are involved in
the enhancement of the signaling response through an overall increase in tyrosine phosphorylation and
inhibition of protein tyrosine phosphatases (PTPs) by oxidation [66]. In this context, it has been
accepted that ROS derived from NADPH oxidase activity is closely related to enhancing signal
transduction engaged by the activation of TKRs or inhibiting tyrosine phosphatase activity, which is
responsible for inactivating TKRs or TKR-associated proteins.
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5.1. NOX Modulates PTP Activities: Maintaining TKR Signaling by ROS
PTPs contain an essential cysteine residue that is responsible for interacting with phosphate as
mechanisms of catalysis (dephosphorylating). This residue is rapidly oxidized by hydrogen peroxide,
resulting in reversible enzyme inactivation. As cancer cells have high oxidative metabolism, in most
cases, PTPs are in an inactive state, contributing to the maintenance of TKR activation and
consequently promoting carcinogenesis events.
NOX regulates the activity of PTP1B, PTEN, SHP1/2 and low-molecular-weight protein tyrosine
phosphatase (LMW-PTP) [59] and sustains TKR phosphorylation and the activation of EGFR
downstream proteins. PTEN is involved in regulating a variety of cellular functions, including the cell
cycle, apoptosis, DNA repair, signal transduction and cell adhesion [67]. PTEN inactivation by
hydrogen peroxide occurs by oxidizing Cys-124 in the catalytic site followed by the formation of a
disulphide with Cys-71 [68]. Hydrogen peroxide-derived NOX1 inactivates PTEN activity. As PTEN
dephosphorylates PIP3 to PIP2, PIP3 accumulation sustains cell signal transduction mediated by
PIP3/AKT [69]. In prostate cancer, hydrogen peroxide enhances migration and invasion by
inactivating PTEN and increasing the expression of the chemoattractant receptor CXCR-4 [70]. PTPs
are also involved in anti-apoptotic effects and prosurvival events; in pancreatic cancer cells,
low-molecular-weight phosphatases are inhibited by NOX4, leading to sustained activation of the
JAK2 pathway [71]. Leukemic B cells (hairy cell cancer) express NOX5 and produce ROS in a
calcium-dependent manner that does not involve PKC. Furthermore, NOX5 colocalization with SHP-1
inhibits phosphatase activity, which contributes to maintaining cells in active states by sustaining
phosphorylation of the CD22 receptor [41].
6. Interplay between Tyrosine Kinase Receptor Signalling and ROS-Derived NADPH in
Regulating Carcinogenesis
6.1. Epidermal Growth Factor Receptor (EGFR)
EGFR is a member of the ErbB family of tyrosine kinase receptors and is commonly up-regulated
in non-small cell lung cancer (NSCLC), head and neck cancer, colorectal cancer and hepatocellular
cancer. In general, EGFR mutation is correlated with a poor clinical prognosis. The molecular analysis
of EGFR gene mutations is used as a guideline protocol to determine the prescription of EGF signaling
tyrosine kinase inhibitors, such as erlotinib, and monoclonal antibodies (mAbs) against EGFR, such as
cetuximab (chimeric mAb) and panitumumab (humanised mAb), as single agents or associated with
standard chemotherapy and/or radiotherapy protocols [72,73].
The enhanced EGFR signaling observed in EGFR-mutated cancer cells mediates the hallmarks of
cancer in different cell types through the activation of pro-survival signaling pathways, including
PI3K/Akt, MAPK and JNK [74]. Several studies have reported that NOX is upstream of those signals.
NOX1 enhances the expression of EGFR signaling components and confers autocrine growth. Early
hydrogen peroxide-derived NOX1 activation (15 min) is required for the phosphorylation of several
EGFR signaling pathways, such as c-Src/ERKs and p38/AKT (30 min). Interestingly, the activation of
EGFR components is required as a mechanism of NOX-1 regulating its own expression and the
expression of an EGFR ligand (TGF-α) [75]. In leiomyoma smooth muscle cells, the inhibition of
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NADPH oxidase impairs MAPK signaling activation and decreases the cell proliferation response to
EGF [76].
Transforming growth factor beta (TGF-β) is considered to be a liver tumor suppressor because of its
ability to induce apoptosis. A link between TGF/EGFR/NOX signaling has emerged as a mechanism
that dictates cell death or apoptotic cell resistance in liver cancer cells. The activation of MAPK/ERK
(EGFR-related signaling) confers cell resistance to TGF-β-induced apoptosis through impairing
NOX4, which is induced upstream of mitochondrial-dependent apoptotic proteins of the BCL-2
family. NOX4 knockdown impairs ROS increase and all the mitochondrial-dependent apoptotic
features by regulating BIM, BMF, BCL-XL, and MCL1 levels. Inhibiting EGFR potentiates
TGF-β-induced NOX4 up-regulation and increases hepatoma cell death [77–79].
The proto-oncogene RAS belongs to the G-small proteins, and the structurally related genes
H-RAS, N-RAS and K-RAS codify cell signal transducers that are associated with TKRs. K-RAS is
commonly overexpressed (mutated) in several tumors, especially in pancreatic cancer (95% of
cases) [80]. Because patients with K-RAS mutation do not respond to EGFR inhibitors, understanding
the role of RAS in cancer pathophysiology is important, and RAS are being studied as a molecular
target therapy for patients who have RAS gene mutation.
A relationship between RAS and the NOX family has been demonstrated to play an important role
in cancer progression. RAS can stimulate migration and proliferation in a ROS-dependent manner in
colon cancer cells, and RAS signaling upregulates NOX1 expression via the MEK-ERK-GATA-6
pathway. Mutations in the GATA-6-ERK phosphorylation site abolish NOX1 upregulation and impair
cell growth [81].
Rac (an activator of NOX2 and NOX4) activation represents an important downstream effecter of
RAS [82]. As Rac is a potent activator of NADPH oxidase activity, some authors have suggested an
interplay between Rac/NADPH oxidase and K-RAS. Inhibiting Rac1 activity (Rac1 mutant construct)
inhibits the growth of mutant K-RAS human pancreatic cancer cells but not pancreatic cancer cells
with wild-type K-RAS [83] in a superoxide-dependent manner. The induction of NOX activity by
K-RAS was elucidated in pancreatic cancer cell lines; those that overexpressed K-RAS had significantly
higher levels of superoxide than those that did not express the oncogene without diminishing
antioxidant defense (superoxide dismutase). Additionally, it was demonstrated that NOX2 was only
expressed in cells expressing K-RAS and that silencing NOX2 expression diminished superoxide
generation and cell growth [84].
RAS/NOX also modulates cell growth and proliferation via activating the ERK/MAPK signaling
pathway and cyclin D1, a regulator of the cell cycle [85].
The role of RAS and Rho (a crucial small GTPase protein that functions in the formation of stress
fibers and focal adhesions) signaling on invasion/metastasis-related events was also demonstrated in
view of ROS. NOX1 causes the loss of stress fiber formation and suppression of focal adhesion in a
RAS-dependent manner through inhibiting Rho activity. Hydrogen peroxide-derived NOX1 inactivates
LMW PTP (phosphatase), and subsequent p190RhoGAP activation causes Rho downregulation [86].
Additionally, NOX1 inhibition attenuates RAS-dependent metalloproteinase-9 expression via
inhibiting IKKα kinase/NF-κB signaling [87].
The role of NOX in the activation of tyrosine receptors is even more complex due to its ability to
participate in the cross talk between cell surface receptors, such as G-protein-coupled receptors
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(GPCRs) and RTK. GPCRs employ multiple strategies for EGFR transactivation that can be ligand
dependent, which involves the release of the EGFR ligand family (TGF-α and EGF) by the action
of metalloproteases (MMPs), such as TACE/ADAM, and the activation of EGFR in an
autocrine/paracrine manner [88], or ligand independent, through sustaining EGFR activation due to the
activation of protein kinase signaling pathways associated with the receptor. As the aim of the present
review is to focus on effects of the NOX family on signaling events that participate in carcinogenesis,
the complex signaling pathway of GPCRs will not be discussed. For further information, reviews
addressing GPCRs and their signaling in detail have been published [88,89].
The transactivation of EGFR by NOX has been related to premalignant changes in lung cells.
ROS-derived NADPH oxidase activation by tobacco smoke induces metalloproteinase TACE
activation in a Src/PKCε-dependent manner, which induces the processing and release of amphiregulin
and EGFR activation [90]. In colon rectal cancer, c-Src induces the expression of NOX1 in a
Rac1-dependent manner [91]. In NSCLC, EGFR transactivation is also mediated by ROS-dependent
NADPH oxidase activity/c-Src, which results in the downstream activation of the PI3K/AKT/NF-κB
pathway and cyclooxygenase-2, which are molecular predictors of the overall survival of patients with
NSCLC [92].
The human formyl peptide receptor (FPR) and FPR-like 1 (FPRL1) belong to the GPCR family and
mediate the transactivation of EGFR through NOX-dependent ROS generation. In a human lung
adenocarcinoma cell line (CaLu-6), FPRL1 activated NADPH oxidase in an ERK-dependent manner.
Superoxide derived from the NADPH oxidase activation of c-Src (TYR-416), a known EGFR
activator [93], transactivates EGFR/JAK/STAT3, promoting cell proliferation [94].
The bombesin receptor family (BBR) is another GPCR involved in regulating carcinogenesis.
BB2R and BB3R stimulate ERK and EGFR phosphorylation by stimulating metalloproteases and the
release of TGF-α in a c-Src-dependent manner in head and neck cancer cell (HNSCC) lines and
NSCLC in an NADPH oxidase-dependent manner [95].
6.2. Vascular Endothelial Growth Factor Receptor (VEGFR)
The major role of VEGFR is to induce angiogenesis and support glucose uptake in cancer cells. The
induction of new vessel formation by VEGF plays a critical role in supplying the high metabolic
demand of cancer cells. Similar to EGFR, disrupting VEGFR signaling pathways has been commonly
used as molecular target therapy for highly vascularised tumors, such as colorectal and hepatic
cancer [96,97], lung cancer [98], and breast cancer [99].
The induction of VEGF gene expression by hypoxia in tumor cells involves both an increase in the
rate of gene transcription, which is mediated by the transcription factor hypoxia-inducible factor (HIF),
and an enhancement of the expression and stability of VEGF and/or VEGFR mRNA [100,101]. In this
context, NADPH oxidase isoforms present an important feature in regulating new blood vessel
formation downstream of VEGFR signaling and mediating HIF-1 expression in a ROS-dependent
manner. Human microvascular endothelial cells overexpressing Nox4 present an increase in VEGF and
HIF-1 in an insulin-dependent AKT phosphorylation manner. Downregulating NOX4 impairs
endothelial cell migration and proliferation and, consequently, angiogenesis [102].
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In cancer, NOX4-dependent ROS production and HIF-1 activation are observed in the hypoxic
region of glioblastoma tumors [103]. The VEGFR/NOX4-dependent accumulation of HIF-1
surrounding melanoma tumors is correlated with chemotherapy and radiotherapy resistance [104]. In
prostate cancer cell models, PKCδ activation increases HIF and VEGF levels in an NADPH oxidase
dependent-manner, implying a role for ROS-derived NADPH oxidase [105]. The stability of HIF-1
expression is mediated by p22phox-dependent mTOR activation in renal cancer cell models [106].
In K-RAS transformed rat kidney cells, NOX1 mediates angiogenesis, enhancing VEGF expression
through the phosphorylation and activation of a transcription factor, SP-1, which binds to the VEGF
promoter [107]. In leukemic cells, Rac-1 and AKT activate NOX2 and NOX4 in response to VEGF,
and ROS-derived NOX activities sustain VEGFR-2 receptor phosphorylation and support glucose
uptake [108]. In endothelioma cells, VEGF stimuli upregulate NOX1, but not NOX2 or NOX4. NOX1
expression mediates angiogenic effects through activating NF-κB and inhibiting PPARα [109], which
inhibits pro-angiogenic effects by blocking VEGFR-2 expression [110].
7. NOX and Cancer Drug Therapy: Implications of ROS-Mediated Cytotoxicity and Resistance
Several studies have linked ROS-derived NADPH oxidase activity with chemotherapy- or cancer
drug-induced cytotoxicity in clinical practice or under investigation (pre-clinical and clinical trial
studies). The mechanism underlying NADPH oxidase-mediated cancer drug toxicity or resistance
remains poorly understood, but interestingly, the results of several studies strongly suggest an
important role of ROS derived from NADPH oxidase activity.
7.1. ROS-Derived NADPH Oxidase Mediates Drug Cancer Cytotoxicity
Several chemotherapeutics stimulate NOX-dependent ROS production as a mechanism to mediate
cell killing [111]. Platinum-based chemotherapy is extensively used in several guideline protocols for
several clinical cancer manifestations. Platinum-derived drugs inhibit or impair thioredoxin reductase
enzyme activity, which leads to enhanced oxidative stress inside cells [112]. Paclitaxel, a
microtubule-targeting agent, increases Rac1 expression and NOX activity. An interesting result of a
previous study showed that superoxide generated in paclitaxel-treated cells was directly released
outside the cells, where it was subsequently dismutated into hydrogen peroxide, and that extracellular
hydrogen peroxide could exert cytotoxic bystander effects (mediate cytotoxicity to cells that were not
directly exposed to paclitaxel) [113].
Monoclonal antibodies and tyrosine kinase inhibitors have been largely used in clinical practice,
and these drugs improve the survival of cancer patients in comparison with conventional
chemotherapies. NOX enzymes mediate the mechanisms of action of the molecular cancer targets
obinutuzumab and tositumomab (anti-CD20 mAbs used in B-cell malignancies) and their cytotoxic
effects via the induction of superoxide derived from NOX2 [114]. NOX4 is upregulated in erlotinib-treated
head and neck cancer cells (HNSCCs), and the increased hydrogen peroxide production is correlated
with cell toxicity. HNSCCs pretreated with the anti-oxidant NAC or cells in which the NOX4 isoform
is knocked down show resistance against erlotinib-induced ROS cytotoxicity [115].
Elucidating the mechanisms underlying NADPH oxidase-mediated cell death helps to understand
the efficacy of cancer drug combination therapies. The association of doxorubicin (platin-based
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chemotherapy) and gefitinib (EGFR tyrosine kinase inhibitor) enhances hepatocarcinoma cell death
through NOX4-derived superoxides through the activation of caspase-3 [116]. However, in a
colorectal cancer cell model, an antagonistic effect between cetuximab and oxaliplatin was observed in
view of NOX1 activity. Oxaliplatin-mediated cytotoxicity was limited by the inhibition of
EGFR/NOX1-induced hydrogen peroxide production in the presence of cetuximab [117].
7.2. Overcaming Drug Resistance in View of ROS-Derived NADPH Oxidase
Overcoming drug resistance is a challenge in cancer therapy. Understanding the mechanisms related
to cancer drug resistance therapy is needed to optimize cancer treatment and design new guideline
protocols. ROS are involved in several cancer drug resistance mechanisms. P-glycoprotein (P-gp), an
ABC cassette transporter, plays a major role in chemotherapeutic resistance. Overexpressing NOX1
inhibits P-gp function and overcomes the cell resistance phenotype [118]. In breast cancer, ROS
mediate EGFR-tyrosine kinase inhibitor gefitinib resistance, and the hydrogen peroxide scavenger
catalase was able to sensitize those cells to gefitinib treatment [119]. Interestingly, the involvement of
gefitinib resistance involves signaling pathways that are redox regulated (AKT) [120]. Increased
cellular antioxidants are hypothesized as a cancer drug resistance mechanism in several models.
Histone deacetylase inhibitors are commonly used in T-cell lymphoma and have been reported to
mediate cytotoxicity in an NADPH oxidase-dependent manner. In leukemic cells, ROS derived from
NADPH oxidase increase after vorinostat and bortezomib promote cell cycle arrest, apoptosis and
DNA double-stand breaks [121–123]. In contrast, increased ROS also activate the transcription factor
Nfr2, which induces antioxidant expression. Combination therapy with vorinostat and a glutathione
depletion agent (PEITIC) reverses the drug resistance phenotype of several leukemic cells [123]. In
prostate cancer, NADPH oxidase-dependent ROS generation mediates radiotherapy resistance, and
pretreating cells with NADPH oxidase inhibitors sensitizes cells to radiation [124]. However,
modulating prostate cancer cell redox states with parthenolide, a sesquiterpene lactone, potentiates
radiotherapy-mediated cell toxicity by diminishing anti-oxidant defense via NADPH oxidase-dependent
ROS generation and activation of the PI3K/AKT/FOXO signaling pathway [125].
8. Conclusions
Advances in our understanding of the complex cellular and molecular mechanisms involved in
regulating signaling pathways related to carcinogenesis in view of ROS have provided new insight into
signaling pathway interplay and the mechanisms of action of drugs that are used in cancer treatment. In
fact, several mechanisms remain poorly understood because cancer cells may present a wide range of
phenotypes, which dictates different responses to cancer drugs.
Managing redox content inside cells is harmful. Due to the complex mechanisms involved in the
activation of NADPH oxidases, these enzymes can be targeted at several different levels of their
activity [126]. Once NADPH oxidase can regulate activities of several proteins and downstream
signaling pathways, and provides the major non-mitochondrial source of ROS inside cells, it would be
an important therapeutic target in cancer. To validate NADPH oxidase as a key player in the hallmarks
of cancer and a molecular target for cancer, it would be interesting to determine an epidemiological
profile of the expression of NADPH oxidase isoforms in cancer cells, especially those derived from
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samples of patients undergoing biopsy or surgery, and their correlation with cell signaling pathways
and therapy used for treating the disease.
Acknowledgments
Part of the work mentioned in this paper was supported by grants from the Brazilian Agencies
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de
Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo àPesquisa do Estado
do Rio de Janeiro (FAPERJ), and Instituto Nacional de Ciência e Tecnologia de Biologia Estrutural e
Bioimagem (INCTBEB).
Conflict of Interest
The authors have declared that no competing interests exist.
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Nuevas desigualdades urbanas: la apropiación global
del patrimonio en los centros históricos mexicanos
David Navarrete Escobedo*
Resumen. La desigualdad urbana en los centros patrimoniales
se ve marcada por la implantación de equipamientos turísticos,
culturales y de servicios de alta gama que generan atmósferas
aseptizadas y cosmopolitas en espacios históricamente populares y heterogéneos. Esos llamados “rescates” de los centros
históricos pueden explicarse en parte por la presencia de poblaciones extranjeras, sean residentes secundarios o turistas.
En el siglo xxi, esa presencia esconde procesos complejos que
conducen a la acentuación de las desigualdades urbanas bajo la
tutela del Estado, del mercado y de actores locales y globales. Es
una transformación económica, social, espacial y política que
excluye a los habitantes y usuarios tradicionales de los centros
históricos de las ciudades mexicanas quienes se ven despojados
del goce patrimonial.
Palabras clave. Gentrificación, turismo, patrimonio, exclusión,
ciudad media.
New urban inequalities: the global appropriation
of heritage in mexican historical centers
Abstract. Urban inequalities in heritage centres are inducted
by the creation of tourist, cultural and high-end services that
* Profesor investigador en la Universidad de Guanajuato, México. Correo electrónico:
davnav25@hotmail.com
Volumen 16, número 39, enero-abril, 2019, pp. 77-99
Andamios
77
David Navarrete Escobedo
g enerate aseptized and cosmopolitan atmospheres in historically popular and heterogeneous spaces. These so-called “rescues”
of historic centres can be explained in part by the presence of
foreign populations as secondary residents or as tourists. This
presence hides complex processes that lead the accentuation of
urban inequalities, under the tutelage of the State, the market
and local/global factors an economic, social, spatial and political transformation seeks to respond to the expectations of the
international middle classes with a higher purchasing power. In
consequence, the exclusion targets inhabitants and traditional
users who are deprived of the patrimonial enjoyment of the
historical centres in many Mexican cities.
Key words. gentrification, tourism, heritage, exclusion, middle-size city.
Introducción
La desigualdad urbana ha sido definida por David Harvey (1977) en
oposición a la justicia distributiva territorial. Ese concepto plantea que
la división de los beneficios y la asignación de cargas entre los ciudadanos sea equitativa en los procesos sociales, específicamente del trabajo
y la producción. En el siglo xxi, las desigualdades urbanas conllevan
inéditas escalas, sujetos, políticas y procesos sociales que entran en el
juego de la justicia territorial (Secchi, 2015). Trabajos como el de Sigler
y Wachsmuth (2014), han dejado en evidencia una nueva escala de la
desigualdad urbana que caracteriza a varias ciudades latinoamericanas.
Esa nueva escala es la global. Las nuevas inequidades urbanas en Latinoamérica se diferencian de las dominantes hasta el último cuarto del
siglo xx, en la medida que estas últimas se explicaban esencialmente
por condiciones endógenas de sistemas sociales, políticos y económicos que se materializaban en la realidad de un orden socioespacial más
o menos segregado en cada ciudad. Con la llegada del régimen económico neoliberal a nivel planetario, las transformaciones de las ciudades
y la desigualdad que las acompaña son interpretadas mayoritariamente
78
Andamios
Nuevas desigualdades urbanas
como procesos de adaptación de las condiciones locales a la injerencia
de actores globales come el capital transnacional, tal como lo conceptualizó la economista Saskia Sassen (1996).
En el siglo xxi, los estudios urbanos reconocen la influencia del
neoliberalismo capaz de reproducir desde lo transnacional las desigualdades estructurales de las ciudades latinoamericanas (Harvey 2013;
Secchi 2015; Paquot, 2006; Ascher, 2001, Mongin 2005). Sus puntos
de apoyo son personas, ideas, proyectos y políticas mundializadas que
se constituyen como principales vectores de la globalización y del modelo de ciudad neoliberal (Delgadillo, 2016). La movilidad es clave para
explicar el impacto que personas y capitales extranjeros con sus valores
y prácticas sociales tienen en la producción y reproducción de desigualdades urbanas en América Latina (Hayes 2018; Torres y Momsen,
2005; Augé, 1992; Urry, 2007). Esa movilidad transnacional afecta dos
actividades centrales del modelo económico de ciudad neoliberal: la de
segunda residencia y la de turismo (Hiernaux, 2012; Delgadillo, Díaz
y Salinas, 2015; Covert, 2017). Para la primera encontramos su mayor
referente en la migración transnacional con una nueva cualidad: la dirección norte-sur de las clases medias de las economías desarrolladas.
Para la segunda, encontramos su mejor manifestación en el turismo
internacional que también refuerza sus flujos masivos hacia los países
del sur desde los años setenta.
Este artículo aborda a esos sujetos transnacionales como potenciadores de las desigualdades urbanas en sectores patrimoniales de ciudades
medias del Sur Global (Lees, 2012). Es decir, indaga los procesos de
exclusión socioespacial vehiculados por la presencia importante de poblaciones extranjeras en calidad de residentes o turistas provenientes de
países del norte con altos ingresos. Desde nuestra perspectiva reconocemos, al igual que Nik, Peck y Brenner (2019), que el neoliberalismo y
las transformaciones urbanas que él induce poseen complejas interfaces
institucionales, geográficas y sociales. En ese entendido, los expatriados
se han instalado gracias a una historia de intercambios culturales y a la
interacción con élites económicas, políticas y sociales locales. Es decir,
las clases altas y medias nacionales también participan en el proceso de
desigualdad pero en menor grado, ya que incluso ellas son relegadas
por los superiores ingresos de los expatriados. Consideramos que la
Andamios
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David Navarrete Escobedo
presencia extranjera explica el nivel tan amplio de la brecha de desigualdad que la sola elite local no explicaría o no la llevaría a ese grado.
El expatriado, con el permiso de los actores locales, estaría definiendo
una parte importante de la mayor disparidad entre ricos y pobres, que
según Secchi (2009 y 2015) definen la cuestión urbana de la ciudad de
los siglos xx y xxi.
Proponemos conceptualizar las desigualdades urbanas latinoamericanas por medio de fenómenos de exclusión socioespacial presentes
en ciudades con patrimonio urbano-arquitectónico relevante que el
capital marginalizó durante buena parte del siglo xx. Esas condiciones
permitieron que por medio del proceso de Rent Gap (Smith, 2012),
se creara un nicho de oportunidad de recuperación de valor, que fue
retomado primero por las clases medias locales a las cuales se les compraron propiedades a un precio rozable en las últimas décadas del siglo
xx. Posteriormente la mayor recuperación de Rent Gap paso a manos
de extranjeros provenientes de países con mayor ingreso per cápita
que desde el siglo xxi dominan y especulan con amplios márgenes de
ganancia un mercado inmobiliario patrimonial transnacionalizado. A
principios del siglo xxi, el neoliberalismo acentuó la apropiación transnacional, residencial y de ocio al expandir un nuevo mercado global de
tierras (Sassen, 2015). El resultado determina procesos de exclusión
que caracterizan a varias ciudades medias en México, Colombia, Perú,
Ecuador o Brasil, y que no hubiesen sido posible, al menos no en esas
dimensiones, desde la base de la demanda local, metropolitana o nacional de vivienda y de turismo.
Este artículo también busca responder a dos preguntas planteadas por
el dossier de la revista Andamios: “América Latina, nuevas desigualdades
urbanas”, a saber: ¿qué papel desempeña la ciudad latinoamericana en la
generación y reproducción de las desigualdades sociales y económicas?
¿Las llamadas ciudades intermedias y pequeñas son mejores escenarios
para confrontar las inequidades urbanas, que las megaciudades? Así, a
través del análisis de los distritos históricos de Guanajuato y San Miguel
de Allende (centro-occidente de México), estudiamos nuevas geografías y
modalidades de desigualdad urbana: ciudades medias patrimonializadas,
con importantes poblaciones extranjeras, mayoritariamente estadounidenses y con usos turísticos dominantes.
80
Andamios
Nuevas desigualdades urbanas
Esta investigación de corte cualitativo aborda dos aspectos importantes que introducen nuevas desigualdades urbanas en los centros
patrimonializados: la migración por estilo de vida de jubilados ricos
y el turismo. Se analizaron los perímetros protegidos por la unesco en
Guanajuato (1988) y en San Miguel (2008) para observar su usos y las
clases sociales que las habitan/utilizan. La apropiación transnacional
de los edificios patrimoniales y del espacio, nos da cuenta de un proceso de transformación vinculado con el modelo de ciudad neoliberal
en el que el Estado se vuelve garante del capital y de actores sociales
privilegiados, élites y extranjeros preferentemente. Surge un soporte
espacial de la desigualdad urbana ensamblado de espacios habitacionales de lujo, de hoteles boutique y de boutiques de productos alta
gama, de gastronomía de autor o bistrós, de galerías de arte, así como
de espacios de alta cultura. Ese modelo polariza social y espacialmente
los centros históricos y la ciudad en su conjunto. El impacto del nuevo
uso del centro patrimonial prioritariamente para extranjeros y turistas,
se hace sentir en los indicadores de la desigualdad urbana de las ciudades patrimoniales que frecuentemente muestra ser más acentuada que
aquellas que no lo tienen (coneval, 2018).
Antiguas y nuevas desigualdades urbanas
en las ciudades latinoamericanas
La desigualdad nace con la ciudad, que a lo largo de su historia ha sido
productora y reproductora de la diferenciación entre ricos y pobres
(Paquot, 2006; Secchi, 2015). La ciudad latinoamericana no escapa a
este principio, desde las ciudades prehispánicas al periodo colonial de
los siglos xvi al xviii, la era republicana del xix y la moderna del xx; las
estructuras sociales y espaciales eran y son segregativas (De la Torre
y Navarrete, 2016; Arango, 2012; Galeano, 2014). Históricamente, las
desigualdades urbanas se han constituido de soportes materiales (equipamientos, edificios, límites, espacios cerrados y espacios urbanos) e
inmateriales (prácticas, usos, estigmas, códigos, políticas, proyectos,
valores morales y de clase) (Orbeti y Prétecille, 2016; Secchi, 2015;
Harvey, 1977). Dos mecanismos clásicos son determinantes para
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David Navarrete Escobedo
comprender las inequidades de la ciudad: la estructura de clases y la
estructura urbana. Es decir que las “formas espaciales [deben ser tratadas] no como objetos inanimados dentro de los cuales se despliegan
los procesos sociales, sino como cosas que ‘contienen’ procesos sociales en la misma medida en que los procesos sociales son espaciales”
(Harvey, 1977, p. 3). Las formas espaciales producen y reproducen las
desigualdades. Es en la mezcla del espacio con las prácticas cotidianas
de apropiación donde toda la desigualdad se concretiza (Secchi, 2013).
En el periodo neoliberal, las prácticas cotidianas de apropiación de
la estructura urbana de mayor calidad (mejor estado de conservación,
servicios urbanos eficientes y mejor equipamiento urbano) por parte
del capital privado y de clases sociales pudientes, son legitimadas por
el Estado (Delgadillo, 2016). En México, la entrada al neoliberalismo
económico cuyos detonantes fueron la privatización de bancos desde
1992; la firma de acuerdos comerciales como el Tratado de Libre Comercio de América del Norte, vigente desde 1994 y renegociado en
2018; y la concesión de recursos energéticos y minerales acelerada por
la reforma energética de 2013; implicaron desde inicios del siglo xxi el
surgimiento de inéditos ordenes desiguales en las estructuras sociales
y espaciales de las principales ciudades mexicanas (Garza, 2003). A
partir de los años ochenta, el crecimiento de las ciudades ya no dependió exclusivamente de los planes e infraestructuras financiados por
el Estado, que debilitó su intervención y fue garante para la entrada
de capitales privados con frecuencia extranjeros para la explotación y
administración de varios servicios de su exclusividad. Particularmente
la era neoliberal se manifestó con el surgimiento de un mercado inmobiliario internacionalizado, que a la par de un urbanismo de proyectos
(Carrión y Erazo, 2016), ha marcado el orden socioespacial de varias
ciudades mexicanas y latinoamericanas.
Lo global impacta en la apropiación de estructuras urbanas por parte de las clases privilegiadas en la era neoliberal de la ciudad (Sassen,
1996 y 2015). Se ha detectado a nivel planetario que con el surgimiento
de territorios emblemáticos, neoliberalismo implica polarización de las
estructuras espaciales y de clases (Secchi, 2015; Harvey, 2011; Sassen
2015; Delgadillo 2016). La ciudad neoliberal no da pruebas de heterogeneidad en sus territorios (ciudades globales o barrios I nternacionales
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de negocios, enclaves residenciales en forma de torres y clubes de alto
standing o barrios históricos tematizados por el turismo y el consumo
cultural). No hay interacción/integración con el conjunto de territorios
y grupos sociales que constituyen la ciudad neoliberal. Bajo este esquema, las desigualdades estructurales de la ciudad mexicana y latinoamericana se acentúan.
En este trabajo nos interesamos por la dimensión territorial de la
desigualdad que se encuentra estrechamente relacionada con cualidades simbólicas del territorio (Secchi, 2015; Urry, 1990). Es el caso del
valor histórico, la concentración de capital cultural, social, educativo y
de recreación de ciertos barrios patrimoniales, particularmente aquellos
reconocidos por gobiernos nacionales o por instituciones internacionales como la unesco. La apropiación de las clases altas de los espacios de
mayor calidad urbana, que incluye arquitectura, monumentos, espacios
públicos, servicios, instituciones artísticas y culturales, museos, teatros,
escuelas y fuertes tejidos asociativos, es determinante para la desigualdad territorial. Un factor determinante en este proceso es el nivel de
consumo que por el ingreso puede ser hasta nueve veces superior para
los extranjeros (Yomich, 2018) comparado con un local y en función de
él se da la exclusión. En la medida que el mayor porcentaje de riqueza
se concentra en minorías sociales, el orden socioespacial de la ciudad
neoliberal se presenta fuertemente polarizado.
El patrimonio: nuevo actor en las desigualdades urbanas
El patrimonio arquitectónico es clave para entender las nuevas desigualdades urbanas de Latinoamérica (Janoschka, Sequera y Salinas,
2014; Delgadillo, Díaz & Salinas, 2015; Sigler & Waschmouth, 2014).
En él se materializan procesos socioculturales de apropiación por parte
de poblaciones ricas como los extranjeros que emigran a ciudades del
sur (Covert, 2017; Hayes, 2018). Michaela Benson (2012) argumenta
que esa migración está determinada por imaginarios colectivos y marcos culturales de los individuos que deciden expatriarse. Ella es sensible a los valores de la clase media de los países del norte, que según
John Urry (1990) siente una fuerte nostalgia por el pasado a través de
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e xperiencias (cotidianas y turísticas) asociadas al patrimonio y a la cultura. Siguiendo a Bourdieu (1979), es con el patrimonio arquitectónico
que las clases medias occidentales buscan nutrir su capital cultural y
simbólico. En esa nueva migración se ha detectado que el patrimonio y
la cultura son ámbitos en los que los expatriados cierran el círculo de se
éxito social (Hiernaux, 2012). Esta perspectiva culturalista se combina
con los factores económicos del periodo neoliberal y las constantes
crisis financieras que por el Rent Gap hacen atractivo migrar al sur.
Además, el patrimonio construido tiene un alto valor comercial en
la organización de la ciudad neoliberal (Delgadillo, 2016). La ciudad
patrimonial se ha vuelto objeto de deseo para la inversión inmobiliaria
debido a la alta rentabilidad de los espacios de consumo turístico-comerciales (Montaner y Muxí 2012). Esos procesos han conducido a
una gentrificación patrimonial (Delgadillo, Díaz y Salinas 2015) en los
sectores centrales de las principales capitales mundiales (Londres, París,
Nueva York, Barcelona). En esos contextos se ha identificado que la
gentrificación y la turistificación producen tensiones, contradicciones y
disputas por los espacios gentrificados (Smith, 2012). Incluso que en su
proceso de acentuación, la gentrificación patrimonial participa de una
“destrucción creativa” (Harvey, 2011), pues destruye las cualidades socioespaciales que le dieron origen (Moskowitz, 2018) y puede alcanzar
una sobregentrificación (Lees, 2012), amenazando los intereses de los
actores que la crearon y que en su momento fueron los más beneficiados.
En resumen, queremos focalizar este estudio en la relación entre la
migración transnacional, el turismo y el patrimonio edificado. Es con
ese vínculo que el modelo neoliberal de ciudad orquesta transformaciones urbanas en contextos espaciales hasta hace poco excluidos del
circuito de capitales internacionales, como las ciudades medias con títulos de la unesco. Igualmente argumentamos que la concomitancia del
trinomio migración-turismo-patrimonio es determinante para la acentuación de las exclusiones territoriales. En nuestro universo de análisis,
la explotación turística del patrimonio arquitectónico se encuentra a
la par y realimenta la apropiación y la recuperación del valor de renta,
conducida dominantemente por las clases medias transnacionales.
Proponemos el surgimiento de nuevas desigualdades urbanas en
ciudades medias latinoamericanas con centros históricos de valor
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atrimonial, que desde la segunda mitad del siglo xx han comenzado
p
una apropiación liderada por migrantes extranjeros cuyo poder adquisitivo les ha permitido convertirse en los mayores propietarios o usuarios de los edificios históricos más relevantes de una ciudad para su uso
(habitacional) o de consumo (cultural, de ocio, turístico y comercial).
Nuestra modalidad de desigualdad resultaría de la relación triangular
de, primero, la migración internacional; segundo, la turistificación del
patrimonio promovida por el Estado; y tercero, el interés empresarial de
las elites económicas locales para explotar la demanda transnacional.
La desigualdad urbana se caracterizaría por la apropiación del uso de
espacios patrimoniales por dos condiciones: la primera, de un comercio de servicios de prestigio con vocación turística e internacional; la
segunda, de ocupación de habitantes extranjeros en las viviendas de los
perímetros mejor equipados y de mayor calidad estética y ambiental. El
surgimiento de una ciudad de ricos expulsaría por medio del consumo
a los pobres y aportaría una cuota de nuevos habitantes/usuarios permanentes y temporales dominada por clases medias transnacionales.
Lo anterior acentuaría las diferencias sociales. El estilo de vida de los
residentes de las clases medias transnacionales y de las prácticas turísticas, culturales y de consumo de ricos (nacionales y extranjeros),
constituyen fuerzas globales que inyectan un fuerte grado de exclusión
socioespacial que con la sola combinación de factores locales no se
daría, al menos no en esa magnitud.
Aspectos metodológicos
Los elementos patrimoniales a partir de los cuales se analizaron las
desigualdades derivadas de la turistificación y gentrificación, fueron
los edificios de mayor lujo: los hoteles boutique y los de 4 o 5 estrellas, las galerías de artes y los restaurantes tipo bistró. Se consultó
el Directorio Nacional de Unidades Económicas (dnue) del Instituto
Nacional de Estadística, Geografía e Informática (inegi) para identificar
su distribución geográfica. Se consultaron los documentos de las declaratorias de la unesco de 1988 para Guanajuato y de 2008 para San
Miguel de Allende, en los que se establecen los polígonos protegidos.
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Por último, se consultó el catálogo de monumentos históricos del Instituto Nacional de Antropología e Historia (inah) para sobreponer los
espacios gentrificados y turistificados con edificios protegidos en cada
ciudad. En el espacio público (banquetas, portales, plazas y jardines) se
realizó una observación directa para analizar las prácticas de consumo
de clases ricas.
Se identificaron los polígonos (ageb, unidades geográficas establecidas por el inegi) de uso habitacional y que coinciden con las zonas
patrimoniales en donde declararon su domicilio extranjeros. La colecta
de esos datos se realizó con los resultados del último censo disponible, el Censo General de Población del inegi 2010 y el Conteo 2015.
Igualmente se consultó el sitio Airbnb para identificar los espacios
habitacionales en renta de las ciudades. Con esos datos se analizaron
las desigualdades de la gentrificación patrimonial desde la perspectiva
habitacional en las dos ciudades.
El trabajo de campo consistió en aplicar matrices de observación sobre el edificio patrimonial y su dinámica excluyente: uso de suelo, giro,
perfil del usuario, aspectos estético-formales, cámaras de vigilancia, valet-parking, policías privadas, extensión de terrazas en espacio público,
explotación de azoteas como bares o restaurantes. Se identificó también
el uso de suelo por predio y edificio a lo largo de la calle, donde se instaló el hotel boutique y se identificaron los edificios vacantes en venta o
renta y se indagó su precio con la agencia inmobiliaria respectiva.
En el aspecto habitacional, el trabajo de campo consistió en detectar
los barrios que acusan importante presencia de vivienda de extranjeros.
Igualmente se utilizó una matriz de observación sobre calidad de la
vivienda, número de niveles, presencia de cámaras de vigilancia, electrificación, aspectos estético-formales y porosidad urbano-arquitectónica.
El trabajo de gabinete consistió en realizar los mapas temáticos de la
turistificación y la gentrificación para luego cruzarlos con datos estadísticos. Las matrices generaron un diagnostico de las modalidades de
desigualdad derivadas de la gentrificación patrimonial. Además, se realizaron entrevistas no estructuradas a informantes clave de los institutos
municipales de planeación de las dos ciudades.
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Ciudades patrimoniales desiguales:
Guanajuato y San Miguel de Allende
El perfil de la población extranjera en los centros de San Miguel de
Allende y de Guanajuato se puede dibujar con la siguiente información.
Las estadísticas disponibles para el estado de Guanajuato establecen
que se trata mayoritariamente de norteamericanos (Estados Unidos
40% y Canadá 3.5%) y sudamericanos (brasileños 4%, argentinos
3% y colombianos 2%) (inm, 2009). En cuanto al rango de edades, la
población de más de 60 años está sobrerrepresentada con un 36.5%,
le siguen la de 50 a 60 años con un 15%, en total más de la mitad de
la población tiene 50 años o más. En términos de género, 58% son
mujeres y 42% hombres. En cuanto a sus actividades principales, un
36% son jubilados, 25% trabajan, 4% estudian y el resto no especificó
su actividad. Entonces, el perfil dominante es el estadounidenses de la
tercera edad, jubilado y de altos ingresos, en promedio de tres mil 833
dólares en 2018 por hogar de retirados (Yochim, 2018).
El peso relativo de la población extranjera que daría cuenta de su
dominio en el consumo y actividades de los centros históricos es importante, sobre todo en San Miguel de Allende, donde habitan unos 17
mil extranjeros (Covert, 2017) para una población de 25 mil personas
en el polígono unesco (since, 2010). En Guanajuato, el fuerte peso relativo es del turismo (nacional y extranjero), ya que para una población
de 38 mil personas en el polígono unesco, hay una presencia anual de
cerca de un millón de turistas.
La tematización turística y la apropiación mayoritariamente extranjera del patrimonio construido es una constante en los dos casos
analizados. La primera acusa mayor grado en Guanajuato y la segunda en San Miguel. Se observa en el perímetro central de los centros
declarados como Patrimonio de la Humanidad una especialización
en usos de suelo de servicios con vocación internacional y con fuerte
presencia de usuarios extranjeros y, en menor medida, clases altas nacionales. Alrededor de los hoteles boutiques, el tejido económico se va
constituyendo de otros giros complementarios, principalmente bares
y bistrós, cafeterías, boutiques de ropa de diseñador, galerías de arte y
diseño y otros hoteles. Se ha observado en los dos casos de estudio que
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David Navarrete Escobedo
luego de la instalación de un hotel de lujo en un edificio patrimonial
se sustituyen en los inmuebles aledaños los comercios tradicionales por
otros de consumo turístico y de alta gama, igualmente en las calles y
plazas aledañas se presenta una mutación comercial. Para el caso de
San Miguel de Allende, esta especialización se observa en las calles
alrededor del Jardín Principal hacia la calle Mesones, San Francisco,
Umarán, De la Canal, Aldama y Parque Juárez. La hegemonía de los
usos de suelo turísticos, culturales y de servicio de alta gama tematizan
el centro histórico para los gustos de ocio y el alto poder adquisitivo de
las clases ricas.
Ciudad
Superficie Centro
Histórico (km2)
Nº edificios
catalogados
Nº total hoteles
Nº hoteles edificios
catalogados
Nº de turistas
anuales
Nº de Extranjeros
habitantes (inegi).
Nº boutiques
o tiendas de arte
Tabla 1. Comparación de recursos patrimoniales, turísticos y urbanos
de los centros históricos de Guanajuato y San Miguel de Allende
gto
2.22
556
747
15
936 003
303
37
sma
1.60
260
294
10
505 272
1526
53
Fuente: Elaboración propia a partir de sectur gto, 2015; dnue, 2015; since, 2010.
Cabe mencionar que en ambos centros se confirma la tendencia a la
extinción del uso habitacional en su primer cuadrante, lo que implica
ya un desplazamiento incluso de residentes adinerados. Actualmente,
la totalidad de los inmuebles patrimoniales de la hotelería de lujo y
otros servicios caros tienen uso comercial y el residencial se ha extinguido. Ello no solo en el inmueble en cuestión sino en las manzanas de
implantación por la dinámica económica que se genera en torno a un
establecimiento de esta naturaleza.
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Los usos excluyentes de los inmuebles patrimoniales conducen también una privatización del espacio urbano. Los efectos de la desigualdad urbana no terminan en el límite del inmueble de lujo, sino que se
extienden, usan, aseguran y controlan el espacio público contiguo a
su fachada o plazas. Aquí los actores políticos son determinantes pues
manifiestan un favoritismo empresarial. En el caso de Guanajuato, el
poder municipal permite el usufructo de banquetas y plazas públicas a
restaurantes, hoteles y boutiques a razón de 342 pesos mensuales por
metro cuadrado. Hacia 2014 se tenían registrados, según el servicio
local de fiscalización municipal, 1 252 m2 de uso comercial de espacio
“público” rentado a hoteles y restaurantes hacia los frentes de los inmuebles. La mayoría de esas superficies se encuentran en plazas como
la de San Fernando, la de la Paz y el Jardín Reforma.
El uso habitacional también observa una tematización turística por
la presencia de clases medias internacionales y nacionales. Se trata de la
ocupación de vivienda que a veces combina habitantes extranjeros con
visitantes también extranjeros o visitantes nacionales. Este fenómeno
queda ilustrado por la puesta en servicio de espacios de vivienda en
renta por medio de aplicaciones como Airbnb. En el caso de Guanajuato
observamos una distribución de este tipo de vivienda en los barrios de
amortiguamiento o Buffer Zone según la declaratoria de la unesco, con
una mayor concentración al norte y noreste del centro histórico. Hacia
enero de 2018, en la base de datos disponible de Airbnb, se ofertan
en Guanajuato 240 espacios de renta para turistas, de los cuales 119
son viviendas enteras, es decir, un 50% de ellas están sin habitantes
permanentes la mayoría del año. Es decir, desplazan habitantes menos
rentables para vivir cerca del centro patrimonial.
En San Miguel de Allende se repite el esquema de renta de vivienda
para visitantes en las zonas de amortiguamiento que conservan una
buena calidad urbano-patrimonial y tienen cercanía con el núcleo
turístico principal. Observamos una concentración mayor al norte del
centro histórico en las colonias Guadalupe, Azteca y Barrio del Tecolote. En enero de 2018, Airbnb registra, para el centro histórico de San
Miguel de Allende, 306 espacios de renta para turistas, de los cuales
273 son viviendas enteras que se rentan, es decir, un 90% de ellas están
sin habitantes permanentes la mayoría del año.
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En ambas ciudades, Airbnb ha surgido en colonias que no se caracterizan por tener viviendas en desuso. Por su ubicación se deduce que
la integración a la explotación se ha dado por compra a propietarios
nacionales mayoritariamente, pero comercializados por extranjeros
(Aguado, 2018). En la vivienda para uso o para renta adquirida por
extranjeros se concretiza la transferencia de capital y la recuperación
del Rent Gap a escala transnacional. Según datos de la Asociación de
Inmobiliarias de San Miguel, en 2017 se vendieron 362 propiedades
en el centro a un precio promedio de 400 mil dólares y el 65% de los
compradores eran de origen extranjero (El Universal, 2017).
Los datos del inegi sobre habitantes extranjeros en el centro de San
Miguel de Allende, muestran que se sitúan mayoritariamente en las zonas de amortiguamiento alrededor del hipercentro turistificado, coincidiendo con la zona de vivienda en renta Airbnb descrita en los párrafos
anteriores. El recorrido de campo detectó las cualidades arquitectónicas
y los usuarios presentes en esa zona, se verifica una concentración de
la población extranjera en calles y barrios bien equipados, asegurados
y con buen estado de conservación, que forman un anillo alrededor del
núcleo central de monumentos.
La cuota de habitantes de origen extranjero en el centro de Guanajuato es sustancialmente menor que la de San Miguel de Allende (Tabla
1). Ella se concentra en el polígono declarado de la unesco, aunque
en enclaves más reducidos y disgregados. Lo anterior alrededor de las
plazas y callejones de la mejor calidad urbana y paisajística de la zona
monumental. Esa distribución pasa al norte por las calles De Pocitos,
de Alonso al sur, el callejón Del Potrero (también al sur), la subida de
San Miguel, la Plaza de los Ángeles y el Callejón del Beso.
Nuevas desigualdades urbanas en ciudades medias patrimoniales
La apropiación del patrimonio arquitectónico construido aparece como
uno de los principales soportes de las nuevas desigualdades en ciudades históricas latinoamericanas. En ella se materializa una exclusión
social, económica, cultural y espacial. Así podemos establecer una
relación entre la desigualdad urbana y la mutación arquitectónica del
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patrimonio de una ciudad conducida por la explotación comercial y de
servicios requeridos para la migración por estilo de vida. El binomio de
fuerzas globales (turismo y migración) y de fuerzas locales (patrimonio,
élites políticas y empresariales), pone en cuestión el acceso al valor cultural de los edificios, tematizando los centros históricos de Guanajuato
y San Miguel de Allende. Así, la arquitectura en su calidad de capital
cultural distintivo de la clase media internacional, se adapta a funciones de consumo turísticos y de viviendas de lujo que trastocan el valor
cultural e identitario, limitando el uso para clases populares.
Al tratarse de un tipo de desigualdad urbana basada en parte en
la terciarización del patrimonio, con ello se depredan usos menos o
poco rentables como el de vivienda de clases medias y bajas locales. Se
observa en los perímetros turísticos que los dueños de las propiedades
prefieren cambiar la vocación habitacional del edificio patrimonial para
recibir rentas más altas. Así, construcciones que antes se utilizaban
como vivienda en renta, sobre todo para estudiantes y clases trabajadoras del centro, son remozadas y transformadas para fines de uso comercial. Esto aparece como una tendencia clara en el primer cuadrante de
los centros históricos analizados, en los cuales la desaparición del uso
habitacional es casi total.
Las nuevas desigualdades urbanas se activan fuertemente por las
prácticas de consumo, turísticas y culturales, de las poblaciones ricas
que ahora son dueñas del patrimonio. Parte de la economía urbana
de San Miguel esta dolarizada, específicamente en su núcleo fundacional. El consumo en restaurantes, bares, galerías de arte y hoteles
boutique, pero también de buen número de panaderías, cafeterías y
mercados “hipsterizados” es prohibitivo para el poder adquisitivo de
las clases populares de San Miguel. En ellos, una tarta puede costar 4
o 5 dólares, al igual que una cerveza artesanal local, el equivalente al
salario mínimo diario de un mexicano (88.36 pesos mexicanos). Los
productos y servicios de lujo atienden a las necesidades y gustos de
consumo prioritariamente de la población expatriada y de los turistas. Lo anterior no excluye necesariamente a las clases medias y altas
nacionales, pues ellas participan en menor medida de la exclusión
al compartir cierto cosmopolitismo selectivo. Cuando la desigualdad
urbana está mayoritariamente a cargo de un grupo extranjero, se da
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David Navarrete Escobedo
una mayor ruptura en términos sociales, culturales y económicos entre
gentrificadores y locales. En los lugares de consumo se observa una
mínima o a veces inexistente interacción entre los expatriados (que se
comunican en inglés) y los nacionales, sobre todo los de clase popular, los cuales ni siquiera frecuentan o se reconocen en los renovados
espacios patrimoniales.
Desde las declaratorias de Patrimonio de la Humanidad de Guanajuato en 1988 y de San Miguel de Allende en 2008, la presión por la
explotación turística o residencial de lujo ha ido en aumento. En estas
ciudades, diferentes niveles de gobierno, tanto municipal, estatal y federal, han participado para que la mejor parte de la ciudad pertenezca
al uso de grupos sociales privilegiados y en beneficio de sus círculos
empresariales locales aliados y de importantes capitales, sobre todo de
cadenas hoteleras. En varios proyectos se pueden rastrear conflictos de
interés en los que los compadrazgos locales o el desarrollismo se antepone al estricto cumplimiento de las normas de protección patrimonial,
al del crecimiento urbano ordenado y al del bien común o del mayor
número de ciudadanos. Podemos citar tres casos. En Guanajuato, un
hotel boutique en el barrio de la Presa (de capital nacional pero de
clientela extranjera) que durante su ampliación ha incumplido varias
normas urbanas y patrimoniales, afectando estructuras de inmuebles
vecinos catalogados. En este caso, el presidente municipal en turno
(2016) tenía vínculos familiares o profesionales con los inversionistas
hoteleros. En San Miguel, el Hotel Aqua Live (de capital extranjero)
destruyó parte de una construcción del siglo xviii (Aguado, 2016). Por
último, el fraccionamiento Puerta de Piedra en el perímetro de amortiguamiento de la declaratoria de San Miguel de Allende, por la altura
y densidad de sus edificios deterioró la calidad paisajística del centro.
Los actores políticos y económicos locales recuperan parte del Rent
Gap, pero la tendencia es que cada vez participan menos de las transacciones inmobiliarias. La recuperación del Rent Gap se dio en un primer
momento a finales del siglo xx a favor de las élites locales. Actualmente,
la mayoría de las transacciones se realizan entre extranjeros, 60% según
el Delegado de la Asociación de inmobiliarios, y los precios alcanzan
niveles muy superiores a los de las primeras adquisiciones entre locales y extranjeros (El Universal, 2017). En ese sentido, los extranjeros
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acaparan el nuevo valor de uso, las empresas extranjeras explotan los
servicios turísticos y cuando la recuperación del Rent Gap es la más alta
(después de varias transacciones inmobiliarias) ya no la comparten con
los locales.
La estructura urbana resulta importante en el proceso de constitución de las nuevas desigualdades. En ella se materializan los procesos
de disputa social y cultural de escala global. La convergencia de gentrificadores extranjeros y de turistas internacionales genera una fuerte
competencia por el espacio, esto acentúa los procesos de especulación
inmobiliaria y apuntala los precios prohibitivos que alcanzan las propiedades en los centros históricos, que pueden alcanzar precios por
30 millones de dólares para una casona en el centro de San Miguel
(Anuncio de una agencia inmobiliaria a proximidad en junio de 2018
de Hotel 11 y Jardín Juárez). En esa competencia por el espacio, las
clases pobres son las que más pierden. Ello por la mayor diferencia
de ingreso que existe entre un hogar sanmigueleño de clase media o
popular con ingreso promedio ocho mil pesos (since, 2010) y un tejano
de clase media o alta con ingreso promedio 75 mil pesos en octubre de
2018.
El sujeto extranjero aparece como el productor del espacio gentrificado (comprador-revendedor de propiedades) y el consumidor del
espacio gentrificador (usuario de servicios turísticos y habitacionales
de lujo). Lo anterior impacta en las desigualdades por medio de las
restricciones del acceso al mercado inmobiliario (especulación y dolarización) y a las prácticas de consumo (desigualdad de ingresos).
Conclusión
Las desigualdades urbanas neoliberales se suman con nuevos mecanismos a las que históricamente se han constituido en las metrópolis y las
ciudades medias de América Latina. Su construcción conceptual aquí
planteada, el desplazamiento residencial vehiculado por la presencia de
poblaciones extranjeras, se realimenta de exclusiones urbanas basadas
en prácticas de consumo, las del patrimonio histórico turistificado para
nuestros casos. Otro elemento planteado por las nuevas desigualdades
Andamios
93
David Navarrete Escobedo
neoliberales es que exclusiones propias de los centros del capitalismo
global o grandes metrópolis latinoamericanas también se manifiesten
en ciudades pequeñas, territorios que aparentemente se encuentran
fuera de los circuitos de la globalización. Con respecto a una de las
preguntas iniciales, sobre la naturaleza de la desigualdad urbana en
ciudades medias, concluimos que ellas no la contienen mejor, sino que
incluso, por su tamaño y menor diversidad de actividades económicas,
pueden ser más desiguales que las metrópolis. El modelo neoliberal
plantea la conquista de ciudades pequeñas y medias de países peri
féricos, donde ha encontrado un campo fértil para el capital global
que se beneficia de un diferencial de renta por medio de individuos
de clase media internacional o de las empresas inmobiliarias y turísticas también transnacionales, que se las apropian con ayuda de los
actores locales. En ese sentido, reconocemos que las desigualdades
urbanas asociadas a la gentrificación y a la turistificación plantean un
escenario complejo de gobiernos, autoridades, consumidores solventes
y capitalistas extranjeros y en menor medida nacionales, en un mercado inmobiliario y de servicios en el que los expatriados van ganando
terreno paulatinamente.
Los casos de San Miguel de Allende y de Guanajuato en México,
ilustran una dimensión inédita, al menos en escala cuantitativa, de una
gentrificación consumada no en favor de las clases altas y medias de la
estructura social guanajuatense (aunque participan minoritariamente),
sino en favor de clases medias transnacionales, norteamericanas principalmente, que se insertan en la cúspide social de las ciudades patrimoniales en los países del sur. Las implicaciones sociales en términos
de desigualdad por el hecho de que sean grupos extranjeros los que la
induzcan, tienen la particularidad de que la brecha económica, espacial
y cultural que se abre es mucho más amplia; ello debido al diferencial
de ingresos y a la disparidad de practicas culturales y de consumo entre
los extranjeros y las clases locales, bajas y medias e incluso altas. Esta
desigualdad cualitativa y cuantitativa no sería tan contrastada sin la
presencia de los expatriados.
Las nuevas desigualdades urbanas neoliberales se confrontan también con la necesidad de una ciudadanía transnacional o global (Borja,
2013; Paquot 2015). Desde hace al menos quince años, el prestigio de
94
Andamios
Nuevas desigualdades urbanas
San Miguel de Allende se disparó gracias a los medios de comunicación.
Los extranjeros instalados, según las consideraciones de la revista Travel
& Leisure, en la “mejor ciudad del mundo para visitar o vivir”, debido,
en parte, a su encanto cosmopolita, resienten la presión turística. Ellos
han comenzado a sentir un deterioro en la tranquilidad y calidad de
vida de sus barrios, propio de los estados avanzados de gentrificación
(Lees, 2012; Moskowitz, 2018).
Lo anterior ha llevado a que en 2018 las principales organizaciones
sociales de extranjeros como La Biblioteca de San Miguel ac, se organicen para manifestar su preocupación sobre la turistificación excesiva
del patrimonio arquitectónico de San Miguel de Allende (Aguado,
2018). La población extranjera reconoce que el factor político, sobre
todo el local, es determinante en este proceso de transformación turística que experimenta su ciudad adoptiva. Sin embargo, la Constitución
mexicana establece en su artículo 33 que los extranjeros no pueden
inmiscuirse en ningún acto político, con lo cual sus aspiraciones de
contener o manejar la presión turística, se ve truncada por su condición
de extranjería, a pesar de ser un activo social fundamental de la población del centro histórico de San Miguel de Allende. La “destrucción
creativa” de la gentrificación ha encendido su foco rojo y las disputas y
resistencias no han hecho más que comenzar.
Por último, las nuevas desigualdades urbanas latinoamericanas nos
revelan la limitación del derecho a la ciudad, que en el contexto de
las fuerzas neoliberales, sufren las poblaciones locales más vulnerables.
Los casos de Guanajuato y San Miguel, que pueden ser representativos
de varias ciudades en el subcontinente, nos muestran que es solo cuestión de tiempo para que los reacomodos socioespaciales de la ciudad
neoliberal acentúen las desigualdades territoriales. Más aún si esos procesos son impulsados por la intervención gubernamental, que busca
asegurar un nuevo desarrollo económico de la ciudad, haciendo de los
sectores patrimoniales renovados los ejes principales de las actividades
comerciales y de las clases sociales clave de la globalización.
Las importantes inversiones públicas sobre el patrimonio con fines
de atracción económica y turística benefician a los capitales extranjeros
y a una élite capitalista muy reducida. Esta especialización del territorio implica la limitación de derechos y la exclusión de actividades
Andamios
95
David Navarrete Escobedo
y de poblaciones menos adaptadas a los objetivos de competitividad
turístico-residencial internacional. En general, no hay en las intervenciones de los gobiernos locales mexicanos, una estrategia para incluir
a los ciudadanos menos acomodados, ni actividades tradicionales y
de proximidad y mucho menos de vivienda social, en los renovados
centros patrimoniales. En este contexto, la desigualdad urbana impulsada por el Estado mediante la renovación de las áreas urbanas con
declaratoria patrimonial, se presenta como una forma erosiva del derecho a la ciudad, ya que exacerba la exclusión social de una parte
de la población, especialmente en el ámbito de trabajo, de actividades
de consumo, de cultura, de vivienda y, claro está, de “su” patrimonio
histórico construido.
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Hayes, M. (2018). Gringolandia: Lifestyle Migration and the Colonial
Geographies of Late Capitalism. Minneapolis: University of Minnesota Press.
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México. Revista Homo Viator, 3(3), 23-37.
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155-171.
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mundos alternativos. Barcelona: Gustavo Gili.
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Oberti, M. y Préteceille, E. (2016). La segrégation urbaine. París: La
Découverte.
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Modulation of the Neisseria gonorrhoeae drug efflux conduit MtrE
|
Giulia Tamburrino
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English
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Spoken
| 8,565
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Modulation of the Neisseria
gonorrhoeae drug efflux conduit
MtrE Received: 6 April 2017
Accepted: 21 November 2017
Published: xx xx xxxx Received: 6 April 2017
Accepted: 21 November 2017
Published: xx xx xxxx Giulia Tamburrino1,2, Salomé Llabrés1,2, Owen N. Vickery1,2, Samantha J. Pitt3 &
Ulrich Zachariae1,2 Widespread antibiotic resistance, especially of Gram-negative bacteria, has become a severe concern
for human health. Tripartite efflux pumps are one of the major contributors to resistance in Gram-
negative pathogens, by efficiently expelling a broad spectrum of antibiotics from the organism. In
Neisseria gonorrhoeae, one of the first bacteria for which pan-resistance has been reported, the most
expressed efflux complex is MtrCDE. Here we present the electrophysiological characterisation of the
outer membrane component MtrE and the membrane fusion protein MtrC, obtained by a combination
of planar lipid bilayer recordings and in silico techniques. Our in vitro results show that MtrE can be
regulated by periplasmic binding events and that the interaction between MtrE and MtrC is sufficient
to stabilize this complex in an open state. In contrast to other efflux conduits, the open complex only
displays a slight preference for cations. The maximum conductance we obtain in the in vitro recordings
is comparable to that seen in our computational electrophysiology simulations conducted on the MtrE
crystal structure, indicating that this state may reflect a physiologically relevant open conformation of
MtrE. Our results suggest that the MtrC/E binding interface is an important modulator of MtrE function,
which could potentially be targeted by new efflux inhibitors. The introduction of antibiotics into clinical use against bacterial infections marked one of the most important
milestones in medicine. However, the high evolutionary pressure caused by the widespread use of antimicro-
bial drugs has led to the rise of antibiotic-resistant bacterial strains1. In recent decades, antimicrobial resistance
(AMR) has evolved into a major health problem, as many bacterial species have become insusceptible to a grow-
ing range of antibiotics, and we may soon face the prospect of a post-antibiotic era. Some bacteria, especially
Gram-negative organisms including strains of Neisseria gonorrhoeae, have become pan-resistant, i.e. they can
no longer be treated with any available antibiotic2. The exceptional urgency of addressing the emergence of bac-
terial multi-drug and pan-resistance has therefore been widely recognised by national and international health
authorities3 and N. www.nature.com/scientificreports www.nature.com/scientificreports www.nature.com/scientificreports Received: 6 April 2017
Accepted: 21 November 2017
Published: xx xx xxxx Modulation of the Neisseria
gonorrhoeae drug efflux conduit
MtrE (c) The narrowest site in the channel is formed by two concentric
aspartate rings comprising three D422 and D425 in the trimer (conserved across the other characterized
OMPs), which are thought to act as a selectivity gate. Viewed from the periplasmic entrance, the ring formed by
D422 is more proximal, the one from D425 more distal. Figure 1. (a) Overall structure of the trimeric MtrE efflux protein channel (PDB code: 4MT0). Each subunit
consists of a β-barrel domain, embedded in the outer membrane, an α-barrel domain, spanning ~ 100 Å
inside the periplasm and an equatorial domain, mostly unstructured, located in the middle of the α-barrel. (b) The reported crystal structure of MtrE corresponds to a conduit with a comparably large cross-sectional
area, in which the major constriction site is located at the tip of the periplasmic domain (110–120 Å from
the extracellular exit along the pore axis). (c) The narrowest site in the channel is formed by two concentric
aspartate rings comprising three D422 and D425 in the trimer (conserved across the other characterized
OMPs), which are thought to act as a selectivity gate. Viewed from the periplasmic entrance, the ring formed by
D422 is more proximal, the one from D425 more distal. the mechanisms underpinning rapid drug expulsion. For example, targeting this major driver of multidrug resist-
ance by new inhibitors may reduce the efficiency of efflux pumps and thereby restore the susceptibility of resistant
bacteria to existing antibiotics. In addition, the modulation of efflux pumps may narrow the compound spectrum
of expelled drugs and enhance the uptake of therapeutics into Gram-negative bacteria.fl p
g
p
p
g
In the case of N. gonorrhoeae, targeting efflux is of particular urgency as some of its strains have developed
pan-resistance, making efficient medical treatment by antibiotics impossible11. The most highly expressed tripar-
tite efflux pump in N. gonorrhoeae is MtrCDE12. Its outer membrane channel component, MtrE, has recently been
structurally characterised (PDB code: 4MT013). As in other RND homologues, MtrE is a homotrimeric protein
consisting of three domains: a β-barrel domain embedded in the outer membrane, an α-barrel domain, project-
ing over 100 Å into the periplasm, and an equatorial, mostly unstructured domain located in the middle of the
α-barrel (Fig. 1a). Its putative gating region is located at the periplasmic tip and is formed by two concentric rings
of conserved aspartate residues (D422 and D425; Fig. 1b,c). Modulation of the Neisseria
gonorrhoeae drug efflux conduit
MtrE gonorrhoeae has been named amongst the 12 bacteria prioritised by the WHO for accelerated
research efforts to develop new antibiotics4.fi f
Gram-negative bacteria possess a double-membrane cell envelope, which acts as a highly efficient barrier
for the inward permeation of drugs. Antibiotic agents enter these organisms predominantly via porin channels
in the outer membrane, and are often expelled directly from the periplasm, located between the two bilayers,
by active drug efflux pumps. Many highly resistant forms of Gram-negative bacteria display a combination of
single-point mutations in porin channels and upregulation of efflux pump expression5,6. Amongst these, the
major drivers of super-resistant phenotypes in Gram-negative bacteria are tripartite efflux pumps, protein com-
plexes which span both the inner and outer membrane and form a continuous aqueous pathway from the inner
membrane to the external medium7. The most clinically relevant family of Gram-negative efflux pumps is the
resistance-nodulation-cell division (RND) superfamily, consisting of three major elements: an inner membrane
pump protein (IMP), an outer membrane channel protein (OMP), and a membrane fusion protein (MFP) con-
necting the aforementioned components8. The importance of active efflux for the development of bacterial resist-
ance has been impressively demonstrated in experiments, showing that multidrug-resistance can be reversed by
knocking out the expression or inhibiting the function of efflux pumps9,10. It is therefore important to illuminate 1Computational Biology, School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK. 2Physics, School
of Science and Engineering, University of Dundee, Dundee, DD1 4NH, UK. 3School of Medicine, University of St
Andrews, St Andrews, KY16 9TF, UK. Correspondence and requests for materials should be addressed to S.J.P. (email: sjp24@st-andrews.ac.uk) or U.Z. (email: u.zachariae@dundee.ac.uk) Scientific REPorTs | 7: 17091 | DOI:10.1038/s41598-017-16995-x 1 www.nature.com/scientificreports/ Figure 1. (a) Overall structure of the trimeric MtrE efflux protein channel (PDB code: 4MT0). Each subunit
consists of a β-barrel domain, embedded in the outer membrane, an α-barrel domain, spanning ~ 100 Å
inside the periplasm and an equatorial domain, mostly unstructured, located in the middle of the α-barrel. (b) The reported crystal structure of MtrE corresponds to a conduit with a comparably large cross-sectional
area, in which the major constriction site is located at the tip of the periplasmic domain (110–120 Å from
the extracellular exit along the pore axis). Results Planar Lipid Bilayer Experiments. First we performed electrophysiological measurements, in which MtrE
was embedded into symmetric POPE planar lipid bilayers. Planar lipid bilayer electrophysiology has frequently
been used to investigate the conductance and gating of OMPs in response to membrane voltages14–17. The method
makes use of simplified membrane models, as opposed to the highly complex architecture of bacterial outer mem-
branes, which consists of asymmetric lipid bilayers, with inner leaflets containing phospholipids and outer leaflets
composed mainly of lipopolysaccharides18. It has however been shown that electrophysiology in these simplified
membrane models is a valid approach to characterise the translocation of ions across outer membrane channels19. Furthermore, the gating region of efflux outward gates is located at a large distance from the membrane in the
periplasmic space, and therefore it is expected that the simplified membrane does not substantially affect results
on the gating of these outer conduits16. g
g
It has further been demonstrated that the direction of insertion of OMPs into membranes is entirely deter-
mined by the protein structures20,21, and that OMPs spontaneously insert unidirectionally in planar lipid bilay-
ers16. In addition, MtrE possesses a large polar periplasmic domain, which is unlikely to traverse the membrane
(Fig. 1a). Since we have exclusively added the OMP to the membrane face corresponding to the periplasmic side
(cis-chamber; n = 10), it is highly probable that MtrE is unidirectionally inserted into the bilayer in our experi-
ments, despite the symmetry of the POPE leaflets. p
y
yl
When voltage-clamped at + 40 mV, the fully open state of MtrE was characterised by a unitary current ampli-
tude of 11.5 pA (Fig. 2a). MtrE was also found to open to multiple sub-conductance open states that were approx-
imately 18% (2 pA) and 60% (7 pA) of the fully open state (Fig. 2a). Addition of the binding partner MtrC to
the cis-chamber significantly increased the open probability of MtrE from 0.35 ± 0.11 to 0.86 ± 0.12 (n = 3) and
caused MtrE to gate predominantly to the fully open state (Fig. 2a). Transitions to the sub-conductance open
states of MtrE following the addition of MtrC were no longer resolved. g
g
When only a single MtrE channel was gating in the bilayer, lifetime analysis revealed that in the absence of
the MtrC membrane fusion protein MtrE displayed fast flickery gating (Fig. 2b). Modulation of the Neisseria
gonorrhoeae drug efflux conduit
MtrE So far, MtrE is the only structurally determined wild
type RND-OMP showing an open conformation in this region, expected to maximise efflux through the duct. yp
g
p
g
pfl
g
Here, we present planar lipid bilayer recordings and all-atom molecular dynamics simulations characterising
the conductance of the MtrE RND exit tunnel. Our results show that binding of the membrane fusion protein
MtrC to the external face of MtrE stabilises the open state of the channel. Our findings thus highlight the contact
region between the OMP and the MFP of MtrCDE as a switch between open and closed states of the outer drug
efflux conduit. This interface may therefore represent an attractive site for potential molecular intervention. Scientific REPorTs | 7: 17091 | DOI:10.1038/s41598-017-16995-x 2 www.nature.com/scientificreports/ Figure 2. (a) Single channel experiments reveal that MtrE can dwell between 3 different semi-open states. The
addition of the adapter protein MtrC stabilizes the most open conformation. (b) Lifetime analysis uncovers that
the effect of MtrC is to significantly reduce the closed lifetime of MtrE, so that the channel resides for shorter
intervals in the closed state. Figure 2. (a) Single channel experiments reveal that MtrE can dwell between 3 different semi-open states. The
addition of the adapter protein MtrC stabilizes the most open conformation. (b) Lifetime analysis uncovers that
the effect of MtrC is to significantly reduce the closed lifetime of MtrE, so that the channel resides for shorter
intervals in the closed state. Results Previous experiments at the cellular level
have shown that substrates cannot traverse MtrE from the extracellular space to the periplasm when complex
formation to the tripartite pump is inhibited24. This finding suggests that MtrE requires the interaction with other
pump components to induce the fully open conformation, which is expected to maximise the efficiency of efflux. In the crystal structure, the Asp gating rings (Fig. 1c) and the overall conformation of MtrE reflect an open state,
wide enough to accommodate even larger substrate molecules (Fig. 1b), although its complex binding partners
are absent. We were therefore interested if the crystallographically observed conformation corresponds to the
fully open state of MtrE.h y p
The majority of our simulations of MtrE immersed in POPC and POPE membranes show rapid closing of the
periplasmic gating region. Despite displaying complex dynamics, especially in the loop region and the outer hel-
ices, the inner helices rapidly adopt a continuously closed state on the time-scale of most of our simulations. This
finding indicated that the protonation state of the Asp gating rings may act as a potential modulator of the closing
transition25. We therefore tested if a different protonation state of the Asp side chains affected channel closing. As
shown in Supplementary Fig. S1, protonation of the Asp gating rings slightly lowers the propensity for closing,
however a consistently stable open state of the gating region is still not observed. This observation suggests that
additional factors are likely to play a role in stabilising the open state of the channel. Our experimental data show
that binding of MtrC leads to the stabilisation of MtrE in an open conformation for substantial lengths of time. Taking these results together, we suggest that binding partners are required to retain MtrE in a fully open confor-
mation, while protonation changes within the gating region, possibly induced by binding, may further stabilise
the open state. In addition, our results indicate that crystal contacts may be able to at least partially compensate
for the lack of interactions with other pump components to retain MtrE in the open state. In our single channel
experiments, we observed MtrE to adopt a mostly closed state in the absence of MtrC, while the pH of the solu-
tion was not altered, emphasising the major role of the MtrE/MtrC contacts which we find for gating the channel. Results The addition of MtrC to the
periplasmic face of MtrE altered channel gating causing the channel to dwell for longer sojourns in the fully open
state, demonstrated by an increase in the apparent open time from 1 ms to 10.5 ms (Fig. 2b). This suggests that
MtrC alters the conformation of the MtrE protein and stabilizes the channel in the fully open state. In 3 out of
13 experiments, MtrE gated predominantly to the fully open state even in the absence of MtrC adapter protein,
suggesting that once in this state, channel openings are stabilised. Construction of a current-voltage relationship
for the fully open state of MtrE displayed an open channel conductance of 304 ± 7 pS (Fig. 3a).t y
y
g
Under non-symmetrical conditions (trans 210 mM KCl:cis 460 mM KCl), the reversal potential shifted to
−5.5 ± 0.8 mV (Fig. 3b), revealing that MtrE is approximately 1.7 fold more permeable for K+ than for Cl−. This
characterises MtrE as a slightly cation selective channel. The best-studied OMP homologues, TolC from E. coli
and OprM from P. aeruginosa, also display a preference for cations, however to a much larger degree. For instance, Scientific REPorTs | 7: 17091 | DOI:10.1038/s41598-017-16995-x 3 www.nature.com/scientificreports/ Figure 3. (a) The I/V plot of the MtrC/E protein complex is linear in the experimental voltage range (between
−40 and +40 mV) and the conductance for the fully open state is 304 ± 7 pS. (b) The reversal potential of
MtrE in non-symmetrical conditions (trans 210 mM KCl:cis 460 mM KCl), indicated by a dotted line, is at
−5.5 ± 0.8 mV revealing slight cation selectivity. Figure 3. (a) The I/V plot of the MtrC/E protein complex is linear in the experimental voltage range (between
−40 and +40 mV) and the conductance for the fully open state is 304 ± 7 pS. (b) The reversal potential of
MtrE in non-symmetrical conditions (trans 210 mM KCl:cis 460 mM KCl), indicated by a dotted line, is at
−5.5 ± 0.8 mV revealing slight cation selectivity. TolC has been found to be 16.5 fold more permeable for K+ than for Cl− 22, and the selectivity of OprM has been
stimated to be similar to that of TolC23. MtrE Dynamics and Computational Electrophysiology. Scientific REPorTs | 7: 17091 | DOI:10.1038/s41598-017-16995-x Results (b, top) The density map of K+ (contoured at 2% SD, in red) and Cl− (contoured at 5% SD, in blue)
highlights the influence of this highly basic region on ion flow through the channel. therefore restrained the overall backbone conformation of the protein and investigated by computational elec-
trophysiology simulations (CompEL) if the pore geometry captured in the crystal can explain the experimentally
observed channel currents. Due to the limited time-scale of the simulations, we used slightly raised transmem-
brane voltages compared to our experimental conditions, in order to drive ion permeation and ensure sufficient
sampling26. Figure 4 shows that the current we obtain for single open MtrE conduits is generally in good agree-
ment with our experimental data. The estimated maximum conductance of the open pore is 324 ± 34 pS, which
is similar to the maximum experimental conductance of 304 ± 7 pS. In the simulations, we observe ion selectivity
ratios between 1:1.2 ± 0.5 (K+:Cl−, at 100 mV) and 1:1.3 ± 0.5 (at −100 mV). These voltages were chosen as they
are close to the experimental range of voltages, while at the same time allowing us to record sufficient sampling
of ion permeation for robust selectivity estimates. The selectivity values are in generally good agreement with the
experimental ion selectivity, although Cl− ions show a slightly higher permeability in our simulations compared
to the experiments. p
We modelled an additional MtrE conformation, based on the most dilated state observed for TolC so far,
where it is in complex with the complete RND machinery MacAB-TolC27 (Supplementary Fig. S2). For this
model, we measure an in silico conductance of 597 ± 70 pS, which is considerably higher than our experimen-
tal conductance. Our data therefore suggests that the crystal structure of MtrE is a good representation of the
overall conformation of the fully open state of this conduit. Our results further show that protein-protein con-
tacts between MtrE and MtrC stabilise the fully open pore in the electrophysiological experiments, whereas in
isolation, such as in our computer simulations, the open conformation of MtrE is likely to undergo rapid transi-
tions to more closed states unless restrained. Results d
h
l
f
l bl
h
h
d
l
f g
ji
g
g
To date, there is no experimental information available on the three-dimensional structure of MtrC,
its mode and position of binding to MtrE. To mimic the stabilising effect of the membrane fusion partner 4 www.nature.com/scientificreports/ Figure 4. (a) Red dots display the experimental current-voltage relationship in symmetrical 210 mM KCl
solution, black dots show the current/voltage relationship obtained from the MD simulations near the
experimental voltage range. Voltages of ±0.05, ±0.10 and ±0.25 V were applied along the z-axis using external
electric fields as implemented in GROMACS57, together with the charmm36 force field and positional restraints
of 200 kJ/mol · nm2 on MtrE heavy atoms as detailed in the text. We observe excellent agreement between the
experimental and the computational currents. The computational x-axis and the experimental x and y-axis
error bars are smaller than the data points. (b, bottom) Sequence alignment of the three electrophysiologically
characterised efflux OMPs. MtrE shows an abundance of arginine residues in the periplasmic loop region in
addition to a series of aspartate residues lining the interior of the pore, which is not present in homologous
proteins. (b, top) The density map of K+ (contoured at 2% SD, in red) and Cl− (contoured at 5% SD, in blue)
highlights the influence of this highly basic region on ion flow through the channel. Figure 4. (a) Red dots display the experimental current-voltage relationship in symmetrical 210 mM KCl
solution, black dots show the current/voltage relationship obtained from the MD simulations near the
experimental voltage range. Voltages of ±0.05, ±0.10 and ±0.25 V were applied along the z-axis using external
electric fields as implemented in GROMACS57, together with the charmm36 force field and positional restraints
of 200 kJ/mol · nm2 on MtrE heavy atoms as detailed in the text. We observe excellent agreement between the
experimental and the computational currents. The computational x-axis and the experimental x and y-axis
error bars are smaller than the data points. (b, bottom) Sequence alignment of the three electrophysiologically
characterised efflux OMPs. MtrE shows an abundance of arginine residues in the periplasmic loop region in
addition to a series of aspartate residues lining the interior of the pore, which is not present in homologous
proteins. Results Supporting this notion further, we constructed a molecular model
of the bound state of MtrE and MtrC, based on the homologous complex of AcrAB-TolC resolved by electron
cryo-microscopy, which exhibits a similar degree of opening of the OMP gating region as the MtrE crystal struc-
ture8. Simulations of this complexed model indicate an increased tendency to remain in an open state without
undergoing further protonation changes (Fig. S3). g
g
p
g
g
Our analysis of ion trajectories in the simulations of MtrE under voltage (Fig. 5) shows that, although both
anions and cations generally occupy most parts of the MtrE channel lumen to similar extent, there are regions
in which important differences are observed. Especially the periplasmic gating region, but also sections of the
transmembrane β-barrel show a substantially reduced K+ density, particularly evident at negative membrane
potentials (Fig. 5a,b, left). A superposition of the positions of ions along the trajectories shows the constrictions
observed for the cation pathway at the periplasmic entrance, highlighting the importance of this gating region,
which includes a high density of arginine residues (see Fig. 4), for the control of ion conduction. Furthermore, we
observe that the channel lumen and gating region are well-hydrated in the open state of MtrE (Fig. 5a,b, right). Dewetting transitions play a major role in the gating process of many membrane channels28,29. To ascertain if
dewetting plays a part in the closing of MtrE, we further analysed the simulations of MtrE, in which the channel
rapidly adopted a closed state (Fig. S1). We found that even when the gating region shows a minimal level of
openness, occupation with water and a continuous water chain between the extracellular and periplasmic side
are still observed (Fig. S1B). Our findings suggest that dewetting of MtrE is unlikely to underpin the gating of this
channel for ion conduction. Scientific REPorTs | 7: 17091 | DOI:10.1038/s41598-017-16995-x 5 www.nature.com/scientificreports/ Figure 5. (a,b) Pathways of K+ and Cl− ions in the MtrE channel (depicted as ion density) and average water
density in the channel during simulations at (a) +100 mV and (b) −100 mV. The panels show the extracellular
side of MtrE at the top and the periplasmic gate at the bottom of each figure. Under both conditions, a thinning
out of the ion density at the periplasmic gate can be observed, especially for K+ ions. Results (c) Superposition of all
K+ (orange) and Cl− (green) ions during 250 ns (1 frame every 2.5 ns, 100 total frames) at +100 mV clearly
shows this constriction of the ion pathways within the gating region, representing the major bottleneck for ion
permeation. Figure 5. (a,b) Pathways of K+ and Cl− ions in the MtrE channel (depicted as ion density) and average water
density in the channel during simulations at (a) +100 mV and (b) −100 mV. The panels show the extracellular
side of MtrE at the top and the periplasmic gate at the bottom of each figure. Under both conditions, a thinning
out of the ion density at the periplasmic gate can be observed, especially for K+ ions. (c) Superposition of all
K+ (orange) and Cl− (green) ions during 250 ns (1 frame every 2.5 ns, 100 total frames) at +100 mV clearly
shows this constriction of the ion pathways within the gating region, representing the major bottleneck for ion
permeation. www.nature.com/scientificreports/ area of about 30–40 Å2 and continuous hydration of the pore (Fig. S1). As opposed to TolC, which shows a strong
preference for cations22, the cation selectivity we observe for MtrE is about 10-fold smaller. In the simulations, a
region of diminished cation density in the channel coincides with an abundance of arginine residues in the loop
region at the periplasmic tip of MtrE, which precedes the channel-internal conserved aspartate gating rings from
the periplasmic side, suggesting a role for these arginine groups in determining the ion selectivity of MtrE (Figs 4
and 5). Ultimately however, additional experiments will be required to unequivocally identify the determinants
of the lowered level of MtrE ion selectivity. Notably, the major outer membrane porins of Neisseria, PorA and
PorB, and of related organisms (e.g. Omp32 from Delftia acidovorans), show strong anion selectivity33–35 in con-
trast to the cation-selective porins of other Gram-negative organisms (such as OmpF from E. coli)36. The reduced
cation-selectivity of N. gonorrhoeae MtrE could thus be linked to an increased inward permeability of the outer
membrane of Neisseria species for anions compared to other Gram-negative bacteria.i p
p
g
All of these previous findings, together with our results, suggest that the inward and outward permeability
of neisserial outer membranes differs substantially from that of model organisms, frequently used to investigate
the determinants of Gram-negative cell wall permeation, such as E. coli. In particular, the uptake and efflux of
antibiotics may be underpinned by different principles. According to our data, the effect of different model lipids
on MtrE function is rather small, but the complex membrane composition of neisserial outer membranes may
add a further layer of modulation and control of neisserial OMPs, which may differ from observations made in
other Gram-negative bacteria. g
The differences to permeation across E. coli TolC, for example, may have important consequences for the
design of antibacterials against these difficult-to-treat Gram-negative pathogens. In particular, N. gonorrhoeae
strains are amongst the most antibiotic-resistant bacteria, displaying pan-resistance against all presently available
antibiotic agents4,11. This confers a high degree of urgency to the development of novel therapies against N. gon-
orrhoeae infections.fl Active drug efflux of a broad spectrum of antimicrobials is one of the major factors driving the development of
resistance in Gram-negative bacteria2. Methods
C
t t Computational methods. The 3.29-Å resolution crystal structure of MtrE (PDB code: 4MT0)13 was used
as a starting structure. The sulfate ion was removed and the N- and C-terminal residues were capped using
acetyl and N-methyl amide groups, respectively. We note that there is relatively weak electron density for resi-
dues 203–212 in the crystal structure. All atomistic molecular dynamics simulations were performed using the
GROMACS-5.1.1 software package39. We tested the robustness of our results by using different forcefields. tfi
First, the protein was embedded in pre-equilibrated and solvated POPC (1-palmitoyl 2-oleoyl sn-glycero
3-phosphatidyl choline) bilayers containing 288 lipid molecules (177 after insertion of the protein) using the
GROMACS g_membed utility40,41. The system was solvated and K+ and Cl− ions were added to neutralise the
system and to reach a concentration of 200 mM. Here, the Amber ff99SB-ILDN force field was used for the pro-
tein42,43, and Berger parameters adapted for use within the Amber ff99SB-ILDN force field were employed for
the lipids44,45. The SPC/E model was used for the waters46 and Joung/Cheatham III parameters were employed
for the ions47. The systems were minimised and then equilibrated with position restraints on protein heavy
atoms of 1000 kJ/mol · nm2 for 20 ns. Water bond angles and distances were constrained by SETTLE48, while all
other bonds were constrained using the LINCS method49. The temperature was kept constant at 310 K, using the
v-rescale method50 with a time constant of 0.2 ps. The pressure was kept constant throughout the simulations
at 1 bar, using a Berendsen barostat51 with semi-isotropic coupling. The application of the virtual site model for
hydrogen atoms52 allowed the use of a 5-fs time step during the simulations. For the CompEL simulations, the
protocol described by Kutzner et al.26 was used; the system was duplicated along the z axis to construct a dou-
ble bilayer system, and ionic imbalances from 2 to 6 Cl− ions were used between the aqueous compartments to
generate a range of transmembrane potentials from ~−500 to ~500 mV. Some simulations made use of protein
backbone heavy atom position restraints of 200 kJ/mol · nm2 or 1000 kJ/mol · nm2 as indicated in the text.i y
p
Furthermore, in additional sets of simulations, we employed the CHARMM36 force field for the protein,
lipids and ions53. For water molecules, the TIP3 model was used54. www.nature.com/scientificreports/ Our results suggest that the less stringent selectivity for cations in MtrE we
found here may contribute to the efficiency with which a very wide range of antibiotics is expelled from N. gonor-
rhoeae by the MtrCDE efflux system. The diminished cation-selectivity may however also allow the exploration
of new chemical space for the design of clinically usable efflux inhibitors. Most previous attempts at designing
efflux pump inhibitors have failed in clinical studies due to toxicity problems related to the cationic pharma-
cophore, which is required for efficient competitive inhibition of the pumps37. Some inhibitors were also targeted
against the internal Asp gating rings of the OMP, but clinical success was not achieved by these cationic inhibitors
either38. The different electrostatics of the MtrE interior, and its reduced preference for cations, may enable drug
researchers to design novel inhibitors with different charge properties.fl gf
g p
p
Importantly however, our results indicate that it may not be necessary to block the outer gate of the efflux
system through orthosterically binding inhibitors. We show that the opening of MtrE is regulated by allosteric
binding events of the adapter protein MtrC on the periplasmic outer face of the pore. Association with MtrC
alone is sufficient to keep MtrE in a prolonged open state. The binding of MtrC may be linked to further proto-
nation changes at the interface and pore interior of MtrE. These findings suggest that the MtrE-MtrC binding
interface may be an attractive targeting site for the development of allosterically acting efflux inhibitors. These
inhibitors would no longer compete for the orthosteric drug binding sites in the efflux pumps, but still regulate
the openness of, and thereby the efficiency of drug expulsion across, the outward conduit. This new modulation
mechanism may potentially facilitate the design of a new type of inhibitor, avoiding previous chemotypes known
for their toxicity. Discussion Both our experimental data, obtained from single-channel electrophysiology, and our computational results
suggest that the channel MtrE is mainly present in a closed state when it is unbound from the rest of the efflux
machinery. These findings are in agreement with electrophysiological recordings on homologous OMPs16,30, and
biochemical data on MtrE31,32. Our results show that the interaction of the adapter protein MtrC with MtrE is
sufficient to stabilise the complex in an open state, which conducts large ion currents in accordance with the
cross-section of the channel observed in the MtrE crystal structure13. This is, to our knowledge, the first time that
the fully open state of an OMP from an efflux pump could be characterised without inserting mutations in the
gating region, as previously reported, e.g., for TolC from E. coli30. g
g
g
p
y
p
g
We attribute the subconductance states we observe in the electrophysiological recordings to the range of
closed conformations we find in the MD simulations, which show a remaining open channel cross-sectional Scientific REPorTs | 7: 17091 | DOI:10.1038/s41598-017-16995-x 6 www.nature.com/scientificreports/ Scientific REPorTs | 7: 17091 | DOI:10.1038/s41598-017-16995-x www.nature.com/scientificreports/ simulations at 1 bar, using a Parrinello-Rahman barostat56 with semi-isotropic coupling. Here, a 2-fs time step was
used, and the proteins were inserted in a POPE (1-palmitoyl 2-oleoyl sn-glycero 3-phosphatidyl ethanolamine)
membrane. A constant electric field was applied to generate membrane potentials from ~−250 to ~250 mV57. The systems were constructed by using the CHARMM-GUI webserver58. In the production simulations, protein
backbone heavy atom position restraints of 200 kJ/mol · nm2 were applied. y
p
pp
Structural alignment was performed using the Jalview suite59, together with the Clustal W program60. Homology modelling was performed with the MODELLER9.16 suite61,62, using the dilated Cryo-EM structure of
TolC27 (Fig. S2). During the modelling process, symmetry restraints were applied on MtrE Cα atoms, to take into
account the threefold symmetry of the protein. Protein production. The MtrE protein was produced based on a previous protocol by Lei et al.13, but using
the different expression plasmid pET28a. It was solubilised from the membrane in n-dodecyl-β-D-maltoside
(DDM). The MtrC protein was produced based on a previous protocol by Janganan et al.32, increasing the growth
period to 18 hours to yield a sufficient amount of protein. Analytical gel filtration suggested that the protein runs
in several multimers, mainly dimers and hexamers, which correlates well with the protocol used32. The purity of
the samples was confirmed by SDS-PAGE, and their identity was confirmed through peptide mass fingerprint-
ing (Fig. S7), conducted by the FingerPrints Proteomics Facility of the University of Dundee. Conductance measurements and analysis. Current recordings were monitored under voltage-clamp
conditions using a BC-525C amplifier (Warner Instruments, Harvard) following established methods14,63. Recordings were low-pass filtered at 10 kHz with a 4-pole Bessel filter, digitized at 100 kHz using a NIDAQ-MX
acquisition interface (National Instruments, Texas, USA). Data were recorded to a computer hard drive using
WinEDR 3.6.4 (Strathclyde University, Glasgow, UK). The recordings were subsequently filtered at 800 Hz(−3 dB)
using a low-pass digital filter implemented in WinEDR 3.6.4. Experiments were performed at room temperature
(20–22 °C). The MtrE protein was added to the cis chamber and incorporations made at +40 mV, under contin-
uous stirring. Unless otherwise stated, single-channel events were recorded in symmetrical 210 mM KCl. For
selectivity measurements, the cis-chamber contained 510 mM KCl and the trans-chamber contained 210 mM KCl. Selectivity was computed using the Goldman-Hodgkin-Katz equation. www.nature.com/scientificreports/ Erev was taken as the voltage at which no
current flow was detected and was corrected for junction potential. Lifetime analysis was performed using TAC
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t t The temperature was kept constant at 310 K,
using the Nose-Hoover method55 with a time constant of 0.1 ps. The pressure was kept constant throughout the Scientific REPorTs | 7: 17091 | DOI:10.1038/s41598-017-16995-x 7 www.nature.com/scientificreports/ Scientific REPorTs | 7: 17091 | DOI:10.1038/s41598-017-16995-x www.nature.com/scientificreports/ www.nature.com/scientificreports/ 22. Andersen, C., Koronakis, E., Hughes, C. & Koronakis, V. An aspartate ring at the TolC tunnel entrance determines ion selectivity and
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We thank Sharon Shepherd, Thomas Eadsforth and the Protein Production Unit of the University of Dundee
for expression and purification of the MtrC and MtrE proteins. We are grateful to Gavin Robertson and Ben
Reilly O’Donnell for assistance with the electrophysiology measurements. We acknowledge funding through
the Wellcome Trust Interdisciplinary Research Funds (grant WT097818MF), the Scottish Universities’ Physics
Alliance (SUPA), Tenovus Tayside (grant T16/30) and the Tayside Charitable Trust. O.N.V. has been funded
through a BBSRC CASE award (BB/J013072/1). Author Contributions S.L., S.J.P. and U.Z. conceived and designed the research, G.T. conducted the research, all authors analysed the
data, G.T., S.J.P. and U.Z. wrote the manuscript, and all authors edited and reviewed the manuscript. Additional Information Additional Information
Supplementary information accompanies this paper at https://doi.org/10.1038/s41598-017-16995-x. Supplementary information accompanies this paper at https://doi.org/10.1038/s41598-017-16995-x. Supplementary information accompanies this paper at https://doi.org/10.1038/s41598-017-16995-x. Competing Interests: The authors declare that they have no competing interests. Competing Interests: The authors declare that they have no competing interests. p
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mitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the
copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Scientific REPorTs | 7: 17091 | DOI:10.1038/s41598-017-16995-x www.nature.com/scientificreports/ Imaging α-Hemolysin with Molecular Dynamics: Ionic Conductance, Osmotic Permeability, and
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Elevated expression of LRRC15 Shows Putative Tumor-promoting Characteristics in tumor microenvironment of breast cancer
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Elevated expression of LRRC15 Shows Putative
Tumor-promoting Characteristics in tumor
microenvironment of breast cancer
Lei Li
Nanjing Medical University
Yin-Jiao Fei
Nanjing Medical University
Ming-Xing Liang
Nanjing Medical University
Hong-Lei Zhou
Nanjing Medical University
Guan-Qun Wo
Nanjing Medical University
Xin-Yi Shao
Nanjing Medical University
Jian-Yun Lan
Nanjing Medical University
Wen-Xiu Xu
Nanjing Medical University
jinhai tang ( jhtang@njmu.edu.cn )
Jiangsu province hospital https://orcid.org/0000-0003-2016-4216
Primary research
Keywords: LRRC15, breast cancer, bioinformatic analysis, tumor-promoting, tumor microenvironment
Posted Date: November 9th, 2021
DOI: https://doi.org/10.21203/rs.3.rs-871638/v1
License: This work is licensed under a Creative Commons Attribution 4.0 International License.
Read Full License
Page 1/23
Abstract
Background
Leucine-rich repeat containing 15 (LRRC15), belongs to the LRR superfamily and has emerged as a
marker of cancer-associated fibroblasts. It was found to be particularly upregulated in breast cancer
(BCa). This study aimed to investigate the correlation between LRRC15 expression and immune
microenviroment and visualize its prognostic landscape in BCa.
Methods
The mRNA expression level, prognostic value, correlation of immunity, gene-gene interaction network of
LRRC15 in BCa were analyzed utilizing the Oncomine, Gene Expression Profiling Interactive Analysis
(GEPIA), UALCAN, Kaplan-Meier plotter, and TIMER database. We next analyzed the biological functions
of LRRC15 and pathways of its co-expressed genes, and its correlation with immune system responses
via the Metascape and GeneMANIA database, respectively. We validated the expression of LRRC15 in
BCa via western blot and IHC assays and analyzed its correlation with clinicopathological parameters.
Results
We explored LRRC15 expression in multiple types of cancer based on the Cancer Genome Atlas (TCGA)
database, with the effect being particularly pronounced in BCa. Both mRNA and protein abundance of
LRRC15 were significantly elevated in BCa as compared to its non-tumor counterparts. Overexpression of
LRRC15 significantly associated with reduced overall survival. LRRC15 knockdown significantly inhibited
cell proliferation and cell cycle in BCa cells. There were significant positive correlations between LRRC15
expression and tumor-infiltrating immune cells (TIICs), with a particularly strong effect on macrophage
infiltration. Moreover, markers of TIICs exhibited different LRRC15-related immune infiltration patterns.
GSEA analysis showed that upregulated expression of LRRC15 was related to ECM receptor interaction,
focal adhesion, regulation of actin cytoskeleton, and TGF Beta signaling pathway.
Conclusions
These findings revealed that LRRC15 served as a novel prognostic biomarker and putative oncogene for
BCa by promoting cell proliferation, giving a novel hint for therapeutics of BCa.
Introduction
Breast cancer (BCa) is the malignancy with the highest prevalence and has the second highest cancerrelated mortality rate in females worldwide [1]. BCa is a heterogeneous cancer consisting of different
molecular subtypes, differs in terms of tumor biology, incidence, risk of cancer progression, response to
Page 2/23
diverse therapies, clinical outcomes, and sites of metastases [2, 3]. Treatments for BCa range from
surgery to collaborative combinations of surgery, chemotherapy, endocrine therapy, targeted therapy and
radiation therapy [4, 5]. Simultaneously, despite explosive therapeutic advances, BCa exerts a substantial
impact on the healthcare system and the statistics of mortality and morbidity are daunting [1, 6].
Therefore, it is urgent to identify the crucial molecular events related to the malignant transformation of
BCa cells, providing a theoretical basis for the early diagnosis, monitoring progression, and molecular
therapy of BCa.
Leucine-rich repeat containing 15 (LRRC15), a 581 amino acid type I transmembrane protein, belongs to
the LRR superfamily and has emerged as a marker of cancer-associated fibroblasts (CAFs) recently [7].
Remarkably, LRRC15 was highly expressed on stromal fibroblasts in multiple solid tumors, such as
breast, head and neck, lung and pancreas, while was lowly expressed in most normal tissues [7, 8]. The
LRR family generally regulates a broad repertoire of functions, including signal transduction, cell
adhesion, DNA repair, and RNA processing [9]. Augmented levels of the LRRC15+ CAF signature was
linked with poor response to immune checkpoint blockade therapy, making it a determinant of patient
response to cancer immunotherapy [10]. Since only a fraction of patients exhibit durable responses to
cancer immunotherapy [11, 12], it is pivotal for us to have a broader view of the entire tumor
microenvironment (TME) and seek potential targets.
Here, we sought to determine the expression of LRRC15 and its correlation with TME of BCa. Moreover,
we explored the tumorigenic roles of LRRC15 by functional assays in vitro and revealed the critical links
between LRRC15 and tumor growth in BCa. Our results offered novel insights into the functional role of
LRRC15 in BCa, thereby highlighting a potential mechanistic basis whereby LRRC15 influenced tumor
evolution and progression in TME of BCa.
Materials And Methods
Raw Data acquisition and processing
Transcriptome RNA-seq data of normal and cancer tissues in pan-cancer were extracted from TCGA (The
Cancer Genome Atlas) database (https://portal.gdc.cancer.gov/). TCGA research network is a publicly
funded project designed to to catalog and explore major genomic alterations with the purpose of
establishing an integrated 'atlas' of cancer genomic profiles [13, 14].
Oncomine database Analysis
Oncomine (www.oncomine.org), a publicly accessible online database and web-based data-mining
platform, deliver plentiful cancer microarray information and genome-wide expression analysis [15]. This
cancer microarray database was applied to investigate the transcriptional levels of LRRC15 in various
clinical cancer specimens compared with those in normal tissues.
UALCAN database analysis
Page 3/23
UALCAN (http://ualcan.path.uab.edu) web-portal is an omnibus and interactive database based on TCGA
RNA-seq and clinical data of multiple cancer types [16]. We utilized this platform to elaborate the distinct
expression levels of tumor and normal tissues in subtypes of BCa.
Gene Expression Profiling Interactive Analysis (GEPIA)
database analysis
GEPIA (http://gepia.cancer-pku.cn/index.html) is a recently developed web server utilizing a standard
processing pipeline and consist of 9,736 tumors and 8,587 normal samples data from TCGA and the
Genotype-Tissue Expression (GTEx) projects [17]. It provides fast and customizable functionalities such
as differential expression analysis, similar gene detection, patient survival analysis, profiling according to
cancer types or pathological stages, correlation analysis, and dimensionality reduction analysis.
Kaplan-Meier plotter database analysis
Kaplan-Meier plotter (http://kmplot.com/analysis/) is a platform providing information about the
correlation of gene expression level with the survival of cancer patients, including BCa, liver cancer,
gastric cancer, ovarian cancer, and lung cancer [18]. Information regarding HRs, 95% CIs and P-values can
be accessed at the webpage.
Human Protein Atlas (HPA) database analysis
The Human Protein Altas (HPA) (https://www.proteinatlas.org/) is a public web interface that can be
utilized to identify the distribution and expression of target proteins. It uses special antibodies and
contains immunohistochemical expression data for 20 kinds of tumors [19]. In this study, we confirmed
prognostic significance and compared the protein expression levels of LRRC15 between normal and BCa
tissues.
TIMER (Tumor Immune Estimation Resource) database
analysis
TIMER (https://cistrome.shinyapps.io/timer/) is a user-friendly web interface designed for analyzing
immune cell infiltrates in diverse malignancies [20]. LRRC15 was selected to investigate via “Gene
module” and generated scatterplots to visualize the relationship of its expression with immune infiltration
level in BCa.
GeneMANIA database analysis
GeneMANIA algorithm (http://genemania.org/), an intuitive web interface, was applied to establish a
biological network for LRRC15 in terms of physical interaction, prediction, co-expression, colocalization,
and shared protein domains, and evaluate the functions of network components [21]. A figure where
nodes symbolize genes and links represent networks was used to display interactions.
Functional And Pathway Enrichment Analyses
Page 4/23
Genemania database has been applied to provide information regarding the predicted interaction of
LRRC15. TCGA breast cancer samples were grouped as high LRRC15 expression group and low LRRC15
expression group based on the median transcriptional expression of LRRC15. Gene set enrichment
analysis (GSEA) software was applied to evaluate potentially biological mechanism associated with
LRRC15 mRNA expression levels. A nominal P-value <0.05 and FDR <0.25 were considered to a
statistically significant gene set.
Cell lines, culture and reagents
A panel of BCa cell lines including MDA-MB-453, MDA-MB-468, BT-549, HS 578T, MCF7, T47D, ZR-75-1,
SUM1315MO2, HCC-1806, SK-BR-3, MAD-MB-231, BT-474 and human normal breast epithelial cells (MCF10A) were obtained from Cell Bank of Type Culture Collection, Chinese Academy of Sciences (Shanghai,
China). MAD-MB-231, T47D, SUM1315MO2, SKBR-3, HS-578T lines were cultured in DMEM
medium(KGM12800-500), BT-474, ZR-75-1, HCC-1806, BT-549 lines were cultured in 1640 medium
(KGM31800N-500), MDA-MB-453, MDA-MB-468 lines were cultured in DMEM medium (KGM, KGM41300500), MCF7(KGM12800-500,Insulin), MCF-10A (LONZA MEGM Bullet Kit(CC-3151 & CC-4136)),
supplemented with 10% heat-inactivated fetal bovine serum (FBS, Gibco).
Antibodies and reagents
Primary antibodies for Western blotting and immunohistochemical staining included anti-LRRC15
(Thermo Fisher Scientific, PA5-57298), anti-p63 (MXB Biotechoiogles, MAB-0694), SMA (MXB
Biotechoiogles, MAB-0890), Calponin (MXB Biotechoiogles MAB-0712), Ki-67 (MXB Biotechoiogles, MAB0672).
Tissue specimens and Immunohistochemistry (IHC)
analysis
The immunohistochemistry protein expression was archived from formalin-fixed paraffin-embedded
(FFPE) tumors from patients who underwent surgical treatment between February 2019 and February
2020 at Yancheng NO’1 People’s Hospital. This study was approved by the Yancheng NO’1 People’s
Hospital Review Board. Written informed consent was obtained from these patients.
FFPE 4-µm thin tissue sections were used to perform immunohistochemistry in a Leica Bond Max
automated stainer (Leica Biosystems). The tissue sections were deparaffinized and rehydrated following
the Leica Bond protocol. Antigen retrieval was performed with EDTA buffer (MXB Biotechoiogles, pH 9.0,
MVS-0098), or citrate sodium buffer (MXB Biotechoiogles, pH 6.0, MVS-0100) for 20 min. Slides were
washed, blocked, and then the primary antibody was incubated overnight at 4℃ temperature. The
primary antibody was detected using the DAB Detection Kit (MXB Biotechoiogles, Kit-0014). The slides
were counterstained with haematoxylin, dehydrated and capped with coverslip.
Western blot assay
Page 5/23
The proteins were extracted from the breast cancer specimens and cells using RIPA lysis buffer
(Beyotime, Shanghai, China). BCA Protein Assay Kit was used to evaluate the protein concentration. The
protein was separated from the sample buffer using SDS-PAGE, transferred into PVDF membranes and
blocked with 5% skim milk for 1 h. The primary antibodies were anti-LRRC15 (Abcam, ab150376, 1:1000),
anti-GAPDH (Abcam, ab82485, 1:1000). The member was incubated at 4℃ overnight and then incubated
with horseradish peroxidase-conjugated secondary antibodies. Immunoblots were visualized by BIO-RAD
molecular imager.
SiRNA transfection
SiRNA transfection assay was performed by the transfection was performed using Lipofectamine™ 3000
(Thermofisher, L300015) according to the Lipofectamine™ 3000 Reagent protocol. MDA-MB-231 cells
were transfected with siRNA Ctrl or mixture of three LRRC15 siRNAs, the final concentration of siRNA
used was 50 nM. After 48 h of transfection, the cells were harvested and then subjected to the follow-up
experiments. The LRRC15 siRNA sequence was designed and confirmed as 5’GTAGGACTCACGAAGCTCA-3’ (Ribobio, siG000131578A).
Cell viability assay
Cell viability assay was conducted using a CCK-8 (Cell Counting Kit-8) assay kit (X-Bio, Technology).
Briefly, 8x103 cells/well were plated in 96-well plates and 10 µL CCK8 solution was added into each well 2
h before detection at the indicated time points. Absorbance was measured at 450 nm by SpectraMax®
Absorbance Reader.
Cell cycle assessed by flow-cytometric assay
Distribution of cells in cell cycle was analysed by flow cytometry. MDA-MB-231 cells were seeded in a 6well cell culture plate and treated with different concentration of CoCl2 to induce hypoxia (10, 20, 25 and
50 µM for MDA-MB-231 cells). The cell harvested followed by fixation in 70% chilled ethanol. Staining of
the cells was carried out with PI-RNase solution (1 mg/mL PI, 0.1% V/V Triton X-100 and 10 mg/mL
RNase). The reading was done on a Beckman Coulter Gallios flow cytometer. A Beckman Coulter Gallios
flow cytometer was used for data analysis.
Statistical analysis
In this study, Student’s t-test was used to generate a P-value and the P-value cutoff was 0.05. In oncomine
database, a p value of 0.05 and a fold change of 2 were set as the significance thresholds. Axis units
from TCGA data mining were Log2 (TPM + 1). Spearman correlation analysis was adopted to evaluate
the correlation of LRRC15 expression and immune markers. P-values <0.05 were considered as
statistically significant.
Results
LRRC15 is Highly Upregulated in Breast cancer
Page 6/23
We firstly assessed the expression of LRRC15 in diverse malignancies and normal tissue types using the
Oncomine database, revealing that expression of this gene was elevated relative to normal tissue controls
for breast particularly (Figure 1A). Using RNA expression profiling from TCGA database, we identified
LRRC15 as highly expressed in multiple solid tumor indications together with limited normal tissue
expression. Notably, particularly elevated abundance of LRRC15 mRNA was observed in BCa (Figure 1B).
Therefore, that's why we want to study LRRC15 in BCa. Furthermore, based on data mining, we found that
LRRC15 has higher expression in cancerous tissues than in normal tissues both in mRNA and protein
expression level (Figure 1C-D). Consistently, it is also proven in the immunohistochemical results of BCa
patients from HPA database (Figure 1G). We also analyzed LRRC15 protein levels in BCa cell lines.
Through the further experiment verification, we found LRRC15 was indeed highly in BCa cell lines
compared to MCF-10A cell in protein expression level. We performed Western blot analysis to examine the
LRRC15 expression in clinical tissues, 7 pairs of tumor and paracancerous tissues were observed. As
shown in Figure 1H, LRRC15 protein level was significantly elevated in tumor tissues compared with
matched paracancerous tissues. Collectively, we concluded that LRRC15 expression was significantly
elevated in BCa.
Subtype Analysis Of Lrrc15 In Bca
Further, when sorting the patients by subgroups, all subunits of the LRRC15 were still significantly upregulated in different molecular subtypes compared with paracancerous samples. As shown in Figure 2A,
the highest expression levels of LRRC15 were observed in luminal and Her-2 positive subtype samples
compared with TNBC (Luminal vs TNBC P=1.62e-12; HER2 Positive vs TNBC 3.96e-03). Furthermore,
LRRC15 still showed obviously strong expression in luminal and Her-2 positive BCa compared with seven
TNBC subtypes, including TNBC-BL1 (TNBC Basal-like 1), TNBC-BL2 (TNBC Basal-like 2), TNBC-IM (TNBC
Immunomodulatory), TNBC-M (TNBC Mesenchymal), TNBC-MSL (TNBC mesenchymal stem-like), TNBCLAR (TNBC luminal androgen receptor) and TNBC-UNS (TNBC unspecified) (Figure 2B). IHC assays were
performed to observe differences in LRRC15 expression levels among different molecular subtypes of
BCa, and the highest protein expressions of LRRC15 were observed in luminal B subtypes (Figure 2C-G).
In addition, we also observed the expression of LRRC15 in different pathological types from oncomine
database, which was highly expressed in both invasive ductal carcinoma and invasive lobular carcinoma
(Figure 2H-K).
Association of mRNA expression of LRRC15 with prognosis and clinicopathological parameters in BCa
patients
We used Kaplan-Meier plotter and HPA database to analyze the prognostic values of the mRNA
expression of LRRC15 in BCa patients. Higher mRNA expression of LRRC15 was related to poorer OS in
BCa patients (HR=1.56, 95% CI: 1.01-2.42, and P=0.045). We also HPA database analyze the prognostic
values of the protein expression of LRRC15 in BCa patients. Overexpression of LRRC15 was significantly
associated with unfavorable prognosis in BCa patients (P=0.032). Further, we investigated molecular
Page 7/23
characterization of prognostic interactions. As were shown in Fig. 3D-F, higher mRNA expression of
LRRC15 was significantly associated with shorter OS of BCa patients in luminal B (HR=1.57, 95% CI: 1.022.41, and P=0.0392), Her-2 (HR=1.52, 95% CI: 1.13-2.05, and P=0.0052) and TNBC (HR=14.07, 95% CI:
1.85->100, and P=0.0107), while LRRC15 mRNA expression showed no correlation with prognosis of
luminal A BCa patients (Fig. 3C). Furthermore, we evaluate the association between LRRC15 expression
and clinicopathological parameters in BCa patients, the results showed that distant metastasis was
associated with high expression of LRRC15 (Fig. 3G).
Correlation Analysis Between Lrrc15 Expression And
Immunity In Bca
We primarily aimed to investigate the relationship between LRRC15 expression and immunity in TME of
BCa. We analyzed the correlation between LRRC15 expression and 6 types of infiltrating immune cells
from TIMER database (B cells, CD4+ T cells, CD8+ T cells, neutrophils, macrophages, and dendritic cells).
The results showed that LRRC15 expression levels had a significantly negative correlation with tumor
purity (r = -0.206, P = 5.05e-11)and a significantly positive correlation with infiltrating levels of B cells (r =
0.101, P = 1.58e-03), CD8+ T cells (r = 0.248, P = 4.28e-15), CD4+ T cells (r = 0.2, P = 3.55e-10),
macrophages (r = 0.436, P = 7.60e-47), neutrophils (r = 0.262, P = 2.17e-16), and dendritic cells (r = 0.287,
P = 1.54e-19) in breast cancer (Figure 4A). Furthermore, we investigated whether LRRC15 expression was
correlated with immune infiltration levels in different BCa subtypes, and the correlations were not all the
same (Figure 4B–4D). Analogously, in luminal BCa, LRRC15 expression levels were negatively correlated
with tumor purity and positively correlated with infiltrating levels of these 6 types of infiltrating immune
cells. In TNBC, LRRC15 expression levels were positively correlated with infiltrating levels of 4 types of
infiltrating immune cells, including CD4+ T cells, macrophages, neutrophils, and dendritic cells.
Divertingly, in HER2+ BCa, LRRC15 expression levels had no significant correlations with CD8+ T cells,
CD4+ T cells, macrophages, neutrophils, and dendritic cells. In addition, copy number of LRRC15
expression was linked with infiltrating levels of CD4+ T cells (Figure 4E). Expression of LRRC15 was
negatively correlated with TMB in BCa, whereas no significant correlation with MSI (Figure 4F-G). LRRC15
expression was positively correlated with CD274, CTLA4, HAVCR2, PDCD1LG2 and SIGLEC15, and
negatively correlated with LAG3. Expression of LRRC15 had significantly positive correlation with
follicular helper T cell and activated NK cell, and had moderately positive correlation with CD8+T cell,
memory activated CD4+ T cell, activated myeloid dendritic cell, resting NK cell, plasma B cell and memory
B cell. Expression of LRRC15 was negatively correlated with CD4+ memory resting T cell, active mast cell
and M2 macrophage. We next further explored the link between LRRC15 expression and sets of
immunological markers in GEPIA. These markers were used to characterize immune cells, including CD8+
T cell, T cell (general), B cell, monocyte, TAM, M2 Macrophage, neutrophils, natural killer cell, dendritic cell,
Th1, Th2, Tfh, Th17, Treg, T cell exhaustion in BCa. LRRC15 was positively correlated with Treg markers
(FOXP3, CCR8, STAT5B, TGFβ) in BCa tissues. Detailed findings in BCa and normal tissues are compiled
in Table 1. Hence, these results suggests that LRRC15 was a vital factor in tumor immune
Page 8/23
microenvironment of BCa, potentially playing a significant role in governing immune cell infiltration and
thus representing a meritorious prognostic biomarker in BCa patients.
Page 9/23
Table 1
Correlation analysis between LRRC15 and genes markers of immune cells in
GEPIA
Description
CD8+ T cell
T cell (general)
B cell
Monocyte
TAM
Gene markers
Tumor
Normal
cor
P
cor
P
CD8A
-0.039
0.19
0.0014
0.99
CD8B
−0.097
**
-0.04
0.67
CD3D
-0.072
*
0.071
0.45
CD3E
0.045
0.14
−0.091
0.34
CD2
-0.022
0.47
0.11
0.23
CD19
-0.077
*
0.13
0.17
CD79A
-0.086
**
0.15
0.11
CD86
0.2
***
0.00048
0.32
CD115
0.24
***
0.29
**
CCL2
0.039
0.2
0.26
**
CD68
0.23
***
0.15
0.13
0.1
***
0.2
*
IRF5
-0.0047
0.88
0.22
*
CD163
0.036
0.24
0.16
0.092
VSIG4
0.15
***
0.19
*
MS4A4A
0.12
***
0.23
*
CD66b
−0.0087
0.78
0.069
0.47
CD11b
0.24
***
0.16
0.09
CCR7
-0.057
0.062
0.15
0.11
KIR2DL1
-0.032
0.29
0.099
0.3
KIR2DL3
-0.068
*
0.23
*
KIR2DL4
-0.014
***
0.13
0.16
KIR3DL1
-0.053
0.083
0.18
0.062
KIR3DL2
-0.069
*
0.29
**
IL10
M2 Macrophage
Neutrophils
Natural killer cell
Page 10/23
Dendritic cell
Th1
Th2
Tfh
Th17
Treg
T cell exhaustion
KIR3DL3
0.041
*
-0.025
0.79
KIR2DS4
-0.071
*
0.06
0.53
HLA-DPB1
0.14
***
0.06
0.53
HLA-DQB1
0.043
0.15
0.15
0.1
HLA-DPA1
0.19
***
0.26
**
BDCA-1
0.013
0.68
0.28
**
BDCA-4
0.44
0
0.16
0.084
CD11c
0.19
***
0.1
0.28
T-bet
-0.071
*
0.23
*
STAT4
0.0082
0.79
0.13
0.17
STAT1
0.0074
0.81
0.041
0.67
IFN-γ
-0.088
**
0.18
0.053
TNF-α
-0.046
0.13
0.16
0.096
GATA3
0.023
0.44
-0.013
0.89
STAT6
0.14
***
0.28
**
STAT5A
0.075
*
-0.14
0.13
IL13
-0.0012
0.97
0.24
**
BCL6
0.092
**
0.0012
0.99
IL21
-0.049
0.1
0.12
0.21
STAT3
0.22
***
0.22
*
IL17A
-0.042
0.17
0.15
0.12
FOXP3
0.067
*
0.051
0.6
CCR8
0.12
**
0.16
0.083
STAT5B
0.14
***
0.055
0.57
TGFβ
0.39
***
0.3
**
PD-1
-0.012
0.69
0.27
**
CTLA4
-0.051
0.095
0.21
0.029
LAG3
-0.13
***
0.23
*
TIM-3
0.33
***
0.18
0.056
Page 11/23
GZMB
-0.17
***
0.25
**
The cancer-promoting role of LRRC15 in tumor microenvironment of BCa
The above results strongly showed that LRRC15 might have a tumor-promoting effect in BCa. We then
focused on its potential oncogenic roles by conducting siRNA-mediated loss-of-function approach. We
selected MDA-MB-231 and BT-474 cells for knockdown experiments for the relatively higher endogenous
LRRC15 in these cell lines (Figure 1D). Co-expression And Pathway Enrichment Analyses Of Lrrc15
A gene-gene interaction network for LRRC15 was constructed, and their functions were analyzed using
the GeneMANIA database (Fig. 6A). The 12 central nodes representing LRRC15 were surrounded by 20
nodes representing genes that greatly correlated with the family in terms of predictions, physical
interactions, co-expression, genetic interactions, and co-localization. The top five genes displaying the
greatest correlations with the LRRC15 included TARDBP, ANXA5, KHDRBS2, USP15 and DDIT4L. LRRC15
was correlated with these genes mainly in terms of physical interactions. Further GSEA analysis revealed
that high expression of LRRC15 showed the greatest correlation with ECM receptor interaction, focal
adhesion, regulation of actin cytoskeleton, and TGF Beta signaling pathway. Besides, low expression of
LRRC15 showed the greatest correlation with DNA replication, homologous recombination, oxidative
phosphorylation and ribosome. Additionally, we further performed transcriptome RNA sequencing to
identify the DEGs (differentially expressed genes) between si-LRRC15 and si-NC MDA-MB-231 cells. GO
(gene ontology) analysis results showed DEGs were enriched in extracellular space, protein
heterodimerization and immune response. In addition, KEGG (Kyoto Encyclopedia of Gene and Genome)
analysis showed DEGs were enriched in alcoholism, systemic lupus erythematosus and transcriptional
misregulation in cancer.
Discussion
Page 12/23
Herein, we conducted a comprehensive assessment of the correlation between LRRC15 and TME of BCa.
LRRC15 primarily had effects on regulation tumor growth and immunity of BCa. ABBV-085, a novel
antibody-drug conjugate (ADC) targeting LRRC15, is conjugated to the tubulin inhibitor monomethyl
auristatin E (MMAE) and has indicated distinct anti-tumor activity in multiple tumors [22–24]. LRRC15
played a vital role in regulating immune cell infiltration in BCa, and the degree of immune infiltration
remarkably influenced cancer survival. Previous studies demonstrated that immune cells in the TME
could be used in the prognostic assessment of multiple malignancies, such as BCa, glioblastoma, and
melanoma [25–27]. Tang et.al reported that LRRC15 acted in collusion with macrophages in the TME of
glioblastoma to prompt poor prognosis, especially in mesenchymal subtype [28]. This was consistent
with our analysis results: macrophages were most associated with LRRC15 among various immune
infiltration cells. It indicated targeting LRRC15 to enhance macrophages-based immunosuppressive
effects might be promising strategies for cancer treatments. Furthermore, other MMAE-linked ADCs have
been assumed to augment immunogenic cell death and have displayed evidence of enhancive response
rates when combined with immunomodulatory anti-PD-1 antibody therapy [29]. Therefore, a promising
hypothesis for future research is to combine ABBV-085 with anti-PD-1 treatment to augment tumor
responses.
LRRC15 is capable of binding collagen, which sustains its role in tissue structure and cell-to-cell adhesion
[7]. It has been proposed that extracellular matrix (ECM) proteins generated by CAF prevent the effective
uptake of traditional chemotherapeutics, and contribute to the immunosuppressive environment and
chemoresistance in most solid tumors [8, 30, 31]. As a biomarker of CAF, LRRC15 might be an important
contributor, and also an effective therapeutic target of immunosuppression and chemoresistance.
LRRC15 was also an essential mediator for osteogenesis of mesenchymal stem cells (MSCs) through
repressing the NF-κB pathway in a p65-dependent manner, giving a novel hint for MSC-mediated bone
regeneration [32].
Conclusions
In summary, our results provided insights into understanding the potential role of LRRC15 mRNA in tumor
immunology, prognostic value and enrichment pathways. LRRC15 mRNA level was correlated with
immune infiltrating levels, indicating that it might participate in tumor progression and exert
immunotherapeutic effects on BCa. The potential for LRRC15 inhibitors to interfere with tumor immune
microenviroment deserved further and deepgoing investigations.
Declarations
Author Contributions
Concept, drafting: LL and XWX. Figure making: FYJ, LMX, and WGQ. Figure making: SXY. Revision CW
and LJY. Supervision: TJH. All authors contributed to the article and approved the submitted version.
Page 13/23
Acknowledgements
None.
Competing interests
The authors declare no potential conflict of interest. All the datasets were retrieved from the published
literature, so it was confirmed that all written informed consent was obtained.
Funding statement
This research was supported by the National Key Research and Development Program of China (No.
2016YFC0905900), National Natural Science Foundation of China (No. 81872365, No. 81902987) and
Jiangsu Provincial Key Research Development Program (No. BE2019731).
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NY) 2015, 347(6220):1260419.
20. Li T, Fan J, Wang B, Traugh N, Chen Q, Liu JS, Li B, Liu XS: TIMER: A Web Server for Comprehensive
Analysis of Tumor-Infiltrating Immune Cells. Cancer research 2017, 77(21):e108-e110.
Page 15/23
21. Warde-Farley D, Donaldson SL, Comes O, Zuberi K, Badrawi R, Chao P, Franz M, Grouios C, Kazi F,
Lopes CT et al: The GeneMANIA prediction server: biological network integration for gene prioritization
and predicting gene function. Nucleic Acids Res 2010, 38(Web Server issue):W214-220.
22. Ben-Ami E, Perret R, Huang Y, Courgeon F, Gokhale PC, Laroche-Clary A, Eschle BK, Velasco V, Le
Loarer F, Algeo MP et al: LRRC15 Targeting in Soft-Tissue Sarcomas: Biological and Clinical Implications.
Cancers 2020, 12(3).
23. Slemmons KK, Mukherjee S, Meltzer P, Purcell JW, Helman LJ: LRRC15 antibody-drug conjugates
show promise as osteosarcoma therapeutics in preclinical studies. Pediatric blood & cancer 2021,
68(2):e28771.
24. Hingorani P, Roth ME, Wang Y, Zhang W, Gill JB, Harrison DJ, Teicher B, Erickson S, Gatto G, Smith
MA et al: ABBV-085, Antibody-Drug Conjugate Targeting LRRC15, Is Effective in Osteosarcoma: A Report
by the Pediatric Preclinical Testing Consortium. Molecular cancer therapeutics 2021, 20(3):535-540.
25. Fortis SP, Mahaira LG, Anastasopoulou EA, Voutsas IF, Perez SA, Baxevanis CN: Immune profiling of
melanoma tumors reflecting aggressiveness in a preclinical model. Cancer immunology, immunotherapy :
CII 2017, 66(12):1631-1642.
26. Priedigkeit N, Watters RJ, Lucas PC, Basudan A, Bhargava R, Horne W, Kolls JK, Fang Z, Rosenzweig
MQ, Brufsky AM et al: Exome-capture RNA sequencing of decade-old breast cancers and matched
decalcified bone metastases. JCI Insight 2017, 2(17).
27. Jia D, Li S, Li D, Xue H, Yang D, Liu Y: Mining TCGA database for genes of prognostic value in
glioblastoma microenvironment. Aging 2018, 10(4):592-605.
28. Tang H, Liu W, Xu Z, Zhao J, Wang W, Yu Z, Wei M: Integrated microenvironment-associated
genomic profiles identify LRRC15 mediating recurrent glioblastoma-associated macrophages infiltration.
Journal of cellular and molecular medicine 2021, 25(12):5534-5546.
29. Herrera AF, Moskowitz AJ, Bartlett NL, Vose JM, Ramchandren R, Feldman TA, LaCasce AS, Ansell
SM, Moskowitz CH, Fenton K et al: Interim results of brentuximab vedotin in combination with nivolumab
in patients with relapsed or refractory Hodgkin lymphoma. Blood 2018, 131(11):1183-1194.
30. Turley SJ, Cremasco V, Astarita JL: Immunological hallmarks of stromal cells in the tumour
microenvironment. Nature reviews Immunology 2015, 15(11):669-682.
31. Olive KP, Jacobetz MA, Davidson CJ, Gopinathan A, McIntyre D, Honess D, Madhu B, Goldgraben MA,
Caldwell ME, Allard D et al: Inhibition of Hedgehog signaling enhances delivery of chemotherapy in a
mouse model of pancreatic cancer. Science (New York, NY) 2009, 324(5933):1457-1461.
32. Wang Y, Liu Y, Zhang M, Lv L, Zhang X, Zhang P, Zhou Y: LRRC15 promotes osteogenic
differentiation of mesenchymal stem cells by modulating p65 cytoplasmic/nuclear translocation. Stem
Page 16/23
cell research & therapy 2018, 9(1):65.
Figures
Figure 1
LRRC15 was overexpressed in BCa cells and tissues (A) LRRC15 mRNA expression level in data sets of
diverse cancers compared with normal tissues from the Oncomine database. The numbers in the colored
Page 17/23
squares display the number of the involved studies. The color intensity is directly proportional to the high
or low expression level, respectively. Red indicates high expression, while blue indicates low expression.
(B) Relative mRNA expression of LRRC15 in pan-cancer based on TCGA data mining. (C) mRNA
expression of LRRC15 in from GEPIA database. (D) Protein expression of LRRC15 in BCa from UALCAN
database. (E-F) Endogenous expression of LRRC15 in MCF-10A and multiple BCa cell lines were detected
by Western blotting. (G) Representative immunohistochemistry images of LRRC15 in BCa and normal
tissues derived from the HPA database. (H) LRRC15 protein levels in 7 paired BCa tissues and their
adjacent noncancerous tissues were examined by Western blotting.
Page 18/23
Figure 2
Expression levels of LRRC15 in BCa tissues was from different subtypes (A) LRRC15 mRNA expression in
four major molecular subtypes of BCa from UALCAN database. (B) LRRC15 mRNA expression in four
major molecular subtypes of BCa from UALCAN database. Representative images of
immunohistochemical staining in (C) normal tissues (D) luminal A (E) luminal B (F) HER2 (G)TNBC BCa
Page 19/23
tissues. LRRC15 mRNA expression in (H) invasive ductal BCa (I) invasive lobular BCa (J) invasive ductal
and lobular BCa (K) invasive BCa from oncomine database.
Figure 3
Association of mRNA expression of LRRC15 with prognosis and clinicopathological parameters in BCa
patients (A) Relationship between LRRC15 mRNA expression and the OS of BCa patients. (B)
Relationship between LRRC15 protein expression and the OS of BCa patients. Prognosis of LRRC15
mRNA expression in different molecular subtypes of BCa: (C) luminal A (D) luminal B (E) HER2 (F) TNBC
Page 20/23
BCa tissues. (G) Association between LRRC15 expression and clinicopathological parameters in BCa
patients.
Figure 4
Correlation of LRRC15 expression with immunity in BCa. Immune infiltration analysis based on TIMER
database in (A) Breast invasive carcinoma, (B) TNBC, (C) HER2 and (D) luminal subtypes. (E) Effect of
CNA of LRRC15 expression on immune infiltration level in BCa. Correlation of LRRC15 expression with (F)
Page 21/23
MSI (G) TMB, (H) immune checkpoints and (I) 22 immune cell subtypes. TMB, tumor mutational burden;
MSI, microsatellite instability.
Figure 5
LRRC15 was positively correlated with cell proliferation of BCa. Cell viability was examined CCK-8 assay
in (A) MDA-MB-231 cells and (B) BT-474 cells. (C) Representative examples of LRRC15
immunohistochemistry (IHC) staining in BCa tissues. IHC analysis of LRRC15 and Ki-67 levels in BCa
tissues with low or high expression. It revealed that LRRC15 was positively correlated with Ki-67 in situ.
(D-F) Cell cycle analyzed by flow cytometry in BCa.
Page 22/23
Figure 6
Gene-gene interaction network and enrichment pathway of LRRC15 in BCa patients (A) Gene–gene
interaction network of LRRC15 from Genemania database. Each node represents a gene, and the internode connection lines represent the types of gene-gene interactions. The node size represents the
strength of interactions. The node color represents the possible functions of respective genes. (B) The
enriched KEGG pathway sets by the high LRRC15 expression. (C) The enriched KEGG pathway sets by the
low LRRC15 expression. (D) GO enrichment analysis of DEGs between si-LRRC15 and si-NC MDA-MB-231
cells. (E) KEGG enrichment analysis of DEGs between si-LRRC15 and si-NC MDA-MB-231 cells. GO: gene
ontology; KEGG: Kyoto Encyclopedia of Gene and Genome; DEGs: Differentially expressed genes.
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Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 1Formerly, Professor of Mathematics, Federal University of Santa Catarina, Florianópolis, SC, Brazil (1978-2012).
Also worked at Delhi University, India (1976-1978).
E-mail: ijtaneja@gmail.com; Web-sites: http://inderjtaneja.com; http://numbers-magic.com;
Twitter: @IJTANEJA; Instagram: @crazynumbers. Figured Magic Squares of Order 30 Using Bordered
Magic Rectangles: A Systematic Procedure Inder J. Taneja1 1
Introduction 2
Magic Squares of Order 30
4
2.1
Block-Wise and Bordered Square of Order 30 . . . . . . . . . . . . . . . . . . . . . . . 4
2.2
Magic Squares of Order 30 With BMRs . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
2.3
Cornered Magic Squares of Order 4 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
2.4
Closed Border of Order 4 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
2.5
Cornered Magic Squares of Order 6 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
2.6
Closed Border of Order 6 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
2.7
Cornered Magic Squares of Order 8 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96
2.8
Closed Border of Order 8 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
2.9
Equal Sums Blocks of Order 10
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
2.10 Closed Border of Order 10 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
2.11 Cornered Magic Squares of Order 12 . Abstract Recently, author constructed even order magic squares from orders 6 to 20 with different styles
and models, for examples the order 20 is with 1616 magic squares, order 18 with 810 magic
squares, etc. These can be seen at [31, 32, 33, 33, 34, 35, 36, 37]. The aim is to proceed for
the further orders of magic squares. In this work there are few examples of magic squares given
as figures of order 30. A systematic procedure to construct these magic squares is given. It is
based on the magic squares of orders 4, 6, 8 etc. at corners and small blocks of bordered magic
rectangles forming external borders. Then the internal borders are filled with previous known
magic squares. The presentations is in figures instead of numbers. The readers can find replies in
numbers from references given above. For the orders multiples of 4, we can always write magic
squares with equal sums blocks of magic squares of order 4. This procedure is very helpful for the
orders of type 2p, where p is a prime number, for examples, 14, 22, 26, 34, 38, etc. For the orders
like 18, 30, etc. we can make good external blocks with order 4, and for orders like 16, 20, 28, 32,
etc. we can make good external borders of order 6, and so on. 1 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja https://inderjtaneja.com; https://numbers-magic.com; 1
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
2.12 Closed Border of Order 12 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158
2.13 Magic Squares of Orders 14, 16 and BMRs of Order 14 × 16 . . . . . . . . . . . . . . . 162
2.14 Extra Examples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 168
3
Appendix
173
4
Author’s Contribution to Magic Squares and Recreation Numbers
174 1
Introduction magic sums of order n of consecutive numbers from 1 to n2 is given by
Sn×n := n × (1 + n2)
2
, n ≥3. (1) The magic sums of order n of consecutive numbers from 1 to n2 The magic sums of order n of consecutive numbers from 1 to n2 is given by Sn×n := n × (1 + n2)
2
, n ≥3. (1) Recently, the author [31, 32, 33, 34, 35, 36, 37] constructed magic squares of even orders from 8
to 20 using bordered magic rectangles. This construction is based on two aspects: (i) Using magic rectangles or bordered magic rectangles. (ii) Using algebraic formula like (a + b)2, a ̸= b. For the above magic squares no construction procedure is explained. The aim is to proceed
further orders of magic squares. In this work, a systematic procedure to construct these magic
squares is given. It is based on the magic squares and bordered magic rectangles (BMR) of orders 4,
6, 8 etc forming external borders. Then the internal borders are filled with previous known magic
squares. For the orders multiples of 4, we can always write magic squares with equal sums blocks
of magic squares of order 4. This procedure is very helpful for the orders of type 2p, where p is
a prime number, for examples, 14, 22, 26, 34, 38, etc. For the orders like 18, 30, etc.,we can make
good external blocks with order 4, and for orders like 16, 20, 28, 32, etc. we can make good external
borders of order 6, and so on. There is no explainations for the orders 6, 8, 10 and 12. The real
construction starts from the order 14. For the above magic squares no construction procedure
further orders of magic squares. In this work, a systemati
squares is given. It is based on the magic squares and bordere
6, 8 etc forming external borders. Then the internal borders
squares. For the orders multiples of 4, we can always write m
of magic squares of order 4. This procedure is very helpful
a prime number, for examples, 14, 22, 26, 34, 38, etc. For th
good external blocks with order 4, and for orders like 16, 20, 2
borders of order 6, and so on. There is no explainations for
construction starts from the order 14. 2
Magic Squares of Order 30 2 2 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
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Introduction The whole the work is done manually, without use of any programming language, except for
the constructions of small blocks of bordered magic rectangles. This construction is based on
the software due to H. While. Later, these BRMs are readopted according to distribution of each
magic square. The distribution of magic squares or bordered magic rectangles is based on half-
sequential numbers. By half-sequential numbers we understand that the total numbers in each
case are divided in two equal parts. The first part is one sequence and the second part is another
sequence. Due to half-sequential numbers, it is not possible to construct all orders bordered magic
rectangles. In Appendix 3, there are tables showing the existence of these bordered magic rectan-
gles for half-sequential. For simplicity, we shall write BMR as bordered magic rectangle. This
work is for order 30. The previous works can be seen at [38, 39, 40, 41, 42, ?]. 3 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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remove external borders in each case, still we are left with bordered magic squares and bordered magic
rectangles respectively with sequential number entries. The whole work is in figures instead of numbers. In the same magic square the small equal figures represents equal sums magic squares and/or equal sums
bordered magic rectangles. The whole work is done manually Remark 1. The bordered magic squares and bordered magic rectangles are with the property that if we
remove external borders in each case, still we are left with bordered magic squares and bordered magic
rectangles respectively with sequential number entries. The whole work is in figures instead of numbers. In the same magic square the small equal figures represents equal sums magic squares and/or equal sums
bordered magic rectangles. The whole work is done manually 2
Magic Squares of Order 30 This section brings in figures (without numbers) magic squares of order 30. In some cases, the idea
of constructions are explained. It is based on cornered magic squares. 2.1
Block-Wise and Bordered Square of Order 30 Below are magic squares of order 30 already known in the literature. For more details refer author’s
work [22, 24]. 4 4 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
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https://inderjtaneja.com; https://numbers-magic.com; Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Magic Squares of Order 30 With BMRs Below are examples of magic squares of order 28 made with BMRs. The construction in each case
is individual. 12 12 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
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Cornered Magic Squares of Order 4 Let’s consider an external border, where there are 4 magic squares of order 4 at the corners. Let’s
make an external border by putting BMR of order 4 × 22 in each row and column. In the middle
we are left with block of order 22. Writing this middle blocks with different types of magic squares
of order 22, we get magic squares of order 30. See below few examples: 17 17 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 2.3
Cornered Magic Squares of Order 4 1-179, https://doi.org/10.5281/zenodo.7390705 24 24 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 25 25 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 26 26 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 27 27 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 28 28 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja J
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 29 29 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://n tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 30 30 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https://numbers-m Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 31 31 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https://numbers-m Inder J. Taneja
h
//
d J
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https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 2.3
Cornered Magic Squares of Order 4 1-179, https://doi.org/10.5281/zenodo.7390705 32 32 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
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https://inderjtaneja.com; https://numbers-magic.com; tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 33 33 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https://number Inder J. Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 34 34 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 35 35 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja j
tps://inderjtaneja.com; https://numbers-magic.com;
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 36 36 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 37 37 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 38 38 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 39 39 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https://numbers- Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 40 40 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https://numbers-m Inder J. 2.3
Cornered Magic Squares of Order 4 Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 41 41 41 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 42 42 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 43 43 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja 2.4
Closed Border of Order 4 The external border is made with 4 BMRs of order 4 × 26. We are left with inner block of order 22. Writing this middle blocks with different types of magic squares of order 22, we get magic squares
of order 30. See below few examples: 44 44 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https://numbers- Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 45 45 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja Inder J. Taneja
h
//
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https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 46 46 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.c Inder J. Taneja
h
//i d j Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 47 47 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com Inder J. Taneja
h
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https://inderjtaneja.com; https://numbers-magic.com; tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 48 48 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.co Inder J. Taneja
h
//i d j J
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https://inderjtaneja.com; https://numbers-magic.com; tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 49 49 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.c Inder J. Taneja
h
//i d j Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 50 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https:// Inder J. 2.4
Closed Border of Order 4 Taneja
h
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 51 51 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.c Inder J. Taneja
h
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https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 52 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja J
j
tps://inderjtaneja.com; https://numbers-magic.com;
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q
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 53 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja Inder J. Taneja
h
//i d j tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 54 54 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; h Inder J. Taneja
h
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d J
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https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 55 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtane Inder J. Taneja
h
//i d j ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 56 56 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https Inder J. Taneja
h
//i d j J
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https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 57 57 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.co Inder J. Taneja
h
//i d j J
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https://inderjtaneja.com; https://numbers-magic.com; tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 58 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; http Inder J. 2.4
Closed Border of Order 4 Taneja
h
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https://inderjtaneja.com; https://numbers-magic.com; tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 59 59 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.c Inder J. Taneja
h
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d J
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https://inderjtaneja.com; https://numbers-magic.com; tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 60 60 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
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d Inder J. Taneja
https://inderjtaneja.com J
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https://inderjtaneja.com; https://numbers-magic.com; ttps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 61 61 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; Inder J. Taneja
h
//i d j Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 62 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja J
j
ttps://inderjtaneja.com; https://numbers-magic.com;
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 63 63 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja J
j
ttps://inderjtaneja.com; https://numbers-magic.com;
g
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q
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 64 64 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja 2.5
Cornered Magic Squares of Order 6 In view of Fig. 4 given in subsection 2.1, we don’t require to make cornered blocks of order 6. But
we have considered below just as an extra option. Let’s consider an external border, where there
are 8 magic squares of order 6 and 4 BMRs of order 6 × 12. This gives us an external border with
cornered magic squares of order 6. In the middle we are left with block of order 18. Writing this
middle blocks with different types of magic squares of order 18, we get magic squares of order 30. See below few examples: 65 65 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 66 66 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 67 67 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https: Inder J. Taneja
h
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d Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 68 68 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
//
d Inder J. Taneja
https://inderjtaneja.com; h j
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1 179, https://doi.org/10.5281/zenodo.7390705 69 69 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtane Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; j
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1 179, https://doi.org/10.5281/zenodo.7390705 70 70 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 71 71 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 2.5
Cornered Magic Squares of Order 6 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 72 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 73 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; h ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 74 74 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; j
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1 179, https://doi.org/10.5281/zenodo.7390705 75 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtane Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1 179, https://doi.org/10.5281/zenodo.7390705 76 76 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; h Inder J. Taneja
h
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d ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 77 77 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; h Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 78 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 79 79 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 80 80 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. 2.5
Cornered Magic Squares of Order 6 Taneja
https://inderjtanej Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 81 81 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtanej Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 82 82 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; j
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 83 83 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja 2.6
Closed Border of Order 6 Let’s write a closed external border formed by 4 blocks of equal sums BMRs of order 6 × 24. In the
middle we are left with blocks of order 18. Writing this middle blocks with different types of magic
squares of order 18, we get magic squares of order 30. See below few examples: 84 84 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
//
d ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 85 85 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtanej Inder J. Taneja
h
//i d j Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 https://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 86 86 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.co Inder J. Taneja
h
//i d j s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 https://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 87 87 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com Inder J. Taneja
h
//i d j ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 88 88 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
//i d j Inder J. Taneja
https://inderjtaneja. ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 89 89 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; ht Inder J. Taneja
h
//i d j ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 90 90 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com Inder J. 2.6
Closed Border of Order 6 Taneja
h
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 91 91 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja j
ps://inderjtaneja.com; https://numbers-magic.com;
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 92 92 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
//i d j Inder J. Taneja
https://inderjtaneja.co tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 93 93 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
//i d j Inder J. Taneja
https://inderjtane Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 94 94 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; http Inder J. Taneja
h
//i d j ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 95 95 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja 2.7
Cornered Magic Squares of Order 8 Let’s consider an external border, where there are 4 magic squares of order 8 at the corners. Let’s
make an external border by putting BMRs of order 6 × 14 in each row and column. In the middle
we are left with blocks of order 14. Writing this middle blocks with different types of magic squares
of order 14, we get magic squares of order 30. See below few examples: 96 96 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; https://inderjtaneja.com; https://numbers-magic.com; tps://inderjtaneja.com; https://numbers magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1 179, https://doi.org/10.5281/zenodo.7390705 97 97 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 98 98 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; tps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 99 99 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 100 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; http Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 101 101 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtane Inder J. Taneja
https://inderjtaneja.com; https://n Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 102 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; http Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 2.7
Cornered Magic Squares of Order 8 1-179, https://doi.org/10.5281/zenodo.7390705 103 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 104 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 105 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://n Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 106 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtanej Inder J. Taneja
h
//
d Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1 179, https://doi.org/10.5281/zenodo.7390705 107 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtan Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 108 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
//i d j Inder J. Taneja
https://inderjtaneja s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 109 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtane Inder J. Taneja
h
//i d j https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 110 110 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtane Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; http Inder J. Taneja
h
//
d ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 111 111 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. 2.7
Cornered Magic Squares of Order 8 Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtanej Inder J. Taneja
h
//
d s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 112 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja 2.8
Closed Border of Order 8 Let’s consider an external border, where there are 4 BMRs of order 8 × 22. Putting them in rows
and columns, we get a closed border of order 8. In the middle we are left with blocks of order 14. Writing this middle blocks with different types of magic squares of order 14, we get magic squares
of order 30. See below few examples: 113 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
//
d Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 114 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 115 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
//
d s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 116 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 117 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 118 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja 2.9
Equal Sums Blocks of Order 10 Let’s consider 9 equal sums magic squares of order 10. This gives us magic squares of order 30. See
below few examples: 119 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 120 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 121 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 122 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://numbe Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 123 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtane Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 124 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 125 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 126 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 127 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. 2.9
Equal Sums Blocks of Order 10 Taneja ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 128 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 129 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705
er 10 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja 2.10
Closed Border of Order 10 Let’s consider 4 equal sums BMRs of orders 10 × 20. Putting them in rows and columns, we get a
closed border of order 10. In the middle we are left with blocks of order 10. Writing this middle
blocks with different types of magic squares of order 10, we get magic squares of order 30. See
below few examples: 130 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 131 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 132 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja 2.11
Cornered Magic Squares of Order 12 Let’s consider an external border, where there are 4 magic squares of order 12 at the corners. Let’s
make an external border by putting 2 BMRs of order 4 × 6 in each row and column. In the middle
we are left with blocks of order 4. Writing magic squares of order 12 in different ways, we get magic
squares of order 30. See below few examples: 133 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 135 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1 179, https://doi.org/10.5281/zenodo.7390705 136 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 137 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtane Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 138 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja /inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 139 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 140 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. 2.11
Cornered Magic Squares of Order 12 Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 141 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja s://inderjtaneja.com; https://numbers magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1 179, https://doi.org/10.5281/zenodo.7390705 142 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; //inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 143 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; //inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 144 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 145 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1 179, https://doi.org/10.5281/zenodo.7390705 146 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; //inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 147 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 148 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 149 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. 2.11
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https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 151 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 152 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 153 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1 179, https://doi.org/10.5281/zenodo.7390705 154 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 155 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 156 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https://numbe Inder J. Taneja s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 157 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja 2.12
Closed Border of Order 12 Let’s consider an external border with 8 BMRs of order 6 × 16. In the middle we are left with blocks
of order 4. It gives us a magic square of order 30. See below a single example: 158 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtan Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 159 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.c Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 160 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtaneja.c Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 161 161 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja 2.13
Magic Squares of Orders 14, 16 and BMRs of Order 14 × 16 2.13
Magic Squares of Orders 14, 16 and BMRs of Order 14 × 16 2.13
Magic Squares of Orders 14, 16 and BMRs of Order 14 × 16 Let’s consider different combinations of magic squares of order 14 and 16 with 2 BMRs of order
14 × 16, we get magic squares of order 30. See below few examples: 162 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtan Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; https://inderjtaneja.com; https://numbers-magic.com; ps://inderjtaneja.com; https://numbers magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1 179, https://doi.org/10.5281/zenodo.7390705 163 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
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Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 164 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 165 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
//
d s://inderjtaneja.com; https://numbers magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1 179, https://doi.org/10.5281/zenodo.7390705 166 166 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtaneja.com; https://numbers- Inder J. Taneja
h
//
d J
j
https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 167 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; w are few extra examples of magic squares of order 30 done individually with BMRs. Below are few extra examples of magic squares of order 30 done individually with BMRs. 168 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
h
//
d Inder J. Taneja
https://inderjtanej Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 169 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtane Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 170 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja //inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 171 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja Inder J. Taneja
https://inderjtane Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; https://inderjtaneja.com; https://numbers-magic.com; s://inderjtaneja.com; https://numbers-magic.com;
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 172 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; For author’s contribution to magic squares and recreation num • Inder J. Taneja, Recreation of Numbers, https://inderjtaneja.com/2019/06/27/publications-re
of-numbers/ 3
Appendix Below are tables giving the existence of bordered magic rectangles for half-sequential numbers. Below are tables giving the existence of bordered magic rectangles for half-sequential numbers. 173 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com;
Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705
Order of Magic Square
Bordered Magic Rectangles
36
6 × 36, 10 × 36, 14 × 36, 18 × 36,
22 × 36, 26 × 36, 30 × 36, 34 × 36
38
4 × 38, 8 × 38, 12 × 38, 16 × 38, 20 × 38,
24 × 38, 28 × 38, 32 × 38, 36 × 38
40
6 × 40, 10 × 40, 14 × 40, 18 × 40, 22 × 40,
26 × 40, 30 × 40, 34 × 40, 38 × 40
42
4 × 42, 8 × 42, 12 × 42, 16 × 42, 20 × 42,
24 × 42, 28 × 42, 32 × 42, 36 × 42, 40 × 42
44
6 × 44, 10 × 44, 14 × 44, 18 × 44, 22 × 44, 26 × 44,
30 × 44, 34 × 44, 38 × 44,42 × 44
46
4 × 46, 8 × 46, 12 × 46, 16 × 46, 20 × 46, 24 × 46,
28 × 46, 32 × 46, 36 × 46, 40 × 46, 44 × 46
48
6 × 48, 10 × 48, 14 × 48, 18 × 48, 22 × 48, 26 × 48,
30 × 48, 34 × 48, 38 × 48, 42 × 48, 46 × 48
50
4 × 50, 8 × 50, 12 × 50, 16 × 50, 20 × 50, 24 × 50,
28 × 50, 32 × 50, 36 × 50, 40 × 50, 44 × 50, 48 × 50
4
Author’s Contribution to Magic Squares and Recreation Num- Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
//
d J
j
https://inderjtaneja.com; https://numbers-magic.com;
g
g
q
g
g
g
y
Procedure, Zenodo, December 02, 2022, pp. 3
Appendix 1-179, https://doi.org/10.5281/zenodo.7390705
Order of Magic Square
Bordered Magic Rectangles
36
6 × 36, 10 × 36, 14 × 36, 18 × 36,
22 × 36, 26 × 36, 30 × 36, 34 × 36
38
4 × 38, 8 × 38, 12 × 38, 16 × 38, 20 × 38,
24 × 38, 28 × 38, 32 × 38, 36 × 38
40
6 × 40, 10 × 40, 14 × 40, 18 × 40, 22 × 40,
26 × 40, 30 × 40, 34 × 40, 38 × 40
42
4 × 42, 8 × 42, 12 × 42, 16 × 42, 20 × 42,
24 × 42, 28 × 42, 32 × 42, 36 × 42, 40 × 42
44
6 × 44, 10 × 44, 14 × 44, 18 × 44, 22 × 44, 26 × 44,
30 × 44, 34 × 44, 38 × 44,42 × 44
46
4 × 46, 8 × 46, 12 × 46, 16 × 46, 20 × 46, 24 × 46,
28 × 46, 32 × 46, 36 × 46, 40 × 46, 44 × 46
48
6 × 48, 10 × 48, 14 × 48, 18 × 48, 22 × 48, 26 × 48,
30 × 48, 34 × 48, 38 × 48, 42 × 48, 46 × 48
50
4 × 50, 8 × 50, 12 × 50, 16 × 50, 20 × 50, 24 × 50,
28 × 50, 32 × 50, 36 × 50, 40 × 50, 44 × 50, 48 × 50
4
Author’s Contribution to Magic Squares and Recreation Num-
bers 4
Author’s Contribution to Magic Squares and Recreation Num-
bers For author’s contribution to magic squares and recreation numbers please see the links below: Acknowledgement The bordered magic rectangles are constructed based on the software produced by H. White [1]. The author is thankful to H. White for their valuable help. 174 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
h
//i d j • Bordered Magic Squares [11] Inder J. Taneja, Nested Magic Squares With Perfect Square Sums, Pythagorean Triples, and
Borders Differences, Zenodo, June 14, 2019, pp. 1-59, http://doi.org/10.5281/zenodo.3246586. [12] Inder J. Taneja, Symmetric Properties of Nested Magic Squares, Zenodo, June 29, 2019, pp. 1-55, http://doi.org/10.5281/zenodo.3262170. [13] Inder J. Taneja, General Sum Symmetric and Positive Entries Nested Magic Squares, Zenodo,
July 04, 2019, pp. 1-55, http://doi.org/10.5281/zenodo.3268877. [14] Inder J. Taneja, Bordered Magic Squares With Order Square Magic Sums, Zenodo, January 20,
2020, pp. 1-26, http://doi.org/10.5281/zenodo.3613690. [15] Inder J. Taneja, Fractional and Decimal Type Bordered Magic Squares With Magic Sum 2020. Zenodo, January 20, 2020, pp.1-25. http://doi.org/10.5281/zenodo.3613698. [16] Inder J. Taneja, Fractional and Decimal Type Bordered Magic Squares With Magic Sum 2021,
Zenodo, December 16, 2020, pp. 1-33, http://doi.org/10.5281/zenodo.4327333. [17] Inder J. Taneja, Inder J. Taneja, Block-Wise and Block-Bordered Magic Squares With Magic
Sum 2022, Zenodo, December 28, 2021, pp. 1-38, https://doi.org/10.5281/zenodo.5807789 • Block-Wise Magic Squares [4] Inder J. Taneja, Block-Wise Constructions of Magic and Bimagic Squares of Orders 8 to 108,
May 15, 2019, pp. 1-43, Zenodo, http://doi.org/10.5281/zenodo.2843326. [5] Inder J. Taneja, Block-Wise Equal Sums Pandiagonal Magic Squares of Order 4k, Zenodo,
January 31, 2019, pp. 1-17, http://doi.org/10.5281/zenodo.2554288. [6] Inder J. Taneja, Magic Rectangles in Construction of Block-Wise Pandiagonal Magic Squares,
Zenodo, January 31, 2019, pp. 1-49, http://doi.org/10.5281/zenodo.2554520. [7] Inder J. Taneja, Block-Wise Equal Sums Magic Squares of Orders 3k and 6k, Zenodo, February
1, 2019, pp. 1-55, http://doi.org/10.5281/zenodo.2554895. [8] Inder J. Taneja, Block-Wise Unequal Sums Magic Squares, Zenodo, February 1, 2019, pp. 1-52,
http://doi.org/10.5281/zenodo.2555260. [9] Inder J. Taneja, Block-Wise Magic and Bimagic Squares of Orders 12 to 36, Zenodo, February
1, 2019, pp. 1-53, http://doi.org/10.5281/zenodo.2555343. [10] Inder J. Taneja, Block-Wise Magic and Bimagic Squares of Orders 39 to 45, Zenodo, February
2, 2019, pp. 1-73, http://doi.org/10.5281/zenodo.2555889. 175 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtane • Block-Wise and Block-Bordered Magic Squares • Block-Wise and Block-Bordered Magic Squares [21] Inder J. Taneja, Block-Wise and Block-Bordered Magic and Bimagic Squares With Magic Sums
21, 212 and 2021. Zenodo, December 16, 2020, pp. 1-118, http://doi.org/10.5281/zenodo.4380343. [22] Inder J. Taneja, Block-Wise and Block-Bordered Magic and Bimagic Squares of Orders 10 to
47. Zenodo, January 14, 2021, pp. 1-185, http://doi.org/10.5281/zenodo.4437783. [23] Inder J. Taneja, Bordered and Block-Wise Bordered Magic Squares: Odd Order Multiples,
Zenodo, Feburary 10, 2021, pp. 1-75, http://doi.org/10.5281/zenodo.4527739 [24] Inder J. Taneja, Bordered and Block-Wise Bordered Magic Squares: Even Order Multiples,
Zenodo, Feburary 10, 2021, pp. 1-96, http://doi.org/10.5281/zenodo.4527746 • Block-Bordered Magic Squares [18] Inder J. Taneja, Block-Bordered Magic Squares of Prime and Double Prime Numbers - I, Zen-
odo, August 18, 2020, pp. 1-81, http://doi.org/10.5281/zenodo.3990291. [19] Inder J. Taneja, Block-Bordered Magic Squares of Prime and Double Prime Numbers - II, Zen-
odo, August 18, 2020, pp. 1-90, http://doi.org/10.5281/zenodo.3990293. [20] Inder J. Taneja, Block-Bordered Magic Squares of Prime and Double Prime Numbers - III,
Zenodo, September 01, 2020, pp. 1-93, http://doi.org/10.5281/zenodo.4011213. 176 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtane • Block-Wise Bordered Magic Squares [25] Inder J. Taneja, Block-Wise Bordered and Pandiagonal Magic Squares Multiples of 4, Zenodo,
August 31, 2021, pp. 1-148, https://doi.org/10.5281/zenodo.5347897. [26] Inder J. Taneja, Block-Wise Bordered Magic Squares Multiples of Magic and Bordered Magic
Squares of Order 6, Zenodo, September 10, pp. 1-99 https://doi.org/10.5281/zenodo.5500134. [27] Inder J. Taneja, Block-Wise Bordered Magic Squares Multiples of 8, Zenodo, September 17,
pp. 1-80, https://doi.org/10.5281/zenodo.5514396. [28] Inder J. Taneja, Block-Wise Bordered Magic Squares Multiples of 10, Zenodo, September 17,
pp. 1-170, https://doi.org/10.5281/zenodo.5514398. [29] Inder J. Taneja, Block-Wise Bordered and Pandiagonal Magic Squares Multiples of 12, Zenodo,
September 23, pp. 1-170, https://doi.org/10.5281/zenodo.5523608. [30] Inder J. Taneja, Block-Wise Bordered Magic Squares Multiples of 14, Zenodo, September 26,
pp. 1-198, https://doi.org/10.5281/zenodo.5528867. 177 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja
https://inderjtane [41] Inder
J.
Taneja,
Figured
Magic
Squares
of
Order
24
Using
Bordered
Magic
Rectangles:
A
Systematic
Procedure,
Zenodo,
November
29,
2022,
pp.
1-104,
https://doi.org/10.5281/zenodo.7377779. [41] Inder
J. Taneja,
Figured
Magic
Squares
of
Order
24
Using
Bordered
Magic
Rectangles:
A
Systematic
Procedure,
Zenodo,
November
29,
2022,
pp. 1-104,
https://doi.org/10.5281/zenodo.7377779. [42] Inder
J. Taneja,
Figured
Magic
Squares
of
Order
26
Using
Bordered
Magic
Rectangles:
A
Systematic
Procedure,
Zenodo,
November
29,
2022,
pp. 1-88,
https://doi.org/10.5281/zenodo.7377794. [43] Inder
J. Taneja,
Figured
Magic
Squares
of
Order
28
Using
Bordered
Magic
Rectangles:
A
Systematic
Procedure,
Zenodo,
December
02,
2022,
pp. 1-179,
https://doi.org/10.5281/zenodo.7390666. [44] Inder
J. Taneja,
Figured
Magic
Squares
of
Order
30
Using
Bordered
Magic
Rectangles:
A
Systematic
Procedure,
Zenodo,
December
02,
2022,
pp. 1-179,
https://doi.org/10.5281/zenodo.7390705. • Bordered Magic Rectangles [31] Inder J. Taneja, Different Styles of Magic Squares of Orders 6, 8, 10 and 12 Using Bordered
Magic Rectangles, Zenodo, November 14, 2022, pp. 1-26, https://doi.org/10.5281/zenodo.7319985. [32] Inder J. Taneja, Different Styles of Magic Squares of Order 14 Using Bordered Magic Rectan-
gles, Zenodo, November 14, 2022, pp. 1-40, https://doi.org/10.5281/zenodo.7319787. [33] Inder J. Taneja, Different Styles of Magic Squares of Order 16 Using Bordered Magic Rectan-
gles, Zenodo, November 14, 2022, pp. 1-63, https://doi.org/10.5281/zenodo.7320116. [34] Inder J. Taneja, Different Styles of Magic Squares of Order 18 Using Bordered Magic Rectan-
gles, Zenodo, November 14, 2022, pp. 1-85, https://doi.org/10.5281/zenodo.7320131. [35] Inder J. Taneja, Different Styles of Magic Squares of Order 20 Using Bordered Magic Rectan-
gles, Zenodo, November 14, 2022, pp. 1-88, https://doi.org/10.5281/zenodo.7320877. [36] Inder J. Taneja, Few Examples of Magic Squares of Even Orders 6 to 18 Using Bordered Magic
Rectangles, Zenodo, October 19, 2022, pp. 1-30, https://doi.org/10.5281/zenodo.7225854. [37] Inder J. Taneja, Few Examples of Magic Squares of Even Orders 20 to 30 Using Bordered Magic
Rectangles, Zenodo, October 19, 2022, pp. 1-100, https://doi.org/10.5281/zenodo.7225886. • Figured Magic Squares and Bordered Magic Rectangles [38] Inder J. Taneja, Figured Magic Squares of Orders 6, 10, 12, 14 and 16 Using Bor-
dered Magic Rectangles: A Systematic Procedure, Zenodo, November 29, 2022, pp. 1-31,
https://doi.org/10.5281/zenodo.7377674. [39] Inder
J. Taneja,
Figured
Magic
Squares
of
Orders
18
and
20
Using
Bordered
Magic Rectangles:
A Systematic Procedure, Zenodo, November 29, 2022, pp. 1-87,
https://doi.org/10.5281/zenodo.7377689. [39] Inder
J. Taneja,
Figured
Magic
Squares
of
Orders
18
and
20
Using
Bordered
Magic Rectangles:
A Systematic Procedure, Zenodo, November 29, 2022, pp. 1-87,
https://doi.org/10.5281/zenodo.7377689. 178 Figured Magic Squares of Order 30 Using Bordered Magic Rectangles: A Systematic
Procedure, Zenodo, December 02, 2022, pp. 1-179, https://doi.org/10.5281/zenodo.7390705 Inder J. Taneja
https://inderjtaneja.com; https://numbers-magic.com; Inder J. Taneja [40] Inder
J. Taneja,
Figured
Magic
Squares
of
Order
22
Using
Bordered
Magic
Rectangles:
A
Systematic
Procedure,
Zenodo,
November
29,
2022,
pp. 1-61,
https://doi.org/10.5281/zenodo.7377706. • Creative Magic Squares [45] Inder J. Taneja, Creative Magic Squares: Area Representations, Zenodo, June 22, pp. 1-45,
2021, http://doi.org/10.5281/zenodo.5009224. 179 179
| 39,597
|
2016LORR0090_7
|
French-Science-Pile
|
Open Science
|
Various open science
| 2,016
|
Comportement inclusionnaire dans un bain d’aluminium brassé par induction
|
None
|
English
|
Spoken
| 5,143
| 8,312
|
[82] E. Pauty, B. Laboudigue and J. Etay, "Numerical simulation of the flow and the solid transport when tilting a holding furnace," Metallurgical and Materials Transactions B, vol. 31B, p. 207, 2000. [83] S. Instone, A. Buchholz and G. Gruen, "Inclusion Transport Phenomena in Casting Furnaces," Light Metals 2008, p. 811, 2008. [84] L. Nastac, D. Zhang, S. Jia and Y. Xuan, "Advances on Experimental and Numerical Modeling of Al-based Alloys and Nanocomposites Fabricated via Ultrasonic and Electromagnetic Processing," in Proceedings of EPM2015, 8th International Conference on Electromagnetic Processing of Materials, Cannes, 2015. [85] D. Shu, J. Wang and B. Sun, "Online Electromagnetic Filtration of Molten Aluminum Using a Multistage Separator System," Journal for Manufacturing Science and Production, vol. 15, no. 1, pp. 89-92, 2015. [86] M. W. J. A. B. a. R. E. A. Kennedy, "Impact of Coil Geometry on Magnetohydrodynamic Flow in Liquid Aluminium and Its Relevance to Inclusion Separation by Electromagnetophoresis," Journal for Manufacturing Science and Production, vol. 15, no. 1, pp. 69-78, 2015. [87] S. Shimasaki and S. Taniguchi, "Separation Efficiency of Inclusion Particles from Liquid Aluminum by Electromagnetic Cyclone Separator," in Proceedings of EPM2015, 8th International Conference on Electromagnetic Processing of Materials, Cannes, 2015. [88] L. Claudotte, "Prédiction de la taille et de la composition des inclusions dans une poche d'acier liquide: étude numérique (PhD Dissertation)," Université de Lorraine, Nancy, 2010. [89] J. P. Bellot, V. De Felice, B. Dussoubs, A. Jardy and S. Hans, "Coupling of CFD and PBE Calculations to Simulate the Behavior of an Inclusion Population in a Gas-Stirring Ladle," Metallurgical and Materials Transactions B, vol. 45B, p. 13, 2014. [90] Q. Wang, F. Qi, B. Li and F. Tsukihashi, "Behavior of Non-metallic Inclusions in a Continuous Casting Tundish with Channel Type Induction Heating," ISIJ International, vol. 54, no. 12, pp. 2796-2805, 2014. [91] E. Waz, A. Bansal, P. Chapelle, Y. Delannoy, J. P. Bellot and P. L. Brun, "Modeling of Inclusion Behavior in an Aluminum Induction Furnace," in Light Metals 2016, John Wiley & Sons, 2016. [92] D. Leenov and A. Kolin, "Theory of electromagnetophoresis. I. Magnetohydrodynamic forces experienced by spherical and symmetrically oriented cylindrical particles," The Journal of Chemical Physics, vol. 22, no. 4, p. 683, 1954. [93] Z. Sun, M. Guo, J. Vleugels, O. Van der Biest and B. Blanpain, "Numerical calculations on inclusion removal from liquid metals under strong magnetic fields," Progress In Electromagnetics Research, vol. 98, p. 359, 2009. [94] S. Taniguchi and J. K. Brimacombe, "Application of pinch force to the separation of inclusion particles from liquid steel," ISIJ international, vol. 34, no. 9, p. 722, 1994. [95] L. Zhang, S. Wang, A. Dong, J. Gao and L. N. W. Damoah, "Application of Electromagnetic (EM) Separation Technology to Metal Refining Processes: A Review," Metallurgical and Materials Transactions B, vol. 45B, p. 2153, 2014. [96] R. Bolcato, J. Etay, Y. Fautrelle and H. K. Moffatt, "Electromagnetic billiards," Physics of Fluids A: Fluid Dynamics, vol. 5, no. 7, p. 1852, 1993. [97] N. El-Kaddah, A. D. Patel and T. T. Natarajan, "The electromagnetic filtration of molten aluminum using an induced-current separator," JOM, vol. 47, no. 5, p. 46, 1995. [98] P. er and J. Driole, "Effects of the electromagnetic stirring on the removal of inclusions of oxide from liquid steel," Metallurgical Transactions B, vol. 13B, p. 45, 1982. [99] D. R. Sadoway and J. Szekely, "A new experimental technique for the study of turbulent electromagnetically driven flows," Metallurgical and Materials Transactions B, vol. 11B, p. 334, 1980. [100] M. Ščepanskis, K. Thomsen, A. Jakovičs, T. Beinerts, M. Sarma, R. Nikoluškins and A. Bojarevičs, "How to make liquid metal transparent? A neutron radiography method for scaled down metallurgical equipment," in Proceedings of the 1st Thermal and Fluid Engineering Summer Conference, New York, 2015. [101] O. J. Ilegbusi and J. Szekely, "On the flow criteria for suspending solid particles in inductively stirred melts: Part I. newtonian behavior," Metallurgical Transactions B, vol. 19B, p. 557, 1988. 160 [102] V. Bojarevics, K. Pericleous and R. Brooks, "Dynamic model for metal cleanness evaluation by melting in a cold crucible," Metallurgical and Materials Transactions B, vol. 40B, p. 328, 2009. [103] K. A. Pericleous and S. N. Drake, "An algebraic slip model of PHOENICS for multi-phase applications," in Numerical simulation of fluid flow and heat/mass transfer processes, Springer Berlin Heidelberg, 1986, p. 375. [104] M. Cross, T. N. Croft, G. Djambazov and K. Pericleous, "Computational modelling of bubbles, droplets and particles in metals reduction and refining," Applied mathematical modelling, vol. 30, no. 11, p. 1445, 2006. [105] K. Takahashi and S. Taniguchi, "Electromagnetic separation of nonmetallic inclusion from liquid metal by imposition of high frequency magnetic field," ISIJ international, vol. 43, no. 6, p. 820, 2003. [106] S. Wang, L. Zhang, Y. Tian, Y. Li and H. Ling, "Separation of non-metallic inclusions from molten steel using high frequency electromagnetic fields," Metallurgical and Materials Transactions B, vol. 45 , p. 1915, 2014. [107] C. T. Crowe, J. D. Schwarzkopf, M. Sommerfeld and Y. Tsuji, Multiphase Flows with Droplets and Particles, CRC Press, 1997. [108] M. 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Signorino, G. Baldi, A. A. Barresi, M. Cournil, J. J. Derksen, F. Gruy, E. Hollander, M. Kraume, D. Marchisio, A. Paschedag, M. Vanni, H. E. A. Van den Akker and A. Varone, "Agglomeration in Pratsolis Project". [115] P. Saffman and J. S. Turner, "On the collision of drops in turbulent clouds," Journal of Fluid Mechanics, vol. 1, no. 1, p. 16, 1956. [116] J. Abrahamson, "Collision rates of small particles in a vigorously turbulent fluid," Chemical Engineering Science, vol. 30, no. 11, p. 1371, 1975. [117] L. I. Zaichik, O. Simonin and V. M. Alipchenkov, "Turbulent collision rates of arbitrary-density particles," International Journal Heat and Mass Transfer, vol. 53, no. 9, p. 1613, 2010. [118] M. Soos, L. Wang, R. O. Fox, J. Sefcik and M. Morbidelli, "Population balance modeling of aggregation and breakage in turbulent Taylor–Couette flow," Journal of Colloid and Interface Science, vol. 307, no. 2, p. 433, 2007. [119] D. Ramkrishna, Population balances: Theory and applications to particulate systems in engineering, Academic press, 2000. [120] D. Ramkrishna and M. R. Singh, "Population balance modeling: Current status and future prospects," Annual Review of Chemical and Biomolecular Engineering, vol. 5, p. 123, 2014. [121] D. L. Marchisio, R. D. Vigil and R. O. Fox, "Quadrature method of moments for aggregation–breakage processes," Journal of Colloid and Interface Science, vol. 258, no. 2, p. 322, 2003. [122] D. L. Marchisio and R. O. Fox, "Solution of population balance equations using the direct quadrature method of moments," Journal of Aerosol Science, vol. 36, no. 1, p. 43, 2005. [123] S. Kumar and D. Ramkrishna, "On the solution of population balance equations by discretization—I. A fixed pivot technique," Chemical Engineering Science, vol. 51, no. 8, p. 1311, 1996. [124] J. Kumar, M. Peglow, G. Warnecke, S. Heinrich and L. Mörl, "Improved accuracy and convergence of discretized population balance for aggregation: The cell average technique," Chemical Engineering Science, vol. 61, no. 10, p. 3327, 2006. [125] F. Anker, S. Ganesan, V. John and E. Schmeyer, "A comparative study of a direct discretization and an operator-splitting solver for population balance systems," Computers & Chemical Engineering, vol. 75, p. 95, 2015. [126] M. Smoluchowski, "Versuch einer mathematischen Theorie der Koagulationskinetik kolloiderLösnngen," Zeitschrift fuer physikalische Chemie, vol. 92, p. 129, 1917. [127] Z. Popovic and B. Popovic, Introductory Electromagnetics, New Jersey: Prentice Hall, 1999. [128] V. Bojarevics, R. Harding, K. Pericleous and M. Wickins, "The development and experimental validation of a numerical model of an induction skull melting furnace," Metallurgical and Materials Transactions B, vol. 35B, p. 785, 2004. 161 [129] V. V. Yashchuk, S. Lee and E. Paperno, "Magnetic shielding," in Optical Magnetometry, 1st ed., D. Budker and D. Kimball, Eds., Cambridge University Press, 2013, p. 225. [130] M. Zucca, G. Crotti, O. Bottauscio, X. Li, M. Di Pardo and M. Chiampi, "Three-dimensional modeling for magnetic field shielding in a high electric power process," Journal of Applied Physics, vol. 99, no. 8, p. 08P503, 2006. [131] D. van Riesen, G. Henneberger and C. Kaehler, "Comparison of different formulations for eddy-current computations in an induction furnace," Progress in Electromagnetic Research Symp, p. 623, 2004. [132] G. Henneberger, P. K. Sattler, D. Shen and W. Hadrys, "Coupling of magnetic and fluid flow problems and its application in induction melting apparatus," IEEE Transactions on Magnetics, vol. 29, no. 2, p. 1589, 1993. [133] A. Kumar, S. K. Ajmani and A. R. Pal, "Optimal configuration of blast furnace slag runner to reduce fluid flow stresses at wall using mathematical modelling," Ironmaking and Steelmaking, vol. 37, no. 1, p. 9, 2010. [134] M. Kostoglou and A. Karabelas, "On the self-similarity of the aggregation–fragmentation equilibrium particle size distribution," Journal of Aerosol Science, vol. 30, no. 2, p. 157, 1999. [135] R. Vigil, "On equilibrium solutions of aggregation–fragmentation problems," Journal of Colloid and Interface Science, vol. 336, no. 2, p. 642, 2009. [136] M. Ščepanskis, "The Modelling of the Behaviour of Solid Inclusions in the EM Induced Recirculate Turbulent Flows of Liquid Metal (PhD Dissertation)," University of Latvia, Riga, 2014. [137] J. Ngoma and J. Kroll-Rabotin, "Numerical study of aggregation dynamics with a Lattice Boltzmann Method coupled with discrete elements through flow penalization," in International Conference on Multiphase Flow, Florence, 2016. [138] H. Gaye, C. Gatellier, M. Nadif, P. Riboud, J. Saleil and M. Faral, "Slags and inclusions control in secondary steelmaking," Revue de Métallurgie, vol. 85, p. 759, 1987. [139] J. Szekely, T. Lehner and C. Chang, "Flow phenomena,mixing and mass transfer in argon-stirred ladles," Ironmaking and Steelmaking, vol. 6, p. 285, 1979. [140] H. Gaye, J. Lehmann, P. Rocabois and F. Ruby-Meyer, "Computational thermodynamics and slag modelling applied to steel elaboration," Steel Research, vol. 72, p. 446, 2001. [141] M. Hallberg, P. Jönsson, T. Jonsson and R. Erikson, "Process model of inclusion separation in a stirred steel ladle," Scandinavian Journal of Metallurgy, vol. 34, p. 41, 2005. [142] J. Aoki, B. Thomas, J. Peter and K. Peaslee, "Experimental and theoretical investigation of mixing in a bottom gas-stirred ladle," in AISTech, 2004. [143] L. Zhang, "Transport phenomena and CFD application during process metallurgy," in Advanced Processing of Metals and Materials: Iron and Steel Making, 2006. [144] T. Kato, S. S. Shimasaki and S. S. Taniguchi, "Water model experiments for hydrodynamics forces acting on inclusion particles in molten metal under turbulent conditions," in Jim Evans Honorary Symposium, TMS 2010, 2010. [145] V. Warke, S. Shandar and M. Makhlouf, "Mathematical modelling and computer simulation of molten aluminium cleaning by the rotating impeller degasser," Journal of Materials Processing Technology, vol. 168, 2005. [146] A. Bansal, P. Chapelle, Y. Delannoy, E. Waz, . Le Brun and J. Bellot, "Experimental and Numerical Analysis of the Deformation of a Liquid Aluminum Free Surface Covered by an Oxide Layer During Induction Melting," Metallurgical and Materials Transactions B, vol. 46, p. 2096, 2015. [147] The Aluminum Association, "International Alloy Designations and Chemical Composition Limits for Wrought Aluminum and Wrought Aluminum Alloys," The Aluminum Association, 2009. [148] O. Ubbink, "Numerical prediction of two fluid systems with sharp interfaces (PhD Dissertation)," Imperial College of Science, London, 1997. [149] D. Youngs, "Time dependent multi-material flow with large fluid distortion," Numerical Methods for Fluid Dynamics, p. 273, 1982. 162 Annex Annex 1: Composition and properties of Aluminum alloys
1050 2024 5182 7050 Si 0.25 0.15 0.2 0.12 Fe 0.4 0.2 0.35 0.15 Table 25: Composition of aluminum alloys [147] Cu Mn Mg Cr Ni Zn 0.05 0.05 0.05...... 0.07 3.7-4.5 0.15-0.8 1.2-1.5 0.1... 0.25 0.15 0.20-0.50 4.0-5.0 0.1... 0.25 1.7-2.4 0.04 1.7-2.6 0.04 0.03 5.7-6.9 Ti 0.05 0.15 0.1 0.06 Zr......... 0.05-0.12 Table 26: Physical properties of aluminum alloys Alloys 1050 T (°C) 660 700 1000 Law f(T) 2024 5182 7050 750 700 700 Dynamic Electrical conductivity Density [62] viscosity [63] [62] (kg.m-3) (Pa.s) (-1.m-1) 2390 1.28E-03 4.04E+06 2379 1.15E-03 3.97E+06 2339 7.10E-04 3.33E+06 = − = 6 0.15 − 5.91x10 − 2.03x103T 2416 1.04E-03 3.38E+06 2337 3.53E+06 1.15E-03 2522 3.77E+06 Surface tension coefficient [57] (N.m-1) 0.868 0.865 0.783 = − -3 0.25x10 − 0.795 0.860
163 Annex 2: Experimental techniques for free surface deformation study
Table 27: Summary of experimental techniques available for the measurement of a deformable surface Technique Strengths Limitations Easy application Limited precision Varied range Laser telemetry Single point measurement Good data acquisition rate High cost Interferometry Very high precision Reflective surfaces only Laser triangulation Simple and accurate Independent of illumination High data acquisition rate Structured light Accurate pattern projection High data acquisition rate Photogrammetry and Stereoscopy Electrical potential probe Thermocouple based contact probe Simple application Good accuracy Easy application Laser safety Reconstruction of image – algorithm required Reconstruction of data images Algorithms required Lower data acquisition rate Interference with EM field Intrusive and low temperature range Intrusive Thermocouple response time
164 Annex 3: Modeling of a free surface: Emphasis on VOF discretization techniques
Free surface flows require an accurate representation of the shape and the location of the free surface as well as its temporal evolution. For an arbitrary Eulerian mesh, a variety of numerical methods to calculate free surface deformation exists. These can be broadly categorized into Surface or Volume methods [19]. In Surface methods the interface is either explicitly tracked by marking it with special marker points or by attaching the interface to the mesh surface. On the other hand, in Volume methods, the fluids on either side of the interface are marked by particles (markers in fluid) of negligible mass or by an indicator function. We focus on the VOF method, which is quite economical as only one value (i.e. the volume fraction of a given phase) must be stored in each mesh element. Yeoh and Tu [19] and Ubbink [148] can be referred for further VOF method related information. VOF discretization techniques can be broadly categorized into two groups: Donor–Acceptor formulation and line techniques. D-A formulation basic idea is to exploit the information about the volume fraction downstream (i.e. Acceptor cell) as well as upstream (i.e. Donor cell) of a cell face in order to establish a crude interface shape and then to employ this shape while computing the flux across that face. Line techniques on the other hand fit the interface using oblique lines or piecewise linear segments. A comparison of different line techniques is illustrated in Figure 102. The Piecewise Linear Interface Construction (PLIC) technique proposed by Youngs [149] is an example. A major drawback of this technique is the discontinuities that it gives when the interface is reconstructed using ‘unjoined’ segments.
Figure 102: Comparison of different line techniques [148] Concerning VOF method, Table 28 compares some interface advection schemes available in ANSYS Fluent.
165 Table 28:
VOF
ad
vection schemes [73] Scheme Implicit Explicit Accuracy Computational time QUICK YES YES Low High Modified HRIC YES YES Medium High CICSAM NO YES High Medium Compressive YES YES High Medium to High GeoReconstruct NO YES Very High Low to Medium BGM YES NO Very High Low to Medium GeoReconstruct has been selected for the present study as this scheme provides the best compromise for calculation of a sharp interface while limiting computing resources. GeoReconstruct is based on the PLIC method and can be described in the following manner: Step1: calculation of a linear projection of the interface in each cell containing an interface, with the help of the volume fraction and its gradient. Step2: calculation of the advection of fluid through each face with the help of the calculated linear interface (step 1) as well as velocity distribution on the faces. Step3: calculation of the new volume fraction value in each cell using flux balancing calculated in step 2.
166 Annex 4: Interfacial area density approximation method
As previously presented in A3.2.1.2, the interfacial area density can be approximated by three different expressions [73] [74]. Since the precision of these three approximations is unknown, we have performed tests with known interface shapes. Interface shapes such as those of a cylinder, a hollow cylinder, a sphere and a torus for which the exact surface area value may be obtained analytically were successively considered. The interfacial area density calculated using the expressions in Table 8 was integrated over the complete shape volume to obtain the total surface area S. Table 29 presents a comparative analysis of the analytical area value with that calculated by the various expressions. For cylindrical shapes, A is the most precise expression, while for a simple spherical interface approximation B gives better results and for a torus approximation C is best suited. Thus, it can be concluded that the best approximation should be selected according to the case under study. Table 29: Relative error associated to various interfacial area approximation methods
Axis of symmetry Shape 1) Cylinder 1 4 2) Hollow cylinder 3 2 3) Sphere 4) Torus SA =| -0,39% | SB = | -2,62% | = SC − 10,72% -0,35% -3,37% 3,91% 8,30% 1,61% -10,92% 15,83% 10,23% 0,93% | | * error % calculated with respect to Stheory
Similar tests were performed for the case of a dome shaped interface, representative of the deformation of the liquid metal free surface encountered in an induction furnace. This interface was fitted with a polynomial of degree six. The polynomial equation allowed us then to obtain by numerical integration a theoretical value of the surface area, which was then compared with the integral of the interfacial area density obtained using the various expressions, as presented in Table 30. A conclusion can be drawn in the favor of approximation C.
Table 30: Comparison of various expressions to calculate the area of a dome interface Dome shaped interface SA SB SC 25% 33% 10% * error % calculated with respect to Stheory,fitted 167
Annex 5: Particle size distribution function for MgAl2O4
The characteristics of the non-metallic and undissolved inclusions (type, density, size and shape) considered in the present work originate from measurements by LiMCA and PoDFA techniques. A database was developed at C-TEC, describing several characteristics for each type of inclusion: density, minimum, mean and maximum diameters, diameter distribution, shape and a frequency index of observation in PoDFA samples. Thanks to these observations, it was possible to reconstruct for MgAl2O4 inclusions (spherical shape) a representative particle size distribution (Figure 103–black curve). This distribution was fitted using a log-normal distribution [91] (Figure 103– red curve), as expressed in Eq. 110. (, 300 )=. , e p (− ( (, Ni (#) 250 )−. ) (110) ) Ni (#) Ni (log-normale)
200
150 100 50 0 -50 0 50 100 150 200 250 300 350 dd(microns) p (μm) p
Figure 103: Particle size distribution measured and fitted (using log-normal law)
This deduced log-normal law was adopted as the initial inclusion psd while calculating the inclusion behavior in the aluminum bath in chapter C2.
168 Annex 6: Total turbulent collision rate
Zaichik et al. [117] presented the turbulent collision rate between two particles as a sum of and. The former represents the inertial collision rate while the latter corresponds to the spatial collision rate. =
( =( − =| − + + + | ) − ) − / (111) / ′ (112
)
/ +
(113) ′, and respectively represent the velocity fluctuation of the fluid, the response coefficient and the longitudinal structure function. The response coefficient given by Eq. 114 is a function of and μ ≡ ⁄, which are in turn functions of the particle response time,, the ≡,⁄ eddy particle interaction time scale, the Taylor differential timescale and the Lagrangian integral timescale.
= + + (114) +
In Eq. 115, within the viscous range (, ), the longitudinal structure function solely depends on the turbulence dissipation rate, the kinematic viscosity and the collision diameter. Over the remaining spatial range, the function may be written as a function of the velocity fluctuation, the dimensionless collision diameter ̅ =, ⁄, the Taylor scale Reynolds number = ′ ⁄ / = { and a constant ′ =. ( − e p − ̅, / ) ; /, ⁄
In the above expressions, = + + the densities of the fluid and the particle phases, where equal to 0.5 for spherical particles. ̅ + ̅ / / (115) ; corresponds to the ratio between is the added mass coefficient and is
169
Annex 7: Oxide layer fragmentation observations in a lab scale IMF
The breaking-up of oxide layers was observed during some experiments at the industrial scale. Therefore, in order to reproduce and study this phenomenon at the lab scale, qualitative observations (camera images and videos) were made during the structured light campaign, where the filling levels, the electrical parameters and the alloy type were varied. In the case of Al-1050, only slight stretching of the oxide layer was observed without any breaking. This was also observed for Al-7050, where the stretching increased as a function of the generator power. An illustration of such a behavior for Experiment 14 is seen in Figure 104. The white lines all across the free surface are due to the oxide layer stretching, without any visual evidence of fragmentation. We also observe two zones where the liquid metal seems stuck to the refractory wall, which could be an unintended result of the melting process of solid blocks of metal alloy.
Figure 104: Camera image of oxide layer stretching (Al-7050 at 50 % filling level and Pgen = 15 kW)
On the other hand, due to the presence of highly reactive magnesium, oxide layers for alloy Al5182 behaved differently. Figure 105(a – c) presents the raw images for a varying generator power. A top layer of crumbled oxides was observed, while any, if at all present, oxide layer fragmentation was hidden underneath. The crumbled oxides may be attributed to the presence of a large quantity of magnesium oxides. (a) P gen = 10 kW (
b) P gen = 15 kW (c) P gen = 20 kW Figure 105: Raw images for different generator powers for Al-5182 (75 % filling) Experiment 15 was performed, where the previously observed crumbled layer was carefully skimmed off and revealed the underlying oxide layer.
Consequently
,
slight oxide layer stretch marks were observed, quite similar to previously noted Al-7050 layer
. Additionally, we also used 170 a high speed camera to locally study any breaking and re-oxidation between the stretched zones. However, apart from small free surface fluctuations, no other discernable observation could be made. The small crucible diameter (~10 cm) and the ensuing wall effect on the forces acting on the oxide layer could play a role in such a behavior of the oxide layer. Thus for the current configuration, we did not observe any oxide layer fragmentation. The experiments at the industrial scale, on the other hand, show evidence of some fragmentation of the oxide layer which could not be studied in the scope of this dissertation.
171 Annex 8: Free surface deformation measurements by laser telemetry in a lab scale
IMF
The Time of Flight (ToF) technique, described in section A.2.2.1, was implemented using a laser telemetry device (Figure 106 (a)). A Bosch GLM250VF laser telemeter was used on a support system which allowed two degrees of freedom along two orthogonal horizontal axes (Figure 106 (b)) with a step of 10 mm. Figure 106: (a) Laser Telemeter setup over the inductor, (b) x and y locations of the telemetry measurements
To avoid overheating of the device, the generator power was cut off between each point of measurement, so that the method follows the sequence in Figure 107. Measure
with telemeter Power switch off Displace the telemeter Power switch on
Figure 107: Laser telemeter measurement protocol
Distance between the dome surface and the telemeter was measured. The distance of the initial free surface (at Pgen = 0 kW) was measured and taken as the reference distance. These two distances approximated the relative deformation of the free surface as a function of the operating parameters. 172
Figure 108 shows the deformation along the two axes in 3D for Experiment 3 (75% liquid metal filling and 20 kW generator power).
The
deformation of the free surface along the y axis was
±
25
mm
, while
that
along the x axis was between -40
mm
and +25 mm, with respect to the level (z=0 mm). Figure 108: 3D representation of the free surface deformation measured by laser telemetry – Experiment 3
It can be observed that the only intersecting point at x = y = 0 (encircled in dotted black), did not have the same deformation. This is due to the experimental protocol where the generator power was switched off during every telemeter displacement. In some cases, this made the oxide layer stick to the crucible walls which disturbed the surface deformation and thus rendered the results non-reproducible. Therefore, the use of laser telemetry to measure the spatial as well as the temporal evolution of the free surface was deemed impractical. 173 Annex 9: Impact of the turbulence damping factor near the bath free surface
The need of turbulence damping at the interfacial region has been discussed in section A.3.2.1.1. The additional turbulence damping source term in the ω equation is written as:, = ∆ ( ∆ ) (116) B is the turbulence damping factor. This adjustable factor was previously kept at its default value, equal to 10 as proposed in the ANSYS Fluent documentation. Since damping is active only in the interfacial region, it was hypothesized that any modification of the turbulence damping factor may impact the free surface deformation. Therefore, simulations while varying the damping factor were performed. Figure 109 presents the dome profile for B = 0 and B = 100.
Figure 109: Comparison of dome profiles: B = 0 vs B = 100 – Experiment 9
It is noted that the variation in the turbulence damping factor does not have any significant impact on the dome profile. 174 Résumé en Français
Dans le secteur aéronautique, la performance des alliages d’aluminium connait une amélioration continue, grâce notamment à l’optimisation des procédés d’élaboration. Dans ce cadre, le travail de recherche vise à prédire le comportement des inclusions dans un bain d’aluminium brassé par induction afin d’améliorer la propreté inclusionnaire des alliages coulés. Un modèle numérique a été développé pour simuler le comportement magnétohydrodynamique du bain d’aluminium dans le creuset suivi par la modélisation du comportement d’une population d’inclusions non-métalliques. Le modèle 2D axisymétrique en régime transitoire s’appuie sur le code de CFD commercial ANSYS Fluent, bien que de nombreuses fonctions utilisateurs aient été introduites pour simuler les phénomènes spécifiques comme l’induction électromagnétique et la résolution des bilans de population. Le modèle MHD résout dans un unique maillage les phénomènes d’induction électromagnétiques, l’écoulement turbulent du bain d’Al, la déformation de la surface libre et les effets de la présence d’une couche de métal oxydée en surface du bain. Une méthode dite de vitesse de glissement (entre les particules et le fluide) a été choisie pour simuler à la fois le transport macroscopique des inclusions dans le bain d’Al et les interactions mésoscopique entre les inclusions (c.à.d. les mécanismes d’agrégation et de fragmentation). Des campagnes expérimentales à l’échelle d’un four de laboratoire et d’une installation industrielle accompagnent le travail numérique pour le valider. Les résultats de modélisation MHD exprimés sous la forme du profil de déformation du bain sont en accord raisonnable avec les mesures faites au laboratoire. Les résultats numériques démontrent également l'effet du frottement induit par la couche d'oxyde sur le profil du bain, a que sur l'écoulement à proximité de la surface du dôme. Pour des conditions opératoires du four industriel en mode de maintien, l’évolution temporelle de la population au sein du bain est calculée. Il apparaît que la séparation magnétique est très intense, particulièrement dans la peau électromagnétique, et est ainsi responsable du transport et de la capture d’une grande fraction de la population d’inclusions à la paroi du four. Mots clés : Inclusions, Ecoulement Multiphasique, Alliage Aluminium, Mécanique des Fluides Numérique, Métrologie Abstract
With an objective of improving processing and development of aerospace aluminum alloys, the current dissertation presents experimental and numerical tools which help comprehend the behavior of a non-metallic inclusion population in an Al bath stirred by induction. The mechanisms occurring in the metallurgical reactor were separated into two interlinked issues – (i) Magnetohydrodynamics (MHD) of the induction furnace, and (ii) Inclusion population dynamics in the Al bath, which were modeled using the ANSYS Fluent software and in-house User Defined Functions. For a 2D axisymmetric geometry, numerical simulations were performed in a single framework and calculated: (i) the electromagnetic forces using the A-V formulation, (ii) the free surface deformation using the Volume Of Fluid method, (iii) the turbulent stirring of the bath using a RANS-based k-omega model and (iv) the friction force due to the oxide layer by imposing a pseudo-wall condition on the bath free surface. The steady state MHD results and the physical properties of the inclusion population were used as input data for the transient inclusion behavior modeling. A combination of the Drift Concentration Method and the Population Balance Method was developed to respectively model the mean transport of inclusions within the bath at the macroscopic scale and the inclusion interactions (turbulent aggregation and fragmentation) at the mesoscopic scale. The performance of the MHD numerical tool was evaluated by comparing the model results with experimental results at laboratory and industrial scales. The simulation results in the form of the average bath surface profile were found to be consistent with the laboratory measurements. The results also illustrated the impact of the friction due to the oxide layer on the bath surface deformation as well as on the flow near the dome interface. The inclusion behavior simulations were performed for the holding mode operation of an industrial IMF. The deduced removal frequency compared the relative importance of each phenomeno . It was found that the electromagnetic migration, especially in the electromagnetic skin, dominates the inclusion dynamics and is responsible for the capture of a large fraction of the inclusion population. Keywords: Inclusions, Multiphase flow, Aluminum alloys, Computational Fluid Dynamics, Experimental analysis 175.
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¿SON LOS TÉS E INFUSIONES TAN BENEFICIOSOS COMO NOS CUENTAN? DESCUBRE EL MUNDO DE LOS ALCALOIDES
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¿SON LOS TÉS E INFUSIONES TAN BENEFICIOSOS COMO NOS
CUENTAN? DESCUBRE EL MUNDO DE LOS ALCALOIDES
Begoña Fernández-Pintor, Lorena González-Gómez, Judith Gañán, Natalia Casado,
Sonia Morante-Zarcero, Damián Pérez-Quintanilla e Isabel Sierra.
Universidad Rey Juan Carlos, Escuela superior de Ciencias y Tecnología, Departamento de
Tecnología Química y Ambiental (begona.fernandez@urjc.es; lorena.gonzalez@urjc.es)
Palabras clave: alcaloides tropánicos; alcaloides pirrolizidínicos; seguridad alimentaria;
infusiones; análisis.
¿Son seguros los alimentos que comemos? Esta pregunta es muy común en la
sociedad actual dado que cada vez estamos más concienciados con la Seguridad
Alimentaria y el consumo de alimentos saludables y seguros. La Autoridad Europea
de Seguridad Alimentaria (EFSA) es el organismo encargado de la regulación de los
límites máximos o de la prohibición de ciertas sustancias que pueden resultar
perjudiciales para la salud en los alimentos. Dicho organismo se apoya en el trabajo
de multitud de científicos encargados del desarrollo de metodologías eficientes para
poder determinar dichos tóxicos y así poder establecer límites seguros en los
alimentos.
Dentro de los contaminantes químicos que aparecen en los alimentos se encuentra
una familia de tóxicos llamada toxinas naturales que son aquellas sintetizadas de
forman natural por determinadas plantas a modo de defensa contra depredadores
como pueden ser los insectos. En concreto, los alcaloides tropánicos y los alcaloides
pirrolizidínicos aparecen en nuestra dieta diaria fundamentalmente debido a una
contaminación cruzada desde plantas productoras de dichos alcaloides hasta los
alimentos que ingerimos. Algunos de los alimentos más susceptibles de contener
dichos contaminantes son de origen vegetal como los tés, las infusiones, las
especias, entre otros. Cuando ingerimos estos alimentos contaminados por
alcaloides pirrolizidínicos, se producen daños en la salud tales como daño hepático,
alteraciones respiratorias e incluso alguno de ellos pueden resultar carcinógenos
(clasificados como grupo 2B según la Agencia Internacional para la Investigación del
Cáncer) cuando se lleva a cabo un consumo prolongado de los mismos. En cambio,
si consumimos alimentos contaminados por alcaloides tropánicos, podemos
experimentar efectos como taquicardias, espasmos e incluso delirios [1,2].
Es por esto por lo que nuestro grupo de investigación se centra en el desarrollo de
metodologías que permitan la extracción y posterior análisis de alcaloides tropánicos
y pirrolizidínicos en diversas matrices alimentarias. Para ello, empleamos diferentes
técnicas y diversos equipos de laboratorio de compleja tecnología. Otro de los
puntos clave de nuestras investigaciones es el estudio de cómo afecta el tratamiento
térmico a alimentos contaminados con estos alcaloides. De tal manera, nuestro
grupo reproduce las condiciones en las que se elaboran y consumen ciertos
alimentos, como por ejemplo los tés, para evaluar la posible degradación que
pueden sufrir los alcaloides y poder estimar el riesgo de la ingesta de estos
alimentos presuntamente contaminados [3,4].
Agradecimientos
Los autores agradecen la financiación recibida del MCIU/AEI/FEDER, UE para el proyecto
RTI2018-094558-B-I00 y del MCIN/AEI /10.13039/501100011033 / FEDER, UE para el proyecto
PID2022-137278OB-I00.
Referencias
[1] L. González-Gómez, S. Morante-Zarcero, D. Pérez-Quintanilla, I. Sierra, Occurrence and
Chemistry of Tropane Alkaloids in Foods, with a Focus on Sample Analysis Methods: A Review on
Recent Trends and Technological Advances, Foods. 11 3 (2022) 407.
[2] N. Casado, S. Morante-Zarcero, I. Sierra, The concerning food safety issue of pyrrolizidine
alkaloids: An overview, Trends in Food Science & Technology. 120 (2022) 123-139.
[3] L. González-Gómez, J. Gañán, S. Morante-Zarcero, D. Pérez-Quintanilla, I. Sierra, Atropine and
scopolamine occurrence in spices and fennel infusions, Food Control. 146 (2023) 109555.
[4] B. Fernández-Pintor, N. Casado, S. Morante-Zarcero, I. Sierra, Evaluation of the thermal stability
and transfer rate of pyrrolizidine alkaloids during brewing of herbal infusions contaminated with
Echium vulgare and Senecio vulgaris weeds, Food Control, 153 (2023) 109926.
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Why is leptospirosis hard to avoid for the impoverished? Deconstructing leptospirosis transmission risk and the drivers of knowledge, attitudes, and practices in a disadvantaged community in Salvador, Brazil
|
Fabiana Almerinda G. Palma
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English
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Why is leptospirosis hard to avoid for the
impoverished? Deconstructing leptospirosis
transmission risk and the drivers of
knowledge, attitudes, and practices in a
disadvantaged community in Salvador, Brazil Fabiana Almerinda G. PalmaID1☯, Federico Costa1,2☯*, Ricardo LustosaID1, Hammed
O. MogajiID1, Daiana Santos de Oliveira2, Fa´bio Neves SouzaID3, Mitermayer G. ReisID2,4,5,
Albert I. KoID2,5, Michael BegonID6☯, Hussein KhalilID7☯ a1111111111
a1111111111
a1111111111
a1111111111
a1111111111 a1111111111
a1111111111
a1111111111
a1111111111
a1111111111 1 Federal University of Bahia, Salvador/Institute Health Collective, Salvador, Bahia, Brazil, 2 Instituto
Gonc¸alo Moniz, Fundac¸ão Oswaldo Cruz, Ministe´rio da Sau´de, Salvador, Bahia, Brazil, 3 Federal University
of Bahia/Institute of Biology, Salvador, Bahia, Brazil, 4 Faculdade de Medicina da Bahia, Federal University
of Bahia, Salvador, Brazil, 5 Department of Epidemiology of Microbial Diseases, Yale School of Public
Health, New Haven, Connecticut, United States of America, 6 Department of Evolution, Ecology and
Behaviour, The University of Liverpool, United Kingdom, 7 Department of Wildlife, Fish, and Environmental
Studies, Swedish University of Agricultural Sciences, Umeå, Sweden OPEN ACCESS ☯These authors contributed equally to this work. * fcosta2001@gmail.com ☯These authors contributed equally to this work. Citation: Palma FAG, Costa F, Lustosa R, Mogaji
HO, de Oliveira DS, Souza FN, et al. (2022) Why is
leptospirosis hard to avoid for the impoverished? Deconstructing leptospirosis transmission risk and
the drivers of knowledge, attitudes, and practices in
a disadvantaged community in Salvador, Brazil. PLOS Glob Public Health 2(12): e0000408. https://
doi.org/10.1371/journal.pgph.0000408 PLOS GLOBAL PUBLIC HEALTH PLOS GLOBAL PUBLIC HEALTH Introduction shared publicly because of personal information of
participants in the KAP survey, sero-survey, at the
individual and household and peri-domiciliary level. Researchers who wish to access the data can
contact the data manager at the Oswaldo Cruz
Foundation, Nivison Junior (nivisonjr@gmail.com). The incidence of infectious diseases in disadvantaged communities, especially in large urban
centers, is higher than incidence elsewhere [1]. There are demographic, socioeconomic, and
environmental risk factors contributing to the maintenance of such disparity [2]. Indeed, there
is a consensus regarding the importance of socioeconomic differences in explaining ill health
[3], and differences in education, employment, and income are consistently associated with
health inequalities [4–6]. In disadvantaged neighborhoods, the inadequate environmental and
infrastructural conditions, such as the presence of trash, open sewers, and standing water, also
contribute to the proliferation of pathogens and their hosts, and increase resident exposure to
a range of pathogens [7,8]. Funding: The study was funded by Medical
Research Council (UK) (MR/ P024084/1) to MB,
Fundac¸ão de Amparo à Pesquisa do Estado da
Bahia (BR) (1502/2008). The funders had no role in
the design, data collection, analysis, decision to
publish, or preparation of manuscript. In addition to contextual risk factors that directly or indirectly expose people to pathogens,
differences in knowledge, attitudes, and practices (KAP) towards diseases also contribute to
health disparities. Many recent studies highlight the importance of KAP for COVID-19 [9],
tuberculosis [10], malaria [11], leptospirosis [12], and arboviruses such as zika [13] and dengue
[14]. Most of these studies rely on the assumption that the risk of acquiring infection is deter-
mined by individual patterns of behavior (practices). These practices are assumed to be driven
by attitudes, which are determined, in turn, by the level of knowledge of a disease and its associ-
ated risk factors [10–14]. However, rarely has KAP been related to objective measures of risk
such as probabilities of infection computed from field data [15,16]. While a number of studies
have considered the role of demographic and socioeconomic factors in shaping KAP, the role of
environmental conditions has been largely overlooked. Furthermore, available KAP studies
have implicitly assumed that knowledge, attitudes, and practices all share the same determi-
nants. Indeed, there is an apparent absence of studies that have disentangled the direct effects of
socioeconomic [15,16], and environmental variables [17], on disease risk, and also their indirect
effects, acting separately, through knowledge, attitudes and practices in urban communities. Abstract Several studies have identified socioeconomic and environmental risk factors for infectious
disease, but the relationship between these and knowledge, attitudes, and practices (KAP),
and more importantly their web of effects on individual infection risk, have not previously
been evaluated. We conducted a cross-sectional KAP survey in an urban disadvantaged
community in Salvador, Brazil, leveraging on simultaneously collected fine-scale environ-
mental and epidemiological data on leptospirosis transmission. Residents’ knowledge influ-
enced their attitudes which influenced their practices. However, different KAP variables
were driven by different socioeconomic and environmental factors; and while improved KAP
variables reduced risk, there were additional effects of socioeconomic and environmental
factors on risk. For example, males and those of lower socioeconomic status were at greater
risk, but once we controlled for KAP, male gender and lower socioeconomic status them-
selves were not direct drivers of seropositivity. Employment was linked to better knowledge
and a less contaminated environment, and hence lower risk, but being employed was inde-
pendently associated with a higher, not lower risk of leptospirosis transmission, suggesting
travel to work as a high risk activity. Our results show how such complex webs of influence
can be disentangled. They indicate that public health messaging and interventions should
take into account this complexity and prioritize factors that limit exposure and support appro-
priate prevention practices. Editor: Syed Shahid Abbas, Institute of
Development Studies, UNITED KINGDOM Received: March 29, 2022
Accepted: November 7, 2022
Published: December 9, 2022 Peer Review History: PLOS recognizes the
benefits of transparency in the peer review
process; therefore, we enable the publication of
all of the content of peer review and author
responses alongside final, published articles. The
editorial history of this article is available here:
https://doi.org/10.1371/journal.pgph.0000408 Copyright: © 2022 Palma et al. This is an open
access article distributed under the terms of the
Creative Commons Attribution License, which
permits unrestricted use, distribution, and
reproduction in any medium, provided the original
author and source are credited. Data Availability Statement: The datasets used
and/or analyzed during the current study cannot be 1 / 14 PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 PLOS GLOBAL PUBLIC HEALTH Deconstructing leptospirosis transmission risk in Salvador, Brazil Introduction Strategies to reduce disease risk are often top-down and overlook local knowledge, and more
importantly, local barriers, facilitators of preventative measures, and risk perception [2]. Hence,
to identify appropriate intervention targets that are more consistent with the reality of the most
impoverished populations, it is crucial to trace and quantify the relationships between percep-
tions and practices of residents, measured through KAP, and the socioeconomic, demographic,
and environmental factors as both groups of factors combine, ultimately, to determine risk. Competing interests: The authors have declared
that no competing interests exist. In this study, we present an analysis aimed at disentangling these various and potentially
interrelated pathways to leptospirosis transmission risk in the city of Salvador, Brazil. Lepto-
spirosis is a globally distributed and neglected disease that disproportionately affects vulnera-
ble urban residents in low-income communities. Earlier, we have identified socioeconomic
and environmental risk factors for leptospirosis [7,18,19], and found a strong link between
socioeconomic status, infrastructure and sanitation on the one hand, and exposure to Leptos-
pira on the other. Here, we conducted a cross-sectional and detailed KAP study in an urban
low-income community, leveraging simultaneously collected fine-scale environmental and
epidemiological data, to understand how demographic, socioeconomic, and environmental
factors interact to determine KAP of residents, and how the combination of aforementioned
factors determines infection risk. Study area Our project, initiated in 2017, is based in four low-income communities in the city of Salvador,
Brazil (ca. 2.872.347 million inhabitants in 2019) [20]. In those four communities, we 2 / 14 PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 PLOS GLOBAL PUBLIC HEALTH Deconstructing leptospirosis transmission risk in Salvador, Brazil Fig 1. Socioeconomic and environmental profile of the study area, Marechal Rondon, Salvador, Bahia, Brazil. The
maps are not copyrighted as they were created by the authors using QGIS 2.18 software. The boundary polygons of
Brazil, Salvador and the production of the Salvador income map were downloaded from open and publicly accessible
base of IBGE—Instituto Brasileiro de Geografia e Estatistica, on the Geosciences platform which can be accessed at
https://www.ibge.gov.br/geociencias/downloads-geociencias.html. The WorldView-3 May 2017 satellite image with
31cm resolution was also used to digitize the study area and sewer line. The image was acquired by the research project
/ Instituto Gonc¸alo de Moniz—IGM—Fiocruz Bahia from the company Satmap, with disclosure permitted referencing
the Copyrights of DigitalGlobe images. https://doi.org/10.1371/journal.pgph.0000408.g001 Fig 1. Socioeconomic and environmental profile of the study area, Marechal Rondon, Salvador, Bahia, Brazil. The
maps are not copyrighted as they were created by the authors using QGIS 2.18 software. The boundary polygons of
Brazil, Salvador and the production of the Salvador income map were downloaded from open and publicly accessible
base of IBGE—Instituto Brasileiro de Geografia e Estatistica, on the Geosciences platform which can be accessed at
https://www.ibge.gov.br/geociencias/downloads-geociencias.html. The WorldView-3 May 2017 satellite image with
31cm resolution was also used to digitize the study area and sewer line. The image was acquired by the research project
/ Instituto Gonc¸alo de Moniz—IGM—Fiocruz Bahia from the company Satmap, with disclosure permitted referencing
the Copyrights of DigitalGlobe images. https://doi.org/10.1371/journal.pgph.0000408.g001 https://doi.org/10.1371/journal.pgph.0000408.g001 conducted a sero-survey of 1,318 residents in 2018, where trained phlebotomists and nurses
collected blood samples and applied individual and household-level questionnaires to docu-
ment sociodemographic and environmental (household and peri-domiciliary) factors [7]. In this study, we conducted an in-depth KAP survey in one of those four communities,
namely Marechal Rondon (MR), located in the northwestern periphery of Salvador (Fig 1). MR is considered a low-income urban community with poor social and environmental con-
ditions (infrastructure and sanitation), as well with high concentration of families in vulner-
able areas at risk [21]. Unadjusted seroprevalence for Leptospira in MR was 12.0% (n = 338)
[7]. PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 Data collection The items in our questionnaire were based on previous KAP studies on leptospirosis
[12,24,25], and adapted to the context and objectives of this study. The 67 items (S1 Appendix)
were organized into four main categories: (1) sources of information on leptospirosis, (2) vari-
ables related to knowledge (what leptospirosis is; transmission, signs and symptoms; complica-
tions; control and prevention practices), (3) attitudes (individual, household and peri-
domiciliary) and (4) practices (individual, household and peri-domiciliary). Demographic,
socioeconomic, and environmental data at household and peri-domiciliary level were available
from our previous study [7]. For the “knowledge” section, only residents who had heard of leptospirosis proceeded to
answer the rest of the questions; those who had never heard of leptospirosis were not consid-
ered in subsequent analyses. For answers given by participants to questions based on state-
ments that were true, each “correct” answer was assigned a score of 2, a “do not know” answer
was assigned 1, and an “incorrect” answer was assigned 0; for false statements the scales were
reversed [25]. For questions on attitudes and practices, we used a three-point Likert scale [25,26]. For
each question on satisfactory attitudes, a “yes” answer was given a score of 2, “maybe” a score
of 1, and “no” was given 0 score. For preventative (satisfactory) practices, a “yes” answer was
given a score of 2, a “sometimes” answer received a score of 1, and a “no” answer was given a
0. For questions on unsatisfactory attitudes and practices, the scales were reversed. We
summed the scores for each individual to obtain the total score for each dimension of KAP. Questions on attitudes were related, among others, to the perceived seriousness of the disease,
perceived vulnerability to infection, and importance of protecting oneself (for full list see S3
Table and S1 Appendix). For the practices dimension, the questionnaire sought information
on protective measures taken by residents to avoid contact with sources of environmental con-
tamination (e.g., wearing gloves when handling trash) and to reduce resources for rats within
or near the household (e.g., adequate trash disposal, food storage, etc.). The maximum score
for knowledge, attitudes, and practices sections was 60, 30, and 28, respectively. Study area First, we identified all residents that had participated in the serological survey (N = 338)
and, approached those eligible to participate in this KAP study (N = 304). We confirmed the
adequacy of sample size for this study based on the previously reported 12% seroprevalence for
leptospirosis with 95% CI and an assumed non-response rate of 10% [22,23]. Eligible residents
were men and women 15 years or more of age, who slept in the household for at least three
nights per week, and had participated in the sero-survey in 2018. Trained team members vis-
ited the residents between April and June 2019, explained the objectives of the research, before
inviting them to participate. Upon recruitment, the team applied a standardized questionnaire
to document KAP of the study participants giving the participants a sufficient time to recall. Participants <18 years of age provided written informed assent and their legal guardians pro-
vided written informed consent. 3 / 14 PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 PLOS GLOBAL PUBLIC HEALTH Deconstructing leptospirosis transmission risk in Salvador, Brazil Statistical analysis a Standard deviation (SD) (for means) and Interquartile ranges (for medians) are shown for continuous variables of
age and per capita household. a Standard deviation (SD) (for means) and Interquartile ranges (for medians) are shown for continuous variables of
age and per capita household. a Standard deviation (SD) (for means) and Interquartile ranges (for medians) are shown for continuous variables of
age and per capita household. b Defined by income-generating activities and without a formal contract. c Reported exposure to mud, refuse, flooding water or sewage water in the workplace. d Occupation as construction worker, refuse collector or mechanic, which is associated with a workplace
environment characterized by high rat infestation. e Data is missing for twenty-four respondents. https://doi.org/10.1371/journal.pgph.0000408.t001 b Defined by income-generating activities and without a formal contract. c Reported exposure to mud, refuse, flooding water or sewage water in the workplace. d Occupation as construction worker, refuse collector or mechanic, which is associated with a workplace
environment characterized by high rat infestation. e D
i
i i
f
f
d d Occupation as construction worker, refuse collector or mechanic, which is associated with a workplace
environment characterized by high rat infestation. e Data is missing for twenty-four respondents. Statistical analysis For all participants, scores for each dimension of KAP were analyzed as the number of points
obtained as a proportion of the maximum possible score [12,26]. We combined KAP data with
previously collected data on serostatus for leptospirosis, demographic, socioeconomic, and
environmental/exposure factors [7,20]. Here, demographic data consisted of the following var-
iables: “gender (male/female)”, “age (below/above 40)”, and “race (black or mixed/non-
black)”. Socioeconomic variables were “finished primary school (yes/no)”, “family income
(lower/higher than minimum wage)”, “employment status (employed/ unemployed”), and
whether a participant “knows someone who had leptospirosis (yes/no)”. Family income was
categorized based on whether it was above or below the minimum wage of R$ 950, as defined
by the Brazilian Institute of Geography and Statistics [20]. Data on race was also based on
IBGE data, which recognized five different races (white, black, mixed, indigenous, Asian), but
classified here as either “black” or “non-black” as the majority of residents were black or mixed
race (Table 1). The environmental/exposure variables were “Owns animals (yes/no)”, “Owns
backyard (yes/no)”, “Presence of trash within 10 meters of house (yes/no)”, “Presence of open
sewer within 10 meters of the house (yes/no)”, “Walked barefoot in the last 12 months (yes/
no)”, and “Had contact with sewage in the last 12 months (yes/no)” (See complete dataset
attached as S1 File). 4 / 14 PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 PLOS GLOBAL PUBLIC HEALTH Deconstructing leptospirosis transmission risk in Salvador, Brazil Table 1. Sociodemographic and environmental characteristics of study population in Marechal Rondon, Salva-
dor-Bahia, Brazil (n = 248). Frequency or Median
%, SD or IQRa
Characteristic
Demographics
Male gender
91
36.7
Age (in years) a
44.2
18.4
Socioeconomic indicators
Incomplete primary school
68
27.4
Black or mixed race
226
91.1
Household per capita income (in Brazilian Reais) a
300
77–485
Occupation (n = 86)
Informal work b
65
75.6
Work-related exposures (n = 86)
Contact with contaminated environment c
64
74.4
Risk occupation d
22
25.6
Individual exposure factors
Sighting of rat (last year > 10 meters of house) e
138
61.6
Walked barefoot (last year)
72
29.0
Contact with sewage water (last year)
72
29.0
Contact with mud (last year)
115
46.4 Table 1. Sociodemographic and environmental characteristics of study population in Marechal Rondon, Salva-
dor-Bahia, Brazil (n = 248). PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 Attitude and practices of study participants about leptospirosis Most of the participants viewed leptospirosis a serious disease (98.4%) and had worries about
being infected (95.2%). However, 16.9% were not worried about the presence of rats near their
household, and 19% responded that they would not participate in leptospirosis prevention and
control activities organized by local health centers (S2 Table). Participant responses suggested
generally satisfactory preventative practices; 91.9% answered that they avoid contact with
sources of contamination. However, 21.0% of the participants reported that they do not use
gloves or boots when being in contact with garbage or sewage water, and 49.6% used illegal
rodenticides to control rats in their homes (S3 Table). Knowledge of study participants about leptospirosis Most participants (92.2%) had previously heard about leptospirosis. Among those, 48.4% iden-
tified television/radio as the main sources of information, and 24.0% and 10.2% reported that
they obtained the information from neighbors and school, respectively (S1 Table). The major-
ity of participants who had heard about leptospirosis identified it as a disease associated with
rats (93.9%) and contact with rat urine, with walking barefoot (98.4%), contact with trash
(95.5%) and flood water (93.9%) as the main modes of transmission. However, 22.4%
answered that leptospirosis is transmitted via mosquito bites and 12.6% believed it can be
spread from person to person (S1 Table). Demographic, socioeconomic, and environmental characteristics of study
participants Out of 304 eligible residents, 248 (81.6%) participated in this study (2.3% had moved out of the
study site, 0.3% died, 4.6% refused to participate, and 11.2% were not found at home on three
or more separate visits). The majority of participants were female (n = 157, 63.3%), with a
mean age of 44.2 (range 15–102 years; see Table 1 for a summary of the demographic and envi-
ronmental variables). https://doi.org/10.1371/journal.pgph.0000408.t001 For each dimension of KAP and for seropositivity, we first fitted bivariate analyses using a
generalized linear model (GLM) with a binomially distributed error structure. KAP scores and
seropositivity were the response variables, and each of the demographic, socioeconomic, and
environmental factors were included as predictors. Predictors with p-value < 0.15 were then
included in the full model for each dimension of KAP. In addition to these variables, we
included knowledge score as a predictor for the attitudes model, and both knowledge and atti-
tudes scores as predictors for the practices model. Finally, in addition to three models (one for
each dimension of KAP), we fitted an infection model (GLM with a binomially distributed
error structure), with serostatus as the response variable and KAP as predictors, in addition to
demographic, socioeconomic, and environmental variables with p-value <0.15 in the bivariate
analysis for seropositivity. For all multivariate models, we used a mixed (forward and backward) selection approach
using the dredge() function in the statistical software R. To select the final model parsimoni-
ously in each case, we identified the model with the lowest value of Akaike Information Crite-
ria (AIC). If two or further models were within two AIC values of this minimum, we selected
the model with the fewest number of predictors. For the final GLM models, we tested for mul-
ticollinearity among the predictors using the variance inflation factor (VIF), where a VIF < 10
was considered acceptable [27]. We performed all analyses in R [28], using MuMin() and lme4
() packages [29,30] (See complete script attached as S2 File). 5 / 14 PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 PLOS GLOBAL PUBLIC HEALTH Deconstructing leptospirosis transmission risk in Salvador, Brazil Ethics statement This study was approved by the Research Ethics Committee of the Institute of Collective
Health/ Federal University of Bahia (CEP/ISC/UFBA) with CAAE number
68887417.9.0000.5030, and a National Research Ethics Committee (CONEP) linked to Brazil-
ian Ministry of the Health under approval numbers 2.245.914–2.245.914.17–3.315.568. Demographic, socioeconomic, and environmental drivers of the
knowledge, attitudes, and practices Knowledge score was associated with demographic, socioeconomic, and environmental fac-
tors. Knowledge scores were lower for males compared to females (OR = 0.83, 95% Cl = 0.76–
0.92), and for participants who did not finish primary education compared to those who did
(OR = 0.73, 95% Cl = 0.66–0.80) (Table 2). Knowledge scores were higher for participants who
were employed (OR = 1.13, 95%Cl = 1.02–1.24), who lived in households with a family
income > 950 R$ (OR = 1.14, 95% Cl = 1.04–1.24), and participants who personally knew
someone who had leptospirosis (OR = 1.23, 95%Cl = 1.12–1.35). Among the environmental
variables, having an unpaved backyard was associated with a lower knowledge score compared
to those who did not (OR = 0.86, 95% Cl = 0.79–0.95) (Table 2). 6 / 14 PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 PLOS GLOBAL PUBLIC HEALTH Deconstructing leptospirosis transmission risk in Salvador, Brazil Table 2. Final three multivariate binomial regression (GLM) models explaining knowledge score, attitudes score,
and practices score using demographic, socioeconomic, and environmental variables (n = 246). aOR (95% CI) a
P-value b
Variables
Knowledge Score
Male
0.83 (0.76–0.92)
<0.001
Did not finish primary education
0.73 (0.66–0.80)
<0.001
Employed: yes
1.13 (1.02–1.24)
0.018
Family income > R$ 950 (minimum wage)
1.14 (1.04–1.24)
0.006
Knows someone who had leptospirosis
1.23 (1.12–1.35)
<0.001
Has a backyard
0.86 (0.79–0.95)
0.002
Attitude Score
Knowledge score
1.03(1.02–1.05)
<0.001
Male
0.77 (0.64–0.91)
<0.001
Older than 40
1.36 (1.14–1.63)
0.018
Employed: yes
1.36 (1.13–1.63)
0.006
Did not finish primary education
0.80 (0.65–0.98)
<0.001
Race Black
1.72(1.35–2.18)
0.002
Owns animals
0.71 (0.60–0.84)
<0.001
Presence of trash within 10 meters of house
1.23 (1.05–1.45)
0.011
Practice Score
Attitude score
1.11 (1.09–1.13)
<0.001
Walked barefoot outside
0.81 (0.67–0.97)
0.020
Presence of trash within 10 meters of house
0.82 (0.69–0.98)
0.028
Contact with sewage in last 12 months
0.84 (0.70–1.01)
0.057
a aOR = Adjusted odds ratio. b p 0.05. https://doi.org/10.1371/journal.pgph.0000408.t002 Table 2. Final three multivariate binomial regression (GLM) models explaining knowledge score, attitudes score,
and practices score using demographic, socioeconomic, and environmental variables (n = 246). For attitudes, the final model included knowledge score as a predictor, whereby a higher
knowledge score was associated with higher, hence more satisfactory, attitudes score
(OR = 1.03, 95% CI = 1.02–1.05) (Table 2). PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 Demographic, socioeconomic, and environmental drivers of the
knowledge, attitudes, and practices Similar to the knowledge score model, demo-
graphic, socioeconomic, and environmental factors were also associated with attitudes score
beyond their indirect effects, acting through knowledge. Male participants were more likely to
have a lower attitudes score compared to female participants (OR = 0.77, 95% Cl = 0.64–0.91),
and those >40 years of age were more likely to have a higher attitudes score compared to
younger participants (OR = 1.36, 95% Cl = 1.14–1.63). Being employed was associated with a
higher attitude score (OR = 1.36, 95% Cl = 1.13–1.63), and participants who did not finish pri-
mary education had lower attitudes scores compared to those who did (OR = 0.80, 95%
CI = 0.65–0.98). Participants who consider themselves black race had higher attitudes score
compared with non-black participants (OR = 1.72, 95%Cl = 1.35–2.18). Among environmental
factors, ownership of animals was associated with a lower attitudes score (OR = 0.71, 95%
Cl = 0.60–0.84). On the other hand, the presence of trash within 10 meters of household was
associated with a higher attitudes score (OR = 1.23, 95% Cl = 1.05–1.45) (Table 2). For practices, the final model suggested that a higher practices score, and thus more satis-
factory practices, was associated with a higher attitudes score (OR = 1.11, 95% Cl = 1.09–1.13),
in addition to two environmental (but not demographic or socioeconomic) variables, which
had effects beyond their indirect effects, acting through knowledge and attitudes (Table 2). Lower practices scores were found for participants who had walked barefoot in the past 12 7 / 14 PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 PLOS GLOBAL PUBLIC HEALTH Deconstructing leptospirosis transmission risk in Salvador, Brazil months compared to those who did not (OR = 0.81, 95% Cl = 0.67–0.97) and who had contact
with sewage in the past 12 months (OR = 0.84, 95% Cl = 0.70–1.01), the latter association
being near significant. Participants who reported the presence of trash within 10 m of the
household also had lower practices scores than those who did not (OR = 0.82, 95% Cl = 0.69–
0.98) (Table 2). KAP as a driver for Leptospira seropositivity Among the 248 participants in this study, 29 (11.7%) were seropositive to leptospirosis. Partici-
pants’ >40 years of age were more likely to be seropositive for leptospirosis compared to youn-
ger participants (OR = 3.45, 95% Cl = 1.27–9.35), as were employed participants compared to
unemployed ones (OR = 2.45, 95% Cl = 1.03–5.86). Contact with sewage in the past 12 months
was also associated with a higher seropositivity (OR = 3.40, 95% CI = 1.43–8.07). Finally,
higher scores for knowledge and practices were associated with lower seropositivity
(OR = 0.89, 95% CI = 0.82–0.97 for knowledge; OR = 0.82, 95% CI = 0.70–0.96 for practices)
(Table 3). https://doi.org/10.1371/journal.pgph.0000408.t003 PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 Discussion In this cross-sectional study, we evaluated the contributions of knowledge, attitudes and prac-
tices (KAP), and their respective determinants, to Leptospira seropositivity in a disadvantaged
community in the city of Salvador, Brazil. Our final model for seropositivity indicated that res-
idents who had recent contact with sewage were three times more likely to be seropositive
compared to those who did not, and employed residents were more than twice as likely to be
positive compared to unemployed residents. However, seropositivity was also lower in resi-
dents with higher knowledge of the disease and for those who adopted protective practices. Crucially, knowledge and practices themselves were determined by a range of demographic,
socioeconomic and environmental factors. These direct and indirect pathways are summarized
in Fig 2. The scores for knowledge, affecting seropositivity indirectly, were lower in males, in those
who did not complete their primary education, and who had a back yard. Knowledge scores
were higher in those who were employed, had a family income higher than the minimum
wage, and who knew someone who had had leptospirosis. The scores for attitude were higher
in those with better knowledge, and so each of the factors affecting knowledge affected atti-
tudes indirectly. Attitude scores were also higher in residents over 40, those who were
employed, those who identified as black, and those with trash within 10 meters of their house,
and were lower in males, in those who failed to finish primary education, and those who Table 3. Final multivariate logistic regression (GLM) predicting seropositivity of residents using KAP, demo-
graphic, socioeconomic, and environmental variables (n = 246). aOR a (95% CI)
P-value b
Variables
Older than 40
3.45 (1.27–9.35)
0.015
Employment
2.45 (1.03–5.86)
0.044
Contact with sewage in last 12 months
3.40 (1.43–8.07)
0.006
Knowledge score
0.89 (0.82–0.97)
0.006
Practice score
0.82 (0.70–0.96)
0.016
a aOR = Adjusted odds ratio. b p 0.05. https://doi.org/10.1371/journal.pgph.0000408.t003 Table 3. Final multivariate logistic regression (GLM) predicting seropositivity of residents using KAP, demo-
graphic, socioeconomic, and environmental variables (n = 246). 8 / 14 PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 PLOS GLOBAL PUBLIC HEALTH Deconstructing leptospirosis transmission risk in Salvador, Brazil Fig 2. The network of relationships linking socioeconomic, demographic, and environmental factors to
knowledge, attitudes, practices scores, and showing how both groups of factors determine, directly and indirectly,
leptospirosis sero-positivity. Discussion The diagram represents the output from four generalized linear models, and the
individual variables can be found in Tables 2 and 3. The authors created these figures using the findings generated
from the analysis of their primary data. https://doi.org/10.1371/journal.pgph.0000408.g002 Fig 2. The network of relationships linking socioeconomic, demographic, and environmental factors to
knowledge, attitudes, practices scores, and showing how both groups of factors determine, directly and indirectly,
leptospirosis sero-positivity. The diagram represents the output from four generalized linear models, and the
individual variables can be found in Tables 2 and 3. The authors created these figures using the findings generated
from the analysis of their primary data. https://doi.org/10.1371/journal.pgph.0000408.g002 https://doi.org/10.1371/journal.pgph.0000408.g002 owned animals. Finally, the factors that drive practices affect exposure indirectly, and so each
of the factors affecting attitudes, including those acting indirectly through knowledge, affected
practices indirectly. Practices scores were also lower in those who walked barefoot, those with
trash within 10 meters of their house, and those who had come into contact with sewage in the
last 12 months. Of the many pathways and drivers of Leptospira seropositivity identified, a number of path-
ways are particularly notable. First, our earlier research has shown that males had a higher risk
of leptospirosis [7,18], a pattern commonly observed for several environmentally-transmitted
and vector-borne diseases [31–33]. Both behavioral and physiological differences between men
and women contribute to sex/gender biases in risk [34]. Also, cultural variables, such as gender
stereotypes, contribute to differentials in attitudes to risk [35]. Here, we found that men had
lower knowledge and attitudes scores compared to women, which contributed directly and
indirectly, through sub-optimal practices, to an observed higher risk of infection in men. How-
ever, once we controlled for KAP, male gender was not a significant (direct) driver of seroposi-
tivity, suggesting that gender-specific attitudes and exposure patterns [35], and not differences
in physiology, were responsible for the higher leptospirosis risk in males observed in previous
studies. Indeed, gender biases in leptospirosis risk become even larger in rural settings, due to
higher Leptospira exposure for men during agricultural and fishing activities [34]. In previous studies, lower scores for socioeconomic variables such as income and education
have been linked to a higher risk of leptospirosis and other zoonotic and vector-borne diseases
[7,18,19,36]. PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 Discussion Here too, higher family income, completing primary education, and being
employed were associated with higher knowledge scores, and being employed was also associ-
ated with better attitudes. Hence, higher socioeconomic status leads to better knowledge, atti-
tudes, and consequently more satisfactory practices that ultimately contributed to lower
seropositivity. These patterns are thus consistent with previous studies, indicating that even
after controlling for demographic and environmental factors, lower socioeconomic status was
a direct predictor of leptospirosis risk [7]. Notably, however, socioeconomic status here had no 9 / 14 PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 PLOS GLOBAL PUBLIC HEALTH Deconstructing leptospirosis transmission risk in Salvador, Brazil direct negative impact on seropositivity, suggesting KAP as the causal route to such health
inequalities. Furthermore, having employment was independently associated with a higher, not a lower
risk of Leptospira exposure (Fig 2). In an earlier study, we proposed that exposure away from
the household is understudied and remains difficult to account for [7]. Our present results
support this view, suggesting that while employment, among other proxies of higher socioeco-
nomic status, are linked to better knowledge and a less contaminated peri-domestic environ-
ment [7], work- or travel-related exposure away from the household remains inevitable given
the inadequate environmental and sanitary conditions that are ubiquitous in the community. Frequent large-scale flooding events that affect public and private spaces, inadequate infra-
structure, and poor access to main roads and public transit make travel to work and work itself
risky activities, with negative consequences on health and potentially income [37]. This inabil-
ity to avoid environmental contamination due to lack of alternatives is a common challenge in
low-income settings. In northern Senegal, for example, villagers were aware of schistosomiasis
risk in nearby lakes, but had no option but to continue fishing and agricultural activities in or
in close proximity to those lakes [38]. Residents who identified as black/mixed race had better attitudes towards leptospirosis
compared to non-black residents. However, there were no differences between the two groups
in knowledge, practices, or seropositivity, in contrast to previous studies, which showed that
black/mixed residents were at a higher risk of leptospirosis [18]. Our findings therefore suggest
that while residents of black/mixed ethnicity disproportionately recognized leptospirosis as a
potentially serious health issue, they were at a similar (and in previous studies higher) risk of
leptospirosis compared to non-black residents. PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 Discussion Our findings are in line with those reported for
other diseases and contexts. Historically marginalized ethnic groups in the US were more
likely to perceive COVID-19 as a major threat to society [39], yet incidence and severe
COVID-19 infections are disproportionately higher in marginalized ethnic minorities [40]. Hospitalizations due to infectious diseases in general are higher for indigenous ethnic commu-
nities, especially for the most deprived groups [41]. Here, despite the better attitudes observed
towards disease risk, socioeconomic and environmental inequalities, manifested by types of
employment, living and environmental conditions, and sociodemographic factors, kept mar-
ginalized ethnic groups at a disproportionate risk of infectious diseases [42]. Previous studies on leptospirosis have reported that poor environmental and sanitary con-
ditions increase the risk of infection [18,19,24,43]. Here we found an additional, indirect con-
tribution of poor environmental conditions to Leptospira risk, acting through KAP. For
example, while the presence of trash near the household contributed to better attitudes towards
leptospirosis, as it likely made residents aware of high levels of rat infestation, it nevertheless
made residents less likely to adopt satisfactory practices. Further, recent exposure to sewage, in
addition to directly contributing to higher seropositivity, also made residents less likely to
adopt satisfactory practices. Indeed, no socioeconomic or demographic factor directly influ-
enced practices, and thus their effect was strictly indirect, mediated through knowledge and
attitudes scores. Inadequate environmental conditions, therefore, appeared to form barriers
against the adoption of protective practices. We hypothesize that a degraded peri-domiciliary
environment, e.g., the presence of trash piles near the household, despite being associated with
better attitudes, results in protective practices being seen as useless or of limited value given
the overwhelming physical reality that expose residents to a wide range of risk factors. In
future studies, residents should be specifically asked about their degree of belief in the effec-
tiveness of protective measures, which could then be tested against environmental and infra-
structural conditions near the household. Indeed, variables representing inadequate
environmental conditions were doubly important, as they not only contributed to PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 10 / 14 PLOS GLOBAL PUBLIC HEALTH Deconstructing leptospirosis transmission risk in Salvador, Brazil seropositivity indirectly by making residents less likely to adopt preventative practices, but also
directly, after KAP had been taken into account. This study has several limitations. Conclusion Our results indicate that to understand the links between KAP and disease risk, we need to
account for the pathways by which demographic, socioeconomic, and environmental factors
influence the KAP dimensions and the risk of human leptospirosis, and the options available
for residents to mitigate it. Irrespective of factors such as education, income, knowledge, atti-
tudes, and protective measures taken by residents, the inadequacy of the environment and
socioeconomic disadvantage remain central, as they render residents helpless in avoiding
exposure and ultimately becoming infected. Our findings are important for identifying poten-
tial barriers that limits prevention practices and continue to expose residents, and suggest that
public health messaging and interventions should take into account both the socioeconomic
and environmental determinants of at-risk groups. S1 Table. Summary of knowledge questions on leptospirosis (n = 246).
(DOCX) S2 Table. Summary of attitudes questions on leptospirosis (n = 248). (DOCX) S3 Table. Summary of practices questions on leptospirosis (n = 248). (DOCX) S3 Table. Summary of practices questions on leptospirosis (n = 248). (DOCX) Discussion The first is the impossibility of establishing causality for
the associations presented here given the cross-sectional design of the study. Another limita-
tion is related to memory bias of participants, since the variables related to exposure in the
year were self-reported. Acknowledgments We deeply appreciate all the residents and community leaders of the neighborhood Marechal
Rondon for their contributions in supporting the research and participation in this study. We
thank the members of the project for their constructive suggestions for this manuscript. We deeply appreciate all the residents and community leaders of the neighborhood Marechal
Rondon for their contributions in supporting the research and participation in this study. We
thank the members of the project for their constructive suggestions for this manuscript. S1 Appendix. List of KAP questions.
(DOCX) S1 Appendix. List of KAP questions. (DOCX) Supporting information Supporting information
S1 Checklist. STROBE statement—checklist of items that should be included in reports of
cross-sectional studies. (DOC)
S1 Appendix. List of KAP questions. (DOCX)
S1 Table. Summary of knowledge questions on leptospirosis (n = 246). (DOCX)
S2 Table. Summary of attitudes questions on leptospirosis (n = 248). (DOCX)
S3 Table. Summary of practices questions on leptospirosis (n = 248). (DOCX)
S1 File. Complete dataset. (R)
S2 File. Complete script. (TXT) S1 Checklist. STROBE statement—checklist of items that should be included in reports of
cross-sectional studies.
(DOC) S1 Checklist. STROBE statement—checklist of items that should be included in reports of
cross-sectional studies. S1 Checklist. STROBE statement—checklist of items that should be included in reports of
cross-sectional studies. (DOC) Author Contributions Conceptualization: Fabiana Almerinda G. Palma, Federico Costa, Michael Begon, Hussein
Khalil. 11 / 14 PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
December 9, 2022 PLOS GLOBAL PUBLIC HEALTH Deconstructing leptospirosis transmission risk in Salvador, Brazil Data curation: Fabiana Almerinda G. Palma, Federico Costa, Daiana Santos de Oliveira, Hus-
sein Khalil. Formal analysis: Fabiana Almerinda G. Palma, Federico Costa, Fa´bio Neves Souza, Hussein
Khalil. Funding acquisition: Federico Costa, Mitermayer G. Reis, Albert I. Ko, Michael Begon. Investigation: Fabiana Almerinda G. Palma, Federico Costa, Daiana Santos de Oliveira, Hus-
sein Khalil. Methodology: Fabiana Almerinda G. Palma, Federico Costa, Hussein Khalil. Project administration: Fabiana Almerinda G. Palma, Federico Costa, Mitermayer G. Reis,
Albert I. Ko, Hussein Khalil. Resources: Fabiana Almerinda G. Palma, Federico Costa, Ricardo Lustosa, Daiana Santos de
Oliveira, Hussein Khalil. Software: Fa´bio Neves Souza, Hussein Khalil. Supervision: Federico Costa, Mitermayer G. Reis, Albert I. Ko, Michael Begon, Hussein
Khalil. Validation: Federico Costa, Michael Begon, Hussein Khalil. Visualization: Fabiana Almerinda G. Palma, Ricardo Lustosa, Fa´bio Neves Souza, Hussein
Khalil. Writing – original draft: Fabiana Almerinda G. Palma, Federico Costa, Hammed O. Mogaji,
Fa´bio Neves Souza, Michael Begon, Hussein Khalil. Writing – review & editing: Fabiana Almerinda G. Palma, Federico Costa, Hammed O. Mogaji, Fa´bio Neves Souza, Michael Begon, Hussein Khalil. PLOS Global Public Health | https://doi.org/10.1371/journal.pgph.0000408
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https://openalex.org/W2017701290
|
OpenAlex
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Open Science
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Public Domain
| 1,912
|
THE TREATMENT OF WOUNDS, WITH REFERENCE TO TETANUS PROPHYLAXIS
|
Oscar Berghausen
|
English
|
Spoken
| 2,252
| 3,994
|
SERUM
REACTIONS
l g
2. Cleanse the surrounding parts with green soap, alcohol,
ether and sterile water. These were noted
in several
eases and
were marked
by local redness, swelling, urticaria-like eruptions and
fever. Owing to patients not reporting
as directed,
we
were unable to obtain complete statistics
in
this
regard. With the aim of preventing such symptoms or
of ameliorating
them
when
once developed, air-pin
sulphate (gr. 1/100-1/120 three times
a day. in chil-
dren less) was given hypodernnitically, particularly when
numerous injections of
serum
were made
in
eases of
developed
tetanus. We
have
found
thai
ihis
drug
possesses undoubted value in preventing such symptoms. Itching, redness and urticarial eruptions frequently dis-
appeared when atropin was given. We therefore adopted
this measure
as
¡t routine before all repeated injections
of scrum. At times such eruptions will appear follow-
ing the
use of atropin sulphate, hut never lo
a
very
marked degree. It. Remove with sterile fórceps any foreign material lying
superficially in the wound. p
y
4. Cleanse
the wound
with
5 per
cent, phenol
(carbolic
acid)-0.5 per cent, hydrochloric acid solution. i )
p
y
;">. Enlarge
the opening by
free
incision
if
necessary
to
thoroughly cleanse the wound, or for the removal of foreign
substance. (i. Use
a
general
anesthetic whenever
indicated. P
k g
7. Pack
the
wound
lightly
with
gauze
soaked
in
the
phenol-hydrochloric acid solution, and dress, (.'hange the dress-
ings daily. g
y
8. Immediately after dressing fhe wound
on the first day
give 1,500 units of antitetanie
serum subcutaneously. This
serum
can be obtained at the laboratory. l b y
9. A careful record must be kept and sent to the labora-
tory when the patient is discharged. y
p
g
10. In tlie case of doubt or on the appearance of symptoms
resembling tetanus, notify
nie
| Ucrghnusen] at once. TABLE 1.—CASES IX WIIKTI BEEUM WAS USED PROPHXLAC-
TICALLY; GOOD RESULTS
Total
No. Serum Used,
Character of Injury
of fuses
in TInlls
I'linclureil wounds (mostly ninile li.v iinllsl
. . . . 72
león
coniused und lacerated.'.. 4
1500
( 'iinnon-cracker wounds . 2
1600
(lun-sbot wounds. u
ir.nu
Blank-cartridge wounds . 7
1800
1 'owder burns. 5
1500
no TABLE 1.—CASES IX WIIKTI BEEUM WAS USED PROPHXLAC-
TICALLY; GOOD RESULTS Heretofore such injuries
were opened, cleaned
and
treated with strong phenol solution. AN TIT HTA NIC KK1ÎUM
AS
A
I'DOl'l I VLACÏIC MEASURE Owing to the experience of others, we have used only
one injection of 1.500 units, usually given in that part
of the anatomy nearest the wound. We repeated the
injection in three eases in which suppuration persisted. In this connection
we wish lo quote the results of Sir
D. Semplc,1 who says: OSCAR BERGHAUSEN, B.A., M.D. AND
CHARLES E. HOWARD, M.D. Receiving Physician at the Cincinnati Hospital
CINCINNATI In June, 1910, the late Dr. N. P. Dandridge
proposed
the systematic handling of all wounds, punctured, pene-
trating or lacerated, with the aim of ascertaining the
best methods of treating such
cases at
a large general
hospital, particularly with reference to tetanus prophy-
laxis. As is well known, to avoid
the development of
tetanus we must begin by treating, in
a thorough sur-
gical manner, the wound received. The
use of anti-
tetanic serum
as
a prophylactic agent
was resorted to
in
a large series of cases, in order to test its value. Th
f i
i
i Many people in apparently good health
harbor
Bpores of
tetanus in healed wounds or in the intestinal tract, and that
hidden away in the tissues the spores remain alive and retain
their virulence, hut
do not. grow into toxin-producing bacilli-
Th
h ,
g
p
g
. . . The leukocytes
do
not always destroy
(lie spores,
hut
when
a local suppuration
has ceased they may
be
able
to
wander away with
the spores still
in
them. . . . Spore-car-
riers
arc in danger of suffering from
tetanus
(a)
on the
occurrence of great fatigue or exposure to heat or cold, which
diminish their resistance; (h) when the site where Ihe spores
are lodged
becomes
converted
into
a medium,
which
from
being anaerobic and from
a failure of phagocytosis, is favor-
able for the growth of the spores into toxin-producing bacilli:
and (c) when a focus of dead tissue forms in
a part of the
body at a distance from the site where Ihe spores are lodged, a ge
The following list of instructions were placed in each
surgical ward : The interns will pleasecarry
out the instructions mentioned
below for the following classes of
cases. f
i g
1. All perforating, penetrating
or lacerating wounds
con-
taminated
directly by
soil
or manure, especially those
con-
tracted in the streets or about stables. 1. Semple,
Sir D.
:
The Relation of Tetanus to the Hypodermic
or Intramuscular Injection
of Quinin,
Paludism, Simla, January
1911.
No. 2, p. 32. AN TIT HTA NIC KK1ÎUM
AS
A
I'DOl'l I VLACÏIC MEASURE Quinin given hypodermatic-ally
may produce
a local
tissue necrosis; soluble non-irritating substances do not. Semple further asserts that from 10 to 15 c.c. of anti-
tëtanic serum renders a patient passively immune for
R
period of from two lo three weeks, and has found if
;'
valuable prophylactic agent
when using quinin liypo-
dermaticálly. 2. All blank-cartridge
and giant-cracker perforating
and
lacerating wounds. Downloaded From: http://jama.jamanetwork.com/ by a University of Iowa User on 05/24/2015 INSTRUCTIONS 1. In all
cases above mentioned
remove the clothing and
foreign material about the wound. these were found to be cathei loosely made and could
easily be torn into fragments. From the Serological Department, Cincinnati Hospital. THE
TREATMENT
OF
WOUNDS,
WITH
REFERENCE
TO
TETANUS
PROPHYLAXIS
OSCAR BERGHAUSEN, B.A., M.D.
AND
CHARLES E. HOWARD, M.D.
Receiving Physician at the Cincinnati Hospital
CINCINNATI these were found to be cathei loosely made and could
easily be torn into fragments. TADLE
2.—CASUS
IN
WHICH
SERUM
WAS
NOT USED
I'liOlMIYLACTICALlA;
GOOD
RESULTS TADLE
2.—CASUS
IN
WHICH
SERUM
WAS
NOT USED
I'liOlMIYLACTICALlA;
GOOD
RESULTS
Character ol'Injury
No. Cases
Result
Punctured wound—nail . 3
Good
liliiiik-cartriilge . .'1
Good
0 Character ol'Injury
No. Cases
Result
Punctured wound—nail . 3
Good
liliiiik-cartriilge . .'1
Good
0 appeared
If antitetanio serum is to be used at. all. large doses
must be employed, although enormous doses,
as
recom-
mended by those interested in its sale,
were not used
by us. by
Atropin sulphate has some value in controlling senmi
reactions and is deserving of further trial. Th In Table 3 will be found those cases in which serum
was not used as a prophylactic measure, and which later
developed tetanus. g
The
subcutaneous administration
of phenol,
2
per
cent, solution, is followed by
an early appearance of
albumin in the urine. This possibility of damaging Ihe
kidneys must be taken into consideration when the injec-
tions
arc used. In
the future
we shall follow
such
injections by the rectal administration of
a liypertonio
neutral salt solution to limit, if possible, this damage
to flu; kidneys, in accordance with the results obtained
in experimental nephritis, by Prof. Martin II. Fischer,
of this city. de e oped
These patients were first seen by us after tetanus had
developed, excepting
eases
1 and 6. Case
7 occurred
in
the private practice
of
Dr. George Krieger
of
Madisonvillc, by whom we were consulted and to whom
We are indebted. Case 5 occurred in the service of Dr. Casper Hegner at the City Hospital. p
g
y
p
Two cases in which scrum was used prophylactically,
the one caused by
a cannon-cracker wound of the neck
and followed by extreme celliilitis before admission, the TAREE 8.—CASES
IN Willen
SUItUM
WAS
NOT USED
Htoi'll YEACTICAI.I.Y
AND
TETANUS
DEVELOPED
Cases
Character of injury
a
a3
'1. ¿?i
Sis
.2 SS
•sis
3 °5 D
3 » o °
£&5 E
— in
si
Ob
g§«
Mnltliile puncture wonnil ninile hy shot,,
. . 7
0
Lacerated wound . -i
—
20,000
Punctured wound (nail). n
—
0
Punctured wound (plck-ux). 8
,'1,000
Cut by a barbed wire. M
BO.OOO
Crushed fool:. 10
4-
27,000
Cut by a scylhe. 8
0
27,000
Compound fracture; infected. 0
—
27,000
0
0.24
0.14
(I
II
II
0
0
II hours
5 days
. .10 hours
4 days
,
14 days
. 2 days
. TADLE
2.—CASUS
IN
WHICH
SERUM
WAS
NOT USED
I'liOlMIYLACTICALlA;
GOOD
RESULTS About 2
1
2 days
. Death. Death. 'Death. Death. Recovery. Death. Recovery,
Death. * la (his column
-|
"doubtful."
means "good,"
— means "pool1" und 0 moans "none."
t 'n this column -|-
means "present,"
— "absent" and Willen
SUItUM
WAS
NOT USED
Htoi'll YEACTICAI.I.Y
AND
TETANUS
DEVELOPED other, caused by stepping
on
a nail and followed by
cellulitis before admission, resulted in sudden death. No autopsy could be secured, but death
was evidently
due to an embolus, no symptoms of tetanus developing
and
no anaphylactic phenomena. other, caused by stepping
on
a nail and followed by
cellulitis before admission, resulted in sudden death. No autopsy could be secured, but death
was evidently
due to an embolus, no symptoms of tetanus developing
and
no anaphylactic phenomena. We feel, after
a careful study of such injuries
as
listed in the foregoing, that no wound of such a nature
should bo treated lightly by any physician. By
care-
fully cleansing each wound, using a general anesthetic
if necessary to remove all foreign material, and employ-
ing
a diluted antiseptic to prevent sepsis, and
then
treating each one as an open wound, the physician has
done much
lo prevent
tetanus. By employing
one
immunizing dose of 1,500 units of antitctanic serum,
to be repeated only when suppuration has not ceased
after a week, he can practically assure the patient per-
fect safety from tetanus, Downloaded From: http://jama.jamanetwork.com/ by a University of Iowa User on 05/24/2015 injections (2 per cent.) included. Briefly slated,
our
observations are as follows: injections (2 per cent.) included. Briefly slated,
our
observations are as follows: In Table 2 will be found those cases in which serum
was not used
as
a prophylactic measure, and in which
the local treatment of the wound
was good. Of this
list not one developed tetanus. Although the
cases with
a short incubation period
offer the least hope, such cases are not necessarily fatal. Case
7
was
seen by Dr. Krieger, the first physician
consulted in the case, two clays after symptoms of tetanus
had developed,
and yet the manifest symptoms
dis-
appeared after one week's careful treatment. SERUM
REACTIONS
l The object of using
the hydrochloric-phenol mixture as recommended in the
text-books was to test ils efficiency, since it is attended
by less necrosis of tissue. Experience has shown
us
that treatment of all wounds after the above fashion
was sufficient. Particular care was taken to clean ovit all
wounds thoroughly, opening by incision if necessary, to
remove all foreign material. Particularly is this neces-
sary in blank-cartridge wounds when wads may have
entered. In
such
cases, in two instances, wads
were
removed
on successive days by different interns, each
one thinking that he had removed the last wad. In
llicse cases a general anesthetic, may become necessary. i
h
d
i
h In Table 1 will be found a list of cases of patients
treated at the hospital
with
the aim
of preventing
tetanus after the method described in the foregoing. N
f
d
l
d foregoing. Not one case of tetanus developed in the above series
of patients. ge e a
y
y
To ascertain
how
many wads
were present
in
each
blank carl ridge, four different specimens
were bought
and examined. In each one two wads were found, hut Results
with
and
without
prophylactic
serum
treatment In the ninety-six cases properly treated locally and by
the prophylactic administration of antitctanic
serum,
not one patient developed tetanus. patient developed
In the fourteen cases (Tables 2 and 3) treated with-
out the prophylactic administration of antitctanic serum,
fight patients developed tetanus of whom six died. h
) y
10 West Seventh Street. g
pat e ts de e oped
In the
cases (only six, however) properly treated,
locally and without the prophylactic administration of
antitctanic serum, not
one patient developed tetanus. P i g
p
p
In the
cases (only six, however) properly treated,
locally and without the prophylactic administration of
antitctanic serum, not
one patient developed tetanus. Patient G (Table 3)
was thoroughly treated locally,
out did not receive the prophylactic serum injection and
succumbed to tetanus. Public Health.--Public health is the foundation
on which
rests the happiness of the people anil the power of the state. Take the
most beautiful kingdom, give it intelligent
and
laborious citizens, prosperous manufacturers, productive agri-
culture; let arts flourish, let architects cover the land with
temples and palaces; in order to defend all these riches, have
iirst-rato weapons, fleets or torpedo boats
— if the population
remains stationary, if it decreases yearly in vigor and stature,
the nation must perish. And that is why I consider that the
first duty of
a statesman
is the
care
of public health.—
Disraeli. ,
patient developed
Patient G (Table 3)
was thoroughly treated locally,
out did not receive the prophylactic serum injection and
succumbed to tetanus. We had the opportunity of assisting in the treatment
Of eight patients with developed tetanus during the past
fifteen months, and feel that the information gained
l8 of some value. The usual methods of treatment were
employed, antitetanic
serum and phenol subcutaneous
| 45,565
|
https://openalex.org/W1971863697
|
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|
Open Science
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| 1,905
|
The Rhætic and Contiguous Deposits of Glamorganshire
|
L. Richardson
|
English
|
Spoken
| 26,042
| 54,649
|
1 Quart. Journ. Geol. Soc. vol. lxi (1905) pp. 374-84.
Q.J.G.S. No. 243.
2 r 385 385 By LI~SDALL RICHARDSOn, F.G.S. (Read May 24th, 1905.)
[PLATE XXXIII--FossIL~.]
CONTENTS,
Page
I. Introduction ............................................................ 385
II. The Sully Beds ........................................................ 386
III. Description of Sections. i. The Penarth District .......................................... 388
(A) Lavernock. (B) St. Mary's-Well Bay, Sully. (C) Cross Farm, near Dinas Powis. ii. Barry to Cowbridge .......................................... 397
(A) Coldknap, Barry. (B) Cadoxton. (C) Tregyff, near Cowbridge. iii. Cowbridge to Pyle ............................................. 406
(A) ~lendre, near Pencoed. (B) Quare]la Quarry, Bridgend. (C) Stormy Down. (D) Stormy-Down Cement-Works. IV. Additional Observations on the Sully Beds ..................... 410
V. Conditions of Deposition of the Rh-etic Black Shales ...... 414
VI. On the White Lias of Glamorganshire, and on the Strati-
graphical Position of the White Lias ........................ 415
VII. Summary. .............................................................. 420
VIII. Pal~eontological Notes
............................................. 422 Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from ~r,. L. P, IClqARDSO.N OH THE RH27ETIC AND
FAng. x9o5, Gloucestershirc and Worcestershire of certain beds at the base of
the Rhmtic Series. I then stated that ' there is evidence to suggest that there were earth-pressures at work at the close
of the Keuper Epoch, which caused the deposits to be thrown into slight ~vnelinal
and anticlinal flexures. In the depressed [n.eas the earlier'deposits of the Rhmtic
were laid down, and successive overlap on to the marls seems to have taken
place.' 1 My reasons for suggesting this explanation for the phenomenon
wero as follows. Throughout :North-West Gloucestershire and
Worcestershire the component deposits of the Rh~etic are remarkably
persistent. There were slight earth-movements towards the close of
the Pteria (Avic~daj-contorta age, it is true, which affected the persis-
tency of one or two beds ; but the cause for their irregularity is so
obvious, that it does not influence the matter under discussion. Once below a certain stratum, however, such persistency is not to
be observed. That stratum happens to be full of saurian-and
fish-remains at certain localities, on which account it has been
denominated the 'Bone-Bed.'
Therefore I employed this term,
or that of 'Bone-Bed equivalent,' according ms the circumstances
demanded. The geographical distribution o[ these infra-Bone-Bed
deposits appears to me to be, at present, satisfactorily explained
only by the theory that I have formulated. y
y
y
]:'or some years previous to the publication of the above-mentioned
paper, I had come to the conclusion that ' when the Rh~etic ocean gained access to the British area it spread over an
undulating expanse of Keuper Marls. In some areas, however,.., lakes
probably existed, and it would be in these areas that the complete sequence
from the :Keuper to lRhmtic deposits should be looked for. The section of
deposits formed under the conditions stated would be essentially of transitional
nature, as at Watchet; but where the Rha~tic ocean spread over the sur-
rounding ground a non-sequence would result. Thus, at the present time, the
junction-line would appear sharply defined; there would be no transitional
signs, and practically no erosion.' 2 A stretch of waier, probably of considerable dimensions, extended
into the Lavernock district, and therefore, according to my
theory, transition-beds
should be present. Such is the
case: the transition-beds are the 'Grey ~Iarls'(laars ) of the late
Robert Etheridge. This term would have been a convenient
designation for the beds, had it not been generally misinterpreted. ~r,. L. P, IClqARDSO.N OH THE RH27ETIC AND
FAng. x9o5, But, as such is the case, it appears desir;tble to replace it by the
term 'Sully Beds,' after the locality where the most interesting
development can be studied. 1 Quart. Journ. Geol. Soc. vo). lx (1,004) p. 356,
"- Ibid. p. 357. I. INTRODUCTION. IN the autumn of 1904 I made, in company with my friend,
Mr. E. Talbot, Paris, a detailed examination of the Rhmtic and
contiguous deposits of Monmouthshire and Glamorganshire. The
results of my investigations in the former count)" have already
been made known, ~ and it now remains to record those in the
latter. Less than four years have elapsed since the officers of the
Geological Survey completed the re-survey of the Secondary rocks of
Glamorganshire ; and that part of the memoir on 'The South-Wales
Coalfield ' which deals with the Bridgend district was only issued to
the public last January. This being the case, it might be thought
that but little of interest would have remained to be noticed in
the present communication. However, whereas the officers of the
Survey were mainly concerned in the mapping of the deposits,
my attention was directed ill most cases to the accumulation of
facts bearing upon the physical geography of the Rh~etic Epoch. g
p
p y
g
g
p y
p
On a previous
occasion I
communicated to this Society a
theory, to account for the geographical distribution in North-West 2 r Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from 386 ~r,. L. P, IClqARDSO.N OH THE RH27ETIC AND
FAng. x9o5, II. ThE SULT.Y BEDS. In his very interesting memoir of Robert F, theridge, Mr. H. B. Woodward, "F.]{.S., drew attention to the fact that the 'Grey
3Iarls ' and ' Tea-Green Marls' of Etheridge were distinct deposits. Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from 387 Vol. 6I.] co~T~vovs DEPOSITS OF OLA:MOR6AI~'SHIRE. ' Etheridge,' he wrote, ' originally placed them [that is, the ' Tea-
Green Marls'] in the Keuper, and distinguished them from the
Grey Marls which frequently form the base of the Rhmtic.'~
In
support of this interpretation of Etheridge's remarks, Mr. Wood-
ward quoted from the paper published by Etheridge in the
Proceedings of the Cotteswold Naturalists' Field-Club. In that
paper, Etheridge recorded this sequence:--' alternating bands of
grey and red fissile and conchoidal marls (No. 1 in section),
apparently here containing no fossils' ;2 and then, above these
'Tea-Green
Marls' a series of Grey Marls. The point upon
which Mr. Woodward naturally laid stress was that Etheridge
denominated the
lower of these two series the ' Tea-Green
Marls': not the upper, which he distinguished by the name
of 'Grey Marls.'
As will be seen by referring to his seetio~
at Garden Cliff, the beds that he wished to be called the' Tea-
Green Marls' are not those to which the term has been restricted
by most authors,
Etheridge's suggestion with regard to these
' Tea-Green Marls' of his at Garden Cliff was, that although
they did differ lithically, they nevertheless corresponded
to
certain beds at Watchet, Penarth, and Puriton, ' Etheridge,' he wrote, ' originally placed them [that is, the ' Tea-
Green Marls'] in the Keuper, and distinguished them from the
Grey Marls which frequently form the base of the Rhmtic.'~
In
support of this interpretation of Etheridge's remarks, Mr. Wood-
ward quoted from the paper published by Etheridge in the
Proceedings of the Cotteswold Naturalists' Field-Club. In that
paper, Etheridge recorded this sequence:--' alternating bands of
grey and red fissile and conchoidal marls (No. 1 in section),
apparently here containing no fossils' ;2 and then, above these
'Tea-Green
Marls' a series of Grey Marls. The point upon
which Mr. 2 Vol. iii (1865) pp. 220, 221. (
)pp
9
~ Proc. Cottesw. Nat. F.-C. vol. xiv 1 Proe. Bristol Nat. Soe. ser. 3, vol. x (1903-04) p. 183.
2 Vol. iii (1865) pp. 220, 221.
9
~ Proc. Cottesw. Nat. F.-C. vol. xiv (1903) table iii, facing p. 174. (
)pp
,
9
~ Proc. Cottesw. Nat. F.-C. vol. xiv (1903) table iii, facing p. 1 H. T. De la Beche, Mere. Geol. Surv. vol. i (1846)p. 253; T. Wright,
Quart. Journ. Geol. Soc. vol. xvi (1860) pp. 381-82; W. V. Guise (Presi-
dential Address, 1863), Prec. Cotteswold Nat. F.-C. vol. iii (18(15) pp. 117, 118 ;
H. W. Bristow, Geol. Mag. vol. i (1864) p. 236, & Rep. Brit. Assoc. 1.864
(Bath) Trans. Sections, p. 50; R. Etheridge, Trans. Cardiff :Nat. See. re/. iii
(1870-71) pt. ii, p. 39; Bcistow & Etheridge, Geol. Surv. Vertical Sections,
1873, Sheet 47 ; T. "Wright, ' Monogr. Lias Ammon. Brit. Is.' Pal. Soc. 1878,
p. 10; J. Storrie, Trans. Cardiff ~'at. Soc. vol. xiv (1882-83)p. 100; H. B.
Woodward, PL'oc. Geol. Assoc. vol. x (1888) p. 529, & Rep. Brit. Assoc. 1888
(Bath) p. 900; J. Storrie, Trans. Cardiff Nat. Soc. vo]. xxvi (1894)p. 105;
A. Strahan & T. C. Cantrilt, ' The Geology of the South-~Vales Coal/]eld:
Part ill--The Country around Cardiff' Mere. Geol. Surv. 1902, pp. 59-63. II. ThE SULT.Y BEDS. Woodward naturally laid stress was that Etheridge
denominated the
lower of these two series the ' Tea-Green
Marls': not the upper, which he distinguished by the name
of 'Grey Marls.'
As will be seen by referring to his seetio~
at Garden Cliff, the beds that he wished to be called the' Tea-
Green Marls' are not those to which the term has been restricted
by most authors,
Etheridge's suggestion with regard to these
' Tea-Green Marls' of his at Garden Cliff was, that although
they did differ lithically, they nevertheless corresponded
to
certain beds at Watchet, Penarth, and Puriton, ' at which places,' he wrote, ' I have termed them " Tea-Green Marls," from
the peculiar hue of the freshly-fractured shales when exposed, and the con-
stancy of their conditions.' The 'Grey Marls' (the upper strata of which are here designated
the Sully Beds) above he regarded as belonging to the Rheetic,
because the y contained fish-remains. y
Mr. Woodward's explanation of Etheridge's conclusions is very
satisfactory, because I had been compelled to adopt the view that
there were marls which were ]{h~etic, and again such as were
Keuper. But, while agreeing with Etheridge that this was the case,
I fail to see any evidence, either palmontologieal or stratigraphical,
why the marls below what I have called the ' Tea-Green Marls' at
Garden Cliff (which there come immediately below the Rh~etic
Black Shales)3 should be regarded as the equivalent of certain beds
which Etheridge admitted were lithically different at Penarth,
Watcher, and Puriton. Is it not much more probable that the' Grey
Marls '--certainly that the fossiliferous portion of them, namely,
the Sully Beds--of the Lavernock and Watchet districts--are not
represented in North-West Gloucestershire and Worcestershire;
and that they are only found within the limits of those
areas which
were
submerged
at the time
when the
Rhmtic ocean gained
access to the British
low-lying
country? y
These Sully Beds will be again dealt with after the sections in
Glamorganshire have been described. 2F2 Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from 388 (A) Lavernock. The section at Lavernoek is certainly one of the finest in the
country of the beds under consideration. The Rh,'etic can be
studied in the foreshore and cliffs of the deeply-indented little bay
immediately to the north of Lavernock Point; the White Lias
and the Ostrea-Beds succeed ; .while round the Point, and as far as
St. Mary's-Well Bay, Sully, are the pla~o~.bis-, angulata-, and
Bucklcl~di-beds (pars), arranged in a gentle syncline. At St. Mary's-
Well Bay, the Rh~etic again makes its appearance. North of
Lavernock Point the strata rise into a gentle anticline, with the
result that the Keuper makes its appearance. Thus Keuper,
lthmtic, and Lower Lias can all be examined ill this unrivalled
coast-section. Although brief reference has been made to the section by several
authors, 1 it has not, received sufficient attention. The most
complete record is that given by Bristow; but
he mentioned
comparatively few fossils. :For several reasons, a very detailed
section has been appended to tile present paper (facing p. 392). The Keuper :Red Marls are of the usual type: dark-red marls,
with greenish-grey zones. Above come the' Tea-Green Marls,'
having, according to my measurements, a total Thickness of 33 feet
4 inches. The most interesting feature in connection with these
marls is the occurrence in them of gypsum--a mineral that is
particularly rare ill North-West Gloucestershire and Worcestershire. Indeed, I have not found it in the marls of the sections which I
have studied in that district in any appreciable quantity. y
pp
q
y
Above the ' Tea-Green Marls ' come the Sully Beds--a portion of
Etheridge's 'Grey Marls.'
The line of demarcation between the
two series of deposits is necessarily an arbitrary one. The peculiar
lithic characters of the Sully Beds at Lavernock, and the fact
that John Storrie found '....some remains of the great Laby-
rinthodon, ~lctstoclonsaurus .... ', in association with a number
of small teeth of Sph~rodus, a mandible believed to belong to
Pal~eosa~trus, two teeth belonging to the same dinosaur, and
remains of Trematosaurz~s, about 6 feet below the Rhmtic ' fish-
bed,'" all support the conclusion that the Sully Beds are more
intimately connected with the Rh~etic than with the Keuper. The
most useful evidence in supporb of this contention, however, was
obtained at; St. Mary's-Well Bay (Sully), Cross, and Cadoxton;
as will be shown later (pp. 395, 396, & 399). a [Since this paper was written I have made a detailed examination of the
Watchet district ; and it appears desirable to state here that organic remains
are numerous in that district, in marlstone~ corresponding to these Sully
Beds.--dune 16th, 1905.] Vo]. 6 I.]
co~I6UOl~S DEPOSITS OF ~L~.MORGANSItIRE. 389 i. The Penarth District. In the Penarth district are included the outliers of Leckwith,
Penarth, Lavernock, and Cross:
all capped by Lower Liassic
deposits. The Penarth section, it is almost superfluous to state, is a classic
one, as the name of the locality suggested to Murchison an alternative
denomination for the Rh~etic--the Penarth Beds. It is unnecessary,
for the present purpose, to discuss all that has been written on this
far-famed section; but, for the convenience of those who desire to
study the literature, references are given in a footnote. 1 y
g
The cliff-section at Penarth is disappointing. It is true that there
is a faulted syneline which has brought the Rh~etie to the foot of
the cliff, but recently (August 1904) a slip has obscured the greater
part of the section which was available. Accoi-dingly, if anyone
wishes to obtain details of value, it is necessary (to quote Mr. H. B. Woodward) ' to climb an almost perpendicular cliff.'
Fragments of
the several hard beds can be seen on the beach, and ii'om an examina-
tion of their lithic structure it is often possible to state that such
and such a bed is preseut in the cliff-section ; but that is about all. The details recorded by Bristow and Etheridge were obtained in a
railway-cutting; and at the present time several such exposures
are available, especially at Penarth Docks. p
y
It is not necessary, however, to climb Penarth Head, for at Seven
Sisters' Bay the base of the Rh~ctic is within 3 feet of the beach ;
while at Lavernock Point the beds are even more accessible and
convenient to examine than at Garden Cliff, Westbury-on-Severn. The section at Seven Sisters' Bay is very similar to that at
Lavernock ; if anything, the beds arc a little thicker, and continue
to increase in thickness until Penarth Head is reached. At
Penarth Head :Mr. Woodward has noticed a band of limestone,
which reminded him of the Cotham Marble ; and I have observed
a similar bed--which probably represents the Sun-Bed of the White
Lias--at Lavernock (p. 393) and Barry (p. 398). On the beach at
Seven Sisters' Bay were pieces of l~mestone, probably from the
same horizon, that had been bored by Lithophagus (Lithodo~,~s,
Cuvier) and encrusted with Plicat~dct int~fs-striatct, Emmrich. 1 R. Etheridge, Trans. Cardiff ~Nat. See. vol. iii (1870-71) pt. it, p. 39
H. W. Bristow, (Geol. Surv.) Vert. Sect. Sheet 47. , (
)
Trans. Cardiff Nat. See. vol. xxvi (1893-94) pp. 105-106 & pl.
[Si
hi
i
I h
d
d
il d
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at University of Wisconsin -
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Downloaded from 1 R. Etheridge, Trans. Cardiff ~Nat. See. vol. iii (1870-71) pt. it, p. 39
H. W. Bristow, (Geol. Surv.) Vert. Sect. Sheet 47.
Trans. Cardiff Nat. See. vol. xxvi (1893-94) pp. 105-106 & pl.
a [Since this paper was written I have made a detailed examination of the
Watchet district ; and it appears desirable to state here that organic remains
are numerous in that district, in marlstone~ corresponding to these Sully
Beds.--dune 16th, 1905.] (A) Lavernock. ~ pp
The uppermost marlstone of the Sully Beds is conspicuously
waterworn, some of the irregularities projecting at least 5 inches Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from :Fig. 1.--View showi,,,l the upper s,~face of lhe Stdly Beds a~,d
tl~e Bbwlc Shales, Lave~'nocl~" Poi~,t. :Fig. 1.--View showi,,,l the upper s,~face of lhe Stdly Beds a~,d
tl~e Bbwlc Shales, Lave~'nocl~" Poi~,t. J. Storrie photogr. Fig. 2.--View showi~g the White Lias ee~(1 its ju~ction with the
Lower Liels at Lca.e~'~wclc Point. J~ Storrie photogr. J. Storrie photogr. J. Storrie photogr. Fig. 2.--View showi~g the White Lias ee~(1 its ju~ction with the
Lower Liels at Lca.e~'~wclc Point. Fig. 2. View showi g the White Lias ee (1 its ju ction with the
Lower Liels at Lca.e~'~wclc Point. J~ Storrie photogr. J~ Storrie photogr. J~ Storrie photogr. J~ Storrie photogr. Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from T.E Rtt.~ETIC DEPOSITS OF GLA~IORGANStIIRE. Vol. 6i.] Vol. 6i.] 391 above the average level (see fig. 1). Frequently a black-shale
deposit is found filling-in these irregularities ; but it is occasionally
replaced
by a soft yellowish-green marl, which passes up
gradually into a blackish marl. At that part of the cliff where
these notes were made, a layer of grey argillaceous nodules, with
quartz-pebbles and fish-remains, rested upon the above-mentioned
marl or shale. Doubtless this baud of nodules is intimately connected
with the ' fish-bed' for which the locality is celebrated. The fact
that black shale is intercalated between the' fish-bed' and Sully
Beds ill places, is commented upon by Mr. F. T. Howard, F.G.S.,
who, like most observers, had regarded the ' fish-bed' as the lowest
Rh~etic deposit in the Cardiff district. 'I was therefore,' he wrote, 'surprised, on removing a block of the con-
glomerate [' fish-bed'] from its natural position on the foreshore opposite
Lavernock Point, to see beneath it and lying directly on a worn surthee of
Tea-Green Marls, a thin band of black shale, not more than 88 inch thick,
containing numerous crushed specimens of typical Rhmtic shells--Cardium
rhceticum, Avicula contortct, and Pecle~ valoniensis. Others may have been
represented, but were badly preserved. The baud of shale was very
irregular .... 1 There are places where the ' fish-bed' rests directly upon the
marlstone of the Sully Beds. p
3 The teeth thus denominated are those described by Agassiz as Sphcerodus
minimus, and by Meyer & Plieninger as Psammodus orbicularis. They may
belong to the same animal (S, trgodon tomicus) as the chisel-shaped teeth, but at
present itappears desirable to keep the records separate. Charles Moore thought
that the knob-like teeth probably belonged to Lepidolus, and such teeth are
recorded in this paper as ' Sar.qodon tomicus' (Lepidotus ?). See Quart. Journ.
Geol. See. vol. xvii (1861) p. 499, footnote. i Trans. Cardiff Nat. Soc. vol xxix (1896-97) p. 66. Ibid. vol. xiv (1882-83) p. 100. MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, similar nature at Garden Cliff, Westbury-on-Severn. At Garden
Cliff, 16 inches above the basal bone-bed is the ' Lower Pullastm-
Sandstone.'
At Lavernock, 18 inches above the presumed equiva-
lent deposit, is a thin layer composed of hundreds of teeth of
Acrodus miuimus and scales of G'yrolelds Alberti. At Garden Cliff
a deposit of shale, 2 feet thick, separates the Lower from the Upper
Pullastra-Sandstone ; at Lavernock the deposit intervening between
the beds that may be regarded as the equivalents of the sandstones
measures only 13 ~nchcs. Bed 17 is also a 'bone-bed,' and frequently
contains vertebrae of l~'lesiosa~.us. Black shales with a grey lime-
stone-band separate this bed from a series of sandstone-layers
with shale-partings, and frequently full of fish-remains, usually
comminuted. If the correlation of the Rh~etic deposits (20 to 16)
enumerated above be correct, then the next bed in ascending order
should be the equivalent of the Bone-Bed (15) of Garden Cliff. This
was thought to be the case by Etheridge, and it certainly seems
probable,
ffohn Storrie wrote :-- similar nature at Garden Cliff, Westbury-on-Severn. At Garden
Cliff, 16 inches above the basal bone-bed is the ' Lower Pullastm-
Sandstone.'
At Lavernock, 18 inches above the presumed equiva-
lent deposit, is a thin layer composed of hundreds of teeth of
Acrodus miuimus and scales of G'yrolelds Alberti. At Garden Cliff
a deposit of shale, 2 feet thick, separates the Lower from the Upper
Pullastra-Sandstone ; at Lavernock the deposit intervening between
the beds that may be regarded as the equivalents of the sandstones
measures only 13 ~nchcs. Bed 17 is also a 'bone-bed,' and frequently
contains vertebrae of l~'lesiosa~.us. Black shales with a grey lime-
stone-band separate this bed from a series of sandstone-layers
with shale-partings, and frequently full of fish-remains, usually
comminuted. If the correlation of the Rh~etic deposits (20 to 16)
enumerated above be correct, then the next bed in ascending order
should be the equivalent of the Bone-Bed (15) of Garden Cliff. p
" Pte~4xt, Scopoli, = Avicula, Brug. I am at present engaged in the revision
of the representatives of the Pteriidm from the Rh~etic, Liassic, and Inferior-
Oolite reeks; and also those of the Mytilidm ti'om the same rooks. (A) Lavernock. As noticed by John Storrie, the ' fish-
bed' is not continuous, but occurs in patches, and the best of these
will be found protected by seaweed between low- and high-tide
marks :~ 'The patches in which it occurs vary from the size of a sixpence to pieces
5 or 6 yards square, and at some places are comparatively close together--at
others a considerable distance apart. The bed may be best described as a
conglomerate of pure white quartz-pebbles, from the size of a hen's egg down-
ward, all waterworn, and mostly crusted with a greenish-copper tinge, jasper-
like pebbles, and also some waterworn pieces of limestone (unfossiliferous, but
probably Silurian, from their general texture)."2 ]: am inclined to agree with Mr. Howard that the last-mentioned
pebbles are more likely to be of marlstone derived from the Sully
Beds. A fine piece of this 'fish-bed,' not altogether of the usual
lithic structure, since the greater mass of it consisted of quartz-sand
cemented together mainly by carbonate of lime, contained in great
abundance the teeth of Acrod~ts ~'ai~imus, Gyrolej~is Alberti (~P),
S~turichthys acun~i~atus, Sargodon tomicus, and Lepidotus (?),3 and
less commonly of Hybod~,s minor and H. cloaci~us. Rich as this
deposit is in vertebrate-remains, it has not been regarded by our
chief authorities as the Bone-Bed. It may be suggested that this
basal bone-bed at Lavernock is the equivalent of a deposit of Madison on June 20, 2016
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Downloaded from 392 MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, Trans. Cardiff Nat. See. voI. xiv (1882-83) p. 100.
" P
4
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at University of Wisconsin -
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Downloaded from MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, Ammonites in section. Ostrea liassica, 8trickland, aft• irregular@, Mtinster. 7
7
l
4
Modiola mi~dma, 8owerby non Moore. 2
Lima qiqautea (small). Psiloceras Joh~tstoni and radicles of Pseudodiadema in the nodules. 10
Psiloeeras planorbis : vertebrae of Ichthyosaurus. 7
Psilocera~ Joh~stoni (?). 2
~ t Ostrea lia~siea, Lima .qigaMea (Sowerby~, small; numerous radicles of
4
Pseudodiadema. 03 } Radicles of Pseudodioxlema. 9
11
Ossicles of Pentacrb~us. Ossieles of Pentacr6, us. Very small ossicles of a Peutm~inus, Lima (Pla qwstoma) sp. Lima (Cte~wstreon) Terquemi, Tats ; Peete~ (Chlamys). t Ostrea liassiea, common Pseudodiadema lobalum, Wright. t Ostrea liassica, common. 0
Pleuromya crowcombda, auett. [non Moore]. oi[
4
7
Ostrea li~sica, common. 6
3
0
Ostrea liazsiea. These beds are known locally as 'the Washers.'
4
Ostrea l~sica, abundant in the lower part; Modiola minima, Moore (large form). 4 t Pl&atula iatus-striata, Emmrieh; Ostrea llassiea; Lima valoniensis, Defra~ce. ¼) Modiola mUdma (large form); Plieatula hettangiensis, Terquem. Plieatula intu~-striata (common), Pleuromya (casts), Pleurophorus ? (casts),
O±
[ Pticatula hettanglensi~. 2 ~
Plieatula iMus-striata•
1
2
5
~
Alga (~). 4
uped with the Lower Lisa Thickness in ,feet
/ H~MI~RA ROTIFORMIS(?)
Limestone-beds and elay-pa~ings: estimated at ................................................ 25
HEMERA MARMORE.A~'. Marly days and nodular limestones: about... 40
I. Limestone ..................................................................................................... 0
2. Shale .............................................................................................................. 0
3. Limestone, nodular ....................................................................................... 0
t~ ~ Shale, with three beds of rather nodular limestone ............................................... I
m.j
11. Limestone, nodular : 0 to 2 inches ................................................................... 0
12
to ~ Shale, with three beds of limestone ................................................................... 2
~8.j
19. Limestone, massive : 4 to 8 inches ..................................................................... 0
20. Shale ............................................................................................................. 0
21. Limestone : 3 to 5 inches ................................................................................... o
22. Shale ........................................................ . ..................................................... i
WHITE
LIAS *
23. Limestone
....................................................................................................... 0
24, Shale ..... : ......................................................................................................... 0
25. Limestone, fissile ............................................................................................. 0
26. Shale .............................................................................................................. 1
27. Limestone, massive, with large nodules helow ; and beneath these are shales, with
numerous specimens of O~rea at the base ...................................................... 2
28. Limestone, massive ......................................................................................... o
,~0~ 8hale-par~ing, indistinct in places ........................................................................ o
Limestone, nodular .......................................................................................... 0
31• Shale ............................................................................................................... o
to
Limestones, five beds, intorstratifled with shale .................................................. 4
4o.j
41. Shale, hard, imperfectly laminated ..................................................................... 1
42. Limestone ......................................................................................................... 0
4:;. Shales, very much indurated in places, forming almost a limestone at the base ...... 2
44. Limestone, massive .......................................................................................... o
45. Shale-parting often indistinct, Bed 44 often adhering to 46 .................................... MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, 0
46. Limestone, somewhat nodular ........................................................................... 0
47. Shales ......................................................................................................... 0
48. Limestone, hard ............................................................................................ 0
49. Shales, with hard nodules ................................................................................ 1
50• Limestone, massive, very irregular surface, sometimes with nodules adhering to it... 0
51. Shales ............................................................................................................ 1
52• Limestone ...................................................................................................... 0
53. Shale : 3 to 5 inches ........................................................... .............................. 0
54. Limestone ...................................................................................................... 0
55. Shale : 2 inches ............................................................................................... )
56. Limestone, nodular, impersistent: 2 inches ......................................................... ~
0
57• Shale: 7 inches ................................................................................................ 58. Limestone, somewhat nodular ........................................................................... 0
59. Shales .......................................................................................................... 0
60. Limestone, nodular, impersistent ............................................................. ......... ()
61. Shales ........................................................................................................... 0
62. Limestone ...................................... ~ ............................................................... o
63. 8bales, bluish ................................................................................................... 1
64. Limestone, in two massive beds, but sometimes conjoined .................................... 0
65. Shales, bluish, with one or two layers of impure limestone ................................... 1
66. Limestone. .................................................................................................... 0
67. Shale-parting ................................................................................................... 0
68. Limestone ....................................................................................................... 0
69. 8hale or limestone ........ . ................................................................................. 0
70. Limestone ...................................................................................................... 0
71. Shale or limestone ......................................................................................... 0
72 ~
to
Limestones, five beds, with shale-partings ............................................................ 2
80.j
81. 8hales ............................................ : ............................................................... 0
82. Limestone ................................. ~..... ............................................................... 0
83. 8hale-parting ................................................................................................... 0
84. Limestone ........................................................................................................ 0
85. Shales ........................................................................................................... 0
86. Limestone .................................................................................................... 0
87. Shale, dark, thinly laminated ............................................................................. 0
88. Shale and thin limestone .................................................. ........................ : .... 1
89. Limestone, hard
............................................................................................. 0
90. Shale, hard, thinly laminated ........................................................................... 0
91. Limestone, in places i~iving way to a tough shale .............................................. 0
A2. aper-Shales,~hard, l~rown .................................................................... ..... 1
.:...%. • Shales, white or pale-grey, marly~ imperfectly laminated, with harder oanus in
B
places ......................................................................................................... 6
• Limestone, hard, grey, weathering into two beds, the upper one being probably the
representative of the Sun-Be-d . ................................................................. 0
[ 1. 8hale-parting: indistinct .............................................................................. 0
| 2. Limestone, hard, grey: half au inch to 2~ inches .......................................... 0
:. Shale, grey, par~ing ................................................................. : ........... 0
t 4. Limestone, greyish-green .............................................................................. 0
,~ ] 5. Shale, dark-grey ......................................................................................... ~" ~ 6. Limestone, greyish-green: 2 to 3 inches ......................................................... [ 7. Shale, hard, dark-g~ey with pyritic, gritty, ripple-marked layers .................... I 8. Limestone, very gntty, ripple-marke~
........................................................ 0
! .Q
--
. . . . ,
,. Shale analogous to 7, w~th snmlar gritty rapple-marked layers ........................ 0
( 10. Limestone, impure, gritty, ripple-marked : 3 to 6 inches ................................. 0
inches. 0
Montlivaltia [from a fallen block]. Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, Pteria (Avi~Tt/a) contorta, 5bhizodu~ :Ewaldi, Gervillia trr~cursoe, Quenstedt ; Proto-
8
cardium Philippianum, Peeteu valoniensis, Pleuromya (?), Modiola minima ; Orbi-
culoidea Townshendi (Davidson) ; fish-scales (Gyrolepi~). 8
Peeten valoniends. 8
Gyrolepis Alberti (scales). 10
Crowded with Pteria (Avic~la) eo~itorta along certain lines•
j Peegen ~aloniensis, Pteria (Avicula) eoatorta. (At Seven Sisters' Bay this bed is
4
nodular and from 0 to 8 inehes thick). Coproli~s. 0
8chizodus Ewahti, Pteria (Aviculo) conto~ta, and coprolites. (Beds 10 to 14 are
3 feet 8 inches thick at Seven Sisters' Bay.)
3
Ggrolepis Alberti (comminuted). 1
Coprolites, fish-scales (comminuted). l
4
Acrodus minimus, coprolites. 3
1
7
O~
Acrodu8 minimus, Gyrolepis Alberti. 1
Coprolites•
Sauriehthys aeuminatus. Aerodus miaimus, .G.wolepls Alberti, Hybodus minor, all
l
most abundant ; .vertebrae of Plesiosaurs, ieh thyodorulite of Hybodus, ' 8argodou
tomicu~' (see p. 391, faotn~te 3L
6
I Aerodus minim~, Ag., ,~aurichthw
acumiaatus,
Ag., 8argodon tomkms, Plieninger,
5
'S.t~,dcus"
(Learns?), Hybodus minor, Ag., H. doadnus, Quenstedt (rare),
ann Iragmen~s el Dories. 4~
}- See p. 389. 0)
1.' ~TEA-GREEN
MARLS.'
(33 feet 4 inches.)
II. Rzv IIARLS. i" Marlstone, rather broken up, yellowish-grey, thin layer of gypsum on the top ...... 0
9
. Green marl, much broken up, and containing near the top an irregular zone of
gypsum ................................................................................................... 1
4
3. Marhtone, greyish-yellow .............................................................................. Green marl, grey zoue near the top ................................................................. aaxlstone, hard, greyish-yellow, one of the most conspicuous beds in the section. 2
4
~
Marlstoue, laminated at the top ..................................................................... Greenish-grey marls, with hard zone near the centre ....................................... 2
5
Marlstone, earthy, with gypsum in cavities ...................................................... Greenish-grey marls ....................................................................................... 2
4
Series of t~ve bands of reddish marlstone, parted by thin, green, and less-compact
layers of marl. Some of the bands are not altogether red .................. .% ....... 4
2
[ 12. Greenish marl ................................................................................................ 0
6
I 13. Thin i.m~persistent band ofmarlstone ............................................................... 0
2
4
14. Greenisn marl ............................................................................................... 0
15• Marlstone
................................................................................................... 5
16. Greenish marl, somewhat laminated in places, compact at the base where it fits into
0
6
17. Deposit of gypsum, pink .............................................................................. 1010
~
18. Greeaish-grey and whitish marls, white, marl usually in hard layers .................. 19. Gypsum, usually white .................................................................................... {27~ Hard . . . . grey marl; 6 to 9 inches ........................................................................ O0
16l
Greemsh-grey marl, shaly m places, nodular m others ....................................... Red marls, with zones of greenish-grey marl. MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, / UPPER RHmTIC. \ LowER Raa~'r~c. 4. Marls, greyish-green. The gritty material fi'om the overlying bed has filtered down
cracks in this deposit ................................................................................... 2
/ 5 a. Shales, black ................................................................................................. 5
5h. Limestoue, sometimes in two layers with a very thin parting of sbaly matter. ' Beef' sometimes occurs above, and when the bed is in two layers it often occurs
between them ............................................................................................. 0
6. Shales, black, laminated .................................................................................... 1
7• Limestone. hard, bluish : 5 to 11 inches ............................................................... 0
8 a. Shales, black, laminated ................................................................................ 1
8 b. Shales, black, earthy. Beds 8 a and b are 4 feet thick at Seven Sisters' Bay ......... 1
9. Limestoue, hard, bluish, with one or twoshelly layers immediately below. Inplaces
the limestone becomes thin, and is then rephtced by grey earthy shales full of
shell-ddbris; 2 to 6 inches ........................................................................... 0
lO [ Shales, black, somewhat thinly laminated ............................................................ 2
2
I~4~:
~ Shales, hlack, tough, throwing, off water .............................................................. 1
[a. Limestone, grey, mlcaceous, arenaceous, and occasionally rel)laced by a more
|
shaly deposit with many 8dtizodl (?). the lower portion is often very pyritic
• |
and of bone-bed nature ........................................................................... t5"~b. Hard sandstone, slightly pyritic: I to 2 inches ................................................ 0
|c. 8hale-parting ................................................ i ............................................... 0
d. Two beds of sandstone,, with shale-parting and' beef' in between .......... .............. 0
a. Shales, black, laminated ................................................................................ o
16. ~ b. Limestone, dark grey, slightly pyritic ............................................................... o
e. Shales, black ................................................................................................ o
17. Thin' bone-bed,' intermittent: 0 to 1] inches ...................................................... 0
18. Shales, black ................................................................................................... 1
19. Thin 'bone-bed'. ............................................................................................... 0
20. Shales, black, laminated .................................................................................... 1
21. ' Fish-bed,' sometimes resting directly upon the top stratum of the Sully Beds, and at
others separated therefrom by black shale or marl .......................................... 0
t! . Two somewhat massive beds of hard greyish-blue marlstone, weathering yellowish. Upper surface extremely irregular ...................................................... ......... 1
. Series of blackish-blue marls, w~th harder zone~, passing gradually upwards into
harder irregular limestone ........................................................................ 6
Series of bluish limestones, with dark marly partings. The middle and most
i
"I
. . . . massl e bed is conglomeratw, and can be traced across the foreshore ............ 2
4 Series of blackish-blue and yellowish marls, with hard zones, two of which are near
~,x a~
.................. , ............................................................................ MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, the centre
4
9 { Pecten (CVhlamys) valoniensis, Defranee; Sehizodus Ewaldi; Protoc~rdium Philip-
pianum (Dunker) ; Cardium cloaoinum, Quenstedt; Cardinia ; lignite. 6
Pteria (Avicula) eontorta, Pectea valoaiensi~, Protocardium Philippianum. 3
Pleurop]wrus (casts). Pteria (Avi~Tt/a) contorta, 5bhizodu~ :Ewaldi, Gervillia trr~cursoe, Quenstedt ; Proto-
8
cardium Philippianum, Peeteu valoniensis, Pleuromya (?), Modiola minima ; Orbi-
culoidea Townshendi (Davidson) ; fish-scales (Gyrolepi~). 8
Peeten valoniends. 8
Gyrolepis Alberti (scales). 10
Crowded with Pteria (Avic~la) eo~itorta along certain lines•
j Peegen ~aloniensis, Pteria (Avicula) eoatorta. (At Seven Sisters' Bay this bed is
4
nodular and from 0 to 8 inehes thick). Coproli~s. 0
8chizodus Ewahti, Pteria (Aviculo) conto~ta, and coprolites. (Beds 10 to 14 are
3 feet 8 inches thick at Seven Sisters' Bay.)
3
Ggrolepis Alberti (comminuted). 1
Coprolites, fish-scales (comminuted). l
4
Acrodus minimus, coprolites. 3
1
7
O~
Acrodu8 minimus, Gyrolepis Alberti. 1
Coprolites•
Sauriehthys aeuminatus. Aerodus miaimus, .G.wolepls Alberti, Hybodus minor, all
l
most abundant ; .vertebrae of Plesiosaurs, ieh thyodorulite of Hybodus, ' 8argodou
tomicu~' (see p. 391, faotn~te 3L
6
I Aerodus minim~, Ag., ,~aurichthw
acumiaatus,
Ag., 8argodon tomkms, Plieninger,
5
'S.t~,dcus"
(Learns?), Hybodus minor, Ag., H. doadnus, Quenstedt (rare),
ann Iragmen~s el Dories. 4~
}- See p. 389. 0)
1.' ~TEA-GREEN
MARLS.'
(33 feet 4 inches.)
II. Rzv IIARLS. i" Marlstone, rather broken up, yellowish-grey, thin layer of gypsum on the top ...... 0
9
. Green marl, much broken up, and containing near the top an irregular zone of
gypsum ................................................................................................... 1
4
3. Marhtone, greyish-yellow .............................................................................. Green marl, grey zoue near the top ................................................................. aaxlstone, hard, greyish-yellow, one of the most conspicuous beds in the section. 2
4
~
Marlstoue, laminated at the top ..................................................................... Greenish-grey marls, with hard zone near the centre ....................................... 2
5
Marlstone, earthy, with gypsum in cavities ...................................................... Greenish-grey marls ....................................................................................... 2
4
Series of t~ve bands of reddish marlstone, parted by thin, green, and less-compact
layers of marl. Some of the bands are not altogether red .................. .% ....... 4
2
[ 12. Greenish marl ................................................................................................ 0
6
I 13. Thin i.m~persistent band ofmarlstone ............................................................... 0
2
4
14. Greenisn marl ............................................................................................... 0
15• Marlstone
................................................................................................... 5
16. Greenish marl, somewhat laminated in places, compact at the base where it fits into
0
6
17. Deposit of gypsum, pink .............................................................................. 1010
~
18. Greeaish-grey and whitish marls, white, marl usually in hard layers .................. 19. MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, Shales, very much indurated in places, forming almost a limestone at the base ...... 44. Limestone, massive .......................................................................................... 45. Shale-parting often indistinct, Bed 44 often adhering to 46 .................................... 46. Limestone, somewhat nodular ........................................................................... 47. Shales ......................................................................................................... 48. Limestone, hard ............................................................................................ 49. Shales, with hard nodules ................................................................................ 50• Limestone, massive, very irregular surface, sometimes with nodules adhering to it... 51. Shales ............................................................................................................ 52• Limestone ...................................................................................................... 53. Shale : 3 to 5 inches ........................................................... .............................. 54. Limestone ...................................................................................................... 55. Shale : 2 inches ............................................................................................... )
56. Limestone, nodular, impersistent: 2 inches ......................................................... ~
57• Shale: 7 inches ................................................................................................ 58. Limestone, somewhat nodular ........................................................................... 59. Shales .......................................................................................................... 60. Limestone, nodular, impersistent ............................................................. ......... 61. Shales ........................................................................................................... 62. Limestone ...................................... ~ ............................................................... 63. 8bales, bluish ................................................................................................... 64. Limestone, in two massive beds, but sometimes conjoined .................................... 65. Shales, bluish, with one or two layers of impure limestone ................................... 66. Limestone. .................................................................................................... 67. Shale-parting ................................................................................................... 68. Limestone ....................................................................................................... 69. 8hale or limestone ........ . ................................................................................. 70. Limestone ...................................................................................................... 71. Shale or limestone ......................................................................................... 72 ~
to
Limestones, five beds, with shale-partings ............................................................ 80.j
81. 8hales ............................................ : ............................................................... 82. Limestone ................................. ~..... ............................................................... 83. 8hale-parting ................................................................................................... 84. Limestone ........................................................................................................ 85. Shales ........................................................................................................... 86. Limestone .................................................................................................... 87. Shale, dark, thinly laminated ............................................................................. 88. Shale and thin limestone .................................................. ........................ : .... 89. Limestone, hard
............................................................................................. 90. Shale, hard, thinly laminated ........................................................................... 91. Limestone, in places i~iving way to a tough shale .............................................. A2. aper-Shales,~hard, l~rown .................................................................... ..... .:...%. • Shales, white or pale-grey, marly~ imperfectly laminated, with harder oanus in
B
places ......................................................................................................... • Limestone, hard, grey, weathering into two beds, the upper one being probably the
representative of the Sun-Be-d . ................................................................. [ 1. 8hale-parting: indistinct .............................................................................. | 2. Limestone, hard, grey: half au inch to 2~ inches .......................................... :. Shale, grey, par~ing ................................................................. : ........... t 4. Limestone, greyish-green .............................................................................. ,~ ] 5. Shale, dark-grey ......................................................................................... ~" ~ 6. Limestone, greyish-green: 2 to 3 inches ......................................................... [ 7. Shale, hard, dark-g~ey with pyritic, gritty, ripple-marked layers .................... I 8. Limestone, very gntty, ripple-marke~
........................................................ ! .Q
--
. . . . ,
,. Shale analogous to 7, w~th snmlar gritty rapple-marked layers ........................ ( 10. Limestone, impure, gritty, ripple-marked : 3 to 6 inches ................................. Thickness in ,feet inches. / UPPER RHmTIC. \ LowER Raa~'r~c. 4. Marls, greyish-green. The gritty material fi'om the overlying bed has filtered down
cracks in this deposit ................................................................................... MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, 2
/ 5 a. Shales, black ................................................................................................. 5
5h. Limestoue, sometimes in two layers with a very thin parting of sbaly matter. ' Beef' sometimes occurs above, and when the bed is in two layers it often occurs
between them ............................................................................................. 0
6. Shales, black, laminated .................................................................................... 1
7• Limestone. hard, bluish : 5 to 11 inches ............................................................... 0
8 a. Shales, black, laminated ................................................................................ 1
8 b. Shales, black, earthy. Beds 8 a and b are 4 feet thick at Seven Sisters' Bay ......... 1
9. Limestoue, hard, bluish, with one or twoshelly layers immediately below. Inplaces
the limestone becomes thin, and is then rephtced by grey earthy shales full of
shell-ddbris; 2 to 6 inches ........................................................................... 0
lO [ Shales, black, somewhat thinly laminated ............................................................ 2
2
I~4~:
~ Shales, hlack, tough, throwing, off water .............................................................. 1
[a. Limestone, grey, mlcaceous, arenaceous, and occasionally rel)laced by a more
|
shaly deposit with many 8dtizodl (?). the lower portion is often very pyritic
• |
and of bone-bed nature ........................................................................... t5"~b. Hard sandstone, slightly pyritic: I to 2 inches ................................................ 0
|c. 8hale-parting ................................................ i ............................................... 0
d. Two beds of sandstone,, with shale-parting and' beef' in between .......... .............. 0
a. Shales, black, laminated ................................................................................ o
16. ~ b. Limestone, dark grey, slightly pyritic ............................................................... o
e. Shales, black ................................................................................................ o
17. Thin' bone-bed,' intermittent: 0 to 1] inches ...................................................... 0
18. Shales, black ................................................................................................... 1
19. Thin 'bone-bed'. ............................................................................................... 0
20. Shales, black, laminated .................................................................................... 1
21. ' Fish-bed,' sometimes resting directly upon the top stratum of the Sully Beds, and at
others separated therefrom by black shale or marl .......................................... 0
t! . Two somewhat massive beds of hard greyish-blue marlstone, weathering yellowish. Upper surface extremely irregular ...................................................... ......... 1
. Series of blackish-blue marls, w~th harder zone~, passing gradually upwards into
harder irregular limestone ........................................................................ 6
Series of bluish limestones, with dark marly partings. The middle and most
i
"I
. . . . massl e bed is conglomeratw, and can be traced across the foreshore ............ 2
4 Series of blackish-blue and yellowish marls, with hard zones, two of which are near
~,x a~
.................. , ............................................................................ the centre
4
9 { Pecten (CVhlamys) valoniensis, Defranee; Sehizodus Ewaldi; Protoc~rdium Philip-
pianum (Dunker) ; Cardium cloaoinum, Quenstedt; Cardinia ; lignite. 6
Pteria (Avicula) eontorta, Pectea valoaiensi~, Protocardium Philippianum. 3
Pleurop]wrus (casts). MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, Gypsum, usually white .................................................................................... {27~ Hard . . . . grey marl; 6 to 9 inches ........................................................................ O0
16l
Greemsh-grey marl, shaly m places, nodular m others ....................................... Red marls, with zones of greenish-grey marl. 4. Marls, greyish-green. The gritty material fi'om the overlying bed has filtered down
cracks in this deposit ................................................................................... 9 { Pecten (CVhlamys) valoniensis, Defranee; Sehizodus Ewaldi; Protoc~rdium Philip-
pianum (Dunker) ; Cardium cloaoinum, Quenstedt; Cardinia ; lignite. 6
Pteria (Avicula) eontorta, Pectea valoaiensi~, Protocardium Philippianum. 3
Pleurop]wrus (casts). Pteria (Avi~Tt/a) contorta, 5bhizodu~ :Ewaldi, Gervillia trr~cursoe, Quenstedt ; Proto-
8
cardium Philippianum, Peeteu valoniensis, Pleuromya (?), Modiola minima ; Orbi-
culoidea Townshendi (Davidson) ; fish-scales (Gyrolepi~). 8
Peeten valoniends. 8
Gyrolepis Alberti (scales). 10
Crowded with Pteria (Avic~la) eo~itorta along certain lines•
j Peegen ~aloniensis, Pteria (Avicula) eoatorta. (At Seven Sisters' Bay this bed is
4
nodular and from 0 to 8 inehes thick). Coproli~s. 0
8chizodus Ewahti, Pteria (Aviculo) conto~ta, and coprolites. (Beds 10 to 14 are
3 feet 8 inches thick at Seven Sisters' Bay.)
3
Ggrolepis Alberti (comminuted). 1
Coprolites, fish-scales (comminuted). l
4
Acrodus minimus, coprolites. 3
1
7
O~
Acrodu8 minimus, Gyrolepis Alberti. 1
Coprolites•
Sauriehthys aeuminatus. Aerodus miaimus, .G.wolepls Alberti, Hybodus minor, all
l
most abundant ; .vertebrae of Plesiosaurs, ieh thyodorulite of Hybodus, ' 8argodou
tomicu~' (see p. 391, faotn~te 3L
6
I Aerodus minim~, Ag., ,~aurichthw
acumiaatus,
Ag., 8argodon tomkms, Plieninger,
5
'S.t~,dcus"
(Learns?), Hybodus minor, Ag., H. doadnus, Quenstedt (rare),
ann Iragmen~s el Dories. 4~
}- See p. 389. 0) 8
Gyrolepis Alberti (scales). 0
Crowded with Pteria (Avic~la) eo~itorta along certain lines• Peegen ~aloniensis, Pteria (Avicula) eoatorta. (At Seven Sisters' Bay this bed is
nodular and from 0 to 8 inehes thick). Coproli~s. 8chizodus Ewahti, Pteria (Aviculo) conto~ta, and coprolites. (Beds 10 to 14 are
3 feet 8 inches thick at Seven Sisters' Bay.) 1.' ~TEA-GREEN
MARLS.'
(33 feet 4 inches.)
II. Rzv IIARLS. i" Marlstone, rather broken up, yellowish-grey, thin layer of gypsum on the top ...... 0
9
. Green marl, much broken up, and containing near the top an irregular zone of
gypsum ................................................................................................... 1
4
3. Marhtone, greyish-yellow .............................................................................. Green marl, grey zoue near the top ................................................................. aaxlstone, hard, greyish-yellow, one of the most conspicuous beds in the section. 2
4
~
Marlstoue, laminated at the top ..................................................................... Greenish-grey marls, with hard zone near the centre ....................................... MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, 0
Cardinia ovalis (Stutchbury), common in places. These beds are locally known as
the' Lavernock Shales' (angu/ata-zone). 2
3
6
Lima (Plgqiostoma) #~.antea~ Psiloceras J'ohnstoni (Sowerby). 8
Psiloeeras sp,
I
Psiloesras Johastoni. Ammonites in section. Ostrea liassica, 8trickland, aft• irregular@, Mtinster. 7
7
l
4
Modiola mi~dma, 8owerby non Moore. 2
Lima qiqautea (small). Psiloceras Joh~tstoni and radicles of Pseudodiadema in the nodules. 10
Psiloeeras planorbis : vertebrae of Ichthyosaurus. 7
Psilocera~ Joh~stoni (?). 2
~ t Ostrea lia~siea, Lima .qigaMea (Sowerby~, small; numerous radicles of
4
Pseudodiadema. 03 } Radicles of Pseudodioxlema. 9
11
Ossicles of Pentacrb~us. Ossieles of Pentacr6, us. Very small ossicles of a Peutm~inus, Lima (Pla qwstoma) sp. Lima (Cte~wstreon) Terquemi, Tats ; Peete~ (Chlamys). t Ostrea liassiea, common Pseudodiadema lobalum, Wright. t Ostrea liassica, common. 0
Pleuromya crowcombda, auett. [non Moore]. oi[
4
7
Ostrea li~sica, common. 6
3
0
Ostrea liazsiea. These beds are known locally as 'the Washers.'
4
Ostrea l~sica, abundant in the lower part; Modiola minima, Moore (large form). 4 t Pl&atula iatus-striata, Emmrieh; Ostrea llassiea; Lima valoniensis, Defra~ce. ¼) Modiola mUdma (large form); Plieatula hettangiensis, Terquem. Plieatula intu~-striata (common), Pleuromya (casts), Pleurophorus ? (casts),
O±
[ Pticatula hettanglensi~. 2 ~
Plieatula iMus-striata•
1
2
5
~
Alga (~). 4
* Th
Whi
Li
i
i i
ll
d
i h h
L
Li Thickness in ,fe
/ H~MI~RA ROTIFORMIS(?)
Limestone-beds and elay-pa~ings: estimated at ................................................ 2
HEMERA MARMORE.A~'. Marly days and nodular limestones: about... 4
I. Limestone ..................................................................................................... 2. Shale .............................................................................................................. 3. Limestone, nodular ....................................................................................... t~ ~ Shale, with three beds of rather nodular limestone ............................................... m.j
11. Limestone, nodular : 0 to 2 inches ................................................................. 12
to ~ Shale, with three beds of limestone ................................................................. ~8.j
19. Limestone, massive : 4 to 8 inches ..................................................................... 20. Shale ............................................................................................................. 21. Limestone : 3 to 5 inches ................................................................................... 22. Shale ........................................................ . ..................................................... WHITE
LIAS *
23. Limestone
....................................................................................................... 24, Shale ..... : ......................................................................................................... 25. Limestone, fissile ............................................................................................. 26. Shale .............................................................................................................. 27. Limestone, massive, with large nodules helow ; and beneath these are shales, with
numerous specimens of O~rea at the base ...................................................... 28. Limestone, massive ......................................................................................... ,~0~ 8hale-par~ing, indistinct in places ........................................................................ Limestone, nodular .......................................................................................... 31• Shale ............................................................................................................... to
Limestones, five beds, intorstratifled with shale .................................................. 4o.j
41. Shale, hard, imperfectly laminated ..................................................................... 42. Limestone ......................................................................................................... 4:;. MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, This
was thought to be the case by Etheridge, and it certainly seems
probable,
ffohn Storrie wrote :-- ' The bon~bed proper.., is a continuous bed through the whole [Penarth-
Laveruock] section, and is described by Etheridge as "a dark-grey grit, or hard
indurated pyritic limestone, 2 or 3 inches thick (oftener about 1 inch), made up
of m.inutely-comminuted fragments of fish-teeth, scales, and bones." This bed
... is accessible in the whole Penarth section, and is always constant in
character, except at Lavernock, where it is more pyritized than elsewhere. This bed very rarely contains large bones or spines ; on one occasion only have
I found such.' 1 The most persistent strata in the Pteria (Avicula)-contorta Zone are
the Pecten-Beds (7 & 5 b), especially the lower of the two. At
Lavernock, they can be easily traced across the foreshore. Bristow
observed that the lower bed was full of Pecten (Chlamys) valoniensis;
but, according to my investigations, it was of rather rare occurrence
in the actual limestone, and in neither of the Pecten-Beds did I find
lamellibranchs to be abundant, as is usually the case. However, the
Black Shales, for 2 or 3 inches below Bed 5 b, are extremely fossili-
ferous, containing Pecte~ ( Chlamyg) valonie~sis, Pteria (Avicula)
contorta, 2 scales of Gyrolepis Alberti, and coprolites (of fishes chiefly). In places immediately below this shale is a thin earthy limestone-
layer with a layer of ' beef' on the under surface. At 5 inches
below 5 b is a seam full of Pteria (Avic~tht) contorts ; and in the shales
below that again, numerous examples of Pecten (Chlamys) valoniensis,
and Schizodus Ewaldi, Gervillia prcecursor, Protocardium Philippi-
anum, Orbiculoidea Townshendi, Pleui'omya (:P ), Placu~wpsis alpina
(small and large forms), scales of Gyrolepis .Alberti, teeth of Acted us
minimus (rare) and Saurichthys acuminatus, and coprolites. The
vertebrate-remains occur mostly in a thin laver at 8 inches below 5b. This is certainly the most fossiliferous horizon in the Rh~etic at this
locality; and fortunately the deposit is easy to investigate. Black Quart. J0urn. (]sol 8o~., Aug. 1905.] [To face p. 392. SECTION AT LAVERNOCK~ NEAR CARDIFF. SECTION AT LAVERNOCK~ NEAR CARDIFF. hes. 0
Montlivaltia [from a fallen block]. 0
Cardinia ovalis (Stutchbury), common in places. These beds are locally known as
the' Lavernock Shales' (angu/ata-zone). 2
3
6
Lima (Plgqiostoma) #~.antea~ Psiloceras J'ohnstoni (Sowerby). 8
Psiloeeras sp,
I
Psiloesras Johastoni. CO~TmVOUS DEVOSITS OF GLA~O~GA~'SrnR~ shales complete the Lower Rha~tic Stage, and pass up gradually into
a greyish-green marl. This marl-deposit was much fissured, previous to the deposition of
the superincumbent gritty limestone. Into these fissures gritty
material was washed. We have here a non-sequence
caused by
a slight upheaval; and, as a result, certain beds which are found
elsewhere, are not to be seen at Lavernoek. The beds that
are missing at Lavernock measure at, Garden Cliff about 13~ feet. Gradual subsidence, again, in the Lavernock area allowed of the
formation of the White Lias (fig. 2, p. 390). At Penarth, Etheridge
failed to discover ' Ostrea intus-striata, so common in Somersetshire
and Warwickshire' ; ~ but at Lavernoek it is not uncommon, and is
very abundant at Coldknap, Barry. The probable equivalent of the
Sun-Bed completes the series of limestone-beds; and i~ is of this
stratum that 1 found bored pieces (crypts of Lithophagus), on the
beach at Seven Sisters' Bay. The specimens of Ostrea liassica
(two) from this stratum were on the upper surface. The light-
coloured shales above the White-Lias limestones are grouped with
the White Lias; they contain Ostrea liassica abundantly in the
lower portion. These are capped by the Paper-Shales, concerning
which there has been some debate as to whether they should be
classed with the Rhaetie or with the Lias.-"
I think that there is
little doubt but that they should be grouped with the Lias. The deposit between these shaly beds and the Lavernock Shales
requires no particular comment ; but there has been some doubt as
to the exact age of the last-named. However, as surmised by
:Mr. H. B. Woodward, they belong to the anelulata-zone, having
been laid down during the hemcra marmore~. Ccwdinia ovalis
(Stutchbury) did not exist after this hemera. ]n the Lavernock
Shales it is abundant at certain horizons, both at Lavernock and at
Leckwithbridge, near Cardiff. At the latter locality Sch?otheiraia
antjulatc~ (Sehlotheim) and Cardi,~ia ovalis occur in association;
consequently there can be no doubt as to the date of the beds. As
the Lavernock Shales are difficult to examine minutely at the typical
locality, the section at Leckwithbridge is appended :~ 77dckness i~l feet inches. :Limestone, two beds mixed with
marly clay ............... about J 0
8
Clay, marly, blue and yellow ... 0
8
Limestone ........................... 0
2
Clay, marly : 4 to 6 inches ...... (
) p
, p
2 H. ]3. Woodward, 'The Jurassic Rocks of Britain' Mere. Geol. Surv.
vol. iii (1893) p. 119. 1 Trans. Cardiff Nat. Soe. vol. iii (1870-71) pt. it, p. 50.
2 H. ]3. Woodward, 'The Jurassic Rocks of Britain' Mere. Geol. Surv.
vol. iii (1893) p. 119. 1 Trans. Cardiff Nat. Soe. vol. iii (1870-71) pt. it, p. 50. MR. L. RICHARDSON ON THE l~Hm~rlc AND
[Aug. 19o5, 2
5
Marlstone, earthy, with gypsum in cavities ...................................................... Greenish-grey marls ....................................................................................... 2
4
Series of t~ve bands of reddish marlstone, parted by thin, green, and less-compact
layers of marl. Some of the bands are not altogether red .................. .% ....... 4
2
[ 12. Greenish marl ................................................................................................ 0
6
I 13. Thin i.m~persistent band ofmarlstone ............................................................... 0
2
4
14. Greenisn marl ............................................................................................... 0
15• Marlstone
................................................................................................... 5
16. Greenish marl, somewhat laminated in places, compact at the base where it fits into
0
6
17. Deposit of gypsum, pink .............................................................................. 1010
~
18. Greeaish-grey and whitish marls, white, marl usually in hard layers .................. 19. Gypsum, usually white .................................................................................... {27~ Hard . . . . grey marl; 6 to 9 inches ........................................................................ O0
16l
Greemsh-grey marl, shaly m places, nodular m others ....................................... Red marls, with zones of greenish-grey marl. Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from Vo]. 6t.] 393 CO~TmVOUS DEVOSITS OF GLA~O~GA~'SrnR~ Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from CO~TmVOUS DEVOSITS OF GLA~O~GA~'SrnR~ 0
5
:Limestone ........................... 0
3 (
Clay and limestone-bands ...... 1
10
:Limestone, dark grey ............ 0
3
Clay, hard,marly, reddish blotches 0
5
Ostrea irregularis, Lima
( Plagiostoma )
aft. giyantea. Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from 394 MR. L. RICHARDSON ON THE RH.~-TIC AND
[Aug. t9o 5, (
,d
r
[Leckwithbridge Section (eontimted).]
Thickness in .feet inches. Limestone : .......................... 0
Clay, with three bands of lime-~
stone, totalliug 7 inches ...... ~ 4
Limestone ........................... 0
Clay and limestones ............... 12
Clay, blue ;
C~o'dinia
ov.[i~ ] 3
a'bundant ....................... f
Clay a~d limestones. Some of]
the limestone-bands become 1 2'2
nodular ..................... seen
4
Oslre. irreyularis. 0 ( O.~t~'ea irregularis, Lima
yiganleu ,
6'(trdiMa
3
ocalis,
ScMotheimia
a~gulata (Schloth.). Ca~'dinia ocaii~ (Stutch-
burs) : ()~lrea irref/u-
lari,s, Mfmster ; Lima. qiqanh'a
(Sowerby) ;
0 ! ~i'holadomya
.fort~t. m~la,
Dumortier ;
0,~ "
Litlorina cf. "mi~tuta,
t
Terq. & Piette ; ossicles
ofPe~t lacri~ u~,radioles
4
of Cidaris ; Rh.qncho-
nclla aft. calcicosta,
Day. ; Dentalium ela-
lense, Terq. & Piette. [Leckwithbridge Section (eontimted).]
Thickness in .feet inches. Concerning the higher beds at Lavernock no details can be given ;
t, hey are inaccessible, but limestones predominate. Proceeding
farther westward along the coast, the other side of the syneline is
entered upon, and the strata are soon passed over in descending
order, until at St. Mary's-Well Bay, Sully, the Rh~etic reappears. 3 , The Geology of the South-Wales Coalfield: Pt. iii' Mere. Geol. 8urv.
1902, p. 63. (B) St. Mary's-Well Bay, Sully. At this locality the whole of the Rhmtie Series can be studied ;
but, since the Lavernoek section has been dealt with in Such detail,
it will be sufficient for our present purpose to describe the basal
portion
only of the exposure. The beds are very much disturbed,
owing to a fault--that which starts at Dinas Powis. The result is
that the Rh~etic Beds are faulted against the littoral Keuper, and
iu the downward course that portion of the series which I have
denominated the Sully Beds has been prettily contorted. y
p
y
The Bone-Bed seen in this section, although consisting of several
layers, as at Lavernock, is nevertheless lithically distinct. The main
band is a hard grey limestone, seldom pyritic, but usually crowded
with fish-remains, although no quartz-pebbles were observed. :Below
the Bone-Bed are black shales with intermittent hard layers, as
noticed in the appended section :-- Thic],'ne~s in./'eel im'hes. 15. A series of sandstone- aud lime- ]
stone-layers, with partiugs i 1
of shale ........................... 16. Shales, black ........................ 1
17. Limestone; intermittent ........ 0
18. Shales, black ........................ 0
19. Limestone, earthy ; intermittent 0
20. Shales, black ........................ 1
21. l~ust-eoloured layer ............... 0
( Gyrolepis Alberti (teeth ? &
scales) ; a lamellibraneh
0 ]
(Schizodu.s o1" Pullastra)
L
not u]lcomlllon
1
5
1
4
O~ Gyrole#is Alberti (rare). Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from Thickness in feet inches. Thickness in feet inches. ( Marlstone, hard, greenish-grey, ]
( Ostrea Bri~toci (very com-
mon) ; l'/eria (AvicubO
I
massive
at
the top, butj. 4
6~ ~ contorts (abundant in a
~
nodular and mixed with black
I
and yellow marlbelow ....... I
thin layer at the top);
Marls and marlstoues. I
&~rgodon (?),~ Le~)idotus
~
(?) ; 3lodiola. Large masses of marlstone from the Sully Beds, scattered about
on the beach, are crowded with Ostrea Bristovi, Etheridge, MS. (see
p. 422), and the beds also frequently contain traces of the mineral
baryto-celestine. These strata do not appear to have attracted any
attention ; they must have been observed, because Mr. E. T. INewton,
FIR.S, informed me that an O, trea, similar to that which I had
submitted to him for examination from this locality, was preserved
in the Museum of Practical Geology, Jermyn Street, and bore the
legend ' Ostrea Bristovi, Etheridge, MS. From near l~enarth. '
In a
thin layer at the top of the Sully Beds, Pteria (Avicula) contorts is
very abundant.:
Their fossil contents render it incumbent that
these beds should be classed with the Rhmtic, but an arbitrary line
of division mush be drawn between them and the ' Tea-Green Marls,'
which will be subject to alteration according to the records of
fossils. (C) Cross Farm,
near Dinas
fowls. Pyenodont fish. It is preserved ]n the collection of Mr. E. Talbot Paris.
o The abundance of Pteria contorta iu the uppermost layer of the Sully
:Beds here, and the record by Mr. F. T. tIoward, F.G.S., of' numerous crushed
specimens of' typical RhwAic shells--C(o.dittm r)teeticum. Ach'ula cow, terra, and
Pecten valoniensis '--in a thin layer of shale below the 'fish-bed,' Trans. Cardiff
,Nat. Soc. vol. xxix (1896-97) p. 66, is interesting in connection with a statement
made by Prof'. S. IT. Reynolds & Dr. A. Vaughan in a [botnote to their admirable
paper on 'The Rh~etic Beds of the South-Wales Direct Line' Quart. Journ.
Geol. See. vol. lx (1904) p. 200.
In that footnote they remarked :--' We have
ventured to dissent somewhat from Mr. Richardson's correlation of the beds
at Garden Cliff. Seeing that Avicula contorta and Schizodus occur plentifully
below his Bone-Bed (:Bed 15), it does not appear to us that this bed can be
considered to be on the same horizon as that at Sodbury, which is well below
the levelat which these mollusca commence to occur in any abundance.' As to
whether the ~odbury :Bone-:Bed was on the same horizon as that numbered
15 at Garden Cliff was a question for them to decide, and the evidence which
led to their answer in the negative is quoted above.
In xiew of the details
obtained in Glamorganshire (and also iu the Watcher district), this hardly
seems conclusive. Vol. 6x.]
COSITIGUOUS :DEPOSITS OF 6LAMOR6ANSItlRE. 395 ~,R. T,. R,Cm*RDSO.~ O.X THn R~a~TIC A~D
[Aug. ,905, ~,R. T,. R,Cm*RDSO.~ O.X THn R~a~TIC A~D
[Aug. ,905, and 1896, a large quarry has been opened out in the Lower Liassic
limestones of this outlier, near Cross Farm, and the junction of
these beds with the White Lias (A of Lavernock) exposed to view. The section is as follows :~ WIIITE 1
LIAs. Shales, bluish, marly ............ seen
Thick,tess in fi,et
:Marl and limestone-fragments ...... seen
0
:[hr e bands of hmestone, with marly~ o
partings ....................................... )
Limestone ...................................... 0
Limestones and shales ....................... 5
Shale ............................................ 0
Limestone in two conspieuolks beds ...... 0
Shale ............................................ 0
Limestone ....................................... 0
Limestone and shales ........................ 9
Three beds of limestone, with very thin]
partings of shale. Locally called ' the / 0
Washers' . .................................. Clay ............................................. 0
Limes~,one ....................................... 0
Shale, thinly laminated in the upper por- 1 0
tion, clayey in the lower ................ j'
Limestone, blue-centred ..................... 0
~ Paper-Shales, as at Lavernock ............ 1
3
Dwhes. 10
lO
Oslrea liassica and
7
radioles o[ Pseudo-
dh~dema. 2
4
Ostrea liassica
eom lnoll. 5
5
7
Ostrea liassiea. 1
6
Oslrea [iassica. 6
3
0
6 Unfortunately, t.he sequence downwards
cannot be ascertained;
but in the lane-side near the buildings at Cross Farm the following
details may be observed :- Thickness in feet bwhes. Shales, black, with soft gritty layers at
(In the limestone Pul. the base
........................... seen
3
0
htstra arenicola is
abundant: wood and
15. Limestone, hard, grey, mica~ous,
fragments of fish-
with
quartz-~nd
immediately
scales occur. In the
below ................................... 0
5! quartz-rand Acrodus
minimus,
Saurich-
l h ys az~tminatus, and
G yrolepis Albert i are
16. Shales, black ........................... 0
4 ~ common. 17. Sandy layer, choeolate-coloured ... 0
l
Ggrolepis Alberti. [ Gap ]
{'Marlstone, hard, greenish-grey: 4 to
~ [
15 inches ................................. 0
9
O~trea Bristovi. ;~ ~ Marly shales, dark greenish-grey and
oz
I
brown, with sandy seams ............... 2
0
[ Marlstone, hard, greenish-grey ......... 0
11
Ostrea Jgristovi. Thickness in feet bwhes. The lamellibranchs in Bed 15 are, on the whole, well-preserved
and extremely abundant. As I did not recognize the fossil, I
submitted specimens to Mr. E. T. (C) Cross Farm,
near Dinas
fowls. In the memoir descriptive of the geology of the Cardiff district
it is observed that ' To the west of the Lower Penarth or Lavernock outlier there is a consider-
able tract just high enough to take in some of the Rhmtic shales and limestones,
but nowhere high enough to touch the Lias. '3 Since the district was geologically surveyed between the years 1892 l Concerning this specimen, Dr. A. Smith Woodward, F.R.S., wrote (in. lilt.):--' Premaxilla with cutting-teeth. Might be S(oyodon, or perhaps a
Pyenodont fish.' It is preserved ]n the collection of Mr. E. Talbot Paris. h
b
d
f
i
h
l
f
h
S ll Madison on June 20, 2016
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Downloaded from 396 ii. Barry to Cowbridge. In the district between Barry and Cowbridge there are three
important sections of the beds under consideration, namely, at
Barry (Coldknap), Cadoxton, and Tregyff (near Cowbridge). y
p
gy
g
Two small outliers of Rh~etic Beds occur on Barry Island. The
northernmost patch has been investigated by Mr. F. T. Howard, :F.G.S.,
who has recorded a number of fossils.: are found to have changed, both as regards faunal and lithic
characters :~ a. [
b. Limestone, earthy ....................... ; 0
c. Clay .......................................... 0
d. Limestone, irregular masses: 0 to 9
inches ....................................... 0
~- I e. Marl, black and brown .................. 0
~[.f. Limestone, hard, crystalline in the
I
upper portion : thelower simulates
Carboniferous Limestone ............... 0
~g. Marl, hard, shaly .................. seen I)
Thickness in feet inches. Shales, black, clayey ............... seen 2
0
Limestone and quartz-sand
............ 0
4
G~volepis
Alberti;
Oslrea (/)
3
1
5
Gurolepis Alberti. 6
7 Fishes (scales and
10
teeth). In a road-cutting between Merch and Cogan Itall certain Rhmtic
beds are exposed, including a bone-bed :- Thickness in feet inches. 10 to 14 ? Shale, black, clayey : possibly about 4
5
[lepis. 15. Limestone, hard, grey : 0 to 2 inches 0
1 &~hizodus (?) ; Gyro-
16. Shale, black, clayey ..................... 0
6 [" Sauricldh.~/s acumi~a-
17. Arenaeeous rust-coloured deposit ... 0
1 1 tus, Gyrolepis Alber-
18 to 21. Shale, black
............ estimated at 3
0
ti; quartz-pebbles. It is impossible to correlate these beds with certainty, and so
the numbers affixed to them must be regarded merely as suggestive. g
y
gg
About halfway between the foregoing section and the place where
this lane joins the Cogan road, fish-remains are not uncommon in
the Sully Marls. Although these beds are well-developed in this
outlier, it is recorded in the Geological-Survey Memoir that, whereas
a certain zone occurs 26~ feet below the Rh~etie Black Shales at
Lavernock, in this outlier it occurs only 14 feet below that datum-
level)
This may point to some overlap of the Sully Beds (see
p. 413). Vol. 6 ~.]
CO~TIGVOUS DEPOSITS OF GLAMORGANSttIRE.
397 Vol. 6 ~.]
CO~TIGVOUS DEPOSITS OF GLAMORGANSttIRE. 397 397 are found to have changed, both as regards faunal and lithic
characters :~ 1 , The Geology Of the South-Wales Coalfield : Pt. iii--The Geology of tile
Country around Cardiff' Mere. Geol. Surv. 1902, p. 54.
Trans. Cardiff :Nat, Sor vol. xxvii (189t-95) p. 42. 1 , The Geology Of the South-Wales Coalfield : Pt. iii--The Geology of tile
Country around Cardiff' Mere. Geol. Surv. 1902, p. 54. ~,R. T,. R,Cm*RDSO.~ O.X THn R~a~TIC A~D
[Aug. ,905, Newton, who replied :~' This is
the shell which is known as Pallastra arenicola, Strickland.'
Ostrea
llristovi abounds in the Sully Beds at this locality; but, in a deep
wheel-track some 350 yards to the north-east, the equivalent beds Madison on June 20, 2016
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Downloaded from y
, p
Trans. Cardiff :Nat, Sor vol. xxvii (189t-95) p. 42. (A) Coldknap,
Barry. In the low cliff near Coldknap Farm, and facing Barry Island,
is a section of much interest and importance. In the Geological-
Survey Memoir on the Cardiff district it is recorded that ' l~hoetic shalesare exposed again as an inlier near Coldknap. They form a Madison on June 20, 2016
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Downloaded from 398 ~R. L. I~IenARDSO~ O=~ Tnr I~It~TIC X~D
[Aug. I905, i Prec. Geol. Assoc. vol. x (1888) p. 531. 'A small quarry 100 yards south of Redland shows the Avieula.eontorta Shales
overlying a hard yellow dolomitie rock of Tea-Green Marl age, while Carbon-
iferous Limestone, apparently in place, crops out on the opposite side of the
road. Here, then, we can fix a poil~t on the Keuper coast-line, [be the lime-
stone-ground to the north was still above water at the close of the Keuper-
Marl period.' ('Geology of the South-Wales Coalfield : Pt. vi' 1904, p. 30.) , Q
(
) p
" ' The Geology of the South-Wales Coalfield : Pt. iii ' Mem. Geol. Surv.
1902, p. 65, ~R. L. I~IenARDSO~ O=~ Tnr I~It~TIC X~D
[Aug. I905, Madison on June 20, 2016
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Downloaded from 399 CONTI(~IIOUS DEPOSITS OF aLA)IOROAI~ISHIRE. I A. Rendle Short, Quart. dourn. Geol. Soc. vol. lx (1904) p. 171. CONTI(~IIOUS DEPOSITS OF aLA)IOROAI~ISHIRE. The most interesting exposure in this neighbourhood is in the
sides of a field-road, near the brook, about three-fifths of" a mile in
a direction a little to the south of west of Cadoxton Church. I A. Rendle Short, Quart. dourn. Geol. Soc. vol. lx (1904) p. 171.
" ' The Geology of the South-Wales Coalfield : Pt. iii ' Mem. Geol. Surv.
1902, p. 65, ~R. L. I~IenARDSO~ O=~ Tnr I~It~TIC X~D
[Aug. I905, ~R. L. I~IenARDSO~ O=~ Tnr I~It~TIC X~D
[Aug. I905, small anticline running rather south of east, and are traversed by several small
faults, all probably branches from the Coldknap Fault.' A sketch-section is given by Mr. Strahan top. cit. p. 64). Beneath
the Ostrea-limestones are the following beds :~ Thickness in feet inches. Thickness in feet inches. Thickness in feet inches. Paper-Shales. {
( Marl,bluish-grey, with harder bands 5
6 Modiola minima, Moore
Calcite-layer ........................... 0
0~
Band of indurated marl
0
o~_
............ --2
A. ~ Marl, bluish-grey . .................... 0
2
Layer composed of the valves of an
Ostrea ................................. 0
2
t Marl, bluish-grey ..................... 2
10
B. Limestone in two beds, bhfish-grey
weathering yellow. The lower
bed in particular resembles the
Sun-Bed, and exhibits a con-
choi&fl fr,~ctm e
.................. 0
1. Shales, bluish-grey, calcareous, t)
2. Rubbly limestone, with more
compact
limestone
imme-
diately below,
weathering
yellowish. Fossils
most
d
abundant
in
the
upper
I
portion ........................... 0
C.~
3. Shale .............................. 0
4. LunesLone, yellowish
......... 0
5. Shale, blue and yellow, in-J0
'
durated
........................ f
6. Gritty layer
..................... 0
7. Shale, greenish-grey,
thinly
laminated ........................ 0
8. Limestone, blue-centred
...... 0
9. Shale, greenish-grey ... about 0
10. Grit, pyritic, ripple-marked:
0 to 3 inches ..................... 0
UPPER I~ILETIC. 4. Shales, greenish-grey. [non Sowerby. Ostrea sp. 6
5
{ Plicatula intus-striata,
I P1. hettangiensis, and
Lima
(Plagiostoma)
vaioniensis, common ;
5 ~ Protocardium Philip-
I pianum,
PleuroTho-
rus (?), Modiola mini-
~na (?) (large) Moore,
o ~ Ostreasp.; anostracod. " { PleuroThorus (?) & Pro-
i tocardium (?) as casts,
9
1 ~ ~ Plicatula
intus-
slriata. (Lima (Plagiostoma)
9
valoniensis. Shell-ddbris ; Plica/ula
2 ( iMus-striata. 2
3
3 The White-Lias Beds (C) in the foregoing section are extremely
fossiliferous, and it is remarkable that they have not attracted
attention on this account. The strata between the raper-Shales
and the Upper Rhaetie marl or shales (4)at Lavernock measure
8 feet 10~ inches, and at Barry 12 feet 3~ inches, the increase in
thickness at the latter locality amounting therefore to close upon
3 feet 6 inches. Mr. H. B. Woodward has observed a bone-bed in crevices of the
top-bed of marl, at the base of the Black Shales at Cadoxton. l i Prec. Geol. Assoc. vol. x (1888) p. 531. (B) Cadoxton. (B) Cadoxton. Thickness in ,feet i~whes. (B) Cadoxton. Thickness in ,feet i~whes. a. Marls, yellow and black ............... 9 i b. Limestone, hard, dark, in masses
~ I
mixed with dark-brown clay:
"" ]
3 to 9 inches
0
.! m
e. Shales, black, earthy .................. 1
t d. Limestone, hard, dark ......... 0
e. Shales, black and brown, with '3"e]iow
~
streaks. ( Lepidolus (?), Aerodzts
I minimus,
Gyrolepis
Alberti ; bond (Lab.y-
j rinthodon ?) ; Ostrea
6
Bristovi
(see
P1. XXXIII,
~g. 4),
j Pteria (.4vicula) (.:)
8 ~ contorta ; Natica (::)
(i Ostrea Bristovi (rare). The necessity for grouping the Sully Beds with the Rh~etic will
be obvious from the foregoing section, although it should be men-
tioned that the fossils are not individually numerous. Here it will
be noticed that Ostrea Bristovi is associated with _Pteria (Avicula)
contorta and other Rhmtic lamellibranchs. Pteria (Avicula)-contort~t
Black Shales, with
an extremely
fossiliferous Pecten-Bed, are to be seen in the deeply-cut lane three-
fourths of a mile north by east of Cadoxton Church : the limestone-
bed yields Peeten valoniensis, Pteria (Avicula) contorta, Schizodus
Ewaldi, and a Placunopsis similar to that which occurs ill Bed I of
the Rcdland (Bristol) section? Between the last section and Redland (near Bonvilston) lhe
Rhaetic deposits are but seldom exposed. Black shales with thin
Peeten-Limestones have been observed in a brook on the north side
of Bears Wood, south of Wenvoe Castle, and again west of
St. :Nicholas, ' on both sides of the valley and in Coed-y-Cwm.' ~ y
y
In a road-section at, Redland (Sheets 261,262) some interesting
details can be observed. In the Geological-Survey Memoir on the
country around Bridgend is the following passage :~ In August 1904, the section on the west
side of the road exposed
a boss of Carboniferous Limestone wrapped round by Black Shales. This boss, at the road-level, measured 2J feet across, and stood 2 feet
high. A little farther in the direction of Blackland the Black Madison on June 20, 2016
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Downloaded from ~1~. L. mCHARDSO~ ON TrtF, rmmTiC XND
[Aug. 1905, 400 ~1~. L. mCHARDSO~ ON TrtF, rmmTiC XND
[Aug. 'The Geology of the South-Wales Coalfield : Pt. vi--The Geology of the
Country around Bridgend' Mere. Geol. Sur~'. 1~)4, p. 39. (B) Cadoxton. 1905, Shales were more in evidence, and pieces of a Pecten-Limestone
containing the characteristic lamellibranch, fragments of a Placun-
opsis, and fish-scales, were lying about. Still nearer Blaekland
higher beds were exposed, including thin bands of pale limestone
intercalated in similarly-coloured shale, altogether 3 feet 4 inches
thick, and resting upon grey and yellow shale (belonging presumably
to the Upper :Rh~etic), of which a thickness of 6 inches was
visible. Where the a in Redland comes on the 1-inch Geological-
Survey map there is a large quarry in the Carboniferous Limestone. In one place some green marl has been washed down a fissure, but
on the whole, as noticed by :Mr. Cantrill, the Rbtetic soil can be recognized so close to the brow of a large quarry in
Carboniferous Limestone, as to suggest that the Keuper is here wholly over-
lapped, as shown on the map.'l A pond to the east of the ancient camp at Leige Castle is in the
Black Shales, and from pieces of Pecten-Limestone were obtained
Labyriuthodont-bones (small pieces), scales of 6'.~rolepis, coprolites
(fish), and fragments of Pecten (CMa~nys) valo~,iensis. A section
showing the junction of the Rh~etic and Carboniferous-Limestone
deposits can be studied in the roadside 200 yards north-west of
Ty'n-y-coed, but here--as at Redland--the Rha~tic Beds exhibit no
littoral facies, such as might be at first expected. The finest section in the Cowbridge district is in a road-cutting
at Tregyff, near the St. Mary-Church :Road Station on the Cow-
bridge & Aberthaw :Railway. It is dealt with in the Geological-
Survey Memoir (o1~. cir. p. 40) in the following short passage :~ ' They [the Rhmtie deposits] emerge south-east of Wren Castle, and here for
the first time exhibit signs of the oncoming of a more arenaceous type, in the
intercalation of bands of greenish-grey sandstone. These may be traced south-
wards along the valley, but are especially well-shown in a road-cutting at
Tregyff, though the beds undulate gently down the road westwards at such an
angle that only a few feet are exposed. They consist of sandstone with
I'rotocardium Philippiauum and AvicMa contorta, blue sandy marls, a thin
conglomeratic band coutaining pebbles of Carboniferous Limestone and cllert
and black limestone with Peclen valoniensis.' Madison on June 20, 2016
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Downloaded from (C) Tregyff,
near Cowbridge. Thicknes~ in.feet i~whes. i
i Thicknes~ in.feet i~whes. Thicknes~ in.feet i~whes. Limestone, pieces not iu sita. Shales, brown (several t'eet). Limestone, dark ..................... 0
1
Shales, brown ........................ I
3
(
(Shales, black, with several bluish
1
I
gritty layers ........................ 1
i
[ Limest'one, brown and blue ......... 0
~;
[
weathering
......... r ~
] Sandstone,Shale' fine-grained,br~
00
s4 ~
5a~ Shale, black, weathering brown ... 0
6
(i
1 Casts of worm-burrows
7
ton the underside. Madison on June 20, 2016
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Downloaded from 401 Vol. 6x.]
cox'rf(~cous DEPOSITS OF GLA3IORGANSHIRE. d
0
Thickness in
Sandstone, light-grey, fine-grained,
calcareous. Shales, black, thinly-laminated. weathering brown,
and
con-
taining thin gritty layers ......... 1
5b. Limestone, grey,
massive, with
muscovite-flakes
.................. 0
6. Parting, usually of' shale, sometimes
impure limestone .................... (I
7. Limestone, grey, massive ............ 0
8. Shale, black, weathering brown. thinly laminated, with a few gritty
layers ............................ : ....... 0
9. Limestone, grey, thinning out in
places, often arenaeeous ............ tt
~. (Shales,
black,
laminated,
with
t~" ~ several gritty layers near the top,
14'
and at the base, towards the
( eastern end of the section,
a
2
(Massive bed of conglomerate (see
15~ p'402):Ot~
............... i Hard band of grit.: 1 to 3 inches . ~
16. Shales, black, with gritty layers con-
taining a few fish-scales (indeter-
minable) .............................. t}
17. Limestone, hard, grey, slightly sandy
in places, usu-flly in two beds
with a little shaly matter inter-
vening between them ............... (~
IS. Marl, greenish-grey mad bluish," i
imperfectly laminated. ~Near the I
top are little lumps of a hard
t
limestone simulating
Carbon-
iferous Limestone ..................... {'Limestone, hard, compact, greyisb-
I green,
sandy,
resembling,
from
a
di'stanee,
Carbon'
~ I iferous Limestone:
containing
()
i ~ J
occasionally fragments ot'a white ,
earthy
marlstone
and
small
/
s
I quartz-pebbles ........................ )
I Marlstone, yellowish-green ; visible
[ in the gutter ........................ fe...[ inches. 0
3
I)te;'ia
(Avicula)
co,-
torta (.two specimens). "2 ( Pecte~ (Ch!am.fsj va!o-
nie n sis,
Schizodus
lOi
Ewa/,li; Acrodusmini-
i mus, teeth and scales
', of G-yrolepis All~e;'[ i. I
I Pec~en (Chlamys) cal,;-
(i
n iensis,
1)lacunopsis
alpina (large). 4
Much shell-d6bris. I'ech, n (5'hla~,,~y~) calo-
1
niensi.~, Pteria
(Ari-
eula) contorta,coprolite. / Aerodus minimus (com-
mon),
Saurichthys
acuminatus, ' Saryodon
5
to'micas' (Lepidotus ?),
] ichthyodorulite
of
Hybodus;
casts of a
' lamellibt,'aneh of Sehi-
zod~;s-type ; wood? 2
Schizodus ? (casts). (Pleria
(Avic.uga)
con-
J torta ; a few fish-scales
7 5 I (G!/rolcpis ?) and co-
( prolites (fish). (" H?/bodus minor( l toot h),
j
Gyrolepis
Alberti
! (scales, rare); Pteria
4 ~ (Avicula)
contorta,
[ 3Iodiola, fragments of
] lamellibranchs,
and
: carbonaceous matter. / Acrodus ~ninimus (corn-
: mon), ' 8argodo~ to-
ll}{ micas'
(Lepidotus?),
[ fish-scales (fragments
[ common). From
a lithologieal standpoint the Tregyff section presents
man) From a lithologieal standpoint the Tregyff section presents man)-
features of interest. All from the Conglomerate-Bed. Madison on June 20, 2016
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Q.J.G.S.
No. 243. r-
~R. L. nICH.kRDS0.N ON" 'J~ItE RtI~]!~'I~IC A~ND
LAug. I9o 5, grouped with the Sully Beds. The fossils recorded from the lime-
stone-bed were observed mostly iu the uppermost portion. Above
is a deposit of soft marl, which ~'ielded single examples of Pte~'i(~
(Avicula) co~ntorta, Modiola (indeterminable), and ltybo(l~s ~;~ino,'
(tooth). The mos~ interesting stratum in the foregoing section is the
Conglomerate-Bed distinguished as 15. g
g
In a series of deposits such as the Rhmtie, it is often difficult
to correlate with certainty the various component deposits seen
at different localities. In
certain
districts, however,
as in
~'orth-West Gloucestershire and Worcestershire, the same lithic
characters are preserved by a bed over a considerable area; but in
Glamorganshire such is not the rule, and as the beds are traced
westwards there is increasing evidence of the proximity of land. Here, at Trcgyff, the Lower Rh~etic was deposited close to a shore-
line composed of Carboniferous rocks, and as a result the Secondary
rocks becarae what Charles Moore would have termed ' abnormal.'
Therefore, it will be understood that when in the foregoing record,
and in those of sections farther west, a bed is distinguished by a
certain number, that number suggests
rather than implies
contemporaneity with a similarly-notated deposit elsewhere. p
y
y
p
Mr. E. B. Wethered, F.G.S., has kindly supplied me with the
following notes on certain limestone- and chert-pebbles from the
Conglomerate-Bed :~ ' Limestone-pebble from the Conglomerate-Bed, Tregyff, near Cowbridge.--
An impure limestone containing a number of organic remains, but so altered
by molecular change that it is very difficult to determine them beyond a few
spines, crinoid-ossicles, fragments of polyzoa and mollusea. The main feature
of the slide is the presence of a large number of crystals of salt and gypsum,
which are apparently of secondary origin.' 'Chert-pebble from the Conglomerate-Bed, Tregyff, near Cowbridge.--
Looked at in a hand-specimen this appears to be a chert enclosing oolite-
granules. The sections of this chert seen under a microscope show it to
contain a number of ovoid bodies, some of which include a nucleus, but the
usual form of concentric structure characteristic of oolite-granules is not
apparent. Vol. 6x.]
cox'rf(~cous DEPOSITS OF GLA3IORGANSHIRE. At the base are hard marlstoncs which are best 2a 2a Q.J.G.S. No. 243. Q.J.G.S. No. 243. Madison on June 20, 2016
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~R. L. nICH.kRDS0.N ON" 'J~ItE RtI~]!~'I~IC A~ND
LAug. I9o 5, ' Limestone-pebble from the Conglomerate-Bed, Tregyff, near Cowbridge.--
An impure limestone containing a number of organic remains, but so altered
by molecular change that it is very difficult to determine them beyond a few
spines, crinoid-ossicles, fragments of polyzoa and mollusea. The main feature
of the slide is the presence of a large number of crystals of salt and gypsum,
which are apparently of secondary origin.' Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from ' Limestone-pebble from the Conglomerate-Bed, Tregyff, near Cowbridge.--
An impure limestone containing a number of organic remains, but so altered
by molecular change that it is very difficult to determine them beyond a few
spines, crinoid-ossicles, fragments of polyzoa and mollusea. The main feature
of the slide is the presence of a large number of crystals of salt and gypsum,
which are apparently of secondary origin.'
'Chert-pebble from the Conglomerate-Bed, Tregyff, near Cowbridge.--
Looked at in a hand-specimen this appears to be a chert enclosing oolite-
granules.
The sections of this chert seen under a microscope show it to
contain a number of ovoid bodies, some of which include a nucleus, but the
usual form of concentric structure characteristic of oolite-granules is not
apparent.
Originally the rock was probably a limestone, which has been
transformed into chert by the gradual replacement of the carbonate of lhne
by silica.' g
y
"' See also Buekland & Conybeare, Trans. Geol. Soc. see. 2, vol. i (1824)
p. 301 ; and ' The Geology of the South-Wales Coalfield : Pt. vi--The Geology
~)f the Country around Bridgend' Mere. Geol. Surv. 1904, p. 41. 1 A rock crowded with this lamellibranch, but not found in silu, is assigned
to this horizon, owing to the similarity of lithic structure. CoN:rIGUOUS DEPOSITS Ok" GLAMORGAN*SfIIIIE,
403 403 1 A rock crowded with this lamellibranch, but not found in silu, is assigned
to this horizon, owing to the similarity of lithic structure.
"' See also Buekland & Conybeare, Trans. Geol. Soc. see. 2, vol. i (1824)
p. 301 ; and ' The Geology of the South-Wales Coalfield : Pt. vi--The Geology
~)f the Country around Bridgend' Mere. Geol. Surv. 1904, p. 41. r-
~R. L. nICH.kRDS0.N ON" 'J~ItE RtI~]!~'I~IC A~ND
LAug. I9o 5, Originally the rock was probably a limestone, which has been
transformed into chert by the gradual replacement of the carbonate of lhne
by silica.' The Pecten-Beds (7 & 5 b) are well-developed; but the super-
incumbent deposits are difficult to correlate, both on account of the
lack of distinctive fossils and the growth of vegetation, which
obscures the upper portion of the section to a large extent. In the neighbourhood of Ty-ganol the lower portion of the Lower
Rh,-etie consists mainly of u sandstone-deposit. This sandstone is
well exposed by the roadside, 180 yards north-east by north of
Ty-draw, and therefrom were obtained a specimen of H!/bodus rumor
and u few scales of Gy,.olel~is Alberti. The whole of the lower
stage, however, is not replaced by an arenaeeous deposit, because Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from Vol. 6I.] Vol. 6I.] 1 Higher up, in a field on the south side of the road, there is a quarry ill
blue-limestones with shale-partings--the upper beds full of I~iloeeras Joh~tstolzi.
They are crowded with gasteropods, the commonest form of which is Ccrithiu~ll
9ral~t;i~, Terquem. In the quarry o,1 the north side of the road, as noticed by
5ft. I~'. T. Howard, Trans. Cardiff ~Nat. Sac. vol. xxx (1897-98) p. 41, are
several limestone-beds crowded with ~l]lccv.s~i/ia cf. irreyu}aris, Duncan.
Halfway between IAandough-juxta-Cowbridge and Llanfdhangel are two
quarries, one on each side of the valley. In that on the north side is a bed
full of the same species of Theco~mi/ia, while the other fossils included I'lc;,ia
(Avic,ula) i~ceq~tiea/~'i.% Li,,tc( ~,alo~icnsis, Z. g~tbercztluta, ~Iodiola q~zini~na (/),
1)h~a, Or~zithella sarthace~si~, and radioles of an eehinoid. The quarry on
the opposite side yielded P~i!oceras pla~torbis, Lima yiga~lea (small), I~eeten
af~ ealcus, Pi~a, "Ostrea, Modiola ~ti~tima, Sow., and a gasteropod. ' The Geology of the South-~ales Coalfield : I't. ~i' (1904) pp. 41-44.
a Ibid. pp. 44-48 and Trans. Cardiff l~'at. Sac. vol. xxx (1897-98) pp. 36
e~ seqq. ~t~. L. ~,~ClXXl~DSO.',- ox "i'u~: rti~mr~c aZ,,'D
[Aug. ~9o5, Between iIow Mill and The I-[erberts a road-section shows
sandy beds and shale :-- CoN:rIGUOUS DEPOSITS Ok" GLAMORGAN*SfIIIIE BIt. Cantrill has noticed some Pecten-Limestones between Ty-ganol
a,~d Pentre. At Pcntre certain White-Lias beds have been preserved as an
oullier; also a few Rhmtic beds, as proved by pieces of Pecten-
Limestone with fragments of Pecten (Chla;nys)valoniensis. This
outlier is situated 200 yards east of the road, and owes its preserva-
tion to faulting, whereby the beds composing it have been let down
on the north against the Carboniferous Limestone. Formerly
there w,qs a quarry here, but now the excavation is filled with
water. On the south side of this pond the following beds are
exposed :- Thickness in .feet bwhes. f'Clay, yellow, shaly at the base: visible .............................. 1
0
~j| Limestone, brown and dark-coloured; P[iealula inb,s-striata
~
common I ............................................................... 0
5
Clay ........................................................................ 0
3
, Limestone ................................................................. 0
2
Clay, yellow ............................................................... 0
5
Limestone ; 3lodiola ;Mt~ima common .............................. 0
2
(_Shales, dark, hard, with thin beds of limestone ......... seen 1
t; The uppermost limestone in the foregoing record exactly resem-
bles the bed bearing a similarity to the Sun-Bed at Barry (p. 398). The account of these Rh~etic beds given by Mr. Cantrill shows
that, in the St. Hilary district, few if any exposures escaped his
attention. But there are one or two interesting facts not as yet
recorded. By the side of a pond, indicated by the arrow on the
Geological-Survey Map, and some 300 yards to the west of Garn,
is a limestone-bed very much resembl'ing, lithically,
a certain
development of the Estheria-Bed of North-West Gloucestershire. This stratum contains Pseudomonotis fallax, and dips gently to the
north-cast by north: it is, therefore, most probably on the same
horizon as the Pscudomonotis-Bed. In the floor of the lane between St. Hilary and St. Hilary
Common, there is a quite fossiliferous bone-bed. The deposit is a
pale, slightly-arcnaceous limestone, containing obscure casts of a
lamellibranch (Schizodus ?), scales and teeth of Gffrolel)is Albe,'ti,
Ac~'odus minimus, and a few indeterminable fragments of bone. ~ In the roadside at The Cross, south of St. Hilary, the ]lhmtic
sandstones are exposed. There is, however, a considerable amount
of black shaly matter present, :~nd in the sandstone-layers inter-
calated in this shale--especially in that about an inch thick towards
the middle (vertical) of the exposure--'plant-rcmains' are abundant. 2G2 Madison on June 20, 2016
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Downloaded from 404 ,
,
,
,
,
g
p
' The Geology of the South-~ales Coalfield : I't. ~i' (1904) pp. 41-44.
Ibid
44 48
d
C
diff l '
S
l
(189
98) In that part of the memoir on ' The Geology of the South-Wales Coal-field
which deals with the Cardiffdistrict (pt. iii, 190 ~ p. 56) there is the following
passage :--' The fauna includes scarcely any examples of the four great marine
orders, Actinozoa being unknown in the Acietda-coutorta beds, Echinodermata
bein
re resented by one form of feather-~tar Brachiopoda by one form of
g
P
"
9
"''
-"
r
i
~Di~ciua, Cephalopoda by a Bclogcu~hzs, the horizon of whmh, however, the e s
reason to doubt.' Later discoveries require this statement to be modified
somewhat: ia addition to the occurrence of an echinoid in the Ptcria
(Avict,la)-contor/a Beds, a compound coral has been described by the late R. F.
Tomes (Quart. 5ourn. Geol. Soc. vol. lix, 1903, p. 403) ; and ~he same author
tband an imperfectly-preserved :Vonglicallia, parasitic on a Modiola in the Black
Shales, during the construction of the l'ei~arth Docks (ibid. vol. xl, 1SS4,
p. 363).
Charles Moore obtained from his ' Flinty bed' at Beer Crowcombe,
Somerset, a single specimen of a coral, probably a species of 3lol~llivaltic~
(!bid. vol. xvii, 1861, p. 511). Between iIow Mill and The I-[erberts a road-section shows
sandy beds and shale :-- "lhick~tess i~e ./~'ct i,tch~,*. "a. Shale, black ................................. /,. Sandstone, fi~m-grained,
calcareous:
1 to 2 ipehes .............................. 0
c. Shale, grey ................................ 0
d. Limestone, arenaceous, nodular: 1 to
2 inches .................................... 0
e. Shale, black ................................ 0
.~: Sandstone, hard and dark-grey bands
near the base, but olherwise some-
v
what soft .................................... t;
Kr, up~a. I. Hard, grey, rocky marls ............ (_; yro&1)is A//,erti,
and some other fish-
1 remains, bttt indete:'-
( minablel
5
1
1
{
Iu the hard pal
I tions
were
noted
()~ casts of ~t lamelli-
] branch and numerous
small pieces of wood. As no distinctive fossils were obtained from these rocks, it is only
poss.ible to say that thcy belong to the Lower Rhmtic. l The sections in the railway-cuttings north and south of St. Mary-Church Road Station have been described in detail in the
Geological-Survey Memoir on the district';
and, in both cases,
below the representative of the Paper-Shales of the Lavernock
Section, is the White Lias--shaly and marly matter constituting
the greater mass of the deposit. Owing to the ample details
recorded in the memoir just cited, it is unnecessary to discuss
these sections here ; and the same remark applies to the sections of
the Rh,'etie Beds at both ends of the railway-cutting at Cowbridge. :~
That at the southern end is the more satisfactory of the two. Tho
Paper-Shales are 8 inches thick, and the Rhmtie and White-bias
deposits between them and the Keuper measure 19 feet. Having dealt with the sections of the Rhmtic deposits in the
district between Barry and Cowbridge, I may now direct attention Madison on June 20, 2016
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Downloaded from 405 u
6i.] co,tremors Drrosi~s or ~;LA.~0t:G.~.~Sm~. to the outliers in the neighbourhood of Peudoylan, at Peterston,
and St. Fagans2 g
In the Pendoylan outlier, Mr. Cantrill discvvered pieces of
Pecten-Limestone at a pond 100 yards south-east of the vicarage;
and I found fragments of a similar rock at a pond immediately
west of the footpath running from Pendoylan to Ty'n-y-cae, at a
point due west of the Tre-stbch. Mr. Cantrill obtained spines and
tubercles of an echinoid (queried as an AcrosaT.eMa), from a Peeress-
limestone thrown out from a well-excavation, 80 yards north-west
of Pendoylan School. The occurrence of Rh,~tic deposits bas been noted by the officers of the
Geological Survey at certain localities which have not F~en touched upon in
this paper, because, although I ~i~ited these lo:alities, I did not obtaiff any
additional information. The results of the official investigations are chronicled
at the following pages of the memoir on ' The Geology of the South-Wales
Coalfield: Pt. iii'--p. 66 (St. George's); p. 66 (Coed-y-gof); p. 66 and iu
Pt. vi (1904) p. 39 (Castell-y-Mynach) ; p. 65 (Saint welI) ; and p. 65 (Vishwell).
2
Handbook to the Geology of Cheltenham' 1904 p 210 ,
gy
,
Prec. Cotteswold Nat. F.-C. vol. xv (1904) p. 40. Madison on June 20, 2016
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Downloaded from m~. L. RICHARDSON ON THE RII:I,'.TIC AND
[-Aug. I9O5, l{hmtic beds are preserved between two faults, as recorded in the
Geological-Survey Memoir, but they have been omitted~no doubt
inadvertently--from my copy of the map. Between iIow Mill and The I-[erberts a road-section shows
sandy beds and shale :-- The fae~ is interesting, because, except for
the records oi ~ an echinoid (which may be a species of ]'seudodia~lema)
at Coomb Hill near Cheltenham,'-' and at Church Lench (Worcester-
shire), 3 I am not aware that such remains have been noticed in beds
of co~torta-age. ~ Mr. Cantrill, moreover, observed Rh~etie limestone
in the soil of a field, at about 150 yards along the footpath running
south-eastward from Ty'n-y-cae. Near the hedge at the same spot
are shallow excavations, in which fragments of sandstone containing
Acrod~ts misdirects, and bits of fish-scales, can be seen. In the Peterston outlier Black Shales and Pecten-Limcstones have
been noticed in a brook between Maendy and Allt-isaf, bu~ the only
exposure of any interest is at the village of St. Bride's, in a shallow
road-cutting near the church. On the west side a considerable
thickness of black shale occurs, together with a bed of sandstone
crowded with the teeth of Ac;'or
minimums; while on the east
the ' Tea-Green Marls' (Keuper) are visible. There is no evidence
of the Sully Beds. In the sides of a pond about 200 yards west of Pen-hefyd,
St. ]?agans, Black Shales and a PecteJ~-Limestone (containing Pecten
v(doniensis and fragments of a P/acmwj~sis? ) are visible. These Madison on June 20, 2016
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Downloaded from 406 m~. L. RICHARDSON ON THE RII:I,'.TIC AND
[-Aug. I9O5, iii. Cowbridge to Pyle. At most localities between Cowbridge and Pyle the Black Shales
and intercalated sandstone-bands, which usually constitute the deposit
laid down during the contorta-age, are replaced by massive sandstones
containing few fossils; while, durfi~g the time when the Upper
Rhmtic was deposited, a greenish marl with red streaks was formed. p
g
What details concerning the Rha~tie can be obtained in the
neighbourhood of St. Mary ]:Jill have been recorded by Mr. Tidde-
man ~ ; it is, therefore, sufficient to state here that the arenaceous
element predominates over the argillaceous. p
g
The road between the railway near Coychurch and Coity rul~s
along the outcrop of the Rhmtic, of which there are several ex-
posures; as, for instance, seven-tenths of a mile north-west by west
of Coyehurch Church, and again in the road at Simondston. In the
village of Coity itself, sandstones and shales of a greenish tint (but
mottled red in places), and full of ~llodiola ~inim, (?)Sow., are
seen in a road-cutting near the Castle. 1 , Summary of Progress of the Geological Survey for 1899' (1900) p. 129 and
' The Geology o|' the South-Wales Coalfield: l~t. vi--The Geology of tile
Country around Bridgend ' Mere. Geol. Surv. 1904, pp. 50, 51.
2 , Summary of Progress of the Geological Survey for 1899' (1900) p. 129 and
'2"he Geolog3; of the South-Wales Coalfield: :Pt. vi--The Geology of the
Country around :Bridgend' Mere. Geol. Surv. 1904, p. 50.
See also his
pamphlet on the ' Quarella Q,arries,' Bridgeud, 1S93. y
g
, pp
,
2 , Summary of Progress of the Geological Survey for 1899' (1900) p. 129 and
'2"he Geolog3; of the South-Wales Coalfield: :Pt. vi--The Geology of the
Country around :Bridgend' Mere. Geol. Surv. 1904, p. 50.
See also his
pamphlet on the ' Quarella Q,arries,' Bridgeud, 1S93. 1 , Summary of Progress of the Geological Survey for 1899' (1900) p. 129 and
' The Geology o|' the South-Wales Coalfield: l~t. vi--The Geology of tile
Country around Bridgend ' Mere. Geol. Surv. 1904, pp. 50, 51. (B) Quarella
Quarry,
Bridgend. North of Bridgend are the well-known Quarella Qulrries, where
pale-green and white llhmtic sandstones are worked, the rock being
very suitable for building-purposes. A single block from Bed c (of
the sandstone-deposit) of the appended record measured 72 • 53 • 44
inches, and must have weighed about a ton and a half. The deeper
quarry, and that which yields the more satisfactory section, is
situated on the south side of the lane :w Thickness in .]'ec~ iwq~,s. O,rr,~.l- I Limestone, grey. .............................................. 0
t;
Shale-parting ................................................... 0
1
J3EDs. "[ Limestone ...................................................... ()
3
Shales, bluish-grey and brown .............................. 0
4
1. Shales, hard, passing into hard bluish-grey limestones
with couehoidal fracture .................. nmximum 0
7
{
a. Shale, black and brown, clayey . .......................... I)
4
2. b. Shale, hard
................................................. 0
4
C
"7
, e. Marls, greenish and yellowish .............................. 1
,q
3. Limestone, argillaceous, nodular: 3 to 5 inches ...... 0
4
4. Green and yellow sandy marls; ahnost a fine sand-
stone in places .................................... about
6
(;
(a. Sandstone, pale-green and white, rather broken u 1).,. 4
4
5a I I~. S~ndstone ...................................................... 3
4
to ~ e. Sandstone
4
0
15(?) id" Sandstone
4
0
I e. Sandstone, more flaggy, and therefore in thinner layers,
seen S
(; The first notice of this section is contained in Tawney's paper
' On the Western Limit of the Rh,'ctic Beds in South Wales & on the
Positioa of the " Sutton Stone."'
He regarded the sandstone-beds
as belonging to the Keuper, while ' 6 feet of green sandy marls'
he doubtfully referred to the Rhmtic)
Charles Moore observed
(Quart. Journ. Gaol. Soc. vol. xxiii, 1867, p. 513) that ' in the valley west [north ?] of ]3ridgend the Keuper Sandstones are largely
worked, but their succession upwards into the Lies is not well exposed ' ' the sandstones quarried for building and grindsto:ies .... are not situated
at the base of the Keuper as stated by Mr. Tawnev, but are in the upper part
of the Rh,'etic Series, overlain by Lies crowded with the characteristic 0.4,'ca
liassica.' Whichever subdMsions of the Rhwtic are represented here, and
in the absence of fossils (except for a Lima) it is difficult to decide,
it is obvious that we have the equivalents of nearly the whole
series. Vol. 61.]
COlgTIGUOUS DEPOSITS OF GLAMORGXNSttlRE. 407 Q
(
) p
2 , Summary of Progress of the Geological Survey for 1899' (1900) p. 130
and ' The Geology of the South-Wales Coalfield : Pt. vi--The Geology of the
Country around Bridgend' Mere. Geol. Surv. l (!04, p. 51. (A) ttendre,
near Pencoed. ]3etween Hendre and Pencoed the Rh,'ctic has been disturbed by
two faults, that on the north letting it down against clayey shales
belonging to the 3Iitlstone Grit. The Hendre brick-kilns are
situated on the Xeuper Marls. At a slightly-higher level to the
north are the Rh~tic sandstones, faulted against the Millstone Grit. A passage driven from the briekworks to the bottom of the clay-
pit passes throngh the Rhmtic sandstone, and from such fissile rock
numerous ill-defined casts, of a lamellibranch of ,Schizod~s-facies,
were obtained. The Keuper Marls are excellently exposed in a pit
adjoining the works. A little to the east of the tIendre brickworks are two quarries
in which massive Rhmtic sandstone is worked. Pullclstra arenicol~
and scales and fragmentary bones of G!/rolepis Alberti have been
collected in this neighbourhood by Mr. Tiddeman, who has also
given the following record of the seq~lence of deposits 2 :~ 77~iel~'~2ess in feet. LowLa I~n.~:TIC. Sandsto~e, white, fine-grained, massive below and more
thinly-bedded and yellowish abo~e ............... seen 30
{~. Sandy beds ......................................................... 3
UPPnR
[ b. Green and yellow marl .......................................... 15
Kr:t-wa. .{ c. IIard marl, with 'race' and gypsum ........................ 2
[ (1. Clayey red marl ................................................... 2
~e. Clay and marl ; proved in a well ........................... 30 ~e. Clay and marl ; proved in a well .... 1 Quart. Journ. Geol. See. vol xxii (1866) p. 72. Madison on June 20, 2016
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Downloaded from y
g
, p
' See also ' Summary of Progress .f the Geological Survey for 1899' (1900)
p. 131 ; ' The Geology of the South-Wales Coalfield : Ft. vi' Mere. Geol. Surv.
.1~,)04, p. 56 ; and Quart. Journ. Geol. Soe. vo]. xxii (1866) p. 71. ' The Geology of the South-Wales Coalfield : Pt. vi--The Geology of the
Country around Bridgend ' Mere. Geol. Surv. 1904, p. 53. :MR. L. RICHARDSON ON THE l't]~I ".:ETIC AND
[Aug. i9o 5, Sandstone, from which Rhmtic fosdls have been obtained, has
been worked near the Angeltown Asylum, and (more recently) for
building the church at Pen-y-fai. The section in the railway-cutting at Cwrt-Colman is now for the
most part overgrown, but the details recorded by Tawney show
(1) that there is an increase in the proportion of the shale-deposit
to the sandstone as compared with the Quarella section, and (2) the
1)Iesence of a recognizable bone-bed (Quart. Journ. Geol. Sac. vol. xxii, 1866, p. 70). Massive sandstone-beds, however, are
still to be seen oll the south side of the line. Near this cutting,
in a road-section
south of Melin Cwcw, certain information
concerning the upper portion of the Rh,-etic has been obtained
by Mr. Tiddeman. 1 (C) Stormy
Down. The picturesque and breezy moorland known as S t o rm y D o w n
is covered with large masses of Rhmtic sandstone, and is broken with
many scattered openings, most of which have been made in search
of sand for silica-bricks. In a long line of excavations the following
sequence may be observed :~
h
'
f
h Thie~'ne~ in.feet i~zches. ~. Sandstone with an uneven base: 10 to 12 feet seen ........... 11
0
& Marl, green mad yellow, clayey. Soft and hard layers alter-
hate. They appcar to have been slightly contorted previous
to the deposition of the superincumbent sandstone, as ~ft
arenaceous matter fills up the miniature synclines. A little
lignite occurs near the base ................................. abou~
1
10
c. Marl, greyish-green ; full of plant-remains ........................ 1
~
d. Clay, brown ............................................................... 0
2
e. Marl, grey and brown .......................................... seen
(1
~;
[Apparently this deposit extends some feet deeper.]
t: Sandstone of considerable thickness [' Tea-Green ~[arls' ?]. Tawney (o 2. cit. pp. 70, 71) has recorded Pteria (Avicula) comorta
from the maEs intervening between the sandstone-deposits on
Stormy Down ; while from the sandstone (but not in sit~t) I have
obtained casts of Schizodus, MOo phori(~ ,Natica 29yle~sis, Tawney,
and Cylindrites ov~/'ormis, Moore, together with a fragment of the
iehthyodorulite of a Hybodus."
By the roadside near Llangewdd
Court large masses of sandstone were found to contain numerous
plant-remains. (B) Quarella
Quarry,
Bridgend. The stratum distinguished as I much resembles the Cotham-
Marble equivalent; while the nodular limestone (3)is very sugges-
five of the EstheHa-Bed. From the limestones capping tim section
Mr. Tiddeman obtained Ostrea liassica and Pleurom~ta. ~- Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from :MR. L. RICHARDSON ON THE l't]~I ".:ETIC AND
[Aug. i9o 5, '408 (D) Stormy-Down
Cement-Works. At the time of my visit to the Stormy-Down Cement-Works
water had accumulated in the pit, a~d so ]t was only possible to
examine certain of the Lower Liassic beds. The lower beds are,
therefore, tabulated on iMr. H. B. Woodward's authority. Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from ' ' The Jurassic Rocks of Britain' Men,. Geol. Surv. vol. iii (IS93) p 114. co.~I6trolrs DI.;POSlTS OF 6L.~MOR(3ANSl]IItE. specimen of C(,h;cerc, s i~;/.erm,~e!i~:m (Po~-tloek) was fbund,
but not ia ~i[u.] I co.~I6trolrs DI.;POSlTS OF 6L.~MOR(3ANSl]IItE. "l"hid',esr i, f,_'e! bwhcs. (- i. Clay, with yellow and hard grey limestone-nodules;
i
Pe,taerinus- ossicles, l'.,~ud,.Ji,,lema-'..adio!e ? ; Li-m,
(-Pla.qiostoma) yiga,tca, L. (Radv.la) pce~it~oidcs, ~Sti-
cardium cardioidcs, M,,dida minima, 0.4red el. liassicG
I
Pholadomya .qlabra ; De,talium (tah',s'e, gasteropods ;
Psiloccras Johnst,;,i
.................................... s~eu
3
6
2. Limestone ............................................................ 0
;~
4. Limestone, yellowish, nodular: P.'i occr(._~ .J~ ...... s,.,,' ....... l
,S
5. Shale, dark". ...................... ................................. 0
:4
6. Limes!one, bluish, mixed with slm]v matter. ................. 2
7
7. Sh fie dark. u~u~ v o,~i.,tm~t
"
n
l;
9 :
, , ", ~ :1t pv.'~S ...................................... 8. Hard blue limestone, with a little shalv matter; Os~rea
liassica, ISilc, cerasplau,:,rbi~ [seen in 19])5, S feet ; add 10.~ 18
0
. 9. Conglomerate-bed .............................................. 0
II~
'~ 10. Limestone-shales ................................................ 0
8
ttard compact limestone (resembling the Sun-Bed)
...... 0
10
Shaly parting. IIard, compact, and rather shaly limestone .................. IJ
S
Black shales, with thin bands of limestone : l'cc/e~z vcdom-
em.'is ............................................................... 1
0
Grey and greenish marls, with hard nodules (formerly
used for cement) ........................ seen to a depth c~i"
:1
6
[A. specimen of C(,h;cerc, s i~;/.erm,~e!i~:m (Po~-tloek) was fbund,
but not ia ~i[u.] "l"hid',esr i, f,_'e! bwhcs. (- i. Clay, with yellow and hard grey limestone-nodules;
i
Pe,taerinus- ossicles, l'.,~ud,.Ji,,lema-'..adio!e ? ; Li-m,
(-Pla.qiostoma) yiga,tca, L. (Radv.la) pce~it~oidcs, ~Sti-
cardium cardioidcs, M,,dida minima, 0.4red el. liassicG
~ I
Pholadomya .qlabra ; De,talium (tah',s'e, gasteropods ;
Psiloccras Johnst,;,i
.................................... s~eu
3
6
2. Limestone ............................................................ 0
;~
4. Limestone, yellowish, nodular: P.'i occr(._~ .J~ ...... s,.,,' ....... l
,S
5. Shale, dark". ...................... ................................. 0
:4
6. Limes!one, bluish, mixed with slm]v matter. ................. 2
7
7. Sh fie dark. u~u~ v o,~i.,tm~t
"
n
l;
9 :
, , ", ~ :1t pv.'~S ...................................... 8. Hard blue limestone, with a little shalv matter; Os~rea
liassica, ISilc, cerasplau,:,rbi~ [seen in 19])5, S feet ; add 10.~ 18
0
. 9. Conglomerate-bed .............................................. 0
II~
'~ 10. Limestone-shales ................................................ 0
8
ttard compact limestone (resembling the Sun-Bed)
...... 0
10
Shaly parting. IIard, compact, and rather shaly limestone .................. IJ
S
Black shales, with thin bands of limestone : l'cc/e~z vcdom-
em.'is ............................................................... 1
0
Grey and greenish marls, with hard nodules (formerly
used for cement) ........................ seen to a depth c~i"
:1
6
[A. Vol. 6 t.]
co.~I6trolrs DI.;POSlTS OF 6L.~MOR(3ANSl]IItE. 4_00 Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from air,. I~. ~lCn.~r, DSO:~ ox 'rm: R~a~IC A~'D [Aug. 19o5, There is obviously some peculiarity about the stratigraphical
sequence in this section. Observers arc a~reed that at the 1sase
there are greenish marls: Tomes mentioned them as belonging to
the Keuper ; Mr. H. B. Woodward as belonging to the Rhtetie. It'
they belong to the latter, then they must correspond either to the
green marls above the sandstone at the Quarella Quarry (p. 407), or
to some bed occupying the stratigraphical position of that which is
lettered b in the section on Storm) Down (p. 4'J$), if it be considered
that they correspond to the former, the presence of ' black shales, with
thin bands of limestone ; Pectr v(donie~sis,' in the foregoing section
at Stormy-Down Cement-Works (p. 409), needs explanation ; and
again, if to the latter, the absence of the sandstone-deposit, such as
that seen on Stormy Down (Bed (,) requires accounting for. Charles
Moore, as already mentioned, commented upon the insignificance
here of the e(luivalcnt of the Rhmtic of the West of England. Tomes thought that the greenish marls belonged to the Keuper, and
if his surmise be correct the Rh,'etic Black Sh.ales probably rest
thereupon non-sequentially, for the Peeress-Beds occur at some height
above the base of the series, where the se~luence is complete. p
Not having had the opportunity of examining the beds which
may possibly correspond to those that compose the greater portion of
the Upper Rh,'ctie at other localities, I think that it is undesirable
to offer any suggestions. The Rh~etic Beds in the neighbourhood of Pyle are largely
represented by sandstones. In the recently-published Geological-
Survey Memoir on the district, there is an excellent account of the
sections available, and to that account I have nothing to add. ~
Attention, however, may be directed to the fact that Tawney appears
to have studied beds higher in the series than any seen of late years,
because he makes mention of limestones ' which .... from their
appearance and conchoidal fracture, remind one of the Cotham
Marble . . .'~
He prefaced his observations on this tract with the
remark that' The above-mentioned patch [was] sufficiently described
by Mr. Bristow,' and referred the reader to Rep. Brit. Assoc. 1864
(Jhath) Trans. Sections, p. 50. air,. I~. ~lCn.~r, DSO:~ ox 'rm: R~a~IC A~'D [Aug. 19o5, In the place cited there is no mention
made of this Pyle, but Pvlle Hill (Bristol) is referred to as a
locality visited by Bristow, for the purpose of making a detailed
section of the Rh~ctic Series. In addition to Mr. It. B. Woo(lward,' several other authors have
described this section,
tI. W. Bristow wrote : In addition to Mr. It. B. Woo(lward,' several other authors have
described this section,
tI. W. Bristow wrote : 'The section a~ the Stormy Cement-\Vorks . . . shows about 20 feet of
ordinary Lias limestone and shale, resting upon 2 feet of a hard, siliceous, and
shelly blue conglomerate, under which occur from 12 to 15 inches of p:fle
argillaceous limestones, breaking with a smooth eonehoidal fi.acture, and which
I believe to represent the " White Lids" or uppermost, member of the I~ha?tie
Series.'
(Quart. Journ. Geol. See. vol. xxiii, 1867, p. 204.) Charles ~oore, in his valuable eontribntion
to our knowledge of
the Mesozoic littoral deposits, commented upon t.he fact tha~ 'When compared with the West,-of-England section, the lChwtie beds at this
spot are very insignificant. A single bed of black marl containing Pec~e;~
c~donie~2.sis and other Rh,~tic shells succeeds the variegated marls, aud upon
this a dark limestone 4 inches, and next a bed (in texture very similar to the
" White Lias") 2 feet thick. T'he Os&ea-beds then follow .... ' (Il:id. p. 520.) In 1.884 the late It.. F. Tomes recorded in the pages of the
Quarterly Journal (vol. xl, p. 359) the following details of the stra',a
below the Ostrea-Beds :-- Tl~i.k, ess i~ .~;'t i,ch(.~. ' Hard conglomerate, in all respects like the " Guinea"-Bed of
Binton ............................................................... 2 to
:3
0
Fine-grained nodu!ar limestone, with a eonchoidal fracture, and
very much resembling the l'2sthe~'ia-Bed ..................... 1 to
2
0
Dark-~ey R.hmtic shale ................................................... 1
6
Greenish, compact Keuper Marl, forming the bottom of the pit.' Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from 410 1 , The Geology of the South-~Va]es Coalfield : Ft. vi~The Geology of the
Country around Bridgcud ' Mere. Geol. Surv. 1904, pp. 5t-55. 1 , The Geology of the South-~Va]es Coalfield : Ft. vi~The Geology of the
Country around Bridgcud ' Mere. Geol. Surv. 1904, pp. 5t-55.
" Quart. Journ. Geol. Soc. vo]. xxii (1866)1). 70. y
g
" Quart. Journ. Geol. Soc. vo]. xxii (1866)1). 70. IV. ADDI'I:IONAL OBSERVATIO.NS ON TIIE SULLY BEDS. It is geuerally admitted that, towards the close of Keuper times,
there was a great inland sea covering a large part of England,
which by degrees evaporated. As Mr. A. Rcndle Short has pointed
out, the conditions were probably desertic, and therefore over that
area there would be a more or less uniformly-horizontal surface,
wil:h gently-shelving shores, and occasional deeper pools and Madison on June 20, 2016
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Downloaded from 411 Vol. 6I.J Vol. 6I.J co~Iovovs DEeOSlrS or GLX3m~a.~Smr, E. channels. ~ This inland sea, at the close of the epoch, had bee~l
reduced by evaporation to a few comparatively-shallow lakes sur-
rounded by fiats of mark y
One of these lakes certainly existed in the Lavernock district,
and, as I have already pointed out, it would be in such areas that
transition-beds between the Keuper and the l(hmtic should be
sought for. 2
This lake was deepened, and its limits further
restricted, by earth-pressures which occurred at the close of the
Keuper Epoch. p
p
It is difficult, at present, to decide whether the earth-pressures
were more intense at the commencement or at the close of thc time
when the Sully Beds were deposited. y
p
It is well-known that towards, or at the close of, the Carbon-
i~'erous Period, earth-pressures affected the Palmozoic rocks of
most parts of the world, and among them Glamorganshire, and
caused the strata to be thrown into a number of anticlines a~d
synclines. The folds in Glamorganshire have been mapped and
admirably described by Messrs. Strahan & Cantrill, the mat,
axis being distinguished
by these authors as the Cardiff-
Cowbridge anticline. As observed by Mr. Strahan, this anti-
cline as far west as Pendovlan was simply a broad arch, but in that
neighbourhood became compound. Mr. Cantrill has shown the
position of the subsidiary anticlinal fold: it trends a little north
of east and south of west, and brings to the surface at Stalling
])own, near Cowbridge, quartzitic and pebbly beds belonging to the
Old Red Sandstone. The movements which occurred at the close of the Keuper Epoch. and affected the newly-formed conglomerates and marls, wer(~
probably of upheaval along old anticlinal axes, although of cours(. they need not, and it would appear did not, agree with these pre-
cisely in position in every case. ' Quart. Journ. Geol. Soe. vol. lx (1904) p. 186.
2 It, id. p. 357. Madison on June 20, 2016
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Downloaded from I The debated question as to the age of the Sutton Stone is now a thing of
the past: ' the whole of these beds may be regarded as the basement-beds
of the Lower Lias, representing the O~'~'ca-13eds and other portions of the zone
of Ammo~ziles planorbis, and including perhaps portions of the zone of A. a~gl.-
latus' (H. B. Woodward, ' The Geology of the South-Wales Coalfield: Pt. vi'
]~[em. Geol. Surv. 1904, p. 62). I concur with this view. A little to the west
of the Caves, below West, near Southerndown, a quarry (now disused) had
been opened out in beds about the junction of the Sutton Collglomm"ate and
Stone, and from the spoil-heap were collected Chemuitzia (fragment), Anemia :
(with Serpula), A~4arle ( Cardita ? rhomboidalis, Tawney), Cardima regularly,
Terq. (=C. su#o~m~zsis, Towney), Gruph~ea, Lima (Cle~wstreon) tuberculate,
Terq., Lima (Playiostoma) sp. (small), Lima (Radula) he/ta~tgiensis, Terq.
(juv. = L. subdtq)licata, Tawney), Odrea ~aul/icostala, Miinster, Plicatula intu~-
striata, :Emmrieh, 1u
caloMensis, Defrance, MJilus imbrieato-radiMa,
Tawney, LithoThagus sp., Serpula (~), and Astrocomia gibbosa (Duncan). Lima
Terquemi is not uncommon, and this fossil alone demonstrates the non-Rha~tic
and the non-~,Vhite Lias nature of the deposit in which it ~curs. IV. ADDI'I:IONAL OBSERVATIO.NS ON TIIE SULLY BEDS. One of the main axes of elevatiol~
at this time, howevcr, was apparently along the line of country
traversed by the subsidiary pre-Triassie anticline referred to above. On the hillside to the south of Aberthin, near Cowbridge, rock
thought to be Lower Lias is shown in the Geological-Survey Map
to rest directly upon the Keuper; and bctween this outlier and
Cowbridge there is another outlier of the same rock, which is depicted
as resting upon the littoral Keuper. The Rhmtic has been sho~vn
by the railway-cuttings at Cowbridge to intervene at those particular
localities between the littoral modifications of Keuper aad Lower
Lias; but usually in the Cowbridge outlier the Lower Lias rests
directly, and therefore non-sequentially, upon the Keuper. Similar
phenomena are observed at Llanbleiddian--a village near Cowbridge. Now, if deposition had continued unchecked from Keuper to Liassic
times, then everywhere the sequence would have been--Keuper. l~h~etic, and Lias; we should not have seen Lias resting directly
upon Keuper. Only two explanations can be suggested: either
the Rh,'etic has been deposited and subsequently removed (but in Now, if deposition had continued unchecked from Keuper to Liassic
times, then everywhere the sequence would have been--Keuper. l~h~etic, and Lias; we should not have seen Lias resting directly
upon Keuper. Only two explanations can be suggested: either
the Rh,'etic has been deposited and subsequently removed (but in Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from =
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i I
.............. :,,
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0 Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from 4i;~ u
6i.] u
6i.] 'r)aE l~n.mlc Dl~voslis ov G~.t.~ml~(;.~xsniI:~. pre-Liassic times), or the Rha~tic has not been deposited. mR. L. RICHARDSON ON TIlE IlH:ETIC AND
[Aug. I90~, Keuper. No mention is made in the Geological-Survey publications
of the abundant occurrence of Ostrea in the 'Tea-Green Marls' at
St. ~[ary's-Well Bay, Sully : it is observed that these marls 'are distinguished from the Rhmtic formation by the fact that they have
hitherto proved to be totally unfossiliferous in South Wales, except for the
occurrence of a bone-bed at Goldcliff. 't This statement, however, is modified in a subsequent part of the
same memoir, 2 but the same classification is adhered to. All, hough
I have beell compelled, from the pal,'eontological evidence, to suggt,st
that the Sully Beds be classed with the Rhmtic, I nevertheless quite
agree that the line of demarcation between Keuper and Rh~etic
decided upon by the officers of the Geological Survey is the mo~t
satisfactory for their purpose, because it is then possible to indicate
on the map where the lithic change takes placeJ 1)~enewed earth-movements occurred before the deposition of the
Rhmtic Black Shales, and brought about conditions suitable for a
slight erosion of the uppermost stratum of the Sully Beds. In the
I)enarth-Lavernock section, indications of such an erosion are most
apparent. pp
I would here direct attention to the fact that Mr. iF. T. Howard,
F.G.S., is of opinion that ' in South Wales, at least, there was a slight upheaval instead of a sub-
sidenee just at the close of the Trias Period (immediately preceding the
subsidence of :Rhretic and Liassie times, about whieh the evidence is indis-
putable). We see at once that the newly-formed Trias beds might be raised
i~to dry land, and immediately attacked by the various agents of denudation.' * Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from 414 mR. L. RICHARDSON ON TIlE IlH:ETIC AND
[Aug. I90~, p
p
2 Ibid. 2t. iii--" The Country around Cardiff' 1902, p. 41.
a Since this paper was written, that dealing with the Rhmtie rocks of
Monmouthshire has been communicated to tile Geological Society.
In the
discussion which ensued (Quart. Journ. Geol. See. vol. lxi, 1905, p. ;384)
5Jr. Strahan said that he had been unable to identify the 'Grey Marls' of
Etheridge, and doubted whether any such subdivision of the' Tea-Green Marls'
~ould be made. Tile pal~eontological evidence set forth in the present paper
requires such a division ; but, of course, the actual line of demarcation between
the Sully Beds and the ' Tea-Green Marls' must be more or less arbitrary, told
governecl solely by palmontoiogical considerations. Mr. 8trahan also doubted
whether there was any o~erlapping of the ' Tea-Green Marls' by the Avicula-
conlorta Shales, and remarked that, they varied but little in thickness over
the whole region.
According to the Geological-Survey publications, the
' Tea-Green Marls' at Lis-Werry measure 13 feet (' Geology of the South
Wales Coalfield: Pt. i--The Country around Newport, Men.' Mere. Geol.
Surv. 1899, p. 74) and at Lavernoek 4a.89 feet (ibid. Pt. iii~' The Country
around Cardiff' 1902, p. 54)--a difference of 31~ feet. , p
)
Trans. Cardiff Nat. Soc. vol. xxix (1896-97) pp. 65-66, ' The Geology of the South Wales Coalfield : Pt. i--The Country around
Newport, Men.' Mem. Geol. Surv. 1899, p. 70. IV. ADDI'I:IONAL OBSERVATIO.NS ON TIIE SULLY BEDS. In either
case, it seems to me lhat it is necessary to invoke the assistance of
e:~rth-movemm,ts to explain matters; but, taking all the facts
available into account, it would certainly seem that at the localities
in question the ]~h,'etie had not been deposited, owing to an elevation
which was initiated about the time of the deposition of the Sully
Beds. On the westerly continuation of the Cardiff-Cowbridge anti-
cline at Bevos, near Tythegston, it is interesting to notice that the. relations of the Lias to the Keuper also suggest that the above is
the correct interpretation of the phenomena; while at Sutton again
it is very doubtful whether the Keuper Conglomerate was ever
parted by a Rhmtic deposit fi'om the Sutton Conglomerate.' p
y
p
g
In the stretch of water which extended into the Lavernock
district, and the outlines of which had been modified by the earth-
pressures referred to above, the Sully Beds were formed, while the
'Tea-Green Marls' at certain other localities were undergoing
suba~rial denudation. When the Rh,'etic ocean gained access to
the British region it spread over the Lavernock area; its waters
mingled with those of the Lavernoek lake; the sediment too was
commingled, and the resultant deposit has the peculiar lithic
structure which characterizes Etheridge's 'Grey Marls' and the
upper portion now denominated the Sully Beds. The appended
diagram (fig. 3, p. 412) will help to ehtcidate my views as regards
the relationship of the Pteria (Aviculu)-co~to,'ta Black Shales to
the subjacent marls and Sully Beds. Throughout the period during
which the Sully Beds were in process of formation, earth-pressures
may have affected the rocks, and it is especially desirable that
exact records should be kept of any sections showing the junction
of these beds wi~h the overlying Black Shales, in order to see
whether there is definite evidence of any overlap of the upper
strata of the Sully Beds on to the ' Tea-Green Marls.' y
It appears imperative that these Sully Beds should be grouped
with the Rhmtie, and consequently a certain portion of the ' Tea-
Green Marls,' as defined during the re-survey of the district
(1892-1901), must be assigned to that series and removed from the Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from coxxlal:OVS I)EPOSITS OF GLA3IOIIGANSI[IRI'. In the neighbourhood of Bridgend, a considerable deposit of sand-
stone was made while black sediment was being laid down around
Lavernock. The very different nature of these contemporaneous
deposits is to be accounted for, mainly by the fact that there was
only imperfect connection between the two areas of deposition:
land, composed of Palmozoie reeks, intervened. p
There is evidence (which will be quoted shortly), however, to
show that near the commencement of the Upper Rh,'etic ' Age ' the
deposits in the Lavernock area were elevated, and a stretch of water
separated-off to the north. Theoretically, this stretch of water may
be regarded as having been connected with that in which the
Upper Rh~etic beds now visible at Bishton, near Newport, ~ were
deposited. This view explains the similarity noticed between the
beds distinguished as 3 at the Quarella Quarry, Bridgend, and
Bishton, and likewise between the remaining similarly-notated
deposits. p
The effects of these earth-movements were not merely local; by
the elevation of certain areas, shallow lagoons were formed in
others, initiating conditions suitable for the existence of Estheria. While the Upper Rh~etie Beds 3 to 1 were in the course of
formation, the Lavernock area was above sea-level ; the marl-deposit
(4) was subjected to subaSrial denudation and somewhat fissured
(fig. 4, p. 412). 1 Quart. Journ. Geol. See. vol. lxi (1905) pp. 377-78. V. CONDITIONS OF DEPOSITION OF THE I{H::ETIC BLACK StlALES. V. CONDITIONS OF DEPOSITION OF THE I{H::ETIC BLACK StlALES. So far as is known at present, while the Black Shales or sand-
stones containing Pteria (Avic,la) co~to~'ta were being deposited in the
Glamorganshire district, no appreciable earth-movements occurred. Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from 415 Vol. 6I.] Vol. 6I.] coxxlal:OVS I)EPOSITS OF GLA3IOIIGANSI[IRI'. ~UR. L. ~,lCItAPrSOX o.'," ~nn m~'l'Ie A.~D
[Aug. 19o5, 1)l. hett(t~*gi~',*sis, and Lir~za valonie~,sis being especially abundant. A stratum which probably represents the Sun-Bed completes the
equivalent of the White Lias of the Bath district : the representa-
tive of Bed A of the Lavernock and Barry sections is only found over
a restricted area, and probably indicates a deepening of the se~t
in this neighbourhood, caused by renewed earth-pressures. 1)l. hett(t~*gi~',*sis, and Lir~za valonie~,sis being especially abundant. A stratum which probably represents the Sun-Bed completes the
equivalent of the White Lias of the Bath district : the representa-
tive of Bed A of the Lavernock and Barry sections is only found over
a restricted area, and probably indicates a deepening of the se~t
in this neighbourhood, caused by renewed earth-pressures. g
,
y
p
During the period when the White-Lias limestones and shales
(A & B, Lavernoek) were being deposited, the Cotham Marble in
certain localities was hardened, broken up, and cemented into a
conglomerate~for example, at Sedbury Cliff', near Chepstow. As
remarked by Dr. Arthur Vaughan, ' The time occupied by the hardeni~g of the Cotham layer [at Sedbury Cliff'2,
its destruction, and subsequent cementation into a conglomerate may be ec,n-
sidered to correspond roughly to the ti,ne cf deposition of the White Lias in
the areas on the south and cast.'
(Quart. gourn. Geol. 8oe. vol. lix, 1903,
p. 397.) It is interesting to observe also that the same author thought
that ' At Sedbury Cliff the deposition of the Cotham Marble must have been
succeeded by an elevation of the floor, which produced the breaking-up of the
Cotham-Marble layer in si[~:.' (Ibid. p. 398.) At the time when the Lower Liassie ' I'aper-Shalcs' were formed,
the bathymetric conditions were about the same in the West of
:England. In the description of the various sections in this shire, reference
has been made to the stratigraphical position of the White Lias, and
it will have been noticed that this deposit is considered to come
above the Cotham Marble. Recent work accomplished by several
geologists in the Bristol district has demonstrated that such is its
correct stratigraphical pcsition. g
p
p
As long ago as 1864, Bristow & Etheridge, in their account or'
the Rh~etic beds at Penarth, referred to ' bands of limestone and
indurated marl (in brown shale),' which they considered might bc
'equivalent to the " White Lias." ... VI. O~ T~E WHITE Li,s OF GLAMORGASSHIRE, AND ON ]:lie
STRATI6I',APItICAL POSlTIOX OF TIlE WIIlTE Llxs. VI. O~ T~E WHITE Li,s OF GLAMORGASSHIRE, AND ON ]:lie
STRATI6I',APItICAL POSlTIOX OF TIlE WIIlTE Llxs. A gradual subsidence in the Lavernoek area, accompanied by the
reverse movement to the north, allowed again of deposition, aT,(1
into the fissured marls (4)gritty nmterial was washed. The
relations of the White Lias and the Upper Rhmtic Beds may be
represented as shown in fig. 5, below. s.w. White /
Lies
Fig. 5.--Diagrammatic section showing the relationship of the
lVhite Lies to the Upper Rhcetic in the Cardiff dls6"ict. Lavernock
near Cardiff
A
B--
C
4
Gol dclfff
near Newport
__ji
Ostrea-beds and Paper-s?aa'les
~-2 "l Upper
~
'
~
t
Rhaetic
.-lr,icula-conlor[a shales
In the Bath district, the sea in which the well-known White
Lias was deposited was deeper than at Lavernoek. At the latter
locality, the various lamellibranehs are usually fcund grouped
upon the surfaces of the limestone-bands: Plic,mda intus-st,'iata,
1 Quart. Journ. Geol. See. vol. lxi (1905) pp. 377-78. Fig. 5.--Diagrammatic section showing the relationship of the
lVhite Lies to the Upper Rhcetic in the Cardiff dls6"ict. Fig. 5.--Diagrammatic section showing the relationship of the
lVhite Lies to the Upper Rhcetic in the Cardiff dls6"ict. Ostrea-beds and Paper-s?aa'les
~
' .-lr,icula-conlor[a shales .-lr,icula-conlor[a shales In the Bath district, the sea in which the well-known White
Lias was deposited was deeper than at Lavernoek. At the latter
locality, the various lamellibranehs are usually fcund grouped
upon the surfaces of the limestone-bands: Plic,mda intus-st,'iata, In the Bath district, the sea in which the well-known White
Lias was deposited was deeper than at Lavernoek. At the latter
locality, the various lamellibranehs are usually fcund grouped
upon the surfaces of the limestone-bands: Plic,mda intus-st,'iata, In the Bath district, the sea in which the well-known White
Lias was deposited was deeper than at Lavernoek. At the latter
locality, the various lamellibranehs are usually fcund grouped
upon the surfaces of the limestone-bands: Plic,mda intus-st,'iata, 1 Quart. Journ. Geol. See. vol. lxi (1905) pp. 377-78. Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from 416 ~UR. L. ~,lCItAPrSOX o.'," ~nn m~'l'Ie A.~D
[Aug. 19o5, u
6i.]
COI~TI6UOIrS DEPOSITS OF OL.~)tOR6A~SnH~E. At Wainlode Cliff, on the banks of the Severn between Gloucester
and Tewkesbury, no White Lias is present:
the Paper-Shales rest
directly upon the Pso~do,~onotis-Bed, as shown in the following
section :~ 7hie]."ne~,~ in feet inche~. O.,rm~a-]]i.:l~s. Limestone, hard, blue ............ 0
4
Osl,'ea lic~iea, Mo:li-
Paper-Shales, brown aud grey, finely
laminated,
with
an
intermittent
limestone-band ........................... 1
1. Limestone. P~cudomo~w:L~-Bed. ' In-
sect-Limestone.'
Hard,
dark-grey
,
and blue ................................... 0
~5
"
"2. Shales, blue and brown, laminated,
weathering into a marly clay ......... 5
=
3. Limestone. E~H~evia-Bed. ' ('r
'-
,. Bed. Hard, yellow, nodular; arbor-
L~
escent markings; irregular fracture... 0
4. Shales, pate, greenish-yellow, coarsely
laminated, marly ........................ 6
LOWEIt } Pte;ia (Aeieula).eomol"ta Zone. 1[{ II.ET tO,
oh~ minima. (" P~eudomon otis
rid_
[ lax,aDa~win ula lias-
3 ~ den, D. liassiea vat. I raffler (teste Brodio
i & Prof. T. R. Jones). ( 3led iola
rain ira a,
5 ~ Ostrea(testeBrodie);
( insects; fish-scale. ( Lyeopodites
laneeo-
2 I lalu,; L~.theria rai-
l m~.la var. Brodieana;
o~traeoda
(teste
I Brodie);
fish- re-
6
mains
aud
shell-
( d(~bris. ( 3lodiola
mini,m,. 0 ~ Schizodu,~ Ewaldi,
~. C~,rdium ~.l,)aeinum. ,
~5
"
=
L~ ~ear
Chase .Hill, Wickwar, the thin flaggy limestones of the
Ostrea-I~eds rest directly upon the Cotham Marble, 2 and in that
neighbourhood the bed frequently contains Pseudo~nonotis fallax in
abundance. But ~o the south, in the railway-cutting at Lilliput,
near Chipping Sodbury, the sequence is as follows :-- Thickness i,~
O,s'rRL~-B~.I~S. Thin flaggy limeslones, separated
by thin shales .................. 3
YV,rTE LIAS. :I3. Compact cream - coloured
limestone ..................... 1
1. Cothana Marble ........................ 0
r5
['Grey shale, containing plant-beds
~. [ at 15 and 18 inches from the top. 2
[ Brown, unfossiliferous shale ...... 1
~"
2 ] Arzillaceous limestone ............... 0
;~
3 ~ Brown or grey shale, with lenti-
4
cular beds and co,,cretions of
argillaceous limestone at two or
t
three levels ........................ 5
LowE,',
R,m~:TIe. 3 Pteria (Avicub,)-conlo~'ta Zone. feet inches. { Ostrea liassica, Mo-
O
diola minima, and
Pleuromy:,. 0
5
3 Damt~,int~la & Es:heria. 0
6
[ Fragmentary shells,
0
Cardium cloacinum,
~ Modiola. Ill the foregoing section, the position of the White Lias is clearly
shown:
it occurs
above
the Cotham
Marble. 1 P~eudomonoti,, de~.~,ssa[, of my previous papers.
2 Geol. Mag. 1904, p. 5:$4. ~UR. L. ~,lCItAPrSOX o.'," ~nn m~'l'Ie A.~D
[Aug. 19o5, The place of the Cotham
Marble (not observed here),' they added, 'is at the base of this
group of beds.'
(Geol. Mag. vol. i, p. 237.) g
p
g
p
It has been remarked recently that the denomination White
Lias
is a useful stratigraphical term for the Upper Rhmtie
Beds in the region extending southwards from Bath (Geol. Mag. 1905, p. 79). Since the term Upper l(ha~tic has been applied by different
authors to obviously non-contemporaneous strata, it is desirable to
define exactly to what beds it has been applied in tt;e present paper. The 'Upper ~htetic' of my records of the sections in :North-West
Glouceste~shire, and those at Wood :Norton (near Evesham), Sr
Cliff (near Chepstow), and elsewhere, is n o t the equivalent of the
White Lias. Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from 417 ,,
,
[,
2 Geol. Mag. 1904, p. 5:$4. .~ln. I~, ~:ICF[~.~DSO~ * O~ TIlE lt]L'~rl'IC A~D
~A~Ugo ~905,
Lodge, near Siston, the White-Lias beds exposed are 418 as follows :~
2hh'k#ess in feet inches. ( O~irea liassicq, 15"oto-
[Limestones, thin, with clay-partings. ! t Clay
0
Limestone, in two beds with clay-parting. 0
J Clay, full of Oslrea : ._z to 1~ inches ... 0
b:
Limestone, sometimes ill two beds ...... 0
(Clay or limestone ........................ 0
I jLimestone, often inconspicuous ...... 0
kBrown and dark clay : :3 to 6 inches. 0
(B. Sun-Bed ................................... 1
~1
Clay ....................................... 0
.. ]
Limestone, thin ........................... 0
Limestone ................................ 0
:.= ! Clay-parting ............................ I)
~[
Limestone ................................ 0
Shaly deposit ........................... 0
C. :Rubbly beds ......................... seen 1
[COTHA.~I M.~RBr, E.]
cm'dium
_Philip-
~, pianum. 2
3}
Pho&domya ylabra. 1
O.stre(, [i(,ssica. { Protoeardium Ph ilip-
4
Ticmum common. 1
1
4
1
2
7
O~ { Out,'ea liassic(,, Lima
~ [ valonie~sis, 31,diola
~ninima,
Pro:,ear-
1 J dimn I)hiZil>l>iro~,m,
! Plicagula
intus-
~,. eh'iala. [COTHA.~I M.~RBr, E.] The Sun-Bed is exposed in the quarry, but it will be observed
that below it is a deposit of which a thickness of 2 feet 2 inches is
seen, without any evidence of the Cotham Marble. But no doubt that
stratum would be found, it" the excavation were a little deeper; and
then, below again, would occur the grey marls of the Upper Rhmtie,
such as those seen in close proximity to the Carboniferous Limestone
seven-tenths of a mile south by 5 ~ east of Abson Church) The rubbly beds (C in the foregoing section) are interesting. They seem to be present, as a rule, in the Bath district, but are
rarely exposed as they are of no industrial value, and quarrying
operations are therefore not prosecuted so deep down. These beds
are not, present: in the Lilliput section, but I recently (May 1905)
obtained evidence, from d~bris thrown out of a well, to show that
they do extend farther north than Blue Lodge, Siston, where they At an ochre-working, at the cross-roads near Gathcram Farm, Wick
Rocks, the following interesting section is exposed :-- Thick~zess in feet i~ehes. {'Light-coloured, grey and yellow clay
I
seen
6
:Peculiar deposit of black and brown
sandy clay,
impure limestone
i
with
in places .................................... iN t Sandy bed, with more or less angular
I
pieces of grit (the biggest measuring
~. 2xl-~xl inch) : 89 to 2 inches ......... At an ochre-working, at the cross-roads near Gathcram Farm, Wick
Rocks, the following interesting section is exposed :-- u
6i.]
COI~TI6UOIrS DEPOSITS OF OL.~)tOR6A~SnH~E. Some 5 miles south-south-west of Lilliput the White Lias is seen
to have increased in thickness ; for, in a quarry by the side of the _'~ vt _'~ vt Q. J. G. c.~ ~o. 24"r Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from co~'rlGcocs DEPOSII:S OF GLXMORGA~SHIRE. co~'rlGcocs DEPOSII:S OF GLXMORGA~SHIRE. are seen at the bottom of the quarry noticed above (p. 418). Unfortunately, this well, which is situated in the orchard at the back
of the inn in the village of Codringtou, had been sunk some weeks
previous to mv visit to this part of the country, and the greater mass
of the rock wlaich had been excavated had been carted away. y
The well is 74) feet deep, and the water-supply good. Those
blocks of Lower Lias which were sufficiently compact had been
used for walling the sides of the well, so that very little of the rock
was lying about. The limestone which could be examined con-
tained small specimens of Lima yiy, ntea and Ostrea liassh'a. How
great a thickness of Lower Lias was penetrated could not be ascer-
tained. Pieces of a compact cream-coloured limestone showed that
the White-Lias Sun-Bed had been proved, together with the sub-
jacent, rubbly, fossiliferous White Lias, which contained Plicatula
hettangiensis (one specimen),
Lima
vcdo~de;~sis, Protoca,'dit*~;t
Philil~piaaum , Modiola mbdma, Moore, and gasteropoda. Masses
of a hard limestone (with a mammillated surface and faint arbor-
escent markings)showed that the Cotham Marble was present, if
not in its quite
typical form. The remaining Upper Rhmtie
deposits are apl)arently similar to those which were exposed in the
railway-cutting at Lilliput, and some of the indurated portions con-
tained fish-scales and teeth not uncommonly. Black shales were
very much ia evidence, and contained a hard dark limestone with
Pecten ( Chlamys) valoniensis, Pteria ( At,ic~r
co,torta, Plact,.~Ol)Sis
a!pina (large), and ,~c!~iz,)du~ (1) : also dark, calcareous, very mica-
ceous, and occasionally pyrit'c sandstone-layers. A number of
pieces of Bone-Bed were found. They indicated that the Bone-
Bed here was not of the same lithic structure as that at Lilliput,
doubtless owing to the fact that it does not rest upon any member
of the Carboniferons System or the Old Red Sandstone. This is
known from arenaeeous ' Tea-Green Marl' having been found
among the ddbris. The Bone-Bed is a dark-grey, micaceous,
sandy limestone, with Gyrolepis Alberti (.scales and teeth ?), Acrodus
minimus, and coprolites of fishes ; but it, passes into a thin pyritic
sandstone containing a few vertebrate-remains. The excavation
was terminated in the Carboniferous Limestone. .~ln. I~, ~:ICF[~.~DSO~ * O~ TIlE lt]L'~rl'IC A~D
~A~Ugo ~905,
Lodge, near Siston, the White-Lias beds exposed are 2
9
=~a~ (Light-yellow and grey clay, with hard
7~ ~ ]
masses of marlstone containing small
~ t
quartz-pebbles ..................... about 2
6
.= ,~
Reddish deposit, with yellow streaks... 1
6
:~ g
Yellow stone ................................. 1
0
~
I Reddish stone, with vcllow streaks ...... 1
3
B~[~ Hard, yellow stone : ~5 to 7 inches ...... 0
6
~ I Working for ochre ........................ 0
3
:' Sarf/odo~
in'miens
(L@idotus ?), Ac,'o-
0
1 ~ dus :minimus (rare),
; Gyrolepis Albergi, &
fragments of bone. Madison on June 20, 2016
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Downloaded from Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from 419 Vol. 6~.] Vol. 6~.] 1 The White Lias is exposed ill road-sections at :Barrow Hill, aboul a mile
north of the Barton-Farm Quarry; while the junction of the l:tha~tic and
Keuper is seen in the road-eutti~g a short distance to the west, or some
300 yards east of Wick Forge.
• thin deFosit of red sand, containing Acrodu.s
mini mus,
SauHcldh.qs ac~Mnalu.~,
G~'olqd~ Albcrti (scales), 'Sa~:qodo~l
to~nic~ts' (Zepid,lus :t), fragments of bones, and small quartz-pebbles, prob'lbly
represents the Bone-Bed.
o Proe. Bath _Nat. lIi-t. & An'iq. F.-C. vol. ii (1S71-7;3) pp. 204-11. :nr:. r,. mcnxr~DSO.X o~," TttE mt_m'ic .,t~'D [Aug. I9o 5, As I have indicated above, the rubbly beds of the Wldte Lias
probably occur not far below the floor of the quarry, and rest upon
the Cotham Marble. The strata below the Sun-Bed and above the
rubbly deposits at Blue Lodge are 2 feet 2 inches thick (see p. 418):
here, however, the equivalent deposit must be of double that
thickness, since White-Lias limestones, 4 feet thick, are exposed
without any indication of the rubbly beds) y
y
)
Some 4 miles distant from the Barton-Farm Quarry is the instruc-
tive section at Newbridge Hill, on the outskirts of Bath. When I
visited the section, the Cotham Marble was the lowest bed that
could be studied irt sire. The following record is compiled from
details noted by the Rev. H. H. Winwood. F.G.S.," my own notes
being in square'brackets :~ "lhh,/,.,c~ i, ./~c! i,~chc~,.. O
9
G "lhh,/,.,c~ i, ./~c! i,~chc~,.. ]3. Sun-]3ed ........................... O
9~ Gasteropoda; Lima,
Limestones, with an occasional
I'edc~z,
Iqicat~da
parting of clay .................. 5
1 :~,
iMus-driata,Modioh~
C. Rubbly limestone with clay-}
[afl:miMma, Moore],
"~-IIITE i
partings, passing downward t
3lgaciles
[Plcnro. LIas. t
into bluish bands. Very fossi- ] 5
0
m~/ee], AxMus, Cm'-
]
liferous towards the base
... J
d[icm rheticum [fro-
I
Blue clay . .......................... 0
l ~-
loca;'dium
Phililg-
l
Limestone ........................... 0
4
[,iaml,t], [Os/.rc~,7. ~,
lleddish-brown clay ............... 0
6.~
([1] Landseal,)e Stone [or Cotham
Acicuh,
[I'seJo-
[
Marblej ........................... 0
t;~tmo,i.oli~fullax"
UPPER ! J-'2~ Light-blue or grey shales ...... [ l':~the;,ir~ ;,dnula and
R,,.,:T,('.[ I5 j Darker-shaded
band
about t 3
8 .' traces of ,egetabh,
the centre
..................... j
[ n,atter in the band
[4] Light-blue or grey shales ...... about the centre. From these few sections it will be observed, (1) that the White
Lias comes above
the Cotham Marble and below
the Ostrea-
Beds; and (2) that it increases in thickness, as it is traced from
north to south. The component beds of the White Lias successively
overlap one another, and this implies elevation
in the north,
depression
in the south:
a[conclusion also arrived at from
investigations in Glamorganshire. co~'rlGcocs DEPOSII:S OF GLXMORGA~SHIRE. Just 2 miles in a direction a little east of south of Blue Lodge the
White Lias is quarried near Barton ]:'arm, Upton Cheney, the
following section being exposed :~ 2"hic]~'~e~ in fi.et inehe~. Glay ....................................... 0
0
Limestone, rubbly ..................... 0
3 / Osh'ea lia..'~ic.,
Clay ....................................... 0
3i lqeuromya crow-
Limestone: four beds, with clay
OSTliEA-J between the middle two... 0
7 "~ combeie,, Tate non
BEDS. ]
......... I Moore, Protoca,'-
[ dium I'hiHppicmum. partings .............................. 1
0
[Clay ....................................... 0
4
~hlvr~ lB. Sun-Bed, capping hard cream-
LIAS. ~
coloured limestones ............... 4
0
f
White-Lias rubbly beds.]
Cothanl M~rble.]
"211 Madison on June 20, 2016
at University of Wisconsin -
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Downloaded from 420 :nr:. r,. mcnxr~DSO.X o~," TttE mt_m'ic .,t~'D [Aug. I9o 5, p
o Proe. Bath _Nat. lIi-t. & An'iq. F.-C. vol. ii (1S71-7;3) pp. 204-11. COXTI6L'OtS DI;rOSl'lS 0" 6LI.XIOR~X-IIIIU.:. (it) Descriptions are given of sections, the most important of
which are at Lavernock (near Cardiff), Barry, Tregyff (near
Cowbridge), Quarella (Bridgend), and Stormy Down. (it) Descriptions are given of sections, the most important of
which are at Lavernock (near Cardiff), Barry, Tregyff (near
Cowbridge), Quarella (Bridgend), and Stormy Down. g ), Q
(
g
),
y
(iii) Earth-pressures affected the rocks during the formation of
the Sully Beds. The Pteria (Avic~da)-contorta Black Shales rest
with perfect parallelism, but, nevertheless, non-sequentially, upon
the Sully Beds. g ), Q
(
g
),
y
(iii) Earth-pressures affected the rocks during the formation of
the Sully Beds. The Pteria (Avic~da)-contorta Black Shales rest
with perfect parallelism, but, nevertheless, non-sequentially, upon
the Sully Beds. y
(iv) Owing to a local upheaval of the Lavernoek district, early
in the age during which the Upper ]lhmtic Stage was deposited,
only a portion of the lowest bed of that stage is found, and this
deposit was subjected to suba(;rial denudation during the accumu-
lation of the remaining Upper Rha~tic beds elsewhere. y
(iv) Owing to a local upheaval of the Lavernoek district, early
in the age during which the Upper ]lhmtic Stage was deposited,
only a portion of the lowest bed of that stage is found, and this
deposit was subjected to suba(;rial denudation during the accumu-
lation of the remaining Upper Rha~tic beds elsewhere. g
pp
(v) Subsidence in the Lavernock district allowed of the deposition
of the White Lias. As a result, the White Lias rests non-sequentially
upon a portion of the lowest Upper 12hattie deposit. p
p
pp
p
(vi) The White Lias at certain localities in Glamorganshire
contains in abundance P/icat,da i~t~rs-striata, P1. hetta,~gie~sis, and
Likuta valonie~sis. The deposit intervening between the Sun-Bed
and the Upper ]lhwtic near Bath (Newbridge Hill) is over 11 feet
thick: at Lavcrnock the equivalent deposit ,neasures but 2 feet
288 inches. At the last-named locality, above the probable equi-
valent of the Sun-Bed, are marls 6 feet 4 inches thick, which are
grouped with the White Lias. Above come the Lower Liassie
Paper-Shales, succeeded by the Ostrea-Beds. p
,
y
(vii) The Upper l{hmtic of North-West Glouccstershire and
Worcestershire is n ot the equivalent of the White Lias of the Bath
district: the White Lias comes above the Cotham Marble (the
topmost bed of the Upper Rh,'etic) and below the Paper-Shales,
which occur immediately below the Ostrea-Beds. VII. Sc3i~iti'. (i) The Sully Beds, or the upper portion of Etheridge's 'Grey
Marls,' belong to the Rhmtic Series, as shown by the fossils, and
are distinct flom the ' Tea-Green Marls ' of the Keupcr, which do
not contain fossils. Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from Vol. 6I.] Vol. 6I.] 421 COXTI6L'OtS DI;rOSl'lS 0" 6LI.XIOR~X-IIIIU.:. Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from OSTR~X BmsTovr, Etberidge, MS.
(P1. XXXIlI, fig. 4.) Ir
valve.--The dimensions of a~L average-sized specimen
are : length 45 millimetres, breadth 55 ram. The greatest length
seems to be attained (as a rule) at 23 ram. from the most extended
portion of the ventral margin. The valve is flat or slightly convex ;
the ventral margin regularly convex ; while the anterior and pos-
terior margins converge regularly in the direction of the beak,
which unfortunately is not preserved in the specimens collected. The test is somewhat thin for an Ostrea, and composed of numerous
considerably-imbricating layers. Unfortunately, the preservation of the specimens of this lamelli-
branch does not admit of a very exact diagnosis. Mr. E. T. Newton,
F.R.S., informed me (i~t lift.) that specimens of this oyster are
preserved in the Museum of Practical Geology, Jermyn Street,
London, and bear the MS. name of Ostrea Br~stovi, Etheridge. ,
,
g
Like most of its tribe, this form is diflieult to describe ; but, on
account of its frequent occurrence in the Lavernock area, it seems
desirable to have some name wherewith to record it when obtained. The specimen figured is from the upper portion of the Sully Beds
(ghmtic) at Cadoxton (Glamorganshire). C.~Rolr~ cr.oAclsr.~, Quenstedt. (Pl. XXXIII, fig. 5.) C.~Rolr~ cr.oAclsr.~, Quenstedt. (Pl. XXXIII, fig. 5.) g
,p
p
,
g
1856. Oppel, A., & Suess, E., ' Ueber die muthmasslichcn .Equiwdente der K(;sst, um"
Sehiehten in Sehw;d)cn ' Sitzungsber. k. Akad. \Visscnseh. Wien, vol. xxi,
p. 510 & 1)1. if, fig. 2. h
h llh
i
d
f h
h
h
i
i p
)
, g
1901. V'mghan, A., "The llhwtic Beds of the South-Wah's I)irect Line' Quart. Journ. Geol. Soc. vol. lx, Pl). 207-208 & fig. 6. In the immaiure forms of this shell the costm are rounded or
subaeute, and increase in breadth towards the ventral margin ;
while the intercostal spaces exceed the breadth of the eostw. In
the adult forms, however, the costw are usually broader and flat-
topped, while the intercostal spaces are reduced to mere linear
grooves. g
The first definite record of this shell was at Wainlode Cliff~;
but since then it has been found at many localities, and was
especially abundant at Lilliput, near Chipp'ing Sodbury, on the
South-Wales Direct Line. Dr. Vaughan has given an excellent
~.escription of the species, elucidated by figures in the text (Quart. Journ. Geol. See. vol. Ix, pp. COXTI6L'OtS DI;rOSl'lS 0" 6LI.XIOR~X-IIIIU.:. I am much indebted to a number of geologists and others
for kind assistance. For the trouble which he took ill procuring
photographs of the Lavernock sections I have to record a debt of
gratitude to Mr. John Storrie (son of the late John Storrie), of Cardiff;
while, ill obtaining the literature on the Rh~etic of the county I
have received much help fi-om Mr. H. B. Woodward, F.R.S., Mr. F. T. Howard, F.GS., Mr. J. Storrie, and Mr. G. H. Dutton, F.G.S. Valuable assistance with regard to some of the fossils has been most
courteously accorded to me by the Rev. J. F. Blake, F.G.S., Mr. S. S. Buckman, F.G.S., Mr. E. T. Newton, F.R.S., Mr. A. C. Seward, F.R.S.,
and Dr. A. Vaughan, F.G.S. ; while to _~[r. E. B. Wethered, F.G.S.,
and Dr. C. G. Cullis, b'.G.S., I acknowledge my indebtedness for
help in certain petrological matters. For assistance in the field-
work I tender my best thanks to Mr. E. Talbot Paris ; while I feel
that any expression of thanks inadequately conveys my gratitude
to Mr. J. W. Tutcher for preparing the excellent photographs of
ibssils from which the figures ir~ Pl. XXXIII were reproduced. Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from 422 1 P~'(:e. C,ttesw. Nit. F.-C. vol. xiv (1903) table i, facing p. 174. VIII. PAL.~.0_XXO~0GmL ~-OTES. OSTR~X BmsTovr, Etberidge, MS. (P1. XXXIlI, fig. 4.) OSTR~X BmsTovr, Etberidge, MS. (P1. XXXIlI, fig. 4.) OSTR~X BmsTovr, Etberidge, MS.
(P1. XXXIlI, fig. 4.) 20T-2()8), and has drawn attention to
the Cardita-like aspect of the shell, which is especially noticeable
in the specimens figured in the present paper. Charles Moore,
indeed, referred this shell (which he obtained fi'om the railway-
cutting at Willsbridge, near Bitten) to the genus Cardita, and
entered it as such in his section (Quart. Journ. Geol. See. vol. xsiii,
1867, p. 498). His specimen is now in the Bath Museum, and
came from the Upper llh~etie. Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from 423 1 See Quart. Journ. Geol. See. eel. lxi (1[05) p. 378. CONTI6UOUS DEPOSITS OF ~LA)tOI~6.~NStlIRE. CONTI6UOUS DEPOSITS OF ~LA)tOI~6.~NStlIRE. The specimen figured in P1. XXX[II
was obtained from Bed
5b (3), Lower Rh,'etic. Section by the roadside near Bishton,
near Newport (.~[onmouthshire). 1 Pr, IC.~'rrnA IN~us-srnlA'r.~, Emmrich. (P1. XXX[II, fig. 1.) 1853. 'Geogno~tische Beohaehtuugen arts den i;stlichen t)ayerischen & dell augrenz-
enden 5sterreichi~chen Alpen ' Jahrb. der k.-k. geol. lieichsanst, eel. iv, p. 376. enden 5sterreichi chen Alpen Jahrb. der k. k. geol. lieichsanst, eel. iv, p. 376. 1853. Hauer, F. Ritter win, ' Ueber die Gliederung der Trias-, Lias- und Juragebilde
in den nordi;~tliehen All)eu' Jahrb. der k.-k. geol. Eeichsanst. eel. iv, p. 738. ;
)
g
, p
186~. ])umortier, E., ' l~tu(les pal6ontologiques sur h,s l)6pbts jurassiques du Bas~in
du Rh6ue : pt. i--Infralias' Pl). 7g-77 & pl. i, figs. 13-16. p
)
g
p
,
g
1861. Ost~'ea interstriata. Moore, C., 'On the Zones of the Lower Lias & the
Avieula.eoMo~.ta Z(,m, ' Quart. Journ. Geol. Soc. eel. xvii, p. 501 & pl. xvi,
fig. 05. This lamellibranch is too well-known to require any additional
description, but it has not been recorded previously from the Penarth
or Barry districts ; only from beds of later date to the west, as at
Bridgend and near Sutton. At Lavernock it is fifirly common, while
at Barry it is very abundant in the White Lias. I have not recorded
it fi'om the Lower llh,-etic in Glamorganshire; but that it doc:~
occur abundantly in that stage at certain localities (Watcher, for
example) I have been able to satisfy myself only recently (June
1905). At Blue Anchor, near Watcher, it is associated with Pteria
(Avic~da) co,tortq. P~IC~ULA m~'rT~'(;IEXSlS, Terquem. (P1. XXXIII, fig. 2.) P~IC~ULA m~'rT~'(;IEXSlS, Terquem.
(P1. XXXIII, fig. 2.) gaughan lbr Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from 424 'rrlr lllLETIC BEP0SlTS OF t~LAMOtlGANStIIRE [Aug. I9o 5, 'rrlr lllLETIC BEP0SlTS OF t~LAMOtlGANStIIRE. [Aug. I9o 5, comparison with those of the same genus from ' the main Pecte~,-
horizon ' at Lilliput, near Chipping Sodbury, ~ was returned with the
remarks' typical clavellate form.' P~IC~ULA m~'rT~'(;IEXSlS, Terquem.
(P1. XXXIII, fig. 2.) 1855. ' Paldontologie dc l'l~tage infdrieur de la Formation liasique de la Province (it. Luxembourg, (h'and-1)ueh6 (Hollande), et de Hettauge' Mdm. See. G6ol. France, set. "2. ~ol. v, 1). 326 & pl. xxiv. figs. 3-L ,
, )
p
g
186~. Renevier, E., ' Notice~ gdol. & pald,)nt, sur le~ Alpes vaudoises : I--' Infralia~
& Zom. a .~ri('Ma c,mtorta (Etage llhwtien) ' I)P-3'3.-39 & l;l. iii, fig. 4. 1861,. I)mnortier, E., ' Etudes paldontologiques sur les Ddl)6ts jurassiques du Bassin
(lu Rh6uc : i)t. i--Infralias' pp. 73-74 & pl. xii, figs. 4-7, 10. ,
, )
p
g
186~. Renevier, E., ' Notice~ gdol. & pald,)nt, sur le~ Alpes vaudoises : I--' Infralia~
& Zom. a .~ri('Ma c,mtorta (Etage llhwtien) ' I)P-3'3.-39 & l;l. iii, fig. 4. 1861,. I)mnortier, E., ' Etudes paldontologiques sur les Ddl)6ts jurassiques du Bassin
(lu Rh6uc : i)t. i--Infralias' pp. 73-74 & pl. xii, figs. 4-7, 10. Plicat~da hetta,,gieJ~sis has not been recorded previously, to my
knowledge, from the White Lias of this county. It is very common
at Coldknap, Barry, and Lavernoek, and it is difficul~ to understand
how this Plicatv.la has managed to escape detection for so long a
time. It is quite possible, however, that it has been recorded
under another name (Ost~.ea multicostata ?). A specimen submitted
for examination to the Rev. J. F. Blake was at, once identified with
Terquem's fossil. q
As remarked by Tcrquem (ol). cir. p. 326), and later by Prof. Renevicr (02). tit. p. 77), this shell presents several varieties in form. One specimen from Barry agreed precisely with the figure given
by the latter author. LI~.t v.~iomE_~sIs (Dcfl'ance). (P1. XXXIII, fig. 3.) LI~.t v.~iomE_~sIs (Dcfl'ance). (P1. XXXIII, fig. 3.) 1825. 'Mdlnoire gd'ologique sur quelques Terrains de la Normandie oceidentale
pat" M. de Caumont, Mdm. See. Linn. du Cah'ados [postea de Norlnandie
eel. ii, p. 507 & Atlas, pl. xxii, fig. 7. 1825. 'Mdlnoire gd'ologique sur quelques Terrains de la Normandie oceidentale
pat" M. de Caumont, Mdm. See. Linn. du Cah'ados [postea de Norlnandie
eel. ii, p. 507 & Atlas, pl. xxii, fig. 7. This fossil is of frequent occurrence in the White Lias at Laver-
neck and Barry. .k specimen submitted to Dr. A. 1 Quart. ffourn. Geol. Soe. vol. lx ~1904") p. 202. [~For the Discussion, see p. 430..] Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from EXPLANATION OF PLATE XXXIII. [Tile fossils figured in this plate are preserved in the Author's collection.] Fig. 1. PlicatMa iMus-slriata, Emmrieh. X 2. White Lias ; Coldknap, Barry (Glamorganshire). 2. PlicatMa l~elta~.qiensis, Terquem. Natural size. White Lias ; Coldknap, Barry (Glamorganshire). 2. PlicatMa l~elta~.qiensis, Terquem. Natural size. White Lias ; Coldknap, Barry (Glamorganshire). 3. Limct valoMc~sis, Defranee. X about 189
White Lias ; Coldknap, Barry (Gl~unorgan 3. Limct valoMc~sis, Defranee. X about 189
White Lias ; Coldknap, Barry (Gl~unorgans 4. Oslrea }~rislori, Etheridge, MS. Natural size. Sully Beds (Lower tih~e;ie) ; St. Mary's-Well Bay,
shire). 5. Cardium c[oacimlm, Quenstedt. Natural size. Bed 5 b (Lower Rhmtic) ; roadside section, four-fifths of a mile north-
north-east of Bishlon Chureh, near INewport (SIonmouthshire). 1 Quart. ffourn. Geol. Soe. vol. lx ~1904") p. 202. [~For the Discussion, see p. 430..] Madison on June 20, 2016
at University of Wisconsin -
http://jgslegacy.lyellcollection.org/
Downloaded from ON Tt[I~. per ends of the corallites
een noticed, are slightly
se views of the corallites
d the walls between the
narrow, and are insepa-
ome instances the united
r septa, and are even less
e ; in others the thickness 102
P
The caliees and t
the sides and wall
worn down. The
thus given are mo
lumina of neighbo
rable, being perfect
walls are not thicke
than one tenth of t
is greater, but at t
hi d
f h
di 02 d recall the parts of some
y quickly. The columella is
circular in exact transverse
n, and in some places almost
tout. The longer septa are
ze of the corallite, and there
The smaller septa are placed
ar inwards. section of tile colony made at
urface, presents many scores
others more or less obliquely
orallites are wonderfully pre-
ith the manner in which the
been rendered apparent by
es, and in the varieties of the
ctions of the different indi-
erse section is very different
the colony. The portions
ransverse and not oblique may be eight, nine, o
between the larger, an
The surface of a pol
the depth of an inch
of corallites, some cut
to their long axes. T
served, and one canno
ancient crowding of
the variation in the ca
polygonal outlines ret
viduals (fig. 3). The shape of the c q
y
g
y
g
q
g
occurs in lines, and the other forms evidently depend upon ir
ressure during upward growth. EXPLANATION OF PLATE XXXIII. Very deformed corallite
elate to budding individuals and to those which have just
ndependent of the parent. The diameter of the corallites varies from 1"25 mm. to 2.2
nd these measurements refer to corallites which have the
eptal number; one half of the dense tissue between the
f one corallite and that of its neighbour is included in th
urement. This one half represents the proper mural tissu
orallite. The thickness of the wall of the corallites me
bove is one eighth of a millimetre and one twelfth of a mill
hus the structure intermediate between the inside or ca
wo neighbouring corallites is one quarter or one sixth of
metre wide. :In some par~s, however, the width of the unite
ay be nearly or quite equal to that of the calibre of a c
igs. 4, 5). ccurs in lines, and the other forms evidently depend upon ir
ressure during upward growth. Very deformed corallite
elate to budding individuals and to those which have just
ndependent of the parent. The diameter of the corallites varies from 1"25 mm. to 2.2
nd these measurements refer to corallites which have the
eptal number; one half of the dense tissue between the
f one corallite and that of its neighbour is included in th
urement. This one half represents the proper mural tiss
orallite. The thickness of the wall of the corallites m
bove is one eighth of a millimetre and one twelfth of a mill
hus the structure intermediate between the inside or ca
wo neighbouring corallites is one quarter or one sixth of
metre wide. :In some par~s, however, the width of the unite
ay be nearly or quite equal to that of the calibre of a c
gs. 4, 5).
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https://openalex.org/W2968032323
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Open Science
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Cellular MicroRNA Expression Profile of Chicken Macrophages Infected with Newcastle Disease Virus Vaccine Strain LaSota
|
Jiaqi Mu
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English
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Spoken
| 6,490
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Received: 27 June 2019; Accepted: 6 August 2019; Published: 9 August 2019 Abstract: Vaccines with live, low-virulence Newcastle disease virus (NDV) strains are still the most
accepted prevention and control strategies for combating Newcastle disease (ND), a major viral
disease that hampers the development of the poultry industry worldwide. However, the mechanism
underlying vaccine-mediated innate cell immune responses remains unclear. Here, a high-throughput
Illumina sequencing approach was employed to determine cellular miRNA expression profiles in
chicken macrophages infected with the LaSota virus, a widely used vaccine strain for mass vaccination
programs against ND in poultry. Compared to the control group, 112 and 115 differentially
expressed (DE) miRNAs were identified at 24 hpi (hours post inoculation) and 48 hpi, respectively. Meanwhile, 174 DE miRNAs were identified between 24 hpi and 48 hpi. Furthermore, 12 upregulated
and 6 downregulated DE miRNAs were observed in common at 24 and 48 hpi compared with
0 hpi. In addition, target prediction and functional analysis of these DE miRNAs revealed
significant enrichment for several signaling pathways, especially in the immune-related genes
and pathways, such as the RIG-I-like receptor signaling pathway, NOD-like receptor signaling
pathway, and mitogen-activated protein kinase (MAPK) signaling pathway. Our findings not only
lay the foundations for further investigating the roles and regulatory mechanisms of miRNA in
vaccine-mediated innate cellular immune responses, but also extend new insights into the interactions
between the host and NDV infection. Keywords: Newcastle disease virus; vaccine strain; chicken macrophages; microRNA; LaSot pathogens pathogens pathogens pathogens Brief Report Brief Report
Cellular MicroRNA Expression Profile of Chicken
Macrophages Infected with Newcastle Disease Virus
Vaccine Strain LaSota Jiaqi Mu 1,†, Xinxin Liu 2,†, Xibing Yu 1,†, Junjiao Li 1, Yidong Fei 1, Zhuang Ding 1,* and
Renfu Yin 1,* 1
Department of Veterinary Preventive Medicine, College of Veterinary Medicine, Jilin University,
Xi’an Road 5333, Changchun 130062, China
2
College of Food Science and Engineering, Jilin University, Xi’an Road 5333, Changchun 130062, China
*
Correspondence: dingzhuang@jlu.edu.cn (Z.D.); yin@jlu.edu.cn (R.Y.)
†
These authors contributed equally to this work. www.mdpi.com/journal/pathogens Pathogens 2019, 8, 123; doi:10.3390/pathogens8030123 1. Introduction Newcastle disease (ND), caused by virulent Newcastle disease virus (NDV), an Orthoavulavirus,
is one of the most devastating diseases that affects the poultry industry worldwide [1]. ND is listed
in the World Organization for Animal Health (also named Office International des Epizooties, OIE)
Terrestrial Animal Health Code as an economically significant pathogen for avian species and products,
and any isolate from poultry infected with mesogenic and velogenic strains must be reported to the
OIE [2]. NDV is a non-segmented, negative, single-stranded RNA virus with a 15 kb genome that
contains six genes encoding nucleoprotein (NP), phosphoprotein (P), matrix protein (M), fusion protein
(F), haemagglutinin–neuraminidase (HN), polymerase (L), and two additional non-structural proteins
(V and W, that are expressed by RNA editing of the P gene) [3,4]. Depending on the severity of the
clinical manifestations observed in chickens and the tropism of the virus, NDV has been classified into
the following five pathotypes: Asymptomatic enteric, lentogenic, mesogenic, viscerotropic velogenic, Pathogens 2019, 8, 123; doi:10.3390/pathogens8030123 www.mdpi.com/journal/pathogens Pathogens 2019, 8, 123 2 of 14 and neurotropic velogenic [5,6]. Velogenic NDV has high virulence, mesogenic NDV has medium
virulence, and lentogenic strains have low virulence or are non-virulent. and neurotropic velogenic [5,6]. Velogenic NDV has high virulence, mesogenic NDV has medium
virulence, and lentogenic strains have low virulence or are non-virulent. Vaccines with inactivated oil emulsion of the same viruses and live, low-virulence viruses are
the most accepted prevention and control strategies for combating ND in different countries and
regions around the world [6]. As is widely known, NDV infects via intranasal, oral, and ocular
routes and elicits robust mucosal, cell-mediated, and humoral immune responses [7]. The well-known
natural low-virulent NDV strain LaSota, which is a class II, genotype II virus [8], has been the most
widely used live NDV vaccine in chickens for the last six decades and has a good stability and
safety record [9]. However, the specific mechanism of the live, low-virulent vaccine strain LaSota
in vaccine-mediated cellular immune responses from the sides of the innate immune cells, such as
macrophages, remains unclear. p
g
Chicken macrophages, like their mammalian counterpart, as one of the first lines of defense
against microbial infection, play essential roles in both innate and adaptive immune responses [10–14]. Chicken macrophages are also considered to be one of the main target cells for NDV infection and
growth in vivo [15]. 1. Introduction Therefore, additional research is required to explore the functional roles of
chicken macrophages in NDV vaccinations to help understand the mechanistic details of immune
responses during viral infections and the ability of NDV to infect macrophages. miRNA are important
post-transcriptional regulators that are widely expressed in both animal and plant species. Numerous
miRNAs have been identified as being involved in the host–pathogen interaction and regulating the
expression of immune genes and pathways [16]. However, the cellular miRNA profiles in chicken
macrophages in response to the NDV vaccine strain LaSota infection have not been investigated. In the
current study, we examined the cellular miRNA profiles in chicken macrophages in order to increase
our understanding of innate cell immune responses and provide miRNA functional analysis for the
immune-related pathway of the NDV vaccine strain LaSota infection. Therefore, the data presented
herein have provided new viewpoints into the interactions between the host and NDV infection. 2.1. Preliminary Analysis of Small RNA Sequencing To investigate the effects of LaSota strain infection at different time points on a cellular miRNA
expression profile, three small-RNA libraries from HD11 cells infected with NDV for either 24 h post
inoculation (hpi), 48 hpi, or 0 hpi were constructed and then sequenced using a deep sequencing
method. In total, 53,096,137, 58,073,878, and 67,815,716 filtered high-quality reads were obtained from
cells of 24 hpi, 48 hpi, and 0 hpi, respectively. Meanwhile, the majority of the small RNAs ranged
between 21–23 nt in length in the three libraries; the most abundant size class was 22 nt (40.48–44%),
which was consistent with the typical size range of miRNA (Figure 1A). In addition, the clean reads
were aligned to repeat-associated RNAs to find matched reads and were annotated with rRNA, scRNA,
snRNA, snoRNA, and tRNA from Rfam (Figure 1B). Therefore, 29,293,525 counts (39.54%) at 0 hpi,
25,662,627 counts (44.57%) at 24 hpi, and 24,021,010 counts (37.89%) at 48 hpi were annotated with
mature miRNAs, respectively (Figure 1B). Among these counts, a total of 1401 mature miRNAs
(including 615 known and 786 novel), 1361 mature miRNAs (including 605 known and 756 novel),
and 1380 mature miRNAs (including 612 known and 768 novel) were identified at 24 hpi, 48 hpi, and
0 hpi, respectively. 3 of 14 3 Pathogens 2019, 8, 123
Pathogens 2019, 8, x FO Figure 1. Overview of the deep sequencing length data and cellular miRNA differential expression profile in chicken macrophages infected with the LaSota strain. (
Length distribution of total sRNAs in LaSota-infected at different time points and mock infected HD11 cells; (B) analyses of the small RNA reads generated by de
sequencing of 0 hpi, 24 hpi, and 48 hpi small RNA library. Figure 1. Overview of the deep sequencing length data and cellular miRNA differential expression profile in chicken macrophages infected with the LaSota strain. (A) Length distribution of total sRNAs in LaSota-infected at different time points and mock infected HD11 cells; (B) analyses of the small RNA reads generated by
deep sequencing of 0 hpi, 24 hpi, and 48 hpi small RNA library. Figure 1. 2.1. Preliminary Analysis of Small RNA Sequencing Overview of the deep sequencing length data and cellular miRNA differential expression profile in chicken macrophages infected with the LaSota strain
Length distribution of total sRNAs in LaSota-infected at different time points and mock infected HD11 cells; (B) analyses of the small RNA reads generated by
sequencing of 0 hpi 24 hpi and 48 hpi small RNA library
Figure 1. Overview of the deep sequencing length data and cellular miRNA differential expression profile in chicken macrophages infected with the LaSota strain. (A) Length distribution of total sRNAs in LaSota-infected at different time points and mock infected HD11 cells; (B) analyses of the small RNA reads generated by
deep sequencing of 0 hpi, 24 hpi, and 48 hpi small RNA library. Figure 1. Overview of the deep sequencing length data and cellular miRNA differential expression profile in chicken macrophages infected with the LaSota strai
Length distribution of total sRNAs in LaSota-infected at different time points and mock infected HD11 cells; (B) analyses of the small RNA reads generated by
sequencing of 0 hpi 24 hpi and 48 hpi small RNA library
Figure 1. Overview of the deep sequencing length data and cellular miRNA differential expression profile in chicken macrophages infected with the LaSota strain. (A) Length distribution of total sRNAs in LaSota-infected at different time points and mock infected HD11 cells; (B) analyses of the small RNA reads generated by
deep sequencing of 0 hpi, 24 hpi, and 48 hpi small RNA library. 4 of 14 Pathogens 2019, 8, 123 2.2. Differentially Expressed miRNAs in Chicken Macrophages Infected with NDV Vaccine Strain LaSota 2.2. Differentially Expressed miRNAs in Chicken Macrophages Infected with NDV Vaccine Strain LaSota To identify miRNAs associated with NDV infection in chicken macrophages, miRNAs that were
differentially expressed were identified based on a p value < 0.05 and a |log 2 (fold change)| > 1
as the cut-offvalues. As compared with 0 hpi, 112 miRNAs (including 25 known and 87 novel)
were differentially expressed in cells of 24 hpi, 58 (including 15 known and 43 novel) of which
were upregulated while 54 (including 10 known and 44 novel) were downregulated; 115 miRNAs
(including 15 known and 100 novel) were differentially expressed in cells of 48 hpi, 76 (including
13 known and 63 novel) of which were upregulated while 39 (including 2 known and 37 novel) were
downregulated. 2.1. Preliminary Analysis of Small RNA Sequencing A total of 174 miRNAs were differentially expressed in cells of 48 hpi compared with
24 hpi, 97 (including 29 known and 68 novel) of which were upregulated while 77 (including 9 known
and 68 novel) were downregulated. Meanwhile, 12 upregulated and 6 downregulated differentially
expressed (DE) miRNAs were observed in common at 24 and 48 hpi compared with 0 hpi (Table 1). In addition, we found that at 24 hpi and 48 hpi, 46 and 64 DE miRNAs were uniquely upregulated, and
48 and 33 were uniquely downregulated compared with 0 hpi, respectively. Therefore, these findings
suggested that different miRNA expression characteristics exist among the NDV vaccine strain LaSota
at different post-infection times in chicken macrophages. To validate the sequencing data, six DE miRNAs (including three upregulated and three
downregulated miRNAs) were selected for qRT-PCR analysis. The results confirmed the upregulation
of expression of three miRNAs (gga-novel-800-mature, gga-novel-862-mature, and gga-miR-6606-5p)
and the downregulation of expression of three miRNAs (gga-novel-880-mature, gga-novel-20-mature,
and gga-novel-892-mature) in NDV-infected cells as compared with the mock infected cells. Thus, the
expression profiles obtained from the deep sequencing method were reliable and suitable for use in
further analysis. 5 of 14 Pathogens 2019, 8, 123 Table 1. Detailed information on 18 common differentially expressed (DE) miRNAs that were up-/down-regulated in groups of 24 h post inoculation (hpi) vs. 0 hpi
and 48 hpi vs. 0 hpi detected by deep sequencing methods. Number
Id
Sequences (5′–3′)
Regulated
24 hpi vs. 0 hpi
48 hpi vs. 2.1. Preliminary Analysis of Small RNA Sequencing As a result, a total of 7506
unique putative genes were targeted by 97 DE miRNAs between cells at 24 hpi and 0 hpi (Table S1),
8539 unique putative genes were targeted by 108 DE miRNAs between cells at 48 hpi and 0 hpi (Table
S2), and 8306 unique putative genes were targeted by 121 DE miRNAs between cells at 48 hpi and 24
hpi (Table S3). Our results, in agreement with those of other investigators [16,17], show that a single
miRNA molecule can regulate multiple genes. Conversely, multiple miRNAs can regulate a single
target gene, indicating that the miRNA-based regulatory network is extremely complicated. Next, these mRNA targets were sorted by the enrichment of GO (gene ontology) categories and
mainly clustered into different functional groups. The distribution of GO terms in the biological
process, cellular component, and molecular function among 0 hpi, 24 hpi, and 48 hpi as shown in
Figure 2. We found that, in all three libraries, the biological processes with the highest target numbers
were cellular processes (62%) (GO:0009987), single-organism processes (52%) (GO:0044699), metabolic
processes (45%) (GO:0008152), and biological regulation (45%) (GO:0065007); cell (61%) (GO:0005623),
organelle (45%) (GO:0043226), membrane (36%) (GO:0016020), and macromolecular complexes (20%)
(GO:0032991) were among the most abundant classes in the molecular function category; the common
cellular component terms were binding (50%) (GO:0005488), catalytic activity (26%) (GO:0003824),
molecular transducer activity (5%) (GO:0060089), and signal transducer activity (5%) (GO:0004871)
(Table S4). Meanwhile, these targets were functionally analyzed by KEGG (Kyoto Encyclopedia of Genes
Genomes) pathway annotation; 162, 161, and 162 KEGG pathways were obtained for the groups of
24 hpi vs. 0 hpi, 48 hpi vs. 0 hpi, and 48 hpi vs. 24 hpi. It was evident from our observations that a
large number of enriched genes were mainly involved in the mitogen-activated protein kinase (MAPK)
signaling pathway (gga04010), focal adhesion (gga04510), mTOR signaling pathway (gga04150),
and herpes simplex infection (gga05168), which were observed between cells at 24 hpi and 0 hpi, while
the most abundant KEGG terms corresponded to the MAPK signaling pathway, focal adhesion, protein
processing in endoplasmic reticulum (gga04141), and cellular senescence (gga04218), which were
present between cells at 48 hpi and 0 hpi. Moreover, the most abundant of the KEGG terms corresponded
to the MAPK signaling pathway, focal adhesion, tight junction (gga04530), cellular senescence, and
protein processing in endoplasmic reticulum were obtained between cells at 48 hpi and 24 hpi. 2.1. Preliminary Analysis of Small RNA Sequencing 0 hpi
Log2 FC
p Value
Log2 FC
p Value
1
gga-novel-1170-mature
UAUGGGAGGAACUGAAUGACAUG
up
2.05
1.24 × 10−2
1.82
1.63 × 10−2
2
gga-novel-800-mature
UGGGCGGCUGCGGGAGGG
up
3.76
1.27 × 10−3
3.10
2.11 × 10−2
3
gga-miR-6606-5p
GAGGAGCGGGAGGAGCGGGA
up
1.45
1.36 × 10−2
1.53
7.19 × 10−3
4
gga-novel-743-mature
UGGGGGUGCAGGUGGGGGGCU
up
1.68
4.81 × 10−2
2.34
4.74 × 10−3
5
gga-novel-138-mature
ACGGGACGGGGCGGGACGGCGC
up
1.36
3.31 × 10−2
1.20
4.87 × 10−2
6
gga-novel-862-mature
GAGCAAGGUACGGGGGGGU
up
2.32
3.27 × 10−4
2.09
1.00 × 10−3
7
gga-novel-904-mature
AGCAGAGAGAAGGGAUGAGGCU
up
1.57
1.16 × 10−2
1.24
4.15 × 10−2
8
gga-novel-417-mature
UGCUGGUAGGGGCCGACGACC
up
2.46
9.16 × 10−3
2.49
1.15 × 10−2
9
gga-novel-951-mature
AUGGAGGCGUGGGUUUUU
up
3.21
9.57 × 10−6
3.50
3.59 × 10−7
10
gga-novel-200-star
UGGGGAGGCCGCAGUGCAGGGCAA
up
2.99
1.02 × 10−2
2.63
2.22 × 10−2
11
gga-miR-1584
CCGGGUGGGGCUGGGCUGGG
up
2.36
9.62 × 10−3
3.22
1.10 × 10−4
12
gga-novel-672-mature
CCGCGGGGUGGGCGGGGGGCG
up
2.10
7.98 × 10−3
2.44
6.66 × 10−4
13
gga-novel-880-mature
UGCCGCUGCCCGGUGCUCACACU
down
−3.79
2.28 × 10−3
−2.84
1.14 × 10−2
14
gga-novel-226-mature
CGCAGCUCCGUUCCGUCCCCG
down
−1.62
2.75 × 10−2
−1.41
4.23 × 10−2
15
gga-novel-91-mature
UCCGCAGCUCCACUCCUGUCAC
down
−1.25
4.03 × 10−2
−2.23
3.67 × 10−4
16
gga-novel-20-mature
GCUCCUGCCUGGCUCGCCA
down
−3.64
1.71 × 10−8
−1.31
3.19 × 10−2
17
gga-novel-892-mature
UGCCGCUGCCCGGUGCUCACACU
down
−3.79
2.28 × 10−3
−2.84
1.14 × 10−2
18
gga-novel-95-mature
CUGCACUGCCACGCCGCGUUCC
down
−1.30
2.07 × 10−2
−1.13
4.48 × 10−2 Table 1. Detailed information on 18 common differentially expressed (DE) miRNAs that were up-/down-regulated in groups of 24 h post inoculation (hpi) vs. 0 hpi
and 48 hpi vs. 0 hpi detected by deep sequencing methods. Pathogens 2019, 8, 123 6 of 14 2.3. Target Gene Prediction and Functional Annotation Analyses for DE miRNAs after NDV Infection in
Chicken Macrophages 2.3. Target Gene Prediction and Functional Annotation Analyses for DE miRNAs after NDV Infection in
Chicken Macrophages .3. Target Gene Prediction and Functional Annotation Analyses for DE miRNAs after NDV Infection in
Chicken Macrophages 2.3. Target Gene Prediction and Functional Annotation Analyses for DE miRNAs after NDV Infection in
Chicken Macrophages To understand the molecular functions and biological processes of the DE miRNAs detailed in this
work, two independent algorithms, RNAhybrid and miRanda, were used to predict the mRNA targets
for each of the miRNAs that were significantly differentially expressed. 2.1. Preliminary Analysis of Small RNA Sequencing As we
observed, at least 15 KEGG pathways, including 2714, 3086, and 2999 genes were shown to be closely
related to immune responses in the groups of 24 hpi vs. 0 hpi, 48 hpi vs. 0 hpi, and 48 hpi vs. 24 hpi
(Figure 3). Among all the DE miRNAs, there are 18 DE miRNAs that remain up-/down-regulated when
infected with the LaSota vaccine strain even as the infection time progresses. A total of 7181
unique putative genes were targeted by 18 DE miRNAs between cells infected with NDV and mock
infected cells (Figure 4A and Table S5), at least 1485 of which were immune-relevant genes and were
involved in immune-related pathways (Figure 4B and Table S6), such as cellular senescence, the
cytokine–cytokine receptor interaction, the MAPK signaling pathway, RIG-I-like receptor signaling
pathway, apoptosis, necroptosis, the NOD-like receptor signaling pathway, Toll-like receptor signaling
pathway, and endocytosis. For example, the MAPK signaling pathway, which is involved in both
innate and adaptive immune responses [18,19], was represented by 120 genes and contained at least
four distinctly regulated groups of MAPKs, Jun amino-terminal kinases (JNK1/2/3), extracellular
signal-related kinases (ERK)-1/2, ERK5, and p38 proteins (p38 alpha/beta/gamma/delta) (Figure 5 and
Table S7). As was observed, the central proteins, MEKK1, MEKK4, MEK2, and c-Myc, within the MAPK
signaling pathway were regulated by gga-novel-743-mature, gga-novel-138-mature, gga-miR-1584,
gga-novel-800-mature, gga-novel-200-star, gga-novel-672-mature, and gga-novel-226-mature. Pathogens 2019, 8, 123 7 of 14 Pathogens 2019, 8, 123 Although mitochondrial antiviral-signaling protein (MAVS; also known as IPS-1, VISA, Cardif) is
a central adaptor protein in retinoic acid-inducible gene I (RIG-I)-mediated antiviral innate immunity
in mammals and birds (for example, goose MAVS is a vital antiviral that acts by activating type I
interferon when infected with NDV virulent strain Herts33/56 [20]), very little is known about the
mechanisms for miRNA regulation of type I interferon induction in NDV infection. In the present
study, we found that the essential adaptor protein MAVS was annotated as a target by downregulated
gga-miR-6609-3p, gga-novel-103-mature, and gga-novel-328-mature. Meanwhile, a previous study
suggested that NDV can activate the NOD-like receptor protein 3 (NLRP3) inflammasome [21], a critical
contributor in inflammatory responses; however, the detailed mechanism remains unknown. In this
study, NLRP3 was targeted by upregulated DE miRNA, such as gga-novel-200-star, gga-novel-36-star,
and gga-novel-424-mature. However, identification of DE cellular miRNA is just the first step towards
understanding miRNA regulation of interactions between the host and viral infection. 2.1. Preliminary Analysis of Small RNA Sequencing Then, more
works were planned to uncover the underlying mechanisms of how these potential genes are targeted
by DE miRNA and their roles in the live, low-virulent vaccine strain LaSota mediated immune cell
responses from the sides of host innate immune cells, such as chicken macrophages in the current study. Although mitochondrial antiviral-signaling protein (MAVS; also known as IPS-1, VISA, Cardif) is
a central adaptor protein in retinoic acid-inducible gene I (RIG-I)-mediated antiviral innate immunity
in mammals and birds (for example, goose MAVS is a vital antiviral that acts by activating type I
interferon when infected with NDV virulent strain Herts33/56 [20]), very little is known about the
mechanisms for miRNA regulation of type I interferon induction in NDV infection. In the present
study, we found that the essential adaptor protein MAVS was annotated as a target by downregulated
gga-miR-6609-3p, gga-novel-103-mature, and gga-novel-328-mature. Meanwhile, a previous study
suggested that NDV can activate the NOD-like receptor protein 3 (NLRP3) inflammasome [21], a critical
contributor in inflammatory responses; however, the detailed mechanism remains unknown. In this
study, NLRP3 was targeted by upregulated DE miRNA, such as gga-novel-200-star, gga-novel-36-star,
and gga-novel-424-mature. However, identification of DE cellular miRNA is just the first step towards
understanding miRNA regulation of interactions between the host and viral infection. Then, more
works were planned to uncover the underlying mechanisms of how these potential genes are targeted
by DE miRNA and their roles in the live, low-virulent vaccine strain LaSota mediated immune cell
responses from the sides of host innate immune cells, such as chicken macrophages in the current study. A previous study demonstrated that 61 DE miRNAs (including 36 upregulated and
25 downregulated miRNAs) were observed in visceral tissues from LaSota-infected specific-pathogen-free
(SPF) chicken embryos for 36 h as compared to the mock infected sample [22]; however, a much greater
number of DE miRNA (112 and 115 DE miRNA at 24 hpi and 48 hpi) were obtained in the chicken
macrophage, an innate immune cell, and target cell for NDV infection in vivo. Therefore, the finding
presented here also revealed that different miRNA regulation profiles exist among different cell lines
infected with the same virus. 2.1. Preliminary Analysis of Small RNA Sequencing A previous study demonstrated that 61 DE miRNAs (including 36 upregulated and
25 downregulated miRNAs) were observed in visceral tissues from LaSota-infected specific-pathogen-free
(SPF) chicken embryos for 36 h as compared to the mock infected sample [22]; however, a much greater
number of DE miRNA (112 and 115 DE miRNA at 24 hpi and 48 hpi) were obtained in the chicken
macrophage, an innate immune cell, and target cell for NDV infection in vivo. Therefore, the finding
presented here also revealed that different miRNA regulation profiles exist among different cell lines
infected with the same virus. 8 of 14
of 15 Pathogens 2019, 8, 123
Pathogens 2019, 8, x FO Figure 2. Gene ontology (GO) classification of miRNA-targeted mRNAs into three categories:
Molecular function, cell component, and biological process between cells at 24 hpi and 0 hpi
(A); between the cells at 48 hpi and 0 hpi (B); and between the cells at 48 hpi and 24 hpi (C). Figure 2. Gene ontology (GO) classification of miRNA-targeted mRNAs into three categories: Molecular
function, cell component, and biological process between cells at 24 hpi and 0 hpi (A); between the cells
at 48 hpi and 0 hpi (B); and between the cells at 48 hpi and 24 hpi (C). Figure 2. Gene ontology (GO) classification of miRNA-targeted mRNAs into three categories:
Molecular function, cell component, and biological process between cells at 24 hpi and 0 hpi
Figure 2. Gene ontology (GO) classification of miRNA-targeted mRNAs into three categories: Molecular
function, cell component, and biological process between cells at 24 hpi and 0 hpi (A); between the cells
at 48 hpi and 0 hpi (B); and between the cells at 48 hpi and 24 hpi (C). 9 of 14 Pathogens 2019, 8, 123 Figure 3 Number of genes in 12 immune relevant pathways annotated after infection with the LaSota vaccine strain
Figure 3. Number of genes in 12 immune-relevant pathways annotated after infection with the LaSota vaccine strain. Figure 3. Number of genes in 12 immune-relevant pathways annotated after infection with the LaSota vaccine strain. Figure 3. Number of genes in 12 immune-relevant pathways annotated after infection with the LaSota vaccine strain. Figure 3. Number of genes in 12 immune-relevant pathways annotated after infection with the LaSota vaccine strain. Figure 3. 2.1. Preliminary Analysis of Small RNA Sequencing Number of genes in 12 immune-relevant pathways annotated after infection with the LaSota v mber of genes in 12 immune-relevant pathways annotated after infection with the LaSota vaccine strain. 10 of 14 Pathogens 2019, 8, 123
Pathogens 2019, 8, x FOR Figure 4. DE miRNAs targets regulatory genes in pathways between samples infected with LaSota at different time points. (A) 18 common DE miRNA-
targets regulatory networks including 162 and 161 Kyoto Encyclopedia of Genes Genomes (KEGG) pathways in groups of 24 hpi vs 0 hpi and 48 hpi
vs 0 hpi; (B) nine immune-related pathways were selected to explore the targeting relationship between 15 of 18 DE miRNAs and immune factors. The red triangle represents up-regulated miRNAs and the green circle represents down-regulated miRNAs. Figure 4. DE miRNAs targets regulatory genes in pathways between samples infected with LaSota at different time points. (A) 18 common DE miRNA-targets
regulatory networks including 162 and 161 Kyoto Encyclopedia of Genes Genomes (KEGG) pathways in groups of 24 hpi vs 0 hpi and 48 hpi vs 0 hpi; (B) nine
immune-related pathways were selected to explore the targeting relationship between 15 of 18 DE miRNAs and immune factors. The red triangle represents
up-regulated miRNAs and the green circle represents down-regulated miRNAs. Figure 4. DE miRNAs targets regulatory genes in pathways between samples infected with LaSota at different time points. (A) 18 common DE miRNA-
targets regulatory networks including 162 and 161 Kyoto Encyclopedia of Genes Genomes (KEGG) pathways in groups of 24 hpi vs 0 hpi and 48 hpi
vs 0 hpi; (B) nine immune-related pathways were selected to explore the targeting relationship between 15 of 18 DE miRNAs and immune factors. The red triangle represents up-regulated miRNAs and the green circle represents down-regulated miRNAs. Figure 4. DE miRNAs targets regulatory genes in pathways between samples infected with LaSota at different time points. (A) 18 common DE miRNA-targets
regulatory networks including 162 and 161 Kyoto Encyclopedia of Genes Genomes (KEGG) pathways in groups of 24 hpi vs 0 hpi and 48 hpi vs 0 hpi; (B) nine
immune-related pathways were selected to explore the targeting relationship between 15 of 18 DE miRNAs and immune factors. The red triangle represents
up-regulated miRNAs and the green circle represents down-regulated miRNAs. 11 of 14 Pathogens 2019, 8, 123
Pathogens 2019, 8, x FOR Figure 5. The mitogen-activated protein kinase (MAPK) signaling pathway and related miRNAs. 3.1. In Vitro Infection of HD11 Cells with LaSota NDV vaccine strain LaSota (lentogenic, genotype II within class II, GenBank: AF077761) was
purchased from ATCC. The virus was propagated in 9–11-day-old SPF chicken embryos (MERIAL,
Beijing, China), and was purified directly from the infected allantois fluid, as described in previous
studies [10,23]. Chicken-origin macrophage HD11, a permanent chicken bone marrow macrophages
cell line was used in numerous chicken innate immune cell models for avian viruses, including the
NDV vaccine strain LaSota [13], and was kindly gifted by Prof. Daxin Peng, Yangzhou University. The HD11 cells were cultured in DMEM/F-12 (Dulbecco’s Modified Eagle Medium/Nutrient Mixture
F-12) supplemented with 10% fetal bovine serum (FBS) (Gibco, Shanghai, China) and 1% antibiotic
(100 µg/mL streptomycin and 100 U/mL penicillin) (Gibco, Shanghai, China) at 37 ◦C with 5% CO2. When the number of cells grew by 80%, the cell plates were washed twice with phosphate-buffered
saline (PBS). Afterward, the cells were incubated for 2 h with LaSota at 2 multiplicity of infection
(MOI) and control groups were set up with DMEM serum-free medium. There were 3 set replicates
per group, respectively. After 2 h, the cells were washed twice with PBS, and then 1% antibiotic and
2% FBS DMEM medium was added to each well and cultured at 37 ◦C Total RNA was extracted at
24 hpi and 48 hpi, respectively. Each group was processed in triplicate and pooled separately for
subsequent total RNA extraction. Then, three independent repeats of each time point were performed
for deep sequencing. 2.1. Preliminary Analysis of Small RNA Sequencing The red triangle represents up-regulated miRNAs and the green
circle represents down-regulated miRNAs. Figure 5. The mitogen-activated protein kinase (MAPK) signaling pathway and related miRNAs. The red triangle represents up-regulated miRNAs and the green
circle represents down-regulated miRNAs. Figure 5. The mitogen-activated protein kinase (MAPK) signaling pathway and related miRNAs. The red triangle represents up-regulated miRNAs and the green
circle represents down-regulated miRNAs. Figure 5. The mitogen-activated protein kinase (MAPK) signaling pathway and related miRNAs. The red triangle represents up-regulated miRNAs and the green
circle represents down-regulated miRNAs. 12 of 14 Pathogens 2019, 8, 123 Pathogens 2019, 8, 123 3.3. Small RNA Library Preparation and Illumina Sequencing An amount of 2 µg total RNA per sample was used as input for small RNA library preparation. The total RNA from each sample was sequentially ligated to 3′ and 5′ small RNA adapters using T4
RNA ligase according the manufacturer’s instructions (Illumina® TruSeq® Small RNA Library Prep
Kit, Illumina). Next, cDNAs were synthesized through reverse transcription using M-MLV reverse
transcriptase and the cDNAs were amplified by PCR. Then, 18–30 nt fragments of total small RNA from
PCR products were isolated using PAGE. Subsequently, three small RNA libraries, which included two
NDV-infected samples and one mock infected sample, were sequenced on an Illumina HiSeq 2500
platform (Illumina, San Diego, CA, USA). 3.4. Quantitative Real-Time RT-PCR (qRT-PCR) To validate the small RNA sequencing data, qRT-PCR was performed to validate the expression
profile of selected DE miRNAs using Bulge-loop™miRNA qRT-PCR Primer Sets (one RT primer and a
pair of qPCR primers for each set) and performed on an ABI StepOne System (Applied Biosystems,
Warrington, UK). The gene U6 was used as the reference gene. The relative expression level of each
miRNA in the infected samples was calculated using the 2−∆∆Ct method [24]. 3.2. RNA Extraction Total RNA was extracted using Tripure (Roche, Mannheim, Germany) following the manufacturer’s
recommendations. The quantity and purity of RNA were determined by ND-1000 Nanodrop®
Spectrophotometer (Thermo Scientific, Waltham, MA, USA), at A260/A280 nm ≥2, and the quality and
concentration of RNA were determined by an Agilent 2100 Bioanalyzer. Only samples with an RNA
integrity number greater than 8 were used for sequencing. 3.3. Small RNA Library Preparation and Illumina Sequencing 3.5. Sequencing Data Analysis Sequences that match the miRBase database are known as mature miRNA and sequences that do
not match the database are novel miRNA sequences. miRNAs with a p value ≤0.05 and log2 Fold
Change (Log2 FC) ≥1 are defined as DE miRNAs. Meanwhile, the miRNA target prediction software, 13 of 14 13 of 14 Pathogens 2019, 8, 123 RNAhybrid and miRanda, were used to predict the potential targets of DE miRNAs. To determine
the primary function of predicted target genes regulated by DE miRNAs using GO and KEGG
functional classifications. Furthermore, the network regulated by DE miRNAs was visualized using the
Cytoscape software. The red triangle represents up-regulated miRNAs and the green circle represents
down-regulated miRNAs. 4. Conclusions In conclusion, we identified miRNA and demonstrated the characterization of cellular miRNA in
chicken macrophages infected with the NDV vaccine strain LaSota using deep sequencing methods. Sixteen novel DE miRNA (including ten upregulated and six downregulated DE miRNA, respectively)
and two known upregulated miRNA candidates were observed in common at 24 hpi and 48 hpi
compared with 0 hpi. In addition, the number of potential target genes related to immune-relevant
genes and pathways were obtained by GO enrichment and KEGG pathway analysis. Taken together,
while these results identify the DE miRNA profiles in chicken macrophages infected with the NDV
vaccine strain LaSota, further studies are needed to understand how the live, low-virulent vaccine
strain LaSota mediated innate immune responses from the side of chicken macrophages. Supplementary Materials: The following are available online at http://www.mdpi.com/2076-0817/8/3/123/s1. Table S1 List for targets of all DE miRNAs of 24 hpi vs. 0 hpi; Table S2 List for targets of all DE miRNAs of 48 hpi
vs. 0 hpi; Table S3 List for targets of all DE miRNAs of 48 hpi vs. 24 hpi; Table S4 GO classification for targets of all
DE miRNAs; Table S5 Detailed information on 18 DE miRNAs and their corresponding target genes; Table S6
Detailed information on 18 DE miRNAs and their corresponding immune-related target genes and pathways;
Table S7 Detailed information on MAPK signaling pathway and related DE miRNAs. Author Contributions: R.Y., X.L., J.M., X.Y., and Z.D. designed and performed the study, drafted the manusc
nd analyzed the data. J.M., R.Y., X.L., J.L., Y.F., Z.D., and X.Y. carried out experiments. Funding: This research was partly funded by two grants from the Key Project of Chinese National Programs
for Research and Development (2017YFD0500800 and 2018YFD051404), and one grant from the Natural Science
Foundation of Jilin Province (20190301087NY). Acknowledgments: This study was partly supported by two grants from the Key Project of Chinese National
Programs for Research and Development (2017YFD0500800 and 2018YFD051404), and one grant from the Natural
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НОВАЯ КОНСТИТУЦИЯ - СТАНОВИТСЯ ВЫСШИМ ПРИМЕРОМ СПРАВЕДЛИВОГО ПРАВОВОГО ТВОРЧЕСТВА В НОВОМ УЗБЕКИСТАНЕ
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М.Р.Анорбоев
|
Russian
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Spoken
| 955
| 3,006
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НОВА
ПР
Преподав
Академии
до
ARTICLE INFO
Received: 04th May 2023
Accepted: 12th May 2023
Online: 13th May 2023
KEY WORDS
Новая конституция, закон,
субъект
законодательной
инициативы,
нормотворчество,
правотворчество,
концепция. В
статье
рассматривается
принятие
Новой
Конституции,
законодательный
процесс,
высказанные учеными мнения относительно роли
Новой Конституции в жизни общества, изменения,
которые должны быть реализованы в результате
принятия Новой Конституции, обсуждены реформы,
реализуемые в законотворческом процессе, и их
последствия, его значение на сегодняшний день и
проблемы
адаптации
к
предусмотренным
Конституцией изменениям и их реализации. Верховенство закона служит установлению справедливости в любом обществе и
повышению
эффективности
правовых
реформ. Важнейшим
условием
демократического общества является то, чтобы законы, принимаемые в стране, были
справедливыми и отражали интересы народа. [1] Только при неукоснительном
соблюдении таких законов в обществе установится и укрепится демократия. Одним из
важнейших признаков демократического общества является равенство членов
общества перед законом, верховенство Конституции и законов. Конечной целью
конституции и законов является обеспечение прав и свобод человека. Поэтому
обеспечение законности, усиление защиты прав и интересов личности, семьи,
общества и государства, повышение правовой культуры и правосознания населения,
воспитание граждан в духе послушания и уважения к закону является подлинно
демократическим, правовым государством, основанным на развитой рыночной
экономике, и является не только целью построения свободного гражданского
общества, но и его средством, одним из важнейших условий. Конституция
является
основным
законом
любой
страны. Содержание
конституции состоит в том, чтобы определить все основы государственной,
общественной и политической жизни страны. Иными словами, он является основой и
фундаментом правовой системы, и на его основе строятся все остальные законы. Однако в то же время Конституция не является фиксированным, непреложным
документом. Если есть необходимость изменить Конституцию, значит, общество
вступает в новый этап развития, указывает на жизненную необходимость. В нашей обновленной Конституции мы видим, что каждая статья четко и
лаконично объяснена и проработана. В результате налаживания прямого диалога EURASIAN JOURNAL OF LAW, FINANCE AND
APPLIED SCIENCES
Innovative Academy Research Support Center
UIF = 8.3 | SJIF = 5.961
www.in-academy.uz www.in-academy.uz между народом и органами государственной власти в корне изменилось отношение к
изменениям, осуществляемым по инициативе органов государственной власти и
управления. Политика
прозрачности
привела
к
оперативному
реагированию
государственных органов на общественное мнение и оперативному решению
существующих социально-экономических проблем. Впервые в истории современного Узбекистана в конституционных реформах
активно участвует сам народ. Особое значение имеет и то, что с момента объявленной
конституционной реформы поступило более 300 000 предложений, [2] но самое
главное, что проект Конституционного Закона формировалось на основе этих
предложений, можно сказать, что все это означает, что Новая Конституция стала
буквально Народной Конституцией. Конституция Республики Узбекистан является высоким образцом политического
и правового мышления нашего народа. Это была правовая гарантия свободной и
независимой, мирной и мирной жизни, ни от кого не зависящей. Он служит прочной
основой для построения правового демократического государства, основанного на
рыночных отношениях и сильном гражданском обществе. Безусловно, не напрасно мы
связываем все достижения, которых мы достигли на нелегком и почетном пути, в
первую очередь, с нашей Конституцией. Ведь наш Основной закон определил те
гарантии нашей национальной независимости и развития, прав и свобод человека, о
которых наш народ мечтал долгие годы. На основе нашей конституции в нашей стране были сформированы национальная
правовая система, государственные органы, институты гражданского общества. Сегодня масштабные реформы осуществляются по всем направлениям. Растет наш
социально-экономический, политический и военный потенциал, все больше растет
мировоззрение наших граждан. Все это, прежде всего, результат живительной силы
нашего Генерального Совета. За прошедшую четверть века все коренные реформы и преобразования в нашей
стране осуществлялись на основе нашей Конституции. Это свидетельствует о том, что
это важный политический документ, полностью отвечающий интересам нашего
народа и стратегическим целям нашей страны. Здесь следует отметить, что большинство изменений в этом направлении следует
воспринимать как должное на основе реформ, проводимых нашим Президентом для
полной реализации приоритетного принципа «За достоинство человека».[3] Причина в
том, что Президент выдвинул идею о том, что принцип «человек-общество-
государство» должен быть положен в основу всех изменений нашего Основного закона
и должен быть отражен в содержании каждой нормы как основное направление
реализации конституционные реформы. Согласно нашей недавно принятой Конституции, предложено: «Никто не может
быть лишен своего имущества без решения суда. [4] главное, что на этой основе в нашей стране создана уникальная национальная
система обеспечения и защиты прав человека и созданы специальные в этом вопросе
национальные
институты. Узбекистан
получил
на
основании
декларации
ратификацию международных конвенций, принятых в области права человека, намоен EURASIAN JOURNAL OF LAW, FINANCE AND
APPLIED SCIENCES
Innovative Academy Research Support Center Innovative Academy Research Support Center
UIF = 8.3 | SJIF = 5.961
www.in-academy www.in-academy.uz продолжает свою приверженность общепринятым принципам. Эти усилия, особенно
нашего президента Шавката Мирзиеева, были предприняты ранее, наш народ занял
прочное место в их сердцах и стали нашим ежедневным девизом "Люди - это не
государственные учреждения, государственные учреждения служат нашему народу
надо делать!", [5] "Прежде всего, чтобы наши люди ощутили эффект реформ,
необходимо, чтобы они чувствовали себя в своей тарелке не в будущем, а сегодня!",
"Новые Давайте жить свободно и комфортно в Узбекистане!", [6] в дальнейшем
развивающийся на основе благородных идей. Вместо заключения, будущее демократического общества, к которому мы
стремимся, прежде всего, демократические ценности находятся в сердцах каждого из
нас, и насколько глубоко они занимают место в нашем сознании, напрямую зависит от
того, насколько хорошо эти ценности сохраняются, развиваются и передаются
будущим поколениям. В связи с этим каждый из нас задумывается о сути, значении нашей новой
Конституции глубокое понимание и строгое следование новым основополагающим
требованиям гуманности - наш долг. References: 1. Саидов А.Х. Ўзбекистон Конституцияси тарихи.-Т.:2018 –Б.346 2. https://saylov.uz/oz 3. Выступление вновь избранного Президента Узбекистана Шавката Мирзиѐева на
церемонии инаугурации -https://review.uz/post/vstuplenie-vnov-izbrannogo-prezidenta-
uzbekistana-shavkata-mirziyoyeva-na-ceremonii-inauguracii 3. Выступление вновь избранного Президента Узбекистана Шавката Мирзиѐева на
церемонии инаугурации -https://review.uz/post/vstuplenie-vnov-izbrannogo-prezidenta-
uzbekistana-shavkata-mirziyoyeva-na-ceremonii-inauguracii 4. https://daryo.uz/2022/08/04/shaxs-oz p //
y
/
/
/
/
5. https://www.norma.uz/oz/bizning_sharhlar/prezident_konstituciyaga_bir_qator_uzgarti
rishlar_kiritishni_taklif_qild
6. https://president.uz/ru/lists/view/1328 p //
y
/
/
/
/
5. https://www.norma.uz/oz/bizning_sharhlar/prezident_konstituciyaga_bir_qator_uzgarti
rishlar_kiritishni_taklif_qild
6
htt
//
id
t
/
/li t / i
/1328 q
6. https://president.uz/ru/lists/view/1328 6. https://president.uz/ru/lists/view/1328 Volume 3 Issue 5, May 2023 Page 138
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https://openalex.org/W3112790134
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OpenAlex
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Open Science
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CC-By
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Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort: Comparison of a food frequency questionnaire with plasma phospholipid marine n-3 levels
|
Joe Chan
|
English
|
Spoken
| 7,581
| 14,190
|
Background High levels of plasma marine n-3 fatty acids (n-3FAs) are associated with improved patient
and graft survival in renal transplant recipients (RTRs). The aim of this study was to evaluate
the utility of a new food frequency questionnaire (FFQ) to estimate marine n-3FA consump-
tion in future epidemiological research. Editor: Stefano Turolo, Ospedale Maggiore
Policlinico, ITALY Editor: Stefano Turolo, Ospedale Maggiore
Policlinico, ITALY
Received: April 29, 2020
Accepted: December 2, 2020
Published: December 17, 2020 Methods Received: April 29, 2020
Accepted: December 2, 2020
Published: December 17, 2020 We developed an FFQ with a simple design of 10 questions to assess intake of marine
sources of n-3FAs. RTRs included in the recent ORENTRA trial (n = 132) completed the
study FFQ at the baseline visit eight weeks after engraftment and at the end of study visit
one year post-transplant. We measured the reference biomarker plasma phospholipid (PL)
marine n-3FA levels by gas chromatography at the same time points to evaluate association
and degree of agreement between FFQ based marine n-3FA consumption estimates and
the biomarker. Peer Review History: PLOS recognizes the
benefits of transparency in the peer review
process; therefore, we enable the publication of
all of the content of peer review and author
responses alongside final, published articles. The
editorial history of this article is available here:
https://doi.org/10.1371/journal.pone.0244089 Marine n-3 fatty acid consumption in a
Norwegian renal transplant cohort:
Comparison of a food frequency
questionnaire with plasma phospholipid
marine n-3 levels Joe ChanID1,2*, My Svensson1,2, Trond Jenssen2,3, Erik B. Schmidt4, Ivar A. Eide1,3 a1111111111
a1111111111
a1111111111
a1111111111
a1111111111 a1111111111
a1111111111
a1111111111
a1111111111
a1111111111 1 Department of Renal Medicine, Akershus University Hospital, Lørenskog, Norway, 2 Institute of Clinical
Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway, 3 Department of Transplantation Medicine,
Oslo University Hospital, Rikshospitalet, Oslo, Norway, 4 Department of Clinical Medicine, Aalborg University
Hospital, Aalborg, Denmark * joe.chan@ahus.no OPEN ACCESS Citation: Chan J, Svensson M, Jenssen T, Schmidt
EB, Eide IA (2020) Marine n-3 fatty acid
consumption in a Norwegian renal transplant
cohort: Comparison of a food frequency
questionnaire with plasma phospholipid marine n-3
levels. PLoS ONE 15(12): e0244089. https://doi. org/10.1371/journal.pone.0244089 PLOS ONE PLOS ONE PLOS ONE PLOS ONE Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort Funding: The author(s) received no specific
funding for this work. Conclusions Funding: The author(s) received no specific
funding for this work. Marine n-3FA consumption estimates based on the FFQ showed a moderate correlation
with the reference biomarker plasma PL marine n-3FA levels. The FFQ might be useful in
epidemiological studies where resources are limited. Competing interests: The authors have declared
that no competing interests exist. Introduction Marine n-3 fatty acid (n-3FA) consumption may benefit cardiovascular health and renal func-
tion following renal transplantation [1, 2]. Previous clinical trials in renal transplant recipients
(RTRs) report lower triglyceride levels, higher high-density lipoprotein cholesterol levels and
lower diastolic blood pressure after marine n-3FA supplementation [1]. A large cohort study
in Norwegian RTRs showed that high plasma phospholipid (PL) n-3FA levels were associated
with improved patient and graft survival [3, 4]. Antifibrotic and renoprotective effects of long-
term high-dose marine n-3FA supplementation have also been shown for other cardiovascular
high-risk populations like myocardial infarction survivors [5, 6]. The recent “Omega-3 fatty
acids in Renal Transplantation (ORENTRA)” trial performed in Norwegian RTRs found lower
levels of inflammatory biomarkers, less development of renal graft fibrosis and improvement
of endothelial function, as well as reduced triglyceride levels after 44 weeks of high-dose n-3FA
supplementation [2]. Observational studies and randomized clinical trials (RCTs) studying the influence of
marine n-3FA intake on cardiovascular health report conflicting results [7–14]. But a recent
meta-analysis, which included three recent large RCTs [15–17], concluded that marine n-3FA
supplementation was associated with a lower risk of cardiovascular events and death [18]. In
renal transplantation, further studies are warranted to evaluate to what extent marine n-3FA
consumption may improve patient and graft survival. The major marine n-3FAs eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)
are found in fish and other seafood. Plasma PL levels of EPA and DHA can be measured by
fatty acid analysis and are considered valid and reliable measures of marine n-3FA consump-
tion [19]. However, fatty acid analysis is more expensive and time-consuming to apply than a
food frequency questionnaire (FFQ). Hence, replacing fatty acid analyses with an FFQ focused
on marine n-3FA consumption seems attractive in epidemiological research, provided that the
FFQ values show a high degree of agreement and association with the reference biomarker. g
g
g
The main objective of this study was therefore to evaluate the utility of a new FFQ focused
on marine n-3FA consumption, using plasma PL marine n-3FA level as the reference
biomarker. Results The median plasma PL marine n-3FA level was 6.0 weight percentage (wt)% (interquartile
range [IQR] 4.7 to 7.3) at baseline and 6.3 wt% (IQR 4.8 to 7.4) at end of study. Median FFQ
based marine n-3FA consumption estimates were 22.8 g/month (IQR 13.0 to 34.0) at base-
line and 20.3 g/month (IQR 14.5 to 32.3) at end of study. FFQ based marine n-3FA con-
sumption estimates showed a moderate correlation with plasma PL marine n-3FA levels at
baseline (Spearman’s correlation coefficient rs = 0.43, p<0.001) and a stronger correlation
at end of study (rs = 0.62, p<0.001). Bland Altman plots showed a reasonable degree of
agreement between the two methods at both time points. Copyright: © 2020 Chan et al. This is an open
access article distributed under the terms of the
Creative Commons Attribution License, which
permits unrestricted use, distribution, and
reproduction in any medium, provided the original
author and source are credited. Data Availability Statement: All relevant data are
within the paper and its Supporting Information
files. 1 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 Study participants and design The study cohort consisted of 132 adult Norwegian RTRs included in the ORENTRA trial [2],
who were randomized to receive daily supplementation of either 2.6 g of marine n-3FAs (EPA
plus DHA) or 3 g of extra virgin olive oil (control oil) for 44 weeks. All patients gave written
informed consent for participation in the trial, which also comprised the study FFQ and fatty
acid analysis. The study was approved by the Regional Committees for Medical and Health
Research Ethics in Norway and was performed in accordance with the Declaration of Helsinki
(Clinical.Trials.gov identifier NCT01744067). FFQ and fatty acid analysis were performed 2 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 PLOS ONE Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort eight weeks post-engraftment (baseline visit) and one year after transplantation (end of study
visit). Patients were treated with standard triple maintenance immunosuppressive regimen
consisting of prednisolone, mycophenolate and tacrolimus. Blood samples were drawn in a
fasting state in the morning at the baseline and end of study visits. Gas chromatography was
used to determine individual fatty acid levels in plasma PLs, quantified as weight percentage
(wt%) of total plasma PL fatty acids. We defined marine n-3FA level as the sum of EPA and
DHA. The study was performed at Oslo University Hospital during 2012–2015. Details regard-
ing recruitment of patients, fatty acid analysis and the study FFQ are provided in the S1 File. For the ORENTRA trial, we developed a specific FFQ with a simple design of 10 multiple-
choice questions (Fig 1), focusing on food items containing marine sources of n-3FA that are
typically found in a Nordic diet [20]. We used three different approaches to estimate marine n-3FA consumption at baseline and
end of study based on the FFQ recordings: 1. Marine n-3FA consumption estimates, calculated by combining data from the FFQ with
known content of EPA and DHA in fish and other seafoods [21], assuming a standard por-
tion size for a Norwegian population (S1 File, S2 Fig). 2. Marine n-3FA consumption estimates calculated as in approach 1 using only data on fatty
fish intake for lunch and dinner (S3 Fig). 3. Number of fish servings per month (S4 Fig). Statistical analysis We used correlation analysis (Spearman’s correlation coefficient [rs]) and multivariate regres-
sion analysis (data presented as standardized regression coefficients [Std. β-coeff.]) to study
associations between FFQ based marine n-3FA consumption estimates and plasma PL marine
n-3FA levels. Data obtained by the reference biomarker and the study FFQ were standardized
using z-statistics to produce data for both methods on the same scale. This allowed for a more
meaningful visual presentation (scatter plots) and made it possible to analyze degree of agree-
ment using Bland Altman plots and one-sample t-test. Since the study drug used in the OREN-
TRA trial was high-dose marine n-3FA supplementation, we excluded patients in the
intervention group when performing statistical analysis of data from the end of study visit. Two patients belonging to the control group did not meet at the end of study visit (n = 66 at
baseline, n = 64 at end of study visit). Patient characteristics at baseline grouped according to
plasma PL marine n-3FA tertiles were evaluated with analysis of variance for continuous data
and Mantel-Haenszel linear-by-linear-trend for categorical data. A two-sided p-value
of < 0.05 was considered statistically significant. We used SPSS1 version 25.0 (IBM, New
York, NY, US) for statistical analyses. https://doi.org/10.1371/journal.pone.0244089.g001 Results Patient characteristics for the study cohort have previously been published in detail [2]. Selected variables, grouped according to plasma PL n-3FA tertiles at baseline eight weeks
post-transplant, are presented in Table 1. Patients in the upper tertile were older and less often
current smokers. Supplementation with cod liver oil was used by 28% of patients in the upper
tertile compared with 9% in the lower. Median plasma PL n-3FA levels were 6.0 wt% (inter-
quartile range [IQR] 4.7 to 7.3, n = 132) at the baseline visit and 6.3 wt% (IQR 4.8 to 7.4,
n = 64) at the end of study visit. Median FFQ based marine n-3FA consumption estimates
were 22.8 g/month (IQR 13.0 to 34.0, n = 132) at baseline and 20.3 g/month (IQR 14.5 to 32.3, PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 3 / 14 PLOS ONE Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort Fig 1. Study food frequency questionnaire focusing on food items containing marine n-3 polyunsaturated fatty acids (English
version). The study subjects responded to the question “During a typical month, how often do you eat these food items?” using one of
six response alternatives for each of the ten food item categories. https://doi.org/10.1371/journal.pone.0244089.g001 PLOS ONE Table 1. Patient characteristics at baseline eight weeks after renal transplantation according to plasma phospholipid marine n-3 fatty acid tertiles. Variables
All patients
Plasma PL marine n-3FA level, wt%
p (trend)
5.1
5.2–6.9
7.0
Number of patients
132
44
44
44
FFQ based marine fatty acid consumption estimate, g/month
26.0 (16.6)
19.8 (15.0)
25.4 (17.5)
33.1 (14.7)
0.001
Number of servings of fish / month
19.9 (15.6)
13.3 (13.8)
16.2 (13.4)
26.9 (16.8)
0.12
Marine n-3FA supplements, %
14.5
9.1
6.8
27.9
0.01
Recipient age, years
53.4 (13.8)
45.7 (12.6)
55.6 (13.6)
59.1 (12.0)
<0.001
Recipient gender (Female), %
25.8
27.3
25.0
25.6
0.86
Ethnicity, White, %
92.4
93.2
86.0
97.7
0.39
Body mass index, kg/m2
26.0 (3.9)
25.2 (4.0)
25.9 (3.8)
26.8 (3.7)
0.16
Educational level, %
>3 years at University
29.8
27.3
27.3
34.9
1–3 years at University
7.6
6.8
6.8
9.3
Secondary school
35.1
40.9
36.4
27.9
Primary school
27.5
25.0
29.5
27.9
0.56
Physical exercise, %
High intensity twice per week
42.1
51.2
36.4
39.0
High intensity once per week
9.5
2.4
11.4
14.6
Low intensity twice per week
34.9
36.6
36.4
31.7
Low intensity once per week
7.1
9.8
9.1
2.4
None
6.3
0.0
6.8
12.2
0.29
Smoking habits, %
Daily smoker
12.7
19.5
11.4
7.3
Non-daily smoker
3.2
7.3
0.0
2.4
Former heavy smoker
7.1
7.3
4.5
9.8
Former light smoker
35.7
34.1
43.2
29.3
Life-long non-smoker
41.3
31.7
40.9
51.2
0.03
Patient characteristics are presented as percentage for categorical data and mean value (standard deviation) for continuous variables. Differences between groups were
analyzed by analysis of variance and Mantel Haenszel linear-by-linear trend as appropriate. Patient characteristics are presented as percentage for categorical data and mean value (standard deviation) for continuous variables. Differences between groups were
analyzed by analysis of variance and Mantel Haenszel linear-by-linear trend as appropriate. https://doi.org/10.1371/journal.pone.0244089.t001 n = 64) at end of study. Marine n-3FA consumption remained stable during follow-up in the
control group with a median increase of plasma PL marine n-3FA level of 0.1 wt% (IQR -0.8 to
1.0) and change in FFQ based marine n-3FA consumption estimates of -1.0 g/month (IQR
-9.4 to 6.3). At baseline, moderate correlations were found between FFQ based marine n-3FA con-
sumption estimates and the reference biomarker plasma PL marine n-3FA levels (approach 1,
rs = 0.43, p<0.001, n = 132, Fig 2). Fig 1. Study food frequency questionnaire focusing on food items containing marine n-3 polyunsaturated fatty acids (English
version). The study subjects responded to the question “During a typical month, how often do you eat these food items?” using one of
six response alternatives for each of the ten food item categories. 4 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 PLOS ONE A reasonable degree of agreement between the study FFQ
estimates and the reference biomarker was shown in a Bland Altman plot (Fig 3) and signifi-
cant bias was ruled out by a one-sample t-test (t = 0.04, p = 0.96). Two groups of outlier obser-
vations were identified. One group consisted of patients reporting high intake of marine n-
3FAs but had average plasma PL marine n-3 FA levels. Another group with high or very high
plasma PL marine n-3FA levels had average marine n-3FA consumption according to the
study FFQ. Correlations between FFQ based marine n-3FA consumption estimates and plasma PL
marine n-3FA levels were stronger at the end of study (rs = 0.60, p<0.001, n = 64, Fig 4) than
at baseline. One-sample t-test (t = 0.06, p = 0.95) and a Bland Altman plot confirmed an
acceptable degree of agreement between the two methods at this time point (Fig 5). 5 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 PLOS ONE Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort Fig 2. Scatterplots of standardized plasma PL marine n-3FA levels and standardized FFQ based marine n-3FA
consumption estimates with regression line at eight weeks post-transplant (n = 132). https://doi.org/10.1371/journal.pone.0244089.g002 Fig 2. Scatterplots of standardized plasma PL marine n-3FA levels and standardized FFQ based marine n-3FA
consumption estimates with regression line at eight weeks post-transplant (n = 132). Fig 2. Scatterplots of standardized plasma PL marine n-3FA levels and standardized FFQ based marine n-3FA
consumption estimates with regression line at eight weeks post-transplant (n = 132). https://doi.org/10.1371/journal.pone.0244089.g002 https://doi.org/10.1371/journal.pone.0244089.g002 Baseline correlation analysis was repeated for patients belonging to the ORENTRA trial
control group (rs = 0.45, p<0.001, n = 66, S5 Fig) and we found a high degree of agreement
between the methods (S6 Fig), similar to what was shown for the whole study cohort at
baseline. We performed a multivariate stepwise forward regression analysis, adjusting for the poten-
tial confounding factors recipient age, gender, height, weight, body mass index, renal function,
physical activity, educational level and smoking habits (p<0.10 for inclusion of variables in the
final regression model) at baseline and end of study. The reference biomarker plasma PL
marine n-3FA level was associated with FFQ based marine n-3FA consumption estimates (Std. β-coeff. 0.24, p = 0.01), as well as recipient age (Std. β-coeff. 0.25, p = 0.01) and smoking habits
(Std. β-coeff. PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 PLOS ONE 0.15, p = 0.06) at baseline (n = 132). Together the three variables included in final Fig 3. Bland-Altman plot assessing degree of agreement between standardized plasma PL marine n-3FA levels
and standardized FFQ based marine n-3FA consumption estimates at baseline eight weeks post-transplant. We
used standardization of data obtained by the study FFQ and reference biomarker, hence the mean value was set at 0. The upper and lower limits of agreement were set at 2 standard deviations from the mean. The Bland Altman plot
includes all patients enrolled in the ORENTRA trial (n = 132) at the baseline time-point. https://doi.org/10.1371/journal.pone.0244089.g003 Fig 3. Bland-Altman plot assessing degree of agreement between standardized plasma PL marine n-3FA levels
and standardized FFQ based marine n-3FA consumption estimates at baseline eight weeks post-transplant. We
used standardization of data obtained by the study FFQ and reference biomarker, hence the mean value was set at 0. The upper and lower limits of agreement were set at 2 standard deviations from the mean. The Bland Altman plot
includes all patients enrolled in the ORENTRA trial (n = 132) at the baseline time-point. Fig 3. Bland-Altman plot assessing degree of agreement between standardized plasma PL marine n-3FA levels
and standardized FFQ based marine n-3FA consumption estimates at baseline eight weeks post-transplant. We
used standardization of data obtained by the study FFQ and reference biomarker, hence the mean value was set at 0. The upper and lower limits of agreement were set at 2 standard deviations from the mean. The Bland Altman plot
includes all patients enrolled in the ORENTRA trial (n = 132) at the baseline time-point. https://doi.org/10.1371/journal.pone.0244089.g003 6 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 PLOS ONE Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort Fig 4. Scatterplots of standardized plasma PL marine n-3FA levels and standardized FFQ based marine n-3FA
consumption estimates with regression line at one year post-transplant for patients belonging to the control
group of the ORENTRA trial (n = 64). Fig 4. Scatterplots of standardized plasma PL marine n-3FA levels and standardized FFQ based marine n-3FA
consumption estimates with regression line at one year post-transplant for patients belonging to the control
group of the ORENTRA trial (n = 64). https://doi.org/10.1371/journal.pone.0244089.g004 https://doi.org/10.1371/journal.pone.0244089.g004 regression model explained 23% of the variance in plasma PL marine n-3FA levels. At the end
of study, only FFQ based marine n-3FA consumption estimates (Std. β-coeff. PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 PLOS ONE 0.54, p<0.001)
was included in the final regression model, and it explained 29% of the variance in the refer-
ence biomarker. Correlations between FFQ based marine n-3FA consumption estimates and the reference
biomarker were slightly weaker for fatty fish intake (approach 2, baseline rs = 0.35 and end of
study rs = 0.46) and number of fish servings per month (approach 3, baseline rs = 0.38 and end
of study rs = 0.43) than for total marine n-3FA consumption estimates (approach 1). Correla-
tions with the reference biomarker for individual food items included in the study FFQ are
shown in S7 Fig. The food item cod liver oil showed a low correlation with the reference Fig 5. Bland-Altman plot assessing degree of agreement between standardized plasma PL marine n-3FA levels
and standardized FFQ based marine n-3FA consumption estimates at one year post-transplant. We used
standardization of data obtained by the study FFQ and reference biomarker, hence the mean value was set at 0. The
upper and lower limits of agreement were set at 2 standard deviations from the mean. The Bland Altman plot includes
only patients belonging to the control group of the ORENTRA trial (n = 64) at the end of study time-point. https://doi.org/10.1371/journal.pone.0244089.g005 Fig 5. Bland-Altman plot assessing degree of agreement between standardized plasma PL marine n-3FA levels
and standardized FFQ based marine n-3FA consumption estimates at one year post-transplant. We used
standardization of data obtained by the study FFQ and reference biomarker, hence the mean value was set at 0. The
upper and lower limits of agreement were set at 2 standard deviations from the mean. The Bland Altman plot includes
only patients belonging to the control group of the ORENTRA trial (n = 64) at the end of study time-point. https://doi.org/10.1371/journal.pone.0244089.g005 PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 7 / 14 PLOS ONE Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort biomarker at baseline (rs = 0.21). Marine n-3FA supplementation, including cod liver oil, was
discontinued after enrollment in the ORENTRA trial and consequently intake of cod liver oil
did not influence results at one year post-transplant. Discussion The main finding of the present study was that marine n-3FA consumption estimates based on
a new FFQ focused on fish consumption showed a moderate correlation with the reference
biomarker plasma PL marine n-3FA levels at eight weeks post-transplant and a slightly stron-
ger correlation at one year post-transplant. The correlations found in the present study are in
the range of what is regarded as acceptable in FFQ validation studies [22]. Previous FFQs have mainly focused on fatty fish intake, assumed to reflect marine n-3FA
consumption better than total fish intake [23–35]. We hypothesized that a more meticulous
approach using weighted response scales based on EPA and DHA content in fatty and lean
fish, other seafoods and marine n-3FA supplements would provide a more precise estimation
of marine n-3FA consumption. In our cohort, approach 1, which estimated total marine n-
3FA consumption from all the data obtained by the study FFQ, showed a stronger correlation
with the reference biomarker than approach 2 (which only focused on fatty fish intake) and 3
(which used the number of fish servings), suggesting that our hypothesis was correct. However, the study FFQ only provided slightly stronger correlations than most recent
FFQs focused on fish consumption (Table 2) and the utility of the study FFQ will have to be
confirmed by other studies before it can be used in epidemiological research. Plasma PL marine n-3FA levels did not differ between baseline and end of study visits for
the majority of patients. This is consistent with previous reports from large Norwegian cohorts
and supports the notion that a single fatty acid measurement may be acceptable for epidemio-
logical studies [3, 19]. However, the association between FFQ based marine n-3FA consump-
tion estimates and the reference biomarker was stronger at end of study than at baseline. There could be several explanations to this finding. Study participants might have become
more aware of their eating habits due to participation in the ORENTRAL trial and reported
fish consumption more accurately when they completed the FFQ the second time. We found a There could be several explanations to this finding. Study participants might have become
more aware of their eating habits due to participation in the ORENTRAL trial and reported
fish consumption more accurately when they completed the FFQ the second time. We found a Table 2. https://doi.org/10.1371/journal.pone.0244089.t002 Discussion Summary of selected food frequency questionnaire validation studies published during the last six years, focusing on fish and/or marine fatty acid con-
sumption, using circulating phospholipids or erytrocytes as the reference biomarker. uestionnaire validation studies published during the last six years, focusing on fish and/or marine fatty acid con-
ytrocytes as the reference biomarker. Table 2. Summary of selected food frequency questionnaire validation studies published during the last six years, focusing on fish and/or marine fatty acid con-
sumption, using circulating phospholipids or erytrocytes as the reference biomarker. First author (reference)
Published, year
n
Study population
Reference marine fatty acid biomarker
Correlation coefficient
Giovannelli J [23]
2014
2630
General population
Plasma phospholipid
r = 0.39–0.43
Lassale C [24]
2016
198
General population
Plasma phospholipid
rs = 0.51–0.54
Sluik D [25]
2016
383
General population
Plasma phospholipid
r = 0.43–0.47
Whitton C [26]
2017
161
General population
Plasma phospholipid
r = 0.36
Laursen UB [27]
2018
200
General population
Plasma phospholipid
rs = 0.45
Shen W [28]
2019
108
General population
Whole blood phospholipid
r = 0.67
Schumacher TL [29]
2016
39
Hyperlipidemia
Erythrocyte
rs = 0.53–0.62
Allaire J [30]
2015
60
Prostate cancer
Erythrocyte
rs = 0.59
Brunvoll SH [31]
2018
49
Breast cancer
Serum phospholipid
r = 0.36–0.53
Lepsch J [32]
2014
248
Pregnant women
Serum phospholipid
rs = 0.21–0.26
Zhou YB [33]
2017
804
Pregnant women
Plasma phospholipid
rs = 0.35
Erythrocyte
rs = 0.33
Kobayashi M [34]
2017
188
Pregnant women
Serum phospholipid
rs = 0.33–0.45
Liu MJ [35]
2016
408
Lactating women
Plasma phospholipid
rs = 0.36
Erythrocyte
rs = 0.24 Summary of selected food frequency questionnaire validation studies published during the last six years, focusing on fis
on, using circulating phospholipids or erytrocytes as the reference biomarker. PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 8 / 14 PLOS ONE Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort lower correlation with the reference biomarker for cod liver oil than for other food items in the
study FFQ at baseline, which likely influenced the results. Some patients with high plasma PL
marine n-3FA levels reported only average marine n-3FA intake according to the study FFQ, all
of whom reported frequent use of cod liver oil. Discussion This signals that the study FFQ weighted
response scale for cod liver oil likely underestimated marine n-3FA content, thus the study FFQ
in its current form lacks precision for patients taking daily marine n-3FA supplements. Addi-
tionally, patients with average plasma PL marine n-3 FA levels who reported high levels of
marine n-3FA intake in the FFQ, showed this pattern both at baseline and end of study. This
might be due to social desirability bias and has likely influenced results at both time-points. Fish intake in Norway is higher than in most other European countries, due to the rich fish-
ing grounds along the Norwegian coastline with easy access to fresh cold-water fish [36]. Plasma PL marine n-3FA levels in the present cohort were relatively high, even for a Norwe-
gian population, signaling a selected population that focuses on healthy eating habits. On the
other hand, plasma PL marine n-3FA levels in the present study were comparable to a previous
large cohort study in Norwegian RTRs, suggesting that the sample was representative of a Nor-
wegian transplant cohort [3]. Confounding factors like socioeconomical class, educational
level, smoking habits and physical activity may influence associations between fish intake and
outcomes in epidemiological research [36]. In this cohort, FFQ based marine n-3FA consump-
tion estimates and plasma PL marine n-3FA level were associated with smoking habits, but not
other life-style factors. Dietary habits are changing in the Nordic countries, with lower fish consumption in youn-
ger patients, including Norwegian RTRs [3], thus necessitating revision of questions and
response categories for the present study FFQ in future studies. Cod liver oil intake is an old
tradition in Norway [37] and was therefore included as one of the food times in the study FFQ. This may be omitted in areas where intake of cod liver oil or other marine n-3FA supplements
are uncommon. Strengths of the present study include a well-described cohort, plasma PL fatty acid analysis
and a study FFQ performed at two time points, which might improve accuracy. The study
FFQ has a simple design, is easy to read and understand and only takes a few minutes to
answer, which is desirable in large epidemiological studies. There were also several limitations, including limitations by design such as recall bias and
social desirability bias and a relatively small sample size. Discussion The study FFQ marine n-3FA con-
sumption estimates were based on the sum of weighted response scales for ten food items, con-
taining questions on how frequent the food items were consumed, but not on portion size. Thus, the weighted responses used to calculate marine n-3FA intake were based on assump-
tions of standard portion size for each item, constituting a major limitation in the present
study. Moreover, the study FFQ did not contain any questions regarding seasonal variations,
which could be relevant for some of the included food items in a Norwegian cohort. The study
FFQ contains rather detailed questions about fish and seafood intake and response categories
with minor differences (Fig 1). This likely improved precision for patients who are well aware
of their eating habits but could have been challenging for other patients, possibly leading to
random responses. Broader response categories might have produced more reliable data [38]. For patients on marine n-3FA supplements, like cod liver oil, weighted responses for this food
item in the study FFQ likely underestimated supplements as a source of marine n-3FAs. The questionnaire was designed to estimate marine n-3FA consumption in a Norwegian
transplant cohort. Due to dietary differences between regions and between patient popula-
tions, FFQ validation studies designed for one region or one particular target population may
not apply to other regions or other patient cohorts [22]. In other regions, food items and
weighted response scales should be revised to reflect fish consumption in that region. PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 9 / 14 PLOS ONE Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort Moreover, adjustment for portion size and seasonal variations can be made to improve FFQ
performance. In conclusion, marine n-3FA consumption estimates based on our study FFQ showed a
moderate correlation with the reference biomarker plasma PL marine n-3FA levels, with com-
parable performance to previous FFQs. We recommend using fatty acid analysis to ensure
objective measurement of marine n-3FA consumption in clinical trials, but our FFQ might be
useful in epidemiological studies where resources are limited. Supporting information S1 Fig. Study food frequency questionnaire focusing on food items containing marine n-3
fatty acids (Norwegian version). The study subjects responded to the question “During a typi-
cal month, how often do you eat these food items?” using one of six response alternatives for
each of the ten food item categories. (PDF) S2 Fig. Study food frequency questionnaire focusing on food items containing marine n-3
fatty acids (investigator’s scoring sheet version in English). The study subjects responded to
the question “During a typical month, how often do you eat these food items?” using one of six
response alternatives for each of the ten food item categories. Based on EPA and DHA content
in the meat of various fish and other seafoods presented in the US Department of Agriculture
Food Composition Database and assuming a standard portion size for dinner and bread
spread, every potential response was given a weight (shown inside boxes). Total intake of
marine n-3 fatty acids per month was calculated as the sum of the ten weighted responses in
grams. S3 Fig. Study food frequency questionnaire focusing on food items containing marine n-3
fatty acids (investigator’s scoring sheet version in English comprising fatty fish items
only). The study subjects responded to the question “During a typical month, how often do
you eat these food items?” using one out of six response alternatives for each food item catego-
ries. Based on EPA and DHA content in the meat of various fish and other seafoods presented
in the US Department of Agriculture Food Composition Database and assuming a standard
portion size for dinner and bread spread, every potential response was given a weight (shown
inside boxes). Total intake of marine n-3 fatty acids per month was calculated as the sum of
the weighted responses in grams, which for fatty fish intake comprised the four items shown. (PDF) S4 Fig. Study food frequency questionnaire focusing on food items containing marine n-3
fatty acids (investigator’s scoring sheet version in English comprising fish servings per
month). The study subjects responded to the question “During a typical month, how often do
you eat these food items?” using one out of six response alternatives for each of the ten food
item categories. Servings of fish per month was calculated as the sum of the ten responses,
using the center value for each response category as shown. (PDF) S5 Fig. Author Contributions Conceptualization: My Svensson, Trond Jenssen, Erik B. Schmidt, Ivar A. Eide. Data curation: Ivar A. Eide. Formal analysis: Joe Chan, Ivar A. Eide. Funding acquisition: My Svensson. Funding acquisition: My Svensson. Investigation: Erik B. Schmidt, Ivar A. Eide. Investigation: Erik B. Schmidt, Ivar A. Eide. Methodology: Joe Chan, My Svensson, Trond Jenssen, Erik B. Schmidt, Ivar A. Eide. Methodology: Joe Chan, My Svensson, Trond Jenssen, Erik B. Schmidt, Ivar A. Eide. Project administration: Trond Jenssen, Ivar A. Eide. Resources: My Svensson, Trond Jenssen, Erik B. Schmidt, Ivar A. Eide. Supervision: My Svensson, Erik B. Schmidt, Ivar A. Eide. Supervision: My Svensson, Erik B. Schmidt, Ivar A. Eide. Validation: Ivar A. Eide. Validation: Ivar A. Eide. Writing – original draft: Joe Chan. Writing – original draft: Joe Chan. Writing – review & editing: Joe Chan, My Svensson, Trond Jenssen, Erik B. Schmidt, Ivar A. Eide. Supporting information Scatterplots of standardized plasma marine n-3FA levels and standardized FFQ
based marine n-3FA consumption estimates with regression lines at baseline eight weeks
post-transplant for patients belonging to the control group of the ORENTRA trial
(n = 66). (TIF) 10 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 PLOS ONE Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort S6 Fig. Bland-Altman plot assessing degree of agreement between standardized plasma
marine n-3FA levels and standardized FFQ based marine n-3FA consumption estimates at
baseline eight weeks post-transplant for patients belonging to the control group of the
ORENTRA trial (n = 66). (TIF) S7 Fig. Correlation matrix presenting Spearman’s correlation coefficients at eight weeks
(baseline visit) after renal transplantation for the whole study population (n = 132) and
one year post-transplant (end of study visit) for patients belonging to the control group of
the ORENTRA trial (n = 64). (TIF) S1 File. Supporting information. Includes information regarding “Patient screening and
recruitment in the ORENTRA trial”, “Fatty acid analysis”, “Sample Size and Power Calcula-
tion” and “Development of the study Food Frequency Questionnaire”. (DOCX) Acknowledgments We thank coworkers Rikke Bu¨low Eschen, Annette Andreassen, Birthe H. Thomsen and Inge Aar-
destrup at The Lipid Research Laboratory, Aalborg University Hospital, Denmark for analyzing
plasma phospholipid fatty acids. We thank statistician Owen Thomas and colleague dr. Anupam
Chandra at Akershus University Hospital for their contribution to this manuscript. We thank the
funding sources Gidske and Peter Jacob Sørensen Research Fund and the South-Eastern Norway
Regional Health Authority. Finally, we thank the study participants in the ORENTRA trial. References 1. Lim AK, Manley KJ, Roberts MA, Fraenkel MB. Fish oil for kidney transplant recipients. The Cochrane
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outcomes in patients with dysglycemia. N Engl J Med. 2012; 367(4):309–18. Epub 2012/06/13. https://
doi.org/10.1056/NEJMoa1203859 PMID: 22686415. 11. Bork CS, Mortensen LT, Hjelmgaard K, Schmidt EB. Marine n-3 fatty acids and CVD: new insights from
recent follow-up studies and clinical supplementation trials. Proc Nutr Soc. 2020:1–7. Epub 2020/04/
03. https://doi.org/10.1017/S0029665120006886 PMID: 32234084. 12. Yokoyama M, Origasa H, Matsuzaki M, Matsuzawa Y, Saito Y, Ishikawa Y, et al. Effects of eicosapenta-
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label, blinded endpoint analysis. Lancet. 2007; 369(9567):1090–8. https://doi.org/10.1016/S0140-6736
(07)60527-3 PMID: 17398308. 13. Galan P, Kesse-Guyot E, Czernichow S, Briancon S, Blacher J, Hercberg S, et al. Effects of B vitamins
and omega 3 fatty acids on cardiovascular diseases: a randomised placebo controlled trial. BMJ. 2010;
341:c6273. Epub 2010/12/01. https://doi.org/10.1136/bmj.c6273 PMID: 21115589; PubMed Central
PMCID: PMC2993045. 14. Rauch B, Schiele R, Schneider S, Diller F, Victor N, Gohlke H, et al. OMEGA, a randomized, placebo-
controlled trial to test the effect of highly purified omega-3 fatty acids on top of modern guideline-
adjusted therapy after myocardial infarction. Circulation. 2010; 122(21):2152–9. Epub 2010/11/10. https://doi.org/10.1161/CIRCULATIONAHA.110.948562 PMID: 21060071. 15. Bowman L, Mafham M, Wallendszus K, Stevens W, Buck G, Barton J, et al. Effects of n-3 Fatty Acid
Supplements in Diabetes Mellitus. N Engl J Med. 2018; 379(16):1540–50. Epub 2018/08/28. https://doi. org/10.1056/NEJMoa1804989 PMID: 30146932. 16. Manson JE, Cook NR, Lee IM, Christen W, Bassuk SS, Mora S, et al. Marine n-3 Fatty Acids and Pre-
vention of Cardiovascular Disease and Cancer. N Engl J Med. 2019; 380(1):23–32. Epub 2018/11/13. https://doi.org/10.1056/NEJMoa1811403 PMID: 30415637; PubMed Central PMCID: PMC6392053. 17. Bhatt DL, Steg PG, Miller M, Brinton EA, Jacobson TA, Ketchum SB, et al. Cardiovascular Risk Reduc-
tion with Icosapent Ethyl for Hypertriglyceridemia. N Engl J Med. 2019; 380(1):11–22. Epub 2018/11/
13. https://doi.org/10.1056/NEJMoa1812792 PMID: 30415628. 18. Hu Y, Hu FB, Manson JE. Marine Omega-3 Supplementation and Cardiovascular Disease: An Updated
Meta-Analysis of 13 Randomized Controlled Trials Involving 127 477 Participants. J Am Heart Assoc. 12 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 PLOS ONE Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort 2019; 8(19):e013543. Epub 2019/10/01. https://doi.org/10.1161/JAHA.119.013543 PMID: 31567003;
PubMed Central PMCID: PMC6806028. 2019; 8(19):e013543. Epub 2019/10/01. https://doi.org/10.1161/JAHA.119.013543 PMID: 31567003;
PubMed Central PMCID: PMC6806028. 19. 19. References Validation of a Dietary Question-
naire to Screen Omega-3 Fatty Acids Levels in Healthy Adults. Nutrients. 2019; 11(7). Epub 2019/07/
03. https://doi.org/10.3390/nu11071470 PMID: 31261632; PubMed Central PMCID: PMC6682879. 29. Schumacher TL, Burrows TL, Rollo ME, Wood LG, Callister R, Collins CE. Comparison of fatty acid
intakes assessed by a cardiovascular-specific food frequency questionnaire with red blood cell mem-
brane fatty acids in hyperlipidaemic Australian adults: a validation study. European journal of clinical
nutrition. 2016; 70(12):1433–8. Epub 2016/08/11. https://doi.org/10.1038/ejcn.2016.144 PMID:
27507074. 30. Allaire J, Moreel X, Labonte ME, Leger C, Caron A, Julien P, et al. Validation of the omega-3 fatty acid
intake measured by a web-based food frequency questionnaire against omega-3 fatty acids in red
blood cells in men with prostate cancer. European journal of clinical nutrition. 2015; 69(9):1004–8. Epub
2015/03/12. https://doi.org/10.1038/ejcn.2015.7 PMID: 25758837. 31. Brunvoll SH, Thune I, Frydenberg H, Flote VG, Bertheussen GF, Schlichting E, et al. Validation of
repeated self-reported n-3 PUFA intake using serum phospholipid fatty acids as a biomarker in breast
cancer patients during treatment. Nutr J. 2018; 17(1):94. Epub 2018/10/20. https://doi.org/10.1186/
s12937-018-0402-6 PMID: 30333016; PubMed Central PMCID: PMC6192340. 32. Lepsch J, Vaz JS, Moreira JD, Pinto TJ, Soares-Mota M, Kac G. Food frequency questionnaire as an
indicator of the serum composition of essential n-3 and n-6 polyunsaturated fatty acids in early preg-
nancy, according to body mass index. J Hum Nutr Diet. 2015; 28(1):85–94. Epub 2014/03/14. https://
doi.org/10.1111/jhn.12225 PMID: 24620790. 33. Zhou YB, Li HT, Trasande L, Wang LL, Zhang YL, Si KY, et al. A Correlation Study of DHA Intake Esti-
mated by a FFQ and Concentrations in Plasma and Erythrocytes in Mid- and Late Pregnancy. Nutrients. 2017; 9(11). Epub 2017/11/17. https://doi.org/10.3390/nu9111256 PMID: 29144430; PubMed Central
PMCID: PMC5707728. 34. Kobayashi M, Jwa SC, Ogawa K, Morisaki N, Fujiwara T. Validity of a food frequency questionnaire to
estimate long-chain polyunsaturated fatty acid intake among Japanese women in early and late 13 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 PLOS ONE Marine n-3 fatty acid consumption in a Norwegian renal transplant cohort pregnancy. J Epidemiol. 2017; 27(1):30–5. Epub 2017/01/31. https://doi.org/10.1016/j.je.2016.07.001
PMID: 28135195; PubMed Central PMCID: PMC5328737. pregnancy. J Epidemiol. 2017; 27(1):30–5. Epub 2017/01/31. https://doi.org/10.1016/j.je.2016.07.001
PMID: 28135195; PubMed Central PMCID: PMC5328737. 35. Liu MJ, Li HT, Yu LX, Xu GS, Ge H, Wang LL, et al. PLOS ONE | https://doi.org/10.1371/journal.pone.0244089
December 17, 2020 38.
Cade J, Thompson R, Burley V, Warm D. Development, validation and utilisation of food-frequency
questionnaires—a review. Public Health Nutr. 2002; 5(4):567–87. Epub 2002/08/21. https://doi.org/10.
1079/PHN2001318 PMID: 12186666. References A Correlation Study of DHA Dietary Intake and
Plasma, Erythrocyte and Breast Milk DHA Concentrations in Lactating Women from Coastland, Lake-
land, and Inland Areas of China. Nutrients. 2016; 8(5). Epub 2016/05/24. https://doi.org/10.3390/
nu8050312 PMID: 27213448; PubMed Central PMCID: PMC4882724. 35. Liu MJ, Li HT, Yu LX, Xu GS, Ge H, Wang LL, et al. A Correlation Study of DHA Dietary Intake and
Plasma, Erythrocyte and Breast Milk DHA Concentrations in Lactating Women from Coastland, Lake-
land, and Inland Areas of China. Nutrients. 2016; 8(5). Epub 2016/05/24. https://doi.org/10.3390/
nu8050312 PMID: 27213448; PubMed Central PMCID: PMC4882724. 36. Micha R, Khatibzadeh S, Shi P, Fahimi S, Lim S, Andrews KG, et al. Global, regional, and national con-
sumption levels of dietary fats and oils in 1990 and 2010: a systematic analysis including 266 country-
specific nutrition surveys. BMJ. 2014; 348:g2272. Epub 2014/04/17. https://doi.org/10.1136/bmj.g2272
PMID: 24736206; PubMed Central PMCID: PMC3987052. 37. Mai XM, Langhammer A, Chen Y, Camargo CA Jr., Cod liver oil intake and incidence of asthma in Nor-
wegian adults—the HUNT study. Thorax. 2013; 68(1):25–30. Epub 2012/09/15. https://doi.org/10. 1136/thoraxjnl-2012-202061 PMID: 22977130. 37. Mai XM, Langhammer A, Chen Y, Camargo CA Jr., Cod liver oil intake and incidence of asthma in Nor-
wegian adults—the HUNT study. Thorax. 2013; 68(1):25–30. Epub 2012/09/15. https://doi.org/10. 1136/thoraxjnl-2012-202061 PMID: 22977130. 38. Cade J, Thompson R, Burley V, Warm D. Development, validation and utilisation of food-frequency
questionnaires—a review. Public Health Nutr. 2002; 5(4):567–87. Epub 2002/08/21. https://doi.org/10. 1079/PHN2001318 PMID: 12186666. 38. Cade J, Thompson R, Burley V, Warm D. Development, validation and utilisation of food-frequency
questionnaires—a review. Public Health Nutr. 2002; 5(4):567–87. Epub 2002/08/21. https://doi.org/10. 1079/PHN2001318 PMID: 12186666. 14 / 14
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55 Ibidem. Legajo, f. 61v.
56 Ibidem. Legajo, f. 62r.
140
AEA, 66, 2, julio-diciembre, 2009, 125-145. ISSN: 0210-5810
EL CASO DE MATEO CHIMALTECUHTLI (CHOLULA, SIGLO XVI)
El teniente decidió no conceder a Mateo lo que pedía y, además, afirmó que este no sacaba el proceso por malicia. El 26 de enero de 1565,
Isabel presentó una petición para que se cumpliese la sentencia. Ante esta
y la “malicia” de Mateo, el corregidor decidió dar cumplimiento a esta y
prender a Mateo hasta que así fuese. No tenemos constancia en el documento de que eso se ejecutase. Sin embargo, sí aparece un mandamiento de
la Real Audiencia en el que se indica:
que En la n[uest]ra | avdiençia corte E chançilleria que Reside En la çibdad | de mexico de la nueva espana antel preSidente E | oydores della paresçio mateo chimalteCatl
yndio | natural de la çibdad de chulula E se preSento con | vna petiçion En grado de
apelaçion nulidad | E agravio de vna sentençia contra el dada E | pronunçiada por el
corregidor de la d[ic]ha çibdad y En | fabor de x[hris]poval su hermano que nos pedia
E Su|plicaua le oviesemos por preSentado En el d[ic]ho grado | E darle n[uest]ra
prouiSion conpulsoria para que le | dieSedes Vn traslado del proçeso E abtos sobre
Ello | fechos o que sobre Ello proveyeSemos como la | n[uest]ra m[erçe]d fuese lo
qual por los d[ic]hos n[uest]ro presidente | E oydores Visto fue acordado que deviamos | mandar dar Esta n[uest]ra carta Esta n[uest]ra carta [sic.] En | la d[ic]ha Razon
E nos tovimos lo por bien por la qual | Vos mandamos que dentro de quatro dias primeros | Siguientes de como con ella fueredes Requeridos | deis y Entregueis a la parte del d[ic]ho mateo chimal | vn traslado del proçeso del d[ic]ho pleito que de suso se
ha|ze minçion con todos E qualesqu(i)er ab(t)os a el tocantes | E pertenesçientes
EsCritos En limpio conforme | al aranzel destos n[uest]ros Reinos f[i]rmado Signado
| çeRado E sellado En publica forma En | manera que haga fee pagando os los
d[ere]c[h]os que por ello | huvieredes de aver los quales aSentad E f[i]rmad | al pie
dello para que lo pueda traer E pre[se]ntar | ante los d[ic]hos n[uest]ro presidente E
oydores | para guarda de su d[ere]c[h]o y non faga desEnde al por | alguna manera So
pena de la n[uest]ra m[erçe]d E de cient | pesos de oro para la n[uest]ra camara dada
En la çibdad | de mexico A catorze dias del mes de abrill de mill E | quie[nient]os E
sesenta E çinco anos El doctor çey|nos El doctor VillaloVos El doctor Villanue|va yo
gordian casasano EsCriuano de Cama|ra y delavdiençia E chançilleria Real de la
nue|va espana por su mag[es]t[ad] la fize EsCriuir por su | mandado con aCuerdo de
su presydente E oydores | Registrada Juan SeRano chançiller Juan orgus |
t[esti]m[onio] ||.57
Con dicho mandamiento, Mateo logró finalmente obtener una copia
del proceso para recurrir la sentencia, que es lo que actualmente se conserva. 57 Ibidem. Legajo, f. 64r y v.
AEA, 66, 2, julio-diciembre, 2009, 125-145. ISSN: 0210-5810
141
MIGUEL ÁNGEL RUZ BARRIO
Por la documentación que conocemos, sólo podemos afirmar que
Mateo parece que inició los trámites para llevar a cabo su apelación. Sin
embargo no sabemos cuál fue el resultado. No se ha localizado ningún
documento que atestigüe que llegase a la Audiencia y por tanto desconocemos si esta falló a favor o en contra. Lo que sí tenemos es el que consideramos como testamento de Mateo Chimaltecuhtli, la Memoria de don
Matheo Caxco,58 en el que se mencionan diversos objetos que aparecen en
el litigio. Por ello, podemos suponer que de una u otra manera
Chimaltecuhtli consiguió eludir la sentencia desfavorable y conservar las
propiedades.
Comentario del pleito
Todo este documento nos está reflejando parte de la práctica jurídica
de la administración española frente a los pleitos entre indígenas, pero también la actitud de estos ante ella. No sabemos si esta causa se presentó ante
las autoridades indígenas, aunque tal vez, al no mencionarse nada, debemos entender que no. Por tanto el litigio se inicia con la acción de demanda puesta por Isabel. Ella era el actor que presenta como pruebas iniciales
la pintura y su propio testimonio para apoyar la causa. Sin embargo, en
varios momentos se menciona su ignorancia respecto a la Justicia, y así,
por ejemplo, cuando pone la demanda afirma lo siguiente:
En muy | gran perJuizio dela susod[ic]ha y del d[ic]ho pablo | chimalteCutli porques
heredero ligitimo delo | susod[ic]ho E como la susod[ic]ha Es prove E yno|rante de
pedir su Justiçia y el d[ic]ho pablo su hijo | Es menos de diez E seis anos E ques
çiego | E persona muy sinple E que no an te|nido abilidad para pedir su Justiçia hasta a|gora quela susod[ic]ha y el d[ic]ho su hijo se an visto E | se been muy pobres E
nesçesitados E por lo | que toca a su conçiençia pone Esta deman|da al d[ic]ho mateo
para que no posea lo ques del | d[ic]ho pablo su hijo pueslo tiene E posee
yn|Justamente.59
No podemos determinar si se trataba de una realidad o de una postura para justificar por qué no había solicitado antes justicia. A pesar de todo
el corregidor valoró la causa como justificada y procedió a notificarle la
demanda a Mateo.
58 Ibidem, pp. 346-349.
59 Ibidem. Legajo, f. 42v.
142
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EL CASO DE MATEO CHIMALTECUHTLI (CHOLULA, SIGLO XVI)
La actitud de Chimaltecuhtli parece muy distinta a la de Isabel frente
a la Justicia. Aunque indica en varios momentos que no sabe escribir, se
presenta con un escrito náhuatl en alfabeto europeo. Por tanto, parece que
se mueve en un círculo distinto al de Isabel, quien había presentado una
pintura. El valor de ambos documentos debió ser similar,60 pero sin embargo creemos que Mateo da muestras de una mayor adaptación a las novedades. Esto se verá acentuado más adelante, ya que, a pesar de que parece que
no consiguió probar bien su postura, Mateo se las ingenió para retrasar el
cumplimiento de la sentencia. En primer lugar, presentó la petición por
escrito, en castellano, de apelar ante la Real Audiencia, que el corregidor le
concedió. En segundo, tras esta concesión, solicitó, de nuevo, por escrito
en castellano, un traslado del expediente del pleito, para mostrar a la persona que le ayudaría en la apelación y solicitaba: “se me de y termino de
abo/gado”.61 Ante ello el corregidor comenzó a considerar que Mateo
actuaba con malicia y no se lo otorgó.
De nuevo Mateo presentó un escrito más en español reclamando un
traslado del pleito. En este caso señalaba que lo pretendía llevar a Puebla,
donde Diego de Baeza lo vería para ayudarle.62 Aquí tenemos la persona
que tal vez asesoraba desde un principio a Mateo y podemos suponer que
se trataba de un abogado. Además, Mateo debía tener contacto con alguien
que sabía escribir en alfabeto europeo, mientras que es probable que Isabel
no. Ella sólo presentó una pintura y tal vez sin glosas en alfabeto europeo.63
Las personas que escribieron las cartas de Mateo pudieron ser varias, ya
que aparecen en náhuatl y en castellano, pero también pudo hacerlo una
sola o incluso él mismo.
No sabemos realmente que ocurrió entonces, aunque lo más probable es que todo sucediese tal y como lo mandaba el corregidor. La
siguiente fecha que tenemos es de abril, cuando Mateo presentó un escrito de la Real Audiencia en el que se le daba permiso para la apelación y
para sacar el traslado. Por tanto, fue el escribano, Francisco Muñoz, quien
recibió la petición de Mateo para cumplir con la orden de sacar el traslado y así lo hizo. De este modo termina la documentación que tenemos
sobre el proceso, en el que da la impresión de que ambas partes hacen uso
60 Véase Ruz Barrio: “Los códices jurídicos: definición y metodología de estudio”. Desacatos.
Revista de Antropología Social, México, en prensa.
61 Ruz Barrio: Un conjunto de documentos inéditos…, Legajo, f. 61v.
62 Ibidem, Legajo, f. 62 r.
63 Ibidem, pp. 273-322.
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143
MIGUEL ÁNGEL RUZ BARRIO
de la Justicia española con bastante solvencia. Sin embargo, parece que
Mateo era más experto o estaba mejor asesorado y trataba de conseguir
que se dilatase cada vez más el proceso, tal vez debido a que fue él quien
perdió.
Lo cierto es que ambas partes demuestran tener ciertas nociones del
funcionamiento del sistema. En los dos casos, es muy probable que hubiesen tenido a alguien que los asesorase, aunque no aparece claramente en el
expediente. Para Mateo sí conocemos un nombre, Diego de Baeza, quien le
ayudaría en su apelación. Este personaje reside en Puebla. Además Mateo
consigue el permiso de la Real Audiencia; por tanto se movía, ayudado o
no, con cierta solvencia en el sistema.
Como vimos en nuestra visión general sobre los pleitos indígenas,
este es un claro ejemplo donde la idea de los juicios rápidos concebidos por
la administración choca con los intereses de una de las partes, que ante la
sentencia desfavorable hace uso de toda la maquinaria legal. Aunque el
corregidor dio mandato de posesión a Isabel y su hijo en enero y la carta de
la Audiencia parece ser de abril, no sabemos si se llegó a producir de manera efectiva. Por tanto estaríamos ante un caso donde el dilatar el proceso no
era del todo perjudicial para los indígenas, al menos para una de las partes.
Tal vez para Mateo esto suponía mantener la posesión, haciendo así menos
graves los costos del proceso. Además, si, como se dice en el pleito, Isabel
se encontraba pobre, es probable que no pudiese mantener ese largo proceso y eso le diese el triunfo a Mateo. Por el momento, no conocemos ninguna referencia sobre cómo terminó todo. Sabemos que Francisco Muñoz
sacó el traslado y que probablemente fue él quien puso el texto del f. 39r
donde dice:
[Cruz] ||
[Calderón] | proçeso de demanda de ysabel Eçi y mateo | chimaltecutli yndios de
la çiudad de chulula sobre | vnas tierras y Joias y ua En grado de app[elaci]on | hecha
por el d[ic]ho mateo a la Real audiençia (des)|ta nueva espana çeRado y sellado
[Rúbrica] (Legajo, f. 39r.).
El contenido de este título escrito en la cubierta del pleito (Legajo, f.
39r) hace claramente referencia al documento, que iba cerrado y sellado,
suponemos que con el fin de garantizar que no hubiese añadidos o pérdidas
en él. Sin embargo, faltan elementos que nos indiquen que fue presentado
en esa apelación. A pesar de todo, podemos considerar que de alguna
manera Mateo consiguió burlar la sentencia en su contra y permanecer con
144
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EL CASO DE MATEO CHIMALTECUHTLI (CHOLULA, SIGLO XVI)
la posesión de los objetos en litigio, gracias a la presencia de estos en su
testamento.64
Conclusión
Para cerrar este artículo, únicamente queremos señalar que este pleito
nos ilustra cómo en ocasiones los propios indígenas, o al menos algunos,
son los más interesados en dilatar el desarrollo de los litigios, ya que en ello
les podía ir su triunfo. El caso de Mateo no es una excepción, sino una
muestra de la complejidad del contexto novohispano.
Recibido el 14 de octubre de 2008
Aceptado el 7 de enero de 2009
64 Ibidem., pp. 346-349.
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145.
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OA04.02. Mechanisms of growth inhibition of pancreatic cancer by omega-3 polyunsaturated fatty acids
|
M Chen
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English
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Purpose Omega-3 polyunsaturated fatty acids (PUFAs) are widely
considered health promoting. We have previously
reported that the omega-3 PUFA eicosapentaenoic acid
(EPA) inhibits growth of pancreatic cancer (PaCa) cells
in vitro and in vivo. However, the mechanism underly-
ing the effects of EPA in PaCa cells is unknown. OA04.02. Mechanisms of growth inhibition of
pancreatic cancer by omega-3 polyunsaturated
fatty acids From International Research Congress on Integrative Medicine and Health 2012
Portland, Oregon, USA. 15-18 May 2012 phosphorylation and increased FASN expression, while
LY294002 reduced FASN. PaCa cells transfected with
myr-Akt1 exhibited increased FASN expression, indicating
the importance of Akt in baseline FASN expression. Importantly, EPA decreased Akt phosphorylation and
FASN expression in control-transfected cells, but this inhi-
bition was abrogated in myr-Akt1 transfected cells. phosphorylation and increased FASN expression, while
LY294002 reduced FASN. PaCa cells transfected with
myr-Akt1 exhibited increased FASN expression, indicating
the importance of Akt in baseline FASN expression. Importantly, EPA decreased Akt phosphorylation and
FASN expression in control-transfected cells, but this inhi-
bition was abrogated in myr-Akt1 transfected cells. Results EPA dose-dependently increased apoptosis and stimulated
cleavage of PARP and caspase-3/7, which was accompa-
nied by inhibition of Akt. Exposure of PaCa cells to
LY294002 induced apoptosis and decreased cell growth. EPA-induced apoptosis and growth inhibition was attenu-
ated in myr-Akt1 transfected cells. FASN was expressed in
all cell lines. Inhibition of FASN by C75, a synthetic FASN
inhibitor, reduced cell growth and induced apoptosis. EPA
inhibited FASN expression, which was accompanied by
inhibition of cell growth. Insulin stimulated Akt Conclusion Six human PaCa cell lines of varying degrees of differen-
tiation were exposed to different concentrations and
times to EPA. Cell growth was measured by BrdU and
apoptosis was determined by a Cell Death ELISA, and
cleavage of PARP and caspase-3/7. The involvement of
PI3K/Akt, a major survival pathway in PaCa, was deter-
mined using pharmacological inhibitors (LY294002) and
constitutively active Akt (myr-Akt1). The effect of EPA
on de novo fatty acid synthesis as a downstream pathway
of Akt inhibition was investigated using Western blot-
ting and inhibitors of fatty acid synthase (FASN). Our studies provide compelling evidence that the
growth inhibitory effects of the omega-3 polyunsatu-
rated fatty acid EPA in PaCa cells was mediated by inhi-
bition of the PI3K/Akt pathway and subsequently
reduced fatty acid synthesis. Published: 12 June 2012 Published: 12 June 2012 doi:10.1186/1472-6882-12-S1-O14
Cite this article as: Chen et al.: OA04.02. Mechanisms of growth
inhibition of pancreatic cancer by omega-3 polyunsaturated fatty acids. BMC Complementary and Alternative Medicine 2012 12(Suppl 1):O14. doi:10.1186/1472-6882-12-S1-O14
Cite this article as: Chen et al.: OA04.02. Mechanisms of growth
inhibition of pancreatic cancer by omega-3 polyunsaturated fatty acids. BMC Complementary and Alternative Medicine 2012 12(Suppl 1):O14. Chen et al. BMC Complementary and Alternative Medicine 2012, 12(Suppl 1):O14
http://www.biomedcentral.com/1472-6882/12/S1/O14 Chen et al. BMC Complementary and Alternative Medicine 2012, 12(Suppl 1):O14
http://www.biomedcentral.com/1472-6882/12/S1/O14 Chen et al. BMC Complementary and Alternative Medicine 2012, 12(Suppl 1):O14
http://www.biomedcentral.com/1472-6882/12/S1/O14 Open Access David Geffen School of Medicine at UCLA, Department of Surgery, Los
Angeles, USA © 2012 Chen et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons
Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in
any medium, provided the original work is properly cited. Submit your next manuscript to BioMed Central
and take full advantage of: • Convenient online submission David Geffen School of Medicine at UCLA, Department of Surgery, Los
Angeles, USA © 2012 Chen et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons
Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in
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| 31,148
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https://openalex.org/W2329686972
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OpenAlex
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Open Science
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Public Domain
| 1,913
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Vegetation of New Mexico
|
John Watson
|
English
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Spoken
| 2,999
| 4,655
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Relation of Prairie and Forest in Central Illinois 155 improbable that the whole surface was ever forested; the level plains between the stream systems
and the moraines were probably prairie even at the time of greatest forest advance, and the
immigration of the forest as restricted to the two lines of greatest physiographical diversity-the
stream valleys and the moraines. The reason for the removal of this large body of forest from the moraine and for the per-
sistence of the small remainder in a few outlying tracts like that of Bur Oak Grove is suggested
by examination of the conditions in the grove and by other observations elsewhere in the area. Along the western margin of the grove some of the ridges are still forested, while others are
under cultivation. From the presence of typically forest plants (Aster Drlcummondii, Silene
stellata, etc.) in the vegetation along the roadsides on the cultivated ridges, it is evident that
these ridges were originally forested also. On some other ridges these species are absent, the
roadside vegetation consisting of typically prairie species of Andropogon, Panicuvm, Silphinm,
Partheniurn, etc.-evidently
these ridges were originally prairie. It thus becomes evident, the
exact boundaries of the grove being determined by observation of the relic plants, that in every
case those ridges which are or were forested are protected on the west by a conspicuous slough,
while the prairie ridges extend west without interruption out upon the open prairie. Since the
forested part of the grove is exclusively on the ridges, it is clear that the whole forest was
protected on the western side by a series of sloughs. The prevailing winds are also from the
west, and prairie fires driven eastward by a west wind were unable to cross the slough into
the forest. Hence it may be concluded that prairie fires were the chief and probably the only
agent in the removal of the forest from the moraines and other places where it was not properly
protected by a water barrier. The grove at Bur Oak was favoured by a peculiar and unusual
topography and was virtually the only portion of an extensive forest system to be spared. p g
p y
y
y p
y
p
The origin of the prairie as a type of vegetation cannot, however, be referred to prairie fires as
a cause, as was frequently supposed by early writers and occasionally even in recent years. Relation of Prairie and Forest in Central Illinois A
prairie fire presupposes a prairie, and in prairie fires we have merely one factor which has played
a part in the maintenance or extension of the prairie in its struggle against forest invasion. In
the last half century, since the cessation of prairie fires, the forests have again begun an advance
into the prairie, but their route is chiefly up the streams, and the migration is limited to a
comparatively small number of mobile species. Owing to increasing cultivation this migration is
very irregular and can never lead to any serious modification in the vegetation of the region. The conditions in the grove studied serve to indicate the last three periods in the vegetational
history of Illinois-(1)
period of forest advance, leading to a great development of forests in areas
of physiographic diversity; (2) period of prairie fires, following the advent of man and leading to
the restriction of the forest to protected areas and the corresponding extension of the prairie;
(3) period of civilisation and the virtual cessation of the struggle between forest and prairie. Review: Vegetation of New Mexico
Source:
Journal of Ecology, Vol. 1, No. 2 (Jun., 1913), pp. 155-157
Published by:
British Ecological Society
Stable URL:
http://www.jstor.org/stable/2255697
Accessed: 13-03-2015 17:08 UTC Review: Vegetation of New Mexico
Source:
Journal of Ecology, Vol. 1, No. 2 (Jun., 1913), pp. 155-157
Published by:
British Ecological Society
Stable URL:
http://www.jstor.org/stable/2255697
Accessed: 13-03-2015 17:08 UTC Your use of the JSTOR archive indicates your acceptance of the Terms & Conditions of Use, available at
http://www.jstor.org/page/info/about/policies/terms.jsp JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content
in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms of scholarship. For more information about JSTOR, please contact support@jstor.org. British Ecological Society is collaborating with JSTOR to digitize, preserve and extend access to Journal of Ecology. http://www.jstor.org This content downloaded from 128.235.251.160 on Fri, 13 Mar 2015 17:08:14 UTC
All use subject to JSTOR Terms and Conditions Relation of Prairie and Forest in Central Illinois This content downloaded from 128.235.251.160 on Fri, 13 Mar 2015 17:08:14 UTC
All use subject to JSTOR Terms and Conditions VEGETATION
OF NEW
MEXICO Watson,
J. R. "Plant
geography of Nortlh Central New Mexico."
Bot. Gazette, 54, 1912, pp. 194-217,
7 text-figures. In the northern half of New Mexico, traversed by the 35th parallel of latitude, which indicates
hot sun in summer and warm in winter, the altitude varies from 5000 feet in the Rio Grande
valley to 11,000 feet in the Sandia mountain range, the topography is diversified, and there is an
abrupt change in vegetation corresponding with the abrupt difference in climate, the arid climate
of the south-west meeting in the mountains the more humid one of the north and east. In the
centre of the region is the valley, two or three miles wide, of the Rio Grande, a shallow muddy
stream which may be half a mile wide during the June melting of the snow on the Colorado
mountains, or entirely dry in August, exposing extensive mud flats on which a vigorous vegetation
rapidly develops. On either side the valley is limited by the much dissected edge of the mesa,
with sand dunes on the western side bearing no vegetation, from which there slopes upward to the
mountains a clinoplain crossed every two or three miles by a sandy " arroyo" (dry stream bed)
which once or twice each summer becomes a raging torrent for a few hours. The most important
climatic factor is aridity, the valley rainfall (at AlbuLquerque) averaging 18 7 cm., that in the Notices of Work on Foreign Vegetation 156 higher mountains probably about 60 cm.; snow, rare in the valley, covers the higher mountains
for a large portion of the winter, and on melting it saturates the soil much more thoroughly than
the summer rains which quickly run off. higher mountains probably about 60 cm.; snow, rare in the valley, covers the higher mountains
for a large portion of the winter, and on melting it saturates the soil much more thoroughly than
the summer rains which quickly run off. Floristically the region is of great interest, being the meeting-place of the northern and eastern
flora with that of the arid south-west-the
genera and most of the species in the mountains ale
identical with those of the east, but those in the mesa are quite different; and a particularly good
one for the study of physiographic plant ecology, because of the abrupt differences in physiography
and climate. Floristically the region is of great interest, being the meeting-place of the northern and eastern
flora with that of the arid south-west-the
genera and most of the species in the mountains ale
identical with those of the east, but those in the mesa are quite different; and a particularly good
one for the study of physiographic plant ecology, because of the abrupt differences in physiography
and climate. The following formations and associations are described. (1) River valley formations, divided
into (a) cottonwood forest, an open and more or less pure Poputluts WTislizeniii forest along the Rio
Grande, with the trees small, including also a few willows, clumps of the shrubs Bacc7laris
JVr'ightii and Cassia bauthiniioides, with ground vegetation of Jutncus balticuts, Trifoliui R1ydbergii,
Aster spi?zosits, and some grass; (b) alternating with (a) in possession of the river banks, a meadow-
like Ju7?cus-Houtttynia
association with J. balticits and H. californica dominant; (c) on the
higher ground a Bigelovia association, dominated by Bigelovia (Chrysothamnus)
Bigelovii, a low
shrubby almost leafless perennial with evergreen shoots, rivalled in dominance in sandy places by
Yuicca glauca, and where the sand is deep and extensive, as in the wider valleys or arroyos, by
Parosela (Dalea) scoparia. The arroyos of the dissected edge of the mesa show an interesting
succession of societies characterised by successively smaller plants as one ascends; these are
described in some detail. Notices of Work on Foreign Vegetation (2) Mesa formation, occupying the more level ground of the mesa and
stopping abruptly at its dissected edge; originally grassland, this has been over-grazed and
invaded by the goldenrod-like Composite Gttierr ezia, and should probably be classified as a steppe,
with monotonously uniform vegetation divisible into three ecological groups-(a)
plants like
Opuntia, Bigelovia, Yuteca, Sarcobatuts, and Staeda, with large underground stems and roots in
which water is stored, (b) annuals and perennials with underground stems, including most of the
mesa herbs and grasses, (c) a few winter annuals like Phacelia corrugata, "loco " weeds (Astragilus
sp.), Draba, Gilia, etc. (3) Cedar formation, with Juniperus monosperm1ba dominant, in the
mountains and the higher parts (6500 feet and over) of the mesa plains; the Gutierrezia and
Yuicca glauca extend into this formation, Opintia arborescens is abundant and characteristic, with
other evergreen and sub-evergreen plants, and there is less difference between the winter and
summer aspects of this formation than of any other because there is less difference in relative
humidity of the soil. (4) Pinon formation, shading into the last very gradually; the pinon (Pimus
edulis) never extends so far down the mountain side as does the cedar. (5) Yellow pine association,
coinciding closely with the region of deep winter snow and marked by a sharp and complete
change of flora-there is much more difference between this formation and the mesa or even the
pinon formation less than a mile away than there is between it and the woods of Ohio or probably
even Europe or Japan; this association descends in places to 7000 feet, and extends to the top of
the range at 10,000 feet, and the dominant Pinus ponderosa is replaced by red cedar (Juniperus
scopldoruni) along lime-charged streams, while in the Sandia Mountains the white oak is charac-
teristic and grows in more xerophytic situations than the pine. In open but less xerophytic
positions on the top of the range the oak chaparral is replaced by mountain meadow composed
not chiefly of grasses but of low herbs (Potentilla, Castilleja, Brickellia, Chrysopsis, Oxytropis,
Achillea, etc.). This content downloaded from 128.235.251.160 on Fri, 13 Mar 2015 17:08:14 UTC
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All use subject to JSTOR Terms and Conditions Notices of Work on Foreign Vegetation Notices of Work on Foreign Vegetation (6) Douglas spruce formation, with Pseudotsutga taxifolia dominant, covering
north-facing slopes above 8000 feet and extending down the narrower cailons to about 7000 feet;
this is the ' Canadian zone " of Merriam, the most mesophytic and dense of the forests in the
region, but replaced on the higher parts by blue spruce (Abies conicolor) and on the highest and
most exposed by Engelmann's spruce (Picea Engelmnan?dii) which answers to Merriam's "Hud-
sonian zone."
(7) Canion associations, a series of ascending successions; the first tree met in
approaching the mountains from an arroyo is the hackberry (Celtis reticutlata), next a society
dominated by box elders-these
seem to be the canion representatives of the cedar and pinon
formations respectively; higher up and in the narrower more mesophytic portions, a society
dominated by Populits anigutstifolia, answering to the yellow pine association; still higher, the
spruce formation holds full sway; and near the head of the can-on, above the permanent stream,
is an association of quaking aspens. The biotic succession in the Sandia mountains is as follows: the bare rock first encrusted with
crustaceous lichens; then foliose lichens, mosses, herbs, oaks, followed in some cases directly by MOUNTAIN
AND
ALPINE
LABORATORIES,
COLO R ADO We have received prospectuses giving particulars of the summer courses of instruction and
investigation undertaken at (1) the Alpine Laboratory of the University of Minnesota, and (2) the
Mountain Laboratory of the University of Colorado. The presence of two " outdoor " laboratories
of this kind in a single State indicates sufficiently the ardour and enterprise of American ecologists,
while a perusal of the particulars given shows how thorough are the methods employed, on one
hand to attract teaching and other botanists to take up ecological work by means of vacation
courses, and on the other to afford facilities for research by more advanced workers. (1) The GTraduate School of Ecology, to be held from July 1 to September 1, at the Alpine
Laboratory, situated at 8500 feet on the cog railway between Manitou and the summit of Pike's
Peak, is to be conducted by Prof. F. E. Clements, assisted by Edith Clemeiits, R. J. Pool, and
H. L. Shantz. The rich and varied flora, with the remarkable diversity of habitat found in this
rugged mountain region, offers exceptional facilities for the study of plant response and the origin
of new forms. Among the alpine summits of the American continent, Pike's Peak is unique in
the series of great formational zones which lies across its face. From the Great Plains grasslands,
the series runs from valley woodland at 5800 feet to mesa, chaparral, foothill woodland, pine
forest, aspen woodland and spruce forest to alpine meadow, rock field and bog at 11,000 to
14,000 feet in a distance of 7 miles. From the very nature of the mountains, weathering, erosion
and other physiographic factors bring about the almost countless repetition of the same or similar
habitats, and produce numbers Qf primary and secondary successions illustrating a wide range of
developmental processes and principles. Ecological work was first done at Pike's Peak in 1899,
and has been carried on each summer since that time; hence it is claimed that probably no other
area has been so intensively studied by means of instrument and quadrat, and offers such a
fascinating array of ecological problems for which the foundation has at least been sketched. The
scope and nature of this foundation work is indicated by the list of publications cited. Vegetation of New Melxico Vegetation of New Melxico 157 Douglas spruce and in others by aspen and then the spruce; and then, as physiographic succession
comes in, the poplars, pines and box elders in the canion, and pine, pinon, and cedar on the
slopes, until the ultimate formation of the mesa is reached. The author points out that the
tendency of the higher zones to creep down the cafions and of the lower zones to creep up the
ridges is more readily explained by reference to the moisture supply in the two regions than to the
cooling and warming effects of descending and ascending currents respectively; and that an
arrangement of " zones " should be based on all factors determining the distribution of life, and
not on one only, especially in a region where that one is of secondary importance. Douglas spruce and in others by aspen and then the spruce; and then, as physiographic succession
comes in, the poplars, pines and box elders in the canion, and pine, pinon, and cedar on the
slopes, until the ultimate formation of the mesa is reached. The author points out that the
tendency of the higher zones to creep down the cafions and of the lower zones to creep up the
ridges is more readily explained by reference to the moisture supply in the two regions than to the
cooling and warming effects of descending and ascending currents respectively; and that an
arrangement of " zones " should be based on all factors determining the distribution of life, and
not on one only, especially in a region where that one is of secondary importance. This content downloaded from 128.235.251.160 on Fri, 13 Mar 2015 17:08:14 UTC
All use subject to JSTOR Terms and Conditions This content downloaded from 128.235.251.160 on Fri, 13 Mar 2015 17:08:14 UTC
All use subject to JSTOR Terms and Conditions This content downloaded from 128.235.251.160 on Fri, 13 Mar 2015 17:08:14 UTC
All use subject to JSTOR Terms and Conditions MOUNTAIN
AND
ALPINE
LABORATORIES,
COLO R ADO The field
for investigation open falls into four general divisions: (i) the use of quantitative methods of
studying habitat and plant; (ii) the application of ecological methods and principles to forestry,
agriculture, and plant pathology; (iii) the measured study of individual response to the habitat
with especial reference to the origin of species; (iv) quadrat study of the development and structure
of plant formations. The practical aspects of quantitative ecology are represented by the Frernont
Forest Experiment Station, and the Dry-land Agriculture Field Station of the U.S. Department
of Agriculture, perhaps the best equipped stations in the world for the exact study of vegetational
problems. (2) The Mountain Laboratory at Tolland, Colorado, was established in 1909, and the work of
the summer session (June 23 to August 2) is conducted by Prof. F. Ramaley. Tolland is a small
hamlet in a mountain valley at 8900 feet altitude. North from the Laboratory are Park Lake
and South Boulder Creek in a broad meadow; to the west are the mountains, rising to the
Continental Divide five miles away; eastward extends the valley of the South Boulder river,
while at the south is a succession of forest-clad hills. The work includes both botany and
zoology, special attention being given to ecology, classification, and field work, with all-day picnic
excursions to alpine heights and to points at various altitudes for the study of plant distribution
as affected by climate. This content downloaded from 128.235.251.160 on Fri, 13 Mar 2015 17:08:14 UTC
All use subject to JSTOR Terms and Conditions
| 41,897
|
https://openalex.org/W2110256701
|
OpenAlex
|
Open Science
|
CC-By
| 2,012
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Human blood RNA stabilization in samples collected and transported for a large biobank
|
Nur Duale
|
English
|
Spoken
| 9,017
| 15,165
|
Abstract Background: The Norwegian Mother and Child Cohort Study (MoBa) is a nation-wide population-based pregnancy
cohort initiated in 1999, comprising more than 108.000 pregnancies recruited between 1999 and 2008. In this study
we evaluated the feasibility of integrating RNA analyses into existing MoBa protocols. We compared two different
blood RNA collection tube systems – the PAXgene™Blood RNA system and the Tempus™Blood RNA system - and
assessed the effects of suboptimal blood volumes in collection tubes and of transportation of blood samples by
standard mail. Endpoints to characterize the samples were RNA quality and yield, and the RNA transcript stability of
selected genes. Findings: High-quality RNA could be extracted from blood samples stabilized with both PAXgene and Tempus
tubes. The RNA yields obtained from the blood samples collected in Tempus tubes were consistently higher than
from PAXgene tubes. Higher RNA yields were obtained from cord blood (3 – 4 times) compared to adult blood
with both types of tubes. Transportation of samples by standard mail had moderate effects on RNA quality and
RNA transcript stability; the overall RNA quality of the transported samples was high. Some unexplained changes in
gene expression were noted, which seemed to correlate with suboptimal blood volumes collected in the tubes. Temperature variations during transportation may also be of some importance. Conclusions: Our results strongly suggest that special collection tubes are necessary for RNA stabilization and they
should be used for establishing new biobanks. We also show that the 50,000 samples collected in the MoBa
biobank provide RNA of high quality and in sufficient amounts to allow gene expression analyses for studying the
association of disease with altered patterns of gene expression. Keywords: Tempus tubes, PAXgene tubes, Cord blood, Quality control, RNA stabilization, Biobank, Blood RNA, Gene
expression Keywords: Tempus tubes, PAXgene tubes, Cord blood, Quality control, RNA stabilization, Biobank, Blood RNA, Gene
expression Norwegian Institute for Public Health (NIPH) together
with extracted DNA, for future use [2]. The DNA is rou-
tinely extracted from the blood samples from each par-
ticipant and frozen in aliquots. In 2005 funds were
obtained from the US National Institutes of Health
through the Autism Birth Cohort to initiate RNA collec-
tion for the purpose of gene expression analyses. Duale et al. BMC Research Notes 2012, 5:510
http://www.biomedcentral.com/1756-0500/5/510 Duale et al. BMC Research Notes 2012, 5:510
http://www.biomedcentral.com/1756-0500/5/510 SHORT REPORT Open Access Human blood RNA stabilization in samples
collected and transported for a large biobank Nur Duale1*, Gunnar Brunborg1, Kjersti S Rønningen1,4, Thomas Briese2, Jeanette Aarem1, Kaja K Aas1, Per Magnus1,
Camilla Stoltenberg1, Ezra Susser2,3 and W Ian Lipkin2 © 2012 Duale et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative
Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited. Abstract The
challenge in implementing RNA collection was to devise
a new protocol that would be feasible in maternity units
and during transportation - and at the same time the
existing routines and logistics of the cohort should be
adhered to as closely as possible. There are several
reports on the ex vivo instability of RNA transcripts
[3-5], and it is therefore very important to stabilize blood
RNA during sample collection, transport, and storage * Correspondence: nur.duale@fhi.no
1The Norwegian Institute of Public Health, Oslo, Norway
Full list of author information is available at the end of the article Background The Norwegian Mother and Child Cohort Study (MoBa)
is a nation-wide population-based pregnancy cohort
initiated in 1999 [1,2]. The MoBa holds more than
108,000 pregnancies recruited between 1999 and 2008;
the last child to be included was born in June 2009. In
addition to detailed questionnaire data, biological mate-
rials from the mother, the father and the child (umbilical
cord blood) in the form of whole blood and plasma, have
been collected and are stored in a biobank at the * Correspondence: nur.duale@fhi.no
1The Norwegian Institute of Public Health, Oslo, Norway
Full list of author information is available at the end of the article Page 2 of 9 Duale et al. BMC Research Notes 2012, 5:510
http://www.biomedcentral.com/1756-0500/5/510 [6], in order to obtain reproducible gene expression
results. establishment of robust and standardized protocols is a
prerequisite in order to reduce the impact which preanaly-
tical sample handling may exert on RNA quality and sta-
bility. This is particularly important when collecting blood
samples for large-scale biobanks, where the associated
costs are very high. The overall goal of this study was to establish a prac-
tical protocol for sampling, handling, transportation and
storage of adult and cord blood RNA. On this back-
ground, we systematically compared the RNA quality,
quantity, and the RNA transcript stability of two com-
mercially available RNA stabilizing technologies PAX-
gene™Blood RNA system (PreAnalytiX, QIAGEN/BD,
Hombrechtikon, Switzerland), and Tempus™Blood RNA
system (Applied Biosystems, Foster City, CA). We then
investigated the effects of transportation and blood
volumes on the RNA transcript stability of six selected
genes (CDKN1A, FOS, IL8, MYC, IL1B, and TP53) using
quantitative real-time PCR (RT-qPCR). These genes
were selected based on literature search [7,8]. In this study, we investigated important input factors
for large-scale collection of blood from adults and chil-
dren (umbilical cord blood). Although high-quality RNA
can be prepared using fresh blood that is processed im-
mediately, this option is hardly realistic when sampling
for a large-scale biobank. Comparison of two RNA stabilizing systems, PAXgene and
Tempus Comparison of two RNA stabilizing systems, PAXgene and
Tempus Two commercially available blood RNA stabilizing sys-
tems, the PAXgene blood RNA system and Tempus
blood RNA system, were available as evacuated tubes
containing RNA stabilizing reagents, and both PAXgene
[7,11,12] and Tempus [13] blood RNA systems have
been shown to provide efficient stabilization of the
blood RNA for several days at room temperature. In this
study, the two systems were systematically evaluated
regarding their performance to stabilize blood RNA; i.e. preserving the RNA quality, yield, and the RNA tran-
script stability of four selected genes (CDKN1A, IL8,
MYC, and TP53). The study design for blood collection
is outlined in Figure 1. The RNA qualities and yields for
samples collected directly into PAXgene or Tempus Results and discussion In MoBa, the combination of biological specimens and
questionnaire data on lifestyle and exposures provide
unique possibilities to study the effects of many factors
of relevance for pregnancy outcomes and health [1]. Blood-based large biobanks such as MoBa, and multi-
center studies, increasingly incorporate gene expression
profiling studies in order to get insight into the bio-
logical
mechanisms
triggered
by
gene-environment
interactions and disease. However, collection, transporta-
tion, processing and storage of blood samples may repre-
sent logistical and technical challenges [9,10]. Therefore, Adult blood
(Venous blood from healthy
adult donors)
Cord blood
(Umbilical cord of newborns whose mothers
have given their informed consent to participate
in the Mother and Child (MoBa) Study)
1.RNA yield, purity and integrity by NanoDrop™ and by Agilent Bioanalyzer
2.RNA transcript stability by RT-qPCR
PAXgene™ tubes (n=20)
Tempus™ tubes (n=20)
Suboptimal blood volume QC:
Adult blood from one healthy donor was collected in Tempus™ tubes (n = 18)
Blood volume (ml) collected in Tempus™ tubes: 3.0, 2.5, 2.0, 1.5, 1.0 and 0.5
3 parallels for each volume. Transportation QC: Tempus (n=20) tubes
Adult blood from one healthy donor was collected in Tempus™ tubes (n = 20). Samples were divided randomly into five groups: 4 groups were sent by
standard mail to four different hospitals in Norway: Kirkenes, Bergen,
Kristiansand, Bærumand the fifth group was kept at Oslo (NIPH, lab). Temperature was monitored during transportation. Samples were frozen upon arrival at NIPH. RNA stabilizing technologies comparison:
Adult blood (n=40) tubes from one healthy donor
Stored 0, 2, 5, 7 days at RT (~22 °C)
Frozen at -20 °C overnight and then stored at -80
°C until extraction
RNA stabilizing technologies comparison:
Cord blood (n = 15) tubes from 8 different donors,
transported from one hospital to NIPH within 1 day. Samples were frozen upon arrival at NIPH at -20 °C
overnight and then stored at -80 °C until extraction
Suboptimal blood volume QC:
Cord blood from one donor was collected in Tempus™ tubes (n = 10). Blood volume (ml) collected in Tempus™ tubes: 3.3, 3.0, 2.0, 1.0, 0.5 ml. 2 parallels for each volume. PAXgene™ tubes (n=8)
Tempus™ tubes (n=7)
RNA quality assessment:
Figure 1 Flow chart: experimental design. Results and discussion 0
5
10
15
20
25
30
0
2
5
7
Cord blood
RNA (ug/ml blood)
Storage at RT (days)
A) Total RNA yield per ml blood
PAXgene
Tempus
Adult-blood
-1.5
-1.0
-0.5
0.0
0.5
1.0
1.5
7
5
2
0
Fold change
Storage at RT (days)
B) Adult blood samples collected in the PAXgene tubes
IL8
MYC
TP53
CDKN1A
-1.5
-1.0
-0.5
0.0
0.5
1.0
1.5
7
5
2
0
Fold change
Storage at RT (days)
C) Adult blood samples collected in the Tempus tubes
IL8
MYC
TP53
CDKN1A
0
0.5
1
1.5
2
CDKN1A
IL8
MYC
TP53
Normalized expression value
D) Cord blood samples collected in Tempus or PAXgene tubes
PAXgene (n=8)
Tempus (n=7) 0
5
10
15
20
25
30
0
2
5
7
Cord blood
RNA (ug/ml blood)
Storage at RT (days)
A) Total RNA yield per ml blood
PAXgene
Tempus
Adult-blood -1.5
-1.0
-0.5
0.0
0.5
1.0
1.5
7
5
2
0
Fold change
Storage at RT (days)
B) Adult blood samples collected in the PAXgene tubes
IL8
MYC
TP53
CDKN1A B) B) Adult blood samples collected in the PAXgene tubes tubes are shown in Figure 2A and Table 1. The average
total RNA yields for adult and cord blood samples col-
lected in the PAXgene tubes were 2.1 ± 0.2 μg and
7.8 ± 1.7 μg per ml blood, respectively, while the average
total RNA yields for samples collected in the Tempus tu-
bes were 5.3 ± 0.2 μg per ml blood for adult blood and
21.6± 3.8 μg per ml blood for cord blood (Figure 2A). The
difference in yield per ml blood between the two types of
collection tubes was significant (p < 0.05). The obtained
RNA yields for blood samples collected in the Tempus
tubes were consistently higher than for samples collected
in the PAXgene tubes (Figure 2A). Higher RNA yields from
the Tempus tubes compared with PAXgene tubes have
been previously reported [14,15]. The RNA yield obtained
from cord blood samples was 3–4 times higher than from
adult blood using both types of tubes (Figure 2A). Results and discussion Cord blood
(Umbilical cord of newborns whose mothers
have given their informed consent to participate
in the Mother and Child (MoBa) Study) (Umbilical cord of newborns whose mothers
have given their informed consent to participate
in the Mother and Child (MoBa) Study) RNA stabilizing technologies comparison:
Cord blood (n = 15) tubes from 8 different donors,
transported from one hospital to NIPH within 1 day. Samples were frozen upon arrival at NIPH at -20 °C
overnight and then stored at -80 °C until extraction Tempus™ tubes (n=7) Suboptimal blood volume QC:
Cord blood from one donor was collected in Tempus™ tubes (n = 10). Blood volume (ml) collected in Tempus™ tubes: 3.3, 3.0, 2.0, 1.0, 0.5 ml. 2 parallels for each volume. Page 3 of 9 Duale et al. BMC Research Notes 2012, 5:510
http://www.biomedcentral.com/1756-0500/5/510 Figure 2 Comparison of PAXgene and Tempus tubes. Blood
(adult or cord blood) was collected either in the PAXgene tubes or in
the Tempus tubes, and stored at room temperature for the indicated
number of days, followed by total RNA isolation. Five tubes were used
at each time point (n = 5). A) The RNA yield from the cord blood and
the adult blood samples collected either in the PAXgene tubes or in
the Tempus tubes. Each bar represents the average RNA yield per
tube ± SE. B) Relative transcript levels of four genes from adult blood
samples collected in the PAXgene tubes and stored for up to seven
days at room temperature; C) Relative transcript levels of four genes
from adult blood samples collected in the Tempus tubes and stored
for up to seven days at room temperature. The 0 day samples were
used as reference samples (calibrators); hence all other samples were
compared against the reference samples. Each bar represents the
average log2-transformed fold change values and the error bar
indicates ± SE. The dashed lines indicate ± 2 – fold change. The fold
change detected for the genes were within ± 2 – fold. D) The
normalized expression values of four genes from cord blood samples
collected in the PAXgene tubes (n = 8) and in the Tempus tubes
(n = 7). Each bar represents the normalized expression values
[2-ΔCq (sample); where the ΔCq (sample) ± Cq (gene) – Cq (internal
control)] and the error bar indicates ± SE. Results and discussion Storage at RT (days) -1.5
-1.0
-0.5
0.0
0.5
1.0
1.5
7
5
2
0
Fold change
Storage at RT (days)
C) Adult blood samples collected in the Tempus tubes
IL8
MYC
TP53
CDKN1A C) C) Adult blood samples collected in the Tempus tubes The integrity and purity of RNA can be used to evalu-
ate the quality of sample handling and processing. A
RIN-value higher than five is considered as indicating
good quality RNA whereas RIN-values above eight are
considered
ideal
for
downstream
applications
[4,5]. When we investigated the RNA quality, we observed
similar and comparable average OD 260/280 ratios and
average RIN values for all adult and cord blood samples
collected in Tempus and PAXgene tubes (p > 0.05), but
the average OD 260/230 ratio for the samples collected
in the PAXgene tubes was lower (p < 0.05) than for sam-
ples collected in Tempus tubes (Table 1). It has previ-
ously been reported that OD 260/230 ratios are higher
for Tempus tubes compared with PAXgene tubes [15]. The observed differences in the average OD 260/230
ratio between the two systems may be due to the
PAXgene elution buffer which may contain high salt
contents. Storage of PAXgene and Tempus samples at 0
0.5
1
1.5
2
CDKN1A
IL8
MYC
TP53
Normalized expression value
D) Cord blood samples collected in Tempus or PAXgene tubes
PAXgene (n=8)
Tempus (n=7) D) D) Cord blood samples collected in Tempus or PAXgene tubes Page 4 of 9 Page 4 of 9 Duale et al. Results and discussion BMC Research Notes 2012, 5:510
http://www.biomedcentral.com/1756-0500/5/510 Table 1 Comparison of blood samples collected in
PAXgene and in Tempus tubes
A) Adult blood
Storage at
RT (days)
RIN value
OD
260/280 ratio
OD
260/230 ratio
Number of
tubes
PAXgenes tube
0
8.0 ± 0.3
2.12 ± 0.02
1.56 ± 0.31
5
2
8.0 ± 0.3
2.10 ± 0.03
1.56 ± 0.62
5
5
8.2 ± 0.3
2.12 ± 0.02
1.37 ± 0.73
5
7
8.2 ± 0.3
2.09 ± 0.03
1.42 ± 0.70
5
Average
8.10 ± 0.3
2.11 ± 0.02
1.48 ± 0.61*
20
Tempus tubes
0
8.0 ± 0.3
2.12 ± 0.07
2.01 ± 0.14
5
2
8.2 ± 0.3
2.12 ± 0.07
2.01 ± 0.14
5
5
8.0 ± 0.3
2.14 ± 0.08
2.09 ± 0.08
5
7
8.2 ± 0.3
2.11 ± 0.04
2.08 ± 0.06
5
Average
8.10 ± 0.3
2.12 ± 0.07
2.05 ± 0.11
20
B) Cord blood
PAXgene
(n = 8)
Tempus
(n = 7)
RIN value
8.1 ± 0.2
8.0 ± 0.2
OD 260/280
ratio
2.1 ± 0.1
2.0 ± 0.3
OD 260/230
ratio
1.6 ± 0.5*
2.0 ± 0.8
Notes. * RNA from PAXgene tube-collected samples have significantly lower
OD 260/230 ratio compared to RNA from Tempus tube-collected samples,
p < 0.05. The cord blood samples were transported to NIPH (lab) within one
day. n = number of tubes. Table 1 Comparison of blood samples collected in
PAXgene and in Tempus tubes (Figure 2B and C). The non-normalized raw Cq-values for
the four genes also illustrate similar trends (Additional file
1A and B). The differences in the Cq-values between stored
and day 0 samples for each gene were very small. For cord
blood samples, the average transcript levels for the four
genes were comparable between the Tempus and the
PAXgene tube samples, and the difference between the two
systems (Figure 2D and Additional file 1C) was not signifi-
cant (p> 0.05). Storage for up to 7 days exceeds the
maximum period (3–5 days) as recommended by both
manufacturers. The recommended storage time may be
too strict, and a more flexible protocol – also giving RNA
of acceptable quality - can more conveniently be imple-
mented in multi-center studies. Results and discussion Given the results of this initial investigation where
samples collected in the Tempus tubes gave higher RNA
yields
with
comparable
quality
when
compared
to
PAXgene tubes, we selected the Tempus Blood RNA
system for MoBa and began collecting cord blood in
Tempus tubes in June 2005. Since then, approximately
50,000 samples have been collected and stored at −80°C. Transportation QC: The effect of transportation on RNA
quality and transcript stability The maternity units where sample collection took place
are located all over Norway, and from a few hours up to
several days may be required before the blood samples
arrive for processing at the NIPH biobank. The distance
between blood collection location and the biobank, and
the associated differences in duration and temperature
fluctuations during transportation, may affect the quality
of the obtained RNA and subsequently the gene expres-
sion data. We investigated the effect of transportation
on RNA quality and transcript stability of six genes
(CDKN1A, FOS, IL8, MYC, IL1B, and TP53) which have
been used in earlier studies [7,8]. Whole venous blood
from one healthy donor (Figure 1) was collected by ven-
ous puncture into 20 Tempus tubes. The tubes were
divided randomly into five groups, of which four were
sent by standard mail to four maternity units in Norway
whereas the fifth was kept at room temperature (24.4 ±
1.9°C) in the NIPH laboratory in Oslo. Samples were
returned from the maternity units to NIPH and kept
there at room temperature until all four sets had arrived;
they were then frozen at −20°C at the same time fol-
lowed by transfer to −80°C 24 hours later. The tem-
perature logs showed values during transportation ranging
from 3.4°C to 27.1°C; none of the samples were - or had
been - frozen (Additional file 2A). Samples sent to Kristian-
sand had the highest mean temperature (20.28± 5.58°C),
while those to Bærum had the lowest mean temperature
(16.96± 8.55°C). The mean temperature differences be-
tween the transported samples and samples kept at NIPH
are presented in Additional file 2A. Samples sent to Bærum Notes. * RNA from PAXgene tube-collected samples have significantly lower
OD 260/230 ratio compared to RNA from Tempus tube-collected samples,
p < 0.05. The cord blood samples were transported to NIPH (lab) within one
day. n = number of tubes. Notes. * RNA from PAXgene tube-collected samples have significantly lower
OD 260/230 ratio compared to RNA from Tempus tube-collected samples,
p < 0.05. The cord blood samples were transported to NIPH (lab) within one
day. n = number of tubes. room temperature for up to seven days had little effect on
RNA quality, although a slight decrease in RNA yield
(p < 0.05) was observed for adult samples collected in the
PAXgene tubes at seven days of storage (Figure 2A). Transportation QC: The effect of transportation on RNA
quality and transcript stability Compared to the
other samples, those from Bærum had much lower
temperature (≤6°C) for two 12-hour time intervals. This
might be relevant for the observed RNA quality of these had the highest mean temperature difference (−8.0°C) rela-
tive to the samples kept at NIPH. RNA was extracted from
all samples and analyzed for quality, and RT-qPCR was
carried out to measure the relative transcript levels for
the six selected genes. Figure 3A shows that trans-
portation of samples by standard mail apparently affected
RNA quality, especially the RNA integrity expressed as
RIN value. There was a statistically significant location-
dependent reduction in the average RIN value (p< 0.05);
samples kept at NIPH had the highest RIN values; the
lowest were from the Kirkenes and Bergen samples. The
temperature recordings were divided into 12-hour inter-
vals and averaged (Additional file 2A). Compared to the
other samples, those from Bærum had much lower
temperature (≤6°C) for two 12-hour time intervals. This
might be relevant for the observed RNA quality of these particular samples (Figure 3A and Additional file 2B). In
any case, the average RIN value for each group was greater
than six, which is considered acceptable for many RNA
assays. Figure 3B shows the relative transcript levels for the
six genes. Transportation of the samples apparently
affected the relative transcription levels of the genes. Samples sent to Kirkenes or Kristiansand showed the
highest changes compared to the samples kept at NIPH
(control samples) (Figure 3B). However, the observed
transcript level changes were again within ± 2–fold. The
non-normalized raw Cq-values for the six genes and the
average raw Cq-values ± SD for each gene are presented
in Additional file 2D. Differences in the Cq-values be-
tween transported samples and samples kept at NIPH
for each gene were small. A significant finding is that
higher RNA yields were obtained from transported sam-
ples compared to samples kept at NIPH (Figure 3A and
Additional file 2C); the reason for this is unclear. Sam-
ples can be shaken during transportation due to mech-
anical
movements
and
vibrations,
and
temperature
fluctuations occur (Additional file 2C), which may affect
the RNA extraction efficiency. Suboptimal blood volume QC: The effect of suboptimal
blood volume on RNA quality and transcript stability b ood
o u
e o
qua ty a d t a sc pt stab ty
Regular sample collections for the MoBa biobank were
carried out in busy maternity units. A considerable por-
tion of the samples collected contains blood volumes less
than the optimal volume recommended by the manufac-
turer (Figure 4A); this is a common problem when collec-
ting cord blood for research purposes. Figure 4A shows
cord blood collected in Tempus tubes from different ma-
ternity units during a period of one year (n= 10835). We
observed that ~ 20% of the tubes contained less than 2 ml
blood. We investigated whether the recommended blood–
to–RNA stabilizing reagent ratio was critical for blood
RNA quality and RNA transcript stability, and observed a
significant (p < 0.05) blood volume-dependent reduction
in RNA yield, RIN value, and OD 260/280 and OD 260/
230 ratios (Table 2A). The RNA blood chemistry seems to Figure 3 The effects of transportation on RNA, quantity, quality
d t
i t t bilit
Ad lt bl
d f
h
lth d Figure 3 The effects of transportation on RNA, quantity, quality
and transcript stability. Adult blood from one healthy donor was
ll
d
b
l
d
d d Figure 3 The effects of transportation on RNA, quantity, quality
and transcript stability. Adult blood from one healthy donor was
collected in Tempus™tubes (n = 20). Samples were divided
randomly into five groups (n = 4 tubes per group): four groups were
sent by standard mail to four different hospitals in Norway, and the
fifth group was kept at NIPH (Oslo/lab). Temperature was monitored
during transportation. A) The effects of transportation on RNA
quality and yield; B) The effects of transportation on RNA transcript
levels of six genes. Samples kept at NIPH were used as reference
samples; hence all other samples were compared against the
reference samples. Each bar represents the average
log2-transformed fold change values and the error bar
indicates ± SE. The dashed lines indicate ± 2 – fold change. g
y
y
and transcript stability. Adult blood from one healthy donor was
collected in Tempus™tubes (n = 20). Samples were divided
randomly into five groups (n = 4 tubes per group): four groups were
sent by standard mail to four different hospitals in Norway, and the
fifth group was kept at NIPH (Oslo/lab). Temperature was monitored
during transportation. Transportation QC: The effect of transportation on RNA
quality and transcript stability 0
2
4
6
8
10
12
14
16
RNA (µg/ 3 ml blood)
RIN
OD260/280
OD260/230
Quantity
Location
A) The effects of transportation on RNA quantity and quality
NIPH (Oslo)
Bærum
Kristiansand
Bergen
Kirkenes
-1.5
-1.0
-0.5
0.0
0.5
1.0
1.5
NIPH (Oslo)
Bærum
Kristiansand
Bergen
Kirkenes
Fold change
Location
B) The effects of transportation on relative transcript levels
CDKN1A
FOS
IL8
MYC
IL1B
TP53
Figure 3 The effects of transportation on RNA, quantity, quality
and transcript stability. Adult blood from one healthy donor was
collected in Tempus™tubes (n = 20). Samples were divided
randomly into five groups (n = 4 tubes per group): four groups were
sent by standard mail to four different hospitals in Norway, and the
fifth group was kept at NIPH (Oslo/lab). Temperature was monitored
during transportation. A) The effects of transportation on RNA
quality and yield; B) The effects of transportation on RNA transcript
levels of six genes. Samples kept at NIPH were used as reference
samples; hence all other samples were compared against the
reference samples. Each bar represents the average
log2-transformed fold change values and the error bar
indicates ± SE. The dashed lines indicate ± 2 – fold change. 0
2
4
6
8
10
12
14
16
RNA (µg/ 3 ml blood)
RIN
OD260/280
OD260/230
Quantity
Location
A) The effects of transportation on RNA quantity and quality
NIPH (Oslo)
Bærum
Kristiansand
Bergen
Kirkenes
B) A) Overall, transportation of the samples by standard
mail had some effects on RNA yield, quality and RNA
transcript stability. Temperature fluctuations, mechan-
ical movements or vibrations during the transportation
of samples may have contributed to the differences
(Figure 3 and Additional file 2). However, the observed
differences between transported samples and samples
kept at NIPH were minor, and the obtained RNA qual-
ity from transported samples was considered acceptable
for many RNA assays. Our results therefore suggest
that blood samples collected in Tempus tubes can be
sent by standard mail, under the conditions employed
in this study. B) B) The effects of transportation on relative transcript levels -1.5
-1.0
-0.5
0.0
0.5
1.0
1.5
NIPH (Oslo)
Bærum
Kristiansand
Bergen
Kirkenes
Fold change
Location
CDKN1A
FOS
IL8
MYC
IL1B
TP53 Transportation QC: The effect of transportation on RNA
quality and transcript stability The
RNAs isolated from both PAXgene and Tempus had aver-
age RIN-values of approximately eight, and average OD
260/A280 ratios were above 1.8 (Table 1). Based on these
data, high-quality RNA could be extracted from blood
samples stabilized with both PAXgene and the Tempus
system. room temperature for up to seven days had little effect on
RNA quality, although a slight decrease in RNA yield
(p < 0.05) was observed for adult samples collected in the
PAXgene tubes at seven days of storage (Figure 2A). The
RNAs isolated from both PAXgene and Tempus had aver-
age RIN-values of approximately eight, and average OD
260/A280 ratios were above 1.8 (Table 1). Based on these
data, high-quality RNA could be extracted from blood
samples stabilized with both PAXgene and the Tempus
system. The RNA transcript stability and potential alteration of
the transcript level for four genes (CDKN1A, IL8, MYC,
and TP53) were investigated by RT-qPCR (Figure 2B-D). Storage of both PAXgene and Tempus tubes at room
temperature for up to seven days had some effect on RNA
transcript stability; there were clear storage related changes
in the transcript levels. For adult blood samples collected
both in PAXgene and Tempus tubes, storage for up to
seven days at room temperature had effects on the relative
transcript levels for the four genes when compared to the
day 0 samples (Figure 2B and C). Storage effects were more
pronounced in samples collected in the PAXgene compared
to the Tempus tubes. However, the observed transcript
level changes were within ±2–fold for both types of tubes Page 5 of 9 Page 5 of 9 Duale et al. BMC Research Notes 2012, 5:510
http://www.biomedcentral.com/1756-0500/5/510 had the highest mean temperature difference (−8.0°C) rela-
tive to the samples kept at NIPH. RNA was extracted from
all samples and analyzed for quality, and RT-qPCR was
carried out to measure the relative transcript levels for
the six selected genes. Figure 3A shows that trans-
portation of samples by standard mail apparently affected
RNA quality, especially the RNA integrity expressed as
RIN value. There was a statistically significant location-
dependent reduction in the average RIN value (p< 0.05);
samples kept at NIPH had the highest RIN values; the
lowest were from the Kirkenes and Bergen samples. The
temperature recordings were divided into 12-hour inter-
vals and averaged (Additional file 2A). Suboptimal blood volume QC: The effect of suboptimal
blood volume on RNA quality and transcript stability A) The effects of transportation on RNA
quality and yield; B) The effects of transportation on RNA transcript
levels of six genes. Samples kept at NIPH were used as reference
samples; hence all other samples were compared against the
reference samples. Each bar represents the average
log2-transformed fold change values and the error bar
indicates ± SE. The dashed lines indicate ± 2 – fold change. Page 6 of 9 Duale et al. BMC Research Notes 2012, 5:510
http://www.biomedcentral.com/1756-0500/5/510 4495
2079
1676
996
642
862
85
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
3.0
2.5
2.0
1.5
1.0
0.5
0.0
Number of Tempus tubes
Blood volume (ml per tube)
A) Cord blood collected in Tempus tubes in MoBa for a period of one year
(n =1 0,835)
-10.0
-8.0
-6.0
-4.0
-2.0
0.0
2.0
3
2.5
2
1.5
1
0.5
Fold change
Blood volume (ml)
B) Suboptimal blood volume for Adult blood samples
CDKN1A
FOS
IL8
MYC
IL1B
TP53
-10.0
-8.0
-6.0
-4.0
-2.0
0.0
2.0
4.0
6.0
3.3
3
2
1
0.5
Fold Change
Blood volume (ml)
C) Suboptimal blood volume for cord blood samples
CDKN1A
FOS
IL8
MYC
IL1B
TP53 4495
2079
1676
996
642
862
85
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
3.0
2.5
2.0
1.5
1.0
0.5
0.0
Number of Tempus tubes
Blood volume (ml per tube)
A) Cord blood collected in Tempus tubes in MoBa for a period of one year
(n =1 0,835) Figure 4 The effect of suboptimal blood volume collected in
the Tempus tubes on RNA quality and transcript stability. A) Cord blood collected in Tempus tubes from maternity units in a
period of one year (n = 10 853). B) The effects of suboptimal volume
of adult blood collected in the Tempus tubes on relative transcript
level for six genes. Adult blood from one healthy donor was
collected in Tempus tubes (n = 18). The following blood volumes
(3.0, 2.5, 2.0, 1.5, 1.0 and 0.5 ml) were collected in the Tempus tubes
(n = 3; three parallels tubes for each blood volume). C) The effects of
suboptimal volume of cord blood collected in the Tempus tubes on
relative transcript level for six genes. Cord blood from one donor
was collected in Tempus tubes (n = 10). Suboptimal blood volume QC: The effect of suboptimal
blood volume on RNA quality and transcript stability The following blood
volumes (3.3, 3.0, 2.0, 1.0, and 0.5 ml) were collected in the Tempus
tubes (n = 2; two parallels tubes for each blood volume). Samples
with 3 ml blood per tube were used as reference samples; hence all
other samples were compared against the reference samples. Each
bar represents the average log2-transformed fold change values and
the error bar indicates = SE. Since some of the samples had low RNA
quality, selecting stably expressed reference genes for normalization
was a challenge. We therefore used the average Cq value for each
gene to normalize the samples: ΔCq (sample) = Cq (gene) – Cq
(average); these values where converted into linear scale, 2-ΔCq. Fold
changes were calculated by dividing ΔCq value of the sample by
the ΔCq value of the reference sample. -10.0
-8.0
-6.0
-4.0
-2.0
0.0
2.0
3
2.5
2
1.5
1
0.5
Fold change
Blood volume (ml)
B) Suboptimal blood volume for Adult blood samples
CDKN1A
FOS
IL8
MYC
IL1B
TP53 B) B) Suboptimal blood volume for Adult blood samples similar trend (Additional file 3A and B). Furthermore,
overfilling the Tempus tubes with cord-blood, i.e. more
than 3 ml blood per tube, seems to affect the apparent
RNA transcript stability, suggesting that the blood–to–
RNA
stabilizing
reagent
ratio
limit
was
exceeded
(Figure 4C). Our results indicate that the blood–to–RNA
stabilizing reagent ratio is very important for blood RNA
quality and transcript stability for adult blood samples,
whereas the blood volume is less critical for cord blood
samples. Hence, collection of 3 ml of blood per Tempus
tube will always be the highly recommended optimal
blood volume, but this volume is not always possible
when blood collection is carried out in a busy maternity
unit. For adult blood samples it is advisable to collect
3 ml blood per Tempus, whereas for cord blood samples,
minor deviations from the optimal volume per tube may
be acceptable. -10.0
-8.0
-6.0
-4.0
-2.0
0.0
2.0
4.0
6.0
3.3
3
2
1
0.5
Fold Change
Blood volume (ml)
C) Suboptimal blood volume for cord blood samples
CDKN1A
FOS
IL8
MYC
IL1B
TP53 C) C) Suboptimal blood volume for cord blood samples collapse when the blood volume in the tubes was below
2 ml. This was true for adult blood, but suboptimal blood
volumes were not critical when collecting cord blood
(Table 2B). Conclusions
l Our results indicate that blood samples in a large bio-
bank such as MoBa can be used to obtain high-quality
RNA suitable for gene expression analysis, provided that
special collection tubes are used. The quality of the RNA
isolated from such tubes is crucial for successful tran-
script profiling analyses. Transportation of samples by
standard mail apparently affected the RNA quality and
RNA transcript stability, but the changes were moderate
and the overall RNA quality of the transported samples
was good. Some unexplained changes in gene expression
changes seem to be associated with suboptimal blood
volumes collected in the tubes. Overall, we demonstrate
that satisfactory amounts of high-quality RNA can be The relative transcript levels for the six genes are
shown in Figure 4B and C. We observed blood–volume
dependent reduction in the relative transcript levels for
the six genes compared with samples with 3 ml blood
volume (Figure 4B and C). Samples with volumes less
than 1.5 ml for adult blood and less than 1.0 ml for cord
blood showed the highest changes in the relative tran-
script levels for the six genes, compared with the optimal
3 ml samples (Figure 4B and C). The non-normalized
raw Cq-values for adult and cord blood samples show a Duale et al. Conclusions
l BMC Research Notes 2012, 5:510
http://www.biomedcentral.com/1756-0500/5/510 Page 7 of 9 Table 2 The effect of blood volume in the Tempus tubes on RNA quality
Blood volume (ml)
Yield total RNA (ug)
RNA Integrity Number (RIN)
OD260/280 ratio
OD2602/230 ratio
A) Adult blood samples (n = 3)*
3.0
7.90 ± 1.50
8.20 ± 0.06
2.19 ± 0.02
2.10 ± 0.23
2.5
3.38 ± 0.90
8.13 ± 0.27
2.24 ± 0.05
0.91 ± 0.46
2.0
2.94 ± 0.91
8.40 ± 0.08
2.12 ± 0.08
1.05 ± 0.53
1.5
1.38 ± 0.59
3.53 ± 2.53
2.51 ± 0.08
0.64 ± 0.32
1.0
0.55 ± 0.50
1.13 ± 0.13
1.19 ± 0.44
0.38 ± 0.38
0.5
0.07 ± 0.03
1.00 ± 0.00
0.77 ± 0.39
0.05 ± 0.05
B) Cord blood samples (n = 2)
3.3
79.65 ± 4.76
7.15 ± 0.78
2.15 ± 0.00
2.12 ± 0.00
3.0
74.60 ± 1.23
7.50 ± 0.65
2.17 ± 0.00
2.15 ± 0.01
2.0
44.42 ± 1.91
9.10 ± 0.00
2.18 ± 0.01
2.09 ± 0.01
1.0
33.92 ± 3.13
9.45 ± 0.04
1.98 ± 0.01
1.61 ± 0.02
0.5
4.29 ± 0.11
1.03 ± 0.03
1.25 ± 0.03
0.53 ± 0.01
Notes. *There are significant blood volume dependent reductions in the RNA yields and in the RNA qualities (RIN, OD 260/280 and OD 260/230 ratio), p < 0.05. n = number of tubes. Table 2 The effect of blood volume in the Tempus tubes on RNA quality
Blood volume (ml)
Yield total RNA (ug)
RNA Integrity Number (RIN) achieved from samples routinely collected in a large bio-
bank. This is good news for future projects studying asso-
ciation of disease with altered patterns of gene expression. temperature at NIPH (Oslo/lab). The tubes were sent to
the hospitals and then returned by the local staff to
NIPH, to comprise a worst-case scenario of double the
ordinary mail transportation time. Upon arrival at NIPH
samples were kept at RT until the last sample arrived and
then they were all frozen at the same time. Temperature
was monitored during the transportation using small
sensors with memory (Tinytag Talk2, Precision Technic
Nordic, Rødovre, Denmark). The Regional Ethics Com-
mittee and the Data Inspectorate approved the MoBa
study, and participants gave their informed consent. RNA extraction
T
l RNA f Total RNA from whole blood collected in PAXgene
tubes was extracted according to the manufacturer’s ins-
tructions and included DNase I treatment (PreAnalytiX,
QIAGEN/BD, Hombrechtikon, Switzerland). Total RNA
from blood collected in Tempus tubes was extracted using
the Tempus™6-Port RNA Isolation Kit Protocol on an
ABI PRISMW TM 6100 Nucleic Acid PrepStation accor-
ding to the manufacturer’s instructions and included DNase
I treatment and addition of 1X PBS bringing the total vo-
lume to 12 ml prior to processing (Applied Biosystems,
Foster City, CA). An aliquot of 5 μl of each extracted total RNA was
used for RNA quality control assessments, while the
remaining RNA sample was stored at – 80°C until use. The adult blood samples collected to study the effects
of transportation were shipped to 4 different locations
in Norway, with instructions to treat the tubes the same
way (temperature, storage) as tubes collected in the
local maternity unit, and then to return the tubes to
NIPH as it would be normally done with the collected
tubes. The four sets were shipped by standard mail to
the maternity units in Bærum (~15 km from Oslo),
Kristiansand (~ 320 km), Bergen (~ 520 km) and Kir-
kenes (~ 2000 km). A fifth set of samples was let at room Blood collection and study design Whole blood was collected at a maternity unit in Oslo
from the umbilical cord of newborns whose mothers
had given their informed consent to participate in the
MoBa project. Venous blood samples were obtained via
phlebotomy from healthy adult donors among the NIPH
staff after informed consent. The study design for blood
collection is outlined in Figure 1. Samples were collected
either into Tempus™Blood RNA Tubes (space for 3 ml
blood) (Applied Biosystems, Foster City, CA) or PAX-
gene Blood RNA Tubes (space for 2.5 ml blood) (PreA-
nalytiX, Qiagen, Germany). Samples collected at NIPH
(Oslo) were stored for 2 h at room temperature, then at
– 20°C for 24 h and finally transferred to – 80°C for
long-term storage. Samples shipped by standard mail
were then stored at – 20°C for 24 h after arrival at
NIPH, and then transferred to – 80°C for long term
storage. All samples were handled as recommended by
the manufacturers prior to processing. Statistical analysis
l
l Statistical analysis
Statistical analysis of RNA yield, purity, integrity and
ΔCq - values was carried out by one-way analysis of
variance (ANOVA), followed by post hoc Dunnett’s tests
to allow for multiple comparisons or by non-parametric
Kruskal-Wallis test. Normal distribution and equality of
variances was evaluated using the Shapiro-Wilk test and
the Levene's test of homogeneity of variance. Two-
sample t-test was used to compare cord blood samples
collected in the PAXgene vs the Tempus tubes. The data
are presented as means ± SE. Statistical analyses were
performed using SPSS v17 software (SPSS, Inc., Chicago,
IL), and p < 0.05 was accepted as statistically significant. Quantitative real-time PCR (qPCR) was performed in
96-well PCR plates using a 7500Fast Real-Time PCR Sys-
tem (Applied Biosystems, Foster City, CA). The cDNA
samples were diluted 10-fold and gene-specific transcrip-
tion levels were determined in a 20 μl reaction volume in
triplicates using TaqManWFast PCR Universal PCR Mas-
ter mix following the manufacturer’s protocol. Commer-
cially available primers and probe assays were from
Applied Biosystems: IL1B (PN: Hs00174097_m1), IL8
(PN: Hs00174103_m1), FOS (PN: Hs00170630_m1), MYC
(PN: Hs00153408_m1), TP53 (PN: Hs00153340_m1), and
CDKN1A (PN: Hs00355782_m1), and 18S rRNA (PN:
Hs99999901_s1). The PCR cycling program included an
enzyme activation step at 95°C for 20 seconds, and then
40 cycles of annealing and extension steps at 95°C for
3 seconds and 60°C for 30 seconds, respectively. Additional files Additional file 1: The non-normalized raw Cq-values. A) The
non-normalized raw Cq-values for adult blood samples collected in the
PAXgene tubes; B) The non-normalized raw Cq-values for adult blood
samples collected in the Tempus tubes; C) The non-normalized raw
Cq-values for cord blood samples collected in the PAXgene and in the
Tempus tubes. Each bar represents the average Cq-values and the error
bar indicates ± SE. Additional file 2: Temperature recordings for transported samples
vs RIN, RNA yield and Raw Cq-value. Temperatures were monitored
during the transportation of the samples to four different locations
(hospitals) in Norway. The four samples were sent by standard mail at
February 21th and all samples returned to NIPH at February 24th. Transportation time for samples sent to Bærum, Kristiansand and Bergen
was ~ 48 hours, while it took ~ 74 hours for Kirkenes samples to return to
NIPH. The temperature variations ranged from 3.4°C to 27.1°C, and none
of the samples were frozen during the transportation. A) Temperature
recordings for samples sent to the four locations and samples kept at
NIPH; B) RIN values vs. average temperature (°C) of the samples; C) RNA
yields vs. average temperature (°C) of the samples; D) The non-
normalized raw Cq-values vs. average temperature (°C) of the samples. Each bar represents the average Cq-values and the error bar
indicates ± SE for the raw Cq-values or ± SD for average temperature (°C)
of the samples. RNA QC
h The concentration of extracted total RNA was measured
using NanoDrop ND-1000 spectrophotometer (Fisher
Scientific, Norway). RNA purity was estimated by exam-
ining the OD 260/280 and the OD 260/230 ratios. The
RNA integrity was assessed by Agilent 2100 Bioanalyzer Page 8 of 9 Duale et al. BMC Research Notes 2012, 5:510
http://www.biomedcentral.com/1756-0500/5/510 (calibrator); fold change = 2-ΔΔCq]. A reference sample was
determined within each study, and for samples without an
obvious reference sample (calibrator), the linear scale of
ΔCq (sample); 2-ΔCq was used [17,18]. The fold change
values were then log2-transformed in order to make the
values symmetrical around zero. The 18S rRNA stability
was evaluated and results are presented in Additional file 4. These four genes: CDKN1A, IL8, MYC, and TP53 were
analyzed when the two RNA stabilization systems (PAX-
gene and Tempus) were compared, and then two more
genes (FOS and ILB) were added, i.e. six genes were ana-
lyzed in the remaining study. using the Eukaryote total RNA 6000 Nano LabChip kit
and Eukaryote total RNA Nano assay according to the
manufacturer’s instructions (Agilent Technologies, Palo
Alto, CA). The RNA integrity numbers (RIN) were cal-
culated using the Agilent 2100 Expert Software (RIN = 1;
low RNA quality to RIN = 10; highest RNA quality). using the Eukaryote total RNA 6000 Nano LabChip kit
and Eukaryote total RNA Nano assay according to the
manufacturer’s instructions (Agilent Technologies, Palo
Alto, CA). The RNA integrity numbers (RIN) were cal-
culated using the Agilent 2100 Expert Software (RIN = 1;
low RNA quality to RIN = 10; highest RNA quality). Quantitative real-time PCR measurement
Total RNA (500 ng) from samples was used as template
for the synthesis of cDNA using the High Capacity
cDNA Reverse Transcription Kit (Applied Biosystems,
Foster City, CA) according to manufacturer’s protocol. The
amplification reactions were carried out in an Eppendorf
MasterCycler (Eppendorf, Hamburg, Germany) with the
following steps: 10 minutes at 25°C, 2 hours at 37°C and fi-
nally, 5 minutes at 85°C. The quality of the cDNA was
assessed by the NanoDrop ND-1000 spectrophotometer
(Fisher Scientific), and the cDNA was stored at - 20°C until
use. Additional file 4: The evaluation of 18S rRNA stability.
A) Comparison of PAXgene tube-collected samples against the Tempus p
g
The authors declare that they have no competing interests. 10. Holland NT, Smith MT, Eskenazi B, Bastaki M: Biological sample collection
and processing for molecular epidemiological studies. Mutat Res 2003,
543:217–234. The authors declare that they have no competing interests. Authors' contributions 11. Muller MC, Merx K, Weisser A, Kreil S, Lahaye T, Hehlmann R, et al:
Improvement of molecular monitoring of residual disease in leukemias
by bedside RNA stabilization. Leukemia 2002, 16:2395–2399. ND participated in the study design and experimental work, participated in
scientific discussions, carried out the data and the statistical analyses,
interpreted the results, drafted the manuscript and prepared the finale
version of the manuscript. GB participated in the study design, in scientific
discussions, interpretation of the results, manuscript preparation and
supervised the work. KR participated in the study design, in scientific
discussions and in the experimental work. TB participated in the study
design, in scientific discussions, and manuscript preparation. JA participated
in the experimental work. KA participated in the experimental work. PM
participated in scientific discussions and manuscript preparation. CS participated in scientific discussions and manuscript preparation. ES participated in scientific discussions and WL participated in the study
design, in scientific discussions and manuscript preparation and
co-supervised the work. All authors have read and approved the final version
of the manuscript. 12. Weber DG, Casjens S, Rozynek P, Lehnert M, Zilch-Schoneweis S, Bryk O,
et al: Assessment of mRNA and microRNA Stabilization in Peripheral
Human Blood for Multicenter Studies and Biobanks. Biomark Insights
2010, 5:95–102. 13. Prezeau N, Silvy M, Gabert J, Picard C: Assessment of a new RNA
stabilizing reagent (Tempus Blood RNA) for minimal residual disease in
onco-hematology using the EAC protocol. Leuk Res 2006, 30:569–574. 14. Matheson LA, Duong TT, Rosenberg AM, Yeung RS: Assessment of sample
collection and storage methods for multicenter immunologic research in
children. J Immunol Methods 2008, 339:82–89. 15. Asare AL, Kolchinsky SA, Gao Z, Wang R, Raddassi K, Bourcier K, et al:
Differential gene expression profiles are dependent upon method of
peripheral blood collection and RNA isolation. BMC Genomics 2008, 9:474. g
p p p
co-supervised the work. All authors have read and approved the final version
of the manuscript. 16. Lefever S, Hellemans J, Pattyn F, Przybylski DR, Taylor C, Geurts R, et al:
RDML: structured language and reporting guidelines for real-time
quantitative PCR data. Nucleic Acids Res 2009, 37:2065–2069. Acknowledgements This study was supported by ABC (US National Institutes of Health award
NS047537) and NewGeneris (Contract No 016320–2). MoBa is supported by
the Norwegian Ministry of Health, the Ministry of Education and Research,
NIH/NIEHS (contract no. NO1-ES-85433), NIH/NINDS (grant no.1 UO1 NS
047537–01), and the Norwegian Research Council /FUGE (grant no. 151918/
S10). We thank Jorid Eide for help with collection of the cord blood
specimens. 17. Livak KJ, Schmittgen TD: Analysis of relative gene expression data using
real-time quantitative PCR and the 2(T)(−Delta Delta C) method. Methods
2001, 25:402–408. 18. Schmittgen TD, Livak KJ: Analyzing real-time PCR data by the
comparative C(T) method. Nat Protoc 2008, 3:1101–1108. doi:10.1186/1756-0500-5-510
Cite this article as: Duale et al.: Human blood RNA stabilization in
samples collected and transported for a large biobank. BMC Research
Notes 2012 5:510. doi:10.1186/1756-0500-5-510
Cite this article as: Duale et al.: Human blood RNA stabilization in
samples collected and transported for a large biobank. BMC Research
Notes 2012 5:510. We also thank Applied Biosystems in Norway and Sweden (Tempus™) and
VWR Norway (PAXgene™) for supplying blood RNA stabilizing collection
tubes and associated kits during the comparison of the two blood RNA
stabilizing systems study. VWR Norway was PAXgene™Blood RNA system
distributor in Norway at that time. Data analysis
h
f tube-collected samples: The Cq – values for 18S rRNA were relatively
stable following the storage of the tubes for up to seven days at room
temperature for both tubes; B) Evaluation of 18S rRNA stability for
transportation QC samples: The Cq – values for 18S rRNA were relatively
stable following the transportation; C) Suboptimal blood volume in the
Tempus tubes on 18S rRNA expression stability for adult blood samples:
Suboptimal blood volume collected in the tube has a significant effects
on 18S rRNA expression stability; and D) Suboptimal blood volume in the
Tempus tubes on 18S rRNA expression stability for cord blood samples:
Suboptimal blood volume collected in the tube has some effects on 18S
rRNA expression stability. Each bar represents the average Cq-values and
the error bar indicates ± SE. 4. Fleige S, Walf V, Huch S, Prgomet C, Sehm J, Pfaffl MW: Comparison of
relative mRNA quantification models and the impact of RNA integrity in
quantitative real-time RT-PCR. Biotechnol Lett 2006, 28:1601–1613. 5. Fleige S, Pfaffl MW: RNA integrity and the effect on the real-time qRT-PCR
performance. Mol Aspects Med 2006, 27:126–139. 6. Thach DC, Lin BC, Walter E, Kruzelock R, Rowley RK, Tibbetts C, et al:
Assessment of two methods for handling blood in collection tubes with
RNA stabilizing agent for surveillance of gene expression profiles with
high density microarrays. J Immunol Methods 2003, 283:269–279. 7. Rainen L, Oelmueller U, Jurgensen S, Wyrich R, Ballas C, Schram J, et al:
Stabilization of mRNA expression in whole blood samples. Clin Chem
2002, 48:1883–1890. Received: 27 April 2012 Accepted: 13 September 2012
Published: 18 September 2012 • Thorough peer review Abbreviations
f 8. 8. Baechler EC, Batliwalla FM, Karypis G, Gaffney PM, Moser K, Ortmann WA,
et al: Expression levels for many genes in human peripheral blood cells
are highly sensitive to ex vivo incubation. Genes Immun 2004, 5:347–353. Cq: Quantification cycle Threshold; RT-qPCR: Quantitative Real-time PCR;
RIN: RNA Integrity Number; MoBa: The Norwegian Mother and Child Cohort
Study; NIPH: Norwegian Institute of Public Health; QC: Quality Control. 9. Holland NT, Pfleger L, Berger E, Ho A, Bastaki M: Molecular epidemiology
biomarkers–sample collection and processing considerations. Toxicol Appl
Pharmacol 2005, 206:261–268. Author details
1Th N
i 1The Norwegian Institute of Public Health, Oslo, Norway. 2The Mailman
School of Public Health, Columbia University, New York, NY, USA. 3New York
State Psychiatric Institute, New York, NY, USA. 4Present address: Oslo
University Hospital, Rikshospitalet, Oslo, Norway. Submit your next manuscript to BioMed Central
and take full advantage of:
• Convenient online submission
• Thorough peer review
• No space constraints or color figure charges
• Immediate publication on acceptance
• Inclusion in PubMed, CAS, Scopus and Google Scholar
• Research which is freely available for redistribution
Submit your manuscript at
www.biomedcentral.com/submit Submit your next manuscript to BioMed Central
and take full advantage of:
• Convenient online submission
• Thorough peer review
• No space constraints or color figure charges
• Immediate publication on acceptance
• Inclusion in PubMed, CAS, Scopus and Google Scholar
• Research which is freely available for redistribution
Submit your manuscript at
www.biomedcentral.com/submit Submit your next manuscript to BioMed Central
and take full advantage of: Data analysis
h
f The quantification cycle (Cq; standard name for Ct and
Cp [16]) values were recorded with SDS v1.3 software
(Applied Biosystems, Foster City, CA); the Cq value is
the fractional cycle number at which the fluorescence
exceeds a fixed threshold. The raw Cq-values were then
exported into Excel- files and analyzed by the compara-
tive Cq – method [17,18] using 18S rRNA as internal
control. Targets with Cq-values > 35 were considered be-
yond the limit of detection (LOD) and all Cq-values
above 35 were coded as missing values. For downstream
analysis missing values were replaced by Cq (LOD) + 1
(i.e., 35 + 1: Cq = 36). Target genes were normalized to
18S rRNA internal controls, [this is given by ΔCq; where
ΔCq (sample) = Cq (target gene) – Cq (internal controls)]. The ΔΔCq values were generated by subtracting the ΔCq-
value for the reference samples (calibrators) from the
ΔCq-value for the samples [ΔΔCq = ΔCq (sample) – ΔCq Additional file 3: The non-normalized raw Cq-values for suboptimal
blood volume QC. A) Evaluation of the non-normalized raw Cq-values
for suboptimal blood volume QC from adult blood samples; and B)
Evaluation of the non-normalized raw Cq-values for suboptimal blood
volume QC from cord blood samples. Each bar represents the average
Cq-values and the error bar indicates ± SE. Additional file 4: The evaluation of 18S rRNA stability. A) Comparison of PAXgene tube-collected samples against the Tempus Additional file 4: The evaluation of 18S rRNA stability. A) Comparison of PAXgene tube-collected samples against the Tempus Page 9 of 9 Page 9 of 9 Duale et al. BMC Research Notes 2012, 5:510
http://www.biomedcentral.com/1756-0500/5/510 Duale et al. BMC Research Notes 2012, 5:510
http://www.biomedcentral.com/1756-0500/5/510 3. Hartel C, Bein G, Muller-Steinhardt M, Kluter H: Ex vivo induction of
cytokine mRNA expression in human blood samples. J Immunol Methods
2001, 249:63–71. Submit your next manuscript to BioMed Central
and take full advantage of: Submit your next manuscript to BioMed Central
and take full advantage of:
• Convenient online submission
• Thorough peer review
• No space constraints or color figure charges
• Immediate publication on acceptance
• Inclusion in PubMed, CAS, Scopus and Google Scholar
• Research which is freely available for redistribution
Submit your manuscript at
www.biomedcentral.com/submit • Convenient online submission Received: 27 April 2012 Accepted: 13 September 2012
Published: 18 September 2012 References 1. Magnus P, Irgens LM, Haug K, Nystad W, Skjaerven R, Stoltenberg C: Cohort
profile: the Norwegian Mother and Child Cohort Study (MoBa). Int J
Epidemiol 2006, 35:1146–1150. 1. Magnus P, Irgens LM, Haug K, Nystad W, Skjaerven R, Stoltenberg C: Cohort
profile: the Norwegian Mother and Child Cohort Study (MoBa). Int J
Epidemiol 2006, 35:1146–1150. p
2. Ronningen KS, Paltiel L, Meltzer HM, Nordhagen R, Lie KK, Hovengen R, et
al: The biobank of the Norwegian Mother and Child Cohort Study: a
resource for the next 100 years. Eur J Epidemiol 2006, 21:619–625.
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Scaffold Geometry-Imposed Anisotropic Mechanical Loading Guides the Evolution of the Mechanical State of Engineered Cardiovascular Tissues in vitro
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Document status and date:
Published: 16/02/2022 Document status and date:
Published: 16/02/2022 Document Version:
Publisher’s PDF, also known as Version of Record (includes final page, issue and volume numbers) Document Version:
Publisher’s PDF, also known as Version of Record (includes final page, issue and volume numbers) Scaffold Geometry-Imposed Anisotropic Mechanical Loading
Guides the Evolution of the Mechanical State of Engineered
Cardiovascular Tissues in vitro.
Citation for published version (APA):
Hermans, L. H. L., van Kelle, M. A. J., Oomen, P. J. A., Lopata, R. G. P., Loerakker, S., & Bouten, C. V. C.
(2022). Scaffold Geometry-Imposed Anisotropic Mechanical Loading Guides the Evolution of the Mechanical
State of Engineered Cardiovascular Tissues in vitro. Frontiers in Bioengineering and Biotechnology, 10, 796452.
Article 796452. https://doi.org/10.3389/fbioe.2022.796452 Scaffold Geometry-Imposed Anisotropic Mechanical Loading
Guides the Evolution of the Mechanical State of Engineered
Cardiovascular Tissues in vitro. Citation for published version (APA):
Hermans, L. H. L., van Kelle, M. A. J., Oomen, P. J. A., Lopata, R. G. P., Loerakker, S., & Bouten, C. V. C. (2022). Scaffold Geometry-Imposed Anisotropic Mechanical Loading Guides the Evolution of the Mechanical
State of Engineered Cardiovascular Tissues in vitro. Frontiers in Bioengineering and Biotechnology, 10, 796452. Article 796452. https://doi.org/10.3389/fbioe.2022.796452 Scaffold Geometry-Imposed Anisotropic Mechanical Loading
Guides the Evolution of the Mechanical State of Engineered
Cardiovascular Tissues in vitro. Citation for published version (APA):
Hermans, L. H. L., van Kelle, M. A. J., Oomen, P. J. A., Lopata, R. G. P., Loerakker, S., & Bouten, C. V. C. (2022). Scaffold Geometry-Imposed Anisotropic Mechanical Loading Guides the Evolution of the Mechanical
State of Engineered Cardiovascular Tissues in vitro. Frontiers in Bioengineering and Biotechnology, 10, 796452. Article 796452. https://doi.org/10.3389/fbioe.2022.796452 Citation for published version (APA):
Hermans, L. H. L., van Kelle, M. A. J., Oomen, P. J. A., Lopata, R. G. P., Loerakker, S., & Bouten, C. V. C. (2022). Scaffold Geometry-Imposed Anisotropic Mechanical Loading Guides the Evolution of the Mechanical
State of Engineered Cardiovascular Tissues in vitro. Frontiers in Bioengineering and Biotechnology, 10, 796452. Article 796452. https://doi.org/10.3389/fbioe.2022.796452 DOI:
10.3389/fbioe.2022.796452 DOI:
10.3389/fbioe.2022.796452 Document status and date:
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follow below link for the End User Agreement: Scaffold Geometry-Imposed
Anisotropic Mechanical Loading
Guides the Evolution of the
Mechanical State of Engineered
Cardiovascular Tissues in vitro
L. H. L. Hermans 1,2, M. A. J. Van Kelle 1,2, P. J. A. Oomen 1,2, R .G. P. Lopata 1, S. Loerakker 1,2*†
and C. V. C. Bouten 1,2†
1Department of Biomedical Engineering Eindhoven University of Technology Eindhoven Netherlands
2Institute for Complex L. H. L. Hermans 1,2, M. A. J. Van Kelle 1,2, P. J. A. Oomen 1,2, R .G. P. Lopata 1, S. Loerakker 1,2*†
and C. V. C. Bouten 1,2† 1Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, Netherlands, 2Institute for Complex
Molecular Systems, Eindhoven University of Technology, Eindhoven, Netherlands www.tue.nl/taverne Take down policy
If you believe that this document breaches copyright please contact us at:
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l providing details and we will investigate your claim. Download date: 24. Oct. 2024 ORIGINAL RESEARCH
published: 16 February 2022
doi: 10.3389/fbioe.2022.796452 Keywords: cardiovascular, tissue engineering, growth, remodeling, scaffold geometry Reviewed by: Reviewed by:
Joao Silva Soares,
Virginia Commonwealth University,
United States
Agneta Simionescu,
Clemson University, United States
*Correspondence:
S. Loerakker
s.loerakker@tue.nl
†These authors share last authorship Reviewed by:
Joao Silva Soares,
Virginia Commonwealth University,
United States
Agneta Simionescu,
Clemson University, United States Specialty section:
This article was submitted to
Biomechanics,
a section of the journal
Frontiers in Bioengineering and
Biotechnology
Received: 16 October 2021
Accepted: 28 January 2022
Published: 16 February 2022 Specialty section:
This article was submitted to
Biomechanics,
a section of the journal
Frontiers in Bioengineering and
Biotechnology Specialty section:
This article was submitted to
Biomechanics,
a section of the journal
Frontiers in Bioengineering and
Biotechnology
Received: 16 October 2021
Accepted: 28 January 2022
Published: 16 February 2022
Citation:
Hermans LHL, Van Kelle MAJ,
Oomen PJA, Lopata RGP, Loerakker S
and Bouten CVC (2022) Scaffold Cardiovascular tissue engineering is a promising approach to develop grafts that, in contrast to
current replacement grafts, have the capacity to grow and remodel like native tissues. This
approach largely depends on cell-driven tissue growth and remodeling, which are highly
complex processes that are difficult to control inside the scaffolds used for tissue engineering. For several tissue engineering approaches, adverse tissue growth and remodeling outcomes
were reported, such as aneurysm formation in vascular grafts, and leaflet retraction in heart
valve grafts. It is increasingly recognized that the outcome of tissue growth and remodeling,
either physiological or pathological, depends at least partly on the establishment of a
homeostatic mechanical state, where one or more mechanical quantities in a tissue are
maintained in equilibrium. To design long-term functioning tissue engineering strategies,
understanding how scaffold parameters such as geometry affect the mechanical state of a
construct, and how this state guides tissue growth and remodeling, is therefore crucial. Here,
we studied how anisotropic versus isotropic mechanical loading—as imposed by initial
scaffold geometry—influences tissue growth, remodeling, and the evolution of the
mechanical state and geometry of tissue-engineered cardiovascular constructs in vitro. Using a custom-built bioreactor platform and nondestructive mechanical testing, we
monitored the mechanical and geometric changes of elliptical and circular, vascular cell-
seeded, polycaprolactone-bisurea scaffolds during 14 days of dynamic loading. The elliptical
and circular scaffold geometries were designed using finite element analysis, to induce
anisotropic and isotropic dynamic loading, respectively, with similar maximum stretch
when cultured in the bioreactor platform. We found that the initial scaffold geometry-
induced (an)isotropic loading of the engineered constructs differentially dictated the
evolution of their mechanical state and geometry over time, as well as their final structural
organization. These findings demonstrate that controlling the initial mechanical state of tissue-
engineered constructs via scaffold geometry can be used to influence tissue growth and
remodeling and determine tissue outcomes. Edited by:
Andrea Malandrino,
Universitat Politecnica de Catalunya,
Spain 1 INTRODUCTION monitor the mechanical and geometric state of these constructs
over time. In previous studies, this platform was limited to applying
isotropic deformation, whereas most cardiovascular tissues,
including heart valves and blood vessels, are deformed in an
anisotropic manner (Zhou and Fung, 1997; Billiar and Sacks,
2000). We therefore implemented anisotropic deformation in
the Vertigro bioreactor by introducing an elliptical scaffold
geometry that was designed using finite element analysis. With
the same approach, circular control scaffolds were designed to
undergo isotropic deformation of similar magnitude. In this way,
we aimed to characterize how initial scaffold geometry, resulting in
either anisotropic or isotropic deformation, affects tissue growth
and remodeling, and the evolution of the mechanical state and
geometry of tissue-engineered constructs over time. Elliptical and
circular tissue-engineered constructs were dynamically loaded at
1 Hz with a peak pressure of 6 kPa for 14 days in the Vertigro
bioreactor. The mechanical state and geometry of the constructs
were characterized on day 0, 4, 7, 11, and 14 of dynamic loading
using ultrasound imaging and inverse finite element analysis. The
mechanical state was characterized in terms of stress, strain, strain
energy density, and stiffness in the central 50% of each construct. After 14 days of dynamic loading, the final tissue structure and
composition were assessed. We found that the initial scaffold
geometry-induced mechanical state of the tissue-engineered
constructs dictated the evolution of their mechanical state and
geometry over time, as well as their final structural organization. These results demonstrate that the initial mechanical state of
tissue-engineered
constructs
plays
an
important
role
in
determining the outcome of tissue growth and remodeling in vitro. When
conventional
treatments
fail,
end-stage
diseased
cardiovascular tissues like heart valves or blood vessels require
replacement. If autografts are unavailable, non-living prostheses
are commonly used. Such prostheses, like bioprosthetic and
mechanical valves, or synthetic vascular grafts, have several
shortcomings, but most importantly lack the ability to grow,
repair and remodel in response to changes in their environment. This aspect is particularly problematic for pediatric patients, as it
requires them to undergo multiple re-operations (Ackermann
et al., 2007; Lee et al., 2011; Brown et al., 2012) that reduce life
expectancy (Puvimanasinghe et al., 2001). In situ tissue engineering is a promising approach to develop a
new
generation
of
cardiovascular
grafts
that
potentially
overcomes the aforementioned drawback. Citation: Citation:
Hermans LHL, Van Kelle MAJ,
Oomen PJA, Lopata RGP, Loerakker S
and Bouten CVC (2022) Scaffold
Geometry-Imposed Anisotropic
Mechanical Loading Guides the
Evolution of the Mechanical State of
Engineered Cardiovascular
Tissues in vitro. Front. Bioeng. Biotechnol. 10:796452. doi: 10.3389/fbioe.2022.796452 February 2022 | Volume 10 | Article 796452 1 Frontiers in Bioengineering and Biotechnology | www.frontiersin.org Mechanical Evolution of Tissue-Engineered Constructs Hermans et al. 1 INTRODUCTION With this approach,
a (synthetic) biodegradable scaffold is implanted that employs the
regenerative capacity of the body to form a living, autologous
tissue. Ideally, the resulting tissue-engineered construct should
grow, repair and remodel like a native tissue (Mayer, 2001;
Yacoub and Takkenberg, 2005), and therefore require no
follow-up operations. Nevertheless, physiological growth and
remodeling
of
tissue-engineered
constructs
remains
challenging. Several studies reported that constructs became
geometrically unstable after implantation as a result of adverse
tissue growth and remodeling, such as aneurysm formation in
vascular grafts (McAllister et al., 2009; Khosravi et al., 2016), and
leaflet shortening in valvular prostheses (Schmidt et al., 2010;
Driessen-Mol et al., 2014; Reimer et al., 2017). In native tissues,
the outcome of growth and remodeling, either physiological or
pathological, is thought to depend at least partly on the
establishment of a homeostatic mechanical state (Humphrey,
2008; Cyron and Humphrey, 2014; Humphrey et al., 2014),
where one or more mechanical quantities in a tissue are
maintained in equilibrium. Various quantities have been
suggested to define such a mechanical homeostasis in different
tissues, including stress (Taber and Humphrey, 2001; Humphrey
et al., 2009; Valentín et al., 2009), strain (Chaput et al., 2008;
Oomen et al., 2016), strain energy density (Cyron and Humphrey,
2014; Vigliotti et al., 2016), and stiffness (Ferruzzi et al., 2016;
Bellini et al., 2017). The mechanical state of tissue-engineered
constructs arises from the external loading conditions, their
geometry and mechanical properties. We therefore believe that
by
understanding
the
interplay
between
tissue
growth,
remodeling,
the
mechanical
state
of
tissue-engineered
constructs, and different external loading conditions, we are
able to optimize and steer growth and remodeling via the
rational design of scaffold geometry, thus creating long-term
functioning grafts. Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 2.1 Model-Driven Design of Initial Scaffold
Geometry to Induce Anisotropic
Deformation The bioreactor (Figure 1A) features two chambers that are
separated
by
a
deformable
membrane. Tissue-engineered
constructs, mounted in inserts (Figures 1C, D), can be
cultured in the top chamber, while being dynamically loaded
via air pressurization of the bottom chamber. The pressure in the
top chamber is monitored real-time, and constant maximum
pressure is maintained via a custom computer-driven feedback
system (van Kelle et al., 2017). This feedback system adjusts the
air pressurization of the bottom chamber continuously to ensure
the target pressure in the top chamber is achieved. In this
bioreactor system, an axisymmetric insert geometry leads to
isotropic in-plane deformation in the construct. In order to
induce anisotropic deformation within these samples, elliptical
inserts were developed, where the main axes measured 7.5 and
15 mm, respectively (Figure 1C). Circular constructs with a
diameter
of
12 mm
were
used
as
isotropic
controls
(Figure 1D). These dimensions were selected based on finite
element modeling in Abaqus FEA (Dassault Systèmes Simulia,
Providence, RI, United States) of pressurized circular and
elliptical tissue-engineered constructs with varying dimensions. Bioreactor platforms provide a valuable means to study this
interplay under tightly controlled conditions. We recently
developed the Vertigro bioreactor, in which tissue-engineered
cardiovascular constructs can be cultured under dynamic
pressurization to mimic hemodynamic loading (Oomen et al.,
2017; van Kelle et al., 2017). Importantly, the platform also allows
for nondestructive mechanical and geometric characterization of
tissue-engineered constructs during culture, allowing us to February 2022 | Volume 10 | Article 796452 Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 2 Hermans et al. Mechanical Evolution of Tissue-Engineered Constructs FIGURE 1 | (A) The Vertigro bioreactor. (B) Scanning electron microscopy image of the scaffold prior to seeding. (C) Elliptical construct in insert measuring 7.5 mm
(blue) by 15 mm (red). (D) Circular construct in insert with a 12 mm diameter (yellow). FIGURE 1 | (A) The Vertigro bioreactor. (B) Scanning electron microscopy image of the scaffold prior to seeding. (C) Elliptical construct in insert measuring 7.5 mm
(blue) by 15 mm (red). (D) Circular construct in insert with a 12 mm diameter (yellow). bioreactor. (B) Scanning electron microscopy image of the scaffold prior to seeding. (C) Elliptical construct in insert measuring 7.5 mm
Circular construct in insert with a 12 mm diameter (yellow). plane of the construct. 2.1 Model-Driven Design of Initial Scaffold
Geometry to Induce Anisotropic
Deformation The contribution of each
fiber to the total strain energy was described by a non-linear
model (Driessen et al., 2007; Oomen et al., 2018) to account for its
strain-stiffening behavior, where the total strain energy density of
the fibrous component Ψf is the sum of the strain energy densities
of all individual fibers Ψi
f, and: σ 2
J F · zΨ ξ
( )
zC
· FT
(1) (1) with F the deformation gradient tensor, C = FT ·F the right
Cauchy-Green deformation tensor and the Jacobian J = det(F). Ψ
was split in two components, the matrix part Ψm and the fibrous
part Ψf: Ψi
f k1
2k2
exp k2〈λi
2 −1〉
−k2〈λi
2 −1〉−1
(5) (5) Ψ 1 −Φf
Ψm + ΦfΨf
(2) (2) with material parameters k1 and k2, the squared elastic fiber
stretch (λi)2 C: (ei
f0 ⊗ei
f0), and Macaulay brackets (〈◦〉) to
enforce that the fibers only resist tension. The material
parameters were set to resemble the material properties of an
isotropic polycaprolactone-bisurea (PCL-BU) scaffold, which
were
determined
using
a
BioTester
(CellScale,
Waterloo,
Canada): μ = 1.45 ·102 kPa, k1 = 1.32 ·107 kPa, k2 = 2.64 ·10–5,
and β = 100 to enforce an isotropic scaffold fiber distribution. The
bulk modulus was estimated via κ 2μ(1+])
3(1−2]) with shear modulus μ
and an assumed Poisson’s ratio ] of 0.495. with material parameters k1 and k2, the squared elastic fiber
stretch (λi)2 C: (ei
f0 ⊗ei
f0), and Macaulay brackets (〈◦〉) to
enforce that the fibers only resist tension. The material
parameters were set to resemble the material properties of an
isotropic polycaprolactone-bisurea (PCL-BU) scaffold, which
were
determined
using
a
BioTester
(CellScale,
Waterloo,
Canada): μ = 1.45 ·102 kPa, k1 = 1.32 ·107 kPa, k2 = 2.64 ·10–5,
and β = 100 to enforce an isotropic scaffold fiber distribution. The
bulk modulus was estimated via κ 2μ(1+])
3(1−2]) with shear modulus μ
and an assumed Poisson’s ratio ] of 0.495. in which the total fiber volume fraction Φf was set at 0.5 (Driessen
et al., 2007). 2.1 Model-Driven Design of Initial Scaffold
Geometry to Induce Anisotropic
Deformation Each individual fiber (i) was associated
with a vector ei
f0 (equally-spaced in the reference configuration)
that represents its direction within the tissue plane, and a volume
fraction ϕi
f that is described by a Gaussian function: With the selected shapes and dimensions, anisotropic and
isotropic deformation was achieved with similar maximum
deformation and stress. Due to the symmetry of both shapes,
only a quarter of each construct was modeled. Nodes on the outer
edge of the constructs were fully encastered, while symmetry
boundary conditions were applied on the inner edges. A pressure
of 6 kPa was applied from underneath the tissue to simulate
mechanical loading in the bioreactor. The constructs were
modeled using a fiber-reinforced material model (Oomen
et al., 2017), consisting of an isotropic matrix and a fibrous
network. The Cauchy stress (σ) was determined via a strain
energy density function Ψ as a function of a material
parameter set ξ: With the selected shapes and dimensions, anisotropic and
isotropic deformation was achieved with similar maximum
deformation and stress. Due to the symmetry of both shapes,
only a quarter of each construct was modeled. Nodes on the outer
edge of the constructs were fully encastered, while symmetry
boundary conditions were applied on the inner edges. A pressure
of 6 kPa was applied from underneath the tissue to simulate
mechanical loading in the bioreactor. The constructs were
modeled using a fiber-reinforced material model (Oomen
et al., 2017), consisting of an isotropic matrix and a fibrous
network. The Cauchy stress (σ) was determined via a strain
energy density function Ψ as a function of a material
parameter set ξ: ϕi
f A exp −γi −α
2
β2
. (4) (4) Here, the angle of the main fiber direction α is set to 0 (parallel
to the long axis of the elliptical constructs), the angle γi defines the
orientation of a fiber i with respect to α, β represents the fiber
dispersity, and A is a normalization factor to ensure that the sum
of individual fiber fractions equals Φf. 2.3 Monitoring the Evolving Mechanical
State and Geometry of Dynamically Loaded
Constructs
2.3.1 Nondestructive Mechanical and Geometric
Characterization 2.2 Tissue Culture in the Vertigro Bioreactor
In total, 16 circular and 16 elliptical tissue-engineered constructs
were prepared. Bare, 0.3 mm thick electrospun PCL-BU scaffolds
(Wisse et al., 2006) with isotropically distributed fibers (fiber
diameter of approximately 4 μm) (Figure 1B) were cut from a
sheet using a cork borer, mounted in the inserts and sterilized
using UV light. Following sterilization, inserts were submerged in
0.25 mg/mL L-ascorbic 2-phosphate acid (Sigma-Aldrich, St. Louis, MO, United States) supplemented standard culture
medium (TE medium) and placed in an incubator overnight. Human vena saphena cells (HVSCs), which were previously
characterized as contractile, matrix-producing myofibroblasts
(Mol et al., 2005), were isolated from human vena saphena
samples
according
to
the
Dutch
guidelines
for
use
of
secondary materials. These cells were cultured up to passage
seven
in
advanced
Dulbecco’s
Modified
Eagle
Medium
(Invitrogen, Carlsbad, CA, United States), supplemented with
10% Fetal Bovine Serum (Greiner Bio One, Frickenhausen,
Germany), 1% Glutamax (Invitrogen) and 1% penicillin/
streptomycin (Lonza, Basel, Switzerland). After trypsinization,
the cells were seeded in the sterilized PCL-BU scaffolds at a
density of 15 million cells/cm3 using fibrin as a cell carrier (Mol
et al., 2005). To allow tissue formation inside the scaffolds before
dynamic loading, all constructs were statically cultured for
14 days in TE medium in an incubator at 37 °C, 100% relative
humidity and 5% CO2, where medium was changed twice a week. After 14 days of static culture, five elliptical and six circular
constructs
were
sacrificed
for
analysis
of
structure
and
composition. These samples served as day 0 control samples
for the other eight elliptical and circular constructs that were then
dynamically loaded for 14 days in the bioreactor at a peak
pressure of 6 kPa applied at a rate of 1 Hz. After dynamic
loading, the remaining samples were sacrificed as well for
analysis of structure and composition. For an overview of the
experimental design including sample sizes, Figure 2. Throughout the period of dynamic loading, constructs were
mechanically characterized in a nondestructive manner on day
0, 4, 7, 11 and 14 (Figure 2), using a previously developed method
(Oomen et al., 2017; van Kelle et al., 2017). 2.1 Model-Driven Design of Initial Scaffold
Geometry to Induce Anisotropic
Deformation After dynamic loading, the remaining samples (8 elliptical
and seven circular constructs) were also sacrificed for analysis of structure and composition (c. dynamically loaded elliptical constructs, d. dynamically loaded circular
constructs). (B) Overview of the number of mechanically characterized samples per construct geometry during dynamic loading. Numbers vary per day because during
some measurements, pressure data was found to be improperly calibrated. These samples were therefore omitted from the analysis of that particular time point. 2.3 Monitoring the Evolving Mechanical
State and Geometry of Dynamically Loaded
Constructs
2.3.1 Nondestructive Mechanical and Geometric
Characterization Briefly, a 12 MHz
linear ultrasound transducer on a MyLab 70 Ultrasound system
(LA435 probe, Esaote, Maastricht, Netherlands) was placed on
top of the Vertigro bioreactor to image the cross-section of the
construct along the diameter of the circular constructs and both
the short and long axes of the elliptical constructs (Figure 1C). During
ultrasound
imaging,
constructs
were
gradually
pressurized up to 6 kPa in 10 s. Afterwards, ultrasound images
were processed using a custom MATLAB (MathWorks, Natick,
MA, United States) script to obtain the constructs’ cross-sectional
profile as a function of pressure. In case of elliptical constructs,
the profiles along both axes were interpolated by taking the
average height of the apex of each image to obtain one mean
profile. Additionally, construct thickness was determined by
collecting
raw
radiofrequency
data
from
non-pressurized
constructs. Using MATLABs findpeaks algorithm, the local
maxima, or peaks, were identified in the raw data. The two
highest peaks were defined as the tissue borders. Average
construct thickness was estimated based on the distance
between these two peaks in 20 locations per construct. 2.1 Model-Driven Design of Initial Scaffold
Geometry to Induce Anisotropic
Deformation The isotropic matrix part was modeled as a Neo-
Hookean material: Ψm κ
2ln2 J
( ) + μ
2 I1 −3 −2 ln J
( )
(
)
(3) (3) with bulk modulus κ, shear modulus μ and the first invariant of
the right Cauchy-Green deformation tensor I1 = C: I. The fibrous
component was modeled with a discrete number of fibers in the February 2022 | Volume 10 | Article 796452 Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 3 Mechanical Evolution of Tissue-Engineered Constructs Hermans et al. FIGURE 2 | (A) Overview of the experimental design. All samples (n = 16 for both geometries) underwent static culture for 14 days. During static culture, three
elliptical and two circular constructs detached from the insert and were therefore omitted from the analyses. After static culture, five elliptical and six circular statically
cultured constructs were sacrificed for analysis of structure and composition (a. statically cultured elliptical constructs, b. statically cultured circular constructs). The
remaining eight constructs per geometry were dynamically loaded for 14 days. During these 14 days, constructs were mechanically characterized on day 0, 4, 7,
11, and 14. During dynamic loading, one circular construct detached from the insert and was omitted as well. After dynamic loading, the remaining samples (8 elliptical
and seven circular constructs) were also sacrificed for analysis of structure and composition (c. dynamically loaded elliptical constructs, d. dynamically loaded circular
constructs). (B) Overview of the number of mechanically characterized samples per construct geometry during dynamic loading. Numbers vary per day because during
some measurements, pressure data was found to be improperly calibrated. These samples were therefore omitted from the analysis of that particular time point. FIGURE 2 | (A) Overview of the experimental design. All samples (n = 16 for both geometries) underwent static culture for 14 days. During static culture, three
elliptical and two circular constructs detached from the insert and were therefore omitted from the analyses. After static culture, five elliptical and six circular statically
cultured constructs were sacrificed for analysis of structure and composition (a. statically cultured elliptical constructs, b. statically cultured circular constructs). The
remaining eight constructs per geometry were dynamically loaded for 14 days. During these 14 days, constructs were mechanically characterized on day 0, 4, 7,
11, and 14. During dynamic loading, one circular construct detached from the insert and was omitted as well. 2.3.2 Estimation of Material Properties and
Mechanical State Using the construct’s cross-sectional profile as a function of
pressure (from 0 to 6 kPa), the material properties were
estimated via a two-step inverse method as described in February 2022 | Volume 10 | Article 796452 Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 4 Mechanical Evolution of Tissue-Engineered Constructs Hermans et al. Oomen et al., 2017. In brief: as a first step, an initial estimate of the
material properties was made based on shell theory [see (Oomen
et al., 2017) for further details] and the construct’s cross-sectional
profile as a function of pressure. In the second step, a full inverse
finite element method was employed to obtain a more accurate
estimate. Constructs were modeled using the same fiber-
reinforced material model as described in Section 2.1. determined using a Qubit 2.0 fluorometer (Invitrogen). Lastly,
a quarter of each construct was analyzed using the Fastin Elastin
assay kit (Fastin, Biocolor) to quantify elastin content, which was
normalized to construct wet weight. 2.5.2 In-Plane Collagen Orientation g
The remaining part of each construct was analyzed for the in-
plane collagen fiber orientation. Firstly, each sample was stained
for collagen using a fluorescent CNA35 probe (1 h incubation
time) (Boerboom et al., 2007). Secondly, tile scans (excitation
488 nm, emission 520 nm, magnification ×10, maximum 60 μm
deep) of both the top and bottom side of stained samples were
made using a confocal laser scanning microscope (TCS SP5X;
Leica
Microsystems,
Wetzlar,
Germany). Collagen
fiber
distributions were quantified in MATLAB (Frangi et al., 1998). In brief, tissue structure was analyzed based on the Hessian
matrix. Fiber-like structures were identified by means of the
Frangi Vesselness. The principal direction of a fiber was
subsequently determined by decomposition of the Hessian
matrix. In this way, a histogram with the fraction of collagen
fibers oriented in each in-plane direction was obtained, which was
parameterized by means of a normalized unimodal Gaussian
distribution using previously described algorithms (van Haaften
et al., 2018). This algorithm provided an estimate for the fraction
of aligned fibers in the whole construct. Following estimation of the material parameters in each
timepoint, the Cauchy stress tensor, deformation gradient
tensor, and the strain energy density at peak pressure were
obtained from the nodes in the central 50% of the samples. Next, the principal stresses σi and their directions ni were
determined (with i = 1, 2, 3) by calculating the eigenvalues
and eigenvectors, respectively, of the Cauchy stress tensor. The
principal stretches λi were obtained from λi 1/
B−1: (ni ⊗ni)
,
with i = 1, 2, 3 and B = F ·FT the left Cauchy-Green deformation
tensor. The deformation gradient tensor F was provided by the
finite element model. The maximum principal stress and stretch
were analyzed for the circular constructs, while for the elliptical
constructs the maximum and middle principal stress and stretch
were found to approximately correspond with the in-plane stress
and stretch along the short and long axis, respectively. The local
tangent of the principal stress-stretch curves was determined at
the peak pressure of 6 kPa. To quantify the overall mechanical
state of a construct, these quantities were averaged for the inner
50% of each construct. 2.5 Analysis of Tissue Structure
2.5.1 Histology From the remaining quarter of each construct, the two straight
rims were cut off, thus obtaining an L-shaped part that coincides
with the two semi-axes of the whole construct. This part was
embedded in paraffin and sectioned through its thickness into
slices of 7 μm. For the elliptical samples, sections were made along
both the short and long axes. Part of these embedded sections
were stained for general tissue structure [Hematoxylin and Eosin
(HE)], and collagen [Picrosirius red (PR)], and subsequently
imaged using a brightfield microscope (Axio Observer Z1, Carl
Zeiss AG, Oberkochen, Germany). On the remaining sections,
immunohistochemistry was performed to visualize (tropo)
elastin,
alpha-smooth
muscle
actin
(α-SMA),
as
well
as
collagen type I and III. These sections were imaged using an
Axiovert 200M microscope (Zeiss). In the second step, the final material parameters were
estimated using MATLAB’s lsqnonlin function, starting from
the
initial
estimates
of
the
first
step. This
function
incrementally adjusts the material parameter set [ξ = (μ, k1, k2,
β)], compares the simulated cross-sectional profile of the
construct with the construct cross-sectional profile obtained
from the nondestructive mechanical testing, and repeats this
process until the error E between these profiles, as a function
of pressure (p), was minimized: E ξ
( )
1
NpNx
Np
j1
Nx
k1
z pj, xk, ξ
exp −z pj, xk, ξ
est
(6) (6) where Np is the number of pressure increments, and Nx the
number of in-plane positions x along the construct profile. The
inclusion of β in the material parameter set enables the model to
account for mechanical anisotropy that may occur as a result of
structural remodeling in the engineered constructs (Eq. (4)). 2.6 Statistics Numerical data are presented as mean ± standard error of the
mean. The longitudinal geometric and mechanical data were
analyzed in RStudio (RStudio Team, Boston, United States)
using a mixed linear effects model, which accounts for
correlations between the repeated measurements and remains
reliable in case of samples missing completely at random
(Hothorn and Everitt, 2009), as is the case here. Post hoc
analysis to test for significant differences between groups at
the same timepoint and between timepoints within a single
group was done using the Estimated Marginal Means function. Biochemical assay data between groups was compared with the
multiple comparison procedure developed by Herberich et al. (2010) in R 3.5.0 (The R foundation, Vienna, Austria). With this 3.2 Different Patterns of Enlargement
Depending on Initial Scaffold Geometry At the start of the culture period, elliptical constructs measured
7.5 and 15 mm along the short and long axis respectively, and
the circular constructs 12 mm in diameter (L0). Constructs
enlarged during dynamic loading (Figure 3A), which was
confirmed
by
the
ultrasound
data
of
a
representative
(circular) construct at zero pressure right before (Figure 3C)
and on the last day of dynamic loading (Figure 3D). Elliptical
constructs enlarged unevenly, with higher elongation along
their short axis compared to their long axis (Figure 3A). The
elongation along the elliptical constructs’ short axis and the
cross-section of the circular constructs was similar until day 7,
after which the elliptical constructs enlarged more than the 3.3 Different Initial Scaffold Geometries
Lead to Distinct Mechanical States The selected elliptical geometry resulted in stretch anisotropy,
with higher stretch along the short compared to the long axis
(Supplementary Figure S1 left and Supplementary Figure S2). As expected, the in-plane distribution of stretch was isotropic in
the circular construct when pressurized (Supplementary Figure
S1 right). Importantly, these designs resulted in similar stress-
stretch at maximum pressure along the short axis of the elliptical
construct and the circular construct (Supplementary Figure S2). Using nondestructive mechanical testing and inverse finite element
analysis, we monitored the evolution of four mechanical quantities in
the elliptical and circular constructs (Figure 4). Interestingly, different
mechanical states were observed between constructs, depending on
the initial scaffold geometry. On day 0 of dynamic loading, all
quantities except the strain energy density were already at different
levels between the different construct geometries. After 4 days of
dynamic loading, a stretch and strain energy density equilibrium was
established and maintained in the elliptical constructs, whereas these
quantities continuously increased in the circular constructs during
dynamic loading (Figures 4A, D). The tangent stiffness (Figure 4B)
and the maximum/middle principal Cauchy stress (Figure 4C), were
at a higher level in the circular compared to the elliptical constructs,
and did not change over time in neither construct type. 2.4 Analysis of Tissue Composition y
p
After sacrificing a construct, one half was snap-frozen in liquid
nitrogen, lyophilized overnight and pulverized using a Mikro-
Dismembrator (Model S, Sartorius, Goettingen, Germany). The
remains were digested for 16 h at 60°C using a papain digestion
buffer. The amount of GAGs in the digest was quantified using an
adapted protocol by Farndale et al. (1986), with a standard curve
of chondroitin sulfate (Shark Cartilage, Sigma-Aldrich). Using a
trans-4-hydroxyproline (Sigma-Aldrich) standard curve, the
hydroxyproline (HYP) content (as a measure for collagen
content) was quantified in accordance with the protocol of
Huszar et al. (1980). Additionally, the DNA content was February 2022 | Volume 10 | Article 796452 Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 5 Mechanical Evolution of Tissue-Engineered Constructs Hermans et al. FIGURE 3 | (A) Relative changes in cross-sectional length at zero pressure with respect to the initial length at the start of the experiment (L/L0) of the short (blue) and
long (red) axis of the elliptical constructs and the diameter of the circular constructs (yellow), as well as (B) construct thickness during 14 days of dynamic loading. See
supplemental material for p-values. (C) Ultrasound image of a circular construct at the start of dynamic loading, and (D) a dilated circular construct at the end of dynamic
loading at zero pressure (p = 0). FIGURE 3 | (A) Relative changes in cross-sectional length at zero pressure with respect to the initial length at the start of the experiment (L/L0) of the short (blue) and
long (red) axis of the elliptical constructs and the diameter of the circular constructs (yellow), as well as (B) construct thickness during 14 days of dynamic loading. See
supplemental material for p-values. (C) Ultrasound image of a circular construct at the start of dynamic loading, and (D) a dilated circular construct at the end of dynamic
loading at zero pressure (p = 0). circular constructs. Interestingly, the length of the cross-section
of the circular constructs stabilized from day 11. procedure,
no
assumptions
are
necessary
regarding
the
distribution, sample sizes or variance homogeneity. Differences
were considered significant if p ≤0.05. On day 0 of dynamic loading, elliptical constructs were slightly
thicker than the circular constructs. From day 4, elliptical
construct thickness decreased and remained similar to circular
construct thickness for the rest of dynamic loading (Figure 3B). 3.4 Initial Scaffold Geometry Influences
Construct Elastin Content The
composition
of
statically
and
dynamically
cultured
constructs was quantified in terms of GAG, DNA, collagen
(HYP), and (tropo)elastin content. There were no differences
in the elliptical and circular constructs with respect to DNA
content, GAG per DNA, and Hyp per DNA, both before and after February 2022 | Volume 10 | Article 796452 Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 6 Mechanical Evolution of Tissue-Engineered Constructs Hermans et al. FIGURE 4 | Evolution of the principal elastic stretch (A), tangent stiffness of the principal stress-stretch curve (B), principal Cauchy stress (C), and strain energy
density (D) at 6 kPa in the central 50% of elliptical and circular constructs during dynamic loading. In (A–C) the maximum and middle principal component for the elliptical
constructs and the maximum component for the circular constructs are given. In the ellipses, the maximum and middle principal components of the Cauchy stress and
stretch are directed along the short and long axis approximately. See supplemental material for p-values. FIGURE 4 | Evolution of the principal elastic stretch (A), tangent stiffness of the principal stress-stretch curve (B), principal Cauchy stress (C), and strain energy
density (D) at 6 kPa in the central 50% of elliptical and circular constructs during dynamic loading. In (A–C) the maximum and middle principal component for the elliptical
constructs and the maximum component for the circular constructs are given. In the ellipses, the maximum and middle principal components of the Cauchy stress and
stretch are directed along the short and long axis approximately. See supplemental material for p-values. FIGURE 5 | Average (A) DNA content, (B) GAG per DNA, (C) HYP per DNA, and (D) (tropo)elastin per wet weight in elliptical (E) and circular (C) constructs before
(d0) and after 14 days (d14) of dynamic loading. One asterisk (*) represents p ≤0.05, two p ≤0.01, and three p ≤0.001. FIGURE 5 | Average (A) DNA content, (B) GAG per DNA, (C) HYP per DNA, and (D) (tropo)elastin per wet weight in elliptical (E) and circular (C) constructs before
(d0) and after 14 days (d14) of dynamic loading. One asterisk (*) represents p ≤0.05, two p ≤0.01, and three p ≤0.001. dynamic loading (Figure 5). During dynamic loading, DNA
content increased and GAG and Hyp per DNA decreased in
all constructs in similar proportions, implying that collagen and
GAG contents remained similar and cell numbers increased
during dynamic loading. 3.4 Initial Scaffold Geometry Influences
Construct Elastin Content On the other hand, (tropo)elastin per
construct wet weight was higher in the elliptical than the circular
constructs, and remained similar during dynamic loading. dynamic loading (Figure 5). During dynamic loading, DNA
content increased and GAG and Hyp per DNA decreased in
all constructs in similar proportions, implying that collagen and
GAG contents remained similar and cell numbers increased
during dynamic loading. On the other hand, (tropo)elastin per
construct wet weight was higher in the elliptical than the circular
constructs, and remained similar during dynamic loading. 3.5 Anisotropic Deformation Results in
Anisotropic In-Plane Collagen Organization
Histology revealed that the elliptical and circular constructs were
structurally similar. Additionally, the structure Along the short
and long axis of the elliptical constructs were similar as well. The
HE staining showed that the HVSCs were homogeneously
distributed
throughout
the
thickness
of
all
constructs Frontiers in Bioengineering and Biotechnology | www.frontiersin.org February 2022 | Volume 10 | Article 796452 7 Mechanical Evolution of Tissue-Engineered Constructs Hermans et al. FIGURE 6 | Representative histological (A) and immunohistochemical (B) cross-sections along the short and long axis of the elliptical constructs, and the diameter
of the circular samples, before (day 0) and after (day 14) dynamic loading. Note that the bottom side of all samples coincides with the pressurized side in the bioreactor. Scale bars represent 100 μm. (A) The top two rows show a Hematoxylin and Eosin staining to indicate the general tissue composition and cellular distribution. The
bottom two rows feature a PR stain, depicting collagen fibers (red). (B) From top to bottom; elastin (red), collagen type 3 (red), collagen type 1 (red) and α-SMA
(green). In all images cell nuclei are stained blue. Negative controls for immunohistochemical images can be found in Supplementary Figure S3. FIGURE 6 | Representative histological (A) and immunohistochemical (B) cross-sections along the short and long axis of the elliptical constructs, and the diameter
of the circular samples, before (day 0) and after (day 14) dynamic loading. Note that the bottom side of all samples coincides with the pressurized side in the bioreactor. FIGURE 6 | Representative histological (A) and immunohistochemical (B) cross-sections along the short and long axis of the elliptical constructs, and the diameter
of the circular samples, before (day 0) and after (day 14) dynamic loading. Note that the bottom side of all samples coincides with the pressurized side in the bioreactor. Scale bars represent 100 μm. Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 3.4 Initial Scaffold Geometry Influences
Construct Elastin Content (A) The top two rows show a Hematoxylin and Eosin staining to indicate the general tissue composition and cellular distribution. The
bottom two rows feature a PR stain, depicting collagen fibers (red). (B) From top to bottom; elastin (red), collagen type 3 (red), collagen type 1 (red) and α-SMA
(green). In all images cell nuclei are stained blue. Negative controls for immunohistochemical images can be found in Supplementary Figure S3. (Figure 6A). In both the HE and PR images, a dense tissue layer
was visible on the top side of all statically cultured samples. Interestingly, after dynamic loading this layer was found on the
opposite, pressurized side (Figure 6A). on the in-plane structural collagen organization (Figure 7). Before dynamic loading (day 0), the fraction of anisotropic
collagen fibers was similar in the circular and elliptical
constructs (day 0) (Figures 7A,B,E). After dynamic loading
(day 14), the average anisotropic fiber fraction was increased
for the elliptical constructs whereas it remained similar for the
circular constructs (Figures 7C–E). The presence of a small
anisotropic collagen fiber fraction in the day 0 constructs and
the circular day 14 constructs was also seen in the scaffold fibers
(Supplementary Figure S4). The preferred orientations of the
collagen and scaffold fibers in these samples coincided, suggesting
that the scaffold fibers provided contact guidance cues to the cells
that then produced collagen along their long axis, resulting in a
collagen organization matching with the scaffold fiber directions. In the statically cultured constructs, both tropoelastin
aggregates and collagen type III were primarily found at the
top and bottom surfaces, whereas after dynamic loading these
were seen across the whole thickness (Figure 6B). A dense
collagen type I layer was observed on top of the constructs
after static culture, which after dynamic loading was present
on the bottom instead. Only a few α-SMA-positive cells were
found in the constructs, regardless of construct geometry and
culture time (Figure 6B). On the other hand, the initial construct
geometry in combination with dynamic loading had a clear effect February 2022 | Volume 10 | Article 796452 Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 8 Mechanical Evolution of Tissue-Engineered Constructs Hermans et al. FIGURE 7 | The top and bottom side of representative collagen-stained quarters of elliptical (A,C) and circular (B,D) constructs before (day 0) (A,B) and after (day
14) (C,D) dynamic loading. 3.4 Initial Scaffold Geometry Influences
Construct Elastin Content The histograms indicate the fraction of collagen fibers oriented in each direction for each image. Within histograms, the green line represents a
Gaussian fit for the fiber distribution. The fiber fraction above the red baseline represents the anisotropic fiber fraction based on the Gaussian fit. (E) Average anisotropic
fiber fraction of the top and bottom side of the pre-dynamic loading (d0) elliptical (E) and circular (C) constructs (both n = 2), and the post-dynamic loading (d14)
elliptical and circular constructs (both n = 5). FIGURE 7 | The top and bottom side of representative collagen-stained quarters of elliptical (A,C) and circular (B,D) constructs before (day 0) (A,B) and after (day
14) (C,D) dynamic loading. The histograms indicate the fraction of collagen fibers oriented in each direction for each image. Within histograms, the green line represents a
Gaussian fit for the fiber distribution. The fiber fraction above the red baseline represents the anisotropic fiber fraction based on the Gaussian fit. (E) Average anisotropic
fiber fraction of the top and bottom side of the pre-dynamic loading (d0) elliptical (E) and circular (C) constructs (both n = 2), and the post-dynamic loading (d14)
elliptical and circular constructs (both n = 5). 4 DISCUSSION dynamic loading in vitro. Besides distinct mechanical states, we also
observed differences in the final collagen organization of constructs
depending on the initial scaffold geometry. As the biomechanical
behavior
of
cardiovascular
tissues
effectively
defines
their
functionality, these findings indicate that initial scaffold geometry
can determine the short-term functionality of cardiovascular tissue-
engineered constructs while the scaffold is still present in the
construct. Additionally, all constructs dilated during dynamic
loading regardless of initial scaffold geometry, a property that
must be considered in the in vivo application of PCL-BU
scaffolds, as ultimately the functionality of many cardiovascular
tissues such as heart valves and blood vessels relies on their
dimensions. In situ tissue engineering relies on growth and remodeling to build a
functional
tissue
from
a
non-living
scaffold. Therefore,
understanding, predicting, and directing growth and remodeling in
tissue-engineered constructs is key to enable successful in situ tissue
engineering strategies. To obtain control over growth and remodeling,
we must explore how scaffold parameters, such as geometry, influence
the interplay between tissue growth, remodeling, and the construct’s
mechanical state, thereby ultimately enabling the rational design of
scaffolds that promote functional growth and remodeling. To this
end, here we demonstrated that initial scaffold geometry dictates the
mechanical state of tissue-engineered constructs during short-term February 2022 | Volume 10 | Article 796452 Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 9 Mechanical Evolution of Tissue-Engineered Constructs Hermans et al. long axis were required at each time point. The data from both
axes were interpolated, which added an uncertainty to the
geometric
and
mechanical
data
of
these
constructs. Theoretically, the apex displacement from both measurements
should be identical, although in practice this was not always
exactly the case. This can be attributed to the fact that the
ultrasound
probe
was
manually
positioned
during
the
ultrasound measurements, and therefore was not perfectly
aligned with the short and long axis. An effective solution to
this problem would be the use of 3D echography, which allows
the entire construct to be imaged at once, omitting the necessity
of interpolation. Additionally, collagen orientation could only be
quantified up to 60 μm deep using confocal microscopy. As a
solution, high frequency ultrasound imaging could be used to
evaluate the collagen organization throughout the construct
thickness, as performed by Riggin et al. (2014) in their study
on rat tendons in vivo. 4 DISCUSSION Applying this technique in the current
study would help to gain valuable insights into the structural
collagen fiber adaptation process. Despite accounting for the
different volumes of the elliptical and circular scaffolds at the
start of culture by seeding constructs with the same cell density,
the measured DNA content at end of culture was similar for the
elliptical and circular constructs (Figure 5A) albeit the circular
constructs had an ≈1.3 times larger volume. This implies that at
the end of culture the cell density was ≈1.3 times higher in the
elliptical constructs, possibly due to a difference in cell
proliferation depending on the geometry of the scaffold. This
could also explain the factor ≈1.3 higher density of elastin in the
elliptical compared to the circular constructs (Figure 5D),
assuming all cells produced a similar amount of elastin. Lastly,
our engineered constructs displayed viscoelastic behavior and
permanent deformation, while we only incorporated elastic
behavior in our material model. To account for this limitation
at least partially, we determined construct profiles non-invasively
in reference state (at p = 0 kPa) and loaded state (at p = 6 kPa) for
each
individual
construct
on
every
day
of
experimental
characterization, and incorporated these changes in reference
configuration in our computational analyses. Additionally, we
mechanically tested constructs with a single load cycle from 0 to
6 kPa in 10 s to minimize the influence of viscoelastic behavior. Our findings are in agreement with previous studies that
reported distinct mechanical and geometric states in tissue-
engineered constructs depending on initial scaffold thickness,
both in vitro (van Kelle et al., 2018) and in vivo (Wu et al., 2021). Even microscale geometric properties, such as fiber alignment
affect the short-term mechanical evolution of engineered
constructs (Uiterwijk et al., 2020). Collectively, these results
show that construct functionality is highly dependent on
scaffold design. g
We observed different mechanical equilibria in our constructs
depending on the initial scaffold geometry. Additionally, the
circular constructs appeared to reach geometric stability (i.e. equilibria in terms of thickness and in-plane dimensions), in
contrast to the elliptical constructs. However, it remains
uncertain to what extent these equilibria are a manifestation of
a homeostasis that is established by the cells in these constructs,
or whether the cells have little to no influence on the behavior of
the construct due to the presence of the PCL-BU scaffold. 4 DISCUSSION We
hypothesize that the latter is the case as there are no structural or
compositional differences between the elliptical and circular
constructs that clearly explain the different mechanical states. Possibly, the differences in evolution of the mechanical state can
be explained by the initial scaffold geometry and permanent
deformation that is known to occur in these scaffolds (van Kelle
et al., 2018). Furthermore, it remains an open question if, once the
scaffold starts to degrade, the observed mechanical and geometric
equilibria would be maintained over time. Alternatively, the lack
of mechanical stability (i.e. equilibria in terms of (all) mechanical
quantities) or geometric stability in the circular and elliptical
constructs, respectively, could be interpreted as a hallmark of
adverse growth and remodeling. However, Drews and colleagues
predicted computationally and showed experimentally that early
stenosis can be spontaneously resolved in tissue-engineered
vascular grafts in vivo (Drews et al., 2020). From this
perspective, it could well be possible that on longer timescales
the initial enlargement of our elliptical and circular constructs is
reversed, and stretch and strain energy density equilibria are
established in our circular constructs. For future research, it would be interesting to monitor the
mechanical
state
of
constructs
with
degradable
scaffolds,
considering the profound effects that scaffold degradation can
have on both collagen organization and the mechanical state of
tissue-engineered constructs (de Jonge et al., 2014; Uiterwijk et al.,
2020). Additionally, in our pressure-controlled bioreactor, the PCL-
BU scaffolds could undergo unrestricted permanent deformation,
which continuously altered the reference configuration of the
constructs, and therefore could have influenced mechanically-
driven tissue growth and remodeling through the impact that
the reference configuration has on the obtained stresses and
strains. Finally, the role of the immune system in matrix
production and scaffold degradation in situ tissue-engineeing
could be addressed in future work, for instance by including
macrophages (Wissing et al., 2017) in tissue culture. Ultimately,
the success of in situ tissue engineering strategies will depend on the
interplay of all these factors. REFERENCES Driessen-Mol, A., Emmert, M. Y., Dijkman, P. E., Frese, L., Sanders, B., Weber, B.,
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Tissue, Cellular, and Sub-cellular Levels. ETHICS STATEMENT Ethical review and approval was not required for the study on
human participants in accordance with the local legislation and
institutional requirements. The patients/participants provided
their written informed consent to participate in this study. AUTHOR CONTRIBUTIONS SL and CB conceptualized the study. LH conducted the
experiments, the numerical simulations, and analyzed the data. PO and MvK oversaw experiments and simulations. LH and MvK
wrote the manuscript. RL was involved in analyzing and
interpreting the ultrasound data. SL and CB supervised the
project. All authors reviewed the manuscript. 5 CONCLUSION In summary, we studied how anisotropic versus isotropic mechanical
loading—as imposed by initial scaffold geometry—affects tissue
growth and remodeling, and the evolution of the mechanical and
geometric state in cardiovascular tissue-engineered constructs over
time. Using a bioreactor that allows culturing of tissue-engineered
constructs in combination with nondestructive mechanical testing,
we found that initial scaffold geometry dictates how the mechanical
state, geometry, and structural organization of tissue-engineered
cardiovascular
constructs
evolve
in
the
early
phase
of
regeneration. These results demonstrate that mediating the initial
mechanical state of tissue-engineered constructs via scaffold
geometry might be an interesting approach to influence tissue
growth and remodeling and thus modulate tissue outcomes. There are several limitations to this study. For the elliptical
constructs, two ultrasound measurements along the short and February 2022 | Volume 10 | Article 796452 Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 10 Mechanical Evolution of Tissue-Engineered Constructs Hermans et al. DATA AVAILABILITY STATEMENT The authors acknowledge Koo Rijpkema for his advice on the
statistical methods, Marloes Janssen-van den Broek for her advice
on the histological analysis, Leda Klouda for reviewing the
manuscript and Wojciech Szymczyk for electrospinning the
scaffolds. We also want to acknowledge Jurgen Bulsink for
designing
and
manufacturing
the
bioreactor
inserts. We
acknowledge the funding from the European Union’s Seventh
Framework Programme (Grant Agreement No 604 514), and the
Netherlands Cardiovascular Research Initiative (CVON 2012-
01): The Dutch Heart Foundation, Dutch Federation of
University Medical Centres, the Netherlands Organization for
Health Research and Development and the Royal Netherlands
Academy of Sciences. The raw data supporting the conclusion of this article will be
made available by the authors, without undue reservation. SUPPLEMENTARY MATERIAL The Supplementary Material for this article can be found online at:
https://www.frontiersin.org/articles/10.3389/fbioe.2022.796452/
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All rights reserved. No part of this publication may be reproduced, stored in a
retrieval system or transmitted in any form or by any means, electronic, mechanical,
photocopying or otherwise, without the prior permission of the publisher. Negotiating the Sacred II
Blasphemy and Sacrilege in the Arts MARIA SUZETTE FERNANDES-DIAS (Editors) Published by ANU E Press
The Australian National University
Canberra ACT 0200, Australia
Email: anuepress@anu.edu.au
This title is also available online at: http://epress.anu.edu.au/nts02 _citation.html National Library of Australia
Cataloguing-in-Publication entry
Title:
Negotiating the sacred 2 : blasphemy and sacrilege in the
arts / editors, Elizabeth Burns Coleman ; Maria Suzette
Fernandes-Dias. ISBN:
9781921536267 (pbk.)
9781921536274 (pdf)
Subjects:
Arts and religion. Offenses against religion. Blasphemy. Sacrilege. Religion and sociology. Other Authors/Contributors:
Coleman, Elizabeth Burns, 1961-
Fernandes-Dias, Maria Suzette. Dewey Number: 306.108 Dewey Number: 306.108 All rights reserved. No part of this publication may be reproduced, stored in a
retrieval system or transmitted in any form or by any means, electronic, mechanical,
photocopying or otherwise, without the prior permission of the publisher. Cover design by ANU E Press Cover photo by Lynne Broughton Printed by University Printing Services, ANU This edition © 2008 ANU E Press This edition © 2008 ANU E Press Table of Contents Contributors
vii
xi
Acknowledgements
Introduction: Lines in the sand
1
Elizabeth Burns Coleman and Maria Suzette Fernandes-Dias
Section I: Understanding Blasphemy and Sacrilege
9
1. Blasphemy and sacrilege: A challenge to secularisation and theories
11
of the modern? David Nash
2. ‘The devil’s centres of operation’: English theatre and the charge
23
of blasphemy, 1698-1708
David Manning
3. Madonna and piano accordion: Disrupting the order of the world
37
Elizabeth Burns Coleman
4. Materialising the sacred
55
Dianne McGowan
Section II: Motivations for Artistic Blasphemy
67
5. Blasphemy and sacrilege in the novel of magic realism: Grass,
69
Bulgakov, and Rushdie
Peter Arnds
6. Les fees ont soif: Feminist, iconoclastic or blasphemous? 81
Maria-Suzette Fernandes-Dias
7. The body of Christ: Blasphemy as a necessary transgression? 93
Carolyn D’Cruz and Glenn D’Cruz
Section III: Reinterpreting Freedom of Expression
103
8. The monologue of liberalism and its imagination of the sacred in
105
minority cultures
Jasdev Singh Rai
9. Blasphemy in a pluralistic society
127
Jeremy Shearmur Contributors
Acknowledgements Introduction: Lines in the sand
1
Elizabeth Burns Coleman and Maria Suzette Fernandes-Dias Section II: Motivations for Artistic Blasphemy Section II: Motivations for Artistic Blasphemy
67
5. Blasphemy and sacrilege in the novel of magic realism: Grass,
69
Bulgakov, and Rushdie
Peter Arnds
6. Les fees ont soif: Feminist, iconoclastic or blasphemous? 81
Maria-Suzette Fernandes-Dias
7. The body of Christ: Blasphemy as a necessary transgression? 93
Carolyn D’Cruz and Glenn D’Cruz
Section III: Reinterpreting Freedom of Expression
103
8. The monologue of liberalism and its imagination of the sacred in
105
minority cultures
Jasdev Singh Rai
9. Blasphemy in a pluralistic society
127
Jeremy Shearmur Negotiating the Sacred II Section IV: Self-expression and Restriction
145
10. Blasphemy and the art of the political and devotional
147
Christopher Braddock
11. Negotiating the sacred body in Iranian cinema(s): National,
161
physical and cinematic embodiment in Majid Majidi’s Baran (2002)
Michelle Langford
12. Silence as a way of knowing in Yolngu Indigenous Australian
173
storytelling
Caroline Josephs
Bibliography
191
Index
205 Professor Peter Arnds Peter Arnds teaches comparative literature and literary translation, as well as
German and Italian at Trinity College Dublin. He is the author of two books,
one on Wilhelm Raabe and Charles Dickens, and Representation, Subversion, and
Eugenics in Günter Grass’s The Tin Drum. He has published numerous articles
on comparative literature from the eighteenth to the twenty-first century as well
as on German and migrant literature. Section IV: Self-expression and Restriction vi Dr Christopher Braddock Chris Braddock is an artist and academic. He is Associate Professor of Visual
Arts in the School of Art and Design at Auckland University of Technology and
Chair of the University’s St Paul St Gallery. His research interests span the
disciplines of art history, anthropology, and performance studies. He specialises
in different aspects of modern and contemporary art including the part object
in sculpture, performance, body art, and art and spirituality. In 2007 he was the
International Artist-in-Residence at Melbourne’s RMIT University and in 2004
he was the Arts Council of New Zealand toi Aotearoa Visual Arts Resident in
New York City on the International Studio and Curatorial Program. Dr Elizabeth Burns Coleman Elizabeth Burns Coleman is a postdoctoral fellow in the Schools of English,
Communications and Performance Studies and Philosophy and Bioethics at
Monash University. She holds a PhD in philosophy from The Australian National
University and has held a postdoctoral fellowship at the ANU's Centre for Cross
Cultural Research. She has taught philosophy of art at the ANU, moral and
political philosophy at La Trobe University, and philosophy of law at Wollongong
University. She is the author of Aboriginal Art, Identity and Appropriation (2005),
editor (with Kevin White) of Negotiating the Sacred: Blasphemy and Sacrilege in
a Multicultural Society (2006) and has published numerous articles on indigenous
art and cross cultural aesthetics. Her articles have appeared in the Journal of
Aesthetics and Art Criticism, the International Year Book of Aesthetics, and the
Journal of Political Philosophy. Dr Caroline Josephs Caroline Josephs is a free-lance educator, researcher, storyteller, writer, and
artist. Her doctoral thesis was on sacred oral storytelling from four cultures
(Yolngu Indigenous Australian, Zen Buddhist, Judaic, and Inuit). Dr Michelle Langford
Michelle Langford is a lecturer in Film Studies in the School of English, Media
and Performing Arts at the University of New South Wales. She is the author
of Allegorical Images: Tableau, Time and Gesture in the Cinema of Werner Schroeter
(Intellect, 2006) and has published on Iranian cinema in the journal Camera
Obscura. Her current research focuses on the allegorical dimensions of Iranian
cinema. Dr Maria Suzette Fernandez-Dias Dr Maria Suzette Fernandes-Dias is Assistant Director of UN, International and
Regional Organisations Section of the Commonwealth Department of Immigration
and Citizenship. Previously she coordinated scholarly and Research activities
at the Centre for Cross-Cultural Research, ANU. She holds a PhD in French
(postcolonial literature) from the University of Goa, India. She has taught
comparative literature, linguistics and francophone literature at the University
of Goa, and has worked as the educational and cultural coordinator of Alliance
Française de Goa, where she managed the AF Art Gallery. She has an edited
collection, Legacies of Slavery: Comparative Perspectives, (Cambridge Scholars
Publishing, 2007). Her literary awards include the Victor-Hugo Bicentenary
Award (2002), Ford Foundation Campus Diversity Award (1996), OHeraldo
Award for Children's literature (1989), and the Vidya Award (1995 and 1996). Dr Carolyn D’Cruz y
Carolyn D'Cruz is a lecturer in Gender Sexuality and Diversity Studies at La
Trobe University. She is the author of Identity Politics in Deconstruction:
Calculating with the Incalculable (Ashgate, 2008) and has published in
Contretemps: An Online Journal of Philosophy, Life Writing and Social Semiotics. Dr Glenn D’Cruz Dr Glenn D’Cruz
Glenn D'Cruz is Senior Lecturer in drama, media and communication at Deakin
University. He is the author of Midnight's Orphans: Anglo-Indians in Post/Colonial
Literature (Peter Lang, 2006) and Class Act: Melbourne Workers Theatre 1987-2007
(Vulgar Press, 2007). vii Negotiating the Sacred II Dr Jasdev Singh Rai g
Jasdev Singh Rai is a medical doctor and spokesperson for the UK-based Sikh
Human Rights Group. The organisation is based on the humanitarian aspects of
Sikh philosophy: pluralism, non-discrimination, human dignity and bio-ecology. The group has branches in India, Canada, and the USA. The Sikh Human Rights
Group is engaged in community cohesion and diversity issues in the UK. It
conducts projects on transnationalism and human rights among other issues. Dianne McGowan Dianne McGowan is currently completing a PhD at the ANU. She holds two
undergraduate degrees from the ANU: a BSc (psychology major) and a BA
(anthropology major). Between graduating and her PhD research, she established
and managed an Asian art gallery in Sydney, Beowulf Galleries. She is published
in Negotiating the Sacred: Blasphemy and Sacrilege in a Multicultural Society
(Coleman and White (eds.) ANU E Press, 2006). David Manning David Manning is in the final stages of his PhD in History at Clare College,
Cambridge. In a departure from the current historiography of blasphemy, his
dissertation, ‘Blasphemy in England, c.1660-1730’, develops a cultural history
of blasphemy as representation, exploring the nexus between conceptions and
perceived manifestations of blasphemy in a theological context. In 2007 he was
awarded second prize in the annual Pollard Prize of the Institute of Historical
Research, University of London. viii Contributors Dr David Nash David Nash is Reader in History, Oxford Brookes University, UK. He is the author
of Secularism, Art and Freedom (1992), Blasphemy in Modern Britain 1789-present
(1999) and senior editor for the Journal for the Critical Study of Religion. In
addition to his historical expertise, he has had to draw upon legal studies,
criminology, literary and film studies, area studies and philosophy for
frameworks to explain blasphemy’s history and how and why the phenomenon
persists. His most recent book, Blasphemy in the Christian World (Oxford
University, Press 2007), surveys the history of the subject throughout the West
and speculates upon its future development. He has given verbal and written
evidence to the House of Lords Select Committee on Religious Offences and has
been in considerable demand by the British media to talk upon the subject
(notably BBC Radio Four, TV Channel Four). He is an Executive Officer of the
Social History Society of Great Britain and a Fellow of the Royal Historical Society. Dr Jasdev Singh Rai Dr Jeremy Shearmur Jeremy Shearmur is Reader in Philosophy, ANU, and lectures in political, moral
and social philosophy. He worked for eight years as assistant to Professor Sir
Karl Popper, and is author of The Political Thought of Karl Popper, and Hayek
and After, both 1996, and was co-editor, with Piers Turner, of Karl Popper’s
After the Open Society, 2008. He has taught at the University of Edinburgh, the
University of Manchester, and at George Mason University, where he was a
Research Associate Professor. He was also Director of Studies of the Centre for
Policy Studies, in London. ix Acknowledgements This collection evolved out a two-day conference called ‘Negotiating the Sacred:
Blasphemy and Sacrilege in the Arts’ held at the Centre for Cross Cultural
Research at The Australian National University in November 2005. We would
like to thank the ANU E Press board and the collection’s reviewers for their
suggestions and feedback which have helped shape and polish the collection. Adam Art Gallery, Éditions Typo and Denise Boucher, kindly gave us for
permission for extensive quotations, and Cathy de Monchaux and the estate of
Ian Breakwell gave permission to reproduce images of their artwork. We would
also like to thank the staff of the Centre for Cross Cultural Research, Dr Kevin
White, and the many volunteers from around the ANU, for their contribution
to the success of the conference. This is the second volume from a series of five conferences and edited
collections on the theme ‘Negotiating the Sacred’. The first conference,
Negotiating the Sacred: Blasphemy and Sacrilege in a Multicultural Society, was
held at the ANU’s Centre for Cross-Cultural Research in 2004, and published as
an edited collection by ANU E Press in 2006. Other conferences in the series
have included ‘Religion, Medicine and the Body’ (ANU, 2006), ‘Tolerance,
Education and the Curriculum’ (ANU, 2007), and ‘Governing the Family’ (Monash
University, 2008). xi Elizabeth Burns Coleman and Maria Suzette Fernandes-Dias Behzti, a play by the British Sikh playwright Gurpreet Kaur Bhatti
became the centre of a major controversy in the United Kingdom in December
2004 when the opening night was disrupted by a riot at the Birmingham
Repertory theatre. In A Short History of Blasphemy, Richard Webster suggested that Western
liberals and artists deploy the rhetoric of a holy war in defence of freedom of
expression just as readily as Muslims do in relation to their defence of Mohamed
and the Koran, and indeed, hold the doctrine of freedom of expression nearly
as sacred.1 Lines in the sand have thus been drawn between those wishing to
protect freedom of expression in the arts, and those who think blasphemy is
wrong, and should be suppressed. What is consistent in the liberal position is
that the ‘opponent’ is a ‘zealot’ or an intolerant moralising bigot seeking to
impose his or her particular version of the good upon us all. In his discussion of the British debates over the place of blasphemy law in
Britain, Clive Unsworth sees a polarisation between those who see blasphemy
law as supporting the role of religion and religious values in society, and liberals
who object to blasphemy laws on the basis that the law’s object is to protect
some members of society from being offended, not merely by exposure to the
blasphemy, but through the knowledge that other people may be viewing the
material and not finding it offensive.2 The three events in Britain in the late twentieth century that re-ignited
debates over blasphemy involved ‘the insertion of elements of sexuality and
sexual deviancy into the religious narratives’.3 These events included the Gay
News case, Satanic Verses, and the refusal to license the video Visions of Ecstasy. In the 1977 Gay News case, Mary Whitehouse successfully invoked British
blasphemy law against the editor of the paper, Denis Lemon, for the publication
of the poem ‘The Love that Dares to Speak its Name’ by James Kirkup. Lemon
was sentenced to nine months’ jail, suspended for 18 months, and Gay News
was fined £1000, with prosecution costs awarded against them. The decision
and the interpretation of the crime were upheld by a majority of the House of
Lords. Elizabeth Burns Coleman and Maria Suzette Fernandes-Dias The sacrosanctity of religious dogmas and beliefs, stringent laws of repression
and codes of moral and ethical propriety have compelled artists to live and create
with occupational hazards like uncertain audience response, and accusations of
deliberate misinterpretation of cultural production looming over their heads. In
extreme cases, the battle between artistic iconoclasm and societal repression has
forced creators to put their life on the line in defence of liberal self-expression. Perhaps we should not write in the past tense. On 2 November 2004, Dutch
film maker Theo van Gogh paid with his life for his supposedly offensive
depiction of Islam in his film, Submission which denounced violence against
women in Islamic societies. In February 2005, the ‘Världskulturmuséet’ (‘Museum
of World Culture’) in Göteborg, Sweden decided to remove the painting Scène
d’Amour by Louzla Darabi that was part of a temporary exhibition about
HIV/AIDS, and depicted a man and a woman having sexual intercourse. The
artist and the curator had received numerous death threats, some with the
postscript ‘learn from the Netherlands’, from Muslims enraged over the Koran
quotations that were featured in a corner of the painting. In September 2005,
Europe suffered the violent outbreak of public outrage from the Muslim world
(several dead, embassies burnt and international tension) when the Danish
newspaper Jyllands-Posten printed 12 cartoons of Mohammed. These are dramatic cases in contemporary culture wars, yet the issue is not
merely about the relationship between Western artists and Islam. Banal use of
religious symbols continues to spark controversy as some of us believe that
excessively liberalised and interpretative use of religious symbols/icons/figures
in art, divests these representations of faith, of their sanctity, and, at times,
offends conventional piety. 1998 saw an unsuccessful private prosecution in
New Zealand after the Te Papa museum displayed Virgin in a Condom. Protesters
besieged the museum, and attacked the sculpture. In 2000, the Sensation
exhibition at the Museum of Modern Art in New York was picketed because of
its inclusion of a painting by Chris Ofili, The Holy Virgin Mary, which
incorporated carefully placed elephant dung. The Australian National Gallery
cancelled its display of the exhibition. In 2004, Oliver Stone’s movie, Alexander,
based on the life of the fourth century BC Macedonian king, evoked protests
from the Zoroastrian diaspora for the use of a Zoroastrian holy symbol, the 1 Negotiating the Sacred II Farohar. Elizabeth Burns Coleman and Maria Suzette Fernandes-Dias Unsworth acknowledges that ‘it was not the case that the blasphemous
quality of Kirkup’s poem was dependent upon its homosexual content, and
indeed, supporters of the prosecution were anxious to maintain that the trial
was not about homosexuality but about the vilification of Christ’. Yet, for
Unsworth, the stated objection does not undermine their obvious bigotry: ‘in
the context of the wider belief system, the homosexual content of the poem
cannot have done other than to aggravate the blasphemy and render it exorbitant
in the eyes of those who so judged it’.4 This case started a vigorous campaign by artists and liberals to have the
blasphemy laws abolished, a campaign that resurfaced almost 12 years later in 2 2 Introduction Introduction the wake of the Rushdie affair. The Satanic Verses, a bitter satire on Islam,
understandably gave serious offence in depicting Muhammad as ‘Mahound’, a
calculating opportunist and debauched sensualist, and giving the names of
Muhammad’s wives to prostitutes in a brothel.5 The last case, Visions of Ecstasy, a 1989 short film directed by Nigel Wingrove,
was refused certification by the British Board of Film Classification (BBFC) because
of scenes featuring a sexualised representation of Saint Teresa of Ávila caressing
the body of Jesus on the cross—scenes which could potentially make the film
liable to prosecution for blasphemy. As cutting the scenes would remove
approximately half of the film’s content, the board decided to refuse certification
altogether. In 1996, the distributor of Visions of Ecstasy took his case to the
European Court of Human Rights where the BBFC’s decision to reject certification
was upheld. The Court concluded that: ‘Freedom of expression constitutes one
of the essential foundations of a democratic society. As paragraph 2 of Article
10 expressly recognises, however, the exercise of that freedom carries with it
duties and responsibilities. Amongst them, in the context of religious beliefs,
may legitimately be included a duty to avoid as far as possible an expression
that is, in regard to objects of veneration, gratuitously offensive to others and
profanatory.’6 It is this that liberals are unable to accept. One thing we may agree about is that the protection of religious sentiments
appears to be the legally accepted purpose of blasphemy law. Elizabeth Burns Coleman and Maria Suzette Fernandes-Dias Unsworth writes: The crime of blasphemy is directed to the objective of protecting religious
believers from outrage to their feelings from relevantly offensive material
being in circulation, in the interests of a social value of maintaining
respect for a sense of reverence of the sacred, so that it has to do with
the social status of religion. It is the tenuous and intangible nature of
the harm against which the offence protects that causes especial outrage
to liberal exponents of the pre-eminent value of freedom of expression.7 The crime of blasphemy is directed to the objective of protecting religious
believers from outrage to their feelings from relevantly offensive material
being in circulation, in the interests of a social value of maintaining
respect for a sense of reverence of the sacred, so that it has to do with
the social status of religion. It is the tenuous and intangible nature of
the harm against which the offence protects that causes especial outrage
to liberal exponents of the pre-eminent value of freedom of expression.7 We might be wary of Unsworth’s portrayal of the issue in terms of the nature
of the offence as a kind of prurient attitude towards sexuality. It may be the
case that Mary Whitehouse was seeking to uphold moral sexual values. Yet, in
A Brief History of Blasphemy, Webster reminds us that both Christian polemicists
and Western orientalists for centuries ‘sought to denigrate Islam by attributing
to it a fantastic, disreputable or demonic sexuality’.8 According to Webster,
the Muslim objection to the use of sex within the Satanic Verses was not to sex
itself, but to the use of obscenity as a form of vilification. On the picture Unsworth presents, the conservative position on blasphemy
is to uphold moral (sexual) values in society, and the liberal position is the
standard position against legal moralism, that ‘offence’ is not a justified basis
for the state to interfere with freedom of expression. 3 Negotiating the Sacred II According to Caslon Analytics, an Australian research consultancy, voices
decrying blasphemy against art have been inspired by ‘overseas models, with
local provocateurs, zealots and “concerned citizens” emulating excitement in
London, New York or other cultural centres’.9 The consultancy suggests that
the protests have seldom sustained media attention or support of major religious
groups or community leaders and have encountered a largely indifferent
audience. Elizabeth Burns Coleman and Maria Suzette Fernandes-Dias This characterisation belittles those protesting against blasphemy in
Australia as fringe groups, whose position is not worth considering, but it is
true that, within Australia, the accusation of blasphemy does not immediately
give rise to widespread public ire. Despite the very public claim made by Justice
Harper, Supreme Court of Victoria, when refusing to grant an injunction to
prevent the opening of Andres Serano’s Piss Christ at the National Gallery of
Victoria (Pell v Council of Trustees of the National Gallery of Victoria) that as a
multicultural and tolerant society, Australia ‘need not bother with blasphemous
libel’, the work was vandalised on two different occasions in two weeks,
compelling the gallery to close the exhibition to prevent further assaults against
its staff. The technicalities that quashed the injunction (‘a civil court will not
exercise criminal jurisdiction and will not restrain what may or may not be a
legal act by using a civil remedy such as an injunction’10 ) and defences of the
freedom of expression were not sufficiently potent to quell the passions that the
image aroused in some members of the public and, in others, perceptions of an
infringement of decency and good taste. Webster suggests that blasphemy laws may have come to be considered
obsolete because ‘respect for the figure of Jesus and for Christianity in general
has been inculcated so widely, even among non-believers, that the restraints of
good taste have generally made the restraints of law all but redundant’.11 He
argues that it would be considered ‘an unpardonable breach of good taste’ for
a sceptic or a non-believer to blaspheme in front of a believer, and that this
amounts to an internalisation of the sacredness of Christian religion.12 It is not
the case, however, that the same respect would automatically be extended to
other religious groups within our communities. He speculates that it is because
of such internalised repression that ‘the role of artists, poets, novelists and
film-makers as ‘agents’ of blasphemy has become so important in the twentieth
century’.13 Our aim in this book is to move beyond these portrayals of debates over
blasphemy as a contest between fundamentalism or legal moralism and liberal
freedoms and to re-examine the nature of the offence. To cut through the
entrenched positions about blasphemy and freedom of expression, it is necessary
to recognise that freedom of expression is a political right. Elizabeth Burns Coleman and Maria Suzette Fernandes-Dias The protection of this
right does not mean that all acts of blasphemy are morally permissible or
acceptable. Legal permissibility and moral acceptability are different, and one Introduction may have a political liberty to do something that is morally wrong. Moreover,
the condemnation of repressive acts of violence in response to blasphemy is not
incompatible with the condemnation of gratuitous disrespect of religious symbols
and offence to others. There is a debate to be had, of course, about whether
offence should be taken seriously morally, and whether offence is the correct
characterisation of the wrong involved. The perspective of the religious persons
may differ from that of those who do not share their belief. For legal philosopher
Joel Feinberg’s ‘profound offence’, the ‘wrong’ involved is not, or not merely,
that someone has been offended; it is that they are offended because the act is
wrong.14 In such cases, mere knowledge of the act is sufficient to cause offence. The reason why religious beliefs are particularly susceptible to this form of
offence is that they contain a sacral element that is missing from other strongly
held beliefs.15 (It does not follow from this that religious beliefs are the only
form of belief that might be considered important in this regard, as the issue
concerns the centrality of a belief to a person’s life.) Focusing on ‘fundamentalism’, or ‘zealots’ or ‘legal moralism’ blinds us to
the broader social context. This debate is not merely about the role of Christianity
in our society, but the place of religion in a multicultural society with numerous
religious groups. Issues surrounding the rights of minority cultures, in particular
indigenous cultures, to recognition and respect have raised new questions about
the contemporariness of the construct of blasphemy and sacrilege. Controversies
over the aesthetic representation of the sacred, the exhibition of the sacred as
art, and the public display of sacrilegious or blasphemous works, have ignited
heated debates and have invited us to reflect on binaries like ‘artistic and religious
sensibilities’, ‘tolerance and philistinism’, ‘the sacred and the profane’,
‘deification and vilification’, and to reconsider what actually amounts to
blasphemy and sacrilege in the present day context of multicultural
cosmopolitanism and political secularism. Elizabeth Burns Coleman and Maria Suzette Fernandes-Dias Although blasphemy is traditionally defined as ‘a contemptuous or profane
act, utterance, or writing concerning God or a sacred entity’, ‘the act of claiming
for oneself the attributes and rights of God’ or ‘an irreverent or impious act,
attitude, or utterance in regard to something considered inviolable or sacrosanct’,
the construct has extended itself to the domain of the secular, to include concepts
like the desecration or flagrant disrespect of ‘civil religion’ and mythologies of
nationalism and identity. Sacrilege is conventionally defined as the transgression
against the virtue of religion in terms of ‘violation of a sacred place’, ‘irreverent
treatment of sacred things’, ‘defilement of honour of a “sacred” person’, and it
is not always clear where the line between blasphemy and sacrilege lies. While the writers in this collection have approached the question of
blasphemy and sacrilege very differently, the usages reflect many of the issues
in negotiating the sacred in the arts, and, indeed, highlight how varied these 5 5 Negotiating the Sacred II issues may be. In our first chapter, Nash observes that in ancient and early
Christian periods, blasphemy involved physical attacks on religious artefacts
with the intent to damage the religious ‘currency’ of the religion so attacked. In contrast, medieval and early modern accounts of blasphemers see them as
wilful heretics indulging their own pride, an ‘active commission whereby doubts
or errant opinions were actively vocalised’. David Manning, like Nash, sees
blasphemy as primarily a crime within the Christian tradition, and contrasts the
contemporary interpretation of blasphemy as an offence of people’s sensibilities
with an older interpretation of blasphemy as an offence to God. Coleman develops
a broad interpretation of blasphemy as a failure to display the homage appropriate
to what is represented by an image or symbol in order to extend the concept
into cross-cultural settings, and to understand what is objectionable about
aesthetically appreciating some indigenous religious objects. McGowan, in
contrast, understands the placement of sacred objects in a gallery context not
as blasphemy, but as a form of re-sacralisation. Both Coleman’s and McGowan’s
cross-cultural interpretations would be rejected by Rai, who argues that Western
academics fail to appreciate that there is no clear distinction between the sacred
and the profane in Indic thought. As editors, we have not attempted to impose
a concept of blasphemy or sacrilege; in contemporary debates such words count
as markers of dispute over meanings, contexts, and uses. Elizabeth Burns Coleman and Maria Suzette Fernandes-Dias Our purpose is to
highlight and explore these different uses, and to try to find a way to understand
and negotiate differences between values. The sacred may be presented as an
absolute, but may also be negotiated. Negotiation may mean discussion leading
to some kind of agreement or settlement, but it may also mean to clear or pass
an obstacle. The justifications for artistic freedom in relation to blasphemy include the
claim that art can, or should, be considered an autonomous practice that should
be judged in terms of aesthetic merit; that the artists’ intention, for instance to
make a political comment or to break down barriers, justifies blasphemy, and
that complete freedom of expression is necessary for the creation of art. The first
argument involves the idea that literary genres, galleries, theatres and other
forms of secular, public ‘space’ provide a context that allows us to recognise
that what is occurring is ‘a representation’, and enable a specifically aesthetic
form of interpretation. Many artists assume that such ‘spaces’ establish the
framework for an interpretation and appreciation that is autonomous, and distinct
from moral and religious evaluation. As we grapple with the task of articulating how artists strive against
ideological and religious sanctions to affirm the ‘dissenting voice’ and affirm its
ability to ‘enable people to question and assess the validity of dominant social
norms and their institutionalisation’,16 we are also conscious of the means by
which some achieve this without raising the ire of religious authorities, and
alienating others within our societies. 6 Introduction Introduction The book is divided into four sections. The first provides four different
accounts of blasphemy and sacrilege in the arts and in relation to religious
artefacts that we may find displayed as arts. The second section is concerned
with the motivations that artists may have in deploying blasphemous images. The third looks at two very different interpretations of freedom of expression,
and the fourth is concerned with how artists express themselves from within
religious traditions and despite restrictions. In an age where politicians still use the oldest gimmick of justifying their
oppressive acts as answers to a divine calling and align their motives to religiously
sanctioned agendas, art continues to provide the transgressive space for
subverting dominant ideological discourses. Elizabeth Burns Coleman and Maria Suzette Fernandes-Dias However, despite their emancipatory
power, artists and their art occupy a liminal space wherein the contemporary
socio-political climate of hegemonically-induced extremism and increased
communal and religious sensitivity and intolerance, limit individual freedom of
expression. Artistic articulation of individual conviction without the intention
to offend can still potentially cause unrest. In a world that is becoming
increasingly pluralistic and multicultural, it is necessary to step beyond the
simplistic assertion that free speech should override religious sensitivities and
to facilitate a discourse that will encourage a negotiation of definitions of
blasphemy or sacrilege and a sensitisation of religious sensibilities, and limit the
abusive deployment of freedom of expression. Artistic sophistication and layering
of meaning with its ‘virtues’ of ambiguity, openness, and indeterminacy that
prevent a single, blatant and overt interpretation of a creation as sacrilegious or
blasphemous, presents one such means of reconciling potential conflict. Enabling
and creating an environment of tolerance that is conducive to intellectual debate
and not offence, is yet another. Respect for the religious and cultural sentiments
of others is of the utmost importance, for, it is only in a society that respects
difference that a forum for negotiation can evolve and the lines in the sand can
fade. 1 Richard Webster, 1990, A Brief History of Blasphemy: Liberalism, Censorship and ‘The Satanic Verses’,
Southwold: The Orwell Press.
2 Clive Unsworth, 1995, ‘Blasphemy, Cultural Divergence and Legal Relativism’, Modern Law Review,
vol. 58, pp. 658-677, at p. 665.
3 Ibid
664 8 Webster, op. cit. , p. 40. 13 Ibid.
14 Joel Feinberg, 1985, The Moral Limits of the Criminal Law: Volume 2, Offense to Others, New York:
Oxford University Press, p. 67.
15 Peter Jones, 1990, ‘Respecting Beliefs and Rebuking Rusdie’, British Journal of Political Science, vol.
20, no. 4, pp. 415-437, at p. 424-5.
16 H. Marcuse, 1978, The Aesthetic Dimension, Boston: Beacon Press, p. 9. 15 Peter Jones, 1990, ‘Respecting Beliefs and Rebuking Rusdie’, British Journal of Political Science, vol.
20, no. 4, pp. 415-437, at p. 424-5. 16 H. Marcuse, 1978, The Aesthetic Dimension, Boston: Beacon Press, p. 9. 14 Joel Feinberg, 1985, The Moral Limits of the Criminal Law: Volume 2, Offense to Others, New York:
Oxford University Press, p. 67.
15 15 Peter Jones, 1990, ‘Respecting Beliefs and Rebuking Rusdie’, British Journal of Political Science, v
20 no 4 pp 415 437 at p 424 5 Ibid.
14 Joel Feinberg, 1985, The Moral Limits of the Criminal Law: Volume 2, Offense to Others, New York:
Oxford University Press p 67 Section I The first four essays in this collection express a sense of bewilderment about
accusations of blasphemy and sacrilege, and seek to understand the motivations
behind them. In our first essay, historian David Nash suggests that in
contemporary Western societies, blasphemy ‘is back with a vengeance’ and
‘offers to trail blaze a path for religion and religious responses to become the
cornerstone of the new politics of cultural identity’. However, he argues, we
have no adequate theoretical models for understanding the place of religion in
our societies. The rise of accusations of blasphemy, he suggests, is proof of the
falsity of the secularisation thesis that modernity would bring about a lessening
of religious feeling and greater tolerance. Claims of blasphemy seem to be
increasing in Western societies. Assumptions that the process of liberalisation
and secularisation would lead to the disappearance or privatisation of religion,
and the disappearance of blasphemy as a crime, have been misplaced. Accordingly, secularisation theory does not provide a theoretical framework
through which we can understand the changes occurring in our society. Rather,
he suggests, that blasphemy is more than a transgressive activity. It is a place,
he writes, ‘where identities were forged and communities debated issues about
public order’. David Manning explores the evolution of the charge of blasphemy in England. During the reign of the last two Stuart monarchs, the English society was a
deeply religious society and vexed by a perceived rise in immorality and
irreligion. To the pious majority, licentious behaviour was at best a scourge on
the respectability of society and at worst a threat to the very salvation of the
nation. In 1698, Jeremy Collier’s A short view of the immorality and profaneness
of the English stage, sparked a debate in which the most zealous tracts and sermons
demonised the theatres and accused some plays and actors of blasphemy. The
investigation touches upon the nexus between theology and morality, the
perceived complicity of audiences, and the conception of malicious intention
and interpretation, to present a view of blasphemy that was more than a
one-dimensional notion of intolerability. According to Manning, would-be
reformers such as Tillotson, Collier, and Bedford perceived the nature of the
wrong as the language used in theatre: it was not good or edifying. Endnotes
1 3 Ibid., p. 664. 3 Ibid., p. 664. 4 Ibid., p. 671. 5 5 Shabbir Akhtar, ‘Be Careful With Muhammad!’, cited in Webster, 1990, p. 39. 7 Freedom, the Individual and the Law (1993), p. 254, cited in Unsworth, 1995, p. 667. 8 9 http://www.caslon.com.au/blasphemyprofile4.htm#brian
10 10 http://www.artslaw.com.au/Publications/Articles/97Blasphemy.asp
11 7 Negotiating the Sacred II 12 Ibid. 20, no. 4, pp. 415-437, at p. 424-5. 8 8 Section I The context
of performance, within a play on a stage, which might be considered by
contemporary audiences as the means by which we distinguish between art and
life, did not change the nature of the language. Manning concludes that belief
in a providential God, coupled with conventional theology, may render seemingly 9 Negotiating the Sacred II innocuous plays and facile passages blasphemous and as a tangible threat to the
nation. Order and purity are themes that arise in our third chapter. Elizabeth Burns
Coleman explores the concept of blasphemy in terms of a metaphysical
relationship between the symbol and what it represents, and as a form of
cognitive response to an image that places us in a certain relationship to it. These
theories suggest that very different assumptions are made about the nature of a
symbol to those generally found in the arts. Coleman discusses a seemingly
innocuous poster for the 1998 Adelaide Arts Festival depicting a Madonna with
a piano accordion and the objections that many indigenous peoples have to the
display of their sacred objects within a gallery context. Coleman argues that
these innocuous cases, which display no intentional or explicit attack against
religion, are similar in terms of the lack of honour or respect shown to a symbol. From this, she argues that the concept of honour or respect is central to
developing a cross-cultural concept of blasphemy. She argues that our
comprehension of the meaning of symbols is shown by our actions in response
to them. Our physical responses to symbols are a form of labelling, or
understanding, that show us to be in a certain kind of relationship with them. Concerns about the proper use of sacred images, and about the contexts in which
they appear, are concerns that we show appropriate respect through our
behaviour. Former curator and gallery director, Dianne McGowan tries to encompass
the shifting nature of the concept of sacredness in a secular market economy
oriented society; what was once sacred is now housed in art museums or
decorating private homes or reproduced or appropriated. She analyses two
paintings, Da Vinci’s The Last Supper and a Tibetan Buddhist thangka, titled
Buddha Sakyamuni, painted between 1050-1100. Both paintings were conceived
for religious spaces and depict religious scenes. However, the thangka, on being
consecrated by a Lama, ceased to be an object and became a living deity. Section I The
Last Supper on the other hand was not consecrated, but its presence in a holy
precinct verifies its sacredness. McGowan argues that the gallery context does
not desecrate, but creates a sacred space for viewing religious and sacred art. 10 10 Freedom of expression and its development The reaction to alien views in contemporary worlds and the historic past has
quite often been termed blasphemy. In some respects it is easy to say that the
outpourings of others deserve consideration and recognition whether or not
their discourses or statements are coherent. But history reminds us that we must
confront the consequences of the freedom that the modern world has given us. While those who fought for the extension of forms of democracy in the post-war
world would celebrate the downfall of spurious and enslaving authority, the
gap that this loss of authority left was large and threatening. Indeed, we should
remember that there are important reasons for describing this situation as a gap. This became plain in a significant range of cultural discourses that talked about
the ‘end’ of such reassuring concepts as civilisation, history, and, of course, the
modernist project. Such discourses looked back at supposed species of tyranny
and constraint that had apparently been transcended but scarcely, if at all, looked
forward to a world of potential liberation and open ended expression. For the
optimistic, and those who never gave a second thought to the fact that they
themselves were empowered by culture, the future represented a wealth of
opportunities. Clearly, empowerment was precisely the key to this so-called
liberation. Those who had benefited from the material consequences of the
modernist project, for example, through education, or through access to the
public sphere and the media had simultaneously the wish, the means, and the
ability to express themselves and to influence others. Paradoxically, the
breakdown of confidence in these same mechanisms left others without such
forms of liberation or expression. Moreover, this also contributed to their
long-term loss of confidence in their own empowerment through the
opportunities offered by the West. The breakdown of this confidence emphasises how far we have grown deeply
accustomed to writing an especially Whig, socio-democratic progressivist style
history of both religion and rights in the West. A pejorative attitude to religion
was also often present in modernisation theory which considered superstition
and forms of belief to belong to primitive epochs and that these represented a
phase through which human development passed.1 Traditionally, Whig religious
history has told us that religious tolerance has developed in Western nation
states as a consequence of these states conceding species of human rights. 1
Blasphemy and sacrilege: A challenge
to secularisation and theories of
the modern? David Nash In the penultimate episode of the recent beloved BBC science fiction series Dr
Who the massed ranks of the Daleks made a long awaited re-appearance. In doing
so they demonstrated a developed conception of amoral violence and the
justification for it in a conception of blasphemy. When confronted by the
possibility that they might contain human DNA, the Daleks considered this to
be a suggestion that was potentially blasphemous. This is arguably a quite
significant cultural moment for the confidence that underpinned our modernist
vision of civilisation. The Daleks after all were created in the early 1960s,
arguably the highpoint of post-war confidence in humanity’s capacity to sort
out social and economic problems. At this time, the Dalek was literally a
modernist dustbin into which all that was unsavoury in human interaction was
consigned. Yet the Daleks have returned to a world that has (since they’ve been
gone) seen the concepts of ‘ethnic cleansing’, ‘collateral damage’, ‘hate crime’
and ‘The War on Terror’ emerge. The violent and unsavoury material consigned
to the bin is climbing out to recolonise a world once thought confidently civilised. Moreover, it is concepts like blasphemy and other species of violent attack upon
identity that potentially provides for the emergence of a more confrontational
and violent world. Discourses of blasphemy that might lead to its detection and indeed committal
are back with a vengeance and offer to trail blaze a path for religion and religious
responses. They are also seeking to become the highway leading towards the
new politics of cultural identity. Moreover, discourses of blasphemy are invading
culture itself to become something of a catch-all term that allows individuals to
display frustration and to place distance and difference between themselves and
others. Most of us in the West have considered religious and cultural tolerance
to be an intrinsic and cherished part of the modernist dream. The modern world’s
communications technology seems to present an inescapable power that should
transcend hatred, anger and misunderstanding. These days we can share our
innermost thoughts and feelings with other people without facing or interacting
with them. The internet has made this possible, fostering the growth of our own 11 Negotiating the Sacred II personal world of interests and adding apparent authority and validity to ever
divergent opinions. Today we have an information society that is potentially
overloaded with both facts and opinion. 1
Blasphemy and sacrilege: A challenge
to secularisation and theories of
the modern? But, strangely, this ‘opinion rich’ society
has become a society that has never seen individuals and groups more in conflict. Freedom of expression and its development This
concession of human rights often went hand in hand with these states
surrendering stewardship and authority over the ideological makeup of their
citizen’s moral and mental lives. Generally speaking, this stewardship was 12 Blasphemy and sacrilege: A challenge to secularisation and theories of the modern? considered to be of a religio-moral nature and its withering away within this
historical model of progress was often linked to the sociological theory of
secularisation. In one branch of this theory, offered substantially in the 1960s
by the then influential work of Peter Berger, the supposed authority of religion
in the areas of behavioural morality, education and welfare were deemed to have
been broken by developments within the post-war world.2 Such developments
had arguably been at work since the Enlightenment. Teleological notions of
society’s development from Hegel, Marx, Weber, Elias and Foucault envisaged
the marginalisation of religion into various components. Whether this was
brought about by revolution (Hegel/Marx), greater specialisation (Weber), civility
(Elias), or the growth of the subjectively empowered self (Foucault), all theories
predicted its potential demise. A brief reflection over the last paragraph will alert readers to how old
fashioned the secularisation paradigm sketched actually looks — yet systematic
discrediting of its assumptions and conclusions is still required. Interestingly,
the modernisation and peaceful transition model of religious change has a
stunning longevity and some commentators have noted the capacity for such
accounts to overwrite conflict models.3 In his examination of the archaeological
evidence of religious violence from the classical world, Eberhard Saur examined
how the archaeological processes of excavation and restoration are capable of
denying the phenomenon of iconoclasm. In discussing the public display and
portrayal of one statue (the statue of Mithras from Ostia), Saur notes that its
archaeological value is too often considered through its carefully restored state. In this, the mutilation of the statue has been forgotten and similar evidence of
destruction, so Saur argues, has been neglected or ignored. A factor that obscures
the history of religious conflict still further is what Saur describes as a trend
within archaeological explanations of change to adopt gradualist models of
‘acculturation’. Acculturation models argue for a much more gradual interleaving
of cultures based around peaceful contact, mutual sharing of technology and
cultural achievements — all opposed to ideas of conquest and cultural
imperialism. Interestingly, this bears a striking resemblance to modern
perceptions of the functioning and purpose of religious/multicultural tolerance. Freedom of expression and its development Of still greater concern is Saur’s suggestion that the advocates of ‘acculturation’
theses of change are apt to describe destructive conflict models of change as old
fashioned and shaped by discredited nineteenth century paradigms.4 Similarly, the Reformation was regularly described as a linear unstoppable
process which came fundamentally to alter the social and religious makeup of
the West. In this we are persuaded that the building blocks of modernism
(constructed by More and Erasmus) were firmly in place before Marx, Hegel,
Weber, and even Nietzsche put pen to paper. Likewise, the triumph of the
Newtonian rational universe noted in the history of science and even within the
histories of gambling and probability portrays a modernisation motif which 13 Negotiating the Sacred II explains rationality as the triumphant explanatory framework around which
the universe turns.5 No longer did the turn of the dice or the shape of natural
phenomena or laws display the divine will but instead could be explained
through natural laws or the secular notion of probability. Saur reminds us how archeological models that display destruction and
iconcoclasm are worth taking with us in any study of secularisation as a
consensual linear and modernising history. Attacks upon religious images
represent a significant portion of blasphemy’s early history. Blasphemy in the
ancient and early Christian periods most frequently comes down to the historian
as the written record of individual acts of outrage. Like many other crimes, there
must also have been a dark figure of iconoclastic acts against religious material
culture that have subsequently left no trace. The wilful and calculated destruction
of religious artefacts or buildings displayed an intent to damage the religious
‘currency’ of the religion so attacked. Evidence from the ancient world strongly
suggests that monotheistic religions took the lead in iconoclastic practices and
the identification of other religions as anathema. Pre-Christian Rome had been
indulgent to both empire-wide and local deities and archaeologists have seen
the widespread existence of some cults as tangible evidence of this. Individual
deities were chosen for their usefulness and augmented existing belief systems
rather than displaced them. It was monotheistic religions that set themselves
against this equilibrium.6 The creation of a single autonomous religious truth and route to salvation
provides the obvious explanation as to why this was clearly the case. Blasphemy and the framing of secularisation One relic of modernisation and Whig history that still has lingered longer than
most is the theory of secularisation, although the secularisation theory which
strove to have a dominant place in the history and sociology of religion has now
retreated somewhat in the face of a number of intellectual and theoretical
challenges, particularly from historians. If the theory of secularisiation no longer
claims to offer an overarching explanation of the last millennium of Christianity’s
history, it can at least point to its description of general trends in belief and
outlook as maintaining its validity. Secularisation theory, in various forms,
suggests that the modern world is more secular than the pre-modern, and this
in turn more secular than the medieval. It points to a whole host of indicators
that have altered mankind’s perception of the world and how it functions. These
include the arrival of scientific beliefs to replace belief in providence, the transfer
of education, welfare and medicine into the secular sphere and, lastly, the end
of mass religious attendance accompanied by the retreat of religion from public
life.8 Moreover, secularisation’s theoretical account of the historical past suggests
that episodes such as the Reformation, the rise of Protestantism and the
Enlightenment constitute milestones towards the creation of this secular society. The high watermark of confidence in the theory of secularisation occurred a
generation ago, but it remains arguable that modern society is overall more
secular than previous epochs. However, blasphemy’s history significantly contests the certainty and
inevitability of this view of historical progress and offers a significantly different
chronology for religious history. Certainly, it may be possible to argue that
blasphemy itself represents a ‘secularisation’ of the concept of heresy. Heretics
were quite regularly cast as victims of their errors that entered their minds
unwittingly. Most medieval and early modern accounts of blasphemers see them
as wilful individuals indulging their own pride. The investigations of early
modern Germany, France and Switzerland would lend credibility to this view.9
The offence of blasphemy became one of active commission whereby doubts or
errant opinions were actively vocalised within the earshot of others. In contrast
to heresy, blasphemy was increasingly considered to be a lapse of discipline
rather than sincere religious error. Many medieval European cases display
individuals as drunk or angry at some slight providence had wrought upon
them. Freedom of expression and its development When
Pharaoh Akhenaten adopted monotheism in ancient Egypt the destruction of
rival deities was a logical and inevitable consequence of this choice. Judaism
similarly had scriptural precedent for violent action against pagan deities,
although some of these examples were aimed at religious usurpation or the
Israelites ‘straying’ from the true God. After Christianity’s arrival, the trend was
continued further by Islam’s adoption of iconoclastic practices. As Islam spread
East its iconoclasm left its mark upon the Buddhist shrines and idols of India. Later Christianity also regarded pagan relics and practices to be capable of
routinely contaminating the lives of true believers. The systematic nature of
Christian iconoclasm can be contrasted with the haphazard and inconclusive
actions of Roman predecessors. Sauer suggests that the Emperor Aurelian’s
capture of Palmyra resulted in half-hearted and piecemeal destruction of local
sacred places and, perhaps, even represented a tolerance or respect for the deities
of the defeated city.7 In sum, the linear models of secularisation producing and being sustained
by tolerance and rationality from the Enlightenment to the dawn of modernisation
deserve to be qualified if not wholly questioned. Models of acculturation, of
which secularisation is surely one, can be shown to have overshadowed models 14 Blasphemy and sacrilege: A challenge to secularisation and theories of the modern? of conflict and episodic reaction. Similarly the post-Enlightenment ‘invention’
of religious tolerance ignores the relative tolerance that existed within some
early civilisations. Blasphemy and the framing of secularisation Others attacked the honour of God in anger against the ill fortune he had
bestowed upon them, frequently at the gambling table.10 However, blasphemy’s history significantly contests the certainty and
inevitability of this view of historical progress and offers a significantly different
chronology for religious history. Certainly, it may be possible to argue that
blasphemy itself represents a ‘secularisation’ of the concept of heresy. Heretics
were quite regularly cast as victims of their errors that entered their minds
unwittingly. Most medieval and early modern accounts of blasphemers see them
as wilful individuals indulging their own pride. The investigations of early
modern Germany, France and Switzerland would lend credibility to this view.9 The offence of blasphemy became one of active commission whereby doubts or
errant opinions were actively vocalised within the earshot of others. In contrast
to heresy, blasphemy was increasingly considered to be a lapse of discipline
rather than sincere religious error. Many medieval European cases display
individuals as drunk or angry at some slight providence had wrought upon
them. Others attacked the honour of God in anger against the ill fortune he had
bestowed upon them, frequently at the gambling table.10 Nonetheless, other assumptions of the secularisation account of religion’s
supposedly waning importance do not fit with blasphemy’s very different 15 Negotiating the Sacred II history. The Reformation, far from diluting the sacred, shifted the focus to
scripture and the Bible. New light is also shed on this through noting how
mediaeval conceptions of sin and shame equally had their later Protestant
counterparts.11 The Enlightenment, it might be argued, brought philosophical
and later artistic questioning of the sacred and its place in society. This would
fit with the history of the offence in England where French Revolution ideals
were later replaced by arguments about free speech, individualism, and the
removal of religion to the private sphere. Nonetheless, such a triumph was not
linear, nor was it remotely complete since the link between church and state in
England was never overthrown. Secularisation theory itself has also never been comfortable in measuring
Christianity’s vibrancy within the realm of the private sphere. While it prefers
to see privatisation as the story’s conclusion, recent history has shown the sacred
as capable of being mobilised from this area with a rapidity that would astonish
the previous generation. Blasphemy and the framing of secularisation In Britain at the end of the twentieth century and into
the new millennium, isolationist agendas have sought to protect both Britain
and its component nations from the European super state arguing for ‘opt outs’
in issues around obscenity and morality. In 1996, for example, a case reached
the European Court in which the film maker Nigel Wingrove appealed against
the refusal of the British Board of Film Classification to award an exhibition
certificate on the grounds it breached his freedom of expression. The court
eventually found in favour of the British government allowing it the margin of
appreciation, a means by which important cultural areas that might be covered
by overarching European Union Law would be allowed to opt out from this. Britain thus was allowed to keep a working legal concept of blasphemy alive. More damning evidence still comes from America, where an institutionalised
cleavage between Church and State did not remove the concept of blasphemy
from this society. Nor did the Constitution actively secularise American society,
forcing religion into the private sphere. Religion has become a central part of
American politics and has developed a rhetoric which increasingly attempts to
define the state as secular rather than neutral, and is therefore able to describe
secular stances and world views as unconstitutional if practiced by the
government.12 Even where it might be claimed that religion was successfully
privatised, the conclusions we might draw from the evidence do not support
components of the secularisation thesis. Religion becoming a matter for the
private conscience occurred at precisely the same time as knowledge of the law
and access to it became a real possibility for more individuals in the West. Secularisation theory itself has also never been comfortable in measuring
Christianity’s vibrancy within the realm of the private sphere. While it prefers
to see privatisation as the story’s conclusion, recent history has shown the sacred
as capable of being mobilised from this area with a rapidity that would astonish
the previous generation. In Britain at the end of the twentieth century and into
the new millennium, isolationist agendas have sought to protect both Britain
and its component nations from the European super state arguing for ‘opt outs’
in issues around obscenity and morality. Blasphemy and the framing of secularisation In 1996, for example, a case reached
the European Court in which the film maker Nigel Wingrove appealed against
the refusal of the British Board of Film Classification to award an exhibition
certificate on the grounds it breached his freedom of expression. The court
eventually found in favour of the British government allowing it the margin of
appreciation, a means by which important cultural areas that might be covered
by overarching European Union Law would be allowed to opt out from this. Secularisation theorists argued that tolerance would be bred by greater contact
between individuals, and that globalisation would homogenise religious outlooks
and lessen differences between faiths and groups. In some respects the ecumenical
agendas of Western churches in the 1960s and 1970s represents a good example
of this, as does the modernisation agenda of Vatican II. Similarly, the arrival of 16 Blasphemy and sacrilege: A challenge to secularisation and theories of the modern? scientific beliefs to replace beliefs in providence, and the aforementioned
secularisation of education, welfare and medicine, accompanied the retreat of
religion from public life.13 This confidence has not been borne out by the
evidence of the late twentieth century that has seen individuals reach for
blasphemy as a tool capable of preventing the erosion of the sacred. For some
individuals and groups, defending religion has rejuvenated nationalism in the
face of the twin agendas of globalisation and, in particular, pan-European systems
of morals and justice. In England, blasphemy cases as far apart as the 1880s, the 1930s, the 1970s
and the 1990s, all contained a fear of continental influences contaminating
‘sacred’ English morals. The case against Nigel Wingrove’s film Visions of Ecstasy
allowed the United Kingdom to maintain the Common Law of blasphemous libel
as an element of its culturally distinct identity, similarly Mary Whitehouse had
invoked the sacred nature of British morality in her campaign to exclude Jens
Jurgen Thorsen’s Sex Life of Christ from being shot in England in 1977. In
accepting these as legal and policy decisions, English fears of the dangerous and
foreign were enshrined in and influenced legal decisions.14 Elements of these
fears were evident in the cases against G.W. Foote in 1883, where the relevant
newspaper’s blasphemy could be shown to be directly influenced by French
prototypes. Blasphemy and the framing of secularisation In the 1930s Britain also witnessed an attempt to extend the
blasphemy law as a means of excluding ‘foreign’ Jewish freethinkers and their
ability to undermine the morals of the British Empire. Although this Bill passed
a first reading in Parliament, the government moved swiftly to prevent it
becoming law. The early 1990s saw the Salman Rushdie affair persuade some to
see blasphemy as a multicultural issue. One of the arguments used was the
suggestion that Britain’s tolerance of other religions and cultures was a sham
unless the law could be mobilised to protect the religious sensibilities of all. Early twentieth century America also witnessed a number of panics associated
with the dangerous spirituality and anti-religious views of recent immigrants. In the immediate First World War period, a Lithuanian, Michael Mockus, stood
trial for blasphemy and, upon conviction, the judge made clear reference to the
potentially dangerous views held by immigrants. Mockus linked Christianity
with class and economic oppression and argued that religion was essentially a
conspiracy against the workers of the world.15 Seventeenth century America
had seen individual states operate their own penal codes, but latter day America
also witnessed the use of blasphemy as a tool for the defence of local communities
and their morals. The early twentieth century in particular saw American States
pursuing blasphemers within their local and state jurisdictions (recognisably
similar to early modern Geneva, Zurich or Venice) only to see these attempts to
prosecute and convict quashed by federal law.16 Nonetheless, individual
communities have for some years now been able to operate their own local
standards of decency in defiance of Federal law’s long standing desire for equality 17 Negotiating the Sacred II of practice.17 Thus, secularisation theory clearly does not offer any valuable
framework to explain blasphemy’s history and in particular its persistence. Moreover, it is possible to go further and argue that blasphemy’s own history
itself provided important material for a wider history of the sacred. Its persistence
and its current relevance supply vital evidence of the survival of the sacred —
not simply as a spiritual legacy but as a vital component of beliefs about
nationalism and wider religious identity. Many of our models (especially theoretical ones) concerning long term
continuity and change offer inadequate explanations of blasphemy. The
phenomenon itself does not readily fit with theories associated with
modernisation and change. Blasphemy and the framing of secularisation Following such models and their cultural intentions
leads us to ask why blasphemy was not eradicated through the Enlightenment’s
search for freedom of belief and expression. If theories about the inevitable
demise of religion were true, it could be expected that the crime of blasphemy
would have been swept away by the eighteenth and nineteenth century
revolutions in America and Europe, or fallen into desuetude in the more organic
constitutions of Britain and its former English-speaking empire. In the
non-English speaking world, the imposition of supposedly enlightened laws
from the mother country ironed out local differences to provide what some have
seen as an enduring solution.18 The survival of blasphemy as a crime, and as
an accusation, also offers a temptation to suggest that blasphemy as a cultural
phenomenon may suit a more dystopian view of the West in crisis. Those Western
critics who were (and are) suspicious of the Enlightenment, and its supposed
empowerment of the individual, are the important figures here. Foucault, Derrida
and the postmodernists not only regard this empowerment to be a sham but also
are likely see the supposed authority of Enlightenment truth as a collection of
controls whose ultimate explanatory cohesion disintegrates before them. Blasphemy, identity and exploration Blasphemy was, and is, more than a transgressionary activity. Its long history
suggests that controversies about blasphemy are places where definitions of the
universe and its working are debated; places where identities are forged and
where communities debate issues about public order. All these lead us to a
history away from the certainties offered by secularisation narratives. Blasphemy
always reminded us that the relationship between individuals and the sacred
were very often problematic. Secularisation theorists always spoke about ideas
of collective, normal, and everyday belief, thereby homogenising both the
experience of belief and the definitions that historians and sociologists would
give to it. Blasphemy’s illumination of conflict models and incidents showed
that belief was capable of ebbing and flowing and appearing at pressure points
in the interactions of individuals and societies. In this, the inevitable triumph
of the secular over the sacred looks a far-fetched suggestion. Even if secularisation 18 Blasphemy and sacrilege: A challenge to secularisation and theories of the modern? theorists did have some justification in arguing that religion would become less
important, it remains a teleological theory of progress and continual
enlightenment. But the religion supposedly supplanted by reason was not by
any means destroyed or damaged beyond repair, and secularisation theorists
failed to pay much attention to the residues and legacies that Christianity left
in the West. Artists who explore and contemplate the West’s myths, stories and cultural
legacy became the people who rescued religion from its appointed status on the
periphery of experience. What motivates artistic consideration of the sacred,
and why artists as diverse as Andres Serrano, Bettina Rheims, Tracey Emin and
Tania Kovats have quarried the sacred for inspiration, are important questions. While these artists themselves may have lucid, exciting and informative answers,
the wider ‘influence’ of the sacred needs to form exploration of the fuller context
in this area.19 In this respect, it is valuable to ask how far this art reflects a
re-discovery or re-orientation around the sacred imagery of the past, or a simple
acknowledgement of religion’s enthralling nature as one of the West’s strongest
and most compelling ‘stories’. In this, it has not been for many of these artists
a re-evangelisation of life, or necessarily a quest for spiritual longing, but has
equally sometimes been spawned by the simple curiosity of those brought up
with secular outlooks. Blasphemy, identity and exploration Films, and their
subsequent greater availability on video, are capable of multiple consumption
and re-consumption in the quest for favourable or unfavourable meanings.20
But the engagement of artists and writers with the religious has provided the
opportunity for religious world views to challenge ownership of these cultural
ideas and have asked the searching question of whether the ideas of the religious
belong to society as a whole. The protests against everything from Last
Temptation of Christ to Jerry Springer the Opera have given religious individuals
the opportunity to bring arguments about the sacred and its inviolability back
to the public sphere. In this they are aided by agendas which promote social
inclusiveness and the urge to marginalise differences of opinion and viewpoints
in the more consensually driven societies of the new millennium. In particular, we might note how images and portrayals of Christ on film
have caused especial problems of interpretation sometimes leading to accusations
of blasphemy. Of greater concern have been the attempts to humanise Christ
which, in some eyes, appeared to be an assault upon the idea of his divinity —
a constant struggle in such depictions. Dennis Potter’s Son of Man portrayed a
Christ at war with himself and sceptical of the divine nature of his own destiny. Similarly, Scorsese’s Last Temptation gave critics, both hostile and sympathetic,
an opportunity to air the sheer variety of interpretation to which the gospel
story, and its depiction, could be subjected. Yet, conclusions from this area were
not readily transferable to other blasphemous instances. Films, and their
subsequent greater availability on video, are capable of multiple consumption
and re-consumption in the quest for favourable or unfavourable meanings.20 But the engagement of artists and writers with the religious has provided the
opportunity for religious world views to challenge ownership of these cultural
ideas and have asked the searching question of whether the ideas of the religious
belong to society as a whole. The protests against everything from Last
Temptation of Christ to Jerry Springer the Opera have given religious individuals
the opportunity to bring arguments about the sacred and its inviolability back
to the public sphere. In this they are aided by agendas which promote social
inclusiveness and the urge to marginalise differences of opinion and viewpoints
in the more consensually driven societies of the new millennium. These are all conundrums and challenges to artists and thinkers. 1 A classic example of this paradigm in use is Keith Thomas’s 1971 Religion and the Decline of Magic,
London: Weidenfeld and Nicholson.
2 Peter Berger, 1967, The Sacred Canopy: The Social Reality of Religion, Garden City, N.Y.: Doubleday.
3 We might contrast the Berger version of secularisation with that offered by Bryan Wilson, in his later
Religion in Sociological Perspective (Oxford: Oxford University Press 1982). This envisaged religion
ceasing to be important in the working of the social system but instead becoming personalised or
privatised. Again we may wish to note how Western and Christocentric this analysis is, even for its
time. Bryan Wilson also suggested that the process could be measured through churches becoming,
among other things, mosques! Blasphemy, identity and exploration This interest means that investigations of popular culture and media studies
will inevitably be drawn to the sporadic appearance of the sacred in popular
culture. In a sense, each particular media can be said to have its fifteen minutes
in the spotlight. Irving Welsh showed the power of disreputable images of the
almighty to shock in the context of the modern novel. His Granton Star Cause
depicted a profane and drunken deity and was the last controversy Mary
Whitehouse, Britain’s premier twentieth-century critic of media morality, was
involved in before her death. Madonna’s sense of melodrama almost inevitably
took her in the direction of the sacred long before she thought of actually being
crucified on stage. In 2006, her ‘Confessions’ tour played with Catholic motifs
and extended her belief that suffering and crucifixes could be ‘made sexy’. Even
culturally conservative British comedy, which always wishes it was more
subversive than it ever manages to be, produced a publication with religious
offensiveness as a keynote feature. The British comedian Rik Mayal’s recent
autobiography contained a picture of himself beatified and described as ‘better
than God’. All these are in danger of demonstrating a de facto cheapening and
ubiquitousness of the sacred while noting the rediscovery of religion’s powers
to shock. Yet equating these with outright blasphemy brings problems of
intention when, for example, Maddona had embarked on a quest to entertain
as much as to problematise or inform her audience about religion and the sacred. 19 Negotiating the Sacred II In particular, we might note how images and portrayals of Christ on film
have caused especial problems of interpretation sometimes leading to accusations
of blasphemy. Of greater concern have been the attempts to humanise Christ
which, in some eyes, appeared to be an assault upon the idea of his divinity —
a constant struggle in such depictions. Dennis Potter’s Son of Man portrayed a
Christ at war with himself and sceptical of the divine nature of his own destiny. Similarly, Scorsese’s Last Temptation gave critics, both hostile and sympathetic,
an opportunity to air the sheer variety of interpretation to which the gospel
story, and its depiction, could be subjected. Yet, conclusions from this area were
not readily transferable to other blasphemous instances. Blasphemy, identity and exploration In such a
climate, it is scarcely surprising that a cultural phenomenon so capable of
controversy should excite and fascinate the artistic mind used to sifting and
redefining the problem areas of civilisation. In the end, we should protect such
rights and imperatives since they offer a blueprint for possible change against
regimes of abject quietism and stasis. This is necessary because all culture —
religious or secular — is capable of development and change. Whether we really
can provide an equality of public space for the religious and those who would
denounce it to re-enter the public sphere is a difficult question for us to answer. Ironically, societies faced by these dilemmas will have to decide how far they
can tolerate a conflict model between the religious and the free speech advocate
in the name of so-called consensus. Endnotes
1 20 Blasphemy and sacrilege: A challenge to secularisation and theories of the modern? 4 Eberhard Sauer, 2003, The Archaeology of Religious Hatred in the Roman and Early Medieval World,
Stroud: Tempus Books, pp. 15-18. p
pp
5 See Gerda Reith, 1999, The Age of Chance: Gambling in Western Culture, London: Routledge. 6 5 See Gerda Reith, 1999, The Age of Chance: Gambling in Western Culture, London: Routledge. 6 See Sauer, op. cit. 6 See Sauer, op. cit. 7 Sauer, ibid., pp. 162-164, 30, 159, 46 and 162. 7 Sauer, ibid., pp. 162-164, 30, 159, 46 and 162. 8 Secularisation theory and its own history can be approached through D.S. Nash, 2004, ‘Secularization’s
failure as a master narrative; Reconnecting Religion with Social and Cultural History’, Cultural and
Social History, vol. 1, no. 2, pp. 302-325. 9 See Gerd Schwerhoff, 1998, ‘Starke Worte: Blasphemie als theatralische Inszenierung von Männlichkeit
an der Wende vom Mittelalter zur Frühen Neuzeit’ in Martin Dinges (ed.), Hausväter, Priest, Kastraten:
Zur Konstruktion von Männlichkeit in Spätmittelalter und Früher Neuzeit, Göttingen, pp. 238-263; Alain 9 See Gerd Schwerhoff, 1998, ‘Starke Worte: Blasphemie als theatralische Inszenierung von Männlichkeit
an der Wende vom Mittelalter zur Frühen Neuzeit’ in Martin Dinges (ed.), Hausväter, Priest, Kastraten:
Zur Konstruktion von Männlichkeit in Spätmittelalter und Früher Neuzeit, Göttingen, pp. 238-263; Alain
Cabantous, 2002, Blasphemy: Impious speech in the West from the seventeenth to the nineteenth century,
Eric Rauth (trans.), New York: Columbia University Press; and Francisca Loetz, 2002, Mit Gott handeln:
von den Zürcher Gotteslästerern der Frühen Neuzeit zu einer Kulturgeschichte des Religiösen, Vandenhoeck
and Ruprecht Göttingen. p
g
pp
Cabantous, 2002, Blasphemy: Impious speech in the West from the seventeenth to the nineteenth century,
Eric Rauth (trans.), New York: Columbia University Press; and Francisca Loetz, 2002, Mit Gott handeln:
von den Zürcher Gotteslästerern der Frühen Neuzeit zu einer Kulturgeschichte des Religiösen, Vandenhoeck
and Ruprecht Göttingen. 10 For examples of this, see Maureen Flynn, 1995, ‘Blasphemy and the Play of Anger in sixteenth
century Spain’, Past and Present, vol. 149, pp. 29-56, p.35-36.; Maureen Flynn, 1998, ‘Taming Anger’s 10 For examples of this, see Maureen Flynn, 1995, ‘Blasphemy and the Play of Anger in sixteenth
century Spain’, Past and Present, vol. 149, pp. 29-56, p.35-36.; Maureen Flynn, 1998, ‘Taming Anger’s
daughters: New treatment for emotional problems in Renaissance Spain,’ Renaissance Quarterly, vol. 51, no. 3, pp. Endnotes
1 864-886, and Javier Villa-Flores, 2007, ‘On Divine Persecution: Blasphemy and Gambling
in New Spain’ in Susan Schroeder and Stafford Poole (eds.), Religion and Society in Colonial Mexico, New
Mexico University Press: New Mexico. 11 It is worth comparing Delumeu’s thesis about late medieval Christendom’s own obsession with guilt
alongside Max Weber’s version for post-Reformation Protestant Europe. See Jean Delumeau, 1990
edition Sin and Fear: The Emergence of a Western Guilt Culture 13th 18th Centuries Eric Nicholson 11 It is worth comparing Delumeu’s thesis about late medieval Christendom’s own obsession with guilt
alongside Max Weber’s version for post-Reformation Protestant Europe. See Jean Delumeau, 1990
edition, Sin and Fear: The Emergence of a Western Guilt Culture, 13th-18th Centuries, Eric Nicholson
(trans.), New York: St. Martin’s Press. 12 Marjorie Heins, 1993, Sex, Sin, and Blasphemy: A guide to America’s censorship wars, New York: The
New Press. 13 Secularisation theory and its own history can be approached through D. S. Nash, 2004, 13 Secularisation theory and its own history can be approached through D. S. Nash, 2004,
‘Secularization’s failure as a master narrative; Reconnecting Religion with Social and Cultural History’,
Cultural and Social History, vol. 1, no. 2, pp. 302-325. ‘Secularization’s failure as a master narrative; Reconnecting Religion with Social and Cultural History’,
Cultural and Social History, vol. 1, no. 2, pp. 302-325. Interights and Article Nineteen 1995, Blasphemy and Film Censorship, submission to the European Court
of Human Rights in respect of Nigel Wingrove v. the United Kingdom. Article 19/Interights, London. 15 Interights and Article Nineteen 1995, Blasphemy and Film Censorship, submission to the European Court
of Human Rights in respect of Nigel Wingrove v. the United Kingdom. Article 19/Interights, London. 15 15 See L. Levy, 1993, Blasphemy, Verbal offense against the sacred, from Moses to Salman Rushdie, New
York: Knopf, pp. 513-515. See also Theodore Schroeder, 1970, [1919], Constitutional Free Speech Defined
and Defended in an Unfinished argument in a case of Blasphemy, New York: De Capo Press, re-print of
1919 edition published by the Free Speech League. 15 See L. Levy, 1993, Blasphemy, Verbal offense against the sacred, from Moses to Salman Rushdie, New
York: Knopf, pp. 513-515. See also Theodore Schroeder, 1970, [1919], Constitutional Free Speech Defined
and Defended in an Unfinished argument in a case of Blasphemy, New York: De Capo Press, re-print of
1919 edition published by the Free Speech League. Endnotes
1 16 Such prosecutions became impossible after Burstyn v Wilson, 343, U.S. 495 (1952). 1 18 See the Indian Criminal Code ‘imposed’ in 1860 which equalised the legal situation between all
religious groups in India by outlawing all attacks upon religious artefacts and buildings. Indian Penal
Code (1860), sections 295, 295a and 298. 18 See the Indian Criminal Code ‘imposed’ in 1860 which equalised the legal situation between all
religious groups in India by outlawing all attacks upon religious artefacts and buildings. Indian Penal
Code (1860), sections 295, 295a and 298. 19 A recent contribution in this area is S. Brent Plate, 2006, Blasphemy: Art that Offends, London: Black
Dog Publishing. 19 A recent contribution in this area is S. Brent Plate, 2006, Blasphemy: Art that Offends, London: Black
Dog Publishing. 20 It is worth noting also that individual films are probably just as likely to be viewed in the wider
canon of their director than as a manifestation of a particular genre. For an example pertinent to this
area, see Lawrence S. Friedman, 1997, The Cinema of Martin Scorsese, Oxford: Roundhouse, and Ian
Christie and David Thompson, 2003, Scorsese on Scorsese, London: Faber. 20 It is worth noting also that individual films are probably just as likely to be viewed in the wider
canon of their director than as a manifestation of a particular genre. For an example pertinent to this
area, see Lawrence S. Friedman, 1997, The Cinema of Martin Scorsese, Oxford: Roundhouse, and Ian
Christie and David Thompson, 2003, Scorsese on Scorsese, London: Faber. 21 2
‘The devil’s centres of operation’:1
English theatre and the charge of
blasphemy, 1698-1708 David Manning The dominant notion of blasphemy in Britain today is built upon a perception
of what is offensive to the Christian religion. Derived from common law and
fashioned by the dynamics of public opinion, this view has been employed to
powerful effect in the Gay News trial of 1977 and, most recently, in charges
against the BBC broadcast of Jerry Springer the Opera in 2005.2 It would appear
that the exclusively religious conception of blasphemy as a sin against God,
punishable by God or His intermediaries, has been displaced by a more secular,
politicised view which focuses on the relationship between human agents. The
paradigm created by a broadly secular western democracy invariably places
artistic representations of the sacred within a polarised debate between religious
sensibilities and freedom of expression. In such a context, the need to negotiate
the sacred becomes imperative to maintaining a modus vivendi. Practically, a
model of upholding individual rights without risking a breach of the peace is
one to be defended. However, if our intellectual discussions of the sacred become
limited by such a framework, then we restrict our understanding of beliefs that
are not similar to our own. It should be clarified that there are other, more
profoundly religious, perceptions of blasphemy, and to conflate them with the
notion dominant in Britain threatens the potential for us to negotiate the sacred
successfully. By using an historical case study, this chapter will explore an
exclusively religious context to blasphemy and, in so doing, reveal what might
cause a devotedly religious individual to make the charge of blasphemy. Here,
the application of history becomes particularly useful, because we can escape
immediate bias that the charge of blasphemy is only made by cranks or fanatics,
as well as preconceptions concerning the right of freedom of expression.3 Let
us, then, consider the reaction of pious Christians to the activities of the theatre,
within the context of a devoutly Christian society. Blasphemous theatres and plays The only legislation ever passed with Royal Assent against blasphemy in England
was the Blasphemy Act of 1698.4 Political wrangling had limited the scope of 23 Negotiating the Sacred II the Act to a denunciation of anti-trinitarian conceptions of God; however, it was
sold to the public as a realisation of King William III’s personal commitment to
‘discourage profaneness and immorality’.5 The necessity of this presentation
was borne out of a belief among growing numbers of Protestants that England
was plagued by immorality and vice on a scale that risked divine punishment. At this time, England was a devoutly Christian society, and a belief in God’s
providence was virtually universal. Indeed, all major events were interpreted
providentially and nationwide fasts were often instigated by the regime to
appease the Almighty in times of perceived crisis. Furthermore, the most pious
individuals believed that God shaped the very nature of human happiness and
misery.6 Those seized by the belief that vice plagued the nation had to look no
further than the earthquake of 1692 for confirmation: God was angry. For zealous
reformers, one of the most public and institutionalised centres of vice was the
playhouse; for it had a ‘natural and unavoidable tendency to that which [was]
sinful and unlawful’.7 Unlike the tavern or the whore-house, the theatre
stimulated the mind as well as the body. It could promote many different vices
to a diverse audience, who shared the experience together in place and time.8
The playhouse was not simply a venue where vice was represented and then
imitated by vulnerable members of the audience;9 it was a place where
playwright, player and audience met to collude in sin. Given that the state had
seemingly failed to provide adequate direction on the matter, it was therefore
timely that the pious clergyman Jeremy Collier published A short view of the
immorality and profaneness of the English stage in 1698.10 Collier’s work proved popular as most upstanding Christians would have
probably acknowledged that some stages, particularly those at the annual
Bartholomew Fair, attracted disreputable characters and promoted vices such
as profane swearing, mocking of religion, lewdness and drunkenness.11 As well
as attacking these vices, however, Collier posited a much more unpalatable
argument that the theatre as an institution was fundamentally evil. Blasphemous theatres and plays Within the
pages of his substantial book he exposed specific plays as immoral, profane, and
blasphemous; furthermore, he lambasted audience appreciation of these plays
as tantamount to worshipping the Devil.12 The ensuing controversy, which has
become known to literary scholars as ‘the stage debate’, questioned the very
existence of the playhouse in a godly society, and served as a searing indictment
of past and contemporary regimes for their failure to define and condemn
blasphemy in all its forms. In 1698, there were two professional, permanent theatres in London, one in
Drury Lane and the other in Lincoln’s Inn Fields, and they were attended by a
wide cross-section of society.13 The Drury Lane theatre was known as the
‘Royal’, being patronised by the monarch, and its actors assumed the title of
‘His Majesties Servants’. The idea that these playhouses could harbour evil was 24 ‘The devil’s centres of operation’: English theatre and the charge of blasphemy, 1698-1708 seen by many contemporaries as preposterous. While there was general support
for Collier’s call to reform the stage, few reformers endorsed his specific charges
of profanity and blasphemy. By 1699 it became clear that parliament would not
sanction any reform of the theatre, and support for the cause gradually waned. Indeed, most contemporaries would have probably forgotten all about Collier’s
charges of blasphemy had it not been for another providential act of God, and
an astonishing historical accident. On the evening of 26 November 1703, a great storm hit England which caused
widespread damage and thousands of deaths.14 The event was universally
acknowledged by the nation’s Protestants as a sign of God’s wrath. To mark the
severity of the situation, Queen Anne declared that a general fast be observed
as public penance on 19 January 1704. Amidst the national distress, it was
reported that a group of actors had only days after the storm performed a version
of Shakespeare’s The Tempest.15 The failure of the playhouse to cancel this
production in the aftermath of the storm caused pious campaigners once again
to take up the pen and attack the stage, this time with a clear mandate from the
Almighty. Blasphemous theatres and plays Collier quickly published a short, cheap summation of A short view
which, along with a number of other zealous anti-stage publications, were bought
and distributed in significant numbers as part of a massive propaganda campaign
by the newly formed Society for the Promotion of Christian Knowledge (SPCK). In the following months, two prominent journalists, John Tutchin and Daniel
Defoe, joined the fray by attacking the stage in their respective publications:
The Observator and A Review. Two years later, the zealous clergyman Arthur
Bedford capitalised on the momentum of the renewed campaign and published
The evil and danger of stage-plays which claimed to list approximately 1400
examples of ‘swearing, cursing and blasphemy’ in plays of the previous two
years alone.16 Despite the ideological differences that would have existed
between them, Collier, Bedford, Tutchin, and Defoe shared a common bond by
stressing the profanity and blasphemy of the theatres, re-igniting the belief that
playhouses were the ‘Devil’s centres of operation’.17 The rest of this chapter
will investigate why these men considered certain plays to be blasphemous. It
will be shown that, far from a one-dimensional notion of offensiveness, the
charge of blasphemy articulated complex and deeply held theological anxieties. A religious critique of dramatic language The interpretations that our anti-stage writers placed on dramatic texts might
well appear facile or even absurd to modern readers and so, before turning to
the evidence, we need to understand the way in which they would have
approached play scripts. In proving specific plays guilty of blasphemy, it would
appear that to men such as Collier, textual and/or dramatic context was a complete
irrelevance. To explain why intelligent men could neglect the fundamental
significance of context, most literary scholars have concurred with Aubrey 25 Negotiating the Sacred II Williams that the clash between Collier and the playwrights was principally the
result of opposing philosophical theories concerning the separation between life
and art. This explanation cast Collier as a staunch defender of the Platonic notion
that representation is a copy of reality, while placing the playwrights in an
opposing Aristotelian camp which stressed that representation is merely a symbol
of mental states that has no direct connection with reality. A number of
playwrights did construct replies to Collier from an Aristotelian perspective;18
however, I would argue that this evidence has guided scholars to make certain
assumptions about Collier’s position without considering the fact that he was a
devout clergyman. Indeed, it would be almost inconceivable that Collier’s method
was not religiously inspired. Before A short view was published, John Tillotson, the then Archbishop of
Canterbury, delivered a sermon against the stage based upon Ephesians 4:29:
‘Let no corrupt communication proceed out of your mouth, but that which is
good to the use of edifying, that it may minister grace unto the hearers’.19 In
preaching that ‘corrupt communication was evidence of a corrupt and impure
heart’,20 Tillotson affirmed the decree of scripture to be absolute and he
seemingly drew upon Augustinian theology to connect evil deeds to an evil
will.21 The implications for the stage were clear: in discerning between good
and evil acts, context was an irrelevance. The significance of this judgment was
fully acknowledged by Collier in the pages of A short view and in similar fashion
Bedford stated that, ‘whatever is a sin when spoken in another place is as much
a sin when spoken in the play-house’.22 Thus, any word or words could be
legitimately censured, no matter what the context, if the accuser could
convincingly argue that they were wicked. The perceived attack upon God’s providence Shortly after the storm, Collier wrote that during the notorious production of
The Tempest the ‘audience were pleased to clap at an unusual length of pleasure
and approbation’ at the mention of chimneys being blown down.25 The accuracy
of this claim is suspect,26 yet it demonstrates how the providential judgement
of the storm had increased religious sensitivity to matters of timing and position
vis-à-vis both action and reaction. In this instance, the subject matter of the play
was broadly irrelevant, for it was completely superseded by the failure of the
playhouse, the actors and the audience to act appropriately in the wake of God’s
divine judgement. The production of the play in question would most likely
have been unrecognisable to audiences today. The play was probably one of
two radical adaptations of Shakespeare’s original work, which included
significant alterations to the script and a good dose of devilish foolery and
bawdyness.27 Nevertheless, it would appear that it was the mere fact that a
‘mock tempest’28 had been played soon after the real, divinely sanctioned
tempest that caused charges of profanity and blasphemy. Indeed, when the day
of national penance arrived, the Bishop of Oxford told the Lords gathered at
Westminster Abbey that the playing of The Tempest shortly after the storm was
an ‘unprecedented piece of Profaneness’ that was an ‘affront to God, unparalleled
by any civilized nation’.29 The reaction to the playing of The Tempest was
certainly exceptional but for our anti-stage writers it was just one example among
many that demonstrated not just a disregard for God’s providence, but a
fundamental corruption of it. As Collier put it, the players and playwrights had
‘attempted as it were to scale the sky, and attack the seat of omnipotence: they
have blasphemed the attributes of God, ridiculed his providence’.30 To ignore
the providential acts of the Almighty was represented by anti-stage critics as a
terrible attack upon God, mocking His powers as if He did not exist. In Sir John Vanbrugh’s play, The Relapse, the character Young Fashion
schemes to seduce his brother’s fiancée in order to secure her substantial dowry. A religious critique of dramatic language It seems evident, therefore, that it
was an acute religious sensitivity to the word of God, rather than crude literalism
or Platonic theory that formed the central component of our anti-stage writers’
concerns. To appreciate their position fully, all intellectual preconceptions
regarding the legitimacy of context and the separation between life and art need
to be set aside. With this interpretative framework established we can now
consider what caused our anti-stage writers to charge plays with blasphemy. Both Collier and Bedford were often explicit about identifying certain words
in play scripts as blasphemous; but the words were also understood within the
wider category of profanity. For example, in A short view, Collier devoted a
chapter to ‘the profaneness of the stage’, which was subdivided into two sections:
‘cursing and swearing’ and ‘abuse of religion’. The former was acknowledged
as being the most prevalent, while the latter, though less common, was perceived
as the more dangerous, principally because sometimes it did not ‘stop short of
blasphemy’.23 Profane swearing, such as ‘e Gad’ or ‘Lard’,24 was quickly
covered to give prominence to offences that were deemed blasphemous. It should,
therefore, be stressed that swearing against God or any form of general 26 ‘The devil’s centres of operation’: English theatre and the charge of blasphemy, 1698-1708 irreverence towards God or Christianity, while contemptible and profane, was
not actually viewed as blasphemy. Upon reading the anti-stage writings of the
period, it can be shown that evidence used to prove plays guilty of blasphemy
clustered around two central themes: corrupting the idea of God’s providence
and the use of demonic language. The perceived attack upon God’s providence Upon devising a workable ploy with his accomplice, Fashion declares,
‘providence thou see’st at last, takes care of men of merit’.31 Collier was so
incensed by these words that he singled them out as ‘plain blasphemy’ and ‘an
eruption of hell with a witness’.32 The charge of blasphemy was acknowledged
and taken seriously, for it was one that playwrights did much to discredit. In
defending his work, Vanbrugh stated: ‘every body knows the word providence
in common discourse goes for fortune and yet no one ever thought it blasphemy 27 Negotiating the Sacred II to say, fortune’s blind, or fortune favours fools’.33 Michael Cordner has
suggested that Collier’s allegation of blasphemy was predominantly levelled in
reaction to a playwright’s failure to construct pious sentences from a common
religious vocabulary.34 This interpretation of literary literalism seems rather
one-dimensional. It should be noted that acts of providence were believed to be
instances of God’s omnipresent governance, which were designed to protect the
good and give warning and punishment to the wicked.35 The slightest suggestion
that God could smile upon wicked men would have been seen by Collier as
impossible, and not only an affront to the providential workings of God, but a
sign of devilish pride. In another example, Collier denounced the following words from Thomas
D’Urfey’s version of Don Quixote as ‘directly blaspheming the Creation, and a
Satyr upon God Almighty’:36 Providence that form’d the Fair Providence that form’d the Fair In such a charm Skin, Their Outside made his only Care, And never look’d within.3 In this passage, the last two lines communicate the blasphemy because they were
seen as giving rise to the suggestion that God’s providence was only concerned
with the outward and superficial. The idea that God did not care for a person’s
soul would clearly render much of the Christian faith meaningless. The seemingly
innocuous passage from Don Quixote, to the hyper-sensitive reader, was
tantamount to an open denial of the truth of God’s working in the world. It
would appear that the charge of blasphemy allowed Collier to bypass lengthy
explanation and communicate the true wickedness of the passage to less astute
persons or those believing the words to be harmless. Invoking devils Many anti-stage writers expressed a belief that the playhouse was the Devil’s
domain. The origin of this belief dated back to the writings of the influential
second-century Christian Tertullian, who reported on an instance where a woman
attended a Roman theatre and became possessed by a devil.42 I have yet to come
across any reports of early eighteenth-century audiences being overcome by
such forces and even actors who directly invoked the Devil were not explicitly
described as being possessed, though their words were often denounced as
blasphemous. Indeed, the evidence used to censure plays as blasphemous appears
to have relied heavily on quotations that explicitly referred to devils and
damnation. Such language was not uncommon in plays of the period, being used
metaphorically and more readily as alternative modes of expression to taking
God’s name in vain. The plays in question, however, rarely contained
anti-religious themes, and most contemporaries would have deemed the language
fairly innocuous once set in context. Nevertheless, phrases which, for example,
appeared to represent acts of swearing by ‘death and the Devil’43 were plucked
out of scripts by men such as Bedford and denounced with vitriol as instances
of ‘unparalleled blasphemy’.44 For anti-stage critics, to mention demons in
swearing, cursing, or just an exclamation was evidence of actually calling upon
the forces of darkness. Before turning to the evidence, it would be useful to
establish the conceptual relationship between words that invoked devils, the
charge of blasphemy, and the notion that the theatre was demonic. In The evil and danger of stage-plays, Bedford stated in no uncertain terms
that blasphemy was ‘a sin of that heinous nature, which the damned in Hell are
guilty of, and which makes them incapable of recovery’.45 At first glance, this
belief seems to be forged from an inaccurate reading of Matthew 12:31: ‘All
manner of sin and blasphemy shall be forgiven unto me: but the blasphemy
against the Holy Ghost shall not be forgiven unto men’,46 whereby Bedford
gives no consideration to the scriptural differentiation between pardonable and
unpardonable blasphemy. It transpires, however, that Bedford’s perception of
blasphemy shows remarkable similarities to the doctrine put forward by St. The perceived attack upon God’s providence Blasphemy was a highly
evocative term; contemporary readers would have been well aware of God’s
terrible punishment of blasphemies related in the Bible.38 While it seemed
unbelievable that such a passage could be wicked, the fact that a clergyman
labelled them as blasphemous would have meant that they could not be assumed
harmless with a clear conscience. In his newspaper, The Observator, Tutchin attacked an adaptation of
Shakespeare’s Macbeth which had also been performed shortly after the Storm.39
Contrary to the reaction against The Tempest, Tutchin’s grievance lay squarely
on the content of the play. He believed that the occult, evil forces prevalent in
the text of the play were presented as indistinguishable from God’s providential
powers, perverting the unique and infinitely good character of God.40 As it has
been shown, the fictional status of the characters was no excuse; real people
were still saying the words in the script and those words existed in print. God
alone was providential and so, according to Tutchin, the play was proof that
the actors were ‘impious and blasphemous wretches’ who had ‘ridiculed God’s 28 ‘The devil’s centres of operation’: English theatre and the charge of blasphemy, 1698-1708 Amazing and Stupendous Judgment’ of the Great Storm. The severity with
which Tutchin viewed this transgression against God’s providence was made
clear when he concluded that, ‘this is a sad omen of our [the nation’s] hasty
destruction’.41 It is, therefore, evident that the charge of blasphemy
communicated a very grave sin indeed. Invoking devils Thomas Aquinas: to speak any sort blasphemy was a mortal sin which rendered
the utterer damned and those that were damned also blasphemed, to show their
hatred of the punishments inflicted upon them.47 Invoking devils was, therefore,
an example of the latter case (for who but the damned could possibly utter such
words). Put plainly, it was believed that only the damned could call upon devils 29 Negotiating the Sacred II and such an act was blasphemous because it demonstrated that the sinner, even
in the grip of God’s divinely sanctioned punishment, continued to defy the
Almighty. Collier similarly concluded that the profanity of the stage was ‘the
language of the damned’48 and that to pay to watch such sin was to ‘make a
contribution for blasphemy, and raise a tax for the government below?’49 In
demonological theory, the damned were understood to be controlled by devils.50
Therefore, actors who revealed themselves to be damned by their blasphemies,
could be construed as the devil’s agents and consequently to attend the theatre
was, as Tutchin put it, to ‘buy vice at the Devil’s shop’.51 In A short view, Collier attacked John Dryden’s version of Amphitryon 52 for
furnishing Jupiter with the omnipotence of God. To Collier, this was ‘blasphemy
on the top of the letter, without any trouble of inference, or construction’.53
The representation of heathen gods on stage could have been seen as idolatrous
and a violation of God’s first commandment. Yet, in stating that the ‘cover of
an idol is too thin a pretence to screen the blasphemy’,54 Collier hinted that the
sin of blasphemy was of an even greater magnitude. In this case, it would appear
that the notion of blasphemy went well beyond blurring the mythos of the ancient
gods with the ethos of the one true (Christian) God. Early Christian Fathers such
as Tertullian had claimed that Roman gods were in fact devils and for men such
as Collier, who were knowledgeable of and believed in such early Christian
writings, this view continued to hold credibility.55 Implicit in Collier’s treatment,
Jupiter was neither simply a representation of a mythical figure nor a potential
religious idol, but a devil. Invoking devils In light of Collier’s belief in the conceptual relationship
between blasphemy and the demonic, it would appear that the charge of
blasphemy articulated a belief that Dryden had sought to award omnipotent
powers to a devil, thus subverting the differentiation between good and evil
and laying waste to God’s truth. In giving examples of the ‘blasphemous’ expressions of the stage, Tutchin
turned to the words of Vanbrugh’s Provok’d Wife: ‘Satan and his equipage;
woman tempted me, lust weakened me, and so the Devil overcame me, as fell
Adam, so fell I’.56 It would appear that the words that evoked the charge of
blasphemy were ‘the Devil overcame me’, as they could have been construed as
an open admission of a compact with the Devil. Such a view is confirmed when
Tutchin later went on to censure the phrase ‘hail powers beneath!’ from John
Corny’s version of Metamorphosis.57 Bedford denounced similar passages from
other plays that displayed a relationship between human and devil, such as the
invocation ‘to the Devil, so she bring no children’, from the anonymous play
The roving husband reclaim’d.58 On occasion, Bedford perused this interpretation
with a zeal that seems positively ludicrous to the modern reader. For example,
he condemned the phrase as ‘a devil a wit’ from the play The northern lass, which
he annotated ‘i.e. no wit’.59 For Bedford, these words constituted praise to the 30 ‘The devil’s centres of operation’: English theatre and the charge of blasphemy, 1698-1708 devil, which he believed was the ‘highest blasphemy that mortals can invent’.60
It seems clear that the meticulousness and fervour of Bedford’s interpretation
was but a genuine reflection of his fear of God’s wrath and an attempt to
implement what he believed to be God’s wishes as directed by Scripture, earlier
Christian writings and above all else God’s unmitigated providential warning. It has been shown that our anti-stage writers genuinely believed that the
blasphemies of the theatre denigrated the truth of God’s workings in the world
and advanced the position of devils. In both method and substance, this view
was supported by a combination of Scripture, early Christian writings and the
work of eminent Christian theologians. Significantly, Tutchin, Bedford and
Collier were not considered cranks and their views held significant credibility
in the period of heightened theological anxiety after the storm of 1703. Invoking devils To these
men, the godly reformation required to stave off further divine punishment
simply could not be completed unless the theatre was drastically reformed. In
the final section of this chapter I want to return to the issue of negotiating the
sacred and reflect upon how our anti-stage writers sought to apply their examples
of blasphemy to affect actual reform. The plight of God-fearing Christians First of all, it must be stressed that, for pious English Protestants, the notion of
the sacred extended well beyond a collection of discrete beliefs, words, actions
or objects.61 In general terms the sanctity of God’s truth was universally accepted
and so playwrights, audiences and anti-stage writers all worked within, and
agreed upon, the same religious paradigm. This chapter has shown that the act
of negotiating the sacred in the stage debate was not complicated by questions
concerning the nature of the sacred or how it should be accommodated, but
rather how sensitively providence should be interpreted and how vigorously
God’s truth should be maintained and protected. It has already been noted that
our anti-stage campaigners sought more forceful and comprehensive reform than
did the governing regime, and yet God’s providential intervention of 1703 made
it impossible for the administration wholly to dismiss the claims of our writers. Consequently, it would appear that in a state that held the sanctity of God’s
truth as its governing paradigm, the correct interpretation of God’s will rested
exclusively on His perceived intermediaries (i.e. the Monarch and her Bishops). Yet, in times of crisis, the paradigm also allowed for a circumvention of the
authority of the Church and the law because God was ultimately the arbiter.62 The main strategy employed by reformers against immorality and lower-level
profanity was to appeal to magistrates to be more effective in the punishment
of the wicked.63 Yet, it is notable that this approach was not adopted by Collier,
Tutchin and Bedford. The main reason for this was probably pragmatic: it would
have been futile to appeal for state intervention against words that the state did 31 Negotiating the Sacred II not recognise as sinful.64 However, I would suggest that the unique way in
which our anti-stage writers conceived of the theatre as the devil’s domain also
significantly influenced their approach. Their consistent reference to the theatre
as the ‘Devil’s chapel’, or the ‘Devil’s engine’, or the ‘Devil’s shop’65 took the
literal observations of the early Christian Fathers and re-interpreted them afresh
for the eighteenth century by playing upon contemporary fears about the
prevalence of evil and socio-religious concerns among reformers that the theatre
had become anti-church. Importantly, the labels of theatre as the Devil’s ‘chapel’,
‘engine’ or ‘shop’ also indicate how our anti-stage writers believed that the sins
of the theatre were perpetuated. The plight of God-fearing Christians When the writings of our anti-stage writers are
taken in the round, it becomes clear that the main target of the campaign was
not the sinful playwrights and actors, but rather potential theatre goers and
people of influence. It was believed that the effectual reform of the playhouse
lay with the source of the problem — the audience. Collier’s view that the
audience were complicit has already been noted above, and Defoe was under no
illusion that the ‘Errors of the Stage, lie all in the Auditory’ for the actors were
but their ‘Humble Servants’.66 Indeed, it was the audience’s attendance, money
and applause that supplied the Devil with his congregation and ensured that
God’s truth continued to be denied. The outrage created by such evil
‘competition’ and the extent to which the charge of blasphemy exposed
fundamental and dangerous untruths is evident in a satirical poem by Daniel
Defoe, which was published on occasion of a new theatre being built at the
Haymarket in 1705. An extract of the poem reads as follows: View but our Stately Pile, the columns stand,
Like some Great Council Chamber of the Land:
When Strangers view the Beauty and the State,
As they pass by, they ask what Church is that? Thinking a Nation, so Devout as we,
Ne’r build such Domes, but to some Deity;
But when the Salt Assembly once they View,
What Gods they Worship, how Blaspheme the True;
How Vice’s Champions, Uncontrol’d within,
Roul in the very Excrements of Sin:
The Horrid Emblems so Exact appear,
That Hell’s an Ass, to what’s Transacted here.67 That Hell’s an Ass, to what’s Transacted here.67 The primary approach of our anti-stage writers was, therefore, to appeal to the
Christian conscience of the potential theatre goer, thus bypassing the worldly
restrictions of politics and law and asking each individual to scrutinise their
actions more closely. The polemical claims of Collier, Tutchin and Bedford formed a desperate
attempt to expose the wickedness of the stage. Citing play passages as 32 ‘The devil’s centres of operation’: English theatre and the charge of blasphemy, 1698-1708 blasphemous was crucial to the argument that the words in particular were
indeed fundamentally evil. The plight of God-fearing Christians Inflammatory passages attacking God’s providence
and invoking Devils were of little significance in isolation; but if they were
proved blasphemous, then anti-stage writers hoped that audiences would be
forced to accept that plays were evil and risked provoking God’s wrath. With
no earthly, objective judge, such issues were intractable and, for the victimised
playwrights and no doubt many humble audience members, it was actually men
such as Collier who were guilty of blasphemy by seeing wickedness where there
was none. As one eminent playwright put it: ‘where the expression is
unblameable in its own clear and genuine signification, he [Collier] enters into
himself like the evil spirit, he possesses the innocent phrase, and makes it bellow
forth his own blasphemies’.68 After His intervention in 1703, God did not
venture to give any further clarification on the matter and with His apparent
acquiescence, the heat of the stage debate subsided. The SPCK propaganda
campaign wound down in 1708 drawing to a close any serious chance of theatre
reform on the basis of its profanity and blasphemy. Endnotes pp
7 John Edwards, 1705, The preacher discourse shewing what are the particular offices and employments
of those of that character in the Church, (part 1), London. p. 100. 8 8 I would suggest that fundamental similarities between the mechanisms employed by the theatre and
church provide a significant sub-plot to the whole controversy which has yet to be acknowledged by
scholars. For an introduction to some of the main themes from another period, see Bryan Crockett, 1995,
The Play of Paradox: Stage and Sermon in Renaissance England , Philadelphia, PA: University of
Pennsylvania Press. 9 This view is implicit in another contemporary argument that the theatre glorified vice and downplayed
virtue. See John Dunton, The Athenian Mercury, vol. 6, no. 17 (22 March 1692); vol. 8, no. 25 (22 Nov. 1692); vol. 12, no. 7 (14 Nov. 1693). 10 For an excellent and seemingly ignored study of Collier’s life and work see E.A. Ewan, 1961, ‘A
Study of the Works of Jeremy Collier’ (D.Phil thesis, Oxford University). A short view went through
four editions in the first eighteen months and evoked about twenty replies in the same period, see g
y
p
p
Robert D. Hume, 1999, ‘Jeremy Collier and the Future of the London Theatre in 1698’, Studies in Philology,
vol. 96, no. 4, pp. 480-511, p. 495. 11 Sybil Rosenfeld, 1960, The theatre of the London fairs in the 18th century, Cambridge: Cambridge
University Press. 12 12 Jeremy Collier, A short view (London, 1698), unpaginated preface and contents pages. 13 13 Richard Burridge, A scourge for the play-houses: or the character of the English-stage (London, 1702);
Harold Love, 1980, ‘Who were the Restoration Audience?’, The Yearbook of English Studies, vol. 10, no. 1, pp. 21-44. 14 Daniel Defoe, The storm: or a collection of the most remarkable casualties and disasters which happen’d
in the late dreadful tempest both by sea and land (London, 1704). 15 The Tempest was played on 1 December at the Lincoln’s Inn Fields theatre, see: Emmett L. Avery
(ed.) 1960, The London Stage 1660-1800, vol. 2, Carbondale, IL: Southern Illinois Press, pp. 49-50. 16 Arthur Bedford, The evil and danger of stage-plays (Bristol, 1706), p. 81. 17 17 Curtis and Speck, ‘The Societies for the Reformation of Manners’, p. 49. Acknowledging modern providentialism In conclusion, it has been shown that for pious English Christians of but three
centuries ago, the charge of blasphemy was a very grave one indeed. Conceptually, blasphemy was conceived as a form of profanity that ranged from
cursing through to an inversion of God and the Devil. Extreme blasphemies
sought to subvert some of the most fundamental Christian truths, and were
denounced as irredeemable sins. Practically, it has been demonstrated how a
belief in a providential God, coupled with relatively conventional theology,
rendered seemingly innocuous plays and facile passages blasphemous and thus
a tangible threat to the whole nation. This chapter has shown how the meaning of art can be construed from an
exclusively religious point of view. As a result, we should perhaps be more
ready to acknowledge that for those who believe in a providential God and
active devils, philosophical theories of representation and/or political rights of
freedom of expression pale into insignificance. A strong belief in providentialism
is certainly in evidence in many countries throughout the world and we need
only look to the Evangelical Christian lobby in the USA or the Islamic regime
under President Mahmoud Ahmadinejad in Iran to see the vitality and influence
of such beliefs. To assume that all charges of blasphemy against art are limited
to notions of offensiveness between human agents, even within the West,
significantly limits our understanding of how profoundly religious individuals
and societies conceive of the world they live in. To devout believers defending
the sacred is a matter of life and death. 33 Negotiating the Sacred II Endnotes 1 T.C. Curtis and W.A. Speck, 1976, ‘The Societies for the Reformation of Manners: a case study in the
theory and practice of moral reform’, Literature and History, vol. 3, no. 1, pp. 45-64, at p. 49. I would
like to thank my supervisor, Mark Goldie, for his comments and advice throughout the writing of this
paper. I am also grateful for the travel grants given to me by the Arts and Humanities Research Council
and Clare College, University of Cambridge, which enabled me to attend the ‘Blasphemy and Sacrilege
in the Arts’ conference. 1 T.C. Curtis and W.A. Speck, 1976, ‘The Societies for the Reformation of Manners: a case study in the
theory and practice of moral reform’, Literature and History, vol. 3, no. 1, pp. 45-64, at p. 49. I would
like to thank my supervisor, Mark Goldie, for his comments and advice throughout the writing of this
paper. I am also grateful for the travel grants given to me by the Arts and Humanities Research Council
and Clare College, University of Cambridge, which enabled me to attend the ‘Blasphemy and Sacrilege
in the Arts’ conference. 2 For a valuable introduction to the development of this form of blasphemy see: David Nash, 1999,
Blasphemy in Modern Britain 1789 to the Present, Aldershot: Ashgate. 2 For a valuable introduction to the development of this form of blasphemy see: David Nash, 1999,
Blasphemy in Modern Britain 1789 to the Present, Aldershot: Ashgate. 2 For a valuable introduction to the development of this form of blasphemy see: David Nash, 1999,
Blasphemy in Modern Britain 1789 to the Present, Aldershot: Ashgate. 3 I would suggest that the Salman Rushdie affair, for example, has introduced significant challenges
to a constructive debate of blasphemy. 4 4 9 & 10 Guill. III. c. 32. 4 9 & 10 Guill. III. c. 32. 5 Journal of the House of Lords, vol. XVI, p. 175: opening of the ninth session of Parliament, December
3, 1697. 5 Journal of the House of Lords, vol. XVI, p. 175: opening of the ninth session of Parliament, December
3, 1697. 6 John Spurr, ‘Virtue, Religion and Government’: the Anglican Uses of Providence’ in Tim Harris; Paul
Seaward and Mark Goldie (eds) 1990, The Politics of Religion in Restoration England, Oxford: Blackwell,
pp. 29-47. Endnotes 18 18 For example William Congreve, see Aubrey Williams, 1975, ‘No Cloistered Virtue: Or, Playwright
versus Priest in 1698’, Publications of the Modern Language Association, vol. 90, no. 2, pp. 234-246. f
g
g
19 John Tillotson, Fifteen sermons on several subjects, (London, 1702), pp. 289-323. 20 Tillotson, Fifteen Sermons on several subjects, p. 301. 21 For Augustine, ‘an evil will is the efficient cause of an evil deed’, and so it may be suggested that
an evil deed is evidence of an evil will. See Augustine, The City of God, XII, Ch. 6, translated by Philip
Levine (London: Heinemann,1966), p. 25. Levine (London: Heinemann,1966), p. 25. 22 Bedford, The evil and danger of stage plays, p. 188. 23 Collier, A short view, p. 60. 34 ‘The devil’s centres of operation’: English theatre and the charge of blasphemy, 1698-1708 24 The switching of vowels in words such as ‘God’ was a technique used by playwrights to avoid
infringement of the 1605 Act ‘for the preventing and avoiding of the great abuse of the holy name of
God, in stage-plays’ (3 Jac. I c. 21). It is curious to note that a similar technique is still employed today
to circumvent censure, for example in the British television sitcom Father Ted, the word ‘fuck’ is replaced
with ‘feck’ to allow for a pre-watershed broadcast. 24 The switching of vowels in words such as ‘God’ was a technique used by playwrights to avoid
infringement of the 1605 Act ‘for the preventing and avoiding of the great abuse of the holy name of
God, in stage-plays’ (3 Jac. I c. 21). It is curious to note that a similar technique is still employed today
to circumvent censure, for example in the British television sitcom Father Ted, the word ‘fuck’ is replaced
with ‘feck’ to allow for a pre-watershed broadcast. p
25 [Jeremy Collier], Mr. Collier’s dissuasive from the play-house; in a letter to a person of quality,
occasion’d by the late calamity of the tempest (London, 1704), p. 16. For the offending passage in the
play, see Sir William D’Avenant, Macbeth a tragedy with all the alterations, amendments, additions and
new songs (London, 1695), p. 20. 25 [Jeremy Collier], Mr. Collier’s dissuasive from the play-house; in a letter to a person of quality,
occasion’d by the late calamity of the tempest (London, 1704), p. 16. Endnotes For the offending passage in the
play, see Sir William D’Avenant, Macbeth a tragedy with all the alterations, amendments, additions and
new songs (London, 1695), p. 20. g (
)
26 Due to his religious convictions Collier never actually attended a theatre. 27 26 Due to his religious convictions Collier never actually attended a theatre. 27 27 John Dryden and Sir William D’Avenant, The Tempest, or the enchanted Island (London, 1670);
Thomas Duffett, The Mock-Tempest: or the enchanted castle (London, 1675). For further details on the
content and significance of these adaptations see Montague Summers, 1922, Shakespeare adaptations:
the tempest, the mock tempest, and King Lear, London: J. Cape, Introduction; Christine Dymkowski (ed.)
2000, The Tempest, Cambridge: Cambridge University Press; Sandra Clark (ed.) 1997, Shakespeare made
fit, London: Dent, pp. xli-lxxviii; 28 William [Talbot], A Sermon preach’d before the Lords spiritual and Temporal in Parliament assembled,
in the Abbey-Church of Westminster, on Wednesday, Jan. 19. 1703/4. (London, 1704), p. 17. 29 Ibid. 28 William [Talbot], A Sermon preach’d before the Lords spiritual and Temporal in Parliament assembled,
in the Abbey-Church of Westminster, on Wednesday, Jan. 19. 1703/4. (London, 1704), p. 17. 29 b d 30 [Collier], Mr Collier’s dissuasive, op. cit., p. 6. 30 [Collier], Mr Collier’s dissuasive, op. cit., p. 6. 31 John Vanbrugh, The Relapse (1697), p. 19; cited in Collier, Short view, op. cit., p. 84. 32 C lli
it
84 g
32 Collier, op. cit., p. 84. 33 [John Vanbrugh], A short vindication of the relapse and the provok’d wife, from immorality and
prophaneness (London, 1698), p. 25. 33 [John Vanbrugh], A short vindication of the relapse and the provok’d wife, from immorality and
prophaneness (London, 1698), p. 25. 34 Michael Cordner, 2000, ‘Playwright versus priest: profanity and the wit of Restoration comedy’, in
Deborah Payne Fisk (ed.), The Cambridge Companion to English Restoration Theatre, Cambridge: Cambridge
University Press, p. 215. y
(
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University Press, p. 215. 35 William Sherlock, A practical discourse concerning a future judgement (2nd edn. London, 1692), pp. 14-65. 36 35 William Sherlock, A practical discourse concerning a future judgement (2nd edn. London, 1692), pp. 14-65. 36 Collier, op. cit., pp. 196-197. 37 Thomas D’Urfey, The comical history of Don Quixote, (London, 1694), part 1, p. 20; cited in, Collier,
op. cit., pp. 196-197. 38 2 Samuel 12:14 and Ezekiel 35:12-15. 39 38 2 Samuel 12:14 and Ezekiel 35:12-15. 68 William Congreve, Amendments of Mr. Collier’s False and Imperfect Citations (London, 1698), pp.
4-5. Endnotes 39 Sir William D’Avenant, Macbeth, a tragedy. With alterations, additions, and new songs (London, 1674). 40 40 G.R. Elliott, 1958, Dramatic providence in Macbeth: a study of Shakespeare’s tragic theme of humanity
and grace, Princeton, NJ: Princeton University Press. and grace, Princeton, NJ: Princeton University Press g
y
41 Tutchin, The Observator, vol. 2, no. 77 (29 Dec/1 Jan 1704). 41 Tutchin, The Observator, vol. 2, no. 77 (29 Dec/1 Jan 1704). 42 Tertullian, De Spectaculis, XXVI; T.R. Glover (trans.) 1966 , London: Heinemann, p. 291. 43 43 Vanbrugh, The confederacy (1706), p. 29; cited in, Bedford, The evil and danger of stage-plays, p. 40. 44 g
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f
g p y
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44 Bedford, The evil and danger of stage-plays, ibid., p. 40. 44 Bedford, The evil and danger of stage-plays, ibid., p. 40. 45 Ibid., p. 27. 46 45 Ibid., p. 27. 46 Also see, Mark 3:28-30 & Luke 12:10. 47 Aquinas, Summa Theologica 2ae XIII. 48 Collier, A short view , op. cit., p. 96. 49 [Collier], Mr Collier’s dissuasive, op. cit., p. 15. 50 Philip C. Almond, 1994, Heaven and Hell in Enlightenment England, Cambridge: Cambridge University
Press, pp. 86-87. 51 Tutchin, The Observator, vol. 2, no. 31 (21-24 July 1703). 2 52 John Dryden, Amphitryon: or the two Sosia’s (London, 1690). For details of the play and Dryden’s
adaptation in particular, see Örjan Lindberger, 1956, The Transformations of Amphitryon, Stockholm:
Almqvist and Wiksell, pp. 90-120. 52 John Dryden, Amphitryon: or the two Sosia’s (London, 1690). For details of the play and Dryden’s
d
l
Ö
db
h
f
f
h
kh l Almqvist and Wiksell, pp. 90-120. 53 Collier, A short view, op. cit., p. 183. 35 35 Negotiating the Sacred II 55 Tertullian, De Spectaculus, X, cited in Collier, A short view (1698), p. 255. Also see William Bisset
St M
l B
27 M
h 1704
57 55 Tertullian, De Spectaculus, X, cited in Collier, A short view (1698), p. 255. Also see William Bisset at
St. Mary-le-Bow, 27 March 1704, p. 57. St. Mary-le-Bow, 27 March 1704, p. 57. 56 Vanbrugh, The Provok’d wife (1697), p. 77; cited in, Tutchin, The Observator, vol. 2, no. 78 (1-5
January 1704). 57 John Corny, The metamorphosis: or, the old lover out-witted (London, 1704), p. 14; cited in, Tutchin,
The Observator, vol. 4, no. 46 (5-8 September 1705). Endnotes The Observator, vol. 4, no. 46 (5-8 September 1705). 58 [Anon.], Roving husband reclaim’d (London, 1706), p. 8; cited in, Bedford, The evil and danger of
stage-plays, p. 41. 59 Richard Brome, The northern lass (1684), p. 5; cited in, Bedford, The evil and danger of stage-plays,
p. 41. 60 Bedford, The evil and danger of stage-plays, p. 41. 61 Justin Champion, 1992, The pillars of priestcraft shaken: the Church of England and its enemies,
1660 1730 C
b id
C
b id
U i
i
P
6 7 61 Justin Champion, 1992, The pillars of priestcraft shaken: the Church of England and its enemies,
1660-1730, Cambridge: Cambridge University Press, pp. 6-7. 1660-1730, Cambridge: Cambridge University Press, pp. 6-7. 62 I have endeavoured to sketch only the most basic outline of the contemporary debate necessary to
sustain my argument. 63 Tina Beth Isaacs, 1979, ‘Moral crime, moral reform, and the state in early eighteenth century England:
a study of piety and politics’, PhD thesis, University of Rochester, NY. 64 There is no evidence to suggest that the theatres, playwrights or actors suffered legal censure for
blasphemy. However, there were a few instances where actors were prosecuted for lower level profanity
and licentious behaviour. For an authoritative account of these cases, see: Joseph Wood Krutch, 1924,
Comedy and Conscience after the Restoration, New York: Columbia University Press, pp. 167-178. 6 65 For ‘Devil’s chapel’ see: Tutchin, The Observator, vol. 2, no. 77 (December 29-January 1, 1704); for
‘Devil’s Engine’ see, Defoe, A Review, vol. 3, no. 95 (8 Aug. 1706); for ‘Devil’s shop’ see, endnote 53. 66 Daniel Defoe, A Review, vol. 2, no. 26 (May 3, 1705). 67 36 3
Madonna and piano accordion:
Disrupting the order of the world1 Elizabeth Burns Coleman In 1997, a poster depicting an icon of the Madonna playing a piano accordion
was produced to promote the Adelaide Arts Festival. The Madonna was depicted
enthroned against a background that was recognisably of the park that extends
from the Adelaide Arts Centre to the Torrens River, with one of the major
cathedrals in the background. She was surrounded by outlines of gods from
other religions, including Buddha, Ganesh, and an Aboriginal Wandjina. The
style of presentation was recognisably Middle Eastern, a point emphasised by
the gold mosaic frame, and the ‘Greek’ lettering of the announcement of the
event and its main sponsor, Telstra. It appeared, by all publicity standards, to
be a fantastic poster. It was beautiful, and it was rich with connotative association
that made it appropriate for the festival. Adelaide is known as ‘The city of
churches’, so the poster was very specifically appropriate to the Adelaide arts
festival, as opposed to an arts festival in some other place. A third feature of the
poster that made it so good was that the image of the Madonna with an accordion
is inclusive of what may be considered a form of high art as well as popular
culture. Moreover, the image was inclusive of different religious and community
groups and the art of different cultures and, as such, reflected and celebrated a
multicultural recognition of the value of the arts across different cultures. However, within weeks, the poster was withdrawn from circulation by the
Festival organisation because the Greek Orthodox Church had complained it
was an inappropriate and offensive use of one of their icons. Since 1978, the Zuni people of Mexico have been demanding the repatriation
of their war gods, or Ahauutas, from museums around the world, including the
Smithsonian. Their case involves two issues: first, in Zuni eyes the Ahauutas are
communal property and by definition, it is claimed, consider their ownership
by museums not ownership but theft. The second issue is one that concerns us
here. According to James Clifford, ‘Zuni vehemently object to the display of
these figures (terrifying and of great sacred force) as “art”. They are the only
traditional objects singled out for this objection’.2 This same kind of issue may
arise in relation to the display of replicas of sacred objects. 3
Madonna and piano accordion:
Disrupting the order of the world1 In 1975, an ethnologist
at the National Museum of Canada wrote to the director suggesting the removal 37 Negotiating the Sacred II of Iroquoian false face masks on the basis that ‘The Iroquois consider the masks
highly sacred and even “dangerous” objects that should be viewed only at the
time of curing rituals’.3 The masks were removed five years later, but a proposal
to replace them with fibreglass casts of the original was objected to by the Grand
Council of Hodenosaunee (or Iroquois Confederacy). The Council denied the
existence of a distinction between ritually and commercially carved masks and
forbade the exhibition of any masks. It also forbade all forms of reproduction,
and the distribution of any information about them.4 Both the poster and the display of indigenous artefacts appear to be completely
innocuous. The poster did not defile, ridicule or attack Mary or the Church, and
the display of the Zuni war god, and the Iroquois false face masks, was simply
that — their display. So the objections expressed by the Orthodox Church and
the Zuni are mysterious. It is hard to understand why they are objecting, let
alone why we might care about their objections. Both cases challenge important
values we hold. One case confronts the value of freedom of expression. The
second confronts the value of knowledge and the practice of collection for the
purposes of study, public education and aesthetic appreciation. I will argue these two cases are similar, and that this similarity shares features
in common with blasphemy, despite the fact that it is not obvious that either of
these cases are instances of blasphemy. The attempt to understand what is
‘wrong’, that is, to grasp what appears to be mysterious in both these cases, will
provide us with a better understanding of blasphemy. The structure of this
chapter will be as follows. In the first part, I address the idea that these cases
are dissimilar, because they involve a symbol on the one hand, and an object on
the other. I argue that there is no clear distinction between an object and a
symbol. I will also address the concern that the intention to blaspheme, or to
offend, is a necessary element of blasphemy. 3
Madonna and piano accordion:
Disrupting the order of the world1 In the third part of the chapter I
will draw out some of the important features of these icons and beliefs about
their connection to the order of the universe in order to show how we might
understand these cases to be connected to each other, and to blasphemy. Definitions of blasphemy In contrast to the delicacy with which the Adelaide Arts Festival treated Orthodox
sensibilities in relation to the poster including an image of the Virgin Mary, the
National Gallery of Victoria displayed no such misgivings about the display of
Andres Serrano’s controversial photograph, Piss Christ, which showed an image
of a crucifix immersed in the artist’s urine, as part of the Melbourne City Festival
in 1998. This image might be considered paradigmatic as a case of blasphemy. Various Christian churches sought the removal of the image from public display,
and after the negotiations with the gallery and government proved fruitless, the
Catholic Archbishop sought a legal injunction against the gallery displaying the
image, on the grounds that it was blasphemous, indecent and obscene at law. 38 Madonna and piano accordion: Disrupting the order of the world According to Anthony Fisher and Hayden Ramsey, the Archbishop’s action was
supported by the leaders of other Christian churches, as well as the Jewish and
Muslim communities.5 Justice Harper of the Supreme Court of Victoria commented in his refusal of
the application for the injunction that the photograph ‘of itself’ might be seen
as ‘inoffensive’,6 on the grounds that it was beautiful, and showed the crucified
Christ in a golden light; the knowledge that the image was created by immersing
a crucifix in urine is clearly connected to sacrilege. Sacrilege is the violation of
sacred things, and blasphemy, according to Fisher and Ramsey, may be defined
as speaking against God or the sacred or ridiculing things consecrated to God
or held sacred.7 However, as Fisher and Ramsey point out: ‘most sacrileges do
imply blasphemy in the narrower sense of ridiculing the sacred’.8 Yet, it might
be argued, a plastic crucifix is not a sacred object, at least in any legalist
theological sense. A plastic crucifix has not been dedicated or consecrated, or,
as is done with church bells, baptised. But a plastic crucifix is a symbol; it
represents the sacred. Hence, we should not think that there is a clear distinction
between the use of an object, and the use of a symbol. However, these ‘innocuous’ cases do not appear to be cases of blasphemy,
because there does not seem to be any involvement of a deliberate offence. Deliberate offence, that is, intentional offence, is intrinsic to some definitions of
blasphemy. Definitions of blasphemy Fisher and Ramsay reported that, as the scandal progressed, the artist repudiated 39 Negotiating the Sacred II his earlier claim that his art is simply colourful and deliberately shocking, and
asserted that, instead of intending to scandalise, ‘his goal all along had been to
increase the devotion of his fellow Christians by helping them identify better
with Christ in his pain, suffering an humiliation’.10 In contrast to the definition of blasphemy in terms of intentional offence,
Frank J. Hoffman understands blasphemy to be a family of concepts that may
be explicated by determining what the religious points are in each case, and
determining what social practices exist for homage and desecration in each
case.11 Hoffman’s culturally relative account is consistent with Fisher and
Ramsay’s position that objects symbolising mysteries ‘can only be understood
in the context of the religious culture and history of practice from which they
emerge’.12 Hoffman argues, against Perret, that there is no single perspective
from which to define blasphemy. He points out that blasphemy, like obscenity,
is often dependent partly on the attitude of the hearer or recipient. He illustrates
this point with an ‘amusing’ story: Suppose, for example, that there is a pet parrot who is capable of uttering
a perverse litany impugning the religious focal points of the ‘great
religions’ of the world. Now, just as the parrot has completed impugning
the God of Christianity along with the rest, Aunt Millie, being a
frightfully devout (and easily scandalised) Christian lady, indignantly
rushes in from the kitchen with the carving knife, ready to lop off the
poor parrot’s head. Feathers fly, and in the wink of an eye, the badly
mangled parrot has sung his (black) swan-song. Aunt Millie is unlikely
to be repentant at the thought that the parrot did not intend to
blaspheme.13 Despite this unpleasant picture of the people who make claims that something
is blasphemous as the kind of people who would kill parrots, it does seem
arbitrary to insist on a strictly intentional definition of blasphemy, supported
by an appeal to religious authority. Definitions of blasphemy For instance, Roy Perret defines blasphemy as ‘an illocutionary act
which is a function of the agent’s complex intention. In the case of a blasphemous
speech act, the speaker intends that the hearer should come to believe something
through the recognition of the speaker’s intention that the hearer do this’.9
Perret supports this intentional interpretation of blasphemy as theologically
correct with a quotation from Thomas Aquinas’ Summa Theologica: …a man failing to advert to the blasphemous nature of his words, and
this may happen through his being moved suddenly by his passion so
as to break out into words suggested by his imagination, without heeding
to the meaning of the words: this is a venial sin, and is not blasphemy
so called. (Summa Theologica II,ii,13) …a man failing to advert to the blasphemous nature of his words, and
this may happen through his being moved suddenly by his passion so
as to break out into words suggested by his imagination, without heeding
to the meaning of the words: this is a venial sin, and is not blasphemy
so called. (Summa Theologica II,ii,13) This definition suggests that neither kind of case I mentioned, of the display of
the Zuni war gods or of the use of the icon, could be considered cases of
blasphemy. Neither obviously involves the intention to offend, or a propositional
content that the speaker intended the hearer to come to believe. This definition suggests that neither kind of case I mentioned, of the display of
the Zuni war gods or of the use of the icon, could be considered cases of
blasphemy. Neither obviously involves the intention to offend, or a propositional
content that the speaker intended the hearer to come to believe. The Piss Christ example is much more clearly a case of blasphemy than the
cases I described in the introduction to my chapter because it appears to involve
intention. It is also interesting that apologists for Serrano, and Serrano himself,
appealed to the artist’s intention in order to argue that it was not blasphemous. Definitions of blasphemy Given that we are dealing with a phenomenon
of offence that has broad agreement across different religions, and is of concern
in relation to political issues such as how we should relate to other people’s
religious beliefs in a multicultural society, we require a less culturally specific,
and less religiously specific, definition than the intentional account given by
Perret. But even if intention should not appear as part of the definition of blasphemy,
Aunt Millie’s lack of appreciation of the parrot’s lack of intent is also significant. If intention matters, it may not be the case that it defines whether something was
blasphemous, but the level of culpability. We might appeal to the doctrine of
double effect: it might be thought that the blasphemy might be permissible, or
at least more acceptable, if the blasphemy is the ‘unintended side effect’ of a
good intention, such as the liberation of mixed up human beings from the tyranny 40 Madonna and piano accordion: Disrupting the order of the world of apostles of purity, or the education of the general population of the suffering
of Christ. In suggesting this use of the doctrine of double effect, I suggest that
intention may play a different role from that argued by Perret. Rather than a
definition of the act, intention may play a role in our understanding of the moral
significance of the act. Similarly, while thinking killing someone is wrong prima
facie, we distinguish between different acts on the basis of the killer’s intention. For instance, we distinguish between calculated murder, crimes of passion,
manslaughter, reckless disregard of other people’s safety, killing in war, and
justifiable self defence. These distinctions rely at least in part on the intentions
of the killer. Similarly, in cases of blasphemy, we may refer to cases where we
think it justified (such as the exposure of hypocrisy in the Church, or the
exploration of sincere, but heretical, belief) and ‘gratuitous’ acts of offensive
behaviour.14 Fisher and Ramsay appear to agree with Hoffman that the cultural context
within which the objects are created provides conventional and appropriate
responses for how artists may depict certain images, and for what may count as
blasphemy. Their argument that Serrano’s Piss Christ is unacceptable depends
on a conventional account of what it means to immerse a crucifix in urine. Definitions of blasphemy It
‘can only be a profanation according to the standards of the culture and religion
of which it is an artefact, and photographing it and displaying such a deed can
only be a blasphemy in that culture’.15 This may be made as a stronger, universal claim about defilement. In her
classic text Purity and Danger, Mary Douglas makes some general, and widely
accepted as important, observations about the nature of defilement and its relation
to the manner in which we comprehend the world, and organise our societies.16
However, the attempt to maintain purity and to avoid defilement should not be
seen as a desire that is necessarily totalitarian or fundamentalist. It is something
that is part of everyday consciousness. Dirt, she claims, is matter out of place. The coffee in my cup is clean, but if I spill it on clothes, I am dirty. According
to Douglas, ‘dirt is essentially disorder…Eliminating it is not a negative
movement, but a positive effort to organise the environment’.17 And she claims
that this is not only true of Western secular societies: If this is so with our separating, tidying and purifying, we should
interpret primitive purification and prophylaxis in the same light…rituals
of purity and impurity create unity in experience. So far from being
aberrations from the central project of religion, they are positive
contributions to atonement. By their means, symbolic patterns are worked
out and publicly displayed. Within these patterns disparate elements
are related and disparate experience is given meaning.18 According to Douglas, the categorisation of purity and danger is a major
structuring organisation of our society — representing social hierarchy, 41 Negotiating the Sacred II differentiating between groups within and between societies and cultures, but
also of structuring our relationship with other people in our societies — and
extends beyond human life to the order of the cosmos. We fit into a broader
pattern of relationships. Keeping the purity rules therefore maintains a pattern
or order to the world. If Douglas is correct, the concept of defilement and purity
will be central to all societies, as well as all religions. In summary, it appears that blasphemy is connected to sacrilege, as what we
do with objects, and how we represent them may both involve defilement. The
idea of defilement is cross-cultural, and not merely associated with fundamentalist
or totalitarian religions. Definitions of blasphemy Our intentions do not define whether something is
blasphemous, as what counts as the defilement of an object or symbol is
dependent on socially agreed norms. It is still not clear, however, that the
Madonna and piano accordion is an act of defilement, or that the display of a
Zuni god is an example of such contempt. This is what makes them such
interesting cases. There is nothing clearly offensive about them. In order to
explore what is wrong with such acts, or at least what is perceived to be wrong
about such acts, I explore the cognitive content involved in recognising certain
kinds of symbols. Rules of depiction and the order of the world The connection between how we may represent a symbol, and how we should
respond to one, are linked in Christian tradition. To explain this connection, let
me first address the depiction of Mary, and how she may be depicted in icons. Then let me turn to how she is used within religious worship. Mary holds a special place within Greek Orthodox worship. She was
Theotokos. The title, Theotokos, does not simply mean ‘Mother of God’, but more
precisely ‘the one who gave birth to the One who is God’.19 According to John
of Damascus, ‘The name [Theotokos] in truth signifies the one subsistence and
the two natures and the modes of generation of our Lord Jesus Christ’.20 As
‘the one who gave birth to the One who is God’, she is represented not only as
human, but also as divine.21 Within the Western Church, Mary was not
considered divine, but as being conferred a special grace by God: she was free
from original sin.22 As Theotokos, Mary, as well as her icon, was the legitimate subject of Orthodox
veneration, but under challenge from iconoclasts, the nature of this veneration
needed clarification. Because of her status, Mary has a special place in the
distinction between the adoration, latreia, which was reserved for God, and
reverence, douleia. Thomas Aquinas thought this distinction did not do full
justice to the special position of the Theotokos: she was less than God, but more
than an ordinary human being and more than any saint; therefore, she was not 42 Madonna and piano accordion: Disrupting the order of the world entitled to lateria, but she was entitled to more than dulia. She was entitled to
hyerdulia. 23 While many contemporary artists may consider the signifier as distinct from
the signified, that is, the mental concept may be considered distinct from the
material aspect in which it is represented, no similar distinction is made in
relation to icons in the Orthodox tradition. Within the Orthodox tradition, the
arbitrary nature of the relationship between icon as a symbol and what it
represents is denied. In the icon, the image and the idea are inseparable.24
George Galavaris writes, Based on Neo-Platonic theories, the icon is thought basically a mystery,
a vehicle for divine power and grace, a means of God’s knowledge. It is
not merely a symbol of the archetype, but the represented becomes
present through the icon. Rules of depiction and the order of the world The Council of Nicea of 787 declared that at
the adoration of the icon of Christ the faithful should say, ‘this is Christ
the Son of God’…The icon participates in the holiness of the
represented.25 Accordingly, the honour displayed to the icon is not directed toward the material
object, but to what it represents. It seems that by exploring Platonic ideas we
might understand one way in which an image may be understood as a means of
spiritual knowledge, and how an image may be considered to participate in a
supernatural world. Plato’s understanding of art rests on idealistic assumptions that are first
outlined by the Pythagoreans. According to Liberato Santoro-Brienza, for the
Pythagoreans, art is a ‘therapy of the soul and the body, as a path to attunement
with the secret harmony of the universe, and as a key to decode the order of
reality’.26 The Pythagoreans believed that the entire universe is ruled by
mathematical laws and the connection between the cosmic order, numerical
proportions and musical harmony provided a conception of beauty in terms of
order, measure, proportion and harmony.27 Similarly, for Plato, the ground of
reality is presupposed by any human activity, and guides artistic activity. Empirical reality is a copy of the true reality that is constituted by perfect forms,
and the artist imitates empirical reality.28 It follows from this that the arts are
inevitably deceptive, as they are twice removed from reality, yet the artist may
also be elevated, by inspiration, into the realms of divine harmony.29 Philosophers of the Middle Ages developed and elaborated on Greek
philosophy, but abandoned the Platonic assumption that art is fundamentally
mimetic of empirical reality. Plotinus wrote: ‘The arts do not simply copy visible
objects, but reach out to principles of nature; the arts provide much themselves,
for they can add where there is deficiency; they can do so since in themselves
they possess beauty’.30 Art, on this understanding, is superior to nature. God 43 Negotiating the Sacred II was thought of as the supreme artist: the whole cosmos is a work of art, and the
artist was conceived of as a kind of creator, emulating God’s activity. Rules of depiction and the order of the world The artist’s
task was to transform matter into form, and nature into spirit: it was the
spiritualisation of matter.31 The arts, therefore, have a cognitive, rather than
expressive, character; they involve the recognition and recreation of divine
order, and art works also enable others to understand the world, as artists
produce tokens and symbols of divine perfection.32 From this discussion of Medieval conceptions of beauty, Santoro-Brienza
reconstructs a picture of what is involved in the experience of beautiful things: In the presence of beauty, our senses perceive sensory properties that
stimulate and gratify the sensory functions. Furthermore, in their
immediate apprehension of physical properties, the sensory experiences
lead to an intellectual insight into the structure, order, and form of the
object, so that the intellect is, in turn, stimulated and gratified by an
intuitive grasp of the form… In the harmonious correspondence of the
object to all our faculties and, in turn, all our faculties to the object, our
senses, intellect, our will are satisfied and result in a condition of
delight…The aesthetic experience is, in other words, a harmonious and
unifying experience of immediate and intuitive fusion in which subject
and object cannot be separated and distinguished. That experience is
akin to the experience of love, both physical and spiritual, and of deep
reflection, prayer, and the anticipated beatific vision.33 In the experience of an image, therefore, we may participate in the nature of the
universe and experience unity with it. In Medieval thought, everything participates, in various degrees, with the
perfection of being and, because of this, the different entities share a fundamental
trait or a kind of unity. Santoro-Brienza states: ‘It is in the light of this principle
that we fully understand the Medieval preoccupation with symbolism in general
and the conviction that everything is linked to everything else…being in general
is one, true good.’34 However, the Platonic idea that the arts could be deceptive,
and fail in this task of spiritualising matter, remained important. Augustine
argued that works of art must be partially false insofar as they are the result of
imaginative fiction.35 There is a dual moral aspect to artistic production. Beauty
may align the will and the intellect in desire for and recognition of the form,
but it may also mislead. Symbols, therefore, must be reproduced correctly, as
they have a devotional purpose. Rules of depiction and the order of the world Within the Greek Orthodox tradition, the liturgical arts are one of the ways
in which man becomes divine by grace, and the icon is one of these arts. The
prototype of the image belongs to the corrupt world, but its transcendental 44 Madonna and piano accordion: Disrupting the order of the world quality can be expressed by the fixation of the type from all that is ephemeral. ‘This’, Galavaris states, is why icons cannot be painted according to the imagination of the artists
or a living model. The relationship between the prototype and the image
would have been lost. For this reason the icon-painter uses
manuals…which describe the iconographic scenes and colours to be
used. But the use of manuals alone does not guarantee the painting of
the sacred image. The painter must himself be “illuminated”.36 In the Painter’s Manual, Dionysius of Fourna tells his pupils that they must not
‘carry out this work haphazardly, but with the fear of God and with the
veneration due to a sacred task’.37 The technical skill is only part of this task,
as the icon painter is engaged in a work that externalises spiritual reality. The
true iconographer is engaged in a work of prayer, so that what he creates stems
both from holy tradition and the artist’s own experience of the work of grace.38 The painting of an icon, therefore, may be seen as a form of meditation, and
the manifestation of ‘the other’ that that subject ‘participates in’, or makes
manifest. Its contemplation is a means of revelation, and provides a means for
an audience to engage with this other. The icon used for the poster of the
Adelaide Arts Festival is obviously of Mary. I believe that one may safely assume
that the painter of the Madonna with a piano accordion was not using a manual. Moreover, I assume that the painter of the Madonna with a piano accordion was
not illuminated, let alone offering each stroke of the brush in prayer. The
Madonna is out of context — the syntax of the painting is wrong. She is
generalised, a symbol of a religious art form among other symbols of religious
art. She does not require veneration; she is one among many religious figures. Rules of depiction and the order of the world Given this context, we may understand why this image may be considered to
be connected to blasphemy, as blasphemy is connected to honour due to the
divine or sacred, and the poster subverts or misrepresents what the Greek
Orthodox Church takes to be the nature of the icon, and of Mary. But can we, through this discussion, also understand a religious tradition
that is alien to us, such as the Zuni reaction to the display of their war god? There are obvious, and serious, difficulties. One is that the entire Platonic
tradition of associating beauty and art is dissimilar to many indigenous
conceptions of beauty, and the role of beauty in the production of indigenous
artefacts. The anthropological evidence from other cultures, and common sense,
should stop us from generalising across cultures on the role of aesthetic
experience here. Yet there are some aspects of many indigenous claims and Platonic thought
about the metaphysical nature of representations that appear to be similar. One
is the association of the object with what it represents in such a way that it makes 45 Negotiating the Sacred II little sense to say that something is ‘a representation’. For instance, I have heard
it said that when Maori see images of the ancestors, they do not view or respond
to the image as a representation, but respond to the image by greeting the
ancestors. The idea that the image and what is represented, or the concept and
image, cannot be separated appears to be common to many indigenous concepts
of religious objects and to Orthodox understandings of icons. I do not mean to
imply that Greek Orthodox metaphysics of symbols are the same as indigenous
ones (the Eastern metaphysics of icons is not even the same as the metaphysics
of the Western Christian church); rather, my point is that the Orthodox and
some indigenous metaphysics of symbols may be contrasted to an understanding
of a symbol as a ‘mere’ symbol. Another aspect of this similarity that appears
highly significant in the current discussion is the normative aspects of behaviour
that follow from the metaphysical aspects of the symbol, as religious traditions
involve norms about how an object may be treated, as well as how it may be
produced. There are norms of behaviour for relating to religious objects, and these
norms express the relationship we have with them. Rules of depiction and the order of the world For instance, the icon does
not merely represent Mary; the image and the idea are considered inseparable. Because of this metaphysical relationship, the honour that is due to the icon is
hyerdulia. The acknowledgment within a ritual or religious context is physical. A priest may kiss the icon in recognition. People light candles to the Virgin,
cross themselves and genuflect. Similarly, there are norms of remorse in response
to blasphemy: the tearing and rending of garments.39 Again we find there are
norms of responses to indigenous religious objects. The Maori may greet certain
objects. Other objects, such as the false face masks of the Iroquios, should not
be viewed except within certain contexts. The norms of acknowledgment are not empty gestures, but are elements of
cognition and classification. Nelson Goodman calls these gestures labels. He
writes: ‘Nods of agreement or dissent, salutes, bows, pointings, serve as labels…
The same is true of such activities in response to music as foot- and
finger-tappings, head-boppings, and various other motions. That these are called
forth by the music, while the conductor’s gestures call it forth, does not affect
their status as labels; for labels may be used to record or to prescribe —
“strawberry”, “raspberry”, “lemon”, and “lime” may tell us what is in or what
to put in the several containers.’40 The significance of our responses to music,
Goodman thinks, is their role in analysing, organising and registering what we
hear.41 This relationship between action and concept formation is also held by
philosophers who are opposed to a rationalistic understanding of language. D. Z. Phillips draws this point out in his discussion of Wittgenstein and religious
observance. I will come back to a discussion of religious observance shortly; for 46 Madonna and piano accordion: Disrupting the order of the world the present I wish to focus on the connection between action and categorisation. According to Wittgenstein, religion is a language game, and its basis is not in
conceptual understanding but in our responses to certain symbols and sensory
experiences. For Wittgenstein, ‘primitive reactions’, that is, our reactions to
pain, colours or sounds, are fundamental to language development. The reactions
of jumping with fright, calling colours light or dark or sounds loud or soft,
crying out in pain or expressing shock at the pain of others are fundamental to
the development of concepts. Rules of depiction and the order of the world Wittgenstein wrote: ‘The origin and the primitive
form of the language-game is a reaction; only from this can the more complicated
forms grow. Language…is a refinement; “in the beginning was the deed”.42 Douglas’s account of dirt avoidance similarly links sensory impression and
the recognition of categories that are expressed in labelling gestures, and explains
the process by which those categories are developed and are involved in the
recognition of value. Douglas states: ‘perceiving is not a matter of passively
allowing an organ — say of sight or hearing — to receive a ready-made
impression from without…As perceivers we select from all the stimuli falling
on our senses only those which interest us, and our interests are governed by a
pattern-making tendency, sometimes called schema…’.43 These schemas are
learned, first as response, and then named. These names act as labels. Douglas
writes: ‘Their names then affect the way [sensory impressions] are perceived
next time: once labelled they are more speedily slotted into pigeon-holes in
future.’44 Dirt provides an example of how these categories are ordered into a
normative pattern of schema. Dirt is a residual category, that which is rejected
from our pattern or scheme of classifications — it is out of place. There is a broader generalisation that may be drawn from these observations
that actions are cognitive labels. If actions are cognitive labels, then in making
certain physical responses to religious images we are classifying or categorising
them. But in failing to respond, we may be failing to categorise or to acknowledge
the honour that is due to something as well. Let me give you an example of a
non-religious, conventional response to emphasise this point about the
withholding of a gesture as a display of a lack of respect. After a musical
performance, it is conventionally accepted that clapping hands is an appropriate
form of acknowledgment of the performers’ skill. Withholding this gesture, or
clapping weakly, is a way of expressing dislike. For the performer, a lack of
applause must be devastating. The difference between this kind of labelling of music and the conventional
ways in which we pay respect to something and the form of ritualistic response
that can be seen in acts of devotion, is that in acts of devotion we also recognise
a hierarchy of value and the relationship between the object and observer. Rules of depiction and the order of the world Ritualistic or normative responses are not merely the recognition and classification
of icons and other religious subjects; they acknowledge a relationship between 47 Negotiating the Sacred II ourselves and what we have classified or recognised through the gesture, and
to withhold the gesture may be considered a form of disrespect, or a failure to
apprehend its meaning. The Zuni demand their war gods returned because they
should not be looked at. Clifford tells us that they are considered terrifying
images of great power; as is appropriate to an image of war. Moreover, we can
imagine that the properties used to present these gods exemplify the terror of
war. It is not surprising to find people averting their gaze at certain horrors, or
thinking it would be inappropriate to look upon certain things, such as the
horror of war, with the same disinterested appreciation with which they gaze
at art. Even if gazing upon objects such as sacred masks was ‘an appropriate’
response, indigenous groups might well resent their display. The Grand Council
of Hodenosaunee for instance, has a policy of forbidding the display of medicine
masks that makes a connection between respect for the masks, and the well-being
of the community. The policy states: The exhibition of masks by museums does not serve to enlighten the
public regarding the culture of Hodenesaunee, as such an exhibition
violates the intended purpose of the mask and contributes to the
desecration of the sacred image.45 A claim about the relationship between the ‘order’ of communities and the
treatment of the objects is made explicit. Ruth Phillips reports: ‘The statement
requests the return of masks to the ‘proper caretakers among the Hodenosaunee’
because ‘it is only through these actions that the traditional culture will remain
strong and peace be restored to our communities’.46 This involves a further element of these rules about how we represent, and
acknowledge images or objects that is important in respect to how we
comprehend what might be wrong with the failure to show respect, at least from
the believer’s perspective. This may concern the role of such acts in upholding
or maintaining the order of the universe. Douglas refers to this as a primitive
world view, but it might also be thought of in terms of God’s providence. Rules of depiction and the order of the world This
perspective seems alien to contemporary, secular scholars, and is thought to be
based on mistaken beliefs about the metaphysical relationship between humans
and the world in which they live, and the nature of causation. However, as I
will argue in the next section, we need not accept either belief, and yet still
behave in certain ways that acknowledge respect for certain kinds of
representations. The order of the universe Douglas characterises what she calls ‘the primitive world view’ as a man-centred
universe: ‘A primitive world view looks out on a universe which is personal in
several different senses. Physical forces are thought of as interwoven with the 48 Madonna and piano accordion: Disrupting the order of the world lives of persons. Things are not completely distinguished from persons and
persons are not completely distinguished from their physical environment. The
universe…discerns the social order and intervenes to uphold it’.47 This is
because the transforming energy of the universe is ‘threaded onto the lives of
individuals so that nothing happens in the way of storms, sickness, blights or
droughts except in virtue of these personal links’.48 The universe is capable of
discerning disorder in social relations and social codes, such as whether partners
in sexual relationships are related within prohibited degrees, or whether a person
has murdered a fellow tribesman or a stranger, and an individual’s hidden
emotions.49 There is a connection between the individual and the world in terms
of an order in the world, but this order needs to be maintained. It requires
maintenance through the maintenance of purity rules, but also in terms of the
structure of relationships. Such ideas are also found in the history of Western religious thought. We
find a similar pattern of thought expressed in the speech Shakespeare gives
Ulysses in Troilus and Cressida. The order of the universe includes the orbits of
the planets to the season to the offices and custom of societies. When this becomes
unbalanced, and when things are not in their order, chaos and calamity follow. What maintains the cosmic order for the Elizabethan is the recognition of degree
or value, and the maintenance of social order. Each person and thing has its
place, and its place must be recognised and acknowledged.50 It would be tempting for a non-believer to interpret the need to maintain
order as simply a false understanding of the nature of the universe and of the
causal relations in it. In particular, it might be thought, the causal understanding
of the relationship between failing to show respect to a symbol and social or
natural calamity is false. But, we need not think that a belief in a simple causal
relationship is at work here, and this may be the wrong way to understand the
nature of religious beliefs and gestures of acknowledgment. The order of the universe Phillips thinks that we do not need to believe in the order of reality as a chain
of being, or to hold a particular metaphysical idea about pictures, to think we
would be concerned about doing certain things to images. He agrees with Searle
that one of the most powerful aspect’s of Wittgenstein’s On Certainty is its attack
on the philosophical tradition ‘according to which all our meaningful activities
must be the product of some inner theory…Wittgenstein points out that for a
great deal of our behaviour, we just do it’.51 The explanations should be
considered not to be theories explicitly held, but as means of explicating what
is believed after the act has taken place. He supports this with an empirical experiment, inspired by the idea of ‘acting
out’ a wish suggested by H. O. Mounce.52 Mounce supposed that it would be
hard for almost anyone to comply with a request that they stick pins into the
eyes of a drawing of their mother; but supposing one did do it, and shortly after 49 Negotiating the Sacred II one’s mother developed an affliction in the eye and was in danger of going blind,
then it would be difficult to resist the feeling, if only momentarily, that there
was some connection between the two events. Phillips tested this by asking a
group of 40 students whether they would be prepared to stick pins in the eyes
of a drawing of their mother, and whether, if their mother developed an affliction
in the eye, they would feel guilt. Fifteen of the students said that they would
have no difficulty sticking a pin in the eye of the drawing, and if their mother
subsequently became ill, they would not feel guilt. Their explanation was that
‘it was only a picture’, and that there could be no causal connection between
the two events. The remaining students said that they would not stick pins in
the picture, but that if they did, they would feel guilt if their mother
subsequently developed an affliction of the eye. But, Phillips suggests, the
students who said this were not positing a causal connection when they said
they would feel there was a connection between the two events: ‘[What] they
meant was something like this: they felt that sticking pins in the picture reduces
serious possibilities to a game; it plays around with things. The order of the universe When the affliction
occurs, an internal relation between “playing around” and the event makes the
guilt understandable.’53 Phillips concludes from this that it is not the case that
the students would refuse to stick pins in a drawing of their mother as a
consequence of beliefs about the causal connections between the drawing and
the outcome, but that a primitive moral response occurs and that people, if asked
to reflect on their response, may reply this way.54 There must be some explanation of why most people would not stick pins
into an image of their mother. Douglas’s explanation of ‘the primitive world
view’ in terms of explicit beliefs about an order of the world that needs to be
maintained if chaos and misfortune are to be avoided is one way of understanding
this connection. But the explanation does not address why Phillips’s students
would not stick pins in the eyes of a picture of their mother. Moreover, if
Douglas’s account of this world view could cover all responses to blasphemy,
we would expect the primary response to blasphemy to be fear, rather than
offence or distress. Yet the fact that most people, and it seems even people without religious
beliefs, would not stick pins into a drawing of their mother can be accounted
for in terms of what people hold dear to them, and the respect in which they
hold their mothers. This is consistent with Douglas’s broader claim about the
association between sensory impressions, schema, and values, and Goodman’s
general claims about our responses being a part of the process of cognition. In
recognising something to be an image of ‘our mother’, we classify such images
under a general concept that determines our relationship to it. An icon of the Virgin Mary may similarly be treated with respect, simply
because she is held dear. Non-religious people should not consider such 50 Madonna and piano accordion: Disrupting the order of the world sentiments to be alien to them. When a non-religious person tears up a
photograph of a person they once loved, their act is one of anger and disrespect;
when they refuse to stick pins in the eyes of an image of their mother, or would
feel momentary guilt if something later happened to their mother, they also are
refusing to treat images as ‘mere pictures’. The order of the universe Similarly, we may turn away from
representations of war in horror, even if we know they are fictional. Our
responses to the image may be understood as recognition of the meaning or
content of the image, its value, and an expression of our relationship with it. Blasphemy and homage My interest in this chapter has been to attempt to gain some kind of insight into
what is ‘wrong’ with the seemingly innocent acts of depicting images without
following the rules of their depiction, as in the image of a Madonna with a piano
accordion, and of viewing depictions of gods and indigenous sacred objects
within a museum context. I have attempted to provide as sympathetic account
of these claims and of the religious symbols involved as possible. Because I am
an atheist, I have not been concerned with an assessment of the truth or adequacy
of these ideas, but with their explication, in an attempt to understand a different
perspective. Nor has my objective been to provide a moral argument in support
of or against claims that these acts are actually wrong. My objective has been
to explore whether there is, as my initial intuition suggested, some similarity
between seemingly very different acts, and blasphemy. My conclusion is that the claims that objects should not be displayed in
museums and that certain images are inappropriate are connected through the
idea that neither act displays an appropriate act of homage or respect to what is
represented by the object or symbol. This feature of homage is also central to
the concept of blasphemy, if blasphemy is defined as showing contempt for God
and religious matters through one’s words, thoughts and actions. There is, of
course, a difference between the acts of showing contempt of other people’s
beliefs or of particular beliefs and values, as may occur in intentional blasphemy,
withholding acknowledgment (for example, refusing to applaud a performance),
and failing to acknowledge because you do not ‘understand’ what something
is. In this third case, absolutely ‘nothing goes on’ in our heads when we do not
genuflect or acknowledge a religious meaning. There is no cognitive process. Just as a parrot may blaspheme without intent, a person may blaspheme without
any recognition of value. Just as there are people who would kill a parrot for
blaspheming, there are people who would stick pins into the image of their
mother, and who cannot understand the perspective of people who would refuse
to do this to an image of their mother. Their mistake is to treat images as ‘mere’
symbols. 51 Negotiating the Sacred II Endnotes 1 To Robert and John, with my thanks for their conversation, their warmth, and for sharing their
ceremonies with me. 1 To Robert and John, with my thanks for their conversation, their warmth, and for sharing their
ceremonies with me. 2 James Clifford, 1988, The Predicament of Culture: Twentieth-Century Ethnography, Literature and Art,
Cambridge (MA): Harvard University Press, p. 209. 3 Memo from Michael K. Foster to Dr Barrie Reynolds, Chief Ethnologist, Canadian Ethnology Service,
10 March 1975, File E7-6R, Canadian Ethnology Service, Canadian Museum of Ethnography. Cited in
Ruth B. Phillips, 2004, ‘Disappearing Acts: Exposure, Enclosure, and Iroquois Masks’, in M.S. Phillips
and G. Schochet (eds.) Questions of Tradition, Toronto: University of Toronto Press, p. 74. 4 Ibid., pp. 74-5. 5 Anthony Fisher and Hayden Ramsey, 2000, ‘Of Art and Blasphemy’, Ethical Theory and Moral Practice,
vol. 3, pp. 137-167, p. 138. 6 Ibid., p. 155. 6 Ibid., p. 155. 7 Ibid., p. 139. 8 7 Ibid., p. 139. 8 8 Ibid., p. 139, note 3. 9 Roy W. Perret, 1987, ‘Blasphemy’, Sophia, vol 26, no. 2, pp. 4-14, at p. 5. 9 Roy W. Perret, 1987, ‘Blasphemy’, Sophia, vol 26, no. 2, pp. 4-14, at p. 5. 10 10 Fisher and Ramsay, op. cit., p. 156. One might question the veracity of this account of Serrano’s on
the basis that Fisher and Ramsay are at points quite sardonic about Serrano’s artistic intentions. 10 Fisher and Ramsay, op. cit., p. 156. One might question the veracity of this account of Serrano’s on
the basis that Fisher and Ramsay are at points quite sardonic about Serrano’s artistic intentions. 11 F. Hoffman, 1989, ‘More on Blasphemy’, Sophia, vol. 28, no. 2, pp. 26-35, at p. 30. 12 Fisher and Ramsay, op. cit., p. 157. 11 F. Hoffman, 1989, ‘More on Blasphemy’, Sophia, vol. 28, no. 2, pp. 26-35, at p. 30. 12 Fisher and Ramsay, op. cit., p. 157. F. Hoffman, 1989, More on Blasphemy , Sophia, vol. 28, no. 2, pp. 26 35, at p. 30. 12 Fisher and Ramsay, op. cit., p. 157. 13 Ibid., p. 28. 14 People may accept the doctrine as a factor in determining moral culpability, but, like Fisher and
Ramsay, remain dubious of the artist’s claims about his or her intention or, like Webster, remain dubious
that the artist’s intention is relevant to the interpretation of the work. p
50 E.M.W. Tillyard, 1974 (1943), The Elizabethan World Picture, Harmondsworth: Penguin, pp. 17-8.
51 John Searle, 1987, ‘Wittgenstein: Dialogue with John Searle’, (in The Great Philosophers, Bryan
McGee, BBC Books, p. 346,) cited in D.Z. Phillips, ‘Searle on Language-Games and Religion’, op. cit., p.
28. 52 D. Z. Phillips, ‘Primitive reactions and the reactions of primitives’ op. cit., p. 115-6.
53 Madonna and piano accordion: Disrupting the order of the world 39 Helen Pringle, 2006, ‘Are we capable of offending God? Taking blasphemy seriously’, in Elizabeth
Burns Coleman and Kevin White (eds.), Negotiating the Sacred: Blasphemy and Sacrilege in a Multicultural
Society, Canberra: ANU E Press, pp. 31-43, at p. 32. 39 Helen Pringle, 2006, ‘Are we capable of offending God? Taking blasphemy seriously’, in Elizabeth
Burns Coleman and Kevin White (eds.), Negotiating the Sacred: Blasphemy and Sacrilege in a Multicultural
Society, Canberra: ANU E Press, pp. 31-43, at p. 32.
40 Nelson Goodman, 1968, Languages of Art, Indianapolis: The Bobbs-Merrill Co. Inc., p. 61.
41 Ibid., p. 62.
42 Ludwig Wittgenstein, Culture and Value, (translated by Peter Winch, 1980, Basil Blackwell, 1980,
p. 31) cited in D.Z. Phillips,1993, “Primitive reactions and the reactions of primitives”, in D.Z. Phillips,
Wittgenstein and Religion, New York: St Martin’s Press, p. 114.
43 Douglas, op. cit., p. 36.
44 Ibid. p. 36.
Madonna and piano accordion: Disrupting the order of the world Endnotes Phillips,
Wittgenstein and Religion, New York: St Martin’s Press, p. 114. 43 Douglas, op. cit., p. 36. 44 Ibid. p. 36. 45 Cited in Ruth Phillips, op. cit., p. 75. 46 Ibid.. 47 Douglas, op. cit., p. 88. 48 Ibid., p. 85. 49 Ibid., p. 87. 50 E.M.W. Tillyard, 1974 (1943), The Elizabethan World Picture, Harmondsworth: Penguin, pp. 17-8. 51 John Searle, 1987, ‘Wittgenstein: Dialogue with John Searle’, (in The Great Philosophers, Bryan
McGee, BBC Books, p. 346,) cited in D.Z. Phillips, ‘Searle on Language-Games and Religion’, op. cit., p. 28. 52 D. Z. Phillips, ‘Primitive reactions and the reactions of primitives’ op. cit., p. 115-6. 53 Ibid., p. 116. 54 Ibid. 39 Helen Pringle, 2006, ‘Are we capable of offending God? Taking blasphemy seriously’, in Elizabeth
Burns Coleman and Kevin White (eds.), Negotiating the Sacred: Blasphemy and Sacrilege in a Multicultural
S
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b
ANU E P
31 43
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32 Society, Canberra: ANU E Press, pp. 31-43, at p. 32. 40 Nelson Goodman, 1968, Languages of Art, Indianapolis: The Bobbs-Merrill Co. Inc., p. 61. 41 Ibid., p. 62. p
42 Ludwig Wittgenstein, Culture and Value, (translated by Peter Winch, 1980, Basil Blackwell, 1980,
p. 31) cited in D.Z. Phillips,1993, “Primitive reactions and the reactions of primitives”, in D.Z. Phillips, 42 Ludwig Wittgenstein, Culture and Value, (translated by Peter Winch, 1980, Basil Blackwell, 1980,
p. 31) cited in D.Z. Phillips,1993, “Primitive reactions and the reactions of primitives”, in D.Z. Phillips,
Wittgenstein and Religion, New York: St Martin’s Press, p. 114. g
g
(
y
p. 31) cited in D.Z. Phillips,1993, “Primitive reactions and the reactions of primitives”, in D.Z. Phillips,
Wittgenstein and Religion, New York: St Martin’s Press, p. 114. p. 31) cited in D.Z. Phillips,1993, Primitive reactions and the reactions of primitives , in D.Z. Phillips,
Wittgenstein and Religion, New York: St Martin’s Press, p. 114. 43 Douglas, op. cit., p. 36. 44 Ibid. p. 36. 45 Cited in Ruth Phillips, op. cit., p. 75. 45 Cited in Ruth Phillips, op. cit., p. 75. 46 Ibid.. 47 Douglas, op. cit., p. 88. 47 Douglas, op. cit., p. 88. 8 48 Ibid., p. 85. 48 Ibid., p. 85. 49 Ibid., p. 87. E.M.W. Tillyard, 1974 (1943), The Elizabethan World Picture, Harmondsworth: Penguin, pp. 17 8. 51 John Searle, 1987, ‘Wittgenstein: Dialogue with John Searle’, (in The Great Philosophers, Bryan
McGee, BBC Books, p. 346,) cited in D.Z. 39 Helen Pringle, 2006, ‘Are we capable of offending God? Taking blasphemy seriously’, in Elizabeth
Burns Coleman and Kevin White (eds.), Negotiating the Sacred: Blasphemy and Sacrilege in a Multicultural
Society, Canberra: ANU E Press, pp. 31-43, at p. 32.
40 N l
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1968 L
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61
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g Endnotes 14 People may accept the doctrine as a factor in determining moral culpability, but, like Fisher and
Ramsay, remain dubious of the artist’s claims about his or her intention or, like Webster, remain dubious
that the artist’s intention is relevant to the interpretation of the work. 15 Fisher and Ramsay, ibid., pp. 157-8. 15 Fisher and Ramsay, ibid., pp. 157-8. 16 Mary Douglas, 1966, Purity and Danger, London: Routledge and Kegan Paul. 16 Mary Douglas, 1966, Purity and Danger, London: Routledge and Kegan Paul 18 Ibid., pp. 3-4. 19 Jaroslav Pelikan, 1990, Igmago Dei: The Byzantine Apologia for Icons, The A.W. Mellon Lectures in
the Fine Arts, 1987, The National Gallery of Art, Washington, D.C.: Princeton University Press, p. 134. 19 Jaroslav Pelikan, 1990, Igmago Dei: The Byzantine Apologia for Icons, The A.W. Mellon Lectures in
h
i
h
i
l
ll
f
hi
i
i
i 20 Ibid., p. 135. p
21 Ibid., p. 142. 22 Ibid., p. 140. p
23 Ibid., p. 139. 24 Ibid., p. 3. p
25 George Galavaris, 1981, The Icon in the Life of the Church, Leiden: E. J. Brill, pp. 3-4. 26 26 Liberato Santoro-Brienza, 2003, ‘Art and Beauty in Antiquity and the Middle Ages’, in Stephen
Davies and Ananta Ch. Sukla (eds.), Art and Essence, Westport, Conneticut: Praeger, p. 56. 27 Ibid
57 p
28 Ibid., p. 58. 29 Ibid., pp. 58-9. 30 Plotinus 1966-88, Enneades, v 8, 1, translated by Liberato Santoro-Brienza, ibid., p. 63. 31 Ibid 32 Aquinas, In De Dev. Nom., IV, lectio 5, translated by Liberato Santoro-Brienza, ibid., p. 70. 33 bid 33 Ibid., p. 70. 34 34 Ibid., p. 69. p
35 Ibid., p. 63. p
36 Galavaris, op. cit., p. 4. 37 Cited in John Baggley, 1987, Doors of Perception: icons and their spiritual significance, London:
Mowbray, p. 55. 38 Mowbray, p. 55. 38 Ibid. Ibid. 52 39 Helen Pringle, 2006, ‘Are we capable of offending God? Taking blasphemy seriously’, in Elizabeth
Burns Coleman and Kevin White (eds.), Negotiating the Sacred: Blasphemy and Sacrilege in a Multicultural
Society, Canberra: ANU E Press, pp. 31-43, at p. 32. 40 Nelson Goodman, 1968, Languages of Art, Indianapolis: The Bobbs-Merrill Co. Inc., p. 61. 41 Ibid., p. 62. 42 Ludwig Wittgenstein, Culture and Value, (translated by Peter Winch, 1980, Basil Blackwell, 1980,
p. 31) cited in D.Z. Phillips,1993, “Primitive reactions and the reactions of primitives”, in D.Z. p
50 E.M.W. Tillyard, 1974 (1943), The Elizabethan World Picture, Harmondsworth: Penguin, pp. 17-8. Ibid., p. 87.
50 E.M.W. Tillyard, 1974 (1943), The Elizabethan World Picture, Harmondsworth: Penguin, pp. 17-8.
51 John Searle, 1987, ‘Wittgenstein: Dialogue with John Searle’, (in The Great Philosophers, Bryan
McGee, BBC Books, p. 346,) cited in D.Z. Phillips, ‘Searle on Language-Games and Religion’, op. cit., p.
28.
52 D. Z. Phillips, ‘Primitive reactions and the reactions of primitives’ op. cit., p. 115-6.
53 Ibid., p. 116.
54 39 Helen Pringle, 2006, ‘Are we capable of offending God? Taking blasphemy seriously’, in Elizabet
Burns Coleman and Kevin White (eds.), Negotiating the Sacred: Blasphemy and Sacrilege in a Multicultu Ludwig Wittgenstein, Culture and Value, (translated by Peter Winch, 1980, Basil Blackwell, 1980,
p. 31) cited in D.Z. Phillips,1993, “Primitive reactions and the reactions of primitives”, in D.Z. Phillips,
Wittgenstein and Religion, New York: St Martin’s Press, p. 114. Endnotes Phillips, ‘Searle on Language-Games and Religion’, op. cit., p. 28. 54 Ibid. 53 53 The paintings Leonardo da Vinci began the Last Supper in 1495, on the refectory wall of the
Sta Maria della Grazie, a Dominican convent in Milan. Leonardo’s innovation
was capturing the moment when Jesus announces that someone at the table will
betray him this night.5 The revelatory moment is realistically portrayed by
Leonardo, capturing a wave of emotions — such as surprise, angst, anger, sorrow,
and denial. It took Leonardo twelve years to complete this masterpiece, which
has since been continuously restored and reproduced. The Buddha Shakyamuni
thangka is estimated to have been painted between 1050 and 1100. In this
painting, the Buddha’s right hand is touching the earth, calling the earth to bear
witness to his enlightenment.6 Shakyamuni is attended by two standing
bodhisattvas, the bodhisattva of compassion, Avalokitesvara, and the future
Buddha, Maitreya.7 There are two seated monks above the bodhisattvas. On
the top row stand seven past-Buddhas and another version of Maitreya. The
five dhyana or directional Buddhas sit along the bottom.8 This was a significant
painting when it was executed and today is a highly prized item within a private
Western collection. Both paintings were commissioned. Ludovico il Moro, Duke of Milan,
employed Leonardo to paint, sculpt and design various works, the Last Supper
among them. We know the thangka was commissioned because these details are
written in red Tibetan dbu med script on the back.9 The name of the donor is
not recorded. The Last Supper is large, 460 by 880 cm, and was painted directly
onto a wall. The thangka is 47 by 32 centimetres and was painted on cloth, with
supporting textile mountings and dowel rods at both ends. This construction
permitted it to be rolled up and transported or stored. The themes in the two paintings were not unusual. The Last Supper was
created as a fresco in the dining room of a convent. The painting depicts a supper
in progress — breaking bread, sharing olives and wine — an everyday practice. 4
Materialising the sacred Dianne McGowan This chapter illustrates the shifting meanings of sacred/religious objects, in
particular the recent phenomenon of sacred/religious into fine art commodities. This process, however, may lead to concerns about the new ways in which
religious objects are valued. It is often suggested that this process of secularisation
and commodification is a failure to respect the people who created it, and in
some way presents a harm to the object itself. According to the Oxford definition, the sacred belongs to the consecrated
and the religious; to dedicated objects or purpose; and, objects or persons
affiliated with a deity/god or venerated as holy.1 The aim of this chapter is to
reflect on what has made an object sacred in the past, and perhaps discover a
basis to explain what makes an object sacred in today’s predominantly secular
world. I have selected two religious paintings. Both mark crucial transformative
events. One is drawn from Christianity, the Last Supper by Leonardo da Vinci. The other is a Tibetan Buddhist thangka (painting) of Buddha Shakyamuni. 2 I
have chosen these two images to juxtapose their sacredness and how they have
changed, been re-written or appropriated over time. They are also well-known
representations, requiring no introduction.3 Numerous copies and similar
illustrations are on exhibit in art museums and decorate private homes. I argue that the meaning and value of religious objects is not rigid, but is
fluid and open to modification or re-writing, irrespective of governing norms. Both the Last Supper and Buddha Shakyamuni paintings originated within a
religious context but have since been appropriated into the Western fine art
scene.4 At the same time as the meaning and value of objects is re-written,
however, I argue that the museum context provides a reverential context for
their appreciation. These objects have been formed and shaped from physical
materials, yet appear to be invested with another ‘meta’-materiality. This
something is beyond the tactility of pigment, cloth, wood or bronze. It reaches
across corporeal boundaries, beyond the written word and, for many people,
resonates a sacredness. 55 55 Negotiating the Sacred II Sacred meaning Research on the sacred is hampered by the ‘injudicious’ use of the term ‘sacred’,
which is too often conflated with the term ‘religious’. Peter Fingesten states
categorically that ‘religious’ and ‘sacred’ art are ‘neither synonymous nor
interchangeable…[and] these terms should be applied henceforth with greatest
discrimination in order to avoid confusion’.14 Fingesten defined ‘sacred’ art as
objects and symbols, which conformed to religious law and were consecrated
by prayers, rites or rituals. This would include altars, vessels, vestments,
architecture, sculpture and paintings, whereas, ‘religious’ art is produced outside
of religious restrictions.15 Such a definition would mean that the thangka would
be categorised as sacred art, because it was not only consecrated but executed
according to religious conventions. The Last Supper, however, would be
designated religious art, because it was Leonardo’s expression, he constructed
the scene and the actors’ reactions. It was not a reproduction of an officially
sanctioned painting, rather, Leonardo used everyday practice as his inspiration. The thangka is not simply an idol but enjoys a status equivalent to Greek or
Russian Orthodox icons.16 They are sacred manifestations. According to Jackson
and Jackson, thangkas were a crucial medium by which the ‘ideals of Buddhism
were evoked and brought alive. A sacred painting was for the Tibetan a physical
support — in other words an embodiment — of enlightenment’. They also note
that the simple Buddha Shakyamuni image was commonly commissioned for
accumulating merit towards spiritual advancement.17 Wandering Buddhist teachers made use of the portability of thangkas to
illustrate and inculcate religious messages for the nomadic illiterate population.18
The pictures told familiar stories, in the same manner as the stained glass Wandering Buddhist teachers made use of the portability of thangkas to
illustrate and inculcate religious messages for the nomadic illiterate population.18
The pictures told familiar stories, in the same manner as the stained glass
windows, paintings and sculptures of the Christian church. The wrath of God
unleashed upon the fallen sinner or the innocence of a lamb protected by the
shepherd was silently impressed upon a significantly illiterate population. They
were visual reminders of how to conduct their lives properly, under the
omnipresence of an all-seeing God. The angelic forms of heaven and the horrors
of purgatory were all pictured — graphic reminders that life’s actions were
judged at death. The paintings On a religious level, it is at this supper that Jesus initiates his disciples into the
Christian ritual practice of the Holy Eucharist — the consecration of bread and
wine into body and blood.10 The thangka, on the other hand, was a stupa
furnishing for the eleventh century Tibetan lama and translator Gos Lotsava.11
A stupa is an architectural hollow bell-shaped vessel in which are placed highly
valued items considered worthy to be remembrance offerings.12 These offerings
are not travelling companions on an afterlife journey such as is the Egyptian
practice, but sacred objects offered in appreciation of the person now departed. Even though both these were conceived for religious spaces and depict religious
scenes, only the Tibetan painting was consecrated. A lama ritually opens-the-eyes
of a thangka by painting them in, thereby transforming it from a representation
into a potent living deity.13 I have not found a reference to the Last Supper
being consecrated. 56 Materialising the sacred Materialising the sacred ‘Materialising’ the sacred While the depth of understanding of the spiritual world represented by a thangka
or the church windows may have differed between the illiterate and the
aristocracy, the elite also used their access to resources to highlight their higher
spiritual position and access to the sacred. For example, in Florence during the
1400s it was common practice for churches to sell altar naming rights. Prominent
families would engage prestigious artists to create new altars or altar pieces. Nor
was it necessary for a client to commission an artist. Open market stalls sold
sculptures, paintings and elaborate wooden altars. Contemporary sources of the
day describe these objects as fine art — beautiful but expensive. This ready-made
market also catered for bulk purchases, evidenced by the payment of import
duties for 30 statues of Madonna entering Rome in 1450.19 At the time the Last Supper was being painted there were Christian authorities
who were condemning painterly extravagance because they claimed the
worshipper was being distracted by the skills of the artist.20 In 1494 Girolamo
Savonarola decries the contemporary painterly figures in churches. In his words,
they ‘are with such artifice, adornment and virtuosity that they block the light
of God and true contemplation; people do not consider God, but only the artifice
of the figures’.21 The unpretentious images spelt out their message by repeating
familiar iconography, symbols and colour systems, which the uneducated
understood. In contrast, many popular artists of the Renaissance wanted to
display their skills and their patrons wanted to sponsor art as a display of their
power, wealth and prestige.22 For example, Leonardo painted the il Moro family
crest above the Last Supper, to remind the religious fraternity of the wealth and
beneficence of the il Moro family. Likewise, as already noted, a common purpose
for commissioning a thangka was to gain merit. Quite often the donor was
represented as a small figure sitting respectfully in the lower right hand corner.23
The needs of the wealthy and supplicants created a demand for images of Jesus
and Shakyamuni respectively. Arthur Danto notes that art is constantly validated and revaluated by its
activity in the marketplace, either as an original or as a copy.24 Public
presentation and discourse, whether exhibitions, catalogues, advertisements or
tourist souvenirs, such as posters and postcards, negotiates a consensual view
of an objects’ fitness to be recognised and appreciated as art. Sacred meaning Both paintings, therefore, had a specific religious purpose, and it might be
argued that this purpose ‘fixes’ the meaning or significance of the object, in
contrast to the meaning or significance it has as fine art, in which it is considered
primarily as an object with aesthetic and historical value, but also,
problematically, a commodity to be bought and sold. Yet, we find such objects
have always had financial and aesthetic values that displayed the status of the
possessor. 57 57 Negotiating the Sacred II ‘Materialising’ the sacred This process of
reproduction fuels the desire for art as a collector’s item and commodity. Put
crudely, the greater the recognition/reputation, the greater the desire to own.25
The reproduction of images also contributes to a process in which the object is
considered of value in itself, and the image becomes recognised and adapted in
other images. While access to the Last Supper remains restricted, Andre Malraux notes that
its readily available reproductions has ensured that it is entered into our own 58 Materialising the sacred Materialising the sacred personal lexicon, even though we may never visit Milan to see the original.26
Any member of today’s general public would be able to recall many images that
they have never set eyes on. The profusion of printed material, television,
computers and other technological advances has expanded public access to visual
images. With sufficient fame, the image becomes ‘iconic’. In the same fashion as the image of Sakyamuni has become essentialised as
the historic Buddha, the depiction of the Last Supper by Leonardo is accepted
as the authoritative depiction of that Biblical event. However, the recent clean
and repair by restoration artist Pinin Brambilla Barcilon highlights questions
about the authenticity of the Last Supper as we know it today. Just two
generations after the Last Supper was completed, it was declared that the painting
was already ruined and in need of repainting.27 In 1652 a doorway had been
cut into the centre of the Last Supper wall, destroying part of the lower portion,
which included Jesus’ feet. Thankfully, the early popularity of the painting had
spawned copies and they were used as models for later restorations. There has been a constant reproduction over centuries. Today’s artists have
used the Last Supper to incorporate new socio-political configurations, such as
race and gender. Such uses may be controversial. Recently, the Last Supper was
used as a template for a fashion advertisement. It caused an outcry in France
and Italy, where it was first released and was subsequently banned. A French
judge ruled that the display was a ‘gratuitous and aggressive act of intrusion in
people’s innermost belief’. The posters were ordered down. The prosecuting
lawyer stated ‘when you touch on sacred things you create an unbearable moral
violence which is a danger to our children’. The sacralisation of the aesthetic gaze The availability of reproductions of the Last Supper has made it ‘portable’ and,
as such, it has crossed cultural borders. The portability of thangkas guaranteed
that they could be easily stored rather than destroyed. Further, because thangkas
were consecrated, their disposal could only be by ritualised burning.32 Plainly,
the thangka of Buddha Shakyamuni has suffered sacrilege in the sense that it
must have been looted from the stupa in which it was originally deposited. The
looting may have been by rival monastery centuries ago, or a fortune seeker
yesterday. Since then, the thangka has crossed national boundaries and has
become a possession in a Western art collection. Unlike the familiar sacredness of Christian imagery it has taken the West
some time to become familiar with Tibetan art and to understand its sacred
nature. Tibetan images reflected Catholic missionary thought and were considered
the Devil’s idols. In 1676, the first Western traveller to Lhasa, Jesuit Father
Grueber wrote ‘that our Religion had been here to fore profeffed in this place’. But the Devil ‘hath had the malice to transfer and usurp all the other mysteries
of our faith to his own worship’.33 Missionaries were still promoting this concept
into the early twentieth century. The 1903-04 British expedition into Tibet, led
by Major Younghusband witnessed over 400 official mule loads of objects leaving
Tibet (not including personal souvenirs).34 Many of the Tibetan objects were
settled behind glass vitrines in museums or in curio cabinets in private homes,
occasionally appearing in the market place or auction house. During the early
1900s, few Westerners would have thought of the thangkas as sacred. The contemporary world market is preoccupied with age and worth, whether
it is the church, museum or collector valuing their material assets. Museums and
collectors show little interest in collecting modern thangkas by Tibetan Masters
no matter how traditional they may appear.35 Further, contemporary Tibetan
painters are not encouraged within their own communities to be innovative or
employ contemporary Western styles. ‘Materialising’ the sacred In response, the defence argued that
‘the work is a photograph based on a painting, not the bible…It is a way of
showing the place of women in society today, which is a reflection of our
changing values’.28 Christina Toren, reports on the appropriation of Leonardo’s Last Supper by
the Fijians.29 She notes that Jesus and his twelve apostles, as illustrated in the
Last Supper, represent the same traditional social spacing as the Fijian chiefs. Both parties sit above the general public. Both parties share a ritual repast.30
With the introduction of Christianity, the Last Supper has opened a space by
which the sacredness and authority of the ancestors through the hereditary
chiefs remains potent. Many carpet reproductions of the Last Supper hang in
Fijian churches. They were brought back from Lebanon by young Fijian
servicemen serving under the United Nations Interim Forces in Lebanon. The
fact that the carpet is contemporary and is a product from the Holy Lands
transmutes and while re-writing traditional Fijian sacredness, it also reinforces
these sacred values.31 59 Negotiating the Sacred II The sacralisation of the aesthetic gaze This is not a case of potential sacrilege
but a political attempt by the diasporic Tibetan community to appear as the
authentic agents of the sacred wisdom of Tibet’s lost traditions.36 Jean-Hubert
Martin writes: ‘Religious art is valued when it is ancient, and there is general
recognition that it engendered humanity’s greatest masterpieces.’ He adds that
contemporary works are not thought of as authentic because ‘of the nostalgia
for a time before’.37 There is a sense in the Tibetan aesthetic art world that age
reflects the quality of sacredness. However, the rarefying of Tibetan material culture by Western collecting
and exhibiting negotiates another performance space. The very nature of a
museum implies that objects have been detached from their original contexts.38
The thangka is re-imagined and re-written, not as an object of sacred worship
but as a fine art object precious to Western art tastes.39 According to Stephen 60 Materialising the sacred Greenblatt, museum objects can be thought of not so much as material artifacts
but as visual memory, or memorial.40 The objects are then reduced to tokens
of immortality, the relics of a lost past or monuments to the frailty of cultures.41 In museums, fine art historians, expressing values concerning ‘art-for-art’s
sake’ set about coding the ‘real’ value of the object. In an interview, the veteran
Tibetan art curator, Pratapaditya Pal, speaks of the connection between economic
value and its aesthetic qualities. He states that ‘my primary goal is to establish
a yardstick for beauty in Himalayan art. After all, the price of an object is
generally determined by its aesthetic quality’.42 Ivan Gaskell comments that
the collecting emphasis on aesthetics and art history discourse has
decontextualised the objects, reducing them to collectibles. He states that such
discourse is a ‘very effective means of expunging the sacred qualities of
objects’.43 However, I contend that museums have not necessarily expunged
but add a new exterior gloss by relating to the object with Western values. The
cynic might consider this the addition of a purely economic value. The Tibetan
art on view in museums are curated as aesthetic masterpieces. Many of these
items are borrowed from private lenders and therefore are still in the marketplace. As such, it could be said that Western collecting aesthetics has forged a new
Holy Trinity of ‘authenticated masterpiece’ equals ‘good investment’ equals
‘sacredness’. We need not be this cynical. The sacralisation of the aesthetic gaze Museum ‘aesthetic’ values and the nature of
something as ‘sacred’ may not be incompatible. Ivan Gaskell also notes that
aesthetic qualities are often allied with the sacred. John Huntington is of the
same opinion when he writes: ‘The exceptional artworks gathered here reflect
the religious practices that lead to this compassionate, illuminated state of being,
as well as the myriad aesthetic expressions of its attainment.’44 Present day
museums exhibit to the general public a more acceptable version of the sacred
— non-threatening and non-religious.45 Yet, museums have continued the
church tradition of lighting the pilgrim’s path. Along the way, intensified
pin-spots of light in a subdued space announce the next masterpiece. By
presenting their artworks in an outer silence and under intensified light, the
museum removes sensory distractions, thereby intensifying sensorial experiences,
the precursor of sacred experiences.46 Art philosopher Karsten Harris writes:
‘Stepping into a museum or a concert hall we enter an aesthetic church, a sublime
and rather chilly necropolis, stretching back across time…What needs preserving
does so precisely because it has lost its place in our world and must therefore
be given a special place.’47 Prolific publishing of affordable books has opened the world of art to the
general public. The art historian can describe and interpret a painting for the
uninitiated, however, they cannot experience the painting for the viewer. All
artwork needs spectators to decipher and interpret meaning.48 Images need the 61 Negotiating the Sacred II human gaze to give them voice and pre-eminence over other artworks. The
power of the object is created and deployed by the gaze. As such, the image is
appropriated by the social operation of seeing.49 David Freedberg writes that
the power of images ‘cannot be thought of apart from the desires, needs,
projections, and learned expectations of their viewers’.50 David Morgan calls
this ‘the sacred gaze’. He stresses that the sacred gaze ‘designates the particular
configuration of ideas and attitudes, and customs that informs a religious act of
seeing as it occurs within a given cultural and historical setting’.51 He contends
that the sacred gaze is ruled by protocols that demand a particular performance
and response from the viewer. I would argue that it is only in the last 50 years
that the general public has learned to fix the Western constructed sacred gaze
onto Tibetan thangkas, whereas the Last Supper had long been fixed by the
sacred gaze. The sacralisation of the aesthetic gaze It was an important stop on the ‘Grand Tour’ of Europe.52 16 However, unlike the Greek and Russian understanding that icons are (re)produced by the hand of
the spirit, the Tibetans are ‘ordinary’ artisans, generally born into the profession. There is a repeated
idea that strenuous yogi-like practices precede Tibetan painting. Jackson and Jackson, state that this
is a myth perpetuated by the idealism of textual sources, although, there are historical exceptions and
the ritual practice of the ‘day-thangka’. David Jackson and Janice Jackson, 1988, Tibetan Thangka
Painting: Methods and materials, Ithaca: Snow Lion Publications, p. 12. Oleg Tarasov, 2003, Icon and
Devotion: Sacred spaces in Imperial Russia, London: Reaktion Books.
17 Conclusion It is commonplace within some disciplines to suggest that changes of value and
meaning are ethically problematic. This is considered to be a particular problem
for sacred objects. By juxtaposing the two paintings, I believe that I have been able to isolate
certain aspects of what makes manifest an object’s sacredness to audiences at
particular points in time and in specific contexts. Aspects such as religious
subject, physical setting such as a church or museum, painted by a master,
nostalgia, age, aesthetics and monetary values, all appear to have varying roles
in constructing, maintaining or denying an object’s sacredness. However, I still
believe that the essential ingredient still resides in the paintings themselves. They appear to have the ability to bridge civilisations and mediate between
history and mutable traditions. The paintings condense multiple messages and
convey these to the senses in a way that language cannot.53 They appear to be
vehicles with the capacity to configure spiritual power.54 The Last Supper and
Buddha Shakyamuni reach out and materialise something greater than the physical
materials that went into making them; a resonance that has outlived their makers
and their original audiences; and a resonance that has overcome restorations,
reproduction and relocation. Today’s commodity-orientated world has enhanced
the power of these paintings by exposing them to even greater audiences. Buddha
Shakyamuni and the Last Supper rest in our museum-without-walls lexicon
categorised and stored as both sacred and valuable. 62 Materialising the sacred Materialising the sacred 15 Ibid., p.133.
16 Endnotes 1 The Oxford English Dictionary, Oxford: Oxford University Press, 1970, vol. IX, pp. 16 1 The Oxford English Dictionary, Oxford: Oxford University Press, 1970, vol. IX, pp. 16-7 2 I have maintained the title spelling of the catalogue. Shakyamuni can be equally spelt without an ‘h’. Image and description of cat. No. 114 taken from Pratapaditya Pal, 2003, Himalayas An Aesthetic
Adventure, Art Institute of Chicago, Chicago, pp. 174-5. 2 I have maintained the title spelling of the catalogue. Shakyamuni can be equally spelt without an ‘h’. Image and description of cat. No. 114 taken from Pratapaditya Pal, 2003, Himalayas An Aesthetic
Adventure, Art Institute of Chicago, Chicago, pp. 174-5. 3 It also means that for most readers there is no need for a visual prompt. An illustration of public
familiarity is entering the names into ‘image google’. In just 15 seconds over 2000 hits were recorded
for Last Supper and 2500 hits for Buddha Shakyamuni. 3 It also means that for most readers there is no need for a visual prompt. An illustration of public
familiarity is entering the names into ‘image google’. In just 15 seconds over 2000 hits were recorded
for Last Supper and 2500 hits for Buddha Shakyamuni. y
4 James Clifford, 1988, The Predicament of Culture: Twentieth-Century Ethnography, Literature and Art,
Cambridge, Mass: Harvard University Press. 4 James Clifford, 1988, The Predicament of Culture: Twentieth-Century Ethnography, Literature and Art,
Cambridge, Mass: Harvard University Press. 5 The betrayal passage can be found in New Testament, Mark 14:20-1. 6 6 Earth-witnessing mudra (bhumisparsha) is formed by the extended fingers of the hand touching the
ground. It symbolises Shakyamuni’s enlightenment under the bodhi tree. He summoned the earth
goddess, Sthavara to bear witness to his attainment of enlightenment. Robert Beer, 1999, The Encyclopedia
of Tibetan Symbols and Motifs, London: Serindia Publications, p.154. 7 6 Earth-witnessing mudra (bhumisparsha) is formed by the extended fingers of the hand touching the
ground. It symbolises Shakyamuni’s enlightenment under the bodhi tree. He summoned the earth
goddess, Sthavara to bear witness to his attainment of enlightenment. Robert Beer, 1999, The Encyclopedia
of Tibetan Symbols and Motifs, London: Serindia Publications, p.154. 7 The Dalai Lama is said to be the earthly manifestation of Avalokitesvara. e Dalai Lama is said to be the earthly manifestation o 8 The Five Dhyani Buddhas are the heads of five Buddha families. Endnotes They are Vairochana, Akshobhya,
Ratnasambhava, Amitabha and Amoghasiddhi. Each represents a cardinal direction with one in the
centre. They are not historical figures, like Buddha Shakyamuni, but transcendent beings. Each is
associated with a colour, mudra, an animal that supports his throne, an attribute and bija (seed syllable). A good introduction is the website ‘Tibetan Buddhist Teachings on The Mandala of the Five Dhyani
Buddhas’, http://www.tsl.org/Masters/buddhas/dhyani/frintroduction.html. (Viewed September 2006.) 8 The Five Dhyani Buddhas are the heads of five Buddha families. They are Vairochana, Akshobhya,
Ratnasambhava, Amitabha and Amoghasiddhi. Each represents a cardinal direction with one in the
centre. They are not historical figures, like Buddha Shakyamuni, but transcendent beings. Each is
associated with a colour, mudra, an animal that supports his throne, an attribute and bija (seed syllable). A good introduction is the website ‘Tibetan Buddhist Teachings on The Mandala of the Five Dhyani
Buddhas’, http://www.tsl.org/Masters/buddhas/dhyani/frintroduction.html. (Viewed September 2006.) 9 The script is in the form of dbu med (without head), which means it is less cursive than the commonly
used dbu can (with head) script. The Tibetan language was adapted from Brahmi and written down at
the time Buddhism was introduced into Tibet, seventh century. Dbu med first appears in the twelfth
century. The stupa is bell shaped and represents the enlightened mind. 10 9 The script is in the form of dbu med (without head), which means it is less cursive than the commonly
used dbu can (with head) script. The Tibetan language was adapted from Brahmi and written down at
the time Buddhism was introduced into Tibet, seventh century. Dbu med first appears in the twelfth
century. The stupa is bell shaped and represents the enlightened mind. 10 In reference to taking the bread and the wine, Jesus told his twelve disciples: ‘Do this in remembrance
of me’, 1 Cor 11:23-25. 11 Gos Lotsava (c.992-1074), also known as Drogmi Lotsawa, traveled to India and, after many years
of study, returned to Tibet bringing with him instructions of almost 80 major tantras, including the
important Hevajra tantra. See ‘HH the Sakya Trizin Visits North America’, The Snow Lion Newsletter,
http://www.snowlionpub.com/pages/N50_1.html. (Viewed September 2006,) 11 Gos Lotsava (c.992-1074), also known as Drogmi Lotsawa, traveled to India and, after many years
of study, returned to Tibet bringing with him instructions of almost 80 major tantras, including the
important Hevajra tantra. 14 Peter Fingesten, 1951, ‘Toward a New Definition of Religious Art’, College Art Journal, vol. 10 no.
2, pp. 131-146, at p.144.
15 Endnotes Artist Sandro
Botticelli threw his own art onto these pyres. Savonarola is thought of as a precursor to Martin Luther
(1483-1546). Gilbert, 1998, op. cit., p. 413. 22 Ibid., pp. 439, 444. 23 Western painters have often signed their paintings in this same corner. 24 23 Western painters have often signed their paintings in this same corner. 24 24 Arthur Danto, 1989, ‘Critical Reflections’, Artforum International, vol. 28, (September), pp. 132-133,
at p. 133. 25 Howard Becker, 1982, Art worlds, Berkeley: University of California Press, pp. 306-307. 26 26 Andre Malraux, 1974, The Voices of Silence, St Albans: Paladin, p.16. 27 Giorgio Vasari (1511-1574). 28 27 Giorgio Vasari (1511-1574). 28 28 The defense lawyer also noted that the photograph was inspired by the popular Dan Brown’s Da
Vinci Code. Anon., 2005, ‘French Court Bans Christ Advert’, BBC NEWS (online), 11 March,
http://news.bbc.co.uk/2/hi/europe/4337031.stm. (Viewed September 2006.) See ‘Marithe & Francois
Girbaud: Last Supper’ for photograph. http://www.epica-wards.org/assets/epica/2005/finalists/print/images/27020%20%20%20MFGSPRIN.jpg. (Viewed September 2006.) 28 The defense lawyer also noted that the photograph was inspired by the popular Dan Brown’s Da
Vinci Code. Anon., 2005, ‘French Court Bans Christ Advert’, BBC NEWS (online), 11 March, 28 The defense lawyer also noted that the photograph was inspired by the popular Dan Brown’s Da
Vinci Code. Anon., 2005, ‘French Court Bans Christ Advert’, BBC NEWS (online), 11 March,
http://news.bbc.co.uk/2/hi/europe/4337031.stm. (Viewed September 2006.) See ‘Marithe & Francois
Girbaud: Last Supper’ for photograph. Vinci Code. Anon., 2005, ‘French Court Bans Christ Advert’, BBC NEWS (online), 11 March,
http://news.bbc.co.uk/2/hi/europe/4337031.stm. (Viewed September 2006.) See ‘Marithe & Francois
Girbaud: Last Supper’ for photograph. http://www epica wards org/assets/epica/2005/finalists/print/images/27020%20%20%20MFGSPRIN jpg (
)
http://news.bbc.co.uk/2/hi/europe/4337031.stm. (Viewed September 2006.) See ‘Marithe & Francois
Girbaud: Last Supper’ for photograph. pp
p
g p
http://www.epica-wards.org/assets/epica/2005/finalists/print/images/27020%20%20%20MFGSPRIN.jpg. (Viewed September 2006.) 29 Christina Toren, 1988, ‘Making the present, revealing the past: The mutability and continuity of
tradition as process’, Man (n.s.), vol. 23, no. 4, pp. 696-711. 29 Christina Toren, 1988, ‘Making the present, revealing the past: The mutability and continuity of
tradition as process’, Man (n.s.), vol. 23, no. 4, pp. 696-711. 30 The Christian bread and wine ritual to the traditional Fijian practice of kava drinking. Ibid. 31 31 The fact that the Last Supper is a textile is also relevant to Fijian tradition and the ritual importance
of bark cloths. Ibid., p. 711. Endnotes See ‘HH the Sakya Trizin Visits North America’, The Snow Lion Newsletter,
http://www.snowlionpub.com/pages/N50_1.html. (Viewed September 2006,) 12 His Holiness Dilgo Khyentse Rinpoche (1910-1991), the head of the Tibetan Nyingmapa school, noted:
‘When a great teacher passes away, his body is no more, but to indicate that his mind is dwelling forever
in an unchanging way in the dharmakaya, one will erect a stupa as a symbol of the mind of the buddhas.’
See Stupa Information Page, http://www.stupa.org.nz/. (Viewed September 2006.) 13 ‘Ronald Knox, an Englishman who lived in Sri Lanka in the 1660s and 1670s, observed bronze
foundry practices there. Before the eyes of a Buddha image are made, ‘it is not accounted a God, but a
lump of ordinary metal, and thrown about the shop with no more regard than anything else… The Eyes
being formed, it is thence a God.’ David Freedberg et. al., 1994, ‘The Object of Art History’, The Art
Bulletin, vol. 76, no. 3, pp. 394-396 at p. 85. Furthermore, according to Robert Thurman, the Tibetans
believe that the paintings and sculptures of deities are an extension of the deity. Marilyn Rhie and
Robert Thurman, 1991, Wisdom and Compassion: The Sacred Art of Tibet, London: Thames and Hudson,
pp. 36-37. 14 Peter Fingesten, 1951, ‘Toward a New Definition of Religious Art’, College Art Journal, vol. 10 no. 2, pp. 131-146, at p.144. 14 Peter Fingesten, 1951, ‘Toward a New Definition of Religious Art’, College Art Journal, vol. 10 no. 2, pp. 131-146, at p.144. 17 Ibid., pp. 9, 11. 63 Negotiating the Sacred II 18 Ibid., p. 11. See also figure on p. 9 captioned ‘People of Tarap (Dolpo) viewing thangkas during a
religious gathering’. 19 Creighton Gilbert, 1998, ‘What did the Renaissance patron buy?’, Renaissance Quarterly, vol. 51,
no. 2, pp. 392-450, at p. 407. 20 George Wolfe, 2002, ‘Inner Space as Sacred Space: The temple as a metaphor for the mystical
experience’, Cross Currents, vol. 52, no. 3, pp. 400-411. 21 21 Savonarola (1452-1498) was an Italian Dominican priest and leader of Florence from 1494 until his
execution in 1498. He was known for religious reformation, book burning, and the destruction of art
he thought not suitable. In 1497 he set alight the ‘Bonfire of the Vanities’, here he burnt the excesses
of moral permissiveness, such as mirrors, cosmetics, fine dresses and gaming tables. Endnotes 32 Ann Shaftel notes that according to Mingyur Rinpoche the power/spirit of a Tibetan object is only
destroyed when it is either burned or buried. This means that Tibetan objects held by public and private
collections continue to be ritual objects irrespective of who owns them. Mingyur Rinpoche is a venerated
h
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K
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f Tib
B ddhi
P
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teacher and master of the Karma Kagyu lineage of Tibetan Buddhism. Personal communication. 33 Jesuit Father Grueber travel diaries and letters were documented by Athanasius Kircher (ed.), 1677,
China Illustrata, Amstelodami: apud Jacobum á Meurs, pp. 109, 118. 34 Michael Carrington, 2003, ‘Officers, Gentlemen and Thieves: The looting of monasteries during the
1903/4 Younghusband Mission to Tibet’, Modern Asian Studies, vol. 37, no. 1, pp. 81-109, at p. 104. 35 Contemporary thangkas of traditional style are actively commissioned by Western Tibetan Buddhists. 36 Clare Harris, 1999, In the Image of Tibet: Tibetan painting after 1959, London: Reaktion Books. 37 Jean-Hubert Martin, 2003, ‘The World’s Altars and the Contemporary Art Museum’, Museum
International, vol. 55, no. 2, pp. 38-45, at p. 39. 38 Ibid., p .41. 39 For the sake of this argument I have ignored Western Tibetan Buddhist practitioners. 40 Stephen Greenblatt, 1990, ‘Resonance and Wonder’, in Ivan Karp and Steven Lavine (eds) 1990,
Exhibiting Cultures: The poetics and politics of museum display, Washington D.C.: Smithsonian Institut 40 Stephen Greenblatt, 1990, ‘Resonance and Wonder’, in Ivan Karp and Steven Lavine (eds) 1990,
Exhibiting Cultures: The poetics and politics of museum display, Washington D.C.: Smithsonian Institute
pp. 42-56, at p. 46. 41 Bernard Faure, 1998, ‘The Buddhist Icon and the Modern Gaze’, Critical Inquiry, vol. 24, no. 3, pp. 768-813, at p. 774. Also, Greenblatt, 1990, op. cit., pp. 43-4. 42 42 Jane Casey Singer, 2000, ‘Pratapaditya Pal: Champion of the Himalayas’, Cloudband Magazine, July
21, http//:www.cloudband.com/magazine/. (Viewed December 2003.) 43 Ivan Gaskell, 2003, ‘Being True to Artists’, Journal of Aesthetics and Criticism, vol. 61, no. 1, pp. 53-60, at p.150. 64 Materialising the sacred 44 John C. Huntington and Dina Bangdel, 2003, The Circle of Bliss: Buddhist Meditation Art. Chicago:
Serindia, p.19. 45 45 Gaskell,,op. cit., p. 149. 46 Wolfe, op. cit., pp. 400-11. 47 Karsten Harris quoted in Donald Kuspit, 2004, The End of Art, New York: Cambridge University
Press, p. xiv. 54 Titus Burckhardt, 1967, Sacred Art in East and West: Its principles and methods, London: Perennial
Books, p. 7. Section II The second section explores uses of ‘blasphemy’ against secular and religious
sacred symbols as forms of political protest, or as a force of liberation. ‘The
sacred’ may be understood as a hierarchical structure of symbols that constrain
thought through maintaining stereotypes and social systems of values. In this
sense, what is sacred may be understood in both secular or religious terms, and
its transgression as a means of undermining this hierarchy. This liberating force
is widely accepted by theorists. Herbert Marcuse hails art as a liberating
experience of ‘aesthetic sublimation’ that ‘removes the audience from immediate
engagement with reality’ and ‘subverts the institutionalization of instrumental
reason’.1 In his 1993 book, Blasphemy, David Lawton opines that blasphemy in
the arts is healthy because it ‘often registers the irruption of a new reading
community’2 and marks such a community’s rite of passage. The issue of
blasphemy normally arises in a community that is divided, and it generally arises
because the community is divided. The arts provide a transgressive context in
which artists can challenge established social structures and ideological agendas. Even moderate voices, those less inclined to celebrate or condone all forms of
blasphemy seem prepared to accept this role. Peter Arnds, a literary critic, examines some of the most influential books of
the twentieth century and their subversive use of blasphemy and sacrilege in
order to attack the serious official discourse of totalitarian governments and
preserve the spirit of democracy at times when it is threatened or even disappears. His project hinges on the philosophical dichotomy between rationalism and
irrationalism, that pivotal conflict within modernity and its enfants terribles —
Nazism, Stalinism, and Colonialism. Grass’s The Tin Drum is an allegorical critique
of the Nazi ideology of race and eugenics. Grass’s most blasphemous scenes are
levied against the Church in Germany, specifically its silence during the Nazi
period. Bulgakov’s The Master and Margarita is a grotesque retelling of Christ’s
ascent to Golgotha and his execution, written during the dark era of the Stalinist
purges. To Arnds, it is a ‘novel about the evils of Stalin’s Russia and as such a
monument to the indestructibility of art’ which ‘revives what the Nazis had
tried to eradicate as degenerate’. Both novels parody God and install Satan as
the central figure. Endnotes 48 Karen Stone, 2003, Image and Spirit: Finding meaning in visual art, London: Darton, Longman and
Todd, p12. 49 David Morgan, 2005, The Sacred Gaze: Religious visual culture in theory and practice, Berkeley:
University of California Press, p. 232. 50 David Freedberg, 1994, ‘The Object of Art History’, The Art Bulletin, vol. 76, no. 3, pp. 394-396, at
p. 395. 51 51 Morgan, op. cit., p. 3. 52 52 The Grand Tour was popular from the 1660s to the 1820s and was a tour of European cultural venues. It was an educational rite of passage for wealthy-born bachelors. A grand tour could last from several
months to several years. 53 53 Stone, op. cit., p. 102. 53 Stone, op. cit., p. 102. , p
, p
54 Titus Burckhardt, 1967, Sacred Art in East and West: Its principles and methods, London: Perennial
Books, p. 7. p
p
54 Titus Burckhardt, 1967, Sacred Art in East and West: Its principles and methods, London: Perennial
Books, p. 7. 65 65 1 H. Marcuse, 1978, The Aesthetic Dimension, Boston: Beacon Press, p. 7.
2 David Lawton, 1993, Blasphemy, Philadelphia: University of Pennsylvania Press, p. 139. Section II Rushdie’s Midnight’s Children is secular blasphemy — it
subverts the official view of Indian history as a success story, reinstalls ‘low’
culture over ‘high’ culture, and elevates the marginalised over those at the
centres of power. As in Grass’s novel, the subversion of an official discourse is
achieved specifically through the sacrilegious conflation of important historical
events with the banality of the protagonist’s private life and through a revival 67 Negotiating the Sacred II of myth and irrationalism suppressed by the new state, under Nehru’s politics
of secularisation. In Satanic Verses, Rushdie indulges in what Arnds calls ‘the
blasphemous mixing of religious icons’. The mixing of Islamic elements with
Hinduism earned Rushdie the ire of both communities, including a fatwa from
the former. In the next chapter in this section, Fernandes-Dias explores the controversy
surrounding Les fees ont soif (1978), a play which is considered in the literary
and in the cultural paradigm as a prominent marker of the post Quiet Revolution
assertion of the feminine identity and the social rupture from religious dogmatism
in Québec. Les fees ont soif roused controversy and was banned because of its
polemical attacks on the Québécoise society, the Church, marriage as an
institution, the judiciary and its blasphemous depiction of the Holy Virgin. In
Fernandes-Dias’s essay, the theatrical stage becomes a transgressive space where
the Holy Virgin, the Housewife and the Whore can explore the oppressive
analogies in their exalted or banalised or denigrated roles and voice their profane
polemical attacks on the existing social hierarchy that has subjugated and stifled
them from being more than just their socially accorded functions. The third chapter in this section reminds us that blasphemy need not be
considered an attack on religion, but can occur within a religious tradition with
the emergence of new interpretations. Carolyn D’Cruz and Glen D’Cruz argue
that the film The Temptation of Christ is not so much a critique of religion, as an
exploration of what it meant for God to give the world his son as a human. Instead of emphasising the divine element of Christ, this film emphasises his
humanity. Here, blasphemy is devotional rather than antagonistic. The chapter
shows that a religion cannot be defined by its orthodoxy, and that orthodoxy
must always be partial. Blasphemy is the means by which religions evolve. Endnotes
1 68 5
Blasphemy and sacrilege in the novel
of magic realism: Grass, Bulgakov,
and Rushdie Peter Arnds Fortunately, at times in which the right to freedom of speech is threatened, there
are artists who remind us of that right. In the face of those telling us that we
ought to stand united behind our political leaders and who want to blacklist
unpatriotic academics, in the face of these, we ought to brandish certain books. Books full of blasphemy and sacrilege reminding us that at times of political and
religious monologism, we need to hear conflicting voices in order to preserve
the spirit of liberalism. Mikhail Bakhtin’s theory of heteroglossia still enforces
this message. According to the Russian critic, in the comic modern novel,
heteroglossia is ‘parodic and aimed sharply and polemically at the official
languages of its given time’.1 Bakhtin challenges the tyranny of unitary
languages of regimes founded on religious, national, cultural, racial, or even
linguistic monologues. What are some of these liberal, liberating books that
make the principle of heteroglossia their own in order to subvert political and
religious monologism? To demonstrate how blasphemy and sacrilege are used
to attack secular and religious ideologies, this article will take a closer look at
three world-renowned authors and their texts: Grass’s The Tin Drum, Bulgakov’s
The Master and Margarita, and Salman Rushdie’s Midnight’s Children. Fortunately, at times in which the right to freedom of speech is threatened, there
are artists who remind us of that right. In the face of those telling us that we
ought to stand united behind our political leaders and who want to blacklist
unpatriotic academics, in the face of these, we ought to brandish certain books. Grass John Irving once called Grass’s The Tin Drum the greatest novel by a living
author. To this day, Grass’s book remains one of the most important works of
literature for the construction of postwar German identity. As a writer, Grass
traditionally sides with the oppressed, with society’s marginalised figures. His
novel is, among other things, a literary treatment of the Nazi ideology of race
and eugenics, resulting in the persecution of asocials as ‘life unworthy of life’,
their extermination in psychiatric institutions in the Third Reich, and their
marginalisation in the Adenauer period. Grass’s Tin Drum is the story of Oskar
Matzerath who refuses to grow up during the Nazi regime. Oskar literally stops
growing at the age of three and through his child-like actions of drumming and
screaming glass to pieces, protests against the adult world that surrounds him. 69 Negotiating the Sacred II After the war, he transforms from a child who does not want to grow into a
grotesquely deformed dwarf, whose hump is symbolic of Germany’s ugliness,
of the burden of history that Germany carries upon its shoulders. The Tin Drum
reflects all those paradigms of the carnival with its potential of blasphemy and
sacrilege that Bakhtin outlines in his monumental study Rabelais and his World. All three authors, Grass, Rabelais, and Bakhtin, react through their works to the
oppressive regimes of their times, the rule of Charles V in Rabelais, Stalinist
Russia in Bakhtin, and the Third Reich in Grass. For Rabelais, Bakhtin, and Grass,
the spirit of carnival with its emphasis on the grotesque signifies the symbolic
destruction of authority and official culture and the assertion of popular renewal. A fascinating parallel between Bakhtin and Grass, one that makes it impossible
to read The Tin Drum without thinking of the Russian critic, is that both writers
critique the official discourse of a regime that appropriates folk culture for its
oppressive politics and for rejecting and killing undesirable individuals. While
Bakhtin uses theory, the theory of blasphemy to attack Stalinism, Grass attacks
German conservatism more directly through fiction. While for obvious reasons,
the more muted criticism of Stalinism by people like Bakhtin and the Russian
novelist Mikhail Bulgakov was levelled entirely at the State, Grass’s most
blasphemous scenes are levelled against the Church in Germany, specifically
against its silence during the Nazi period. Grass The difference between aboriginal peoples and pagan societies, which manage
to wed the forces of chaos with the forces of order, and Christianity, which
started separating these two principles by distinguishing between Jesus and
Satan, becomes interesting within the context of Grass’s novel, which reflects
this pagan union of the Apollonian and Dionysian sphere. It does this by merging
in Oskar Matzerath the figures of Jesus and Satan, of victim and fascist. Oskar
can never be just one: the dividing line of any dialectic is blurred in this book
and, like all archetypal tricksters, Oskar finds himself on the threshold between
two domains. The trickster's typical location in European culture is the
marketplace, and as Bakhtin tells us in the Middle Ages and the Renaissance,
this was the place in which curses, profanities, and oaths reigned.2 While the
sacred was reserved for the church, the profane reigned outside of church,
primarily in the marketplace. Pro-fanum translates into ‘before the temple’. In
early modern European culture, the grotesque became most visible in the
marketplace, a place not only of multicultural interaction but also a venue for
all sorts of groups that later became increasingly segregated from society: gypsies,
transient musicians, exotics of doubtful origin, freed slaves, midgets and giants.3
Yet with the formation of the bourgeois class, socially inferior classes in
marketplaces and fairs increasingly became the ‘object of the respectable gaze’4
by which the bourgeois class was able to confirm its own superiority. Particularly, the slave from the colonies, the dwarf, and the pig were displayed
and celebrated at the fair because of their low status. Alongside with their 70 Blasphemy and sacrilege in the novel of magic realism: Grass, Bulgakov, and Rushdie segregation within the forming nation-state, slaves and dwarves then became
increasingly banished from church.5 Oskar’s presence in church is in itself a
violation of the sacred realm through the profanity of his grotesque body and
all his body stands for, as opposed to one of his counter images, the classical
body of the athlete on the cross, who is flexing his muscles and expanding his
chest over the main altar of the Danzig Sacred Heart Church. In addition, Oskar
repeatedly violates this physical division between the sacred and the profane
by taking profane language and actions into church. Grass As a culture of shame and
guilt, Germany in the 1950s had its areas of silence — the Nazi crimes, the
Holocaust and euthanasia. A central moment of breaking this silence about the
Holocaust, in which Oskar accuses the church, that sacred domain opposed to
the grotesque, of its passivity in the face of Nazi atrocities, occurs when he gives
the Jesus figure his drum and tells him to use it, as a way of protesting against
what is happening on the political stage at the same time. This is a double
disruption of the sacred, both in the sense that Germans in the 1950s did not
want to hear about the Holocaust and the war, and in locality, the desecration
of the sacred ground. The Catholic Church in particular is the target of Grass’s satire. As Günter
Lewy has shown, the Catholic Church silently supported the Final Solution,
while it followed the general public in its outcry against the practice of
euthanasia.6 The main reason for the Catholic Church's protest against euthanasia
was that here Germans were killed while the Final Solution targeted the Jews,
who had killed Jesus Christ.7 In 1939, Archbishop Gröber argued that because
the Jews had killed God, Christianity was not to be regarded as a product of the
Jews but was ‘in the most intimate union with the Germanic spirit’.8 This
appropriation by the German church of Jesus into its own ranks is reflected in
Grass’s Aryanisation of the Jesus figure à la Leni Riefenstahl. Grass’s church
scenes exhibit some of the most offensive passages in the book by conflating
sacred images with what Bakhtin calls the material bodily lower stratum, as
Catholicism never ceases to inspire Oskar with blasphemy. In picaresque fashion,
Oskar mutters commentaries on the Mass while moving his bowels, he equates
Jesus with his philandering father Jan Bronski, he touches the little Jesus figure’s
penis, his watering can, as he calls it, thus giving himself a massive erection,
and he sits on the Virgin Mary’s thigh. In the marketplace, Bakhtin argues, ‘the
most improper and sinful oaths were those invoking the body of the Lord and
its various parts, and these were precisely the oaths most frequently used’.9
Oskar’s drumming, and especially the drumsticks, belong to a series of phallic
symbols. Grass Being dactyls like the Tom Thumb figure himself that Oskar is partly
modelled on, these are grotesque images of potency that contrast starkly with
Jesus’s own flaccid penis as a symbol of the church’s political impotence. These
carnivalesque images subvert the authority of the church, conflating the theme
of the Holocaust with folk humour. 71 Negotiating the Sacred II Negotiating the Sacred II Oskar’s blasphemies turn into crime when he steals nativity figures from
numerous churches. In these later church scenes, Oskar uncrowns the church
Jesus by adopting his name as the leader of a street gang and by sawing him off
along with the other two figures, John the Baptist and the Virgin Mary. He has
his ‘disciples’, the gangsters named the Dusters, perform Catholic rituals such
as genuflections by the holy-water font or enact an impromptu Mass and invoke
the ite missa sunt, a line that was also the object of derision in the medieval Feast
of Fools, where it was converted into the threefold braying of an ass performed
by the priest.10 We can see to what extent Grass’s book from 1959 is steeped
in these early cultural traditions of Europe. In synchrony with other tricksters
who muddy high gods and ‘are made in and for a world of imperfections’, in
which they ‘do not wish away or deny what seems low, dirty, and imperfect’,11
Oskar’s blasphemies in church challenge religious idealism and indict the church’s
silence towards the Nazis’ practice of euthanasia, of the killing of the physically
and mentally degenerate, of doing away with what seemed to them low, dirty,
and imperfect. This union between the church and the totalitarian state is
explicitly addressed in Grass’s equation of the classical body of Jesus and the
perfect Aryan body, the athletes of the 1936 Olympics and Jesus’s blue eyes,
and the equation of the holy cross and the swastika. Oskar questions Jesus as a
culture-hero and concludes that he, Oskar, is a more genuine Jesus than the
other, for at least Oskar drums resistance to the Nazis with his tin drum. His
actions seem to imply the question: Where was God during the Holocaust? Where
were Jesus’s miracles then? Moreover, Oskar’s second form of protest, his desire
to scream glass to pieces in church, could be read as a form of protest against
the broken glass during Reich Crystal Night and the church’s silence. Bulgakov Grass’s book is part of a tightly woven fabric of texts that are steeped in the
tradition of the Menippean satire. The revival of this genre in the twentieth
century, whether in Russia or Germany, has the function of social criticism. It
is subversive, blasphemous and sacrilegious literature that forms a carnivalesque
counter-culture to the seriousness of any officialdom. Bakhtin wrote his book
on Rabelais with this intention and he did for literary theory what Mikhail
Bulgakov’s The Master and Margarita did in fiction. The publication of both
books was delayed for many years due to the censorship under Stalinism. Bakhtin’s dissertation on which his book is based was not completed until 1940,
he did not receive his degree until 1951, and the book was not published until
1965. Bulgakov’s novel experienced a similar publication history: it was started
in 1928 but was not published until 1966, 26 years after his death, as a censored
version in a Moscow journal. Mikhail Bulgakov’s The Master and Margarita is
a novel of grotesque, fantastic realism written during the dark era of the Stalinist
purges, a novel about the evils of Stalin’s Russia and, as such, a monument to 72 Blasphemy and sacrilege in the novel of magic realism: Grass, Bulgakov, and Rushdie the indestructibility of art much like Grass’s novel, which also revives what the
Nazis had tried to eradicate as degenerate. Bulgakov’s satire, too, lends itself to
a Bakhtinian interpretation with its concept of the carnival as a temporary
liberation from the prevailing truth and from the established order. Like Grass’s
novel, The Master and Margarita was itself influenced by Rabelais. Bulgakov’s
novel is about the fate of a novel about Pontius Pilate and Jesus Christ, written
by the protagonist, the Master. It blends a retelling of Christ’s ascent to Golgotha
and his execution with the incredible events surrounding a visit by Woland,
Satan, the central trickster figure in this text, and his minions, to the Moscow
of Stalinism. The presence of Jesus, Ieshua, as a character of a novel was
unthinkable for the atheist Stalinist era. This is very different from Grass’s kind
of blasphemy, a blasphemy within and directed against the Church. Bulgakov’s
novel abounds if not in blasphemy then in a sort of secular sacrilege targeted at
the state and the absence of any church. Bulgakov Evidently, at times of extreme censorship only
metaphorical language can save the artist from persecution; hence the magic
realist novel can be a tool to express and attack the politics of totalitarian regimes. What this novel describes as witchcraft and fantasy, was, however, a grim reality
that had nothing to do with magic. The novel displays a rediscovery of the
iconography of Hell, which had gone missing in the bourgeois age, that joyful
hell, of which Bakhtin speaks, to be found in medieval carnival and in Rabelais. Griboedov House, for example, the primary hangout for the Muscovite literary
world and for Woland and his devils is such a hell. Particularly its famous
restaurant, in which Woland gives his satanic ball, with its incredibly loud jazz
band and the laughter that roars through its vaults is the main carnival locus
which parades beauty and ugliness, life and death, and is reminiscent of Grass’s
restaurant, the Onion Cellar and its own jazz band. In Grass’s Onion Cellar,
Germans are taught to mourn by crying over onions. In both novels jazz has a
liberating function from oppression. In The Tin Drum, Oskar disperses a Nazi
party rally from inside a rostrum by drumming to the rhythm of a Charleston,
‘Jimmy the Tiger’, which has the power to dissolve the uniformity of the saluting
Volkskörper, the folk body. In accordance with Cornel West’s thoughts in his
recent book Democracy Matters on the potential of jazz and blues to resist the
stifling of democracy and to give tragicomic hope in the face of racism, Oskar’s
music temporarily dissolves the rigidity of the Sieg-Heil salute, a central symbol
of the Nazi race ideology. Similarly, the jazz in Bulgakov’s Griboedov House
can be read as a carnivalesque form of resistance to the Puritanism of Stalinism. Like The Tin Drum, The Master and Margarita parodies God, installs Satan
as a central figure, is intertextual with Goethe (Faust in Bulgakov, the Goethean
Bildungsroman in Grass), conflates the rational with the irrational, pits irrational
elements against the rationalist party monologue, cosmopolitanism with
nationalism. Bulgakov’s devil Woland, for example, is described as a polyglot
foreigner. In both novels, the idea of cosmopolitanism clashes with the politics
of rootedness. Primarily, however, Bulgakov’s novel fulfils what Bakhtin saw
in Dostoevsky’s novels, their polyphony, heteroglossia, a never-ending dialogue
in opposition to the monologism of Stalinism, a novel that is open-ended,
unfinalised. Bulgakov In the two central figures, the Master
and Pontius Pilate, we obtain the two positions of the persecuted artist and the
tyrant. The Master is to an extent a self-portrait of Bulgakov and his difficulties
as a writer under Stalin. Stalin can be discerned both in the figure of Pontius
Pilate, who condemns innocent people to death, and Woland, the Devil. The
novel shares with Grass’s The Tin Drum the installation of a vagrant Jesus figure,
who in both cases has to carry the burden imposed upon him by the crimes of
a totalitarian regime, its cowardice, betrayals, and murders. That Oskar Matzerath
increasingly identifies with Jesus at the end of Grass’s novel indicates that he
is the victim of the crimes of humanity par excellence and, like Bulgakov, Grass
intends to remind us that the story of Jesus continues to have metaphorical
relevance for our own day, that there will always be a Pontius Pilate who ends
up crucifying a Jesus. Bulgakov’s novel can quite literally be characterised as a work of magic
realism. It engages in mock uncrownings of officially respected figures from the
world of literature and theatre and makes fun of the mysterious disappearance
of Moscow inhabitants, a common phenomenon under Stalinism. The unreality
of the totalitarian regime, which Hannah Arendt has described as a ‘superior
realism’ surrounding these regimes, may be the reason why some artists respond
to these times with works of magic realism.12 Mikhail Bulgakov, for fear of
retribution, never talks openly about Stalinism but refers to it as witchcraft:
‘And it was two years ago that inexplicable things began happening…people
started disappearing without a trace. Once, on a day off, a policeman appeared,
summoned the second lodger (whose name has been lost) into the front hall, and
said that he had been asked to come down to the police station for a minute in
order to sign something. The lodger told Anfisa…he would be back in ten
minutes…Not only did he not return in ten minutes, he never returned at all…it
was witchcraft pure and simple, and …as everyone knows, once witchcraft gets
started, there is no stopping it’.13 One of the most famous lines from this novel 73 Negotiating the Sacred II — ‘once witchcraft is started, there is no stopping it’ — expresses
totalitarianism’s self-consuming racism, the never-ending process of having to
find enemies. Bulgakov It is passages like this one that mark moments in the text where,
due to the conflation of reality with surrealism, the term ‘magic realism’ becomes
justified in the highest degree. Evidently, at times of extreme censorship only
metaphorical language can save the artist from persecution; hence the magic
realist novel can be a tool to express and attack the politics of totalitarian regimes. What this novel describes as witchcraft and fantasy, was, however, a grim reality
that had nothing to do with magic. The novel displays a rediscovery of the
iconography of Hell, which had gone missing in the bourgeois age, that joyful
hell, of which Bakhtin speaks, to be found in medieval carnival and in Rabelais. Griboedov House, for example, the primary hangout for the Muscovite literary
world and for Woland and his devils is such a hell. Particularly its famous
restaurant, in which Woland gives his satanic ball, with its incredibly loud jazz
band and the laughter that roars through its vaults is the main carnival locus
which parades beauty and ugliness, life and death, and is reminiscent of Grass’s
restaurant, the Onion Cellar and its own jazz band. In Grass’s Onion Cellar,
Germans are taught to mourn by crying over onions. In both novels jazz has a
liberating function from oppression. In The Tin Drum, Oskar disperses a Nazi
party rally from inside a rostrum by drumming to the rhythm of a Charleston,
‘Jimmy the Tiger’, which has the power to dissolve the uniformity of the saluting
Volkskörper, the folk body. In accordance with Cornel West’s thoughts in his
recent book Democracy Matters on the potential of jazz and blues to resist the
stifling of democracy and to give tragicomic hope in the face of racism, Oskar’s
music temporarily dissolves the rigidity of the Sieg-Heil salute, a central symbol
of the Nazi race ideology. Similarly, the jazz in Bulgakov’s Griboedov House
can be read as a carnivalesque form of resistance to the Puritanism of Stalinism. — ‘once witchcraft is started, there is no stopping it’ — expresses
totalitarianism’s self-consuming racism, the never-ending process of having to
find enemies. It is passages like this one that mark moments in the text where,
due to the conflation of reality with surrealism, the term ‘magic realism’ becomes
justified in the highest degree. Rushdie While Grass writes against the political abuse of folk culture in Nazi Germany,
Bulgakov writes against the suppression of the Christian myth following the
Communist revolution. In both books, myth is disentangled from the grasp of
ideological abuse. This also happens in Rushdie’s books, both in Midnight’s
Children and Satanic Verses. As Grass’s The Tin Drum draws on previous texts,
it becomes the palimpsest for later texts. Salman Rushdie openly admitted to
having been influenced by Grass.14 The parallels between these two authors
are legion. While Grass juxtaposes Germans and Poles, one of Rushdie’s central
themes is the relationship between Muslims and Hindus. The works of both
writers display a high degree of intertextuality and heterogeneity and both have
provoked strong reactions from the people who do not agree with their use of
blasphemy and sacrilege, although arguably conservative reactions to Grass’s
The Tin Drum in places like Bavaria in the early 1960s and Oklahoma in the
mid-1990s fade in comparison with the fatwa which Khomeini placed on Rushdie
after the publication of The Satanic Verses in 1989. One of the most interesting
connections between The Tin Drum and Midnight’s Children is their fantastic
realism, their revival of the picaresque tradition. Most of Rushdie’s characters
display the kind of homelessness of which Bakhtin speaks, the dubious origin
of the picaro, that great blasphemer in world literature. We realise to what extent
the picaro himself stems from an intercultural archetype that transcends Europe,
the mythological trickster, whose central function is to criticise society from its
margins. As tricksters, Oskar Matzerath and Saleem Sinai, the hero of Midnight’s
Children, have in common that they are equipped with magic weapons allowing
them to commit deeds of blasphemy and sacrilege. What is Oskar’s scream, by
which he destroys the glass in churches, is Saleem’s extremely sensitive nose
that allows him to smell the thoughts of people. As tricksters, they are
marginalised, figures on the threshold, and find themselves in what the
anthropologist Victor Turner has called a zone of liminality.15 It is precisely
from their liminal position that they are able to levy acts of blasphemy criticising
official ideologies, the times in which they live. Both stand on the threshold
between two historical ages, Nazism and post-war Germany on the one hand,
India as a British colony and postcolonial India on the other. Bulgakov In a key moment, in the Master’s novel within the novel, he has
Yeshua save Pilate from eternal damnation, thus guaranteeing the persistence
of a dialogue between irrationalism in the form of faith (Jesus) and reason (Pilate,
Stalin). This union between rationalism and irrationalism also comes about in
The Tin Drum, where Oskar advocates a union between the Apollonian and the
Dionysian principles borrowed from Nietzsche’s Birth of Tragedy. It seems that 74 Blasphemy and sacrilege in the novel of magic realism: Grass, Bulgakov, and Rushdie these two European novels finally attempt to achieve to mend the central split
that goes through the cultural history of modernity, that split between reason
and unreason that emerges from the Enlightenment. Rushdie Both texts revolve
around the Stunde Null, the zero hour of the nation state, 1945 in Germany and
1947, the day of Indian Independence. 1945 marks the date when Oskar is
transformed into a grotesquely misshapen dwarf, while precisely at the stroke
of midnight preceding Indian independence, Rushdie’s protagonist Saleem Sinai
is born together with the other thousand midnight children. What moves these 75 Negotiating the Sacred II two novels into close proximity is the grotesque body of the protagonist and
their grotesque, sacrilegious representation of history. While Oskar’s hump is
a symbol of German guilt that he carries on his shoulders and while his deformity
also reflects the ugliness of Germany after 1945, Saleem Sinai, whose
physiognomy resembles that of the Hindu Elephant God Ganesha, carries the
very shape of the Indian subcontinent in his face, as his teacher Zagallo points
out with his Peruvian accent: ‘Sons of baboons! Thees object here’ — a tug on my nose — ‘thees is
human geography!’ ‘How sir where sir what sir?’ Zagallo is laughing
now. ‘You don’t see?’he guffaws. ‘In the face of thees ugly ape you don’t
see the whole map of India?’ ‘Yes sir no sir you show us sir!’ ‘See here
— the Decan peninsula hanging down!’ Again ouchmynose. ‘Sir sir if
that’s the map of India what are the stains sir?’ It is Glandy Keith Colaco
feeling bold. Sniggers, titters from my fellows. And Zagallo, taking the
question in his stride: ‘These stains,’ he cries, ‘are Pakistan! Thees
birthmark on the right ear is the East Wing; and thees horrible stained
left cheek, the West! Remember, stupid boys: Pakistan ees a stain on the
face of India!’ ‘Ho ho,’ the class laughs, ‘Absolute master joke, sir!’ But
now my nose has had enough; staging its own, unprompted revolt against
the grasping thumb-and-forefinger, it unleashes a weapon of its own…a
large blob of shining goo emerges from the left nostril, to plop into Mr
Zagallo’s palm. Fat Perce Fishwala yells, ‘Lookit that, sir! The drip from
his nose, sir! Is that supposed to be Ceylon?’16 ‘Sons of baboons! Thees object here’ — a tug on my nose — ‘thees is
human geography!’ ‘How sir where sir what sir?’ Zagallo is laughing
now. ‘You don’t see?’he guffaws. Rushdie ‘In the face of thees ugly ape you don’t
see the whole map of India?’ ‘Yes sir no sir you show us sir!’ ‘See here — the Decan peninsula hanging down!’ Again ouchmynose. ‘Sir sir if
that’s the map of India what are the stains sir?’ It is Glandy Keith Colaco
feeling bold. Sniggers, titters from my fellows. And Zagallo, taking the
question in his stride: ‘These stains,’ he cries, ‘are Pakistan! Thees
birthmark on the right ear is the East Wing; and thees horrible stained
left cheek, the West! Remember, stupid boys: Pakistan ees a stain on the
face of India!’ ‘Ho ho,’ the class laughs, ‘Absolute master joke, sir!’ But
now my nose has had enough; staging its own, unprompted revolt against
the grasping thumb-and-forefinger, it unleashes a weapon of its own…a
large blob of shining goo emerges from the left nostril, to plop into Mr
Zagallo’s palm. Fat Perce Fishwala yells, ‘Lookit that, sir! The drip from
his nose, sir! Is that supposed to be Ceylon?’16 The text abounds in ‘images of the grotesque body draining into the world’
which serve ‘a Menippean vision of renewal and progress’ that is ‘referentially
directed towards the qualities of Indian society that Rushdie valorises: pluralism,
democracy, hybridity, and change’ and they satirically attack the ‘forces in
modern India and Pakistan that deny those principles: fundamentalism,
despotism, purity, and stasis’.17 The representation of history in Midnight’s Children is steeped as much in
folk culture as that of The Tin Drum. In a conversation that Grass and Rushdie
had in the mid-1980s, they both admitted to each other that their use of the
fantastic and of the fairy-tale world stems from their cultures’ individual literary
heritage. While Rushdie emphasises the importance of the Arabian Nights for
his novel, of the stories of A Thousand and One Nights, Grass’s literature is deeply
rooted in the German Baroque and Romantic tradition, the picaresque novel and
the fairy tales. Like Grass, who destabilises the mendacity of post-fascist
rationalism, Rushdie attempts to subvert the official view of Indian history as a
success story, as what Nietzsche called ‘monumental history’, the history of
India’s great leaders.18 Against this monumental vision of India’s history,
Rushdie offers his critical view of history. The story of Saleem Sinai is the story 76 Blasphemy and sacrilege in the novel of magic realism: Grass, Bulgakov, and Rushdie of India’s common people. Rushdie Like Grass, Rushdie thus reinstalls ‘low’ culture over
‘high’ culture, elevates the marginalised over those at the centres of power. The
subversion of an official discourse is, in both cases, achieved specifically through
the sacrilegious conflation of important historical events with the banality of
the protagonist’s private life and through a revival of myth and irrationalism
suppressed by both new states, both under Adenauer’s politics of rationalism
and Nehru’s secularisation. In order to show how ludicrous the concept of the
Stunde Null, the zero hour, this sort of tabula rasa made of Nazism or colonialism,
really is, Rushdie recycles a central scene from Grass, the Onion Cellar episode,
which resurfaces as Mumbai’s ‘Midnite Confidential Club’. Both authors want
to bring back the past and understand that nothing will disappear, everything
will come back. What is catharsis through onions in Grass corresponds to
Rushdie’s chutney, the sweetsour ‘chutney of memory’. Preserving the past is a concern in all three books. Rushdie’s image of the
pickling factory that Saleem works in is a fitting motif for this chutneyfication
of history and the narrator compares his thirty chapters with pickle jars in which
the past is preserved. Grass, Bulgakov, and Rushdie engage in myth in order to
preserve the past and a cultural heritage that the great rulers of their countries
threaten either to manipulate or to suppress. Grass offers his vision of myth and
folk culture in opposition to the Nazis’ ideological manipulation of myth and
folk culture and the spirit of rationalism under Adenauer. By way of parody,
Bulgakov reinstalls central icons of the Christian myth in opposition to Stalin’s
imposed atheism. Rushdie writes in opposition to Jawaharlal Nehru and his
daughter Indira Gandhi’s suppression of Hindu and Muslim traditionalism for
the sake of their monumental vision of history that focuses on the great leaders
but not on the people. In Midnight’s Children, he revives Hindu myth, for
example, in the character of the Widow who drains all hope from the 1001
children born at the stroke of midnight of 15 August 1947, the day of Indian
Independence from colonial rule. Rushdie This figure can serve as a pars pro toto to
demonstrate that this novel is a work of magic realism, for, at a realistic level,
the Widow symbolises Indira Gandhi and her rule of terror during the Emergency
period (1975–77), while at the level of myth, she is made to resemble the evil
goddess Kali, who is often represented ‘with protruding tongue, garland of
skulls, and hands holding weapons and severed heads, stark naked upon the
prostrate body of her beloved consort Shiva’.19 This revival of Hinduism becomes even more problematic in The Satanic
Verses where, in truly satirical manner, elements are shamelessly mixed, in this
case the blasphemous mixing of religious icons. When the Muslim Gibreel, one
of the two central characters, enters the film world, one of his first roles is to
play the Hindu god Ganesha, with elephant trunk and large ears. Later, he
metamorphises into Hanuman, the monkey king from the epic Ramayana. Rushdie’s deconstruction of the dictatorial politics of the Indian film industry 77 Negotiating the Sacred II reminds us of Grass’s and Bulgakov’s parody of monological prose like the
Bildungsroman of German Classicism and Socialist Realism, but among the greatest
offences is Rushdie’s mixing Islam with Hinduism. In conclusion, one could argue that the literature of magic realism attacks
primarily the official discourses of the church and the state. In this satirical
literature, which is steeped in earlier European traditions such as the menippean
satire or the picaresque novel, blasphemy and sacrilege, the rupturing of the
sacred realm (church and state) through the profane (pro-fanum), are levelled
against the Church’s and the State’s mechanisms of oppression. If we believe
with Horkheimer and Adorno in the dialectic of Enlightenment, this implies that
reason can become oppressive, that rationalism can reach a point at which it
perverts into irrationalism, which no doubt it did under totalitarian rule. In the
fiction of magic realism, Western rationalism becomes the target of satirical
representations of irrationalism. What Deleuze and Guattari have, in their book
Mille Plateaux, called the arborescence of Western societies, the deep roots of
their teleologies and their territorialism, is being subverted through what they
call the rhizome, the shallow roots associated with deterritorialisation, nomadism,
and homelessness, for which tricksters, picaros, and other literary nomads are
literary representations. In this body of literature, the official world is always
being subverted through modes of the rhizome. 1 Mikhail Bakhtin, 1981, The Dialogic Imagination: Four Essays by M. M. Bakhtin, Caryl Emerson,
Michael Holquist (eds.), Austin: University of Texas Press, p. 668.
2 Mockery, abuse, and embarrassment in the marketplace seem to be a European phenomena. By
comparison, in Native American cultures like that of the Hopi, laughter itself is often sacred and lacks
cynical undertones, a fact which is confirmed by their rituals in the plaza, the centre of the pueblo.
Here rituals in which clowns allude to the sexual act and the process of defecation are performed without
embarrassment and accompanied by laughter that contains no derision. See Richard Erdoes and Alfonso
Ortiz 1998, American Indian Trickster Tales, London: Penguin, p. xxi.
3 Peter Stallybrass and Allon White, 1986, The Politics and Poetics of Transgression, Ithaca: Cornell
University Press, p. 36. Rushdie We encounter these two realms,
Deleuze’s arborescence versus rhizomatics, in authors like Grass, Irving, Rushdie,
Tournier, Garcia Marquez, but also in Bruce Chatwin’s Songlines, to add an
example from Australia, where the rhizomatic world of the indigenous peoples
clashes with Western racism and its arborescence. As jazz and blues are part of
the rhizomatic world giving tragicomic hope to African Americans so are the
songlines, that intricate web of dreaming paths as the manifestation of an ancient
nomadic culture. The trickster and the picaro are the great wanderers and
blasphemers of the mythological and fictional world challenging the Gods, the
rulers, and the sedentary bourgeois who hold still in the face of misrule. A
question that remains is: Who will write the great heteroglot novel that parodies
the Bush era and America’s foreign policy? To date it has not yet been produced. 4 Stallybrass and White, p. 42. 5 Jack Zipes (ed.), 1987, The Complete Fairy Tales of the Brothers Grimm, New York: Bantam, p. 659. f
19 Ibid., p. 98. Endnotes ack Zipes (ed.), 1987, The Complete Fairy Tales of the Brothers Grimm, New York: Bantam, p. 659. 78 6 Günter Lewy, 2000, (1964), Nazi Germany and the Catholic Church, New York: Da Capo Press, p. 292. 6 Günter Lewy, 2000, (1964), Nazi Germany and the Catholic Church, New York: Da Capo Press, p. 292. 7 Ibid., p. 279, for example: ‘the veteran National Socialist priest Father Senn…in 1934 hailed Hitler
as the tool of God, called upon to overcome Judaism’. 7 Ibid., p. 279, for example: ‘the veteran National Socialist priest Father Senn…in 1934 hailed Hitler
as the tool of God, called upon to overcome Judaism’. 8 Ibid., p. 279. 9 Mikhail Bakhtin, 1984, Rabelais and his World, Bloomington: Indiana University Press, pp. 192-93. 10 J
d i P
l R di
1972 Th T i k
A S
d i A
i
I di
M h l
N
Y k 9 Mikhail Bakhtin, 1984, Rabelais and his World, Bloomington: Indiana University Press, pp. 192-93. Mikhail Bakhtin, 1984, Rabelais and his World, Bloomington: Indiana University Press, pp. 192-93. 10 Jung quoted in Paul Radin, 1972, The Trickster: A Study in American Indian Mythology, New York:
Schocken Books, p. 198. p
11 Lewis Hyde, 1998, Trickster Makes this World, New York: North Point Press, p. 90. y
12 Hannah Arendt, 1973, The Origins of Totalitarianism, New York: Harcourt Brace & Company, p. 35 Arendt, 1973, The Origins of Totalitarianism, New Yo 13 Mikhail Bulgakov, 1996, The Master and Margarita, New York: Random House, p. 63. 14 For example, the work of Patricia Merivale, 1990, ‘Saleem Fathered by Oskar: Intertextual Strategies
in ‘Midnight’s Children’ and ‘The Tin Drum’, Ariel: A Review of International English Literature, vol. 21, no. 3, pp. 5-21; Rudolf Bader, 1984, ‘Indian Tin Drum’, The International Fiction Review, vol. 11,
no. 2, pp. 75-83; and E. W. Herd, 1989, ‘Tin Drum and Snake-Charmer’s Flute: Salman Rushdie’s Debt
to Günter Grass’, New Comparison, vol. 6, pp. 205-18. 15 K.M. Ashley, 1990, Victor Turner and the Construction of Cultural Criticism, Bloomington: Indiana
University Press, p. xviii. 16 Salman Rushdie, 1980, Midnight’s Children, New York: Knopf, p. 294. 17 John Clement Ball, 1998, ‘Pessoptimism: Satire and the Menippean Grotesque in Rushdie’s Midnight’s
Children’, English Studies in Canada, vol. 24, no. 1, pp. 61-81, at 73-4. 17 John Clement Ball, 1998, ‘Pessoptimism: Satire and the Menippean Gro
Children’, English Studies in Canada, vol. Endnotes 24, no. 1, pp. 61-81, at 73-4. 18 David Price, 1994, ‘Salman Rushdie’s Use and Abuse of History in Midnight’s Children’, Ariel: A
Review of International English Literature, vol. 25, no. 2, pp. 91-107, at 96-106. 19 Review of International English Literature, vol. 25, no. 2, pp. 91-107, at 96-106. 79 79 6
Les fees ont soif: Feminist, iconoclastic
or blasphemous?
Maria-Suzette Fernandes-Dias In his 1993 book, Blasphemy, David Lawton is of the opinion that blasphemy in
the arts is healthy because it ‘often registers the irruption of a new reading
community’1 and marks such a community’s rite of passage. Women’s writing
has been an important site of blasphemy in the twentieth century. Blasphemy
has provided a formerly marginalised group with a medium though which to
assert its rights against an existing social and cultural order that abhors
transformation and resists it. It does this by wielding the power of religious
constructs about the sacrosanctity of dogmas and beliefs, the naturalness of civil
codes of ethical and moral propriety, and the necessity of judicial provisions
like repressive censorship and blasphemy laws. This chapter examines a literary
scandal that marked Quebec’s rupture from traditionalism and religious
institutionalism, and the emergence of its post-Quiet Revolution feminine identity. The scandal of Les fees ont soif erupted in 1978 — more than a decade after
the Quiet Revolution (1960–66) and Quebec’s rejection of traditionalist spiritual
values, classical education, church-controlled and inspired social welfare
institutions and its rural, agricultural heritage, and its assertion of a new
Quebecois identity. Feminine emancipation was still in its embryonic stage. The
Catholic Church, despite its waning political clout, exercised nevertheless,
considerable influence on the behaviour of women.2 A decade of secular outlook
had not been long enough to dismantle traditional constructs about the role of
women and centuries of cultural and social conditioning about male superiority,
or to undo the imposing omnipresence of retrograde patriarchal ideologies in
newly established secular institutions. Frustration, coupled with the emergence
of a second wave of feminism in Western Europe and the USA, catalysed the
surfacing of an almost militant and utopian feminist movement in Quebec. In
1976, Quebec witnessed the emergence of L’Autre Parole, a publication by women
who identified themselves as Catholics and feminists and asked for the
de-gendering of religious practices and discourses to include the presence and
the voice of women. Louky Bersianik’s l’Eugelionne, published the same year,
parodies sacred male writings, especially the Bible and the writings and theories
of Freud and Lacan, satirically denoted as Saint Siegfried and 81 Negotiating the Sacred II Saint-Jacques-Linquant. L’Eugelionne challenges, deflates, and deconstructs
androcentric ideology, its sexist discourse and practices, its misogynist reality,
and its harmful dichotomies. 6
Les fees ont soif: Feminist, iconoclastic
or blasphemous?
Maria-Suzette Fernandes-Dias Owing to its representational power to encompass physical and sexual
difference within the existing patriarchal discourse and to deconstruct negative
myths and images, theatre provided feminist collectives with a transgressive
and performative space in which to pursue their quest for a female identity. Well known examples of this feminist iconoclastic fervour are plays like Si
Cendrillon pouvait mourir (If Cindrella could die) (1975) and La Nef des sorcières
(A Clash of Symbols) (1976), which directly attack stereotype female role models. Quebec decries blasphemy On 16 May, 1978, the Arts Council of Montreal informed the director of the
Théâtre du Nouveau-Monde, Jean-Louis Roux, about their decision not to fund
Les fees ont soif. The president of the council publicly called this play ‘a piece
of shit’, ‘trash’ and challenged local newspapers to publish three pages of its
script. The day after this declaration, La Presse published an extract of the play
and by the end of the month, the editor of Devoir, Michel Roy, called for a censor. By June, the debate over the play mobilised public opinion, igniting another
Querelle de Tartuffe.3 While rehearsals continued, organisations such as the
Association of Theatre Directors, the Human Rights League, Quebecois Writers’
Union, and the International Institute of Theatre filed petitions against a potential
censor. The only female member of the Council, Mme Thérèse Lamarche resigned
in protest against the council’s decision. On 11 November, when the play was
advertised to be staged, the battle seemed won. However, the crusade had just
begun. The play premiered on 25 November amidst protests by the extreme
right, with reparation vigils organised by the Archdiocese of Montreal and
congregations even picketing the theatre en masse to recite the rosary while the
play was being performed. Three days later, the Archbishop of Quebec, Mgr. Paul Grégoire, denounced the play for its vulgarity and its frivolous portrayal
of the Holy Virgin as ‘a puppet, an invention of male domination, a figment
responsible for the alienation of women’.4 A judicial imbroglio ensued. Following a plea lodged by Emile Colas on behalf
of Young Canadians for Christ, Quebecois Catholic Parents’ Association, Farmers’
Association and the Quebec State Council of the Knights of Columbus, the printed
version of the play was banned by the High Court. In January 1979, the decision
was reversed on the basis of a technicality and a petition signed by intellectuals,
including Simone de Beauvoir, Julia Kristeva, Phillippe Sollers, Denis Roche and
Christiane Rochefort. Colas, appealed to the Supreme Court and lost. More judicial
proceedings followed until the Supreme Court declared, in 1980, that it would
not hear any more cases against the work, stating that ‘any injunction against
a work of art having considerable social impact, should come from the Public 82 Les fees ont soif: Feminist, iconoclastic or blasphemous? Quebec decries blasphemy Prosecutor of the province, given that the work affects the whole society, not
isolated individuals’.5 Since 1980, Les fees ont soif has been successfully staged several times —
even during the papal visit to Canada in 1984 — and as recently as July 2005
during the Festival de Fringe in Montreal, without being decried as blasphemous
or scandalous. In the literary and in the cultural paradigm, Les fees ont soif is
considered as a prominent marker of the post Quiet Revolution assertion of the
feminine identity and the social rupture from religious dogmatism in Québec. Why? Les fees ont soif: feminist iconoclasm and blasphemy Boucher sought to explore not the myth of the Virgin but the wife and mother
repressed in her, as she explains in the foreword of her play: Since childhood, the image of the Holy Virgin walked in my body and
my head. The woman hidden in her haunted me. Where would it be
possible to meet her?…An entire bachelor culture had projected and
transferred its fantasies of virginity on the mother of Jesus and all other
women. An androcentric culture had only fabricated a single archetype
of reference for women — that of the virgin. A woman who does not
enjoy sensual pleasures, be it a mother or a whore. Women are thus
exiled from the gratification of their bodies.8 Since childhood, the image of the Holy Virgin walked in my body and
my head. The woman hidden in her haunted me. Where would it be
possible to meet her?…An entire bachelor culture had projected and
transferred its fantasies of virginity on the mother of Jesus and all other
women. An androcentric culture had only fabricated a single archetype
of reference for women — that of the virgin. A woman who does not
enjoy sensual pleasures, be it a mother or a whore. Women are thus
exiled from the gratification of their bodies.8 A proponent of the same tradition as Kazantzakis, Scorsese and Gibson,
Boucher tried to emancipate the Virgin Mary from the conflict between the
contradictory elements of the human and the divine that co-exist as part of her
personality. Adhering to the iconoclastic conviction that the Divine-Human
beings remain essentially divine by overcoming their human impulses and urges
and by seeking the ‘higher mission’ through constant prayer, Boucher induces
her character of the Virgin Mary to expunge the divine and embrace the human
by rejecting prayer and abnegation from her life: ‘Once upon a time, one
day…and that is today, I began to distort the Angelus’.9 ‘To make a statement,
one must come down from the cross’.10 In the theatrical production, Boucher divided the stage into four distinct
areas, offering a representational space to each archetype — Marie, the mother
in a kitchen; Madeleine, the whore in a bedroom; and the Virgin Mary as a statue
raised on a pedestal towards the back of the stage. Les fees ont soif: feminist iconoclasm and blasphemy In his book, Sorcières (1862), French historian Jules Michelet describes fairies as
the proud and fantastical queens of Gaul, who brazenly turned their backs on
Christ and his apostles and continued to dance. For this impudence, they were
imprisoned in containers that would be opened only at the end of time. Drawing
on this metaphor in Les fees ont soif (translated by Alan Brown as The Fairies are
Thirsty), playwright, Denise Boucher, attempted to deconstruct the role played
by myth, image and language in the formation of women’s socio-cultural identity
by creating an iconoclastic feminine trilogy of the Virgin Mary, the Mother and
the Whore as a satirical counterpart to the Holy Trinity of the Father, the Son
and the Holy Spirit, to depict how patriarchal tradition has incarcerated women
in stereotypical roles of submission. The three characters rebel against their
archetypal roles and invite the audience to ‘imagine…imagine…imagine’ a
different world, recreated by women. In Mythologies, Roland Barthes elaborates how societies create myths to
legitimise power structures.6 The rapport between power and myth being
symbiotic in nature, power creates and legitimises the myth and the myth
perpetuates itself while ensuring the official recognition of power. Thus, a myth
is, according to Barthes, a social construct which presents itself as natural, but
is in fact, fabricated by History through social and cultural conditioning. More than just a system of theatrical representation, the archetypes of the
Virgin, the Mother and the Whore constitute the foundations of an androcentric
ideology that has strived to alienate the woman from her own history and her
body. The originality of Boucher’s play was to create with these archetypes, a
functioning trinity capable of denouncing its fears, its deprivations and its stasis. The Virgin, despite her status as a female religious icon, is more of a feminine
model than a feminine creation. The patriarchy accords her power and authority,
but only to serve as the model of the perfect, submissive woman. As Julia Kristeva
illustrates in her article ‘Stabat Mater’,7 the religious and secular representation
of femininity in our civilisation is characterised by motherhood — the 83 Negotiating the Sacred II desexualised maternal representation of women — a myth reinforced by the
dogma of the Virgin Mother. Les fees ont soif: feminist iconoclasm and blasphemy In the foreground is the
neutral space, a space free from patriarchal influence, where the three characters
discover themselves, express themselves freely, and develop their solidarity. More than the depiction of the Virgin, flanked by her supposed alter egos, Marie
the wife and mother and Madeleine the whore, it was Boucher’s transgression
from the terrain of art — or rather her subversive use of art — to attack the
overpowering influence of the church on the behaviour of women and the
sanctity of the dogmas of the Virgin and the Holy Spirit, that drew the wrath
of the clergy. The statue of the Virgin is portrayed as holding a thick and heavy chain, a
parodied representation of the rosary and making a mockery of the Litany of
Loreto: I am an image I am a portrait I have my feet planted in plaster 84 Les fees ont soif: Feminist, iconoclastic or blasphemous? I am the queen of nothingness
I am the door to an abyss
I am the unconsummated marriage of priests (…)
I am the mirror of injustice
I am the throne of slavery
I am the elusive sacred vessel
I am the obscurity of ignorance (…)
I am the help of the useless
I am the tool of weaknesses
I am the decayed symbol of rotting abnegation
I am more oppressive and more repressing than all words (…)
I am the imagined image
I am she who does not have a body
I am she who never menstruates.11 The eternally idolised Virgin leaves her pedestal to occupy the same space as
the housewife and the whore, to express her doubts and her aspirations. She
agrees with Marie, the mother and wife, about the impossibility of articulating
a feminine voice within constraints of a predominantly masculine discourse. Says Marie, speaking to her demented mother: Says Marie, speaking to her demented mother: You who suffered for your subservience, why are you trying to force
me into submission as well? (…) you liked the parish priests. They turned
you away from your body. From your man. And from me. They robbed
you of yourself. (…) They cheated you, mama. Their discourse does not
belong to us. It does not possess the vocabulary to express what I feel,
what I am looking for. Les fees ont soif: feminist iconoclasm and blasphemy It denies me my identity.12 Stepping into the neutral space, the Statue of the Holy Virgin continues: Stepping into the neutral space, the Statue of the Holy Virgin continues: They profess: ‘Silence is golden’ in order to crush under their feet, the
silent female herds. I too have to shut up so that I can listen to Him
always. I owe it to Him to sport the Buddha’s grin, the Sphinx’s head,
the Virgin’s eye.13 Pushing the limits of decorum and of what was considered as representable in
a theatrical production, Boucher not only engages in vulgarising discussion
about sexuality, incest and rape, but even involves the Holy Virgin in doing so. Like a frolicsome teenager, the Statue keeps singing repeatedly: ‘Someday my
prince will come for me’.14 As an almost shocking sequel to Madeleine the whore’s drunken monologue
about her depression and her growing aversion to her profession, the Statue
moves to the neutral space, chanting a commercial for tampons: 85 Negotiating the Sacred II On those dreary day, lady, thanks to Tantax, feel free! Go horse riding,
play tennis, swim! Tantax is discreet. Tantax protects you. Tantax allows
you the freedom of movement! Use Tantax! Be modern! Be free! Be
Tantax!15 The Statue incites the women, Marie and Madeleine, to articulate what was than
considered ‘unspeakable’ and joins them in voicing in a nursery rhyme, their
womanly fears of solitude, ageing and independence in a male dominated society: Statue: Fear
Marie: Fear of going mad. Madeleine: Fear of being alone. Marie: Fear of growing ugly. Madeleine: Fear of becoming too fat. Statue: Fear of knowing too much. Marie: Fear of being touched. Madeleine: Fear of laughing too much. Marie: Fear of weeping. Statue: Fear of speaking out. Marie: Fear of making a fool of myself. Madeleine: Fear of being a hussy. Marie: Fear of becoming frigid. Madeleine: Fear of an orgasm. Marie: Fear of not having an orgasm. Madeleine: Fear of being free. Marie: Fear of my husband. Statue: Fear of mice. Statue: Fear.16 Statue: Fear Marie: Fear of going mad. Madeleine: Fear of being alone. Madeleine: Fear of becoming too fat. Statue: Fear of knowing too much. Madeleine: Fear of laughing too much. Marie: Fear of weeping. Statue: Fear of speaking out. Marie: Fear of making a fool of myself. Madeleine: Fear of being a hussy. Marie: Fear of becoming frigid. Madeleine: Fear of an orgasm. Les fees ont soif: feminist iconoclasm and blasphemy Marie: Fear of not having an orgasm. To reinforce her metaphorical accusation against a system that legitimises the
use of abuse as a means of controlling women, Boucher had the actress playing
the role of the Holy Virgin, also playing the role of the rapist of the whore and
justifying: ‘Raping a prostitute does not amount to rape’.17 In an almost ironic theatrical twist, the Virgin and Marie take the centre stage
to condemn the judiciary and lament the judicial fiasco that ensued as a result
of the rapist’s trial: Marie: The trial has all the elements of a masquerade. All the humiliation,
all the misery of a dispossessed woman. Statue: The judge came across as objective. The lawyers as well. None
of them felt implicated. (…) None of them recognized in the victim the
image of their mother, their daughter or their spouse. Patrimony remained
unaffected. As if to say that our heritage permits the rape of women. 86 Les fees ont soif: Feminist, iconoclastic or blasphemous? During the trial, the question that aroused the most interest and the
anxiety and made us forget even the accused, the question that became
the deciding one for the acquittal of the accused was: ‘Did the victim
resist?’18 During the trial, the question that aroused the most interest and the
anxiety and made us forget even the accused, the question that became
the deciding one for the acquittal of the accused was: ‘Did the victim
resist?’18 Some of the lines articulated by the Virgin in this scene are tainted with crude
and scandalous sous-entendus about sexuality; for instance, the statue of the
Virgin says: ‘No matter where. No matter when. No matter how. A tail can thrill. Everyone knows that.’19 Some of the lines articulated by the Virgin in this scene are tainted with crude
and scandalous sous-entendus about sexuality; for instance, the statue of the
Virgin says: ‘No matter where. No matter when. No matter how. A tail can thrill. Defence against allegations of blasphemy Elaborating on the characteristics of blasphemy, Lawton points out that those
who have set out deliberately to blaspheme, ‘set out to argue a system of belief
or to commend a way of life, and what is taken to be blasphemy is often, in their
view, incidental to their aims or a misrepresentation of them. They tend to feel
and say that their words have been taken out of context’.25 The creators of the Les fées ont soif always avowed what Bakhtin would call,
their ‘carnivalesque’ aspiration to undermine and destroy the hegemony of
Quebecois paternalistic ideology and to elucidate feminine potentials as an
alternate conceptualisation of reality. The music composer, Jean-François
Garneau, fearlessly affirmed: ‘Our entire culture is a culture of celibate men who
have rejected women and children through the ages.’26 Comedians Michèle
Magny and Sophie Clement, who played the roles of the Mother and the Whore
respectively, publicly asked: ‘Up to now, why did our mothers maintain their
silence on topics like rape, incest, prostitution and their own self privation of
pleasure?’27 As for the use of coarse language, Boucher deemed it necessary
because stereotypes are not easily shattered. While the Church claimed its ‘right to respect’ and affirmed its dedication
to the emancipation of women, editor of Le Devoir, Michel Roy, expressed the
opinion that Boucher could have achieved her objective on the dramatic, lyrical
and ideological planes by withdrawing the passages that many believers
considered as blasphemous. The issue of blasphemy normally arises in a community that is divided, and
it generally arises because the community is divided. For the readership and the
viewers of Les fees ont soif, the boundaries between theatrical representation
and reality began to blur. Even for Fr. Emile Legault, founder of the Companions
of Saint-Laurent and a strong defender of theatre in Quebec, in a daring television
interview praised the play as a ‘marvellous theological intuition’ but made the
following observation: Five minutes through the play, I began to say to myself that the author
was wrong in not presenting to us her characters as clients of a
psychiatric clinic. Les fees ont soif: feminist iconoclasm and blasphemy Everyone knows that.’19 In Luke 12:10, blasphemy against the Holy Spirit is decreed as unforgivable,
but Boucher did not consider it as a stumbling block to her feminist iconoclasm
20 — she makes the Virgin call holy prophets ‘eunuchs in flesh and mind’,20
deplore her role in the story of creation as the sacrificing mother, call the Holy
Spirit a bird, and denote herself as the ‘Immaculate of all male obsessions’ and
the ‘queen of the mute’: I was given a bird as a husband. My son was robbed from me through
all the ages. He was given a celibate, jealous and eternal father. I was
sculpted in marble and made to crush the serpent with all my strength.21 I was given a bird as a husband. My son was robbed from me through
all the ages. He was given a celibate, jealous and eternal father. I was
sculpted in marble and made to crush the serpent with all my strength.21 Questioning the dogma of the Immaculate Conception is interrogating the dogma
of original sin and the virgin birth and amounts to attempting to dismantle the
very foundation of the Christian monotheist civilisation. As Leonard Levy
demonstrates in Blasphemy, ‘The virgin birth was always a likely subject for
blasphemous ridicule from Taylor’s mad utterances in the seventeenth century,
to Paine’s sarcasms, to Yeat’s “Stick of Incense”’.22 In the denouement of Les fees ont soif, in which each of the characters rebels
against her culturally imposed social role, the Virgin breaks her plaster cast,
drops her rosary and releases the serpent, the symbol of sexual desire, a creature
cursed by God in the story of Creation, and expresses her disgust of the existing
order: I can’t take it anymore. I won’t take it anymore. No. No. I don’t want
this sarcophagus. I don’t wish to be worshipped as a statue whilst I am
being denigrated, despised and demeaned in every woman. I no longer
want to be an alibi for this cursed race of old boys!23 Commenting upon the iconoclasm in the play, feminist Claire Lejeune who
prefaced the published version of Les Fées ont soif, describes the play as ‘a
diabolic Sabbath in which the sense of sin and the sacrosanct distance between
the mother and the whore are simultaneously lost in public’.24 87 87 Negotiating the Sacred II Defence against allegations of blasphemy If she had, then the Madonna would not have been
the Holy Virgin in person but a mentally disturbed person who thinks
she is the Holy Virgin.28 Dailies like La Presse reported the angry response from male protesters that
the personalities portrayed in the play were a far cry of what their mothers or
their sisters were. As for those who practised the oldest profession on earth,
they did so ‘out of their own free will because it paid fifty dollars a quarter of
an hour, better paid than most factories’.29 88 Les fees ont soif: Feminist, iconoclastic or blasphemous? And then, of course, there were rejoinders from agnostics, who felt it was
time to re-examine the dogma of ‘virginity of a mother’,30 and frenzied feminists
from the Council for the Status of Women, who accused Quebecoise society of
muzzling women and artists radically. Detractors representing this extreme,
such as Micheline Carrier, retorted: ‘There has never been an occasion for women
to interpellate the Church without being reminded to respect the established
order, because the Church refuses to engage in a dialogue with women as if to
say that the Holy Spirit breathed only upon the male half of humanity.’31 A study of the religious history of Quebec indicates that the blasphemy law
in Quebec, formulated in 1806, is very precise — the most severe form of
punishment was, undoubtedly, cutting the tongue of the offender. Drawing on
this analogy, Nancy Huston articulates the conclusion of her article on the
reception of Boucher’s play: In North America, almost twenty centuries after the birth of Christ, the
belief still prevails that — just as the best way to keep a fish from
smelling is to cut off the nose — the best way to keep a fairy from being
thirsty is to cut out her tongue.32 From outrage to acceptance The Quebecois society of the 1970s strove to reject Les Fées ont soif from its
cultural discourse for its vulgar language and its treatment of then proscribed
themes such as menstruation, prostitution, violence against women and women’s
rage and madness; from the social discourse for its religiously blasphemous
import and its virulent onslaught on taboos and myths profoundly anchored in
the history of the people; and from the artistic discourse for its amateurish
melange of genres like poetry, theatre and opera. However, as is often the case
with censorship and succès de scandale, despite its modest artistic quality and
its espousal of what now seems to be a utopian cause of the feminism of
difference, the play has found a niche in the literary history of Quebec as a
daring attempt to create social justice for women by calling for a profound
restructuring of society and of the way in which people think and experience
the world. By stressing that ‘the personal is political’, breaking the silence over
issues like rape and incest, and, of course, rendering the discourse with much
potence by attributing it to the character of the Holy Virgin, this play made the
social inequality of women a public and not merely a private problem. How does one explain the public’s nonchalance in the consequent years
leading to the present, to the staging of what was once disparaged as
blasphemous? Envisioning the humanity of the divine and evoking or expressing
it in art is still considered as an attack on conventional piety as we have seen in
the public’s reaction to Scorsese’s The Last Temptation of Christ (1988) or Mel
Gibson’s The Passion of Christ (2004). It is therefore understandable how a 89 Negotiating the Sacred II recognisably human dimension and an avant-garde feminist depiction of the
Virgin as a woman compelled to suppress her corporeal desires, a mother who
resents having to sacrifice and grieve for her son in silence in accordance to a
higher plan (‘I went through this excessive agony of making a child see the light
of the day and the darkness of the unknown higher plan. Since I gave him life,
I became in many ways responsible for his death as well’33 ), and as a rebel who
finally breaks away from her role, challenged the beliefs and expectations of the
Quebecoise Catholic community. David Lawton, Blasphemy, 1993, Philadelphia: University of Pennsylvania Press, p. 139.
2 In Les religieuses sont-elles féministes?, Micheline Dumont speaks about the increase in the number
of Quebecoise nuns (from 1850 to 1965, an increase from 650 to 43,274) and explains that taking orders
was a form of individual emancipation, that the Church provided women who became nuns with an
access to life which the Quebecoise society would have otherwise denied them. Micheline Dumont,
1995, Les religieuses sont-elles féministes? Montreal: Bellarmin. From outrage to acceptance But with the passage of time, a community that was already divided and in
a state of transition gradually lost the importance it accorded to religion as a
factor of social cohesiveness. Therefore, an artistic creation that was once
considered as morally and spiritually objectionable and had overstepped the
metaphorical and ideological limits imposed by the traditional order, did not
shock the Quebecois society anymore. The focus of the public has thus shifted
from the alleged blasphemous content to the humour and the social message
embodied in the play, which Ingrid Pux who played the role of the Virgin in
1995 when the troupe Majeure presented the play, describes as being ‘topical
because it shows women fossilised in stereotypes’.34 Far from being an
indictment against men, as it was viewed when it premiered amidst the
re-emergence of feminism as ‘feminism of difference’, for the audience today the
play, ‘transmits the enduring message that the world can only change when
women and men work together to bring about change instead of imprisoning
the other in proverbial clichés’.35 I must confess, however, the moral conflict that assailed me while writing
this chapter. Boucher’s depiction of the Virgin scandalised me, for, as a Catholic,
I have always revered the Holy Virgin for her emancipatory role in the history
of redemption of Mankind. As a daughter, a wife and a mother, I look up to her
not as a servile instrument in the fulfilment of a greater plan but as a role model
exemplifying dedication, fortitude and courage — she is one of the reasons why
I feel privileged to be a woman. Despite my reservations and my personal
religious convictions, as a Bakhtinian, I laud Les fées ont soif for its artistic
iconoclasm, which induced society to reconsider and re-evaluate its attitude
towards women; and for its heteroglossic capacity to reflect a multitude of
dissenting voices in a dialogic relationship, interrogating the predominance of
a single repressive ideological discourse. 1 David Lawton, Blasphemy, 1993, Philadelphia: University of Pennsylvania Press, p. 139. 34 As reported by Renée Larochelle in her preview, ‘“Théâtre. Les fées ont soif”. Au fil des Événements”,
Université Laval’, dated 12 October 1995 of the performance to be staged on 13 and 14 October, 1995,
at Café theatre des Fourberies. http://www.scom.ulaval.ca/Au.fil.des.evenements/1995/45/011.html
(Viewed 26 August 2008.) 3 Querelle de Tartuffe (1693-94). As evidence that the clergy in the New World could be as controlling
as that of the Old, the Church banned the proposed production of Molière’s Le Tartuffe. Even 30 years
after its creation, the play was still considered controversial in France. In the New World, the controversy
was more of a battle of power between the state and the clergy than a battle over morality. 7 Julia Kristeva, 1985, ‘Stabat Mater’, Poetics Today vol. 6, no. 1-2, pp. 133-152.
8 Les fees ont soif: Feminist, iconoclastic or blasphemous? 3 Querelle de Tartuffe (1693-94). As evidence that the clergy in the New World could be as controlling
as that of the Old, the Church banned the proposed production of Molière’s Le Tartuffe. Even 30 years
after its creation, the play was still considered controversial in France. In the New World, the controversy
was more of a battle of power between the state and the clergy than a battle over morality.
4 Lise Gauvin, ‘Introduction’, in Denise Boucher, 1989, Les fées ont soif, Montréal: l’Hexagone, p. x.
For a more recent version of the play, see Denise Boucher, Les fées ont soif, Éditions Typo, 2008. This
play has also been translated in English by Alan Brown as The Fairies are Thirsty, Talonbooks, 2008.
This translation can also be found in the Anthology of Québec Women's Plays in English Translation,
Volume I (1996-1986), (edited by Louise H, Forsyth) Playwrights Canada Press, Toronto, 2006. An
English translation can also be found in Plays by French and Francophone Women: A Critical Anthology 3 Querelle de Tartuffe (1693-94). As evidence that the clergy in the New World could be as controlling
as that of the Old, the Church banned the proposed production of Molière’s Le Tartuffe. Even 30 years
after its creation, the play was still considered controversial in France. In the New World, the controversy
was more of a battle of power between the state and the clergy than a battle over morality.
4 Lise Gauvin, ‘Introduction’, in Denise Boucher, 1989, Les fées ont soif, Montréal: l’Hexagone, p. x.
For a more recent version of the play, see Denise Boucher, Les fées ont soif, Éditions Typo, 2008. This
play has also been translated in English by Alan Brown as The Fairies are Thirsty, Talonbooks, 2008.
This translation can also be found in the Anthology of Québec Women's Plays in English Translation,
Volume I (1996-1986), (edited by Louise H, Forsyth) Playwrights Canada Press, Toronto, 2006. An
English translation can also be found in Plays by French and Francophone Women: A Critical Anthology
(edited and translated by Christiane P. Makward and Judith G. Miller with an annotated bibliography
by Cynthia Running-Johnson) The University of Michigan Press, 1994.
5 Ibid., p. xi.
6 Roland Barthes, 1957, Mythologies, Paris: Seuil.
7 Julia Kristeva 1985 ‘Stabat Mater’ Poetics Today vol 6 no 1 2 pp 133 152 Endnotes 90 Les fees ont soif: Feminist, iconoclastic or blasphemous? 3 Querelle de Tartuffe (1693-94). As evidence that the clergy in the New World could be as controlling
as that of the Old, the Church banned the proposed production of Molière’s Le Tartuffe. Even 30 years
after its creation, the play was still considered controversial in France. In the New World, the controversy
was more of a battle of power between the state and the clergy than a battle over morality. 4 Lise Gauvin, ‘Introduction’, in Denise Boucher, 1989, Les fées ont soif, Montréal: l’Hexagone, p. x. For a more recent version of the play, see Denise Boucher, Les fées ont soif, Éditions Typo, 2008. This
play has also been translated in English by Alan Brown as The Fairies are Thirsty, Talonbooks, 2008. This translation can also be found in the Anthology of Québec Women's Plays in English Translation,
Volume I (1996-1986), (edited by Louise H, Forsyth) Playwrights Canada Press, Toronto, 2006. An
English translation can also be found in Plays by French and Francophone Women: A Critical Anthology
(edited and translated by Christiane P. Makward and Judith G. Miller with an annotated bibliography
by Cynthia Running-Johnson) The University of Michigan Press, 1994. 5 6 Roland Barthes, 1957, Mythologies, Paris: Seuil. 7 Julia Kristeva, 1985, ‘Stabat Mater’, Poetics Today vol. 6, no. 1-2, pp. 133-152 8 Boucher, op. cit., p. 39. 9 Ibid., p. 46. p
10 Ibid., p. 52. p
11 Ibid., p. 60. p
11 Ibid., p. 60. p
12 Ibid., p. 77. p
12 Ibid., p. 77. p
13 Ibid., p. 85. p
14 Ibid., pp. 47, 48. pp
15 Ibid., p. 53. p
16 Ibid., p. 53. p
17 Ibid., p. 92. p
18 Ibid., p. 93. 19 Ibid., p. 93. p
20 Ibid., p. 77. p
21 Ibid., p. 50. 22 Leonard Levy, 1993, Blasphemy, New York: Alfred Knopf, p. 534. 23 23 Boucher, op. cit., p. 95. 23 Boucher, op. cit., p. 95. p
p
24 Levy, op. cit., p. 28. 25 David Lawton, 1993, Blasphemy, Philadelphia: University of Pennsylvania Press, p. 2. 26 26 Jean-François Garneau, 1978, Texte liminaire de l’édition originale, Intermède quoted by Lise Gauvin,
‘Introduction’, in Denise Boucher, 1989, Les fées ont soif, Montréal, l’Hexagone, p.10. 27 27 Garneau, op. cit. 27 Garneau, op. cit. 35 Larochelle, op. cit. 6 Roland Barthes, 1957, Mythologies, Paris: Seuil. 7
The body of Christ: Blasphemy as a
necessary transgression?
Carolyn D’Cruz and Glenn D’Cruz This chapter was originally prefaced with an audio-visual presentation showing key
scenes from Martin Scorsese’s film, The Last Temptation of Christ, cut to the folk
hymn, ‘Go Tell Everyone’, which is reproduced below. The video segued into a short
duologue, which dramatised the ambiguity inherent in God’s call. The presentation
concluded with the same hymn accompanied by images of the poor, the marginalised
and the powerful. God’s spirit is in my heart
He has called me and set me apart
This is what I have to do, what I have to do
He sent me to give the good news to the poor
Tell prisoners that they are prisoners no more
Tell blind people that they can see
And set the downtrodden free
And go tell everyone, the news that the kingdom of God has come
And go tell everyone, the news that God’s kingdom has come.1 God’s spirit is in my heart
He has called me and set me apart
This is what I have to do, what I have to do
He sent me to give the good news to the poor
Tell prisoners that they are prisoners no more
Tell blind people that they can see
And set the downtrodden free
And go tell everyone, the news that the kingdom of God has come
And go tell everyone, the news that God’s kingdom has come.1 Did you hear the call? What call? I think it’s the call from God. You think? If it is the voice of God shouldn’t you know? But it could be the voice of the devil fooling me to believe it is God. How will I
ever know for certain? You won’t. You think? If it is the voice of God shouldn’t you know? But it could be the voice of the devil fooling me to believe it is God. How will I But it could be the voice of the devil fooling me to believe it is God. How will I
ever know for certain? You won’t. But it could be the voice of the devil fooling me to believe it is God. How will I
ever know for certain? In the Last Temptation of Christ, Martin Scorsese’s Jesus heard voices. Endnotes 28 L
l
d i Ed
d R bill d 1978 ‘F
d
l
’ L D
i
18 D
b
(
i li
) 28 Legault, quoted in Edmond Robillard, 1978, ‘Forum des lecteurs’ Le Devoir, 18 December (my italics). 28 Legault, quoted in Edmond Robillard, 1978, ‘Forum des lecteurs’ Le Devoir, 18 December (my italics). 29 E. Robillard, 1978, ‘Opinions’, La Presse, 15 December. Legault, quoted in Edmond Robillard, 1978, Forum des lecteurs Le Devoir, 18 December (my italics). 29 E. Robillard, 1978, ‘Opinions’, La Presse, 15 December. 29 E. Robillard, 1978, ‘Opinions’, La Presse, 15 December. 30 Bernard Larivière Saint-Colomban, 1978, ‘Arts et Spectacles – Lectures’, La Presse, 15 December. 31 31 Micheline Carrier, 1978, ‘Forum des lecteurs’, Le Devoir, 28 December. 32 Nancy Huston, 1981, ‘Blasphemy in “Nouvelle France” Yesterday and Today’, Maledicta, vol. 5, no. 2, pp. 163-169. 33 Boucher, op. cit., p. 83. 34 As reported by Renée Larochelle in her preview, ‘“Théâtre. Les fées ont soif”. Au fil des Événements”,
Université Laval’, dated 12 October 1995 of the performance to be staged on 13 and 14 October, 1995,
at Café theatre des Fourberies. http://www.scom.ulaval.ca/Au.fil.des.evenements/1995/45/011.html
(Viewed 26 August 2008.)
35 (Viewed 26 August 2008.) 35 Larochelle, op. cit. 35 Larochelle, op. cit. 91 91 7
The body of Christ: Blasphemy as a
necessary transgression?
Carolyn D’Cruz and Glenn D’Cruz He was
not always sure if it was the voice of God calling him, setting him apart to redeem
the world from sin, whether it was the work of the devil leading him into
temptation, or whether it was just his own basic corporeal voice of desire. In
any case, Scorsese’s Jesus struggled between the pull of a divine plan on the
one hand, and temptations of the flesh as part of living a typical life on earth on
the other. In contrast to the many filmic images of Jesus that episodically rise
from the dead on our television screens at Easter time — images embodied by 93 Negotiating the Sacred II the likes Max Von Sydow, Jefferey Hunter and Robert Powell — Scorsese’s
image of Jesus, as he puts it, does not ‘glow in the dark’. Scorsese wanted to
present the world with a Jesus whose human side was not effaced by his status
as a deity. Adapting his filmic text from Nikos Kazantzakis’s novel of the same
name, Scorsese underscores the struggle that Jesus experiences as a man coming
to terms with the temptations of ‘ordinary’ life. He experiences these temptations
as strongly as the voices that call him to a duty not to himself but to others. It is possible to read The Last Temptation of Christ as a text that responds to
Scorsese’s own calling. When Paramount Pictures initially agreed to proceed
with the film in 1983, the studio executives asked Scorsese why he wanted to
make the film; he replied: ‘So I can get to know Jesus better.’2 This would come
as no surprise to Scorsese scholars, as one can hardly miss the Catholic themes
and iconography in films such as Mean Streets and Raging Bull. Indeed, many
academic commentaries on The Last Temptation of Christ note similarities between
this film and its predecessors, arguing that the entire Scorsese canon dramatises
the struggle between the sacred and the profane, the body and the spirit. As
Rolando Caputo notes: Scorsese has so doggedly and publicly pursued this ‘long treasured
project’ [The Last Temptation] that there is almost a hidden implication
that he would have us believe most of his previous films were mere
sketches for a canvas, of which Last Temptation is the final unveiling ... 7
The body of Christ: Blasphemy as a
necessary transgression?
Carolyn D’Cruz and Glenn D’Cruz In retrospect, and not surprisingly, many of Scorsese’s films are a little
bit of Last Temptation, or as Kay more aptly states it, he is ‘synthesising
all his cinematic Scorsesisms into one film.3 Furthermore, it appears that Scorsese heard his own calling to get to know Jesus
better well before he became a film director. He was brought up Catholic, served
as an altar boy and wanted to become a priest before studying film at New York
University. As a ‘true believer’, so to speak, Scorsese was attracted to
Kazantzakis’s novel because he found the novelist’s representation of Christ
accessible. While recounting an episode in his film when Christ says: ‘Lucifer
is inside me, he’s saying I’m not the son of Mary and Joseph, I am the son of
God, I am God’, Scorsese says: So he thinks it’s the Devil inside him saying this, and he believes he is
the worst sinner in the world. I felt that this was something I could relate
to: this was a Jesus you could sit down with, have dinner or a drink
with.4 If we were to give any weight at all to authorial intentionality, as so many people
do, we would be more than hard pressed to accuse Scorsese of wanting to make
a blasphemous film.5 As far as intentions go, Scorsese recalls a priest telling him
that Kazantzakis’ novel was read in Catholic seminaries in order to provoke 94 The body of Christ: Blasphemy as a necessary transgression? debate and discussion. Scorsese states: ‘this is what I hoped the film would do’.6
Defending Scorsese against charges of blasphemy, Jonathan Rosenbaum remarks:
‘the religious doubts about Scorsese, one should stress, are not doubts about
religion, but on the contrary, doubts which could only exist within a system of
religious belief’.7 But regardless of Scorsese’s authorial intentions, despite his careful framing
of The Last Temptation as a fictional text, he was bound to offend the
self-appointed guardians of Christianity over the way the story of Christ should
be told. It ought to be common knowledge that as a text, The Last Temptation
of Christ will be read in a variety of institutional and discursive circumstances
and contexts that generate different meanings. 7
The body of Christ: Blasphemy as a
necessary transgression?
Carolyn D’Cruz and Glenn D’Cruz The point is that the charges of blasphemy were
made by a tightly organised, relatively powerful group of Christian
fundamentalists — most notably the National Federation for Decency led by
Donald Wildmon in the United States — who were able to disrupt the very
production of the film, years before it even made it to the screen. In charting these attempts to censor the film, we get an idea of just how
influential fundamentalism can be in determining who gets to speak about Christ,
and how Christ must be spoken about in order to speak legitimately. It also
shows who gets to define blasphemy. In his book, The Man the Networks Love to Hate, Donald Wildmon recalls
that when he first heard about Paramount’s plans to fund Scorsese’s Last
Temptation, he orchestrated a nationwide campaign to prevent the film’s
production. He exhorted Christians to express their outrage by bombarding
Paramount with letters, phone calls and postcards of protest. ‘Thankfully’,
Wildmon states, ‘this flood of communication convinced Paramount, which had
already spent $2 million on the Last Temptation project, to cancel its plans’.9
Paramount’s withdrawal from the project generated apprehension throughout
the film industry, and for a short while, it seemed that the film would find no
producer. Five years later, Scorsese secured the backing of Universal, a subsidiary of
the Music Corporation of America (MCA), on the proviso that he would produce,
direct and develop his subsequent projects exclusively for them. The company
gave The Last Temptation ‘a budget less than half of the average Hollywood
film’.10 Universal tried a number of methods to defuse religious controversy,
and went as far as hiring conservative religious opinion makers as consultants
for marketing the project (the consultants later resigned after their calls for
revising two thirds of the script went unheeded). Still controversy could not be
defused. This time, the fundamentalist response was even more extreme, and
widespread. In fact, Christians of various denominations joined the
fundamentalist protest all over the world (some even bombed a Parisian theatre). Needless to say, very few protesters had actually seen the film. Nonetheless, the
scale of the protests against the movie was unprecedented. Wildmon engaged
in a media blitz that saw him appear on network television shows such as Oprah
and Good Morning America. 7
The body of Christ: Blasphemy as a
necessary transgression?
Carolyn D’Cruz and Glenn D’Cruz Cinemagoers, for instance, might
be interested in the film’s aesthetic qualities, seminaries might use the film as a
pedagogical tool, and equally some viewers might find the thought of Jesus
Christ having sex on film titillating. As we will see, Christian fundamentalists
used the film to galvanise their constituency against what it saw as moral
corruption. So, if Scorsese was intent on provoking ‘debate and discussion’ on
his particular religious calling, he could not escape doing so among a cacophony
of other receivers of the call. In this sense, Scorsese’s struggles presented through the non-‘glow in the
dark’ Jesus are not unlike our own. Perhaps each of you has heard a calling. Perhaps it is a religious calling and you are certain it is the voice of God. Or
perhaps, in accord with the seemingly secular context of the modern academy,
you might prefer to think of a calling as the voice of conscience or reason. But
for some people, like us for example, even the voice of conscience and reason
does not adequately capture the ‘spirit’ of the call to which we are responding
to today. Now neither of us, the authors of this chapter, are religious in the strict
sense of the term, although we have extremely divergent relationships to the
heritage of Christianity. Yet the call that we underscore here and now cannot
escape the messianic tone of at least one thread of Christianity’s heritage: the
coming of justice. Our philosophical guides for grappling with this heritage and
call are located in works by Emmanuel Levinas and Jacques Derrida in particular. We will come back to these guides later. For now, let us focus on our aesthetic
guide, Martin Scorsese and his fictional, cinematic re-telling of the story of
Christ, which gives the hypothesis of this chapter its impetus: the contention
that so-called ‘blasphemy’ might be a necessary transgression against the
fundamentalist Christian Right, and as an important consequence, an essential
ingredient for creating a more just society. For the record, we do not believe that the film is actually blasphemous, if we
take blasphemy to mean ‘a contemptuous or profane act or utterance, or writing
concerning God’.8 Christ is tempted by flesh in the film, but he does not succumb
to it. He wonders if he is merely mortal, but eventually accepts his divine calling. 95 Negotiating the Sacred II But this is beside the point. 7
The body of Christ: Blasphemy as a
necessary transgression?
Carolyn D’Cruz and Glenn D’Cruz He sent 250 copies of the script to what he calls
‘influential’ Christian leaders, and recalls that his organisation, the American
Family Association (AFA), recorded a three-minute Last Temptation-related radio spot that was
soon airing on 800 Christian radio stations nationwide. We also hurried
to produce a 30-minute television program attacking the movie. It aired
on more than 50 Christian stations and cable networks. We urged people
to protest by writing and calling MCA/Universal. Our AFA phone number 96 The body of Christ: Blasphemy as a necessary transgression? also appeared so people could call and request a special petition asking
local theatre owners, out of respect for Christians in their community,
not to show the film. But this was only the beginning. In July I wrote
to 170,000 Christian pastors and asked them to promote the protest effort
in their churches. I also sent out more than three million letters to
Christian lay people.11 Following the example set by Wildmon and his organisation (by now re-named
the American Family Organisation), other fundamentalist organisations such as
the Campus Crusade for Christ, under the direction of Reverend Bill Bright,
‘offered followers a ‘Last Temptation Battle Plan’, complete with information
packet and video on the life of Christ’.12 As David S. Olson notes in his recent account of the American Family
Organisation’s campaign against The Last Temptation of Christ, the fundamentalist
protest was effective. Three Republican Congressmen attempted to pass a
resolution that the film be withdrawn from circulation, and the film lost money
at the box office because many cinemas refused to screen it. (It recovered a little
over half of its production costs).13 Indeed, only cinemas associated with MCA
would actually screen the film.14 It is still illegal to watch the film in Chile, and
when the film was released on video, the Blockbuster chain refused to carry it. Controversies surrounding the film were re-ignited in the mid 1990s when it
first appeared on national television in Britain and Canada. In fact, according to
the Independent Television Commission, The Last Temptation held the record
for the most complaints received by a television station in 2003. 7
The body of Christ: Blasphemy as a
necessary transgression?
Carolyn D’Cruz and Glenn D’Cruz This record of
1554 complaints was broken only recently in June 2005, when Jerry Springer —
the Opera attracted more than 15,000 complaints when aired on the British
Broadcasting Association.15 Clearly, when the Christian Right embarks on a
campaign to censor particular representations of Christ, one thing is certain:
millions of Christians throughout the world will dismiss texts that they have
never seen, even though they might be spiritually and theologically engaging. So, why did the fundamentalists protest so much? What incited one Reverend
Bright to describe the film as ‘absolutely the most blasphemous, degenerate,
immoral, depraved script and film that I believe it is possible to conceive’?16 To begin with, it seems that representing Christ with the fascinating possibility
that he too struggled with doubts, fears and passions, like other human beings,
was far more shocking than the supposed ‘true’ flawless Christ told through the
Bible’s gospels. The irony is that this overtly fictional representation of Christ
allows audiences to grapple with Christ’s story in ways that traditional Hollywood
depictions of Christ do not. As Jonathon Rosenbaum puts it: ‘serious engagement
with doctrine is considered blasphemy, yet Hollywood representations of Biblical
incidents are viewed unproblematically, even though they are dependent on a
series of almost arbitrary and often absurd representational conventions’.17 97 Negotiating the Sacred II Predictably, the fundamentalists also objected to the presence of women at
the Last Supper. But more than anything else, they objected to the portrayal of
Christ as having sexual impulses. He is depicted, in the infamous ‘dream
sequence’, as making love to Mary, sister of Lazarus. Notwithstanding the fact
that this sequence is presented as Christ’s last temptation — the desire to produce
a family and make a life with them, as so many other human beings do — the
Christian Right was intent on preserving Christ’s call to sacrifice himself for the
sins of the world as completely unequivocal. If the presentation of a doubting
Christ makes this film blasphemous, then we certainly do believe that blasphemy
is a necessary transgression for redeeming some of the sins of the world today. For without having doubt about the sanctity of our calls — without pausing to
ask how we might distinguish the calls of God from those of the Devil — what
sort of responsibility would we be taking for our ethical behaviour? 7
The body of Christ: Blasphemy as a
necessary transgression?
Carolyn D’Cruz and Glenn D’Cruz Yet, in doing so, we are
forced to reckon with all the atrocities that occur in the world today in the name
of justice. Now, while we might not have the means or power to expropriate the
term back from its current world stage appropriators, we do have the ability to
respond to a certain spirit of justice that ties Christ’s message to what Derrida,
after Levinas, situates as the ethical call of the Other. For Levinas, ethics inheres
in the face-to-face relation, where the self stands accused by the face of the other
before its own concerns with self over-coming. It is the ‘nakedness of the face’
that confronts the self as a summons to responsibility in the encounter of the
other’s death. Levinas wrote: The other man’s death calls me into question, as if, by my possible future
indifference, I had become the accomplice of the death to which the
other, who cannot see it, is exposed; and as if, even before vowing myself
to him, I had to answer for this death of the other, and to accompany
the other in his mortal solitude. The other becomes my neighbour
precisely through the way the face summons me, calls for me, begs for
me, and in doing so recalls my responsibility, and calls me into
question.19 This ethical relationship to the Other is not subsumable within a universal ethics,
as it is intrinsically a singular relation. Unlike Kant’s categorical imperative or
Mill’s utilitarianism, Levinasian ethics do not have recourse to rules and maxims
that guide behaviour. Like the Pharisees who Jesus castigates in the gospels
(Matthew 23:23, Luke 11:42), contemporary fundamentalists promote strict
adherence to the literal ‘truth’ of the Bible, thereby invalidating interpretations
of the holy scriptures that emphasise the practice of justice over sanctimonious
observance of the law. Without detailing the intricate dialogues between Derrida and Levinas
regarding how the Other gets situated in the language of ontology,20 let us focus
on the fact that Derrida reinforces this singularity of the ethical relation when
he distinguishes justice from law.21 For while law aims to be universal and
rule-bound, answering the call for justice must attend to the singularity in which
each of its calls are made. Moreover, Derrida situates law as something that is
deconstructible, because it is institutional, conventional, calculable, legible and
to use a famous Derridean term, ‘iterable’. 7
The body of Christ: Blasphemy as a
necessary transgression?
Carolyn D’Cruz and Glenn D’Cruz This is not to suggest a nihilism or relativism in the making of ethical
decisions. It is rather to acknowledge that ethical decisions are caught within
conceptual distinctions such as Go(o)d and (D)evil, which, like all antithetical
values, are not independent and complete entities in themselves, but knotted
together in the very acquisition of their meaning.18 While Nietzsche exploits
the exposure of the conceptual complicity between Good and Evil in his campaign
against morality, thinkers like Derrida and Levinas, as we will see below,
explicitly link the non-totality of conceptual opposites to the opening toward
the Other. This opening toward the other is known as the ethical relation. The
Other for Derrida and Levinas cannot be known in advance, so in order to be
responsive to the call of the Other we must endure the passage of suspending all
reliance on a programmatic, universalised morality. The scandal over the film clearly illustrates that responding to the Christian
heritage is governed not so much by who hears the call, nor even who answers,
but whose voices acquire power to not only name the call, but act on its behalf. This kind of fundamentalist response to art as outlined above begs a terrifying
comparison with George Bush’s recent proclamation that ‘God told me to invade
Iraq’. For what Bush shares in common with the fundamentalist Right who
frequently sabotage particular representations of Christ is both a forcing and
forging of who gains propriety over the Christian heritage. Crucial differences
between artists as opposed to the fundamentalists and Bush are situated in the
fact that the former explicitly portray an interpretation of their own singular
relationship to Christ without claiming their representations as Gospel, as it
were. Both the Bush administration and the Christian Right act as if they are the
‘chosen people’ responding to the call of God. We are asking here today, if all
of us here and now, whether Christian or not, or whether from any other religious
denomination for that matter, are obliged to take responsibility for what takes
place in the name of such a call. 98 The body of Christ: Blasphemy as a necessary transgression? As mentioned earlier, if there is a call from the Christian heritage that we
want to reaffirm here and now, it is a call for justice. 7
The body of Christ: Blasphemy as a
necessary transgression?
Carolyn D’Cruz and Glenn D’Cruz Justice on the other hand, is ‘infinite,
incalculable, rebellious to rule’ and undeconstructible. For Derrida, maintaining
the space between law and justice is necessary in order for the singularity of
the Other to be given room to interrupt the universality and generality of
calculated laws. In other words, the decision between what is just and unjust is
never insured by a rule as justice comes in the form of a singular idiom. As John
Caputo,22 has noted, Jesus is exemplary here. Through the gospels that tell us 99 Negotiating the Sacred II Negotiating the Sacred II of Jesus making the blind man see, of saving the whore from being stoned to
death, and of raising Lazarus from the dead, we find the ethical resonance of
the face-to-face relation, where Jesus attends to the Other’s command each time
he is called: singularly. In presenting Jesus in all his human frailty (yet without discounting his
divinity), Scorsese opens a space from which we can perceive ourselves struggling
over the receiving end of a call. Who’s calling you? The Bush alliance? The
casualties of war? Or your neighbour in her everyday struggles? Do you respond
to a programmatic, universal ‘Ethics’, or are you finding yourself increasingly
attuned to countless calls from singular others who suffer? These questions are
not supposed to be easily answered. As outlined in what Derrida calls the aporias
of justice, our role in judging the answers to these questions cannot take place
by a mere instrument that calculates. Each decision in the name of justice requires
a ‘fresh judgment’ to be made in a ‘reinstituting act of interpretation’.23 If we
are serious about responding to the heritage of Christ, then this requires a unique
interpretation that no ‘coded rule can or ought to guarantee absolutely’.24 In presenting Jesus in all his human frailty (yet without discounting his
divinity), Scorsese opens a space from which we can perceive ourselves struggling
over the receiving end of a call. Who’s calling you? The Bush alliance? The
casualties of war? Or your neighbour in her everyday struggles? Do you respond
to a programmatic, universal ‘Ethics’, or are you finding yourself increasingly
attuned to countless calls from singular others who suffer? These questions are
not supposed to be easily answered. 7
The body of Christ: Blasphemy as a
necessary transgression?
Carolyn D’Cruz and Glenn D’Cruz As outlined in what Derrida calls the aporias
of justice, our role in judging the answers to these questions cannot take place
by a mere instrument that calculates. Each decision in the name of justice requires
a ‘fresh judgment’ to be made in a ‘reinstituting act of interpretation’.23 If we
are serious about responding to the heritage of Christ, then this requires a unique
interpretation that no ‘coded rule can or ought to guarantee absolutely’.24
Furthermore, a free decision must go through the ordeal of the undecidable,
which is not merely the tension between two decisions, but the negotiation with
that which is beyond calculation; an impossible decision. If Christ did not struggle
between his divine calling and life on earth, there would have been no sacrifice
at all, but the mere robotic motions toward a destiny that had no business with
free will. Yet the passage through the undecidable does not mean that we
therefore forgo the making of the decision; this would reduce action to an equally
inert disposition. The final aporia of justice that Derrida outlines concerns the
‘urgency that obstructs the horizon of knowledge’.25 The immediacy and
urgency in which incoming calls from others are received demand that a decision
must be made no matter how much time and information one has at one’s disposal. We must respond and act in the here and now as Christ responded to the
downtrodden then and there. Such a treatment of justice — the call for a fresh
judgment, the ordeal through the undecidable and the urgency of the decision
— is a far cry from the programmatic doctrine of Christian fundamentalism. If
the struggles of Scorcese’s Jesus give us a character that is more relatable and
accessible to our own ethical problems today, then perhaps the blasphemy
Scorsese is accused of is a necessary transgression for reacquainting ourselves
with Jesus as an ethical activist. As an accessible character, Scorsese’s Jesus also
invites us to ask where our own ethical calls come from, and judge what
temptations we might succumb to, and what sacrifices we might make as we
attempt to heed the calls. For, without struggling with these questions and
agonising over our responses to them — as Scorcese’s Jesus did — we would
be fleeing from a responsibility for passing on the heritage that is committed to
bringing the good news, so to speak, to the oppressed. The spirit throughout the world, the whole of the world The spirit throughout the world, the whole of the world Endnotes 1 ‘Go tell every one’ was sung at folk masses in the UK in the early seventies. We are unable to trace
the author of the song. 1 ‘Go tell every one’ was sung at folk masses in the UK in the early seventies. We are unable to trace
the author of the song. 2 Martin Scorsese in David Thompson and Ian Christie (eds), 1990, Scorsese on Scorsese, London: Faber
and Faber, p.120. 2 Martin Scorsese in David Thompson and Ian Christie (eds), 1990, Scorsese on Scorsese, London: Faber
and Faber, p.120. 3 Rolando Caputo, 1989, ‘Forbidden Christ’, Cinema Papers, vol. 71, p. 7. 4 4 David Thompson and Ian Christie, op. cit., p. 117. 5 Debates about authorial intentionality have a long history and emerge in various forms in discourses
of philosophy, aesthetics, literary theory, psychoanalysis, Marxism and poststructuralism. Our own
relation to the concept of intentionality is guided by Foucault’s ‘What is an Author?’ and Derrida’s
‘Signature Event Context’. Neither philosopher denies intentionality its place in contributing to a text’s
meaning, but both question the ability of intentionality to act as final arbiter of interpretation. Michel
Foucault, 1977, ‘What is an Author’, in D.F. Bouchard (ed.), Language, Counter Memory, Practice: Selected
Essays and Interviews by Michel Foucault, D. F. Bouchard and S. Simon (trans.), Ithaca, New York: Cornell
University Press, pp. 113-38. Jacques Derrida, 1988, ‘Signature, Event, Context’, in G. Graff (ed.),
Limited Inc, S. Weber and J. Mehlman (trans.), Evanston, Illinois: Northwestern University Press, pp. 1-23. University Press, pp. 113-38. Jacques Derrida, 1988, ‘Signature, Event, Context’, in G. Graff (ed.),
Limited Inc, S. Weber and J. Mehlman (trans.), Evanston, Illinois: Northwestern University Press, pp. 1-23. 6 Martin Scorsese in Thompson and Christie, op. cit., p.124. 7 Jonathan Rosenbaum, 1988, ‘Raging Messiah: The Last Temptation of Christ’, Sight and Sound, vol. 57, no. 4, pp. 281-82. 7 Jonathan Rosenbaum, 1988, ‘Raging Messiah: The Last Temptation of Christ’, Sight and Sound, vol. 57, no. 4, pp. 281-82. 8 American Heritage Dictionary. 9 8 American Heritage Dictionary. 9 9 Donald Wildmon with Randall Nulton, 1989, The Man the Networks Love to Hate, Wilmore: Bristol
Books, p. 194. 9 Donald Wildmon with Randall Nulton, 1989, The Man the Networks Love to Hate, Wilmore: Bristol
Books, p. 194. 10 L. Keysen, 1992, Martin Scorsese, New York: Twayn, pp. 166-186; this quotation, p. 168. 11 Wild
it
200 10 L. 7
The body of Christ: Blasphemy as a
necessary transgression?
Carolyn D’Cruz and Glenn D’Cruz So, just as my father sent me 100 The body of Christ: Blasphemy as a necessary transgression? Now I’m sending you out to be Now I’m sending you out to be Section III Multiculturalism and religious pluralism pose a particular set of political issues
for a liberal society. On one interpretation, all that freedom of religion requires
is freedom from persecution. Yet, a stronger requirement suggested for many is
that a liberal society should make allowances for the religious group. For instance,
it has been argued that allowances may be made to enable the member of a
religious group to participate in religious observance, and to maintain their
integrity in meeting religious duties. An even stronger claim is that tolerance
in a society with religious pluralism requires more than mere exemptions to
laws, but respect for their beliefs. As Peter Jones (1990) puts it, ‘“respect for
beliefs” is a principle that is especially relevant to a pluralistic society in which
different groups of people hold fundamentally different beliefs. It holds that,
in such a society, not only should people be allowed to conduct their lives in
accordance with their most deeply held beliefs, they should also not have to
endure attacks upon those beliefs’. Such a principle, however, is seen as deeply
at odds with the liberal principle of freedom of expression. Providing an analysis of what he calls the ‘monologue of Liberalism’, Jasdev
Singh Rai presents the perspective of the Sikh community that was involved in
protests over the play Behzti. Rai rejects the dichotomy between freedom of
expression and religious fundamentalism and orthodoxy implicit in Nash’s
interpretation of the dangers of multiculturalism. What Sikhs were protesting
about was not blasphemy, but the misrepresentation of their religion from the
perspective of the West, and the deliberate intention to offend. Rai makes a case
for the Indic model of tolerance. Plurality of meaning does not exist in Indic
civilisation as two separate entities but subsumed as a whole—what might seem
like conflicting dichotomies to the Western observer, like the sacred and the
profane, coexist without any problem. Conflict arises when attempts are made
through art to impose a hegemonic view on individuals or when what is cherished
as sacred by a community is publicly discredited or desecrated. To bring about
change in what some may perceive as oppressive dogmatism, one does not need
to blaspheme or offend a religion, but rather resort to intellectual debate and
critique. Jeremy Shearmur explores the limits of J. S. Mill’s justification of freedom
of speech in terms of the interrogation of religions’ claims to truth. Endnotes Keysen, 1992, Martin Scorsese, New York: Twayn, pp. 166-186; this quotation, p. 168. 11 11 Wildmon, op. cit., p. 200. 12 Majorie Heins, 1993, Sex, Sin and Blasphemy: A Guide to America’s Censorship Wars, New York: New
Press, p. 167. 13 David S. Olson, 2001, ‘The New Religious Right Versus Media Wrongs: AFA Fights Temptation’,
Journal of Film and Video, vol. 53, no. 2/3, p. 6. 14 Steven C. Dubin, 1992, Arresting Images: Impolitic Art and Uncivil Actions, New York: Routledge,
p. 92. 15 ‘Springer Protests pour in to BBC’, BBC News (Online), 6 January 2005. h
//
bb
k/ /h /
/
d
d /
( Springer Protests pour in to BBC , BBC News (Online), 6 January 2005. http://news.bbc.co.uk/1/hi/entertainment/tv_and_radio/4152433.stm. (Viewed 31 October 2005.)
16 http://news.bbc.co.uk/1/hi/entertainment/tv_and_radio/4152433.stm. (Viewed 31 October 2005.)
16 16 Wildmon, op. cit., p. 201. 17 Rosenbaum, op. cit., pp. 281-82. 18 Friedrich Wilhelm Nietzsche, 2003 (1886), Beyond Good and Evil: Prelude to a philosophy of the future,
London: Penguin. 19 Levinas, Emannuel, 1989, ‘Ethics as First Philosophy’, in S. Hand (ed.), The Levinas Reader, S. Hand
(trans.), Oxford and Cambridge: Blackwell., pp. 75-87; this quotation, p. 83. 20 20 For an introduction to the dialogue, spanning near to thirty years, between Derrida and Levinas,
see S. Critchley, 1992, The Ethics of Deconstruction: Derrida and Levinas, Oxford and Cambridge:
Blackwell. 21 Derrida, J. 1992, ‘Force of Law: The Mystical Foundation of Authority’, in D. Cornell, M. Rosenfeld,
D. G. Carlso (eds), Deconstruction and the Possibility of Justice, M. Quaintance (trans.), New York and
London: Routledge, pp. 3-67. 22 John Caputo, 1993, Demythologizing Heidegger, Bloomington, Indianapolis: Indiana University Press,
p. 205. 23 Derrida, 2002, ‘Force of Law’, op. cit., p. 23 24 Ibid., p. 23. p
25 Ibid., p. 24. 25 Ibid., p. 24. 101 Section III While
supporting Mill’s position, Shearmur offers arguments against an expansive
interpretation of the requirements of freedom of speech. He takes the position
that offence is a harm to be avoided. More specifically, Shearmur advances a 103 Negotiating the Sacred II (tentative) theory about how we might classify ideas, in terms of the degree and
kind of criticism to which they might be subjected, and also suggests that those
of us who operate in a sphere in which ideas are routinely submitted to a high
degree of critical scrutiny, should legitimately exercise a degree of paternalism
— in the sense of not necessarily treating other people’s ideas in the manner in
which they are, prima facie, advancing them, unless they are fully aware of what
the implications of so doing may mean. Typically, people have beliefs that they
implicitly take to be correct and organise their lives by. In going about their
lives, they are not offering their ideas up for public scrutiny in terms of truth
and falsity. He argues that we are only entitled to challenge them when we are
invited to do so, or when such views impinge significantly upon us or those
about whom we care. This suggests that the acceptability of vigorous attacks
upon, or questioning of, people’s views is context dependent. The implication
of the argument from offence is that cartoonists and artists do wrong when they
ridicule religious beliefs, or use them as the basis for ‘aesthetic play’. (tentative) theory about how we might classify ideas, in terms of the degree and
kind of criticism to which they might be subjected, and also suggests that those
of us who operate in a sphere in which ideas are routinely submitted to a high
degree of critical scrutiny, should legitimately exercise a degree of paternalism — in the sense of not necessarily treating other people’s ideas in the manner in
which they are, prima facie, advancing them, unless they are fully aware of what
the implications of so doing may mean. Typically, people have beliefs that they
implicitly take to be correct and organise their lives by. In going about their
lives, they are not offering their ideas up for public scrutiny in terms of truth
and falsity. Section III He argues that we are only entitled to challenge them when we are
invited to do so, or when such views impinge significantly upon us or those
about whom we care. This suggests that the acceptability of vigorous attacks
upon, or questioning of, people’s views is context dependent. The implication
of the argument from offence is that cartoonists and artists do wrong when they
ridicule religious beliefs, or use them as the basis for ‘aesthetic play’. 104 8
The monologue of liberalism and its
imagination of the sacred in
minority cultures The depth and breadth of freedoms in Indic civilisation have evolved over a
much longer period than the experience of freedoms in the West and need to
be appreciated in the context of different political and social philosophies that
have been prevalent in South Asia. A principle perspective in the West is a dualist worldview, and inherent in
Western civilisation are the concepts of evangelic universalism in almost all
fields. A driving force within the imposition of Western philosophical paradigms
in the public space is an assumption that at any given time some basic principles
or ideas occupying the public space in that particular period are universal without
contest: that they are absolutist, from which people deviate at their peril, or the
standard toward which others need to strive. Usually only one philosophical
paradigm or conceptual framework dominates and determines the public space. For instance, in medieval times, Christian dogma was the universal absolute
truth, while post-Enlightenment, secularism and science are the paradigms of
truth. It is in this secular paradigm that freedom of expression is often held as
an absolute, and from which the purist accuses people of deviating from, or of
compromising.1 It is within this context that the sacred and the profane exist
in Western civilisation and the contest between the artistic and the restrictive
is played out in various forms of art. In this binary world, the Church once
considered it a triumph to uphold laws against blasphemy, hence restricting
freedom of expression. The opposite has now been legitimised in the public
space, where the theatre’s efforts to push the boundaries of expression even
through offence2 are seen as a triumph against the traditional restrictions of the
Church. However, to impose this Christian, Western dynamic upon other civilisations
and cultures is at best naive, and perhaps constitutes a political statement. There
is a fundamental difference between Western and Indic civilisations. The essential
philosophy of Indic civilisation destroys generic dualism and absorbs it. It enables
multiple philosophical paradigms and conceptual frameworks to occupy the
public space at the same time. It also abhors evangelic and assertive universalism
and perpetually deconstructs it. Moreover, most Eastern philosophies devalue
anthropocentrism. Since dualism is destroyed, the dichotomy of sacred and
profane has little relevance in Indic cultures. Therefore, the appreciation of the
‘sacred’ has to proceed within different perspectives. 8
The monologue of liberalism and its
imagination of the sacred in
minority cultures Jasdev Singh Rai The particular evolution of Western civilisation has forced upon its artists a
responsibility that becomes invalid and obscurantist when transposed upon the
cultures of Indic civilisation. The fundamental distinctions between the two
civilisations offer different challenges to the artist. But Western hegemony in
the arts often clouds critical thinking and encourages the generalisation of
inherited western experiences and concepts as universal and transcendent rather
than as particular to its own cultural evolution. The arts and the theatre have
enjoyed an uninterrupted progress in Indic civilisations for over 3000 years. In
that period, subjects such as freedom of expression, boundaries, and sacrilege
have been considered in this ancient and pluralistic civilisation, and even
articulated in the ancient text of arts and theatre, Natyashastar. What does the sacred mean in other civilisations, and can it be treated in the
same context as the sacred in Western civilisation? In other words, firstly, can
civilisations be treated to similar artistic explorations, and can art have
intrinsically a uniform role across cultures and civilisations? And secondly, can
the Western format of the institution of arts and its temple, the theatre, enjoy
the same immunity from the normal etiquettes of engagement in Indic cultures
as it does in Western civilisation? The editors’ comment: ‘In recent years, issues surrounding the rights of
minority cultures to recognition and respect have raised new questions about
the contemporariness of the construct of blasphemy and sacrilege. Controversies
over the aesthetic representation of the sacred, the exhibition of the sacred as
art, and the public display of sacrilegious or blasphemous works have given rise
to heated debates and have invited us to reflect on binaries like “artistic and
religious sensibilities”, “tolerance and philistinism”, “the sacred and the profane”,
“deification and vilification”.’ This offers an opportunity to open Western
discourse on freedom of expression to a different construct of critique in which
words such as ‘blasphemy’ lose their meaning and concepts such as binaries are
uncomfortable juxtapositions. There is a paradox in the parody of the passionate 105 Negotiating the Sacred II Western artist struggling against the fabricated demons restricting freedom of
expression that he/she assumes exist within ancient civilisations such as Indic. 8
The monologue of liberalism and its
imagination of the sacred in
minority cultures This chapter argues that
the ‘sacred’ is the creative in Indic civilisations, standing independent of the
concept of the profane; and offence is seen as the failure of theatre rather than
its triumph over restrictions. Moreover, protection of what is ‘sacred’ from
offence is part of the embedded political philosophy of pluralism. 106 The monologue of liberalism and its imagination of the sacred in minority cultures The British play, Behzti, which attracted considerable indignation from the
Sikhs in Britain and was ultimately forced to close down, was set within the
contesting binaries of western philosophy. Its producers had assumed that the
Western conceptual and historical evolution of freedom of expression as a
universal norm is absolute. They failed to see that the play had transgressed the
boundaries of success and entered the arena of failure, as well as the hegemonic
politics in their perception of Indic societies as operating within the same binaries. Principles and pluralism in Indic civilisation As a generalisation, the bulk of Eastern philosophy, with some exception such
as the Nyaya and Vaisesika schools,3 absorbs and devours dualism within most
of its strands. The perpetual conflict for hegemony between opposing
universalisms becomes redundant in Indic civilisation. The artist is free in Indian
civilisation, because the civilisation itself is essentially free from dogma. This is
the obvious thing that escapes the Western liberal artist when Indian cultures
are ‘orientalised’ and treated under the rubric of the dualist struggle in terms
of which the main body of Western civilisation comprehends reality. Edward
Said, who introduced the word ‘Orientalism’4 in relation to perceptions about
the Islamic world, charged the West with failing to understand other cultures
and of imposing its own worldview to make sense of others; this
misunderstanding was partly romantic and partly demeaning. Such an approach
perverts the understanding of alternative cultures, and eventually it leads to
the leading Western-educated minds in other cultures internalising the myth
created by the West of themselves. Similarly, Western liberalism has generally
failed to comprehend the non-dualist world view of the East in practical everyday
life. Although many Western intellectuals romanticise it ‘intellectually’, they
rarely understand its practical expression, and often simplify the seeming
contradictions in Eastern cultures in Western dualist contests of good–bad,
freedom–oppression. Multiple and sometimes competing complexities fusing into one strand are
a feature of the dominant Vedanta school of which the Advaita (meaning non
dualist) Vedanta5 of Adi Sankara6 has inspired most of Hindu thinking. In
Vedic thinking, there is not only Brahma the creator, and Shiva the destroyer,
but also Vishnu the preserver. One creates, the other destroys, the third
preserves. What is destroyed is created again, preserved and destroyed and it
goes on. Time and space become irrelevant, as does history. There is harmony
among the three rather than conflict. Underlying all this is the still, unmoving
Brahaman,7 the Eternal in whom everything finally collapses into, who has set
this drama, the Lila, an illusion of cycles perceived subjectively by each human
mind. Thus, Hindu philosophy obfuscates certainty and undermines
fundamentalism anchored in divine revelation. 107 Negotiating the Sacred II In the ordinary tapestry of Indian life, Rama, the god of virtue and charity,
can sit with Kali, the god of destruction and sacrifice. Principles and pluralism in Indic civilisation The Dharmasashtra, the
text of chants of moral duties can be read with the Kama Sutras, the texts of
pleasure, in the same temple. Varanasi, the holiest place of worship on the Ganges
is not very far from Khajurao, where the temples have bestiality, orgies,
homosexuality and heterosexual positions carved on temple walls. The sacred
and profane are not seen as in conflict with each other. The tapestry is flattened,
all existing in the same domain, the same corner, walking and breathing together
without threat from the other or threatening the other. In the Sikh philosophy, Satguru, the eternal teacher is called by so many
attributes and character names that the name loses meaning, becoming ‘anaame’8
(without name) in the Dasaam Granth, the text written during the period of the
tenth Sikh Guru, Guru Gobind Singh. The anaame is the benevolent architect
of the tranquillity of a quiet beach, and also the alleged demonic destroyer who
created the Tsunami9 as explained in the Guru Granth Sahib, the textual guide
of the Sikhs compiled by the ten Sikh Gurus.10 Everything functions within
hukam,11 the laws, but no one quite understands why things happen,
ultimately.12 Bad and good are in people’s minds. We can venture conjectures
on the mechanics, but to understand ‘the why’ is beyond ordinary language
and discourse. No one can know the ultimate truth; therefore no one can claim
to have knowledge superior to that of others.13 Dialogue and critical discourse is an intrinsic feature of Indic philosophies. Consequently Indian philosophies and cultures are used to disagreements and
critical views. But in critique and dialogue, offence is avoided in Indic cultures. For instance, within Sikhi, the trinity of Hinduism (called Trimurti) is not
dismissed with a language of condemnation, but it is made irrelevant through
subtle critique.14 The human can outwit the trinity gods and make a direct
connection with the Eternal. The pantheon of Hindu gods is thus made irrelevant. But, in the Sikh scripture, Sri Guru Granth Sahib, the language is never offensive,
never commanding, just as it never is in Indic traditions. It initiates a debate
within the individual to come up with the desired inference. The one eternal
divine reality of Sikhi and the multiple gods of some sections of Hinduism have
coexisted in the same territorial, social and political space without either system
waging a crusade against the other. Principles and pluralism in Indic civilisation The sacred and profane exist within the same context, in the same dimension,
in the same space. They are not demarcated by territory or barriers. Differences
exist in people’s minds. Therefore, there is no battle for the artist to overcome
or external forces to challenge, except people’s minds. The success of the artist
is to introduce a dialogue within the mind. But this is where the Natyashastar 15 places sanctions, as does Kautaliya’s
Arthashastra. Both prohibit deliberate offence. The reason becomes apparent 108 The monologue of liberalism and its imagination of the sacred in minority cultures when other aspects of Indian civilisation and the theory of arts are considered. The Natyashastar, written about 2500 years ago, is the text of theatre and arts. The Arthashaster, also written around the same time by Kautaliya, is a text of
statecraft and politics, written at least 1500 years before Machiavelli’s The Prince. Most Indic philosophies believe in an eternal drama. Creation is engaged in
a drama, the lila (Sanskrit), constructing, deconstructing in perpetual cycles
operating concurrently.16 Lila is the key concept that enables Indian thinker
to escape from universalism and anthropocentrism. There is nothing special
about Man. All creation is engaged in this perpetual drama. Some day, humans
will become extinct and may rise again. All human ideas are conjectures as no
one can penetrate the ultimate reality of the drama. On the other hand, our
perceptions are illusions, maya, which are real for us, but in the wider context,
part of illusion within the lila. This enables an enduring plurality and the freedom from hegemonic
tendencies. The philosopher is under no compulsion to be an evangelist and
spread his inspirations or impose his world view. In Indian tradition, the guru
(the guide) waits, and the disciple comes to him. Different gurus have different
perspectives. The Nyaya school17 is highly analytic, as opposed to the intuitive
and even blind belief system of some schools. It uses principles of human reason
to deconstruct. The Samkhya school is atheist.18 It believes that only what we
can see, feel and reason, exists. The Vedanta, the most popular school in Hindu
society, believes in the Brahaman,19 the creator and the creation as maya, an
illusion. Yet, the different schools co-exist in the same town, and in the same
Kingdom. Stated within the acceptable norms of dialogue, nothing is shocking and
nothing is really suppressed in Indic civilisation. Principles and pluralism in Indic civilisation There is nothing that invites
the wrath of the ‘Church’, as there is no Church. Since there is no concept of the
sacred and the profane as distinct entities, everything is sacred. All life, all
creativity and all action are sacred. Even rejection of a divine order or a God
isn’t considered blasphemous, unholy or a sacrilege. In fact, the Samkhya school
of Hinduism is atheistic and has coexisted as an alternative philosophy of reality
without tension between it and other systems. Hindu civilisation makes a subtle difference between philosophy and
pragmatic everyday life. The differences are manmade. People pray to stone
gods, but the philosopher engages in dialectic beyond gods. Beliefs exist in
perpetual dialogue with others, with the environment, and with the abstract
perception of truth. The individual is free to choose his god and his version of
reality, his method of perception and form of sacredness. The individual’s
engagement is voluntarily. No church or doctrine forces a particular world-view
upon the individual. But the culture imposes that he or she accepts and respects
the personal space of the other. What one individual considers sacred may seem 109 Negotiating the Sacred II ridiculous to the other. This degree of pluralism survives by each person and
community respecting the other for what they are, not what they can be in one’s
terms. In this free-for-all world, order is maintained in society through some
regulation. Civil regulation is part of society, made sacred, but never so
universalist as to warrant crusades to convert other systems. There are at least
four versions of the Dharmasutras, 20 (also named Dharmashastra) the texts of
duties, but there have never been conflicts or wars over ‘which’ Dharmasutra. Whenever domination occurs, it is asserted out of pure desire for power and not
in some vague belief in one single absolutist idea of the cosmos and life. There
are laws, but they are often made by the hegemonic caste or group. These are
not absolute laws. They are enacted and imposed consciously by a section of
society for its own benefit, sometimes hideous and unjust laws which function
to create privilege. The upper caste claims authority from the ancient sages for
these laws and repression of others. But if the Brahmins created the caste for
their own power, the sanyasi (ascetics) made the casteless system outside the
civil. Principles and pluralism in Indic civilisation In traditional India, the towns and villages were often regulated by the
Brahmin with rules codified in Manu’s laws.21 However, the dissenter could
legitimately walk out on this and into the almost bohemian and non-materialist
life of the sanyasi, with their camps and dwellings in forests. The sanyasi could
wander into the towns and villages without fear, but did not stay there to avoid
living under civil laws. One version of the Hindu text, Ramayana was written
by Valmiki, a sudre (of the low caste), yet Brahmins, not so long ago, cleansed
themselves even when the shadow of a sudre prevailed on them. But one almost universalist concept in Indic civilisation is to limit the scope
of gratuitous offence as a weapon of critique, in order to maintain this fiercely
pluralistic society. This is the principle that requires the arts and the theatre to
avoid offence for the sake of offence, ridicule for the sake of ridicule, or
‘profanity’ simply to make a point. The sacred is not universal but individual
to the person or the community. Each person and each community of believers
creates his/her own space outside others. That space is its ‘essential’ and, therefore
sacred. And each creates its own rituals, practices and causes. The rules of
dialectic allow critique but not offence. Offensive acts towards or offensive
representations of another’s sacred is a political statement. It is the equivalent
of robbery, impropriety or intended humiliation. If people were to ridicule the other’s choice, make mockery, deliberately
offend, and purposely destroy the ‘sacred’ imagination of another, society would
break down into a series of violent retaliations and power struggles. This is the
basis of self regulation in Indian civilisation. The distinction between critique
and offence is embedded in culture. Since another’s god is not imposing any
laws upon one’s own value system, there is no reason to embark on a campaign 110 The monologue of liberalism and its imagination of the sacred in minority cultures of humiliating the other’s god or the other’s beliefs. There is no mileage in parody
of the sacred, since the submission to a god or a system is voluntary. This principle of civility allows the atheist of the school of Samkhya to live
as neighbour in the same street as the believer in the elephant god, Ganesh. Principles and pluralism in Indic civilisation The
person who sacrifices to Kali (goddess of destruction) and the Sikh who rejects
this multitude of gods and idol worship and the Muslim who believes in a
revealed religion, the Christian who believes in a son of God and the Buddhist
who does not ponder about a creator, can coexist. This coexistence would break
down if the humanist put up plays offending the others in his or her assumption
of superior values, or the Christian stamped on the elephant god as devil worship,
the Sikh spat on the idol of Kali, and the Muslim tore down the crucifix of Christ
under pretext of freedom of expression. Indian society would simply tear itself
apart. Plurality survives on rejection of universality but respect of the other’s
sacred space. One of most spectacular living examples of this plurality of beliefs,
philosophies and ways of lives is seen at the three-yearly festival of Maha Kumbh
Mela, rotating between four cities in India. The Hindu Pantheon, from the nanga
sadhus (naked holymen) to the rigidly caste-ist and puritanical Brahmins, come
together in a gathering of millions. Nothing creative can shock in the Maha
Kumbh Mela. In this grand spectacle of Indian civilisation in performance, there
is no absolute ceiling, no divinely ordained universal restrictive laws, and no
desecration as understood in Western civilisation. It is a world of believe and
let believe. Perhaps the very concept and terminology of religion is misplaced in Indic
cultures. Hinduism has the Dharmasutras, the texts of natural duties, the
Shilpasashtra, the texts of science and architecture, the Arthasashtra, the text
of politics and statecraft, the Kama Shastrar, the text of pleasure, and the
Natyashastra, the text of theatre and arts as well as many other smaller Vedic
texts. But they also coalesce. There is the wide-ranging Manu’s laws,22 which
deal with individual moral life, inter-personal interactions, community relations,
commerce, politics and in fact most aspects of life. They are part of the so-called
sacred cannons of Hinduism. How can these broad fields be classified as religion? They are part of a civilisation that does not differentiate between the religious
and the political, between the secular and the spiritual. Indic philosophies tend
to work on the premise of a holistic approach and interdependency. In Sikhi too, the distinction between religion and politics, worship and action,
science and spirit, simply do not exist. Principles and pluralism in Indic civilisation They are artificial atomistic distinctions
created by modernity and when applied to Sikhi, they confuse interpretation
and meaning of the Sikh teachings. The word that Indic systems use to describe
themselves is ‘dharma’. 111 Negotiating the Sacred II Negotiating the Sacred II The word ‘dharma’ does not have a direct transliteration in English. Sri
Krishna Kant, Vice President of India in 1997, made a clear distinction between
dharma and religion.23 He described dharma this way: First and foremost it [dharma] means living in harmony with nature and
natural laws. It means to live by moral and ethical principles of the
society without surrendering the freedom to question them. The term
‘Yuga Dharma’ signifies that Dharma itself is continually evolving and
not rigid or inflexible. The continuous evolution of Dharma has been
through debate, and the triumph of logic, consensus and harmony. Most
importantly, Dharma is not linked to any religion or set of beliefs.24 The Krishna society describes dharma as ‘the inseparable quality that makes
a thing what it is. A stone’s dharma is to be hard, water’s is to be wet, fire’s to
be hot, sugar’s to be sweet’.25 The Mahabharat, one of the sacred texts of
Hinduism, describes: (‘dhaaranaat dharmam ityahuh dharmo dhaarayate prajah,’)
‘They call it dharma, since it upholds; it is dharma that upholds the people (of
the world)’. That which upholds, supports or sustains this universe, without
which the universe would disintegrate, is dharma. The Guru Granth Sahib, the living textual Guru (Guide) of the Sikhs does
not use the word mazhab, which translates as ‘religion’, but the word ‘dharam’
(a Punjabi word for dharma) as the context of its teachings. There is now a body
of work in India which refutes the use of the word ‘religion’ in reference to Indic
systems, and which considers the word to be appropriate to Abrahamic traditions,
with a restrictive meaning. The word ‘dharma’, in my own writings is described
as the essence of modern science with the spiritual dimension intrinsic. Thus,
it is the elusive truth which science is trying to search and which Indic traditions
have been speculating and searching for a long time. Religions, on the other
hand, tend to end up in proscriptive truths. (Truth, that is, so-called.) Therefore, parallels with Western civilisation become irrelevant and irrational. Principles and pluralism in Indic civilisation Indic systems never strive to create a single way of thinking or a single ‘truth’
or system of understanding reality. The civilisation is most at ease when there
is pluralism. From this perspective, statements such as ‘binaries like artistic and
religious sensibilities, tolerance and philistinism, the sacred and the profane,
deification and vilification’ lose their meaning and context. The challenge from the universalists Indic civilisation has seen an invasion of its systems at least three times, and all
from the Abrahamic universalist creeds. First Islam crusaded against India’s
idols, and then it embarked on an economic and sometimes oppressive program
of converting people to its revealed set of universals. The Sikhs, the Marathas
and Rajputs (Hindus) rose against the Mughals to stamp out their intolerance. Then Christian zealots and missionaries and neo-scientists came with the Empire, 112 The monologue of liberalism and its imagination of the sacred in minority cultures openly mocking and ridiculing the gods of the Hindus. Mahatma Gandhi used
this offensive and oppressive behaviour of the colonist as one of his campaign
logics to drive out the British. The humanists (an atheist version of Abrahamic
creeds) gained enough Indian converts and caused mayhem through secular
democratic theory (Indian Congress Party) and the communist party. There has
been a tension between the ‘reformists’ and traditional India since independence. But gradually, ancient India is pluralising the liberal universalist. Since all human
thought and ideas are conjectures, the very idea of a set of some secular principles
being more sacred and universal than the ideas and practices guiding worship
of the elephant god is an antithesis of Indic philosophy. More than three thousand
years of cultural pluralism has survived by restraining from imposing. In post-colonial India, modernism with its universalist tendencies, has cast
a long shadow in the social, political and arts fields. It has influenced mainly
the educated classes. Traditional philosophy is not taught in schools, and people
are left to find out about it through religious institutions or general literature. It is only in the last two decades that some critical thinking is emerging on the
different conceptual foundations. It is quite common to find that ‘educated’
Indians think in Western dualist concepts and Western ideas of freedom of
expression. The challenge from the universalists No lesser a person than Vice President Krishna Kant commented on
this: ‘The Western education system forced us to think in Western ways.’26
The most likely reason may be discerned from Lord MaCauley’s words in
February 1835 when he wrote his rationale for a new education system for British
India (during the Empire): ‘We must at present do our best to form a class who
may be interpreters between us and the millions whom we govern; a class of
persons, Indian in blood and colour, but English in taste, in opinions, in morals,
and in intellect.’27 There hasn’t been a fundamental reform of the Indian
education system since. It is common to find the average Indian brought up on
traditional education and knowledge, reacting differently than the educated
classes to similar issues. One of them is freedom within the arts. Interestingly,
a modernised universalist and united but intolerant form of Hinduism has also
emerged from the educated classes which David Ludden ascribes to the influence
of modernity.28 It is expressed as political Hinduism of the Hindu Mahasabha,
an umbrella organisation of Hindu movements which started in the early
twentieth century in reaction to British colonialism, and which gave rise to the
Bharatya Janata Party (BJP). Critique and conflicts within Indic discourse In Indic civilisation critique is encouraged, a dialogue of disagreement is
encouraged and a discourse of rejection of the other’s belief through debate
(textual, verbal, and visual) is promoted. In fact, the Upanishads are critiques
written and discoursed over centuries. However, there is a balance between
offence and critique which has been observed in Indian civilisation for centuries. 113 Negotiating the Sacred II When it breaks down, it leads to violence and disruptions. This became evident
in the modernised version of Hinduism of the BJP, with its intolerant universalist
and united formulation of Hinduism. But ancient India rejects them just as it
dislikes the secularist enforcer. There have been breakdowns and conflicts between different groups in Indian
history. They have been conflicts for power. In the Hindu pantheon, even gods
fight. The gods of some people win wars over the gods of others. The gods of
some are plagiarised. But the culture reverts back to its pluralistic stability and
mutual respect. For instance, the Sikh Scripture, the Guru Granth Sahib subtly
critiques the Hindu pantheon. The Hindus treat the Granth Sahib with reverence. The Granth Sahib does not offend but critiques, thus respecting, but not agreeing
with, the Hindu pantheon. The arts and particularly theatre is a field in which critique and disagreement
has been expressed for centuries. However, this has generally followed guidelines
established by the ancient text, the Natyashastar. Indian arts and theatre, the Natyashastar In ancient Indian arts theory, the entirety of existence is itself theatre. The
constructed theatre of man is only another drama within it. It therefore evolves,
dissolves and plays within the larger theatre of life, ideas and philosophies. The Natyashastar (or Natyasastra) of Bharata (the writer/s), the oldest text
of arts and theatre in India, propounds a sophisticated theory of theatre and of
the performing arts. The Natyasastar stands as one of the greatest books from
the ancient literature of Indian civilisation and brings the theories of the other
shastras into drama. It is a detailed text on the theory, practice and rules of
performing arts. Five main principles summarise the Natyashastar. These
principles are Artha, Rasa, Abhinaya, Sahmita and Natyadharma.29 Artha is the sentiment or the substance that the author wants to convey, that
is, the message of the text. In Indian theatre, the text is extremely important,
written within the wider philosophy of maya (illusion of reality) as lila (drama). It is not absolutist. It engages human life and the wider drama of existence. The
message, the substance, strives to impact through Rasa. Rasa is the essence. Rasa cultivates the receptive faculties of receivers
activating a whole range of responses whose aim is a condition of sahmita
(wholeness). Rasa is not a sudden revelation;30 it results from a gradual
revelation. The performer and the audience become engaged in the whole
performance. But bhavas, or states of mind, have to be aroused. The Natyashastar
recognises two distinct bhavas. The sthayibhavas, 31 of which there are nine,
are dormant in human nature and aroused by external stimuli, and the
vyabhicaribhavas, of which there are 33,32 and which are transient and fickle. 114 The monologue of liberalism and its imagination of the sacred in minority cultures Abhinaya is the variety of props and techniques that give visual form on the
stage. This is the function of the actor and the director.33 Sahmita is the concept of wholeness. A particular event or a story may be
performed, but the story is in the context of the holistic view of life, the
intertwining facets of human existence that are essential in Indian philosophy. Indian arts and theatre, the Natyashastar As Yarrow states from the Natyasastra: ‘Art is not diversion, escape, amusement:
it is means to wholeness of vision and to the integrated function of perception
in more than ordinary modes.’34 Sahmita also concerns the intrinsic knitting
together of artha, the rasa, and abhinaya. Natyadharma, the rules of drama, determine how, when and what can be
put on stage in one form or other. The rules require that the play avoid grim
realism, tragic ends, sexually explicit actions, political revolt and offence.35
Although theatre in India has made some dramatic departures over time, for
instance, plays have put tragic and grim ends (for example, the tragic love operas
Heer Ranjha, Sohni Mahiwaal and the Tamil classic Shilappadikaram 36 ),
contained politically subversive messages (in the dramatic political poetry of
folk singers called Dhaadis in Sikh uprisings against Mughals, and in Nil
Darpan,37 an 1860 play that exposed British colonialism’s effect on indigo
workers and was banned by the British subsequently leading to the Censorship
Act of 1876), and have been sexually explicit ( such as the temple dances of
Devadasis, which shocked the British), the principle of avoiding offence was
retained throughout. However, according to Kautilya’s Arthashastra, the guide
to State craft, entertainers were permitted to make fun of the customs, castes or
families and the practices and love affairs of individuals.38 The fine distinction
between ridicule, fun and gratuitous offence was maintained. The early Indian
cinema stuck close to the five principles of Natyashastar, and modern Indian
cinema still avoids offence. The other aspect of theatre and arts in Indic civilisation is that, unlike Western
theatre, the inherited Roman amphitheatre, it generally lacked distinct theatre
enclosures with an audience captured in a walled setting. The temple itself was
theatre. Theatre travelled to the ordinary person, into the villages, in remote
places and in big cities.39 It blended. The performance could last for days, which made the task of the performers
extremely difficult as they had to retain the rasa (essence) from day to day, keep
the audience engaged, and get the message across. The challenge was to deal
with difficult critique without offending, but leaving a deep impression that
asked the audience to think for a long time after the performers had left. The
play avoided immediate offence, but succeeded in leaving a sentiment and a
dialogue within the recipient. Indian arts and theatre, the Natyashastar 115 Negotiating the Sacred II Thus, the traditional Indian audience is sophisticated, having inherited a
pattern of theatre which rejects universality and sustains the general principles
of plural life in Indian cultures. Traditionally, Indian theatre has been conscious
of the freedom of conscience that allows for a pantheon of beliefs and choices. This gives the artist complete freedom in creativity. Theatre understands that
each person and community creates its own sacred space as its identity and its
creative activity. The creation of these variable and innumerable forms of creative
sacred spaces gives rise to forms of art and expressions of the individual,
deliberately transported out of the domain of explanation or reason in many
instances. Disparate systems co-exist within a healthy dialectic of critique and
doubt. But the art is in learning the boundaries of critique, so the arts never
become offensive. This is understood within traditional theatre, which can be
provocative without being offensive. The play succeeds if it can engage the
audience through rasa, with the artha (the sentiment) that invokes thought
rather than anger. It fails if it causes offence and provokes violent response or
anger. Modernity and Indian arts In this ancient and highly sophisticated civilisation, modernity has created
disruptions. Many Indians educated within Western systems are oblivious of
the history and theories of the Natyashastar, and have transformed Indian theatre
to a mirage of Western theatre. Indian life has been ‘invaded’ by western cultural
forms.40 Theatre is bound by the limits of time and it uses Western theatre
theory to be provocative. But the audience of the villages and the traditions are
used to a different sophistication in theatre. They do not like the brutal form of
theatre of the West. They can appreciate the subtle and entertaining form of
Western theatre. But they react adversely when it provokes with offence, because
this breaks one of the most sacred principles of Indian plurality, ‘respect and
be respected’, the dialectic of critique without the politics of desecration and
hegemony. It is an irony that the Western liberal cannot teach the traditional Indian
freedom of thought as it is intrinsic in Indic civilisation, so he/she reverts to
teaching the Indian freedom to offend! But an important question is ‘offend what
and offend for the sake of what?’ If criticism and disagreement is allowed and
since no system of belief assumes a position of infallibility, what exactly is offence
trying to achieve? To offend implies a self elevated sense of superiority as the racist in the street
does. To offend assumes one knows better than the other. But, according to Indic
civilisation all human knowledge is ultimately a myth, a creation of human
imagination. The truth remains elusive. The Indic civilisation does not support
the belief that there is an objective knowledge out there to be decoded
definitively through the empiricism of science. Even science is a human 116 The monologue of liberalism and its imagination of the sacred in minority cultures construction, a certain way of looking at things, a methodology. In the Indic
context, why is the myth of the liberal any superior than that of his predecessor,
the Christian or the Muslim? And how are the myths of the Muslim any superior
to that of the Christian or the Jew? How are the humanist’s ethics any more real
than those of the animist or the so called superstition of Ganesh, the elephant
god? The humanist calls his creed rational liberalism, the Christian fundamentalist
claims his creed to be a divine revelation that cannot be faulted. Modernity and Indian arts The extremes
of the two exist in tension and compete to claim authentic legitimacy. The use
of the weapon of offence in this duel is understandable as neither concedes
ground or acknowledge the other to have legitimacy too. When the dialectic
breaks down or is restricted, the politics of violence and offence takes over. The new religion of Western civilisation is a certain construction of human
reason, a particular worldview. The liberal is as fanatical as his predecessor, the
Christian believer. Just as the Christian wanted to spread the message across the
world, and force it down the throats of the rest of the unwitting world, the
liberal assumes a similar crusade driven by a belief in the universality of his
ideals. When examined in context, the genesis of Western liberal thought from
the Enlightenment appears to have failed to address the essential problematic
feature of medieval Christian dogma. The sinister fanaticism, the certainty, the
universalism that gave rise to the crusades, colonialism, slavery and oppression,
has crept through intact into the new liberal creed. Negri’s ‘Empire’41 is a great
expose of the neo-colonialism of liberalism. All that has changed is that faith in
the certainty of divine revelation has been replaced by faith in the certainty of
human reason. The West stumbled towards freedom from the oppressive
restrictions of the Church. But it shows little if any recognition that the ‘other’
(that is, Indic civilisation) has had freedom for at least three millennia and has
no sanction against criticism, no system of inviolable oppressive ethics, no
concept of the proscriptive, no problem with freedom of expression and no
concept of the sacred as defined in relation to the profane. The distinction
between sacred and profane does not exist in Indic theatre.42 There isn’t
comprehension that Indic civilisation does not have a sense of religion in the
context of Abrahamic religions. It appears that hegemony seems to empower
interpretation and comprehension of the other through one linguistic tool and
worldview. In the Indian context, the profane is the universalist, because it attempts to
undermine the basic Indian concept of pluralism. When the sacred is deliberately
treated with disrespect, rather than critique, it is seen as the beginning of political
universalism, a concept Indian civilisation has successfully contested and defeated
time and again through its known 3000 years of civilisation. Behzti, the contest of civilisations We come to the practical: the real drama of Behzti. There were at least three
plays in the entire saga: the serious play by Gurpreet Bhatti, the semiotic play
by the Birmingham Repertory Theatre (or its director), and the drama out in the
street — a clash between two civilisations. Bhatti’s play was simple and straightforward. Written from an orientalist
perspective of Sikhi and full of cultural clichés common in liberal fraternity and
the Westernised sector of Sikhs, the play tried to deal with the humiliation of
women in a religious context. Lacking any deeper understanding of Sikhi, she
interposed Christianity as a model of religion upon Sikhi and the Sikh Gurdwara
as the institution. The characters involved the president of the Gurdwara and
people around him. The play’s theme is sexual corruption and financial greed. He is killed by one of his victims. Ms Bhatti admitted that this was pure fiction
and without any basis in reality. The Sikh public is weary of these negative stereotype plays which seem to
be the ones that get financial backing and mainstream opportunity for
production. However, the Sikhs are used to parody and to jokes about them. Indians joke about Sikhs as the English joke about the Irish. The Sikhs even join
in these jokes and joke about themselves. Moreover, fictionalisation of life in
theatre is something Indians (and Sikhs) are used to. The Sikh tendency is to
ignore these, as they initially did with Bhatti. It is possible that some
British-educated Sikhs may have been offended by this type of negative portrayal
production and might have written to the theatre or local council. But that is as
far as the protest would have gone. The majority of Sikhs, born in India, would
simply have brushed off the production. Drama is fiction. There have been
controversial and provocative productions by other young Sikh writers in the
past, but the Sikhs have simply ignored them or found them amusing. A
playwright called Harwant Bains produced a play in which a Sikh cuts his hair
on stage and mocks the ‘religion’. He tried his best to get publicity by provoking
the Sikhs. He went on to claim to have had threatening phone calls, possibilities
of demonstrations, etc. The Sikhs simply ignored him. Fiction is fiction. His
production died a quiet death. Modernity and Indian arts This was at the
heart of the response of the non-Westernised Sikhs in UK to the crude theatre
of Behzti. 117 Negotiating the Sacred II Behzti, the contest of civilisations Moreover, the theme of humiliation of women is not new in Indian theatre,
or to the Sikhs. The attempted disrobing of Draupadi the wife of the Pandavs
in the epic Mahabharata, the sacred Hindu text about dharma and virtue in war,
is a classic narrative in Indian culture. Sikhi, like other Indian philosophies,
makes no distinction between the religious and the secular. Inevitably, a narrative
of a Sikh man humiliating women would involve the principles, or lack of them,
of his dharam (moral duties). Dharam and ‘religion’ are two different words with
different meanings, as described earlier. Indian philosophies call themselves
Dharma and not ‘religions’. But colonialism insisted on a particular category, 118 The monologue of liberalism and its imagination of the sacred in minority cultures and hence forced a dichotomy upon Indic cultures. The dichotomy between the
religious and the secular is meaningless in an Indian cultural context. Indian theatre has dealt with the theme of women’s humiliation successfully
for millennia without offending the audience and yet leaving the message of the
writer deeply imprinted in the minds of the audience. The Sikhs expected that
subtlety from Bhatti. Many traditional Indians expect writers of Indian origin
to know of the traditions in Indian theatre. But disjunctions in cultures and
community become apparent in instances such as this which are a theatre of
their own. Ironically, neo-Western Indians, ingrained with universalist liberalism,
think they are ‘educating’ their ‘provincial’ community while the community
assumes that such Indians have a critical understanding of their civilisation. These conflicting assumptions were evident in Behzti. Behzti went a bit far. Not the play, the artha, written by Gurpreet Bhatti,
but another play in the background. This was the abhinaya playing silently,
whose sole aim was to offend. Abhinaya is the variety of props and techniques
that give visual form on the stage. The props used to give form to the play were
not fiction nor did they have any relevance to the play. It was a semiotic theatre
reminiscent of colonialism, triumphalism and power. This was the director’s
play; the director wanted to provoke for the sake of provoking, offend for the
sake of offending. The director had power, the public’s money behind the theatre,
and a mere two centuries of fundamentalist Western liberal rhetoric. Behzti, the contest of civilisations The director
tried to bring an Indic civilisation system into the dual context of Western
historical experience and the recently discovered freedom of expression. The
director tried to construct the concept of sacred and profane, the triumph of
reason over ‘religion’, an assertion of crude freedom over conjectured proscriptive
restrictions. The director tried deflating the icon that is central in the Sikh system
and tried to make the essential in Sikhi mundane and irrelevant. The director
was on his own little crusade to teach the Sikhs about freedom of theatre and to
gain publicity in his attempt. But, for the Sikhs, the play had gone beyond rasa
into offence. It was no longer attempting to stage Bhatti’s play, but the Director’s
perversion. The artha, the rasa and the abhinaya had no cohesion, or sahmiti. As far as the Sikhs were concerned, it was no longer theatre but the director’s
‘safe zone’ of intentional humiliation: the offence that was disallowed in
Natyashastar. This was no longer theatre nor a creative act, but a political
statement. From their perspective, theatre had failed and entered into the politics
of neo-colonialist presumption. The Sikhs, like most Indians, are sophisticated in receiving theatre. Thousands
of years of cultural orientation are not lost merely by migrating to another
country. Controversy, criticism, subtle deconstruction, bad language, sexual
perversions, character assassination, etc, are all acceptable as part of fiction, but 119 Negotiating the Sacred II political humiliation and deliberate offence is not acceptable. It broke the
natyadharma and failed to create rasa. The director brought the Gurdwara and the Guru Granth Sahib’s text into
the stage where rape and corruption were being staged. From an Indian
perspective, this was no longer theatre, but a political statement. At best, it was
a misguided political statement imagining a battle between artist’s freedom and
religious dogma. This has no history or relevance in Indic civilisation. Indian
civilisation recognises the artist’s total freedom, but also the artist’s responsibility
towards wider pluralism and the freedom of the individual to his sacred art. It
was also clear from the play that the writer had no real knowledge of Sikhi, the
institution of Gurdwara, or the benign teachings of Sri Guru Granth Sahib, the
living textual guru of Sikhs. Behzti, the contest of civilisations The director treated the living created iconology
of the Sikhs into his dead world of intellectual discourse, the post enlightenment
war between the Western concept of human reason and its assumed irrelevancy
of the Abrahamic God in Western public life. At best, it was ignorance of the
‘other’ because there is no such conflict in Sikhi or Indic civilisation. Sikhs protested. It is interesting that on the twelfth day, British born
non-practicing young Sikhs emerged from the pubs in Birmingham, saw the
pain of their community and took matters into hand. They attacked the theatre. Although brought up in Britain, they do not share the British middle class
reverence for the Western theatre as a ‘sacred’ place. The theatre stage has
replaced the Church pulpit in the West. At one time in Western history, the
pulpit was the safe stage from which war could be waged against ideas of heresy
and blasphemy. Now it is the theatre which is the safe stage from which war
can be waged against the very concept of ideas being called heresy or blasphemy. But this historic Western conflict between Church and State, between Church
dogma and scientific thinking, between Christian proscription and Western
liberalism, the perpetual tension of duality, receives little empathy within the
civilisation of Indians. Indians have inherited a tapestry of plurality of coexisting
imaginations without restriction, and in the minds of Indians, theatre is the
drama beyond the confines of the building. The ‘magic’ ‘epic’ or ‘divine’ world
co-exists with the everyday (Yarrow).43 In the Indian mind, there is nothing
sacred about theatre if it breaks from the natyadharma (the rules of engagement
in theatre). Once a performer moves away from dharma, he/she forfeits the
assumption of sacred ‘freedom’. The Gurdwara as the ‘sacred’ Since everything is maya, an illusion of realities that emerge and then vanish,
drama is played in the context of the wider world. According to Sri Guru Granth
Sahib: ‘This world is an illusion; it dies and it is re-born — it comes and it goes
in reincarnation.’44 The Sikhs believe that every second is a new creation. The 120 The monologue of liberalism and its imagination of the sacred in minority cultures past dies, its memory only constructed, the present is real for a nanosecond and
the future hasn’t arrived, it is just conjecture. The Hindus carry out a special service called Arti. It is worship of the
creativity of nature but represented in a plate. Arti is very sacred. The Sikh
gurus critiqued this. Arti is the entire world. Everything is in a creative state,
dying and reliving. The individual needs to break from the confines of buildings
into the broader space that exists around him or her. The sacred space is the
wider world, not in a confined theatre or building. But then, why is the Gurdwara sacred? The Gurdwara is not sacred in the
sense of demarcated territory. It is the sovereign territory of Guru Granth Sahib,
a text to other people, but which is treated as a human person when it is read. Derrida’s philosophy of the differ/ance may help to explain this phenomenon. The Guru Granth Sahib was written by the Sikh Gurus themselves and is in
raags (musical format) and cannot be changed, added to or modified. The text
is considered as the spoken word without the lag that Derrida conjectures,
without the censor of the human mind, without that corruption that takes place
between the hidden and the stated. One of the practices in Gurdwaras is to recite
the Guru Granth Sahib without an intervening interpretation. The Gurdwara is
the residence of the guru. The Sikhs identify themselves with the Guru Granth
Sahib, treating it as a living, speaking and engaging entity. It makes no sense
to others, but that is the essence of Indian plurality. It is the creative act of every
Sikh treating a text as a living person. The Sikhs do not see the theatre as a sacred territory where the rules of the
wider society can be suspended and offence created. But, brought up with
pluralism, they respect the Western institution of the apparent ‘free’ theatre. The Gurdwara as the ‘sacred’ However, they do not expect this historic western tension played as freedom to
colonise their own cultural space. They do not mock it nor humiliate it. Coexistence requires negotiation of mutual respect, not assuming a right to
offend the other’s sacred. But when the director of the theatre went beyond the
Western cultural sphere and dragged in the created sacred of another to humiliate
in the theatre’s safe territory, the theatre forfeited that respect and war was
declared. Like the colonist who used indigenous co-opted intermediaries to
legitimise their rule, the director justified his act by pointing out Bhatti’s origins
in a Sikh family. The Western theatre commands respect from people like the
Sikhs as long as it stays within its own boundaries. Once it crosses into the
territory of others then it has to respect the rules of the ‘other’, otherwise it can
only enforce its respect with the aid of the coercive power of the State to suppress
the people whose otherwise benign indifference is provoked into protecting
their own. The Sikhs stood outside the theatre for twelve days quietly protesting,
puzzled by the transgression of the director and the theatre’s production
manager’s refusal to engage in a dialogue. Dialogue is at the heart of Indic 121 Negotiating the Sacred II civilisation. The theatre production manager refused ‘dialogue’. The manager
set up a monologue a few days prior to the play being shown, and then, like the
imperialist, decided that the community had to learn ‘freedom of the theatre’. The director’s crusade, like a colonist, to ‘teach’ an ancient civilisation the West’s
newly acquired concept of freedom was an absurdity in itself. Like the French
who pursued culture evangelisation in East India in the seventeenth century,
only to be thrown out, like the missionaries who caused havoc with their
soul-saving crusades in ancient civilisations, the manager-director of the
Birmingham Repertory Theatre was on a mission, little realising the comedy of
the producer’s own position. The director’s freedom is that of the adolescent
emerging into puberty, full of bravado, threatening without the wisdom of
experience. The Sikh has inherited centuries of freedom with the sophistication
of critique and drama without crossing the limits of offence. The Gurdwara as the ‘sacred’ It is notable that the Sikh representatives who defended the Sikhs’ objections
to the play in public statements,45 made a clear distinction between the artha
and the abhinaya, stating that they had no problems with the substance of the
play, but objected to the depiction of the Gurdwara and symbolism of Guru
Granth Sahib, both of which are their sacred institutions separate from those of
others.46 In contrast, almost all articles and comments by leading figures from
the arts and media, including the petition by 700 leading luminaries of British
arts conflated everything into ‘freedom of speech’47 and ‘refusing debate’48 or
‘the right to challenge religion’!49 Most of the articles and statements attacking
Sikhs seemed to lack any rationale or analysis and depend more on histrionics. It is still not clear to Sikhs who this debate was supposed to be with, as the
audience was largely white middle class and some British educated Indians who
rarely if ever go to Gurdwaras. The community that this debate was supposed
to be initiated in was enraged by the disrespect shown to the Guru Granth Sahib
and the Gurudwara, the ‘Rasa’ had broken down. In context, Behzti brought two civilisations into contact but in unfortunate
circumstances. From the perspective of Indic civilisation, the West’s much
protected and prided freedom of expression extending to freedom to offend50
in arts and theatre51 is but an old conflict being played between those in power
and those without power that started in the Roman amphitheatre with its
captured audience in a walled confine. The Romans ridiculed the weak in the
bloody games in their amphitheatre. After them, the Church saw its right to be
free to offend human reason from the theatre of the pulpit. In the twentieth
century, human reason assumes a new born right to offend religious doctrine
from the theatre of arts. This has no relevance in Indic civilisation which is based
on the principle, ‘believe and let believe’. Freedom of conscience and expression
is a fundamental aspect of the non-dualist Indic civilisation. It allows critique
and dialogue but stops at gratuitous offence to preserve and nurture its essential 122 The monologue of liberalism and its imagination of the sacred in minority cultures plurality. The wider play in Behzti was the interaction between two different
civilisations with different historical evolution of ideas and political dynamics
of freedoms, and with different philosophical developments. The Gurdwara as the ‘sacred’ One which sees
the right to offend as its sacred duty, while the other which, in the words of
Yarrow, ‘needs to be recognised...for the subtlety and power of its grasp of the
nature and operation of individual and communal effects as embedded in its
performance tradition and in Natyasastra’.52 Endnotes
1 1 In a letter to the editor of the Guardian newspaper (UK) 23 December 2004, 700 of UK’s leading figures
from the arts world wrote: ‘We must defend freedom of expression’, and, ‘It is a legitimate function of
art to provoke debate and sometimes to express controversial ideas. A genuinely free, pluralist society
would celebrate this aspect of our culture’. Shelley King, David Edgar and 698 others: ‘We must defend
freedom of expression’, http://arts.guardian.co.uk/news/story/0,,1378818,00.html g
y
2 Nicholas Hytner, the artistic director of the National Theatre, told the BBC that theatre’s role was to
provoke powerful feelings: ‘The giving of offense, the causing of offense, is part of our business,’ he
said. Alan Cowell, ‘Rape, religion and artistic freedom’, International Herald Tribune, 29 December 2004. http://www.iht.com/articles/2004/12/28/features/bhatti.php
3 2 Nicholas Hytner, the artistic director of the National Theatre, told the BBC that theatre’s role was to
provoke powerful feelings: ‘The giving of offense, the causing of offense, is part of our business,’ he
said. Alan Cowell, ‘Rape, religion and artistic freedom’, International Herald Tribune, 29 December 2004. http://www.iht.com/articles/2004/12/28/features/bhatti.php 3 The Nyaya school of Hinduism is an analytic school which believes in the duality of mind and body. The Vaisesika school is atomistic. S. Radhakrishnan, 1989 (1978), Indian philosophy, vol. 2, New Delhi:
Oxford University Press. 3 The Nyaya school of Hinduism is an analytic school which believes in the duality of mind and body. The Vaisesika school is atomistic. S. Radhakrishnan, 1989 (1978), Indian philosophy, vol. 2, New Delhi:
Oxford University Press. 4 Edward Said, 1978, Orientalism, New York: Vintage Books. 5 5 Arvind Sharma (ed.) 2004, Advaita Vedanta: An Introduction, New Delhi: Motilal Benarsidas. 6 6 Adi Sankara was a Hindu monk who articulated the Advaita Vedanta (end of Vedas writings). In
modern times, Swami Vivekananda popularised Adi Sankara’s work in the nineteenth century. 7 The word ‘God’ may obscure its understanding. 8 8 In Jaap, Sahib, Dasaam Granth (the text of the tenth Sikh Guru). 9 9 Sri Guru Granth Sahib: (Avar na ko māranvārā.) ‘There is no other Destroyer than the Eternal,’ p. 391, line 2. 9 Sri Guru Granth Sahib: (Avar na ko māranvārā.) ‘There is no other Destroyer than the Eternal,’ p. 391, line 2. 10 The Sikhs have had ten human Gurus, one after the other from 1469 to 1708. They compiled their
teachings during their lives. Endnotes
1 This compilation in 1430 pages, mostly written in ragas, is called the Sri
Guru Granth Sahib and is revered as a living text, in the form of human by the Sikhs. The first eight
pages are the poetry of the first Guru, Guru Nanak, called the Jap, in which the Guru sets out the
philosophical context of Sikh teachings. From pages 14 to 1353 are the collection of hymns in 31 ragas. The last pages are the poetry of some main spiritual people from around India whose teachings were
consistent with those of the Sikh Gurus. 10 The Sikhs have had ten human Gurus, one after the other from 1469 to 1708. They compiled their
teachings during their lives. This compilation in 1430 pages, mostly written in ragas, is called the Sri
Guru Granth Sahib and is revered as a living text, in the form of human by the Sikhs. The first eight g
y
g
pages are the poetry of the first Guru, Guru Nanak, called the Jap, in which the Guru sets out the
philosophical context of Sikh teachings. From pages 14 to 1353 are the collection of hymns in 31 ragas. The last pages are the poetry of some main spiritual people from around India whose teachings were
consistent with those of the Sikh Gurus. 11 Sri Guru Granth Sahib: (Hukm hukam chalā rāhu.) ‘The order orders the paths to follow’. Jap, p. 2. 12 Sri Guru Granth Sahib: (Kiv kar Akha kiv salahi kio varni jana.) ‘How can we describe, how can we
l i
h
h
h
k
’ 11 Sri Guru Granth Sahib: (Hukm hukam chalā rāhu.) ‘The order orders the paths to follow’. Jap, p. 2. 12 12 Sri Guru Granth Sahib: (Kiv kar Akha kiv salahi kio varni jana.) ‘How can we describe, how can we
explain what happens, how can we know?’ Jap, p. 4. 13 Sri Guru Granth Sahib: (Nanak, akhan sabh ko akhai ik dui k siana.) ‘O Nanak, many speak of the
eternal truth to others, each one claims to be wiser than others’. (Vada sahib, badi nai, kita ja ka hove) ‘The great master with the most superlative names, ultimately
knows what has and will happen’. (Nanak, je ko apou janai, agai gaia na sohai.) ‘O Nanak, one who claims to know everything, shall not
be gifted with knowledge beyond his immediate world’. Jap, p. 5. Negotiating the Sacred II (Oh vekhai ona nadir na avai bahuta ehu vidan) ‘The eternal watches them, but they cannot see the
ultimate Source, that is the greatness of this mystery’. Jap, p. 7. Therefore, in Jap, Guru Nanak subtly asks the audience: Why go through gods who can do no better
than the human? 15 15 Chandra Bhan Gupta, 1954, The Indian Theatre, Munshiram Manoharlal Publishers: New Delhi, p. 107. 16 16 Bhagvat Gita 17 Radhakrishnan, op. cit., pp. 29-175. 18 Ibid., p. 248. 18 Ibid., p. 248. 19 Ibid., p. 430. 20 The four were, Apastamba, Gautama, Baudhayana and Vasistha, P. Olivelle, (trans.), 1999,
Dharmasutras, the Law Codes of Ancient India, Oxford: Oxford University Press, Contents and p. x 20 The four were, Apastamba, Gautama, Baudhayana and Vasistha, P. Olivelle, (trans.), 1999,
h
h
d
f
d
f
d
f
d
i
i
d The four were, Apastamba, Gautama, Baudhayana and Vasistha, P. Olivelle, (trans.), 1999,
Dharmasutras, the Law Codes of Ancient India, Oxford: Oxford University Press, Contents and p. xxv. 21 Manu was the lawgiver of ancient India who codified civil rules of behaviour and claimed divine
authority for these. It is considered that Manu became the nom de plume for a series of lawgivers. However, Manu did not claim universal imposition for these laws, and hence they had no meaning in
the world of sanyasis. 22 W. Doniger and B. Smith, 1991, The Laws of Manu, London: Penguin Classics, Penguin Books. 23 22 W. Doniger and B. Smith, 1991, The Laws of Manu, London: Penguin Classics, Penguin Books. 23 ‘In modern day language, Dharma is equated quite unfairly with religion’, Sri Krishna Kant, Vice
President of India, 1997, Convocation address, Sri Sathya Sai Institute of Higher Education, p. 3. http://www.sssu.edu.in/pdf/CG_Address1997.pdf 23 ‘In modern day language, Dharma is equated quite unfairly with religion’, Sri Krishna Kant, Vice
President of India, 1997, Convocation address, Sri Sathya Sai Institute of Higher Education, p. 3. http://www.sssu.edu.in/pdf/CG_Address1997.pdf 25 Teachings of A.C. Bhaktivedanta Swami Prabhupada, website owned by Bhaktivedanta Book Trust,
http://www.krishna.com/en/node/492 26 Kant, op. cit. 27 T.B. MaCauley, Minute dated 2 February 1835. Bureau of Education, 1965 (1920), Selections from
Educational Records, Part I (1781-1839), H. Sharp (ed.), Delhi: National Archives of India, pp. 107-117,
http://www.columbia.edu/itc/mealac/pritchett/00generallinks/macaulay/txt minute education 1835.html 27 T.B. MaCauley, Minute dated 2 February 1835. Bureau of Education, 1965 (1920), Selections from
Educational Records, Part I (1781-1839), H. Endnotes
1 14 14 Sri Guru Granth Sahib: (Eka Mai Jugat Viai tin chele parvan) ‘One source conceived the entire existence
and authorised the three’; (Ik sansari ik bhandari ik lae deban) ‘One the creator, one the sustainer and one the destroyer’ (talking
of Brahma, Vishnu and Siva); (Ik sansari ik bhandari ik lae deban) ‘One the creator, one the sustainer and one the destroyer’ (talking
of Brahma, Vishnu and Siva); (Jiv tis bhavai tivai chalavai jiv hovai furman) ‘Whatever is desired so will happen, they merely follow
the order’; (Jiv tis bhavai tivai chalavai jiv hovai furman) ‘Whatever is desired so will happen, they merely follow
the order’; 123 Negotiating the Sacred II Negotiating the Sacred II Sharp (ed.), Delhi: National Archives of India, pp. 107-117,
http://www.columbia.edu/itc/mealac/pritchett/00generallinks/macaulay/txt_minute_education_1835.html
28 David Ludden, 2006, Making India Hindu, Oxford: Oxford University Press, p. 21. 28 David Ludden, 2006, Making India Hindu, Oxford: Oxford University Press, p. 29 E.W. Marasinghe, 1989, The Sanskrit Theatre and Stagecraft, Delhi: Sri Satguru Publications (India
Book Centre). 30 Ibid., p. 185. 31 Ibid., p. 186. 32 Ibid., p. 186. p
33 Ibid., p. 197. p
34 R. Yarrow, 2001, India Theatre, Theatre of Origin, Theatre of Freedom, Richmond, Surrey UK:
.Routledge Curzon, p.118. 35 Chandra Bhan Gupta, op. cit., p.107. 36 ‘The Ankle Bracelet’,,Yarrow, op. cit., p. 113. Yarrow holds that tragic ends and realism is consistent
with Sanskrit theatre. 37 Bhatia Nandi, 1963, Acts of Authority/Acts of Resistance: Theatre and Politics in Colonial and Postcolonial 37 Bhatia Nandi, 1963, Acts of Authority/Acts of Resistance: Theatre and Politics in Colonial and Postcolonial
India, Michigan: University of Michigan Press, chapter 2. Bhatia Nandi, 1963, Acts of Authority/Acts of Resistan f
y/
f
India, Michigan: University of Michigan Press, chapter 2. 38 L.N. Rangarajan, 1992, Kautilya, The Arthashastra, New Delhi: Penguin Classics, Peguin Books. 39 Yarrow, op. cit., p.12. 40 Ibid., p. 27. 41 M. Hardt and A. Negri, 2001, Empire, Harvard: Harvard University Press. 42 41 M. Hardt and A. Negri, 2001, Empire, Harvard: Harvard University Press. 42 42 Yarrow, op. cit., p.12. Ibid., p. 13. Ibid., p. 13. 43 Ibid., p. 12. 43 Ibid., p. 12. 44 Sri Guru Granth Sahib, p. 138. 44 Sri Guru Granth Sahib, p. 138. p
45 ‘But Gurdial Singh Atwal, a Labour councilor and representative for the Council of Sikh Gurdwaras,
said: ‘It has caused a great hurt, and shows a lack of respect. The Sikh community had a small demand:
rather than setting it in a gurdwara, set it in a community centre.’ T. Branigan, 2004, ‘Tale of Rape at 124 The monologue of liberalism and its imagination of the sacred in minority cultures the temple sparks riot at theatre’, The Guardian, 20 December,
http://arts.guardian.co.uk/news/story/0,,1377285,00.html the temple sparks riot at theatre’, The Guardian, 20 December,
http://arts.guardian.co.uk/news/story/0,,1377285,00.html
46 46 ‘But many Sikh representatives argue that the issues have been misunderstood. Harmander Singh,
a spokesman for the advocacy group Sikhs in England, said concerns about the setting of the play had
gone unheeded for days before the violent protests. 52 Yarrow, op. cit., p. 21. 51 Nicholas Hytner, the artistic director of the National Theatre, told the BBC that the theatre’s role
was to provoke powerful feelings: ‘The giving of offense, the causing of offense, is part of our business’,
he said. Cowell, op. cit. Negotiating the Sacred II Sikh representatives had suggested that the play
would be far less offensive if the setting were changed from a temple to a community center, a proposal
the theater rejected.’ Cowell, op. cit. 47 Sir Christopher Frayling, chairman of the Arts Council said: ‘It sends out a message that there are
certain subjects about which they must never speak.’ Terry Kirby, 2004, ‘Violence and vandalism close
theatre’, The Independent, 21 December, http://www.independent.co.uk/arts/theatre/news/article25779.ece
48 Petition: ‘We must defend freedom of expression’. ‘It is a legitimate function of art to provoke debate
and sometimes to express controversial ideas. A genuinely free, pluralist society would celebrate this
aspect of our culture’. Letters, signed by 700 leading illuminaries in the arts: ‘We must defend freedom
of expression’, The Guardian, 23 December 2004, http://arts.guardian.co.uk/news/story/0,,1378818,00.html
9 49 Dominic Dromgoole, 2004, ‘Theatre’s role is to challenge religion’, The Guardian, 20 December 2004. http://arts.guardian.co.uk/features/story/0,,1377489,00.html 49 Dominic Dromgoole, 2004, ‘Theatre’s role is to challenge religion’, The Guardian, 20 December 2004. http://arts.guardian.co.uk/features/story/0,,1377489,00.html g
y
50 ‘”Artists” power to move brings with it a duty of intellectual integrity. Within the limits this duty
imposes, however, they must be free to offend — and this freedom, too, is sacred.’ Editorial, Times, 21
December 2004. 50 ‘”Artists” power to move brings with it a duty of intellectual integrity. Within the limits this duty
imposes, however, they must be free to offend — and this freedom, too, is sacred.’ Editorial, Times, 21
December 2004. 51 Nicholas Hytner, the artistic director of the National Theatre, told the BBC that the theatre’s role
was to provoke powerful feelings: ‘The giving of offense, the causing of offense, is part of our business’,
he said. Cowell, op. cit. 51 Nicholas Hytner, the artistic director of the National Theatre, told the BBC that the theatre’s role
was to provoke powerful feelings: ‘The giving of offense, the causing of offense, is part of our business’,
he said. Cowell, op. cit. 125 9
Blasphemy in a pluralistic society Jeremy Shearmur In this chapter, I will tentatively address a difficult issue: what we should make
of blasphemy today, in a society like Australia?1 My discussion falls into three
parts: considering blasphemy under the headings of truth, ridicule, and play. I
also discuss some different dimensions to the character of the offence. I conclude
with a discussion of some particular problems about blasphemy posed by the
fact that we are living in a pluralistic society, in the face of which I make some
specific — but inadequate — recommendations. I am happy to call them
‘inadequate’ just because my aim, at the end of the paper, is to open up what
seem to me some difficult problems rather than to resolve them. In my preparatory research, I came to the conclusion that the more that one
looks at some of the issues connected with blasphemy, say, at their history, the
more complex they turn out to be. The consequence is that if, as in the present
chapter, one is trying to speak in general terms, one becomes acutely aware of
just how thin the ice is upon which one is treading. As a result, it is bound to
be the case that, in addition to provoking the kind of critical reaction that I am
intending to provoke, and which I am hoping will lead to a fruitful response to
the issues which I am setting out to raise, I may also provoke all kinds of gnashing
of teeth on the part of those who have specialised knowledge on issues that I
deal with very briefly. All that I would say in my defence is that I am acutely
aware of my fallibility and, as a former student of Karl Popper’s, I am aware of
the need for criticism, which I look forward to receiving in due course. Let me start by saying a very little about a key problem. It is: What is the
character of blasphemy as an offence, and why was it thought appropriate to
punish it? My concern here will not be with its legal status,2 but with what the
underlying concerns have been. There seem to me to have been several lines of
argument. I will mention but five. 9
Blasphemy in a pluralistic society The first, as, say, we meet it for example in Leviticus 24, 16-18, is that the
punishment, even with death, of the blasphemer is simply seen as a command
of God, which is to be followed as such. The second is that blasphemy is understood as a substantive offence against
God. Here, humans who make accusations of it, and uphold the law in relation
to it are, as it were, acting in His defence, or in defence of His reputation. 127 Negotiating the Sacred II The third is slightly convoluted. It is that blasphemy is understood as
something which, if unpunished, will threaten those who do not punish it with
the judgement of God. Punishment of the blasphemer is, here, seen as a reaction
to the threat of the collective punishment by God of those who fail to punish
blasphemer. The fourth is the idea that an attack on religion is problematic because of
what are thought to be its social consequences, i.e., the destruction of civil order. This played a significant role during one period of British law, as is illustrated
by the following statement by Lord Chief Justice Hale: …to say, religion is a cheat, is to dissolve all those obligations whereby
the civil societies are preserved, and that Christianity is parcel of the
laws of England; and therefore to reproach the Christian religion is to
speak in subversion of the law.3 …to say, religion is a cheat, is to dissolve all those obligations whereby
the civil societies are preserved, and that Christianity is parcel of the
laws of England; and therefore to reproach the Christian religion is to
speak in subversion of the law.3 Finally, there is a more typically modern understanding. This is the notion
of blasphemy as an offence against the sensibilities of believers, and, further, as
something which because it so offends, is problematic because it is liable to lead
to a breach of the peace. Let me now move to the discussion of blasphemy, as such. I need briefly to
explain why I am discussing the issue of blasphemy under the three heads to
which I have referred: truth, ridicule and play.4 I do so because, historically,
one can note people as having been charged with blasphemy for activities that
fall conveniently under these somewhat different headings, and I wish to argue
that the issues that they raise are somewhat different, too. 9
Blasphemy in a pluralistic society Historically, one element in blasphemy has been a simple contesting of the
truth of various religious claims. This, for example, was one element in the
charges against Thomas Aikenhead at the end of the seventeenth century and
against G. W. Foote in the latter part of the nineteenth. In earlier times, people
who had wished to contest the truth of Christianity, were sometimes punished
— but for impiety. In the context of an orthodox Christianity feeling under
threat from Socinianism and Deism,5 Aikenhead’s youthful mixture of
contestation of and irreverent comment about Christianity was taken horribly
seriously, and he was executed.6 Foote, the editor of The Freethinker in the
1880s, combined denial of the truth of Christianity, and fierce argument to that
effect, with the use of ridicule.7 In the context of one of the trials of The
Freethinker, Lord Coleridge declared: I lay it down as law, that, if the decencies of controversy are observed,
even the fundamentals of religion may be attacked without a person
being guilty of blasphemous libel.8 128 Blasphemy in a pluralistic society This is generally accepted in Western countries today; but, for example, it may
be contentious for some Muslims. Foote, however, did much more than this. And that takes us to the second category, of ridicule. One of the features of The
Freethinker were various hostile cartoons about the life of Jesus, and more
generally illustrating in a satirical manner various themes from the Bible. The
latter included an unsubtly illustrated version of the verse from Exodus 33,
verses 22-3: ‘And it shall come to pass…that I will put thee in a clift of the
rock…and I will take away my hand and thou shalt see my back parts’.9 The
Freethinker’s cartoon, as the previous quotation from Lord Coleridge implicitly
indicated, raises the issue: Is the use of ridicule as part of an attack on religion
acceptable? Third, and of particular significance in our current context, is what I will
term the playful or aesthetic use of blasphemy. By this I mean blasphemy
committed not as part of a disagreement with, or an attack on religion (although
it is not clear that it would be used in such a manner by a devout religious
believer), but, instead, as used for literary or artistic effect. An obvious example
here is Salman Rushdie’s playing with themes from Islam to literary effect, in
his Satanic Verses. 9
Blasphemy in a pluralistic society We may here, similarly, consider the work of art, Piss Christ,
and also, say, in a much more minor key, parodies of the Creed, and so on,
produced by William Hone for political purposes in the early nineteenth
century.10 The key idea here is that religious material is played with, or used
for effect, in ways that believers could reasonably find offensive, but where the
motivation of those making use of this material is not to attack religion. With
this before us, let me now turn to the body of the paper, and to the issue of
truth. Truth Truth seems to me usefully looked at in terms of the kinds of consideration that
John Stuart Mill advanced in his On Liberty. I should state explicitly that my
concern, here, will be with the social consequences of epistemological fallibilism11
rather than with the specifics of Mill’s views; Jonathan Riley has argued
powerfully that Mill’s own arguments are best seen in the wider context of his
social philosophy, and that seen in that light, Mill is to be taken as arguing for
a more liberal view than that which is presented here.12 If it is being claimed
that various religious doctrines are true, then some kind of case has to be made
for them. What is involved here, may seem to get us into the territory of various
traditional arguments for the existence of God, such as the ‘cosmological
argument’. But what is actually offered in the context of religious proselatisation
may be rather different. Now if any such claims are being made as to the truth of religious doctrine,
we get straight into the issues which J. S. Mill discussed.13 He argued that all 129 Negotiating the Sacred II of our knowledge is fallible, and that if we are aiming for truth, our best course
of action is to submit our claims to open critical scrutiny. In this context, he was
then able to develop arguments against the suppression of opinion. To Mill’s
argument, it might be objected by the believer: ‘but our religious views are not
fallible’. However, those who agree with Mill can respond: ‘but you have
certainly not furnished us with telling arguments that show you views to be
anything but fallible.14 And your tendency, across history, to kill or to imprison
those who have objected to your claims, does not exactly speak for their strength
— for if the arguments were strong, why should they require support from the
hangman or the jailor’.15 It seems to me that, here, the case for allowing the public contestation of
religious doctrines is unanswerable. Not only is the Millian case a powerful one,
but since the Reformation we have lived in a society in which religious truth is
contested. Some limitations of the truth-based argument It is, however, worth pausing to consider what the intellectual issues to which
I have referred actually amount to. For this intellectual case has limitations; in
particular, it does not seem to me to offer the basis of a case for ‘freedom of
expression’. (Clearly, someone might say: ‘but the case for freedom of expression
does not depend on arguments like Mill’s, but, instead, relates somehow to ideas
about human agency, or is simply a “right”’. But it is just not clear on what basis
such a right could be established if, say, it involves a purported entitlement to
behave in offensive ways towards others.) Let me return to the fallibilistic argument from Mill. As far as I can see, all
that is required for the fulfilment of the kinds of requirement that Mill set out,
is that there be some place where such contestation can take place, and that the
results of debates about such matters be freely available in the society.19 What
this means, is that there need to be journals in which such matters can be
discussed, and places where people interested in such things can meet. This is
a very different matter from some entitlement to bombard other people with
challenges to their views when they do not wish to encounter them,20 or, say,
a right even to demonstrate as opposed simply to the advancing of arguments. More generally, an awareness of human fallibility seems to me in principle
compatible with restrictions on the dissemination of information which has not
been exposed to some appropriate form of inter-subjective scrutiny. This might,
however, today be accomplished without the suppression of opinion, by way
of the requirement that certain kinds of material be published only in a setting
in which an indication can be given of the basis on which others disagree with
what is being advanced. For example, rather than the making of claims about,
say, holocaust revisionism illegal — which may itself backfire21 — there could
be a legal requirement that the material be posted only to web pages that allow
for responses by scholars who disagree with the case being argued. Let me also offer some further considerations against the taking of Millian
arguments in an expansive way. There is much that goes to constitute our
day-to-day lives that might quite fairly be called necessary fictions. Truth If there are Catholics and Protestants — to say nothing of Muslims
and Jews, and also non-believers — who are members of a single society, then
clearly there is disagreement about religious truth.16 Indeed, the very beliefs
of these groups implicitly or sometimes even explicitly involve the view that
the other religious beliefs are incorrect. If we are living in a society that is
pluralistic in this sense, the notion that the discussion and the contestation of
religious ideas should be suppressed would seem an absurdity. (It is, however,
worth noting that Jewish criticism of Christian claims, particularly its claims
about itself as a supposed fulfilment of the Old Testament, was frequently
suppressed.17 ) Given that all the groups to whom I have referred —
contemporary Judaism apart — are universalistic and attempt to convert others,
this is surely an invitation to contestation.18 Today, Christendom accepts the case for openness of argument. (Although
one wonders sometimes what the more dogmatic believers would do, if they
could get their hands on the reins of enough political power.) It is not clear that
this case for openness of argument is accepted by all Muslim clerics. Attitudes
there, at least in terms of the views of some clerics, seem closer to those that
were common in Western countries several centuries ago, although there are,
of course, also liberal Muslims. There is also room for debate as to just what is
supposed to be accorded what kind of protection, and it is quite another matter
what anyone actually becomes concerned about (the Rushdie case had strong
political elements to it). At least in principle, to voice disagreement about some
key issues is problematic, even if all that is involved is intellectual engagement. Of course, in practical terms, no Muslim is going to be able legally to take action
against Christian ministers who may say things within their religious
congregations that are at odds with Islam. Further, Muslims have, I think, to
face the fact that if they choose to live in a society like Australia, they are living
in a society in which the open intellectual contestation of their beliefs — whether 130 Blasphemy in a pluralistic society by sceptics, or by those who favour other religions — will take place. Truth Further,
if such contestation is conducted along the ‘decent’ lines that Lord Coleridge
suggested, other citizens will expect that those engaged in such contestation
will, if necessary, receive full protection from the law. Some limitations of the truth-based argument We typically
have a view of the world, and of ourselves, which we implicitly take to be correct
and live by, and which we are not interested in having challenged. Most people,
if they are not suffering from depression, are moral heroes in their own internal
narratives; and all of us operate within a framework of assumptions — many of
which will not, in fact, be true — which we certainly do not wish to have
challenged, unless we choose to open them up to such challenges. Some of those 131 Negotiating the Sacred II reading this will, like myself, be academics, and this means that certain things
that we do will be subject to fairly constant challenge and criticism. More
generally, the cultures in which academics live and by which our ideas are
broadly informed are subject to certain kinds of challenge and scrutiny. But
typically — and, I think, quite properly — the ideas that constitute the
assumptions under which we live on a day-to-day basis, are not usually submitted
to such scrutiny. In part, this is because one can, as it were, hardly live in the
open — with all aspects of ourselves and our conduct exposed to the critical
scrutiny of others all the time. In part, it is because many of these ideas are
simply things that we use while getting on with other, more pressing, matters. At the same time, one of the features of life in Australia which I have come to
value — as compared, say, to the USA — is that there is a degree of accountability
here to one another, in our ordinary culture, for what we believe.22 Now, the ideas on the basis of which we live may well be incorrect or
misguided. Sometimes, we may become annoyed with other people because of
what they believe and act on. But broadly speaking, it seems to me that we are
not entitled to challenge them (unless we are invited to do so), unless it is the
case that those ideas and their actions (or failures to act) impinge significantly
upon us, or upon others with whose well-being we have a legitimate concern. In this respect, while intellectual contestation is important, there is no case for
the view that one is simply entitled to harangue other people, unasked, about
their ideas or view of the world. Some limitations of the truth-based argument You should recognise 132 Blasphemy in a pluralistic society that not only may people (understandably) start to avoid you, but you may
legitimately expect them to make you aware of the range of objections to your
views, if they are not too polite to do so. This is a somewhat difficult matter. For consider, say, the Protestant who
thinks that if you are not saved, you will suffer in Hell.24 How, such a person
might say, can they be expected to conduct the ordinary niceties of life with
you, without drawing this to your attention — and repeatedly, because so much
hinges on it? They would surely do this, if you were in physical danger; and
how much more important is your fate in Eternity? But just as that person may
feel that he or she cannot engage in polite social chit-chat with those on the edge
of a precipice, so the objects of their concern may find that they really cannot
take constantly being engaged with on this subject. The only courses of action
may be to agree not to raise the issue, or to separate. There is, however, also another aspect to all this. It is that, as academics who
are used to the cut and thrust of discussion, we typically come to know what
we are letting ourselves in for if we enter into it. For each of us, there will be
certain kinds of standards to which we know that we will be held accountable
in our various professions and the sub-areas of these that we inhabit. I can, by
now, make a pretty fair judgement as to what the (different) standards are, to
which different groups of philosophers, of political scientists, of historians of
economic thought, and so on, will hold me accountable if I give a paper to them. Further, if I make some claim — say, about the arts — in a gathering of people
who deal with the arts professionally, while I cannot judge quite how what I
say will be appraised, I will have at least a rough idea about the kinds of things
that might be thrown at me. Such knowledge, however, is typically acquired by us gradually through
our lives, and sometimes somewhat painfully. Some limitations of the truth-based argument (Or rather, there is a sense in which one may
do this, in respect of people who are friends of ours, provided that they have
the option to stop being our friends if they find this wearing!) Such ideas —
consider, say, whether or not they think that eating meat is wrong — in effect
become like the tastes of these other people: things that we simply live with
provided that they neither positively harm others in ways that we find
intolerable, nor make us really uncomfortable. If they harm others,23 we may
feel it our duty to open up a discussion about them. If they simply make us
uncomfortable, we may need to modify how we interact with these people (for
example, just moaning about them to friends who agree with us), or possibly
even break off contact with them. Now, the ideas on the basis of which we live may well be incorrect or
misguided. Sometimes, we may become annoyed with other people because of
what they believe and act on. But broadly speaking, it seems to me that we are
not entitled to challenge them (unless we are invited to do so), unless it is the
case that those ideas and their actions (or failures to act) impinge significantly
upon us, or upon others with whose well-being we have a legitimate concern. This may seem a somewhat timid viewpoint, and you may even object to it. But if you do, just ask those who know you, how many of your views about
things they think to be incorrect, and ask yourself: ‘would I like to be constantly
open to contestation about them?’ Alternatively, you may yourself be committed
to some form of proselytisation; for example, for some religious view, or for
some form of environmentalism. If this is the case, it is worth bearing in mind
just to what extent you may be infringing upon the kinds of informal rules that
we normally live by concerning such matters, and thus pausing to consider just
how obnoxious you may be. If you are going to do this, however, it seems to
me that what is good for the goose is good for the gander. Some limitations of the truth-based argument I can still recall coming to
university, as a gauche young man who simply did not know how these things
worked, and how painful the first few steps were. I have also had the (at times
difficult) experience of operating within different disciplinary contexts. By
contrast with all this, we may forget that other people may not have this kind
of experience behind them. As, say, we are sometimes reminded when a capable
but academically inexperienced mature student joins an introductory course in
the subject that we teach, people may have specific views which they believe
to be true, but where their asserting them as true does not mean quite what it
would normally in academic contexts. If I make claims that I assert to be true,
this opens me up as an Aunt Sally, for everyone to have a go at. If people do not
have an academic background, they may not realise what they are getting into
if they make similar claims in an academic context. But, on the other hand, unless
we are anthropologists or sociologists with an interest in these issues, we are 133 Negotiating the Sacred II not likely to have any real knowledge as to how their claims about the status of
such views are made and negotiated in different areas of even our own society. I would, in this kind of situation, suggest that we should exercise something
the name of which I hardly dare speak, namely, paternalism. That is to say, it
seems to me that, unless it is clear that the people in question wish to have their
ideas contested as we would have our claims contested, their views be treated
more like the expression of tastes. This will typically not be fully adequate to
the cognitive character of those views, and there is a risk that we may be insulting
to them. But perhaps better this, than the exposure of their ideas to the full force
of the critical apparatus that we, given our training, may be able to bring to
bear upon them. This, while fully legitimate in its place, may be inappropriate
if used in a contest in which people are not accustomed to it, or do not realise
what they are getting themselves into. I am reminded of a conversation that a
good friend of mine reported that he had with his partner. Some limitations of the truth-based argument They had some
disagreement about their personal affairs, and he said to her: ‘We should talk
this out.’ She said: ‘No way — you, as a philosophy lecturer, are a specialist in
arguments. I would not have a chance, even if I was right about everything.’ Just as in this case, it seems to me that in the kind of situations about which
I have been talking, respect for other people as persons may mean that we should
treat their views as having a somewhat different status from what they may
present them as having. After all, we may be quite used to discovering that ideas
which we have cherished, and have worked on for a while, turn out just to be
no good. Given our professional training, we typically have the resources to
cope with such things, even though the experience may not be pleasant. However, it may not be something that other people, without these resources,
can easily cope with at all. It is certainly not clear to me why coping with it
should be thrust upon them, without their indicating positively that they wish
to get into such a situation. Accordingly, there seems to me a case for the protection of some ideas from
criticism — although it is not one which compromises what I might call the
minimalist Millian case for there being a public sphere in which any idea may
be contested (including, of course, ideas as to what such a sphere should be like). At the same time, it is important to note that the case for the moderation of
criticism was made in the face of the vulnerability of the people whom I was
discussing, and also under the assumption that what they were doing was not
having adverse effects on others. The (interesting) social task here would be to
create ways of behaving which protect the vulnerable, but which do not shelter
the powerful. 134 Blasphemy in a pluralistic society From intellectual engagement to ridicule So far, I have been dealing with issues of truth and falsity, and of the
contestation, back and forth, of particular claims. But what of ridicule? Is it ever
appropriate to ridicule or to insult things about which other people have deep
feelings? Here, it seems to me, we get onto more difficult ground. For if, say, a Christian
believes that God, the all-powerful creator of the universe, gave his Son not only
to suffer the humiliations and pangs of mortal life, but to die a gruesome death
for the sake of each of us — then to poke fun at this, seems simply grotesque. It certainly is not something that can rest upon a J. S. Mill-style defence. May
anything be said in its favour? Two interesting lines of argument here were
advanced by the English Freethinker G. W. Foote, who ended up being
imprisoned for blasphemy in the 1880s. First, consider those who are not believers. They may find the hegemony of
the ideas to which they are constantly and oppressively exposed, simply
intolerable. I recall, here, a comment made by a distinguished sociologist of
education, who was Jewish, and who undertook an ethnographic exercise, over
a period of three years, in a fundamentalist Baptist school. He later recalled that,
he hit a problem: …in one of the phases of research, the last phase, the write-up. Obviously, my ox had been gored. As a non-Christian at Bethany I
experienced repeated proselytising by students, parents, teachers, and
the superintendent of the school. I felt that I had been assaulted by the
most arrogant people I had ever met in my entire life…Obviously, all of
this had gotten to me in more ways tha[n] I understood; it was coming
out in my writing, though I didn’t want it to. I did not want chapter
one, let alone chapter two, three, four, five or six, to be redolent of the
anger suggested by terms such as ‘assault’ and ‘arrogance’.25 …in one of the phases of research, the last phase, the write-up. Obviously, my ox had been gored. As a non-Christian at Bethany I
experienced repeated proselytising by students, parents, teachers, and
the superintendent of the school. From intellectual engagement to ridicule I felt that I had been assaulted by the
most arrogant people I had ever met in my entire life…Obviously, all of
this had gotten to me in more ways tha[n] I understood; it was coming
out in my writing, though I didn’t want it to. I did not want chapter
one, let alone chapter two, three, four, five or six, to be redolent of the
anger suggested by terms such as ‘assault’ and ‘arrogance’.25 In this case, the sociologist could get away from the group which he had studied. But what if one were a member of a minority group, who were endlessly the
targets of something similar from the wider culture? In this case, the sociologist could get away from the group which he had studied. But what if one were a member of a minority group, who were endlessly the
targets of something similar from the wider culture? In this setting, at the very least, it would be fully understandable to let off
steam, to other members of one’s group, about the kind of things to which one
was subjected. This, indeed, was pretty much Foote’s explanation for one of the
things that his newspaper did: for running his cartoons, poking fun at passages
from the Bible. His argument was that the material was sold in a newspaper for
Freethinkers; it was not forced upon religious believers, and it would be pretty
clear to the believer what the character of the journal was, should they come
across a copy. 135 Negotiating the Sacred II Foote, however, offered another line of argument, too. Let me present it, in
his own words: Christians may, of course, urge that their feelings on such a subject as
religion are sacred, and a few superstitious Freethinkers may concede
this monstrous position. I do not. The feelings of a Christian about Father, Christians may, of course, urge that their feelings on such a subject as
religion are sacred, and a few superstitious Freethinkers may concede
this monstrous position. I do not. The feelings of a Christian about Father,
Son and Holy Ghost, are no more sacred than my feelings on any other
subject. I have no quarrel with persons, and I recognise how many are
hurt by satire. But the world is not to be regulated by their feelings, and
much as I respect them, I have a greater respect for truth. From intellectual engagement to ridicule Every mental
weapon is valid against mental error. And as ridicule has been found the
most potent weapon of religious enfranchisement, we are bound to use
it against the wretched superstitions which cumber the path of progress. Intellectually, it is as absurd to give quarter as it is absurd to expect it.26 I do not know what you might make of this. In some ways, for Foote to take this
line in a situation where he might well go to jail for it, and indeed did, was
courageous. At the same time, it is also clear that such satire need not be on the
side of what is right, even though it may be horribly effective. Consider, in this
context, the use by the Nazis, and by the regime in the Soviet Union, of art and
representations which dehumanised those with whom they disagreed. Not only
were those — often very powerful — representations exercised in the interests
of views which were, surely, wrong, but they also served to dehumanise people,
in ways which opened them to what became ghastly treatment. All told, there
seems to me something to be said for treating people with decency, and respecting
their sensibilities, even in cases where we think that they are deeply wrong. Such concerns, however, look to me pressing only in the case of ordinary
people, and those who do not exercise power. The powerful can, surely, well
look after themselves — although even there, there is surely something to be
said for recognising the human in everybody. If we accept this argument, then
there is also reason to accord them a certain kind of respect — which would
include not submitting those things about which they have the strongest and
most tender feelings, to the full force of our abuse. Play, or the aesthetic use of blasphemy So far, our concern has been with those who wish to take issue with the content
of religion. But if we consider blasphemy in contemporary contexts, this seems
to me not, typically, what we are concerned with. Instead, what has been thought
problematic, has been the use of religious ideas for aesthetic reasons or, to put
it bluntly, a certain kind of playing around with them. Consider Salman Rushdie’s Satanic Verses. Here, he made a certain amount
of mileage, for his own aesthetic purposes, with themes from Islam: the so-called
‘satanic verses’ episode itself, which related to the supposed interpolation — 136 Blasphemy in a pluralistic society and then deletion when their character as illegitimate interpolations was revealed
— of some verses into the text of the Koran, which legitimated the role of some
local gods as intermediaries between people and God. In addition, Rushdie played
around with the idea of the denizens of an imaginary brothel, who boosted their
custom by naming themselves after the wives of the Prophet. Was Rushdie’s
aim to attack Islam, in something like the way in which Foote did Christianity? Surely not; it was more a matter of his using these ideas, as instruments in his
novel.27 Similarly, one might say, the artist responsible for Piss Christ was
playing around with Christian symbolism. What is to be said about this? Clearly, there are legitimate uses of religious
material, not only in a devotional context, but — because of the extent to which
our culture and language is (still) permeated with them — for the purpose of
playfulness and parody. Two things, however, are worth noting. First, such
usage is parasitic upon the public, cultural significance of the material. The
reason why a form of words that draws upon well-known passages from the
Bible, or from the Creed, or upon religious symbolism, may make the impression
that it does, is because of its resonance with the material from which it is drawn,
and the kinds of public associations that it has. Second, there is the problem of
the offence that it may cause. It might be said: ‘No offence is intended — the artist had his or her own
private intention.’ This, however, will not wash, because of the public
significance of the material. Play, or the aesthetic use of blasphemy After all, the aesthetic weight of the material is
drawing precisely upon its public, in this case religious, meaning.28 What is
more, people have to take into account the public meaning of what they are
saying. If, say, someone gets up and yells ‘Fire, Fire’ in a crowded, darkened
cinema, it is possible that they may have had their own particular intentions —
for example, they might be giving an impromptu performance of some free verse. But this is no excuse; unless there is a very distinctive preparation of the people
who will be receiving the material,29 such that this serves to diffuse its normal
social meaning, it is clear how it will be interpreted. And the poet will justly
suffer the consequences of their action, if they don’t do so. In the course of the discussion of the Salman Rushdie case, John Updike is
reported as having said: ‘it is perhaps in the nature of modern art to be
offensive…In this century if we are not willing to risk giving offence, we have
no claim to the title of artists’.30 But bear in mind what it is to be offensive.31
First, if someone is offensive to you, they may surely risk retaliation, and if they
thought about it, they might well realise that this is what they should expect. Why, after all, should the rest of us be expected to sit there and have people
behave offensively towards us, without reacting to it? This, of course, then leads
to that element in blasphemy cases in which people are taken to have committed
an offence, because their conduct is liable to lead to a breach of the peace. 137 Negotiating the Sacred II Second, being offensive — especially to the defenceless — may simply be a
horrible thing to do. Once, say, people understand just what kind of offence
certain kinds of conduct, or the improper use of Aboriginal designs, may give
to the people in question, it is surely inexcusable to persist in giving offence. It
is a horrible action, and also made worse by the sheer lack of power of the people
on whom it is inflicted. However, it is not at all clear, say, that the kinds of
offence that may be given to ordinary Muslims, or to devout Catholics or to
conservative-minded evangelical Christians is any less. Play, or the aesthetic use of blasphemy Just think, in the latter
cases, of how they might experience what is taking place. As I mentioned before,
in their view, the creator of the universe chose to humble himself, take on human
form and to suffer, so as to bring about the salvation of all who would accept
this gift…and this is then mocked, turned into a travesty, or used as a prop for
the purposes of comedy. I am not advocating the jailing of the people who might
behave in this way. But it seems to me that we should think twice about the
idea that such activities can easily be defended as part of a supposed right to
self-expression. However, should we go along with such ideas we face a problem;
one that I am not sure how to resolve. Blasphemy in a pluralistic society But what
about Jehovah’s Witnesses, the Mormons, and New Agers, to say nothing of
Little Pebble?32 for this purpose. Think about this in an Australian context: Christianity — in
all its forms? Islam — bearing in mind that, for Shiites, the twelfth (occluded)
Imam would also require protection? Judaism? Aboriginal beliefs? But what
about Jehovah’s Witnesses, the Mormons, and New Agers, to say nothing of
Little Pebble?32 There is also a further problem here: namely, just what the content of that
which receives such protection will be. It is often — and surely rightly — said
that, say, Aboriginal cultures are not static, but evolving. The same, surely,
might be the case for what others hold to be Holy. But this poses a problem: if
what is so recognised acquires special protected status, it can surely become the
subject of politics. In an early episode in the Garfield cartoon series, Garfield, a
fat striped cat, was made to say something like: ‘One good thing about being a
cat is, any food you touch, becomes yours.’ The Holy, or what plays a key role
in your culture such that it demands protection, might equally become the
vehicle for a not dissimilar kind of acquisitive opportunism. In addition, as I noted earlier, one feature of religious pluralism is that the
key tenets of many religions commit their devotees to denying the claims of the
devotees of other religions. It is just not clear that there can be respect and
mutual tolerance, if you take the claim to the truth of your own beliefs seriously,
and it is integral to your own views that the other people have got things terribly
wrong. Finally, here, although I can address this question only very briefly, there
is an interesting problem concerning whether, with regard to these matters,
there is an area between the private and the public.33 I earlier referred to one
line of argument in Foote, in which he argued that the cartoons in the Freethinker
were produced for an audience of secularists, and that this should have been
clear from the character of the publication. It is possible that the same might be
argued, in respect of the Gay News blasphemous libel case. Blasphemy in a pluralistic society When Salman Rushdie wrote, he knew full well what he was doing. The
knowledge that he displayed — in playing around with the ‘satanic verses’
episode and even in his use of the names of the wives of the Prophet — required
some real knowledge of Islam. But it should equally have been clear to him that
what he was doing would have been found highly offensive by some Muslims. Clearly, there was much more to the episode than this. There was a strong
political element to the attention that his work received, and Muslims could find
much other material that would give offence, but which was not singled out for
condemnation in this way. Many of his Western readers, however, might have
found these aspects of his work unusual, and while — especially if they came
across some commentary upon them — they might have been able to understand
them, they would surely not have experienced the kind of emotional reaction
that some Muslims may have felt. One obvious problem however, is that now that we are a pluralistic society:
in Australia, there are many faiths and sensibilities, and even societies which
are relatively homogenous are affected by what is done elsewhere. Just what
protections are needed, and what should be protected? In the face of the Rushdie case, one reaction in the UK was that some Muslims
there wished for Rushdie to be had up for blasphemy. They swiftly discovered,
however, that in Britain laws against blasphemy related only to the Christian
religion — and even, it might appear, to the doctrines of the Church of England. There was, then, a call for the widening of blasphemy law, so that it would apply
to other religions. This, however, poses a problem; namely, what should count 138 Blasphemy in a pluralistic society for this purpose. Think about this in an Australian context: Christianity — in
all its forms? Islam — bearing in mind that, for Shiites, the twelfth (occluded)
Imam would also require protection? Judaism? Aboriginal beliefs? But what
about Jehovah’s Witnesses, the Mormons, and New Agers, to say nothing of
Little Pebble?32 for this purpose. Think about this in an Australian context: Christianity — in
all its forms? Islam — bearing in mind that, for Shiites, the twelfth (occluded)
Imam would also require protection? Judaism? Aboriginal beliefs? Blasphemy in a pluralistic society Here, a short poem
by James Kirkup, depicted a Roman centurion expressing his love for Jesus, by
means of various forms of sexual activity conducted with his just-dead body.34
It would seem as if the poem was intended to express a genuine devotion, but
its public meaning was highly offensive, and Mrs Mary Whitehouse, of the
British National Viewers and Listeners Association — a crusader for morality
— brought a private prosecution against it for blasphemous libel, which was
successful.35 One might ask, however: but should not the fact that it appeared
in Gay News indicate that it was produced for, as it were, a niche market, such
that those who might find this offensive would understand that it was not for
them? But in this particular case, the reaction of their own readers, and that of
those working on the paper towards the poem, suggested that this defence might
not be open to them,36 even if it could be defended in a wider context. Finally, here, although I can address this question only very briefly, there
is an interesting problem concerning whether, with regard to these matters,
there is an area between the private and the public.33 I earlier referred to one
line of argument in Foote, in which he argued that the cartoons in the Freethinker
were produced for an audience of secularists, and that this should have been
clear from the character of the publication. It is possible that the same might be
argued, in respect of the Gay News blasphemous libel case. Here, a short poem
by James Kirkup, depicted a Roman centurion expressing his love for Jesus, by
means of various forms of sexual activity conducted with his just-dead body.34 139 Negotiating the Sacred II Where does all this leave us? I am initially inclined to say that, in a pluralistic
setting, there is good reason for ditching the crime of blasphemy. At the same
time, there is surely a case for prohibiting by law offensive behaviour that is
liable to provoke a breach of the peace. At another level, it would seem to me
that it is reasonable to regard as morally offensive the production of things that
would deeply offend harmless people: consider, again, the deliberate violation
of things that Aboriginal people hold as sacred, but now generalise this to the
core concerns of others. 4 There is also of course the issue of blasphemy as expletive; but this I will not discuss here. For a rather
different classificatory approach, see Anthony Fisher and Hayden Ramsay, ‘Of Art and Blasphemy’,
2002, Ethical theory and moral practice, vol. 3, pp. 137-67. Blasphemy in a pluralistic society At the very least, this should mean that we don’t fete
work which gives such offence, or express solidarity with it if it is criticised. This does not mean that we should silently acquiesce to calls for the death or
imprisonment of those who produce it. I said, above, that I am inclined to offer that response. But there are problems
about it, not least this question of what, and to what extent, things get protection,
and who gets to decide. It is surely only core concerns that should receive such
protection. But who gets to decide what people’s core concerns are? And does
the protection that might be accorded to the core concerns of major religions,
really extend to, say, the fantasies of some group of ratbags who thinks that
aliens have kidnapped them a space ship, and then returned them to Earth with
remarkable news for mankind? I am left unsure about what we should do concerning these matters. One
might say: be respectful of work that has a public meaning as sacred, in the
cultures within which our work is produced and in which we would expect it
to be disseminated. And over and above this, be mindful of the sensibilities of
the vulnerable. But such a response is clearly inadequate, and I am happy to
leave this problem to others, to see what they make of it. Andres Serrano, when his Piss Christ was attacked, commented that he
thought that its destruction was an act of ‘desecration’.37 Clearly, I might be
tempted to take any criticism of my paper in similar terms — if I were not, in
these matters, influenced by Karl Popper, who stressed fallibilism and the
importance of holding everything open to criticism, and who argued that it is
good for us, even if we don’t enjoy it. 2 On which, see Reid Mortensen, 1994, ‘Blasphemy in a secular state: A Pardonable sin?’, UNSW Law
Journal, vol. 17, no. 2, pp. 409-31. A useful brief overview of blasphemy laws in different states and
territories is provided at: http://www.caslon.com.au/blasphemyprofile2.htm. 3 Quoted by Mortensen, ibid., p. 411.
4 1 I would like to thank Geoff Stokes for conversation and for some most useful comments on a draft of
this paper, from some of which I was not able to benefit on this occasion, for reasons of space, and also
to David Wall and the editors for some useful criticism and suggestions. 1 I would like to thank Geoff Stokes for conversation and for some most useful comments on a draft of
this paper, from some of which I was not able to benefit on this occasion, for reasons of space, and also
to David Wall and the editors for some useful criticism and suggestions.
2 On which, see Reid Mortensen, 1994, ‘Blasphemy in a secular state: A Pardonable sin?’, UNSW Law
Journal, vol. 17, no. 2, pp. 409-31. A useful brief overview of blasphemy laws in different states and
territories is provided at: http://www.caslon.com.au/blasphemyprofile2.htm. Endnotes
1 Foote is
discussed at length in Joss Marsh, 1998, Word Crimes, Chicago & London: University of Chicago Press. 8 Q
t d i M
h ibid
160 g
f
p
y
g
g
internet at: http://homepages.ihug.co.nz/~freethought/foote/blasphemy/0bcontents.htm. Foote is
discussed at length in Joss Marsh, 1998, Word Crimes, Chicago & London: University of Chicago Press. 8 Quoted in Marsh, ibid., p. 160. 8 Quoted in Marsh, ibid., p. 160. 9 It is reproduced in Marsh’s Word Crimes, ibid., p. 142. 10 For some discussion of which, see Marsh, ibid. 11 11 That is to say, that aspect of Mill’s views about toleration, that stems from the stress that he placed
on the idea that human knowledge was fallible, that currently accepted views may not be correct, and
so on. 11 That is to say, that aspect of Mill’s views about toleration, that stems from the stress that he placed
on the idea that human knowledge was fallible, that currently accepted views may not be correct, and
so on. 12 See Jonathan Riley, 2005, ‘J. S. Mill’s Doctrine of Freedom of Expression’, Utilitas, vol. 17, no. 2,
pp. 147-79. 13 It is striking from the discussion in Hunter and Wootton, that at times the production of apologias
for the truth of Christianity against imagined opponents gave rise to people taking the imagined
opponents’ case seriously and finding it telling. 14 For critical discussion of claims to infallible knowledge in the Shi’ite tradition of Islam, compare the
work of Abdolkarim Soroush, 2002. Some of his essays are available in English in his Reason, Freedom
and Democracy in Islam, Mahmoud Sadri and Ahmed Sadri (eds), London etc: Oxford University Press. 15 Clearly, an argument might be made for the suppression of material on the grounds that it might
mislead the immature. But it is not clear that, for example, the history of the suppression of heresy or
of Jewish objections to Christianity, can really be placed in that category. j
y
y
p
g
y
16 This oversimplifies, in that there was, historically, the recognition of a kind of pluralism within
Islam, as in the milet system of the Ottoman Empire (cf. 16 This oversimplifies, in that there was, historically, the recognition of a kind of pluralism within
Islam, as in the milet system of the Ottoman Empire (cf. Endnotes
1 1 I would like to thank Geoff Stokes for conversation and for some most useful comments on a draft of
this paper, from some of which I was not able to benefit on this occasion, for reasons of space, and also
to David Wall and the editors for some useful criticism and suggestions. 140 Blasphemy in a pluralistic society 5 Socinianism, named after Lelio Francesco Maria Sozzini (1525–1562), was a sixteenth and seventeenth
century religious movement, which called into question the deity of Christ, and emphasised, by contrast,
the unity of God. It is sometimes regarded as an early form of unitarianism. Deism, a loose movement
influential in the seventeenth and eighteenth centuries, while accepting the existence of God and his
role as a creator, rejected revealed religion in favour of a sceptical and rationalistic approach, and also
rejected the idea that God subsequently intervened in his creation. 6 See, notably, Michael Hunter, 1992, ‘“Aikenhead the Atheist”: the Context and Consequences of
Articulate Irreligion in the Late Seventeenth Century’, in Michael Hunter and David Wootton (eds.)
Atheism from the Reformation to the Enlightenment, Oxford: Clarendon Press; and also W. Cobbett, T. B. Howell et. al. (eds), 1819, A Complete Collection of State Trials, xiii, London, pp. 917-40. For
background, see also the other articles in the Hunter and Wootton collection, and also J. A. I. Champion,
1992, The Pillars of Priestcraft Shaken, Cambridge: Cambridge University Press. 6 See, notably, Michael Hunter, 1992, ‘“Aikenhead the Atheist”: the Context and Consequences of
Articulate Irreligion in the Late Seventeenth Century’, in Michael Hunter and David Wootton (eds.)
Atheism from the Reformation to the Enlightenment, Oxford: Clarendon Press; and also W. Cobbett, T. B. Howell et. al. (eds), 1819, A Complete Collection of State Trials, xiii, London, pp. 917-40. For
background, see also the other articles in the Hunter and Wootton collection, and also J. A. I. Champion,
1992, The Pillars of Priestcraft Shaken, Cambridge: Cambridge University Press. 7 Foote’s fascinating Prisoner for Blasphemy, London: Progressive Publishing, 1886, is available on the
internet at: http://homepages.ihug.co.nz/~freethought/foote/blasphemy/0bcontents.htm. Foote is
di
d t l
th i J
M
h 1998 W d C i
Chi
& L
d
U i
it
f Chi
P 7 Foote’s fascinating Prisoner for Blasphemy, London: Progressive Publishing, 1886, is available on the
internet at: http://homepages.ihug.co.nz/~freethought/foote/blasphemy/0bcontents.htm. Endnotes
1 y
(
http://www.eurozine.com/articles/2004-03-25-ivanov-en.html), and also in Christendom at such times
as Jews were not being persecuted. In addition, in the period immediately after the Reformation, a
system was adopted in which the religion of a country was simply settled by the choice of the ruler. 1 http://www.eurozine.com/articles/2004-03-25-ivanov-en.html), and also in Christendom at such times
as Jews were not being persecuted. In addition, in the period immediately after the Reformation, a
system was adopted in which the religion of a country was simply settled by the choice of the ruler. 17 17 Cf, for example, Richard Popkin ‘Jewish Anti-Christian Arguments as a Source of Irreligion from
the Seventeenth to the Early Nineteenth Century’, in Hunter and Wootton (eds) Atheism from the
Reformation to the Enlightenment. Isaac Troki’s Faith Strengthened, for example, something that it is
tempting to say should be compulsory reading for all evangelical Christians, is currently difficult to
obtain in printed form (although a version is now available on the internet at: 17 Cf, for example, Richard Popkin ‘Jewish Anti-Christian Arguments as a Source of Irreligion from
the Seventeenth to the Early Nineteenth Century’, in Hunter and Wootton (eds) Atheism from the
Reformation to the Enlightenment. Isaac Troki’s Faith Strengthened, for example, something that it is
tempting to say should be compulsory reading for all evangelical Christians, is currently difficult to
obtain in printed form (although a version is now available on the internet at: http://faithstrengthened.org/). It is striking that Hans Joachim Schoeps was able to remark in the
‘Foreword to the Second Edition’ of his The Jewish-Christian Argument: ‘This book made its first
appearance in 1937, behind closed doors, as it were, for the authorities at that time in power in Germany
allowed its sale only in Jewish bookstores and to Jews. Then, when it was noticed that it was a
“dangerous” book, it was prohibited completely.’ See his The Jewish-Christian Argument, London:
Faber, 1965, p. xiv. See also Richard Popkin, 2006, Disputing Christianity: The 400-year-old Debate over
Rabbi Isaac Ben Abraham Troki's Classic Argument, Amherst, N.Y.: Humanity Books. http://faithstrengthened.org/). It is striking that Hans Joachim Schoeps was able to remark in the
‘Foreword to the Second Edition’ of his The Jewish-Christian Argument: ‘This book made its first
appearance in 1937, behind closed doors, as it were, for the authorities at that time in power in Germany
allowed its sale only in Jewish bookstores and to Jews. Endnotes
1 27 The ideas of which Rushdie made use were familiar enough, and had not been thought of as
particularly contentious, in discussions of Islam. See, on this, and on many of the other complexities
of the Rushdie case, the discussion in Michael M.J. Fischer and Mehdi Abedi, 1990, Debating Muslims:
Cultural dialogues in postmodernity and tradition, Madison, Wis: University of Wisconsin Press. 28 I have in mind here something along the lines of the view developed by Quentin Gibson and used
by him and others in the context of textual interpretation in the history of political thought; cf. James
Tully (ed ) 1988 Meaning and Context Princeton: Princeton University Press 28 I have in mind here something along the lines of the view developed by Quentin Gibson and used
by him and others in the context of textual interpretation in the history of political thought; cf. James
Tully (ed.) 1988, Meaning and Context, Princeton: Princeton University Press. Tully (ed.) 1988, Meaning and Context, Princeton: Princeton University Press. 29 Or, say, unless they were a recently arrived visitor to the country who could quite reasonably not
be expected to appreciate the significance of what they were saying. 29 Or, say, unless they were a recently arrived visitor to the country who could quite reasonably not
be expected to appreciate the significance of what they were saying. 30 Quoted in Daniel Pipes, 1990, The Rushdie Affair, New York: Birch Lane Press/ Carol Publishing
Group, p. 108. 30 Quoted in Daniel Pipes, 1990, The Rushdie Affair, New York: Birch Lane Press/ Carol Publishing
Group, p. 108. 31 31 Of course, what is offensive is something which will change over time and will also be
context-dependent, which introduces further problems that I cannot even attempt to deal with, given
the space at my disposal. 32 ‘Little Pebble’ is the name of the leader of a Catholic religious group, who in 2005 was in court to
answer charges relating to sexual conduct towards an under-age member of his flock. For a site which
gives some indication of the charges against him, see http://users.bigpond.net.au/wanglese/pebble.htm
(Viewed January 3 2006); for the group’s web page see: http://www shoal net au/~mwoa/ (Viewed 32 ‘Little Pebble’ is the name of the leader of a Catholic religious group, who in 2005 was in court to
answer charges relating to sexual conduct towards an under-age member of his flock. Endnotes
1 22 The tradition of freedom of religion in the USA seems to be taken as a freedom to live without ones
religious ideas being challenged and this — or so it seemed to me when I was living there — extended
to beliefs about many other matters, in a manner that contrasts with day-to-day life in Britain or Australia. 23 Clearly, what counts as another may be contentious — say, with regard to vegetarianism, or, say,
the treatment of some socially unpopular minority — but on the face of it there is a difference between
how something is to be appropriately handled if it is broadly socially contentious in the society in
question, as compared to ones having what is very much a minority viewpoint in the society in question. (For example, pleas for vegetarianism in a society of meat eaters need to be made in journals rather than
at the dinner table.) Even issues that are contentious may need to be treated in a circumspect manner:
if, say, the vegetarian chooses to eat with meat eaters in a society in which there were lively divisions
and debate about vegetarianism, it would hardly be appropriate for them to accompany the meal with
a running commentary about sentience, slaughterhouses, and quotations from Peter Singer. n this, Joseph Alleine, Alarm to unconverted sinners etc, London: Nevil Simmons, 1993 etc. 25 Alan Peshkin, 1992, ‘Experience Subjectivity’, Qualitative Interest Group, College of Education,
University of Georgia, 1992 conference proceedings, Keynote Address. http://www.coe.uga.edu/quig/proceedings/Quig92_Proceedings/peshkin.92.html (Viewed January 21,
2005) 25 Alan Peshkin, 1992, ‘Experience Subjectivity’, Qualitative Interest Group, College of Education,
University of Georgia, 1992 conference proceedings, Keynote Address. 25 Alan Peshkin, 1992, ‘Experience Subjectivity’, Qualitative Interest Group, College of Education,
University of Georgia, 1992 conference proceedings, Keynote Address. http://www coe uga edu/quig/proceedings/Quig92 Proceedings/peshkin 92 html (Viewed January 21 y
g
g
y
http://www.coe.uga.edu/quig/proceedings/Quig92_Proceedings/peshkin.92.html (Viewed January 21,
2005). 26 Quoted from Foote, Prisoner for Blasphemy (see note 2). 27 26 Quoted from Foote, Prisoner for Blasphemy (see note 2). 27 27 The ideas of which Rushdie made use were familiar enough, and had not been thought of as
particularly contentious, in discussions of Islam. See, on this, and on many of the other complexities
of the Rushdie case, the discussion in Michael M.J. Fischer and Mehdi Abedi, 1990, Debating Muslims:
Cultural dialogues in postmodernity and tradition, Madison, Wis: University of Wisconsin Press. Endnotes
1 Then, when it was noticed that it was a
“dangerous” book, it was prohibited completely.’ See his The Jewish-Christian Argument, London:
Faber, 1965, p. xiv. See also Richard Popkin, 2006, Disputing Christianity: The 400-year-old Debate over
Rabbi Isaac Ben Abraham Troki's Classic Argument, Amherst, N.Y.: Humanity Books. 18 At the very least, if one is offering arguments that are supposed to convince others, this opens up
the possibility of contestation; for example, to the effect that the arguments are not valid or are not
telling. 19 18 At the very least, if one is offering arguments that are supposed to convince others, this opens up
the possibility of contestation; for example, to the effect that the arguments are not valid or are not
telling. 19 19 I am offering this as an argument as to what follows from epistemological fallibilism, not as an
interpretation of Mill. There has been argument about just what follows from Mill’s own arguments,
with some (for example, John Skorupski, 1989, John Stuart Mill, London: Routledge) taking the line
that I am commending here, while others (for example, Jonathan Riley in his ‘J. S. Mill’s Doctrine of 19 I am offering this as an argument as to what follows from epistemological fallibilism, not as an
interpretation of Mill. There has been argument about just what follows from Mill’s own arguments,
with some (for example, John Skorupski, 1989, John Stuart Mill, London: Routledge) taking the line
that I am commending here, while others (for example, Jonathan Riley in his ‘J. S. Mill’s Doctrine of 141 Negotiating the Sacred II Negotiating the Sacred II Freedom of Expression’) argue that Mill’s own argument leads to a defence of more expressive activiti
such as demonstrations. Freedom of Expression’) argue that Mill’s own argument leads to a defence of more expressive activities,
such as demonstrations. 20 But where, obviously, if they offer arguments or try to convince others that they are right, they
deserve everything that they get in response. 20 But where, obviously, if they offer arguments or try to convince others that they are right, they
deserve everything that they get in response. 20 But where, obviously, if they offer arguments or try to convince others that they are right, they
deserve everything that they get in response. 21 Compare, in this context the controversy in 2004-05 opened up by the Iranian President Mahmoud
Ahmadinejad’s remarks about the Holocaust. Endnotes
1 It is striking that the Pakistani academic Ijaz Hussain, in
an opinion piece ‘COMMENT: Who is more civilised: Iran or the West?’ was able to make what read
like a reasonable case for Ahmadinejad’s views, by way of describing the legal suppression of dissent
about the Holocaust in several countries, and providing a list of people or organisations which had
questioned it. He did not, say, refer to works which contest such claims, such as Deborah Lipstadt’s
Denying the Holocaust (New York: Free Press, 1993), or to the various publications on the David Irving
libel trial against Lipstadt in the UK. Ijaz Hussain, 2006, ‘COMMENT: Who is more civilised: Iran or
the West?’ Daily Times, Pakistan, January 4 2006; see
http://www dailytimes com pk/default asp?page=2006%5C01%5C04%5Cstory 4-1-2006 pg3 5) 21 Compare, in this context the controversy in 2004-05 opened up by the Iranian President Mahmoud
Ahmadinejad’s remarks about the Holocaust. It is striking that the Pakistani academic Ijaz Hussain, in
an opinion piece ‘COMMENT: Who is more civilised: Iran or the West?’ was able to make what read
like a reasonable case for Ahmadinejad’s views, by way of describing the legal suppression of dissent
about the Holocaust in several countries, and providing a list of people or organisations which had
questioned it. He did not, say, refer to works which contest such claims, such as Deborah Lipstadt’s
Denying the Holocaust (New York: Free Press, 1993), or to the various publications on the David Irving
libel trial against Lipstadt in the UK. Ijaz Hussain, 2006, ‘COMMENT: Who is more civilised: Iran or
the West?’ Daily Times, Pakistan, January 4 2006; see
htt
//
d il ti
k/d f
lt
? 2006%5C01%5C04%5C t
4 1 2006
3 5) 22 The tradition of freedom of religion in the USA seems to be taken as a freedom to live without ones
religious ideas being challenged and this — or so it seemed to me when I was living there — extended
to beliefs about many other matters, in a manner that contrasts with day-to-day life in Britain or Australia. 36 It is striking, however, that from Rictor Norton’s personal account of the background to the
publication http://www.galha.org/glh/214/norton.html, not only was there considerable protest from
within the readership of the journal, from people who found it offensive, but Norton indicates that
they ‘did not relish defending a poem that we frankly realized was rather sick’, even if it was sincere.
37 See ‘Andres Serrano talks with Judith Ahern’, Photofile 53, April 1998, pp. 8-13; see p. 13. Serrano
claimed that he saw it as a desecration ‘not toward the mage but toward Christ himself’. But this seems
to me simply an indication of the degree to which — not unlike the poet in the cinema — his personal
interpretation was of kilter with the public meaning of what he had produced. 36 It is striking, however, that from Rictor Norton’s personal account of the background to the
publication http://www.galha.org/glh/214/norton.html, not only was there considerable protest from
within the readership of the journal, from people who found it offensive, but Norton indicates that
they ‘did not relish defending a poem that we frankly realized was rather sick’, even if it was sincere.
37 See ‘Andres Serrano talks with Judith Ahern’, Photofile 53, April 1998, pp. 8-13; see p. 13. Serrano
claimed that he saw it as a desecration ‘not toward the mage but toward Christ himself’ But this seems 36 It is striking, however, that from Rictor Norton’s personal account of the background to the
publication http://www.galha.org/glh/214/norton.html, not only was there considerable protest from
within the readership of the journal, from people who found it offensive, but Norton indicates that
they ‘did not relish defending a poem that we frankly realized was rather sick’, even if it was sincere. Blasphemy in a pluralistic society y
g
y
37 See ‘Andres Serrano talks with Judith Ahern’, Photofile 53, April 1998, pp. 8-13; see p. 13. Serrano
claimed that he saw it as a desecration ‘not toward the mage but toward Christ himself’. But this seems
to me simply an indication of the degree to which — not unlike the poet in the cinema — his personal
interpretation was of kilter with the public meaning of what he had produced. Section IV The argument in this section considers artistic creation and self-expression
achieved within restriction. It is hard to see that the value claimed for artistic
expression, of showing the world anew, or of expressing the common concerns
of humanity, requires that art should be unrestricted. While it might be assumed
that artists must work without restriction in order to express themselves, ‘fetters’
do not necessarily constrain what may be represented. For instance, films that
respect modesty may also express sexual longing and consummation. Restriction
is a condition for creativity, and is a framework within which creativity is
recognised. As artist and curator, Christopher Braddock coordinated the exhibition Votive:
sacred & ecstatic bodies including the works of Ian Breakwell and Cathy de
Monchaux (Britian), Pierre & Gilles (France), and Megan Jenkinson (New Zealand). The exhibition responded to controversy surrounding Tania Kovat’s exhibit
Virgin in a Condom at the Museum of New Zealand Te Papa Tongarewa in 1998
and the work of Andres Serrano, Braddock writes about issues of blasphemy in
artworks that engage with collisions between sacred imagery and the body. This
chapter addresses complex relationships in the artworks between, on the one
hand, attitudes of devotion and, on the other, severe critique of the Church. The
artists exhibiting in Votive transgress, but avoid obvious attacks through an
emphasis on religious ecstasy and sensory qualities of religious symbols, rather
than their meaning. Braddock argues that the gallery provides a context for the
exploration of devotional forms excluded from the Church. The next two chapters explore externally, and internally imposed censorship. Post revolution Iranian cinema has been subject to heavy censorship particularly
in the manner in which the romantic relationships between men and women are
represented on screen and the language that is used in the script — un-Islamic,
profane, blasphemous or sacrilegious utterances, acts and gestures are forbidden. In a culture that upholds religious norms stringently and where idolatry and
imitating the creative act of God is considered to be highly sacrilegious, Michelle
Langford is surprised that no official sanction has been imposed against producing
films but instead, cinema is used as a vehicle for promoting the State’s agenda
of educating people about Islamic values. While the lack of ‘fundamentalism’
may be surprising, it is clear that the state still seeks to control the social construct
of the pure and the impure within these films. Endnotes
1 For a site which
gives some indication of the charges against him, see http://users.bigpond.net.au/wanglese/pebble.htm
(Viewed January 3, 2006); for the group’s web page, see: http://www.shoal.net.au/~mwoa/ (Viewed
January 3, 2006). 33 I would particularly like to thank Geoff Stokes for discussion on this point. 34 33 I would particularly like to thank Geoff Stokes for discussion on this point. 34 34 The poem is reproduced at http://gaytoday.badpuppy.com/garchive/events/070502ev.htm (Viewed
November 3, 2005). 35 See, for a brief report,
http://news.bbc.co.uk/onthisday/hi/dates/stories/july/11/newsid_2499000/2499721.stm, which includes
a picture of Mrs Whitehouse. (Viewed November 3, 2005.) 142 143 Section IV However, Langford’s analysis of
Iraqi cinema shows that even stringent controls do not necessarily undermine
an artist’s ability to express themselves, or to critique the State. 145 Negotiating the Sacred II Many Australian Aboriginal societies utilise secrecy as a fundamental norm
for regulating speech. In Yolngu society, a person requires ‘the right’ to speak
on a topic, and the anthropologist Ian Keen has reported that this secrecy is
supported by self censorship, even when a person knows information, they may
refuse to acknowledge that they have this knowledge. Silence presents a means
of negotiating the sacred and preserving the sanctity of Indigenous knowledge. In Indigenous Australia, collective experiences of a community are sacred and
cannot be recounted. Stories are the only way by which such a sacred world
may be represented, but stories embody ancestral beings and places and are
sacred in their own right so they should not be needlessly evoked. In our final chapter, Caroline Josephs, a writer, makes a very personal
exploration of this system. In identifying with the Yolngu and giving credibility
to their religious accounts of the land, she interprets the silence surrounding
such stories not as censorship but as respect, or awe, in the face of the sacred,
and as a means of negotiating the relationship she has with others. By employing
various forms of storytelling — braiding genres of personal, cultural, conceptual
storytelling — the author covers three aspects of silence in relation to one
particular story — the Wagilag Sisters Story. Josephs weaves what she cannot
say about the Yolngu Wagilag Sisters story into her own story of ‘coming to
know’ the story and the place of their dance. 146 Locating ‘blasphemy’ in the polemical The beginning of the twenty-first century has brought into focus issues of
right-wing religious fundamentalism (be it Islamic or Christian) even more
sharply. The dangers and dilemmas of essentialist religious paradigms have
joined a list of vitally important global issues to be reckoned with, arguably as
important as global poverty and global warming. It is in this light that
bearing-witness to blasphemy becomes part of a debate about tolerance within
a pluralistic society. As Kyla McFarlane described in the catalogue for the exhibition Votive: Sacred
and ecstatic bodies, 2 in the last decade, Australia and New Zealand bore witness
to artwork seemingly crossing the boundaries that constitute blasphemy: In October 1997, Melbourne’s National Gallery of Victoria closed an
exhibition of works by American artist Andres Serrano after two physical
attacks by members of the public on his photograph Piss Christ (1987). This incident followed earlier claims by Christian groups and senators
in the USA that the work was indecent and obscene.3 Serrano had
received funding from the National Endowment for the Arts and,
following a complaint from the American Family Association, Senator
Jesse Helms proposed that indecent or offensive works should not
continue to receive funding. New York Senator Alphonse D’Amato also
condemned the work in Congress. His outrage, and that of Helms, was
publicly echoed by many constituents.4 Serrano’s work is a photographic enlargement of a small commercially produced
crucifix immersed in Serrano’s own urine. Without the title of the work to
indicate the substance of piss used to create the image, the viewer might revel
in the sublimely romantic connotations of a misty atmospheric sky at sunset
striking at the heart of the crucifixion story with its hope of resurrection after
death. But Serrano’s titling ensures that the viewer has to confront the dilemma
that this is a vat of urine. A severing of the unity of state and church has reinforced the freedoms of
the individual to roam across boundaries making a question of ownership of
religious symbols and their employment difficult to answer. The Catholic
Archbishop of Melbourne, Dr. George Pell, who considered Piss Christ
blasphemous, applied unsuccessfully for a Supreme Court injunction to prevent
the National Gallery of Victoria from exhibiting the work. 10
Blasphemy and the art of the political
and devotional
Christopher Braddock Christopher Braddock A breakdown in the unity of state and church has altered the context in which
blasphemy might be understood. Rather than viewing blasphemous libel as
intrinsically linked through the Ecclesiastical courts within the unity of State
and Church, the emphasis has shifted to the individual in society whose freedom
of artistic expression is constrained instead by the secular laws of defamation
and obscenity. In this light, artists such as Andres Serrano and Tania Kovats
embrace a freedom to use dramatic and visually confronting binary oppositions
in their works of art.1 Andres Serrano juxtaposed the crucifix with urine, and
Tania Kovats, a statuette of the Virgin Mary with a condom. Such iconography
provokes the possibility that these artworks might function as simultaneously
political and devotional: powerfully critical of church institutions, while
aesthetically operating as symbols that evoke reverence. In this context it is
significant that, in the debates that followed the controversies surrounding these
works of art, the arts establishment emphasised the division between church
and state in justification of the artists’ right to freedom of expression, and the
difference between temple and museum was highlighted in discussion about the
contextual setting of the museum as a place in which 'blasphemy' could not
occur. Artistic intention, and the context in which these works are witnessed,
are debated as factors in determining the possibility of blasphemy within the
divide of state and church, museum and temple. In this chapter I will argue that interpretations of graphic Medieval images
of the body that avoid binary opposition are useful in a reappraisal of the
controversy surrounding artworks such as Serrano’s Piss Christ and Kovat’s
Virgin in a Condom. This raises questions about the role of devotional images. Do they exist primarily to reinforce the strict teachings of the church or can
they be used to examine those beliefs? If we accept the latter as a possibility,
could these confrontational images signify divine revelation in the body, the
very essence of the Incarnation? 147 Negotiating the Sacred II Locating ‘blasphemy’ in the polemical Justice Harper cast
doubt on the need for the common law of blasphemous libel in Victoria, saying
that Australia ‘need not bother with [it]’ because contemporary Australia is a
pluralist, tolerant society.5 Justice Harper distinguished the Victorian position
from the English law stating that blasphemous libel is ‘an anachronism of English
history from a time when the State was intrinsically linked with the Church, 148 Blasphemy and the art of the political and devotional through the Ecclesiastical courts, and the unity of State and Church was not
transported to the Australian Colony. This position is supported by the fact that
Victoria does not recognise an established Church under section 116 of the
Australian Constitution’.6 Now that the state and the church are no longer
inseparable, the question of authority and how religious iconography is employed
appears to shift to wider concerns of freedom and artistic expression as
constrained by the laws of defamation and obscenity. A calculation of the level
of offence experienced by the viewer moved to centre-stage. McFarlane points to a similar reaction of public outrage with respect to Tania
Kovat’s Virgin in a Condom: In a matter of days, Tania Kovats’ Virgin in a Condom (1992) was stolen
by a visitor to the Museum of Contemporary Art in Sydney. Exhibited
as part of ‘Pictura Britannica’, a show of work by young British artists,
it provoked numerous letters of protest and complaints from incensed
Christians and Muslims when it toured to Te Papa/Museum of New
Zealand the following year.7 Virgin in a Condom is a strikingly polemical work. On the one hand, the seven
and a half centimetres tall mass-produced statuette is a religious object made for
the purpose of evoking the Virgin Mary’s divine intercession and has been
covered with an object used, more or less exclusively, for covering erect penises
or a variety of sex toys, such as ‘butt-plugs’. Clearly, many viewers understood
the work as presenting an analogy to the Virgin Mary as a dildo. As I will argue,
a reappraisal of these binary interpretations might suggest that, on the other
hand, the statuette is religiously shrouded or concealed, visually resembling a
work such as Piero della Francesca’s Madonna della Misericordia (circa 1445) ,
in which the Madonna protects the people with her mantle. Locating ‘blasphemy’ in the polemical This condom as
shroud evokes notions of the polarised role models for women within Catholicism:
that women are unfairly torn between virgin and whore in the archetypal models
of Mary, the Mother of God, and Mary Magdalene, the redeemed whore. As
well as this, the sculpture strategically provokes discussion about the Catholic
Church’s prohibition of contraception by the use of condoms. Neither of these
issues, by the way, had any prominence whatsoever in the New Zealand media
coverage surrounding the controversy over Virgin in a Condom’s presence in Te
Papa. The lack of media coverage on these key issues highlights an assumption
that artists intend to shock rather than engage in public debate. Such an
assumption overlooks the possibility that artist’s may intend to provoke public
discussion on these issues, or to explore unconventional views on religion within
a museum context. I will return to this point in discussing the role of the museum
as providing a space for unconventional views of religion below. The perceived polemical nature of the work contributed to Virgin in a Condom
becoming, arguably, the most publicised work of art in the history of New 149 Negotiating the Sacred II Zealand, Tania Kovats would not reply to any invitation to participate in the
exhibition Votive: Sacred and ecstatic bodies that I curated in Wellington in 2002. Her Australasian experience, which included receiving abusive mail and death
threats, had been so traumatic that she wanted nothing more to do with us. Her
representative, the London-based Asprey Jacque Gallery, indicated that the
direction of her work was changing to the extent that another inclusion in a
religious project would misrepresent the artist. Arguments surrounding the accountability of public expenditure in public
institutions such as galleries form strong undercurrents to these debates about
the control and use of religious symbols. In contrast with the National Gallery
of Victoria, Te Papa Tongerewa Museum of New Zealand stood their ground. They refused to remove Virgin in a Condom from the Pictura Britannica exhibition
and backed their right to be an artistic conscience of society. In the words of
the concept curator, Ian Wedde, the museum had to be a free space for several
kinds of expression. It had to be available ‘for the expression of divergent and
controversial views’.8 Some of these controversial views may be unorthodox
religious or theological perspectives. Locating ‘blasphemy’ in the polemical Serrano contends that Piss Christ stems from his religious upbringing, and
that the work upholds his religious conviction that the body can be seen as a
means for obtaining religious redemption. As Serrano puts it: The photograph, and the title itself, are ambiguously provocative but
certainly not blasphemous…My Catholic upbringing informs this work
which helps me to redefine and personalise my relationship with God. My use of such body fluids as blood and urine in this context is parallel
to Catholicism’s obsession with ‘the body and blood of Christ’. It is
precisely in the exploration and juxtaposition of the symbols from which
Christianity draws its strength.9 Who then, should decide whether something is blasphemous? The church or
religious group who claims ownership and control, or the artists themselves? One standard model for the interpretation of art involves consideration of
the artist’s intentions. Within a context of the divide between state and church
and the concomitant freedom of the individual, such a model seems apt. Our
knowing that Serrano sees himself as working within the parameters of his own
faith seems to alter our immediate impression that this is a case of blasphemy. It is as though, given his Christian upbringing, Serrano feels entitled to use the
religious images as he pleases, and that, as a Christian, he was incapable of
blaspheming. This idea of membership giving entitlement to use images however
you like raises questions about how ‘membership’ is determined, and whether
there can be levels of membership and entitlement. 150 Blasphemy and the art of the political and devotiona A conflicting position about how we should interpret a work of art appeals
to the conventional meaning of the symbols. If we read the image at face value,
it is still his piss, a human by-product of waste not normally associated with the
reverence of God. The media recently condemned American soldiers for the
blasphemy of urinating on the Koran during the military campaign in Iraq. If
this is clearly an act of blasphemy in relation to what is held sacred to Muslims,
and was correctly interpreted as an attack on the sentiments of the Muslim
prisoners they had captured, we would not expect a Christian to engage in a
similar act with respect to their own faith. Locating ‘blasphemy’ in the polemical This raises the question of what role
the gallery plays in providing a context for interpreting a work of art, and its
appreciation. The context of witnessing blasphemy Is viewing Piss Christ in an art gallery different from contemplating or
worshipping the image in a church? According to an institutional theory of art,
the identification of something as art influences the manner in which something
is interpreted. Such a theory of art is proposed by George Dickie where a ‘work
of art’ must satisfy the dual conditions of an artefact made by ‘a person who
participates with understanding in the making of an artwork’ together with a
confirmation of its status as an artwork ‘by some person acting on behalf of the
Artworld’.10 According to Dickie both conditions are necessary to confirm the
status of a work of art. From this perspective, does the institutional context of
the museum reserve a place for artistic critique that is separate from those
contexts within which we plausibly interpret something to be blasphemous? In
other words, does it reserve a space that permits artists to manoeuvre across
parameters of critique and devotion? This is a significant question as it has
implications for the role of the gallery as providing a space for alternative views
of religion and for the practice of religion. Serrano’s work can be compared to Ian Breakwell’s Deep Faith (2001), which
was included in the Votive exhibition. In Deep Faith, a slide projection features
an X-ray of the throat of a woman who had swallowed a small crucifix which
had lodged in her throat. Breakwell found the X-ray while he was an
artist-in-residence at the Cardiff Anatomy Unit. The gradual and repeated
dissolving of a second slide over the first featuring the words ‘deep faith’, a
reference to the 1972 pornographic film Deep Throat, sexualises the work’s
imagery. This overlay of images hovers between acceptance of Christian doctrine,
indeed an obsessive consumption of the Body of Christ, and the sexual act of
fellatio. This reference plays on devotional allusions, toying with the notion of
complete acceptance of Christian doctrine, while aligning this with a sexual act. The image also has a violent subtext, with the figure becoming an intrusive
lodgement in the throat. As McFarlane comments: 151 Negotiating the Sacred II This association of the figure of Christ with the interior of the body also
echoes Serrano’s submersion of a similar figure in his own urine in Piss
Christ. The context of witnessing blasphemy In both works, the body of Christ is strongly associated with the
functioning of the human body, at the beginning and end of digestive
process. In the former, the figure is taken into the throat and remains
uncomfortably lodged there, whilst in the latter, it is associated with
expulsion of waste.11 Ian Breakwell, Deep Faith, 2001, copyright The Estate of Ian Breakwell
M di t d b
h t
hi
d
t d
ithi th
d Pi Ian Breakwell, Deep Faith, 2001, copyright The Estate of Ian Breakwell Mediated by photographic processes and presented within the museum, do Piss
Christ and Deep Faith operate outside of the realm of devotion, and only as a
critique of the teachings of the Church? Do they, in other words, simply critique
from the sidelines? Mediated by photographic processes and presented within the museum, do Piss
Christ and Deep Faith operate outside of the realm of devotion, and only as a
critique of the teachings of the Church? Do they, in other words, simply critique
from the sidelines? As an artist, the question of where these artworks are witnessed was on my
mind when my work was included in an exhibition in New York City entitled
The Divine Body.12 The venues for the exhibition covered an array of institutional
settings. The gallery at Columbia, the Cathedral of St John the Divine, and a
theological seminary: art, worship and pedagogy. The distribution of the art
works seemed to have been dictated by economic imperative. Those of higher
economic value, such as the Andres Serrano and the Eric Fischl, were provided
with the security of the Columbia’s gallery, while other works were spread out
across the other venues. It occurred to me to ask: ‘What would have happened
if Piss Christ had been exhibited in St John the Divine?’ There might be people 152 Blasphemy and the art of the political and devotional who think Piss Christ is not blasphemous in a museum, but blasphemous in a
place of worship. To the best of my knowledge the work has never been exhibited
in a place of worship. But, would this be blasphemous? What would happen if
my cibachrome photographs of naked tattooed models were exhibited in St John
the Divine? These works explore white-trash inspired tattoos inscribed close to
nipple, buttock and pubis, intimately photographed. The context of witnessing blasphemy The works aim to provoke
a collision between the practice of tattoo outlawed within most Judaeo-Christian
circles and an edification of the body. And the question of placing Piss Christ in a church struck at the heart of a
dilemma in viewing both Piss Christ and Virgin in a Condom. On the one hand,
I see them as overly simplistic slogan-like artworks that are blasphemous in their
collision of clearly defined sacred and profane imagery; I would not photograph
the Koran floating in a pool of urine. And, on the other hand, my acceptance of
an institutional theory of art inclines me to think of the context of the art gallery
as a site of questioning and critique, not a site of devotional practice. Yet,
Breakwell’s aestheticisation of the image — projected in a dimly lit space and
viewed against the organ music of the late French composer Olivier Messian —
clearly point to an atmosphere of reverence if not of devotion. A conclusion that works of art accused of being blasphemous are so simply
because they break religious dogma, or are exhibited in a sacred space, is
problematic. The conclusion provokes questions about the role of religious
symbols in ecclesiastical settings, and suggests that they exist primarily to
reinforce the strict teachings of the church. A religious painting in a church
could also examine or tease-out the nature of those beliefs. My earlier suggestion
that the museum might reserve a place of artistic critique outside the realms of
worship seems now too simplistic, and undermines the possibility of ‘religious’
art operating across the parameters of critique and devotion. Religious law and sacred bodies Ian Breakwell’s video, The Sermon, 1984, which was also exhibited in the Votive
exhibition, subtly explored interpretations of ‘the law’ in Catholic teaching and
its relationship with the desires of the body. In so doing, Breakwell shifts our
focus from a cleric reinforcing the law of the church to an individual overcome
by a sense of blasphemy in how he occupies his own body. Kyla McFarlane
writes in Votive: The Sermon is…[a] sardonic commentary on the effects of the law as
stated by the institutional voice, it takes the format of a television sermon
in a book-lined study reminiscent of the set of Stars on Sunday. The
speaker, a seated priest, begins by calmly reminding the viewer that we
have erred and offended against ‘the laws’ and that clarification is needed
regarding areas of guidance. As the monologue progresses, however, his 153 Negotiating the Sacred II avid pleasure in addressing the perverse and sinful becomes apparent,
with the tone of the address moving from sage, fatherly advice to a
frenzied tirade against the lusts of the flesh and original sin. The sermon
is subject to a kind of leakage, as a perverse desire meets with the rigidity
of the institutional voice…Breakwell’s spoken text is littered with
references to bodily fissures and seepages, such as ‘the rupturing of
membranes’ during birth, or the leaking of sperm from a condom
perforated with holes so that the marriage act can still be performed
within the guidelines of the faith whilst collecting semen for laboratory
tests. Or, finally, the gutting of the stomach of the depraved. Absurd
and viscous, The Sermon ironically describes the impress of the body
and its filthy excesses on the intellectual rigour of the law. In attempting
to contain and restrain, the law itself becomes perverse and obsessive. By the video’s end, desire and the law become blurred to the point of
misrecognition.13 In the video, bodily function, and our relationship with the body, is inscribed
with the categories of the pure and the impure. The body is thought of as a kind
of temple, which requires us to treat it in a certain way. Yet the attempt to
achieve purity is undermined by the body itself; its own secretions defile it. Religious law and sacred bodies Julia Kristeva argues that categories of abomination such as food taboos or
corporeal alterations, topologically correspond to one’s being able to have access
to a place — the holy place of the Temple. Impurity, therefore, has a relationship
to what happens outside the Temple. But this relationship is not contingent on
a physical existence of the Temple: for Judaism, the Temple’s function remained
the same when the Temple was destroyed.14 Kristeva’s notion of the Temple enfolds onto our own bodies, in biblical
terms, the body as a Temple of the Holy Spirit: ‘A mouth attributed to the anus:
is that not the ensign of a body to be fought against, taken in by its insides.’15
This describes a possibility for blasphemy and sacrilege in how we occupy our
bodies individually, and by our relationship to other bodies. Seen from this
perspective, Piss Christ contravenes the boundaries of the self’s clean and proper
body, just as Ian Breakwell’s Deep Faith aligns throat and penis, as a symbolic
protest against an intrusive moral theology. Beyond the dichotomies of abjection and transcendence Kristeva’s project seeks to redeem the female body from categories of
abomination, from existing outside the Temple, due to functions that include
menstruation and lactation. And here it is helpful to imagine a fictitious artwork
by Serrano that elucidates Kristeva’s position, let us call it Menstrual Blood Christ. A crucifix submerged in menstrual blood indexes the sexualised female body, 154 Blasphemy and the art of the political and devotional identified as abject, and describes a problematic position for women in relation
to the holy place of the Temple. Serrano himself comes close to such a work with Milk, Blood, 1986. Two vats,
one of milk and the other of blood, are photographed butted up against each
other. Only the central vertical edges of the glass vats are visible so that the
image is abstracted and not dissimilar to some abstract colour field paintings. Serrano’s Milk, Blood, while engaging in binary opposites on a visual level,
encourages a more diffuse worry.16 For Anthony Julius, this work provides an
opportunity to reflect on the cultural connotations of bodily substances: ‘They
are similar: milk nurtures, blood animates. They are different: we ingest milk,
we celebrate its flow, we obtain it with a caress; we may not ingest blood (save
symbolically, or else to save our lives), we fear its flow, we obtain it painfully,
invasively...Like Piss Christ, the work is mediated by photography and presents
the same tensions between presentation and substance. We are seduced by the
formal, almost minimalist (Rothko) nature of the work but simultaneously
confronted by its substance’.17 Where Kristeva seeks to redeem the body from categories of abomination,
Carolyn Bynum Walker’s research on the female body and religious practices
in the later middle ages (1200–1500) promotes a bodily (incarnational)
manifestation of spiritual revelation that, I suggest, avoids binary opposition.18
She points to graphic physical processes being revered such as: ‘Holy people
spat or blew into the mouths of others to effect cures or convey grace. Beyond the dichotomies of abjection and transcendence The ill
clamoured for the bath water of would-be saints to drink or bathe in, and
preferred it if these would-be saints themselves washed seldom and therefore
left skin and lice floating in the water.’19 Following Francis of Assisi, who kissed
lepers, several Italian saints ate puss or lice from the poor or sick, thus
incorporating into themselves the illness and misfortune of others.20 Bynum
Walker cautions us against reading into medieval source material an assumption
of the binary that control of the body equates to a rejection of sex. She argues
that ‘Medieval images of the body have less to do with sexuality than with
fertility and decay. Control, discipline, even torture of the flesh is, in medieval
devotion, not so much the rejection of physicality as the elevation of it — a
horrible yet delicious elevation — into a means of access to the divine’.21 Two sixteenth-century religious paintings support this rejection of the body
as abject, and the connection between sexuality and abjection. Leo Steinberg
argues that, in Lucas Cranach’s Holy Trinity in a Glory of Angels (c.1515-18) and
Hans Schaufelein’s Crucifixion (1515) (both of which depict Jesus with an
erection), the erection is an iconographic symbol of the resurrection. A Dutch
painting, the Good Shepherd, c.1550, from the church of St John in Schiedam,
has Christ’s genitalia as the central focus of the pictorial space. St. Augustus
held that involuntary erections were a condition of original sin. Yet, in Christ’s 155 Negotiating the Sacred II body, as the New Adam, an erection is willed and intended, a sign of the
annulment of sin and death through the resurrection.22 To suggest a singularly
sexualised reading of such an image from the sixteenth century would be to
misread the material, and to fall into the binaries of much contemporary
discourse. Just as we do not need to read these binaries into the
crucifixion–erection–motif,
we
may
interpret
a
contemporary
Virgin–condom–motif as having to do with fertility and an elevation of
physicality as a means of access to the divine. Mieke Bal explores such a turning of the tables. According to Bal: The flesh that is so important in the Christian tradition takes on different
meanings at different moments in history. Beyond the dichotomies of abjection and transcendence From the perspective of an
engaging late twentieth century [and now the twenty first century]
where pain and suffering are often bound up with sexuality, we are now
enabled, by artists who endorse this baroque historiography and the
entanglement that characterises it, to scratch away the dust of a
disembodied religiosity and gain access again to a religious life that is
much closer to bodily experience.23 Cathy de Monchaux, Red, 1999, © Cathy de Monchaux Cathy de Monchaux, Red, 1999, © Cathy de Monchaux 156 Blasphemy and the art of the political and devotional In this light, another work from the Votive exhibition, Red, by Cathy de
Monchaux, challenges boundaries of the sacred and profane by adopting mixed
messages: heraldic motifs reminiscent of imperially religious crosses and crests,
and simultaneously, gynaecological-like instruments of fetish. Red collapses
institutional order into private blossomings of flesh. The sculpture is obsessively
created. With all the components cut by hand, the detailed clasps, buckles and
leather belts tie down, and yet follow the formation of a concentrically
descending vulva-like velvet interior. Like Ian Breakwell’s The Sermon, there is
a strongly public face to this work that nods to ecclesiastical and, in this case,
royal protocol. Desires of the flesh are here enfolded, literally and metaphorically,
onto the strictures of the law. But what is extraordinarily successful in this work
is the manner in which these conflicting elements never overpower each other
but are held together as palpable transgression. De Monchaux’s Red becomes a
powerful sign of the fertile body, the physically disciplined and restrained body
and, yes, the sexually engorged body, but the enriched body as signifier of
religious life in bodily experience. Bynum Walker’s caution against binary thinking is also helpful in discussing
Serrano’s work. If it were Christ’s piss (like the dirty bath water of would-be
saints), would the abject transfer to the realm of the sacred? What would we do
with a phial of Christ’s piss, other than venerate it? Moreover, does Serrano’s
photograph of Christ in piss, together with such uplifting and sublime aesthetics,
redeem piss from being an ensign of the body’s abject nature? Beyond the dichotomies of abjection and transcendence In this shift from
piss to a photograph of piss I have in mind Rosalind Krauss’ argument, in
referencing André Bazin’s ‘The Ontology of the Photographic Image’, that
photographs, by virtue of their process, are indexically linked to the objects
they portray in ways that exceed representation: ‘The photographic image is
the object itself.’ In this way, the photograph, together with its aesthetic qualities,
might redeem the abject substance.24 The act of pissing on the Koran in war
lacks the devotional and aestheticising intentions of both Serrano and Kovats. The meaning of the image Piss Christ is much more complex than the meaning
of either pissing on Christ or pissing on a crucifix, just as the meaning of the
sculpture Virgin in a Condom is more multifaceted than the significance of
comparing the Virgin Mary with an erection. Can we view these works from a
position that is beyond the conventional binary of the pure and impure? If we
consider the incarnational possibility that Christianity affords, to become as
Christ, then Serrano’s piss and Christ’s piss — Serrano’s body and Christ’s body
— may become as one. In the words of Damian Casey: ‘God’s place is with the
abject every bit as much as it is on the high altar of the cathedral.’25 In this
light, Piss Christ could be seen as ‘an attempt to retrieve the meaning of the
incarnation’.26 Casey contends that it is Serrano’s ‘exploration of the relation
between the abject and the sacred that makes Piss Christ not only good art, but
good religious art, bordering on the iconic’.27 157 Negotiating the Sacred II When Bynum Walker cautions a straight forward assumption of binary
thinking in medieval source material, she offers us a challenge to re-examine
works such as Piss Christ and Virgin in a Condom from perspectives that are not
preoccupied with the body as primarily sexual in function, or abject secretions. While both artworks are highly polemical and confrontational — this is piss
and this is a condom — both artists deny a singularly blasphemous intent,
alluding instead to both devotional and political intention. Both images have
historical precedents where body fluids and the body’s imagery are not clearly
divided between the sacred and the abject. I contend that such historical
perspectives inform a richer interpretation of Piss Christ and Virgin in a Condom. Beyond the dichotomies of abjection and transcendence Seen from this perspective, these images signify the possibility of divine
revelation through the body. Conclusion We cannot rely on a purely conventional understanding of blasphemy in
understanding blasphemy in artistic contexts. Intentions matter to our reading
of acts of blasphemy and change our perspective on whether they are acceptable. The context of the museum does more than reserve a space for artists to critique
outside the realms of worship, but enables the possibility of artists operating
across the parameters of critique and devotion. In this sense, the choice to curate
Votive: Sacred and ecstatic bodies in museums offered the possibility to interpret
Ian Breakwell’s Deep Faith and Cathy de Monchaux’s Red (and the other art
works discussed above) as ‘religious’ works that operate outside conformist
religious structures. In this respect museums and galleries have a role in
providing a space for the expression of alternative, and sometimes
unconventional, religious expression; offering artists, within certain restrictions,
the possibility to articulate what they want. The polemical nature of the abject
and sacred common to many of the artworks discussed in this chapter is not
new. Contemporary binaries regarding bodily fluids, suffering and flesh are
often bound up with sexuality, creating a sense of the religious that is
disembodied. It is in a reappraisal of our perceptions of binary imagery
(misunderstood by essentialist religious paradigms) that a solution emerges. Bynum Walker’s research into medieval perceptions of the abject body as less
sexual and more to do with fertility and decay indicate that the time at which
something is viewed changes our understanding of whether blasphemy has
occurred. It may be, that from the perspective of contemporary religious
authority, these images are blasphemous, however, historical readings of the
body in religious art make us question whether we must view works such as
Piss Christ and Virgin in a Condom in this fashion. These works operate within
a framework that is more complex than a conventional reading of blasphemy
allows. They may also be interpreted as devotional, religious expressions. 158 Blasphemy and the art of the political and devotional Blasphemy and the art of the political and devotional Endnotes
1 1 While the work of these artists provoked the greatest outcry, such juxtapositions of the sacred and
profane are also characteristic of the work of Ian Breakwell and Cathy de Monchaux discussed later in
this chapter in the context of the exhibition Votive: Sacred and ecstatic bodies. 2 The exhibition Votive: Sacred and ecstatic bodies toured two venues in New Zealand, the Adam Art
Gallery of Victoria University, Wellington, and the Dunedin Public Art Gallery, finishing in 2002. The
project was collaboratively curated by Mark Jackson as writer/curator and myself as artist/curator and
I acknowledge Mark Jackson’s assistance in thinking through many of the issues in this chapter as well
as Mel Hight, postgraduate candidate at AUT University. Votive responded to controversy surrounding
Tania Kovat’s exhibit Virgin in a Condom at the Museum of New Zealand Te Papa Tongarewa in 1998. Votive included works by: Ian Breakwell and Cathy de Monchaux (Britain); Pierre and Gilles (France);
Megan Jenkinson and myself (New Zealand). In choosing the works for Votive, we attempted to encourage
thinking beyond the binary of ‘right wing fundamentalism’ versus ‘freedom of speech’ and an
environment respectful of differences of opinion. 3 Kyla McFarlane, 2002, ‘Incisions and Excesses’, in Votive: Sacred and Ecstatic Bodies, Wellington and
Dunedin: Adam Art Gallery and Dunedin Public Art Gallery, pp. 10-19, at p. 10 4 McFarlane, ibid., note 2. 5 ‘Piss Christ exhibition 97.4’, The Arts Law Centre of Australia, 5 ‘Piss Christ exhibition 97.4’, The Arts Law Centre of Australia,
http://www.artslaw.com.au/reference/piss974/ (Viewed 27 April 2004.)
6 Ibid. http://www.artslaw.com.au/reference/piss974/ (Viewed 27 April 2004.)
6 Ibid. 7 McFarlane, op. cit., note 2. 8 8 ‘Anger over Virgin in a condom art’ 1998, Dispatch Online, 8 ‘Anger over Virgin in a condom art’ 1998, Dispatch Online,
http://www.dispatch.co.za/1998/03/11/foreign/condom.htm. (V 8 ‘Anger over Virgin in a condom art’ 1998, Dispatch Online,
http://www.dispatch.co.za/1998/03/11/foreign/condom.htm. (Viewed 20 December 2005.)
9 g
g
p
http://www.dispatch.co.za/1998/03/11/foreign/condom.htm. (Viewed 20 December 2005.)
9 9 Damien Casey, 2000, ‘Sacrifice, Piss Christ, and liberal excess’, p. 2,
http://dlibrary.acu.edu.au/staffhome/dacasey/Serrano.html. (Viewed 20 9 Damien Casey, 2000, ‘Sacrifice, Piss Christ, and liberal excess’, p. 2,
http://dlibrary.acu.edu.au/staffhome/dacasey/Serrano.html. (Viewed 20 December 2005.)
10 10 Stephen Davies, 1991, ‘Dickie’s Institutional Theory of the Definition of Art’, in Definitions of Art,
Ithaca, N.Y. Cornell University Press, pp. 78-114, at pp. 83-84. 10 Stephen Davies, 1991, ‘Dickie’s Institutional Theory of the Definition of Art’, in Definitions of A
Ithaca, N.Y. Cornell University Press, pp. Endnotes
1 78-114, at pp. 83-84. 11 McFarlane, op. cit., note 2, p. 13. 12 Curated by Bruce Fergusson and Sarah Olsen. 13 y
g
13 McFarlane, op. cit., note 2. 13 McFarlane, op. cit., note 2. 14 Julia Kristeva, 1982, Powers of Horror: An Essay on Abjection, Columbia: Columbia University Press,
p. 94. 15 Ibid
109 14 Julia Kristeva, 1982, Powers of Horror: An Essay on Abjection, Columbia: Columbia University Press,
p. 94. p
16 Anthony Julius, 2002, Transgressions: The Offences of Art, London: Thames and Hudson, p. 139. 17 Ibid. 18 Carolyn Bynum Walker, 1992, Fragmentation and Redemption: Essays on Gender and the Human Body
in Medieval Religion, New York: Zone Books, p. 162. 19 Ibid., p. 163. p
20 Ibid. 21 Ibid., p. 162. 22 Steinberg, Leo 1996, The Sexuality of Christ in Renaissance Art and in Modern Oblivion, Chicago: The
University of Chicago Press, pp. 318-324. University of Chicago Press, pp. 318-324. y
g
pp
23 Mieke Bal, 1999, Quoting Caravaggio: Contemporary Art, Preposterous History, Chicago: The University
f Chi
38
i d b
l
i
2 23 Mieke Bal, 1999, Quoting Caravaggio: Contemporary Art, Preposterous History, Chicago: The University
of Chicago Press, p. 38, cited by McFarlane, op. cit., note 2. Mieke Bal, 1999, Quoting Caravaggio: Contemporary Ar g
p
y
p
24 Rosalind Krauss, 1986, The Originality of the Avant-Garde and Other Modernist Myths, Cambridge,
Mass: MIT Press, p. 203. Mass: MIT Press, p. 203. 25 Casey, op. cit., p. 3. y
p
26 Ibid., p. 6. 26 Ibid., p. 6. p
27 Ibid., p. 2. p
27 Ibid., p. 2. 159 11
Negotiating the sacred body in Iranian
cinema(s): National, physical and
cinematic embodiment in Majid Majidi’s
Baran (2002) 11 Michelle Langford Any religious utterance, act or gesture, stands in the shadow of — more or
less, but never totally avoidable — perversion, parody and kitsch, of
blasphemy and idolatry.1 Any religious utterance, act or gesture, stands in the shadow of — more or
less, but never totally avoidable — perversion, parody and kitsch, of
blasphemy and idolatry.1 Where have all the bodies gone? Modesty in Iranian cinema In the post-revolutionary period, Iranian cinema has been subject to strict Islamic
censorship regulations that dictate what can and cannot be show on screen,
particularly in terms of the representation of women and romantic relationships
between men and women. The rules relating to the representation of women
mirror, in a highly exaggerated way, the codes of modesty relating to women
in society more generally. Censorship functions to ensure that both onscreen
bodies and the bodies of viewers in the audience remain modest. These bodies
must be protected from un-Islamic, profane, blasphemous or sacrilegious
utterances, acts and gestures: primarily those related to sex and un-Islamic
behaviour. This is achieved in life primarily through the veiling of female bodies,
and on screen by a ‘screening’ of such acts and suggestions of sexuality that
might cause one to become impure or ‘haram’. The codes of modesty that prevail
in society are greatly enhanced in the cinema, as all viewers are considered to
be unrelated to the actors and characters who appear on screen. Modesty is
maintained primarily by ensuring that onscreen women are appropriately veiled
and clothed in loose-fitting clothing, and that physical contact between male
and female bodies is avoided. Stories involving either emotional love or the
suggestion of physical love between members of the opposite sex are generally
avoided primarily due to the risk of violating the sanctity of pure, modest and
Islamic bodies.2 Film form, that is, the technical and stylistic devices for creating
meaning in cinema, according to Hamid Naficy has also undergone a radical
process of Islamicisation, which obliges filmmakers to uphold the principles of
Islam.3 In a religious context, where idolatry and imitating the creative act of 161 Negotiating the Sacred II God is considered to be highly sacrilegious, it is a wonder that the creative
audio-visual medium of film has been adopted so readily as an instrument for
consolidating the revolution and educating the population about proper Islamic
morals and behaviour. Where have all the bodies gone? Modesty in Iranian cinema Indeed, despite his constant critique of cinema during
his years in exile, Ayatollah Khomeini is said to have seen the potential of the
film medium for promoting revolutionary (primarily Islamic) values.4 Just as
Western cinema and the popular and decadent pre-revolutionary cinema known
as ‘Film Farsi’ were seen as an evil force that had corrupted the minds and bodies
of the population during the Phalavi regime, in the post-revolutionary period,
cinema was considered an appropriate vehicle for the purification of spectator’s
and by association the nation’s mind, body and soul. With the enmeshing of
state and religion brought about with the establishment of the theocratic
government in 1979 and the establishment of Shari’a law, both individual and
nation must necessarily reflect Islamic values. Majidi Majidi’s Baran (2001) is an extraordinary film which successfully
depicts an adolescent man discovering his burgeoning sexuality, and falling in
love with a beautiful young Afghan woman named Baran. Through the use of
very clever and subtle cinematic devices, Majidi generates a deep sense of
emotion and intimacy without ever showing any physical contact between the
two, and without ever violating either the character’s or the viewer’s modesty. Additionally, by couching this love story in the broader context of the
socio-economic condition of Afghan refugees in Iran, Majidi produces yet another
level of discursive meaning where an ideal model of Islamic love and charity
toward others may be perceived, effectively embodying a highly idealised
conception of the nation. But, through its complex visceral engagement of the
spectator’s sensorium, Baran is a film that also explores the very limits of the
sacred Iranian body on both the level of the national and individual citizenry. In this chapter, I wish to argue that through the simultaneous idealised
embodiment of Iranian national Islamic values and the faltering, frail and very
physical adolescent body of the central male character, Majidi is effectively
negotiating the limits of the sacred body in Iranian cinema(s). I wish finally to
demonstrate how film — which itself may be considered a body of sorts — may
reach out figuratively to touch and move the bodies in the audience in ways
that exceed simple observance of censorship regulations and generate vital
connections between the viewers and the bodies on screen. In doing so, the film
bears witness to the shadows that haunt any sacred utterance. Embodying the nation We see just such a reflection of individual and nation in the narrative trajectory
of Baran. On one level, Majidi’s film functions as an exemplary model of selfless
devotion and modesty, particularly through its characterisation of the central
male character Lateef, an adolescent gofer working on a building site on the 162 Negotiating the sacred body in Iranian cinema(s) outskirts of Tehran. The film traces his growing attachment to a young Afghan
woman named Baran, who has come to the building site disguised as a young
man named Rahmat in order to support her family following an accident that
has rendered her father physically unable to work. Once Lateef discovers that
Baran is really a woman, he is clearly struck by love’s arrow and goes out of his
way to please her and swoons in her presence. Eventually, however, Baran is
forced to flee the building site as an illegal Afghan worker and Lateef goes in
search of her. When he discovers the poor conditions in which she lives, and
the harsh working conditions she endures, hauling large rocks from a rushing
stream, he resolves to help her and her family by giving them all of his savings. Things do not go to plan, however, for Baran’s father, Najaf, gives the money
to his friend, Soltan, who needs it to return urgently to Afghanistan. Surprisingly,
Lateef is not embittered by this situation, for he recognises that this man is just
as desperate as Baran and her family. This is the first example of the
self-sacrificing behaviour exhibited by Lateef throughout the film, putting
collective needs above his own personal desires. Lateef’s behaviour also functions powerfully as an illustration of the Islamic
principle of welcoming Muslim refugees and displaced peoples. During the
Soviet occupation of Afghanistan (1979–89), Iran opened its borders to Afghan
refugees who were classified by the newly formed Iranian government as
Mohajerin, or ‘involuntary religious migrants’.5
This principle, which is
enshrined in the Quran was, according to Aldeeb Abu-Sahlieh, adopted in the
second Islamic Declaration of Human Rights in 1981: The homeland of Islam (dar-al-Islam) is one. It is a homeland for every
Muslim, whose movement within [its domain] cannot be restricted by
any geographical impediments nor political boundaries. Embodying the nation Every Muslim
country must receive any Muslims who emigrate thereto, or who enter
it, as a brother welcomes his brother: ‘Those who entered the city and
the faith before them love those who flee unto them for refuge, and find
it in their breasts no need for that which had been given them, but prefer
the fugitives above themselves though poverty become their lot. And who
is saved from his own avarice – such are they who are successful’... [Quran, 59:9]6 Lateef’s first act of self-sacrifice, of preferring the fugitives above himself
despite the certainty that this will increase his own poverty, tends to cast him
as an ideal figure of the Islamic nation as the Quranic principles are powerfully
embodied in him. This embodiment of Islamic values is further emphasised in
his second self-sacrificing act, which becomes for Lateef an act of effacing his
own individual identity. Throughout the film, Majidi places emphasis on the themes of identity and
identification in a number of ways that provide commentary on Iranian civil 163 Negotiating the Sacred II society and the shifting policies toward Afghan refugees in that society. The
first instance occurs during the first few minutes of the film in a scene where
Lateef goes to a supermarket to purchase food for the construction workers. Lateef has had to leave his identity card in order to secure credit at the store. The shopkeeper points out that the account is long overdue and demands
payment. At risk is Lateef’s identity card, and we recognise that he risks the
security of his personal identity for the good of the collective (the labourers). Later, as Lateef is checking into a hotel, we see a man denied a bed for the night
because he does not have an identification card. Additionally, on two occasions,
the illegal Afghan workers (who do not have work permits) are forced to flee
the building site when a government inspector comes to check for illegal workers. Embodying the nation It becomes clear, when the foreman of the site is asked to sign a document stating
that no illegal Afghan workers are employed on the site, that these Afghans are
no longer treated as mohajerin who ‘were issued with identification cards known
as “blue cards”, and granted indefinite permission to stay in Iran legally’.7 This
detail of the film closely reflects Iran’s changing policies toward Afghan refugees,
who, after 1993, were no longer categorised as mohajerin, but simply as
panahandegan (refugees), a term which ‘was considered to have a pejorative
nuance, even connoting impoverishment’.8 These panahandegan were issued
only with temporary registration cards, which made access to work and other
civil services such as health care and education extremely limited. Lateef’s strong
sense of ideal Islamic devotion to the ‘fugitives’ is clearly being contrasted here
with a view of the state’s pragmatic approach to the refugee question.9 By
extension, this would appear to suggest that, while the government of the Islamic
Republic of Iran is constitutionally bound to abide by the principles of Islamic
law, this may not necessarily be the case in practice, given the explicit removal
of sacred significance from the Afghan refugees, which seems to ignore the
passage from the Quran cited above. Majidi certainly provides an ideal role
model in his film, however, by revealing a considerable gap between this ideal
behaviour and the practices of the state, he has presented us with an allegorical
figure aimed at representing the opposite of that which he would appear to
represent. Lateef’s behaviour is revealed to be explicitly and unrealistically
ideal. The presentation of the ideal individual is used to highlight the flaws of
the state. Throughout the film, Lateef continues to practice such sacred beliefs toward
his fugitive neighbours. This is brought to a climax with a highly symbolic and
self-effacing gesture. Having already given away his entire savings, Lateef
decides to sell his identity card in order to provide Baran and her family with
the money they need to return to Afghanistan. In aesthetic terms, Majidi sets
up this scene as though Lateef is making a dangerous journey into the
underworld, further emphasising the austere and self-sacrificial nature of his
gesture. As Lateef makes his way through the busy bazaar making enquiries, 164 Negotiating the sacred body in Iranian cinema(s) suddenly, two mysterious men begin to chase him through the narrow alleyways
of the bazaar. Embodying the nation But, after successfully giving them the slip, another man approaches
him and urges Lateef to follow. He leads Lateef to a dark, cluttered old basement,
which further emphasises the theme of his descent into the corrupt underbelly
of Tehran. Here, another man purchases his identification card. A close-up of
the card signals the symbolic intensity of this moment, as the man removes
Lateef’s photo, and metaphorically erases his identity with a quick flick of his
grubby finger. This act of selling his identity card is a powerful gesture in which
Lateef shows his preference for the fugitives above himself, even though this
will certainly ensure his own poverty. Lateef is the very embodiment of the
ideal national subject envisaged for the post-revolutionary Iranian cinema by
Khomeini, yet ironically his perfection reveals the cracks and contradictions,
the shadow of the very nation-state he is supposed to embody. Lateef’s developing model behaviour effectively serves as a critique of, rather
than as an example of, state practice. Furthermore, by choosing to relinquish
his identification/identity he is effectively rendered a non-citizen, like the Afghan
refugees, and therefore it is questionable whether he even stands for the state
after all, but rather becomes one of the many displaced people of the region. Adolescent awakenings Even before the introduction of Baran/Rahmat to the film, we are provided with
a glimpse of Lateef’s burgeoning but floundering interest in the opposite sex. On his way back to the building site after shopping for bread, his attention is
drawn by the sound of a woman giggling suggestively. We see him looking
wistfully for a few seconds before the object of his gaze is revealed to be a young
couple in a park playfully throwing a hat to one another. Such a playful but
mediated tryst serves frequently in Iranian cinema as a coded substitute for a
more intimate exchange, which is forbidden from the screen. The
shot-reverse-shot structure of this brief scene leaves the viewer with no doubt
that Lateef also longs for such female companionship as he chews distractedly
on a piece of Afghan bread. Where the hat serves as a playful indirect signifier
of forbidden physical contact — a reminder that the couple are not permitted
to hold hands in public — the fleshy bread may be read as a cryptic clue or
allegorical hieroglyph of Lateef’s burgeoning desire to connect with another
body. In fact, the bread is charged with such allegorical meaning from the very
beginning of the film, when the viewer becomes attuned to the inherently fleshy
qualities of raw dough. This is initially achieved through the evocative
introduction of the soundtrack even before the first image has lit the screen. For
a few seconds we hear some slapping sounds, not unlike the sound of a hand
gently rubbing and slapping flesh. This is accompanied by breathing sounds,
which are pushed to the very front of the soundscape, giving the cinema viewer
the rather tactile sensation that someone is breathing in our ear. As with the
momentary visual withholding of the source of the giggling woman, the few
seconds during which vision is withheld allows the viewer’s mind to wander,
to invest these sounds with our own imaginations, to speculate on the kinds of
images that may accompany them. In this sense, the black screen functions as a
kind of veil,11 highlighting that something has been hidden from view, and
signalling that we must engage both our minds and senses in order to delve into
the deepest layers of meaning presented by the film. From embodied nation to embodied desire If Majidi presents Lateef as embodying the ideal, self-effacing values of Islam
through his devotion to a series of Afghan refugees on a socio-political level,
and, as I have shown it is possible to read this as signifying the opposite of what
it appears to, then we need to investigate the various levels at which embodied
desire is simultaneously effaced and achieved through the use of cinematic
devices and a very complex deployment of the sound track. Despite the necessary effacement of the physical, sexualised body in Iranian
cinema according to the mandates of censorship, throughout Baran, Majidi
manages powerfully to suggest the sexual awakening of a young adolescent man,
taking him through stages of confused outbursts of anger in the all-male
environment of the building site, to the discovery of his own capacity for love
of a woman. By staging this inner journey on a building site — ostensibly a male
dominated arena — in which a woman (Baran/Rahmat) intervenes, Majidi is
effectively attempting to argue for the necessity of interaction between men and
women in order for appropriate subject formation to take place and respect for
one another to be achieved. Ironically, however, the woman arrives disguised
as a man. Certainly, throughout the film Baran is literally figured as a
domesticating, ordering and beautifying influence upon this rather ramshackle
and chaotic space. Michael Fischer has suggested that Baran functions
metaphorically as a cooling influence on Lateef’s hot-blooded adolescent
temper,10 although, I would argue, not without first firing him up. Thus, the 165 Negotiating the Sacred II film skirts the very edges of the sacred by alerting us to the pleasures of the
flesh. Adolescent awakenings Following this aural ‘tease’,
Majidi provides an answer to the sound puzzle, revealing the source of the
sounds to be a baker kneading dough, rolling it out and slapping it onto the hot
stone to be cooked. Even though this image manages to dispel the more titillating,
and potentially sacrilegious suggestions provided by the sounds, Majidi has
managed to attune the viewer to the particularly fleshy qualities of the dough,
and predisposed us to read sensuality into Lateef’s consumption of it in the scene
described above.12
L t
f’
k
i
d l
t d
i
i f
th
h
i
d
t l t Lateef’s awakening adolescent desire is further emphasised moments later
when he pauses to admire himself in a highly reflective glass door, fixing his 166 Negotiating the sacred body in Iranian cinema(s) hair, smiling and humming to himself, imagining perhaps that a girl might find
him attractive. However, this moment of what may be described as narcissistic
scopophilia — where he already perceives himself to be more man than he really
is — is interrupted (metaphorically shattered) when a man pushes open the door
from the inside, pauses to look at him suspiciously, thus bringing Lateef’s
self-reverie to an abrupt halt. His self-image is not yet fully formed as he balances
on the cusp between adolescence and adulthood. These are just the first of many clues that hint at Lateef’s sexual awakening. On the building site, we witness other signs of his faltering and often misplaced
attempts to assert his burgeoning masculinity. This fire that is burning inside
him seems to function at times — such as those discussed above — as a calm,
warming sensation, but at other times will flare out of control and provoke
violent outbursts. Lateef frequently fights with his co-workers over highly
insignificant matters, particularly once the young Rahmat (Baran disguised as a
young man) comes to work on the building site. These outbursts become more
frequent, particularly after Rahmat is given his job in the kitchen preparing
meals and tea and Lateef must join the ranks of the labourers. Gradually, his
outbursts become more focussed on Rahmat, showing ironically that he is learning
to direct his adolescent rage. This rage, however, is suddenly turned to desire
in a key moment when Lateef discovers that Rahmat is really a woman. Adolescent awakenings This scene is important for the way it is structured cinematically, for it is
designed to generate a powerfully affective experience for the spectator as well
as revealing the emotions that are being stirred up deep within Lateef’s being. It also cleverly negotiates the censorship rules regarding the portrayal of women
in Iranian cinema. The scene opens with a shot of Lateef going to fetch a heavy
bag of cement. As he moves toward the camera, struggling to carry the heavy
bag in his arms, wind blows a wave of white smoke across the screen, which
irritates Lateef’s eyes, causing a momentary loss of vision. The sound of the wind
generates a highly mystical effect, appearing as a kind of energy that draws
Lateef toward the kitchen area where Rahmat/Baran works. The soundtrack is
then further layered with the distant sound of thunder, rain, and the faint sound
of a woman humming. As Lateef blinks, a reverse shot of a curtain blowing in
the wind briefly reveals the distant silhouette of a woman. Cut back to Lateef,
still blinking he throws the bag of cement to the ground and rubs his eyes. The
sound of wind continues to intensify, and having regained his vision, Lateef
peers intensely toward the blowing curtain, the camera zooming toward him to
indicate the sharpening of his vision. This movement is answered in the next
reverse shot of the curtain, which now blows in slow motion, teasingly rising
and lowering, providing another brief shot of the silhouette glimpsed earlier,
but the curtain refuses to yield for more than a second or two. The movement
and placement of the camera suggests a point of view shot, however when Lateef
enters from screen right, we realise that Majidi has in fact placed us not with 167 Negotiating the Sacred II Lateef, but beside him, therefore implicating us within the scene. The following
shot, however, does not yet reveal to us a clearer view of the scene behind the
curtain, we see Lateef seeing, his eyes widening, fixing his gaze upon the scene
before him. Finally, Majidi provides us with a shot of Baran brushing her long
black tresses, humming to herself wistfully as she does so. Adolescent awakenings She is framed and
‘veiled’ by a frosted window that barely separates either Lateef or the viewer
from her modesty with backlighting providing a further distancing effect to
protect the modesty of all. In this highly affective scene produced by sound,
framing, camera movement and movement within the frame, not only are we
encouraged to feel the passion that Lateef feels, but to experience images and
sounds viscerally. Don’t tell the mullahs! According to Sobchack, coenaesthesia is a common, yet under-recognised
perceptual experience in which ‘our equally available senses have the capacity
to become variously heightened and diminished’.18 This capacity is more evident
in children where the hierarchical socialisation of the senses has not yet fully
taken place, but given the right stimuli, this may also occur in adults. Cinema
has the most wonderful capacity to do this, and certainly, as I have already
shown, through his complex use of sound19 in Baran, Majidi certainly attempts
to retrain his viewers’ sensorium to privilege sound over or in addition to vision. This adds a highly affective and potentially erotic or subversive level of meaning
to the film that plays off and against the pure, selfless devotion on a collective
level, and the personal love for a woman on an individual level. It is my contention that a figurative and literal exchange between potentially
erotic bodies takes place in Baran. As in the scene discussed above, Majidi
deploys complex cinematic techniques (sound, editing, camera and character
placement) and stimulates our sensory organs (through evocative sounds and
images) in order to weave the viewer into the very texture of the film. In doing
so, he effectively allows the viewer to experience that which cannot be literally
represented on screen. Our bodies effectively fill the gap imposed by censorship
between the characters’ bodies. In doing so, Majidi exposes the viewer to the
possibility of what I would like to call ‘sacrilegious affects’, that is, a felt violation
of the sacred modesty of the Iranian cinema screen. The cinema’s potential for
cross-modal transfer enables the superficial modesty of the on screen
representation to be undermined by a transgressive potential in the aesthetic
realm, capable of activating the forbidden (haram) sensations of the flesh. Throughout the film, the sounds of wind, rain, thunder, running water, the
fluttering of fabric and bird’s wings, human breath, footsteps, voices, laughter
and birds singing are all used to heighten the embodied sensory perceptions of
the viewers and attune them to what must remain unrepresented, relegated to
the space beyond the frame. Even a close-up of Lateef’s finger wiping mud from
a coin he finds in the street works to heighten our sense of touch. Don’t tell the mullahs! Throughout Baran, Majidi effectively uses images and sounds to engage the
viewer in what Vivian Sobchack refers to as a ‘cinesthetic’ mode of embodied
spectatorship.13 In her article, ‘What my fingers knew’, Sobchack argues that
to experience a film is not simply a matter of ‘seeing it’. Rather, our body
functions as ‘a ‘third’ term that both exceeds and yet is within representation’.14
That is, although our bodies are ostensibly located outside and separate from
the representations that appear on screen, through the cinema’s engagement of
our multiple senses, our bodies are necessarily also inscribed into or take part
in the representation itself. In addition, she writes: ‘All the bodies in the film
experience — those on-screen and off-screen (and possibly that of the screen
itself) — are potentially subversive bodies. They have the capacity to function
both figuratively and literally.’15 In coining the term ‘cinesthesia’, Sobchack
draws on two psychoneurological conditions: synaesthesia and coenaesthesia. Synesthesia is an extreme, but rare condition in which the stimulation of one
sense provokes a perception in another sense. Sobchack explains: ‘Synaesthetes
regularly, vividly, automatically, and consciously perceive sound as colour, or
shapes as having a taste’.16 A less extreme form of this ‘cross-modal transfer’
takes place in figural language. According to George Lakoff and Mark Johnson,
because metaphors originate in concrete, sensate experience, ‘metaphor is
experiential and visceral’.17 I would argue that the highly metaphorical film
language used in Baran, combined with the clever use of off-screen sound, helps
to engage the viewer in just this kind of cross-modal transfer. Not only does
bread metaphorically evoke flesh, it does so largely because we have already
heard/felt the potentially erotic fleshiness of the dough well before our sense of
sight is brought into play. Similarly, in the scene in which Lateef discovers
Baran’s true identity, the enhanced sound of wind serves simultaneously as a
metaphor for the swell of emotion being experienced by Lateef, and may cause
a cross-modal sensation in the viewer of being touched by such a breeze. 168 Negotiating the sacred body in Iranian cinema(s) The dominant role played by sound in producing these cross-modal sensory
affects in Baran also evokes the second bodily condition discussed by Sobchack. Don’t tell the mullahs! By the end of
the film, our senses and emotions have become so heightened that we are
prepared for the emotionally (and sexually) charged scene that closes the film. In this scene, Baran and her family are preparing to leave for Afghanistan and
Lateef has come to their village to help. As Baran crosses the grey, muddy
pathway she trips and the contents of her wicker bag spill out onto the ground. This functions as a moment of metaphorical explosion, betraying perhaps the
feelings she has kept hidden throughout the film. We see among a variety of
other fruits and vegetables, several bright red tomatoes, and some dried figs. Lateef rushes to Baran’s aid and Majidi cuts to a close-up filmed in slow motion
of Baran and Lateef picking up the spilled items. As their hands slowly and 169 Negotiating the Sacred II gracefully enter the frame a moment of tactile pleasure may be experienced by
the viewer. Although, according to the rules of modesty, the characters do not
physically touch, the framing of the image enables their images to overlap as
they each reach for a ripe red tomato. They therefore ‘touch’ virtually in this
brief and silent parting exchange, which is heightened by the contrast between
the grey earth and the redness of the tomatoes, which provide a melodramatic
sign of their passion. Here, the modest, restrained emotions depicted on screen
overflow into the space between the viewer and the screen, into the dark space
of the cinema where we may even ‘steal’ a publicly forbidden tactile moment. The almost-direct looks of the protagonists toward the camera that directly
following this scene invite us to share in this intimate moment, further charging
this affective relationship between the screen and the viewer. While, as I have
shown, Baran adheres very strictly to Islamic censorship regulations, this
relationship between screen and spectator set up by the film teeters on the
precipice between the sacred and the sacrilegious, effectively pushing the limits
of Islamic censorship that prevents intimacy between men and women from
being depicted on screen. The modest bodies on screen are shadowed by the
bodies of the spectators seated in the dark space of the cinema. But hush! Please
don’t tell the mullahs, because Iranian filmmakers have enough trouble with
censorship as it is! 1 Hent de Vries, 1999, Philosophy and the Turn to Religion, Baltimore: Johns Hopkins University Press,
p. 11. 2 More recently, however, numerous films have dealt with the subject of romantic love, including the
present film under discussion. Some of these films, which also fall into the genre of the ‘spiritual film’,
use romantic love as a means for discovering divine love. Many take traditional Persian Sufi poetry as
their model, pivoting around an ambiguous presentation of earthly and divine love. A recent example
of such a film is God is Near (Khoda Nazdik Ast, Ali Vazirian, 2007). Endnotes
1 1 Hent de Vries, 1999, Philosophy and the Turn to Religion, Baltimore: Johns Hopkins University Press,
p. 11. 1 Hent de Vries, 1999, Philosophy and the Turn to Religion, Baltimore: Johns Hopkins University Press,
p. 11. 2 More recently, however, numerous films have dealt with the subject of romantic love, including the
present film under discussion. Some of these films, which also fall into the genre of the ‘spiritual film’,
use romantic love as a means for discovering divine love. Many take traditional Persian Sufi poetry as
their model, pivoting around an ambiguous presentation of earthly and divine love. A recent example
of such a film is God is Near (Khoda Nazdik Ast, Ali Vazirian, 2007). 3 Hamid Naficy, 2002, ‘Islamicizing Film Culture in Iran: A Post-Khatami Update’ in Richard Tapper
(ed.) The New Iranian Cinema: Politics, Representation and Identity, London & New York: I.B. Tauris,
pp. 26-65. 4 3 Hamid Naficy, 2002, ‘Islamicizing Film Culture in Iran: A Post-Khatami Update’ in Richard Tapper
(ed.) The New Iranian Cinema: Politics, Representation and Identity, London & New York: I.B. Tauris,
pp. 26-65. 4 3 Hamid Naficy, 2002, ‘Islamicizing Film Culture in Iran: A Post-Khatami Update’ in Richard Tapper
(ed.) The New Iranian Cinema: Politics, Representation and Identity, London & New York: I.B. Tauris,
pp. 26-65. 4 4 This point is mentioned by numerous commentators including Richard Tapper in his introduction to
The New Iranian Cinema, pp. 5-6; and Hamid Dabashi, 2001, Close Up Iranian Cinema Past, Present and
Future, London: Verso, p. 32. 4 This point is mentioned by numerous commentators including Richard Tapper in his introduction to
The New Iranian Cinema, pp. 5-6; and Hamid Dabashi, 2001, Close Up Iranian Cinema Past, Present and
Future, London: Verso, p. 32. 5 Mohammad Jalal Abbasi-Shavazi, Diana Glazebrook et al., 2005, ‘Return to Afghanistan? A Study of
Afghans Living in Tehran’ Afghanistan Research Evaluation Unit, Faculty of Social Sciences, University
of Tehran, p. 12. 5 Mohammad Jalal Abbasi-Shavazi, Diana Glazebrook et al., 2005, ‘Return to Afghanistan? A Study of
Afghans Living in Tehran’ Afghanistan Research Evaluation Unit, Faculty of Social Sciences, University
of Tehran, p. 12. 6 Sami A. Aldeeb Abu-Sahlieh, 1996, ‘The Islamic Conception of Migration’ The International Migration
Review, vol. 30, no. 1, pp. 37-57. p. 53. Emphasis added. 6 Sami A. Aldeeb Abu-Sahlieh, 1996, ‘The Islamic Conception of Migration’ The International Migration
Review, vol. 30, no. 1, pp. 37-57. p. 10 Michael M. J. Fisher, 2004, Mute Dreams, Blind Owls, and Dispersed Knowledges: Persian Poesis in
the Transnational Circuitry, Durham & London: Duke University Press. The metaphorical significance
of Baran’s name is twofold. It is at once the name of the river running through Karbul in Afghanistan
and means ‘rain’ in Persian. 8 Ibid., p. 13. 9 It should also be noted that the manager of the building site also treats the Afghans as mohajerin,
paying them before the Iranian workers, much to the chagrin of those workers. 7 Abbasi-Shavazi et. al., op. cit., p. 12. 18 Sobchack, op. cit. 19 There are numerous other examples in the film, which I do not have space to discuss here. Endnotes
1 53. Emphasis added. 7 Abbasi-Shavazi et. al., op. cit., p. 12. p
9 It should also be noted that the manager of the building site also treats the Afghans as mohajerin,
paying them before the Iranian workers, much to the chagrin of those workers. p
9 It should also be noted that the manager of the building site also treats the Afghans as mohajerin,
paying them before the Iranian workers, much to the chagrin of those workers. 10 Michael M. J. Fisher, 2004, Mute Dreams, Blind Owls, and Dispersed Knowledges: Persian Poesis in
the Transnational Circuitry, Durham & London: Duke University Press. The metaphorical significance
of Baran’s name is twofold. It is at once the name of the river running through Karbul in Afghanistan
and means ‘rain’ in Persian. 170 Negotiating the sacred body in Iranian cinema(s) Negotiating the sacred body in Iranian cinema(s) 11 Indeed, the blackness of the screen reminds me of the chador, the Islamic veil which is commonly
used by devout Muslim women in Iran, and are required to be worn in all sacred spaces such as mosques
and courtrooms. The chador is predominantly black, although a range of floral variants are often seen. 12 Later in the film, Majidi employs a process of metaphorical slippage, which functions to confirm the
allegorical (rather than statically symbolic) nature of the aural and visual imagery. The complex
soundscape of the building site recalls the opening sounds, and a number of times we see Baran and
other characters mixing cement by hand, recalling the tactile process of making bread by mixing flour
and water. Thus both bread and cement may substitute for forbidden physical contact between men 11 Indeed, the blackness of the screen reminds me of the chador, the Islamic veil which is commonly
used by devout Muslim women in Iran, and are required to be worn in all sacred spaces such as mosques
and courtrooms. The chador is predominantly black, although a range of floral variants are often seen. 12 Later in the film, Majidi employs a process of metaphorical slippage, which functions to confirm the
allegorical (rather than statically symbolic) nature of the aural and visual imagery. The complex
soundscape of the building site recalls the opening sounds, and a number of times we see Baran and
other characters mixing cement by hand, recalling the tactile process of making bread by mixing flour
and water. 17 Lakoff and Johnson quoted in Sobchack, ibid. Endnotes
1 Thus both bread and cement may substitute for forbidden physical contact between men
and women. 13 Vivian Sobchack, 2000, ‘What My Fingers Knew: The Cinesthetic Subject, or Vision in the Flesh’
Senses of Cinema, vol. 5. http://www.sensesofcinema.com/contents/00/5/fingers.html. (Viewed 6 July
2004.)
14 Ibid. 15 Ibid. 16 Ibid. 17 Lakoff and Johnson quoted in Sobchack, ibid. 18 Sobchack, op. cit. 19 There are numerous other examples in the film, which I do not have space to discuss here. 11 Indeed, the blackness of the screen reminds me of the chador, the Islamic veil which is commonly
used by devout Muslim women in Iran, and are required to be worn in all sacred spaces such as mosques
and courtrooms. The chador is predominantly black, although a range of floral variants are often seen. 12 Later in the film, Majidi employs a process of metaphorical slippage, which functions to confirm the
allegorical (rather than statically symbolic) nature of the aural and visual imagery. The complex
soundscape of the building site recalls the opening sounds and a number of times we see Baran and 11 Indeed, the blackness of the screen reminds me of the chador, the Islamic veil which is commonly
used by devout Muslim women in Iran, and are required to be worn in all sacred spaces such as mosques
and courtrooms. The chador is predominantly black, although a range of floral variants are often seen. 12 Later in the film, Majidi employs a process of metaphorical slippage, which functions to confirm the 12 Later in the film, Majidi employs a process of metaphorical slippage, which functions to confirm the
allegorical (rather than statically symbolic) nature of the aural and visual imagery. The complex
soundscape of the building site recalls the opening sounds, and a number of times we see Baran and
other characters mixing cement by hand, recalling the tactile process of making bread by mixing flour
and water. Thus both bread and cement may substitute for forbidden physical contact between men
and women. 13 Vivian Sobchack, 2000, ‘What My Fingers Knew: The Cinesthetic Subject, or Vision in the Flesh’
Senses of Cinema, vol. 5. http://www.sensesofcinema.com/contents/00/5/fingers.html. (Viewed 6 July
2004.) 19 There are numerous other examples in the film, which I do not have space to discuss here. 171 171 12
Silence as a way of knowing in Yolngu
Indigenous Australian storytelling Caroline Josephs I am, in this chapter, approaching three aspects of silence as a way of knowing
— in relation to Yolngu story and storytelling — through one particular Yolngu
story which I cannot tell. What I can tell — is why I cannot tell the story, and
how pursuing the question of whether I could tell the story, and in what way,
led me on a long intriguing journey. The three aspects I want to deal with, in relation to one Yolngu story, are: 1. protocols of being silent around storytelling — or not telling; 2. inside/outside knowledge, and touching into the silence surrounding,
and in women’s business; and 3. embodiment as a silent way of knowing when dancing country. 3. embodiment as a silent way of knowing when dancing country. The way of storytelling My approach is a step towards Yolngu ways of being and telling as far as may
be possible for a non-Indigenous non-Yolngu storyteller and researcher, and to
make the bridging necessary for a similar audience as myself. Yolngu
epistemology is part of what I am terming sacred epistemology — a mystical
tradition which includes phenomena such as revelation and epiphany, and of
being present to the seen, the known, and at the same time, to Mystery, the
Unknown, the ‘between’, the ‘ship’ of relating. In this process, being becomes
Being, for Yolngu. It is also primarily an experiential approach to knowing, a
felt sense in the body, and can not in my view, be met through analytic
conceptual frames of reference. According to Greg Dening,1 story is the only
way such a sacred world may be represented. Current experiences are sung and
told as contemporary expressions of the Dreaming, the moving from the Wangga
to the present. Past and present thus become fused and ensure a future of deep
intimacy with all beings. My way of presentation is to work towards exemplifying such approaches
— through personal story (never separate from Yolngu ‘history’ or Dreamtime
story) and traditional storytelling, in ways that represent relationship with
country, and with all beings, including kin with others of clan and tribe. I weave 173 Negotiating the Sacred II these storytellings with explanation in the non-Yolngu tradition to make the
connections clearer for we who are non-Indigenous readers. Knowing, for Yolngu,
is always being approached as an interior experience for the individual, albeit in
the context of the tribe’s cultural knowledge — it is as a consequence always
done within silence. Outside or exterior experiences will always be parallelling
interior internal experiences in symbolic ways, in feeling states and in states of
mind. Storytelling is always intending to link each Yolngu to their own country,
to plants, to animals, to all the Dreamings, and to each other, in particular kinship
interrelationships which are articulated not only through storytelling, but are
constantly reinforced through an elaborate and refined ‘opera’ of mark-making,
music-making, protocols of relating presented in everyday life and ceremony
both. (If you are my mother-in-law for example, there are certain kinds of body
language to be presented in your company, and only certain kinds of humour
may be displayed with certain family members). The way of storytelling So, I am presenting a
methodology pointing towards Yolngu ways of knowing at the same time as
telling the stories. The people, the place, the story Yolngu people are Indigenous Australians living on their land which is in
north-east Arnhemland, of Australia. Australian Aboriginal people have been
on their land continuously for perhaps 40,000 or more years. (Yolngu language
is probably around 5000 years old). Yolngu came into first contact with white
people in the 1920s with a few anthropologists who travelled by boat to the
remote area, in the 1930s with the establishment of a mission school, and with
the impact of the multi-national bauxite miners, in the 1960s. As a necessary precursor I am telling of the journey that led me to this Yolngu
story. The story, commonly called in Arnhemland, The Wagilag Sisters Story,
is a story of the Rainbow Serpent. Yolngu personal stories are bound together
with ancestral stories. Narrative (or ‘story’) is inextricably woven with country,
and body to country, and all beings to each other and to country through the
laws of kinship. In 1979 I met Wandjuk Marika, an Elder, important Law man, master yidaki2
player, artist, from north-east Arnhemland — at a conference in Darwin where
I was talking of the project I was running at the time across Australia. I was
offering a presentation on ‘The Curriculum Development Centre’s Multi-Arts
Project’ in terms of the rainbow, with its many colours and strands. After,
Wandjuk came up to me, shook my hand, and thanked me for the story of the
Rainbow. We sat in the garden for several hours (I don’t recall how long — clock time
ceased) as he told me stories of his life and, in the course of this, gave my children
Aboriginal names. (It wasn’t till decades later I learned the significance of the 174 Silence as a way of knowing in Yolngu Indigenous Australian storytelling names and the relationships which they implied). Wandjuk told me three stories
from his country, a thousand miles to the east in Arnhemland, including one of
the Rainbow Serpent. Wandjuk was, what some anthropologists call, a ‘bridge
person’ for his people, keen to describe his culture. More than 20 years later, I was living in the bush in solitude on the south
coast of New South Wales. It was an unprecedented year for pythons. I had
never seen so many in the 25 years I had been visiting the place. Three snakes
came to visit. The people, the place, the story The first — huge — made its way up the Angophera (a native Australian tree)
outside my window, and made its way through a hole and into the ceiling. It
was to spend the winter there, no doubt attracted by the possums who inhabited
the ceiling space during the day, too. Another python, about six feet long, twined itself into the door jam and
prevented my exit from the top storey to the outside deck, and downstairs to
the bathroom. I stamped and ‘danced’ on the floor inside the door, ‘Go away! Go away! This is not your place!’ I told my not-welcome visitor. Strange echoes
resounded — of the Wagilag Sisters. Much later I realised the parallels. In one
part of the story, the sisters are singing and dancing to keep the snake from the
door of their hut. Aah! The third snake manifested when I arrived from Sydney to set up the house
for a visit by Uncle Max3 and a group of people. Uncle Max was to lead us up
Gulaga, the sacred mountain of the Yuin people on the south coast, visible from
the back cliff behind my home — known to the Yuin as ‘pregnant woman with
her feet in the sea’. (One of the sisters is pregnant in the Wagilag Sisters Story). I opened up the house and went on to the front deck. There on the threshold
was a complete python skin — a gift — a word of poetry — evoking a shedding
of skin, transformation. Snake stories invaded my dreams. Snake stories arrived in different guises. I recalled the Rainbow Serpent story Wandjuk had told me so many years
before. I went down to the garage and found my 1979 journals and re-read the
words I had written of my meeting with Wandjuk. The snake visitations also led to a long conversation with an Aboriginal
woman (not a Yolngu), at the end of which she said she thought I could tell the
story and use my good judgment about what to tell.4 That was before I visited country of the story. I wasn’t sure. It was early
days. I read a number of versions of the story, and everything I could lay my
hands on about Yolngu country and culture. This story of the rainbow serpent is a living story I cannot tell. I am obliged
to be silent on it. The people, the place, the story I only learned of the protocols surrounding this ‘big’ story 175 Negotiating the Sacred II while delving into the question of whether I could as a non-Yolngu researcher
and storyteller tell the story. In the process of living with the question, I talked
with many people. Why, I wondered, was it that I could not tell the story? After
two years of talking with many Balanda (or non-Yolngu) people who had lived
with Yolngu and worked closely with them, I found out about what may be
called, ‘protocols’, in relation to the story. Protocols 176 Silence as a way of knowing in Yolngu Indigenous Australian storytelling I was told by Sophie Creighton, an anthropologist who has worked with
Yolngu, that there are important distinctions also between the right to hear a
story, the right to tell a story, and the bestowal of a story.5 I was told by Sophie Creighton, an anthropologist who has worked with
Yolngu, that there are important distinctions also between the right to hear a
story, the right to tell a story, and the bestowal of a story.5 Peter Sutton, a South Australian linguist writes: ‘Ancestral Beings are Stories
and their Sacred Dwelling Places are Story Places.’6 When I first saw this brief
inscription on some artefacts in the Museum of Australia in Canberra, I stopped
dead in my tracks! Something happened in my body. I did not know what it
was right then. The sentence would resonate in me for a long time. What I finally discovered after some time was that my notion of someone
telling a story, was not what it was about — the Ancestors (who are Beings, not
necessarily only human beings) are the stories — and each is intimately dwelling
as Place (in country). The tellers become the Ancestors, those who body forth in
dance as Ancestors. This turns our ordinary notions upside down — and inside out — which
brings me to ‘inside’ and ‘outside’ knowledge among the Yolngu. I take this up
later. Here is, however, the version of the story which is an acceptable form; a
public or outside version. It has been carefully worded by Yolngu. Each word
is full of significance, layers of meaning, though it is framed in ways that avoid
much detail of other now unable-to-be-public versions. This version I came to
designate, a ‘fixed version’ to distinguish it from ‘living versions’. The story covers the territory of a number of clans, in particular the waterhole
named Mirarrmina on the Upper Woolen River in Central Arnhemland. The
waterhole is the home of ‘Wititj’ the Olive Python (sometimes also known as the
‘Rainbow Serpent’), one of the most powerful of ancestral beings.7 Accounts of
the story vary with context and narrative, but the following details (the text
tells us) occur in most versions.8 Protocols First, the story emerges out of the country of Arnhemland — the story track (or
songline) moves from south-east to north. It intersects with another big creation
story of another moiety. (Each story, as well as every person, every creature and
every thing in Yolngu culture, belongs to one of two complementary moieties,
either Yirridja or Dhuwa). The story is associated with three major ceremonies
for both men and women. (Sometimes ceremonies are held separately, e.g., around
menstruation and in former times, childbirth. Some major ceremonies are held
for all.) To tell it out of country makes no sense, as the story, like the serpent,
follows the route of Yolngu Ancestors as they travelled through specific country,
which includes for example, the waterhole of Mirramina, in central Arnhemland. Custodians of the storytellings are elders from five major clans. The place of
the country where a storytelling is held will determine which part of the story
will be told. It would never be told in what we as ‘whitefellas’ would assume
to be its complete form, or any form we might find in a written ‘text’. The tellings would be carefully negotiated among the clans for a ceremony,
ensuring that all clans would be agreeable — this is part of a bringing together
and making of relationships of community, through preparation and negotiating
processes. The parts of the story and the ways in which it would be told would
depend not only on these factors, but the purpose for telling (for initiation, for
funeral or another ritual or event), the people present, their ages, gender, and
kinship with others, with the country, who the teller is, the relationship with
Dreamings in the story. In addition, kinship (or relationship with all, and with
other beings and humans) is mapped by the land — gurrutu. It interweaves and
overlays everything. The story in its oral forms — which traditionally it always is, perhaps
accompanied by singing, dancing, mark-making on bodies — in a kind of
multi-strand ‘opera’ of many senses. It is fluid and changing, dynamic, just like
the Ancestors, moving through country, changing form many times —
transformations that create country and are created out of country. The same
story would never be told on two occasions. It changes. The important elements
will however, be present. The Wagilag Sisters Story — A fixed version s9 This is the version of the story I was told by a number of expert informants9
that I could tell and write. The Wagilag Sisters Story — A fixed version Two sisters, the older of whom has a child, the younger is pregnant, are
fleeing their home in the south-east. They are being followed by
clansmen. The Sisters reach Ngilipidji, or the Stone Country of the
Wagilag clan, from where they get their name. As they continue on their
travels they encounter animals, plants and country on which they confer
names, in essence bringing them into being. They decide to camp in the
rich and fertile country surrounding the waterhole at Mirarrmina. According to some versions of the narrative, one of the Sisters pollutes
the waterhole, which arouses Wititj from his sleep. The younger Sister
gives birth, further inciting the Snake. Unsuspecting, the Sisters make
camp, build a bark hut, and try to cook the food they have caught, but 177 Negotiating the Sacred II things begin to go wrong — the animals and vegetables come to life and
leap into the waterhole. Wititj emerges from the waterhole and creates
a storm cloud with lightning and thunder to wash the Sisters into the
well. The land is flooded as the first monsoon pours down its rains. Frightened, the Sisters perform dances and sing sacred songs to deter
the Python. Finally the Sisters drop in exhaustion, and Wititj is able to enter the hut
and swallow them, their children and their dog. Shortly afterwards, the great Wititj develops a terrible stomach ache. He rises into the sky above the flooded landscape, and his groans attract
the attention of other great Snakes from surrounding clan estates, who
also rise up into the sky. They talk to each other and discover they have
different names. When asked what ails him, Wititj lies about what he
has eaten, realising that he has probably eaten beings of the same moiety. Finally, the pain becomes so great that he falls to earth and vomits the
Sisters. (He retains the children, who belong to the opposite moiety, the
Yirritja). When the Sisters are brought to life again by the bites of the
stinging caterpillars, Wititj beats them with clapsticks (malirri), and
swallows them again. The Sisters’ clansmen had followed them and were asleep by the
triangular impression made in the ground by the Python’s fall. The sisters
come to them in a dream and reveal the secrets of the sacred dances and
songs they had composed in their efforts to stop the rain. ‘Outside’ and ‘inside’ knowing Almost two years after the big snake year in my south coast home I was invited
to Arnhemland as part of a Music Forum (to include a small group of around 25
Balanda (non-Yolngu) anthropologists, ethnomusicologists, and an assortment
of others like myself, an educator, with groups of Yolngu dancers and singers
presenting and speaking) — to be followed by Garma, a festival — the occasion
of Yolngu sharing their knowledge more widely with Balanda as public or
‘outside’ knowledge. Howard Morphy, a much respected anthropologist who has worked with
Yolngu for many years, says: ‘The delicate and complex relationship between
inside and outside [knowledge] provides the context for understanding the
release of knowledge to Balanda.’10 ‘Outside’ knowledge for Yolngu (according
to Morphy) is analogous to ‘inside’ knowledge which is secret and sacred. There
are layers to this knowledge — indicated by the kinds of words that are used 178 Silence as a way of knowing in Yolngu Indigenous Australian storytelling in the storytellings for example. There will be other differences of course too. The public versions will use different words, not so-called ‘power’ words. Mandawuy told us, for example, of 30 words for the turtle, all with subtle
variations and meanings for Yolngu. Early in my explorations around the Wagilag Sisters story, I spoke with Nigel
Lendon, who had curated the large exhibition of paintings representing 60 years
of bark paintings of the story. This was held in the Australian National Gallery
in 1998. As I spoke with him of the story, he stopped me suddenly. ‘You know’,
he said, ‘If I had used the word you have just used as the name of the snake in
Arnhem Land, it would have meant death!’ He told me of being stalked while
in country. Curators had spent seven years negotiating the release of the story
in a suitable form to be published in the exhibition catalogue. I felt distinctly
uncomfortable at this news. Later, I recounted this conversation to Howard Morphy. What did he think? In his understated way, he said: ‘I think that is a little exaggerated’. However,
Alan Fidock, with whom I had worked in Canberra over many years, and who
had lived and worked with Yolngu for 13 years in Milingimby, agreed that I
could not utter this name, a ‘power word’. What to be silent about, and what to speak? ‘Outside’ and ‘inside’ knowing I knew I would have to move
slowly and with sensitivity. Speaking could have certain very serious
consequences. I did not wish to offend. I was getting conflicting messages. And
women anthropologists had different ideas again about what could and could
not be spoken!11 Much later I was to learn from Howard Morphy: Power names are called publicly at important stages of ceremonies the
whole time, e.g. the thirty words for turtle are not used in mundane
contexts. And the word you refer to as not being able to use — are used
only in restricted private contexts. However very few words fit into this
category. Most powerful names are known but treated as special. The
silence or the interval between their sounding marks their significance
and creates a sense of awe in their presence as well as the aesthetic power
of their performance.12 Later, at the Music Forum at the Yothu Yindi studio on the Bay in
Arnhemland, I asked Mandawuy a question about colour and body painting. He spoke in language to others, and eventually came back with an answer. What
was the consultation I wondered? Perhaps, how to explain in terms appropriate
to my level of understanding? I learned that one principle is to show respect,
always ask if unsure, but on the other hand, only ask a question once. Later I
found when I became closer to Gulumbu, I would ask her questions and she 179 Negotiating the Sacred II would simply turn slightly away, as though she hadn’t heard, and then I would
know not to pursue this line of questioning. Gregory Bateson says: ‘Secrecy can be used as a marker to tell us that we are
approaching holy ground…’13 Bateson’s proposition is that secrecy (or the non
utterance of certain things) was essential in the epistemological process. He says
there is a ‘necessity for the sacred’: This is a very important and significant matter, that non-communication of certain sorts is needed if we are to maintain the ‘sacred’. of certain sorts is needed if we are to maintain the ‘sacred’. ‘Outside’ and ‘inside’ knowing Communication is undesirable, not because of fear, but because
communication would somehow alter the nature of the ideas.14 Communication is undesirable, not because of fear, but because
communication would somehow alter the nature of the ideas.14 ‘Communication of certain things alters the nature of the ideas.’ Silence, or
‘non-communication’ is required for the ‘sacred’, Bateson maintains. ‘Communication of certain things alters the nature of the ideas.’ Silence, or
‘non-communication’ is required for the ‘sacred’, Bateson maintains. Howard Morphy tells us more, specifically in relation to silence as a way of
knowing in relation to Yolngu knowledge: And in many respects the layering of knowledge can be thought of as a
pedagogical [or learning process] technique, though it is one that
emphasises the variability in understanding that exists at a given moment
among different members of the society. The layering is as important as
the secrecy.15 [my emphasis] According to Morphy, it is not only the nature of the ideas, but the nature of
the process for learning which renders silence significant. Both layering and
silence have pedagogical intent, are part of the learning process. ‘Inside’ and
‘outside’ knowledge are seen to be different in Yolngu terms. Outside knowledge
is freely available. (There are ‘inside’ and ‘outside’ versions of stories). ‘Inside’
knowledge is available only to certain people at certain stages, in certain
circumstances. It is revealed in phases, with pauses, with silence between, as
Morphy has indicated — a necessary part of a sacred epistemology or way of
knowing. Morphy indicates that the concepts of ‘inside’ and ‘outside’ are more than
the English concept of ‘secret’. ‘Inside’ for Yolngu is used, he says, in two
complementary senses. The first is ‘inside’ in the sense of truth or generative
power. The other is ‘inside’ in the sense of exclusive or secret. In the first, women
and men are equally included. In the second, women are excluded from certain
contexts and certain knowledge.16 The Wagilag Sisters Story seems to hold
both kinds of knowledge. This explained its being associated with both men’s
and women’s business. ‘Outside’ and ‘inside’ knowing What Morphy has not articulated is that Yolngu women also hold certain
knowledge secret from the men — certain women’s business knowledge.17 Rita
Gross (1977) makes a strong case that men’s and women’s rituals, symbols and 180 Silence as a way of knowing in Yolngu Indigenous Australian storytelling knowledges are always equal and complementary, without gendered hierarchy,
‘co-equal and co-necessary’.18 She refutes any sacred/profane dichotomy of
male/female in Aboriginal religion and argues instead that the ‘correct
interpretation is to see women as embodiments and manifestations of a different
kind of sacrality than that associated with males’. She notes that ‘women’s
experience is inaccessible to males in the absence of articulate females’. Women’s
experience has not been fully articulated from within Aboriginal cultures.19 I
would argue that this is part of an ‘unspoken story’, a story silenced or silent,
another story that has not been told; another aspect of ‘silence’. This is the
unknowable by one gender, of the other. Rita Gross goes further. She looks
deeply into the Wagilag Sisters Story to conclude that ‘women’s unique
experiences are potent metaphors in men’s religious lives’. Another important point to be remembered is that while ‘outside’ or public
or children’s versions may be simpler in form, they reflect, or are analogous
with, the ‘inside’ knowledge, though this may not be immediately evident. Helen Verran, a philosopher now working at Charles Darwin University,
says: Yolngu knowledge is the intrusion of the Dreaming into the secular. The
Dreaming is brought into the here and now by the doing of particular
things at particular times by particular people…Knowledge can only ever
be a performance of the Dreaming, a bringing to life in the here and now of
the elements of the other domain. 20 [my emphasis] ‘Dreamtime’ in Yolngu is called Wangarr, and each language group will have
a different term which is only approximated by the English term, ‘Dreamtime’. Knowing in Yolngu is not about words, or descriptions, or abstract concepts. It
is more related to being and actioning. I will return to Helen Verran’s notion of
‘performance’ indirectly, through the dancing, a little later. I could, (I was assured by Alan Fidock, Djon Mundine, Nigel Lendon, Sophie
Creighton), write of the story and tell the story which had been carefully
negotiated and published. Revelation and epiphany are ways of knowing in Yolngu culture.21 Such
‘events’ happen in silence. What is revelation? ‘Outside’ and ‘inside’ knowing It is the known being connected
to the Unknowable in such a way as to open up to all possibility. In Yolngu culture, Bir’yun is a shimmering effect (seen in finely cross-hatched
paintings) may occur through an experience of what may be termed ‘epiphany’
or ‘revelation’. Bir’yun projects a brightness that emanates from the Wangarr
Ancestral past which manifests in the present through storytelling, singing,
art-making, dancing — or from the Ancestral Beings themselves. The brightness
of bir’yun endows the painting with ‘Ancestral power’. 181 Negotiating the Sacred II This process probably has to be present in the painter in order to evoke it in
the viewers. This may echo the state that has to be present in the storyteller for
the effect to transmit to the audience during a storytelling. The process is
similarly evoked in other forms of expression and sacred art in Yolngu (dancing,
singing), where Morphy reports that there is a progressing from dark to dull,
to light and brilliant, analogous to the transformation that arises in the process. It is present in the music which builds to a final shout, for example, as well as
in the dances which sometimes build to a frenzy of dramatic and solo
performances (especially as I noted, with the male dancers). I make the parallel
of bir’yun with storytelling and my own experiences later in country. Raymattja Marika-Mununggirity, Yolngu linguist from Yirrkala, has written
of the relationship between the ‘mythic’ ancestors and the life that is currently
lived by Yolngu: Knowledge is living, and it comes from a real world, it has real life, real
events and real happenings. That’s what happens with the old ancestral
stories, we still relive that past history, we still sing it, dance and still
bring it and fit it into the present. That’s what makes the present world
a meaningful world to live in.22 Past and present exist together, ensuring a future conserving the Ancestral states
of Being, and are given meaning and significance through re-enactment. Past and present exist together, ensuring a future conserving the Ancestral states
of Being, and are given meaning and significance through re-enactment. Revelation transforms, transmits, opens up being into Being, opens up into
all possibility. Where creatures can jump out of a fire, and run and jump into a
waterhole, as in the Wagilag Sisters Story, transformation has occurred. ‘Outside’ and ‘inside’ knowing Thus the greater interest for developing a knowledge of these works
[paintings of the Wagilag Sisters Story] is in the potential for elements
to carry multiple meanings, in different contexts, conjunctions and
historical moments.24 us t e g eate
te est o deve op
g a
ow edge o t ese wo
s
[paintings of the Wagilag Sisters Story] is in the potential for elements
to carry multiple meanings, in different contexts, conjunctions and
historical moments.24 g
p
g
g
[paintings of the Wagilag Sisters Story] is in the potential for elements
to carry multiple meanings, in different contexts, conjunctions and
historical moments.24 In Yolngu culture, all is dynamic in relation to the Story or Story versions. Contiguous elements and multivalent significances are part of the storytelling. I had come to understand that I could not tell the Wagilag Sisters Story in
any way I chose. I held no authority to tell. Not telling underscores the
importance of some secrets, the mysteries that cannot be spoken. In Yolngu society, I cannot assume a right to know. I must wait, listen, until
invited into various aspects of the assembling of understanding through action,
relationship in the environment in concert with others, with creatures, through
listening, through ritual, ceremony, through the body, through dance, through
the remarks people casually offer, and in all the details of everyday life. Story is therefore history performed. And, as Ancestors enact transformation,
so contemporary Yolngu re-create or re-enact the transformations as they tell
the stories, sing the songs, do the paintings on bodies, and dance the dances of
the Ancestors. Storytelling is in this way transformative for those who participate. In Yolngu terms, a storyteller can produce a storytelling, but it is Reality that
is presented, not just a story. Let me tell you of one experience that I had at Garma. First, the landscape. We are on a vast bush site, a stringbark forest on top of the escarpment. Between the trees, the deep red earth appears. If you walk a few hundred yards
to the edge of the escarpment you can see the sea in the distance. There are five
hundred Balanda and five hundred Indigenous people (many from vast distances
across the north of Australia) camped in tents dotted in among the trees. ‘Outside’ and ‘inside’ knowing Ancestral
Beings can transform in such a way — where an experience is charged — as
well as in a myriad other ways, and Yolngu re-enacting Ancestral Beings
re-embody that transformation. In re-storing the behaviours of the Ancestral
Beings, Yolngu re-story their own being creating Being, maintaining connection
with Ancestors. Howard Morphy, in writing of the nature of Yolngu knowledge, talks of the
revelatory nature of Yolngu knowledge and its relationship with secrecy. He
uses the word ‘transmission’ in this context. Transmission of knowledge assumes
a direct experiencing, a taking in, a knowing through embodying or ‘immediate
presencing’23 of that knowing. Morphy points out that the phasing and structuring of knowledge through
secrecy and through revelatory knowing — establishes a rhythm, that allows
slow digestion or absorption, to occur (in the way that snakes digest their food). There are thresholds at each turning, structured by secrecy. Secrecy can ‘mark
division between inside and outside knowledge’. Within such a story as the Wagilag Sisters, the potential exists for many
layers of meaning in different contexts. As Nigel Lendon has pointed out — 182 Silence as a way of knowing in Yolngu Indigenous Australian storytelling there is no one meaning for the story — meanings shift in relation to context,
juxtapositionings, and a particular moment in time: The search for a singular ‘deep’ authentic meaning is doomed to failure. The search for a singular ‘deep’ authentic meaning is doomed to failure. Thus the greater interest for developing a knowledge of these works
[paintings of the Wagilag Sisters Story] is in the potential for elements
to carry multiple meanings, in different contexts, conjunctions and
historical moments.24 The search for a singular ‘deep’ authentic meaning is doomed to failure. Franca Tamisari writes: Dancing establishes relationship between people, country and ancestors
as well as between the participants in a ceremony… In Yolngu dances
‘the body speaks — directly and in its totality’ of our being-in-the-world
and being-with-others.25 Let me remind you of a part of the Wagilag Sisters story: Let me remind you of a part of the Wagilag Sisters story: Frightened, the Sisters perform dances and sing sacred songs to deter the
Python. Finally the Sisters drop in exhaustion, and Wititj is able to enter
the hut and swallow them, their children and their dog. Frightened, the Sisters perform dances and sing sacred songs to deter the
Python. Finally the Sisters drop in exhaustion, and Wititj is able to enter
the hut and swallow them, their children and their dog. The Sisters come to [their clansmen] in a dream and reveal the secrets of
the sacred dances and songs they had composed in their efforts to stop
the rain.26 [my emphasis] The Sisters come to [their clansmen] in a dream and reveal the secrets of
the sacred dances and songs they had composed in their efforts to stop
the rain.26 [my emphasis] It is the Sunday after Garma has finished, and most of the Balanda have gone. My companions from the Music Forum have gone to a motel, but I choose to
stay on at the campsite, not wanting to sleep inside if I can be in the bush. I
have tried to get an earlier flight out, unsuccessfully. I have heard today there
is to be a ceremony, but I don’t take too much notice, as I imagine it will be held
deep in the bush, away from the eyes of myself, or other Balanda. A handful of Balanda are still there at Gulkula, the Garma site, and about
300 Yolngu — gradually drifting into the bunggul 27 space. It is hot in the sun. I have been collecting tea in large tin cans for the women,
for Galarrwuy, for the women in the weaving shelter, back and forth in the heat
to the kitchen, finding cups and sugar and milk to bring for them. It seems
appropriate to tell Galarrwuy as I bring him tea, ‘Gulumbu says I am yeppa
(sister)’. He has shown us the elaborate head-dresses and armbands that will be
for the boys, woven with coloured feathers. ‘These must be special’, I say. And so, to the story… And so, to the story… ‘Outside’ and ‘inside’ knowing A large
space opens up among the trees, covered by beach sand — the shape of a long
triangle and almost as long as a football field — where the bunggul takes place,
late each afternoon. The bunggul is the ceremonial singing, dancing, storytelling
that happens each year of Garma. The ‘boss man’ of the ceremonies and the
storyteller for Garma bunggul is Galarrwuy Yunipingu. For much of Garma I have spent time sitting on the ground with the women
in the Women’s shelter, learning how to make string, making friends with the
women, telling our stories to each other, and gradually getting to know Gulumbu,
with whom I share an instant and easy rapport. 183 Negotiating the Sacred II Franca Tamisari writes: ‘Special’, he acknowledges, as he gently puts them back into a bag. Increasingly,
it is emerging, that the ceremony will take place here in the bunggul space, for
two boys from two clans, one of them Galarrwuy’s son. I go to join the women, as Gulumbu has invited me to do, sitting in the
women’s shelter near where she is tending the fire. She leaves me in the care of
her daughter, Dhambit. Gulumbu, I realise only later, is Galarrwuy’s sister. Kin
establishes obligations, responsibilities, roles, everything. I observe that she has
a leading role to play in the unfolding event. 184 Silence as a way of knowing in Yolngu Indigenous Australian storytelling The groups for singing and dancing have assembled on either side of the
ceremonial space where the evening’s bunggul for each day of Garma have, it
seems, been gradually building to this climax. In shelters28 on opposite sides of the performance space where the figures
of Ganbulapula 29 and a great funeral log stand, sit the groups of the two related
clans. To the accompaniment of yidaki and clapsticks, they sing, each clan
alternating, and the men and women of each clan also take turns to move into
the performing space to dance in the sequence occasioned by the ceremony. Eventually, I am told, the two boys from the two clans will emerge from the
bush at one side, accompanied by the older men of both clans, to be ushered
into the space for the climax of the initiation ceremony and circumcision. They
are being prepared, painted up, in Yirrkala, the Yolngu town some distance
away. I stand near Gulumbu, tired now of sitting still in one place with the women
in the shelter for most of the day, as the singing and dancing go on, and
preparations around the bunggul continue. A great turtle is being roasted in the
ground. (Gulumbu calls to me to come and look). It will later be fed to the clan
elders at the end of the ceremony. Gulumbu is tending the fire that is to be used for the ‘smoking’ of the
relatives,30 and for boiling the ‘treatment’ for the boys, following the ceremony. I get up, stiff, and beginning to feel my body mightily in need of movement
after four or five hours of sitting still. Franca Tamisari writes: I have been saying to myself to just
observe, not ask questions on this special day, to be grateful to be present (which
I certainly am) and wait. I cannot sit still any longer. My body won’t allow it. I get up and walk across to Gulumbu at the fire. Standing opposite her as she
stirs the ‘treatment’ over the fire, I begin to move my body a little as I see the
women doing in the central sandy space. I am itching to be dancing, restless from being still for so long. Maarr, which manifests itself in the footprints left behind by ancestral
beings in the landscape features, names, objects, designs, songs and
dances — is not only ancestral power, it also refers to people’s innermost
feelings of love and care, silent wishes ‘which make things happen’,
concealed desires which are not expressed but which nonetheless are
felt and met.31 [my emphasis] Maarr, which manifests itself in the footprints left behind by ancestral
beings in the landscape features, names, objects, designs, songs and
dances — is not only ancestral power, it also refers to people’s innermost
feelings of love and care, silent wishes ‘which make things happen’,
concealed desires which are not expressed but which nonetheless are
felt and met.31 [my emphasis] I have always loved to dance almost more than anything else, and it is always
hard for me to be on the periphery of any dancing, painful to be looking in,
unable to participate. I am unsure of the codes here. I have been sitting in the
shelter observing, cautious of moving or of asking questions, conscious that I
am privileged to just be here watching, conscious too, of Gulumbu’s having
made my entry possible. 185 Negotiating the Sacred II ‘Can you teach me the dance?’ I ask Gulumbu. I do know this important
principle of being in Yolngu country. Always ask. Be respectful. I have also
been cautioned by a Balanda who has worked with Indigenous people for years:
‘Don’t ask more than once’. I can never know if what I am asking is crossing an
unknown boundary. I am risking something, transgressing the boundary of
being still, being quiet, just observing. It is risky business. But I trust this
woman, her presence, her warmth. I have put on the armband she has made me. Franca Tamisari writes: I am wearing the string with possum fur inserted into it she has given me. Gulumbu looks up from her work of stirring the pot with the stringybark in
it for the boys, straightens up from her task, then turns and gestures to the far
side of the ground, ‘You go right around the outside to that other side. Keep
right behind, and do the dancing there’. With her blessing on the enterprise giving me permission and courage, I walk
carefully around the outer rim of all the people and the activity in the middle
of the sandy performance space, trying not to run in my impatience to be dancing
— to where the group of women are seated in another shelter, as they rest in
between each segment of dancing. A tremor in my body. I am moving at last —
always for me, a favoured state. — to where the group of women are seated in another shelter, as they rest in
between each segment of dancing. A tremor in my body. I am moving at last —
always for me, a favoured state. When the women of this clan get up again to dance, I kick off my shoes and
begin to mimic them, staying well back in the background from all the people. I have positioned myself where I feel myself to be solitary, inconspicuous,
unobserved. There are only Yolngu here, but their attention is focused on what
is happening in front of me, in the bunggul. I am well behind them all. I am
enclosed in concentration, focused on being present to just the dancing. I have
the dancing group of women firmly in my sights through a kind of soft focus
vision, imitating as closely as possible their every movement — allowing my
body to dance the movements, being there, simultaneously as it is happening. It is something I have learned to do — to let go and mirror the movements, be
the moving bodies there, with my body-mind, here. So it is startling for me to hear what next takes place in the sequence of
events. I have been dancing for just a few short moments alone, but in unison with
the group a little distance away in the performance space. Franca Tamisari writes: The lead dancer turns
her head, continuing to dance as she does so, and calls over her shoulder to me,
‘Come into the canoe with us!’ At the sound of her invitation, and without a nano-second’s hesitation I leap
into the performance space (it feels in many ways a great ‘leap’), keeping my
body and mind open to picking up the steps and the rhythms as they are done,
moving with them. I am alert, but relaxed, excited, keeping my peripheral vision
open, breathing into my belly and letting the body know, forgetting any thinking 186 Silence as a way of knowing in Yolngu Indigenous Australian storytelling about this strange and daunting thing I may find myself doing, if the focus
stumbles. It is my body which is the dancing, the knowing. The reality of Yolngu performance, and especially dancing, is thus one
of epiphany and transformation in which relationships with place and people
are established, lived-in and embodied by the dancers.32 [my emphasis] I am aware of the other Yolngu women dancers, though I am focusing on the
lead dancer. I am in line with the women. The lead dancer is in front of the other
dancers, closer to the singers, and seems to emit the dance calls and to direct the
dancers. I am kicking up the dust with my feet touching the earth and sending dust
rising as each foot turns in and up slightly to the inside with each step. (This is
different from the men’s ‘hitting’ the earth with their feet). The dust itself
reminds me of a liminality of body meeting the country. As the meaning of footprints can be said to reside in between, that is in
the social, political links and emotional bonds they fashion between
places and ancestral events as well as people and country, the meaning
of dancing is between the steps, between the participants of a ceremony,
the inter-subjective space of desire and compassion, love and competition,
that one enters through dancing.33 My hands follow the dancers’ movements. I bend slightly forward as they
do, and turn as they do, re-enacting and re-embodying the journey of the
Ancestors, re-storing it to the present, re-storying it in the present. I am dressed all in black (with long white hair); they are in colourful dresses,
their dark skin glistening in the sun. Franca Tamisari writes: This country, this story, these people, this
dancing, myself Balanda, here with Yolngu, now, all related, storied, in the
dance. My hands gesture the different aspects of being in the canoe. Just dancing,
just following, being with the whole group of women, allowing my body to
keep in tune with the lead dancer who gives signals for changes in the
movements. I can just do it by focusing my attention and at the same time
opening to the body and the vision of being with what the women are doing. With a sharp shout the sequence is finished and we all retire to sit down and
the women laugh and chat among themselves in language, a language I wish
deeply to understand. Corporeal connection is enacted and elaborated in the cosmology of
Yolngu people in Northeast Arnhemland through images of bodily
transformations, journeys and traces…the body as the ‘hinge’ between
self and the world underlies the idea of ancestral power and the
experiential character of Yolngu knowledge.34 [my emphasis] 187 Negotiating the Sacred II In the dance, the body provides a silent hinge between self, world, ancestral
powers — through embodiment of ancestral power and ways of doing things. I was, though I could not reflect on it at the time, stepping directly into this
embodying by being in the dance with the Yolngu women. No matter that I did
not comprehend. I was enacting the story that was the story coming in the dance. I was in the canoe35 with the women, travelling with them, in the in-between
space and time between their culture and mine, between Ancestral Past and
contemporary time at Gulkula and between that time and the writing now. Something of a liminal silent gap quality invoked this transforming — unplanned,
unforeseen, startling. My footsteps had taken me to Darwin and the meeting with Wandjuk so long
ago, to Garma, and to meeting Gulumbu, Derrkngu, Rarriwuy, Wityana,
Mandawuy, Raymattja, and other Yolngu people, and into the dancing. All of
these events were connected it seemed to me, in tracings, in footsteps on the
country, connected through a kind of silence, the in-between and ultimately in
the embodiment that occurs as dancing the country. I left Garma with the dust of that place, Gulkula, on my feet ‘We are here now; we are the voice of the serpent,’ say the Wagilag Sisters. ‘We are here now; we are the voice of the serpent,’ say the Wagilag Sisters. ‘Now I give you my ceremonies’, says the Snake, with their voice. 36 ‘Now I give you my ceremonies’, says the Snake, with their voice. 36 Franca Tamisari writes: I had traversed three kinds of silence to reach this place: the protocols of
storytelling for Yolngu, the understanding of the parallelling of inside and
outside knowing, and a culmination of stepping into and dwelling as ancestral
spirit with the Yolngu women. All three kinds of silent knowing had coalesced
and actualised in the dancing, and in the embodying of the stories in the land
and of the land that had birthed them. I left Garma with the dust of that place, Gulkula, on my feet. Endnotes 18 18 Rita Gross, 1977, ‘Menstruation and Childbirth as Ritual and Religious Experience in the Religion
of the Australian Aborigines’, Supplement Journal of the American Academy of Religion, vol. 45, no. 4,
pp. 1147-1181, p. 1147, 1150. 18 Rita Gross, 1977, ‘Menstruation and Childbirth as Ritual and Religious Experience in the Religion
of the Australian Aborigines’, Supplement Journal of the American Academy of Religion, vol. 45, no. 4,
pp. 1147-1181, p. 1147, 1150. pp. 1147-1181, p. 1147, 1150. 19 19 There are exceptions, e.g., Deborah Bird Rose and Dianne Bell’s work in Central Australia, Fiona
Magowan and Franca Tamisari in Arnhem Land, Catherine Berndt. 19 There are exceptions, e.g., Deborah Bird Rose and Dianne Bell’s work in Central Australia, Fiona
Magowan and Franca Tamisari in Arnhem Land, Catherine Berndt. 20 H. Verran, 2002, ‘Discussion of the Philosophical Issues for Sciences and Scientists engaging with
Indigenous Knowledge’, Unpublished paper from talk, Garma (my emphasis). 20 H. Verran, 2002, ‘Discussion of the Philosophical Issues for Sciences and Scientists engaging with
Indigenous Knowledge’, Unpublished paper from talk, Garma (my emphasis). 21 21 H. Morphy, 1989, ‘From Dull to Brilliant: the Aesthetics of Spiritual Power among the Yolngu’, Man,
(N.S.), vol. 24, p. 28. 21 H. Morphy, 1989, ‘From Dull to Brilliant: the Aesthetics of Spiritual Power among the Yolngu’, Man,
(N.S.), vol. 24, p. 28. 22 R. Marika-Mununggiritj, 1991, ‘How can Balanda learn about the Aboriginal World?’ Batchelor
Journal of Aboriginal Education, July, no.6, pp. 17-23, p. 24. 22 R. Marika-Mununggiritj, 1991, ‘How can Balanda learn about the Aboriginal World?’ Batchelor
Journal of Aboriginal Education, July, no.6, pp. 17-23, p. 24. 23 Dr Tony Swain (Sydney University), expert in both Indigenous and Chinese religions and ar
maintains that there is no ‘transcendent’ in Aboriginal religions. 23 Dr Tony Swain (Sydney University), expert in both Indigenous and Chinese religions and art,
maintains that there is no ‘transcendent’ in Aboriginal religions. 24 Caruana and Lendon, op. cit. 24 Caruana and Lendon, op. cit. 25 Franca Tamisari, 2000, ‘Dancing the Land, the Land Dances through us’, Writings on Dance, no. 20,
p. 31. 25 Franca Tamisari, 2000, ‘Dancing the Land, the Land Dances through us’, Writings on Dance, no. 20,
p. 31. 26 The Wagilag Sisters Story, version in Caruana and Lendon, op. cit., p. 9. 26 The Wagilag Sisters Story, version in Caruana and Lendon, op. cit., p. 9. 36 The Wagilag Sisters Story, version Lenore, Mirelle, Balanda poet who lived in the Northern Territory
for a time with Indigenous people. She was unable to locate the sources she used for her more extensive
poem when I phoned her in Adelaide. Endnotes 1 At a conference on ‘Writing the Sacred’, The Australian National University, October 2002. 2 Commonly called in the southern areas of Australia, ‘didgeridoo’ — a long hollow log instrument
with a deep wind sound. 1 At a conference on ‘Writing the Sacred’, The Australian National University, October 2002. 2 At a conference on Writing the Sacred , The Australian National University, October 2002. 2 Commonly called in the southern areas of Australia, ‘didgeridoo’ — a long hollow log instrument
with a deep wind sound. p
3 Dulumunmun, Yuin Elder of the South Coast of New South Wales. 4 From Tranby Aboriginal College in Sydney. 4 From Tranby Aboriginal College in Sydney. 5 I could find no written source for this information. 5 I could find no written source for this information. 6 P. Sutton, 1988, Dreamings, The Art of Aboriginal Australia, New York: Viking/Penguin, p. 19 (my
emphasis). Sutton is writing of Cape York, but the same idea applies for Arnhemland. 7 W. Caruana, and N. Lendon, 1997, The Painters of the Wagilag Sisters Story 1937-1997, Canberra:
National Gallery of Australia, p. 9. 9 Nigel Lendon, Djon Mundine, Alan Fidock, Allan Marrett, Sophie Creighton — experts on different
aspects of Aboriginal and Yolngu culture. 10 H. Morphy, 1991, Ancestral Connections: Art and an Aboriginal System of Knowledge, Chicago &
London: University of Chicago Press, p. 98. 188 Silence as a way of knowing in Yolngu Indigenous Australian storytelling 11 Two Canberra-based women anthropologists told me that as the story was in the public domain I
could tell it. (They had not however, worked specifically with Yolngu, but with other Indigenous
Aborigines.) (
Aborigines.) 12 Personal email, May 2006. 12 Personal email, May 2006. y
13 G. Bateson, and C. M. Bateson, 1987, Angels Fear: Towards an Epistemology of the Sacred, New York:
Macmillan, p. 97. 14 13 G. Bateson, and C. M. Bateson, 1987, Angels Fear: Towards an Epistemology of the Sacred, New York:
Macmillan, p. 97. 1 13 G. Bateson, and C. M. Bateson, 1987, Angels Fear: Towards an Epistemology of the Sacred, New York:
Macmillan, p. 97. 14 Ibid., p. 97. 15 Morphy, 1991, op. cit., p. 77. 16 Ibid., p. 96. 17 Gulumbu Yunipingu, personal communication, August 2004. 18 17 Gulumbu Yunipingu, personal communication, August 2004. p
35 The canoe is part of the Djang’kau Sisters story, not the Wagilag Sisters, but it has an in-between,
liminal quality which seemed part of my experience at the time.
36
h
l
ll
l
d
h l
d
h
h p
35 The canoe is part of the Djang’kau Sisters story, not the Wagilag Sisters, but it has an in-between,
liminal quality which seemed part of my experience at the time.
36 The Wagilag Sisters Story, version Lenore, Mirelle, Balanda poet who lived in the Northern Territory
for a time with Indigenous people. She was unable to locate the sources she used for her more extensive
poem when I phoned her in Adelaide Endnotes 27 From bon and bun’kumu kneecap or knee; the knees of the performer moving up and down in the
dance stepping are said to be talking (bonwanga). Tamisari, op. cit., p. 35. 28 28 These are wood and bark-covered areas with sand on the floor. 28 These are wood and bark-covered areas with sand on the floor. 29 29 Ancestral figure associated with re-configuring of knowledge. 29 Ancestral figure associated with re-configuring of knowledge. 29 Ancestral figure associated with re-configuring of knowledge. g
30 Or cleansing. 31 31 Tamisari, op. cit., p. 39. 32 Ibid., p. 33. 33 Ibid. 34 Ibid., p. 3. 35 The canoe is part of the Djang’kau Sisters story, not the Wagilag Sisters, but it has an in-between,
liminal quality which seemed part of my experience at the time. 36 The Wagilag Sisters Story, version Lenore, Mirelle, Balanda poet who lived in the Northern Territory
for a time with Indigenous people. She was unable to locate the sources she used for her more extensive
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26-30, 39-42, 67, 127-128 blasphemy law
Blasphemy Act of 1698, 23
common law, 17
England, 2, 4
evolution of the charge of blasphemy
in England, 9
Quebec, 89 Abu-Sahlieh, Aldeeb, 163 Adelaide Arts Festival, 10, 37-38, 45 Adenauer
period, 69
politics of rationalism, 77 g
evolution of the charge of blasphemy Adorno, 78 Quebec, 89 Ahauutas, 37 Boucher, Denise, 83-90 Ahmadinejad, Mahmoud, 33, 142n Breakwell, Ian, 151-154, 157-158 Aikenhead, Thomas, 128, 141n Bright, Bill Rev., 97 Akhenaten, 14 British Board of Film Classification (BBFC),
3, 16 Alexander, 1 American Family Association, 96, 148 British Broadcasting Association, 97 American Family Organisation, 97 British National Viewers and Listeners
Association, 139 Ancestral Beings, 177, 181-182, 185 Ancestral Beings, 177, 181-182, 185
anaame, 108 g
anaame, 108 anaame, 108 Brown, Alan, 83 Aquinas, Thomas, 29, 39, 42 Buddha Shakyamuni, 10, 55-60, 21 Archdiocese of Montreal, 82 Bulgakov, Mikhail, 67, 69-72, 75 Arendt, Hannah, 73 Bush, George Jr., 78, 98, 100 Arthashastra, 108-109, 111, 115 Bynum Walker, Carolyn, 155, 157-158 Bynum Walker, Carolyn, 155, 157-158 Arts Council of Montreal, 82 Asprey Jacques Gallery, 150 139 Danto, Arthur, 58 Francis of Assisi, St, 155
freedom of expression, 2-4, 6, 7, 16, 23, Darabi, Louzla, 1 Francis of Assisi, St, 155
freedom of expression, 2-4, 6, 7, 16, 23,
103, 105-107, 111, 113, 117, 119, 106,
122, 131, 147
development, 12-15
freedom of religion, 103
freedom of speech, 69, 103, 122
freedom of thought, 116
Freethinker, The, 128-129, 139
Freud, Sigmund, 81
fundamentalism, 4-5, 76, 103, 105, 107, Dasaam Granth, 108 De Beauvoir, Simone, 82 103, 105-107, 111, 113, 117, 119, 106, De Moncheaux, Cathy, 155-157 Deep Faith, 151-152, 154, 158 Deep Faith, 151-152, 154, 158 Defoe, Daniel, 25, 32 freedom of religion, 103 Defoe, Daniel, 25, 32 freedom of religion, 103 Della Francesca, Piero, 149 freedom of speech, 69, 103, 122 Dening, Greg, 173 Dening, Greg, 173 freedom of thought, 116 Derrida, Jacques, 18, 95, 98-100, 121 Freethinker, The, 128-129, 139 Devoir, 82, 88 Devoir, 82, 88 Freud, Sigmund, 81 dharma, 111, 112, 118 fundamentalism, 4-5, 76, 103, 105, 107, 111, 145, 148 Dharmasashtra, 108, 110 111, 145, 148
Christian fundamentalism, 95-97, 100, Dhuwa, 176 Christian fundamentalism, 95-97, 100, Dickie, George, 151 Dionysius of Fourna, 45 Dionysius of Fourna, 45 Galavaris, George, 43-45 Galavaris, George, 43-45 dissenting voice, 6, 90 dissenting voice, 6, 90 Divine Body, 152 Ganesh (Ganesha), 37, 76, 77, 111, 117 dogma of the virgin birth/ virgin mother,
87-88 dogma of the virgin birth/ virgin mother,
87-88 Garcia Marquez, Gabriel, 78 Garma, 178, 183-184, 185, 188 Don Quixote, 28 Don Quixote, 28 Garneau, Jean-François, 88 Douglas, Mary, 41-42, 47-48, 50 Douglas, Mary, 41-42, 47-48, 50 Gaskell, Ivan, 61 Dostoevsky, Fyodor, 74 Gay News case, 2, 23, 139 Dostoevsky, Fyodor, 74 Gibson, Mel, 84, 89 Goodman, Nelson, 46, 50 Goodman, Nelson, 46, 50 Dreamings, 174, 176 Granton Star Cause, 19 Dreamtime, 173, 181 Wangarr (Yolngu), 181 Grass, Gunter, 67, 69-78 Greek Orthodox Church, 37, 42, 44, 45, 46 Greek Orthodox Church, 37, 42, 44, 45, 46 Dryden, John, 30 D’Urfey, Thomas, 28 Greenblatt, Stephen, 61 Grégoire, Paul (Mgr), 82 Emin, Tracey, 19
Enlightenment, 13-18, 75, 78, 117
European Court of Human Rights, 3
European Union Law, 16
Farohar, the, 1-2
Fatwa, 68, 75
Feinberg, Joel, 5
feminism, 81
Festival de Fringe, 83
Fidock, Alan, 178, 181
Final Solution, 71
Fingesten, Peter, 57 Gross, Rita, 180-181 Enlightenment, 13-18, 75, 78, 117 Guattari, Félix, 78 European Court of Human Rights, 3 Gulumbu, 179, 183-186, 188 Gurrutu, 176 Hale, Matthew (Lord Chief Justice), 128 Campus Crusade for Christ, 97 Campus Crusade for Christ, 97 Association of Theatre Directors, 82 Caputo, John, 99 Caputo, John, 99 Australian Constitution (section 116), 149 Caputo, Roland, 94
carnivalesque, 70-74, 88 Australian National Gallery, 1, 179 Australian National Gallery, 1, 179 carnivalesque, 70-74, 88 Carrier, Micheline, 88 cartoons,1, 129, 135, 139 Bal, Mieke, 156 Casey, Damian, 157 Balanda, 176, 178, 183-187 Balanda, 176, 178, 183-187 Caslon Analytics, 4 Baran, 162, 163, 165-170 Baran, 162, 163, 165-170 censorship, 115, 159 Barcillon, Pinin Brambilla, 59 Censorship Act of 1876 (India), 115 Barthes, Roland, 83 Iran, 161, 162, 165, 167-168, 170 Bateson, Gregory, 180 Soviet Union, 72, 74 Bazin, André, 157 Quebec, 81, 89 Bedford, Arthur, 25-26, 29-32 Bedford, Arthur, 25-26, 29-32 Clement, Sophie, 88 Behzti, 2, 107, 117-120, 122-123 Behzti, 2, 107, 117-120, 122-123 Clifford, James, 37, 48 Berger, Peter, 13 f
Colas, Emile, 82 Bersianik, Louky, 81 Coleridge, Samuel Taylor, 128-129, 131 Bharatya Janata Party (BJP), 113-114 Collier, Jeremy, 24-33 Bhatti, Gurpreet Kaur, 2, 118 Colonialism, 67, 77 Bible, 16, 28, 59, 81, 97, 99, 129, 135, 137 Communist Revolution, 75 Bildungsroman, 74, 78 Companions of Saint Laurent, 88 Birmingham Repertory Theatre, 2, 118,
122 Birmingham Repertory Theatre, 2, 118, Birmingham Repertory Theatre, 2, 118, Cordner, Michael, 28 Cordner, Michael, 28 Council of Nicea, 43 Council of Nicea, 43 Bir’yun, 181-2 Council for the Status of Women, 89 Council for the Status of Women, 89 205 Negotiating the Sacred II Fischl, Eric, 152
Fischer, Michael, 163
Fisher, Anthony, 39, 40, 41
Foote, G.W., 17, 128-129, 135, 136, 137, Cranach, Lucas, 155 Fischl, Eric, 152 Fischer, Michael, 163 Creighton, Sophie, 177, 181 Fisher, Anthony, 39, 40, 41 D’Amato, Alphonse, 148 Foote, G.W., 17, 128-129, 135, 136, 137, 97, 101, 200 iconoclasm, 1, 13, 14
artistic iconoclasm, 90
feminism, 82-87, 90 iconoclasm, 1, 13, 14
artistic iconoclasm, 90
feminism, 82-87, 90 Lawton, David, 67-68, 81, 88, 90-91, 198 legal moralism, 3-5 legal moralism, 3-5 Independent Television Commission, 97 Legault, Emile (Fr), 88, 91 Lemon, Denis, 2 Indic, 6
civilisation, 105-13, 115-17, 119, 120,
121-22
model of tolerance, 103
International Institute of Theatre, 82 Indic, 6
civilisation, 105-13, 115-17, 119, 120,
121-22
d l f t l
103 Lendon, Nigel, 179, 181-182, 188-189, 192 Les fées ont soif, v, 68, 81-90, 91, 192 model of tolerance, 103 Levinas, Emmanuel, 95, 98-99, 101, 193, Levinas, Emmanuel, 95, 98 99, 101, 193,
198 International Institute of Theatre, 82 Levy, Leonard, 21, 87, 91, 198 Iroquois Confederacy (Grand Council of
Hedenosaunee), 38, 48 Lewy, Gunter, 71, 79, 198 Lewy, Gunter, 71, 79, 198
liberal self-expression, 1
lila, 107, 109, 117
liminal
liminal space, 7
liminality, 75, 187 Iroquois
face masks, 38
people, 38 liberal self-expression, 1 liminal
liminal space, 7
liminality, 75, 187 liminal
liminal space, 7
liminality, 75, 187 Irving, David, 142 Irving, John, 69, 78 Irving, John, 69, 78 Lotsava, Gos, 56, 63 Lotsava, Gos, 56, 63 Islamic Declaration of Human rights, 163 Islamic Declaration of Human rights, 163 Love that Dares to Speak its Name, The, 2,
197 Love that Dares to Speak its Name, The, 2, Jerry Springer the Opera, 20, 23, 97 Hale, Matthew (Lord Chief Justice), 128 Harper, Davis (Hon. Justice), 4, 39, 148 heteroglossia, 69, 74 heteroglossia, 69, 74 Hoffman, Frank, J, 40-41 f
Holocaust, 71-72, 131, 142 Holocaust, 71-72, 131, 142 Holy Virgin (Mary), 38, 42, 45, 46, 50, 68,
71, 72, 82, 83, 84, 86, 88-90, 94, 147,
149, 157 Holy Virgin (Mary), 38, 42, 45, 46, 50, 68,
71, 72, 82, 83, 84, 86, 88-90, 94, 147,
149, 157 206 Index Index Koran (Quran), 1-2, 137, 151-153, 157,
163-64 Holy Virgin Mary, The, 1 Horkheimer, Max, 78 Kovats, Tania, 19, 145, 147, 149-150, 157 Human Rights League, 82 Krauss, Rosalind, 157 humanising the Divine, 68 , 84, 89, 97-98 Krishna, Kant, 112-113 Hunter, Jefferey, 94 Hunter, Jefferey, 94 Hunter, Michael, 141 Kristeva, Julia, 82-83, 154-55 Huntington, John, 61 La Presse, 82, 88, 91, 200 Huston, Nancy, 89 Lacan, Jacques, 81 hybridity, 76 Lakoff, George, 168, 171 icon, symbol and representation, 1, 37, 38
39, 42, 43, 44-47, 50, 57, 58, 63, 68, icon, symbol and representation, 1, 37, 38, Lamarche, Thérèse, 82 Last Supper, The, 10, 55-60, 62, 64 39, 42, 43, 44-47, 50, 57, 58, 63, 68, 39, 42, 43, 44-47, 50, 57, 58, 63, 68, 77, 119 Last Temptation of Christ, The, 20, 89, 93-
97, 101, 200 Jerry Springer the Opera, 20, 23, 97 Ludden, David, 113, 124, 198 Jesus (Christ), 2, 3, 4, 17, 20, 39, 40, 42-
43, 56, 58, 59, 63, 67-68, 70, 71-74, 82-
84, 89, 93-97, 98-100, 111, 129, 139,
150 52 155 Jesus (Christ), 2, 3, 4, 17, 20, 39, 40, 42-
43, 56, 58, 59, 63, 67-68, 70, 71-74, 82
84, 89, 93-97, 98-100, 111, 129, 139,
150-52, 155
Johnson, Mark, 168
Jones, Peter, 103 Macbeth, 28, 35, 191, 195 84, 89, 93-97, 98-100, 111, 129, 139, 150-52, 155 Madonna (artiste), 19 Madonna (artiste), 19 Johnson, Mark, 168 Madonna, 58, 88, 149 Madonna della Misericordia, 149 Jones, Peter, 103 Jyllands-Posten, 1 Madonna with a piano accordion, v, 10, Madonna with a piano accordion, v, 10,
37, 42, 45, 51 Kant, Emmanuel, 99 magic realism, v, 69, 73-74, 77-78 Kama Sutras, 108, 111 Kama Sutras, 108, 111 Magny, Michèle, 88 Kautaliya, 108-109 Kautaliya, 108-109 Majidi, Majid, vi, 161-169, 171 Kazantzakis, Nikos, 84, 94 Kazantzakis, Nikos, 84, 94 Malraux, André, 58, 64, 198 Malraux, André, 58, 64, 198 Keen, Ian, 146 Keen, Ian, 146 Mandawuy, 179, 188 Mandawuy, 179, 188 Khomeini, Ayatollah, 75, 162, 165 Khomeini, Ayatollah, 75, 162, 165 Maori, 46 Marcuse, Herbert, 8, 67-68, 198 Kirkup, James, 2, 139 207 Negotiating the Sacred II 149 149 Museum of Modern Art, New York, 1 power names, 179 privatisation of religion, 16 Naficy, Hamid, 161, 170, 199 pro-fanum, 70, 78 i
National Endowment for the Arts, 148 Pux, Ingrid, 90 National Federation for Decency, 96 National Museum of Canada, 37 Quebecois identity, 81 Quebecois identity, 81 Quebec State Council of the Knights of Natyashastra,
text, 105, 108-109, 111, 114-116, 119
principles
Artha, 114-116, 119, 122
Rasa, 114-116, 119, 120, 122
Abhinaya, 114-115, 119, 122
Sahmita, 114-115 Quebec State Council of the Knights of
Columbus, 82 Quebecois Writers’ Union, 82 Querelle de Tartuffe, 82, 91 Rasa, 114-116, 119, 120, 122 Quiet Revolution, Quebec, 68, 81, 83 Abhinaya, 114-115, 119, 122 Rabelais, 70, 72-74, 79, 191 Nyaya school of philosophy, 107, 109, 123 Marika, Raymattja, 182, 189, 198 Marika, Wandjuk, 174-175, 188 Observator, The, 25, 28, 35-36, 202 Martin, Jean-Hubert, 60, 64, 198 Mayal, Rik, 19 Ofili, Chris, 1 Mayal, Rik, 19 Olson, David, 97, 101, 199 McFarlane, Kyla, 148-149, 151, 153, 159,
198 Pal, Pratapaditya, 61, 63-64, 193 Menippean
satire, 72, 78, 79, 191
vision, 76 Paramount, 94, 96 Passion of Christ, The, 89 Pell v/s Council of Trustees of the National Messian, Olivier, 153 Pell v/s Council of Trustees of the National
Gallery of Victoria, 4 Gallery of Victoria, 4 Michelet, Jules, 83 Pell, George (archbishop of Melbourne),
148 Milk, Blood, 155 Mill, John Stuart, 99, 103, 129-131,135, Perret, Roy, 39-41, 52, 199 Perret, Roy, 39-41, 52, 199 Natyadharma, 114-115, 120 Rainbow Serpent (Olive Python, Wititj),
174-175, 177 Nazism
movement, 67, 75, 77
ideology, 67, 69, 74, 77
period, 67, 69-71, 75
Negri, Antonio, 117, 124, 196
Nietzsche, Friedrich, 13, 74, 76, 98, 101, Ramsey, Hayden, 39-41, 52, 195 Reformation, 13, 15-16, 21, 130, 141, 193,
196, 200 Relapse, The, 27, 35
religious pluralism, 103, 139 religious pluralism, 103, 139 Platonic understanding of art, 43-45 Mounce, Howard, 49 Plotinus, 43 Mundine, Djon, 181, 188 post-revolution Iranian cinema, 145, 161- post-revolution Iranian cinema, 145, 161- multiculturalism, 103 162, 165-167, 169 Museum of Contemporary Art, Sydney, Potter, Dennis, 20 Powell, Robert, 94 Powell, Robert, 94 Powell, Robert, 94 National Gallery of Victoria, 4, 38, 148,
150 National Gallery of Victoria, 4, 38, 148,
150 National Gallery of Victoria, 4, 38, 148,
150 141-142, 200-201 Mirarrmina, 177 Phillips, Dewi Zephaniah, 46, 49-50, 53 Phillips, Dewi Zephaniah, 46, 49-50, 53 Mockus, Michael, 17
monologism, 69, 74 Mockus, Michael, 17 picaresque, 71, 75-76, 78 Pictura Britannica, 149-150 monologism, 69, 74 Morgan, David, 62, 65, 198 Piss Christ, 4, 38-39, 41, 129, 137, 140,
147-148, 150-155, 157-159, 193 Piss Christ, 4, 38-39, 41, 129, 137, 140,
147-148, 150-155, 157-159, 193 Piss Christ, 4, 38-39, 41, 129, 137, 140, 147-148, 150-155, 157-159, 193 Morphy, Howard, 176-180, 182, 188-189,
199 Platonic understanding of art, 43-45 Platonic understanding of art, 43-45 Society for the Promotion of Christian
Knowledge (SPCK), 25, 33 Visions of Ecstasy, 2-3, 17, 204
Von Sydow, Max, 94 Von Sydow, Max, 94 Von Sydow, Max, 94 Sollers, Phillippe, 82 Sollers, Phillippe, 82 Nietzsche, Friedrich, 13, 74, 76, 98, 101, 208 199 Index Renaissance, 21, 34, 58, 64, 70, 159, 193,
195-196, 201 Tamisari Franca, 184, 189, 202 Te Papa Tongerawa Museum, 1, 145, 149-
150, 159 Rheims, Bettina, 19 Telstra, 37 Riefenstahl, Leni, 71 Telstra, 37 Riley, Jonathan, 129, 141, 200 Tempest, The, 25, 27-28, 34-35, 191, 194 Tempest, The, 25, 27-28, 34-35, 191, 194 Teresa of Avila, St, 3 Roche, Denis, 82 Teresa of Avila, St, 3 Rochefort, Christiane, 82 Tertullian, Quintus Septimus, 29-30, 35-
36, 202 Rosenbaum, Jonathan, 95, 97, 101, 200 Théatre du Nouveau Monde, 82 Roux, Jean-Louis, 82 Theotokos, 42 Theotokos, 42 Roy, Michel, 82, 88 Rushdie, Salman, 21, 67-69, 75-79, 129-
130, 136-138, 142, 191, 196-197, 200 Third Reich, 69-70 Rushdie, Salman, 21, 67-69, 75-79, 129- 130, 136-138, 142, 191, 196-197, 200 Thorsen, Jurgen, 17 Rushdie Affair, 3, 17, 34, 142, 200 Tillotson, John, 9, 26, 34, 202 Toren, Christina, 59, 64, 202 sacred epistemology, 173, 180 sacred epistemology, 173, 180 Tournier, Michel, 78 Turner Victor, 75, 79, 191 Samkhya school of philosophy, 109, 111 Tutchin, John, 25, 28-32, 35-36, 202 Santoro-Brienza, 43-44, 52, 201 Tutchin, John, 25, 28-32, 35-36, 202 Tutchin, John, 25, 28-32, 35-36, 202 Satanic Verses, The, 2-3, 7, 68, 75, 77, 129,
136, 138, 202 Universal (studio), 96
Unsworth, Clive, 2-3, 7, 202
Upanishads, the, 113
Updike, John, 137 Universal (studio), 96 Satguru, 108 Unsworth, Clive, 2-3, 7, 202 Unsworth, Clive, 2-3, 7, 202 Saur, Eberhard, 13-14 Upanishads, the, 113 Scène d’Amour, 1 Updike, John, 137 Scorsese, Martin, 20-21, 84, 89, 93-96,
100-101, 193, 195, 197, 202 Vaisesika school of philosophy, 107, 123
Van Gogh, Theo, 1
Vanbrugh, John, 27, 30, 35-36, 202
Varldskulturmuséet (Museum of World
Culture), Goteberg, Sweden, 1
Vedanta school of philosophy, 107, 109
Verran, Helen, 181, 189, 204
vilification, 2-3, 5, 112, 105
Virgin in a Condom, 1, 145, 147, 149-150,
153, 157-159, 203
Virgin Mary, 1, 38, 50, 71-72, 83-84, 147,
149, 157
Visions of Ecstasy, 2-3, 17, 204
Von Sydow, Max, 94 Vaisesika school of philosophy, 107, 123
Van Gogh, Theo, 1
Vanbrugh, John, 27, 30, 35-36, 202
Varldskulturmuséet (Museum of World
Culture), Goteberg, Sweden, 1
Vedanta school of philosophy, 107, 109
Verran, Helen, 181, 189, 204
vilification, 2-3, 5, 112, 105
Virgin in a Condom, 1, 145, 147, 149-150,
153, 157-159, 203
Virgin Mary, 1, 38, 50, 71-72, 83-84, 147,
149, 157
Visions of Ecstasy, 2-3, 17, 204
Von Sydow, Max, 94 Searle, John, 49, 53 secularisation theory, 9, 15-16, 18, 21 Sensation, 1 Sensation, 1 Sermon, The, 153-154, 157 Serrano, Andres, 4, 19, 38-39, 41, 52, 140,
143, 145, 147-148, 150-152, 154-155,
157, 159, 191, 193 Serrano, Andres, 4, 19, 38-39, 41, 52, 140,
143, 145, 147-148, 150-152, 154-155,
157, 159, 191, 193
Sex Life of Christ, 17
Shari’a law, 162
Smithsonian Museum, 37 Sex Life of Christ, 17 Virgin in a Condom, 1, 145, 147, 149-150,
153, 157-159, 203 Shari’a law, 162 Smithsonian Museum, 37 Virgin Mary, 1, 38, 50, 71-72, 83-84, 147,
149, 157 Virgin Mary, 1, 38, 50, 71-72, 83-84, 147,
149, 157 Sobchack, Vivian, 168-169, 171, 201 Society for the Promotion of Christian
Knowledge (SPCK), 25, 33 Visions of Ecstasy, 2-3, 17, 204 Visions of Ecstasy, 2-3, 17, 204 Wandjina, 37 Son of Man, 20 Sri Guru Granth Sahib, 108, 112, 114, 120-
124 Sri Guru Granth Sahib, 108, 112, 114, 120- Wagilag Sisters story, 146, 174-175, 177,
179-184, 188-189, 192 124 Wangga, the, 173 Wangga, the, 173 Sta Maria della Grazie, 56 Sta Maria della Grazie, 56 Stalinism, 67, 70, 72-74 Webster, Richard, 2-4, 7, 52, 202 Stalinism, 67, 70, 72-74 Wedde, Ian, 150 Steinberg, Leo, 155, 159, 201 Welsh, Irving, 19 Stone, Oliver, 1 Submission, 1 West, Cornel, 74, 203 Whitehouse, Mary, 2-3, 17, 19, 139, 142 Supreme Court of Victoria, 4, 39 209 Wildmon, Donald, 96-97, 101, 203 Sutton, Peter, 177, 188, 202 Negotiating the Sacred II Williams, Aubrey, 26, 34, 203 Williams, Aubrey, 26, 34, 203 Wingrove, Nigel, 3, 16-17, 21, 204 Wingrove, Nigel, 3, 16-17, 21, 204 Wittgenstein, Ludwig, 46-47, 49, 53 Wittgenstein, Ludwig, 46-47, 49, 53 Yarrow, Robin, 115, 120, 123-125, 203 Yarrow, Robin, 115, 120, 123-125, 203 Yirritja, the, 178 Yirritja, the, 178 Yolngu society, 146, 173-189, 199 Yolngu society, 146, 173-189, 199 Yothu Yindi Studio, 179 Yothu Yindi Studio, 179 Young Canadians for Christ, 82 Young Canadians for Christ, 82 Yuin people, 175, 188 Yuin people, 175, 188 Yunipingu, Galarrwuy, 183 Yunipingu, Galarrwuy, 183 Zoroastrian, 1 Zuni people, 37-39, 42, 45, 48 Zuni people, 37-39, 42, 45, 48 210 210
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Correction to: C3aR signaling and gliosis in response to neurodevelopmental damage in the cerebellum
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Kevin G. Young
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Correction to: C3aR signaling and gliosis in
response to neurodevelopmental damage
in the cerebellum Kevin G. Young1,2, Keqin Yan1 and David J. Picketts1,3,4* (2020) 17:32 (2020) 17:32 Young et al. Journal of Neuroinflammation
https://doi.org/10.1186/s12974-019-1642-x Correction to: J Neuroinflammation
https://doi.org/10.1186/s12974-019-1530-4 Following publication of the original article [1], the au-
thors noticed missing labels in Fig. 1a. The bar graph con-
tains the labels C3, C2, C1ql2, C1ql1, C1qb, GFAP, and
VGF. However, the labels should be C3, C4b, C2, C3aR1,
C1ql2, C1ql4, C1ql1, C1qa, C1qb, C1qc, GFAP, USP18,
and VGF. The correct version of Fig. 1a is published in
this Erratum. Author details
1 1Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa,
ON K1H 8L6, Canada. 2Present Address: Therapeutic Products Directorate,
Health Canada, 1600 Scott St, Ottawa, ON K1A 0K9, Canada. 3Department of
Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON K1H 8M5,
Canada. 4Department of Biochemistry, Microbiology, and Immunology,
University of Ottawa, Ottawa, ON K1H 8M5, Canada. 1.
Young KG, Yan, K, Picketts DJ. C3aR signaling and gliosis in response to
neurodevelopmental damage in the cerebellum. J Neuroinflammation.
2019;16:135. https://doi.org/10.1186/s12974-019-1530-4. References The original article can be found online at https://doi.org/10.1186/s12974-
019-1530-4 © The Author(s). 2020 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. 019 1530 4
* Correspondence: dpicketts@ohri.ca
1Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa,
ON K1H 8L6, Canada
3Department of Cellular and Molecular Medicine, University of Ottawa,
Ottawa, ON K1H 8M5, Canada
Full list of author information is available at the end of the article * Correspondence: dpicketts@ohri.ca
1Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa
ON K1H 8L6, Canada
3Department of Cellular and Molecular Medicine, University of Ottawa,
Ottawa, ON K1H 8M5, Canada
Full list of author information is available at the end of the article © The Author(s). 2020 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. © The Author(s). 2020 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Page 2 of 2 (2020) 17:32 Young et al. Journal of Neuroinflammation Young et al. Journal of Neuroinflammation (2020) 17:32 ation (2020) 17:32 Fig. 1 Altered C3 complement protein expression in the Smarca5
cKO cerebellum of exercised and sedentary mice. Increases in mR
transcripts coding for complement, complement-related proteins,
and inflammation-related proteins in the Smarca5 cKO cerebellum
as indicated by RNAseq analysis (a). Fold changes are shown for t
Smarca5 cKO groups (sedentary or exercised) relative to
corresponding wild-type groups. qRT-PCR analysis confirmed the
increases in C3, GFAP, USP18, and VGF (b), though the magnitude
of these increases varied from the RNAseq data set. Shown are th
fold changes in the Smarca5 cKO cerebellum relative to wild-type
littermates (n = 3 in each of the four groups of wild-type exercised
(run) or sedentary (sed), and mutant exercised or sedentary anima
differences relative to wild-type littermates are noted with **p <
0.005 and ***p < 0.001). No increase was observed for the C3a
receptor, C3aR. Protein analysis demonstrated a clear increase in
GFAP expression in Smarca5 cKO cerebellum samples (c). C3 prote
expression was also altered in the Smarca5 cKO cerebellum. The C
chain was less prominent relative to the C3β chain in Smarca5 cK
cerebellum samples compared to wild-type samples. Blotting resu
are representative of similar results from four mice/group Fig. 1 Altered C3 complement protein expression in the Smarca5
cKO cerebellum of exercised and sedentary mice. Increases in mRNA
transcripts coding for complement, complement-related proteins,
and inflammation-related proteins in the Smarca5 cKO cerebellum,
as indicated by RNAseq analysis (a). Fold changes are shown for the
Smarca5 cKO groups (sedentary or exercised) relative to
corresponding wild-type groups. qRT-PCR analysis confirmed the
increases in C3, GFAP, USP18, and VGF (b), though the magnitudes
of these increases varied from the RNAseq data set. Shown are the
fold changes in the Smarca5 cKO cerebellum relative to wild-type
littermates (n = 3 in each of the four groups of wild-type exercised
(run) or sedentary (sed), and mutant exercised or sedentary animals;
differences relative to wild-type littermates are noted with **p <
0.005 and ***p < 0.001). No increase was observed for the C3a
receptor, C3aR. Protein analysis demonstrated a clear increase in
GFAP expression in Smarca5 cKO cerebellum samples (c). C3 protein
expression was also altered in the Smarca5 cKO cerebellum. The C3α
chain was less prominent relative to the C3β chain in Smarca5 cKO
cerebellum samples compared to wild-type samples. Blotting results
are representative of similar results from four mice/group
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Polis, Revista de la Universidad Bolivariana, Volumen 10, Nº30 , 2011, p. 525-530
Ética de la economía
Reflexiones y propuestas de otra economía
desde América Latina
María Arcelia Gonzáles Butrón
Co-edición:
Universidad Michoacana de San Nicolás de Hidalgo y
Universidad Nacional Autónoma de México, Morelia,
2010, 274 págs.
Hugo Amador Herrera Torres
Universidad Michoacana de San Nicolás de Hidalgo, Morelia, Michoacán, México.
E-mail: hugoht@fevaq.net
El sólo título Ética de la economía, con que bautiza la Dra. Gonzáles
Butrón a su libro, resulta sugerente. El prólogo se manifiesta también sugerente. Cerutti, quien lo escribe, explica que la economía implica valores y
que estos valores determinan criterios que colocan a los seres humanos en
determinadas posiciones, donde las neutralidades son meras apariencias.
Cerutti argumenta que las posiciones que implican pisar las cabezas, los
cuerpos, las necesidades y las dignidades de los otros y las otras vienen de
valores, aunque parezca mentira. Esto importa mucho, dice Cerutti, porque
sin otras ni otros no hay nosotros ni nosotras. Pisar al otro o la otra implica
pisarse a uno mismo. Son posiciones fundadas en valores destructivos. El
mismo Cerutti expresa que la autora del libro llega a este planteamiento:
sin tú, no hay yo, que es la noción de sujeto que maneja Hinkelammert, la
idea de otredad de Foucalt y la explicación de sujeto y otredad de Morin.
La autora trata de comprobar dos hipótesis en su libro (mencionadas textualmente). Primera, que las diversas manifestaciones de exclusión
social son signos del predominio de una economía de mercado total cimentada en modelos de crecimiento económico que no ponen en el centro de
sus propuestas al ser humano sino a la acumulación de capital. Esta economía de mercado total tiene su propia ética, que determina la manera de
comportarse del ser humano, no sólo en la dimensión económica sino también en la social, política, simbólica, cultural, ideológica y religiosa. La
Dra. González Butrón aquí toma una proclama de Nietzsche: la ética revela
la verdadera intención de toda filosofía, su trasfondo oculto y a veces inconsciente. La ética no constituye una aplicación de una teoría gnoseológica
o metafísica sino “el germen verdadero de donde nace la planta completa”.
En la segunda hipótesis, plantea que resquebrajar y desmontar los
supuestos de la ética de la economía de mercado total puede repercutir en
una mayor eficacia en la crítica de esta ética y, además, puede contribuir a
525
Polis, Revista de la Universidad Bolivariana, Volumen 10, Nº 30, 2011
fertilizar los terrenos económicos que buscan transformar la ética económica imperante. La primera parte de esta hipótesis puede comprobarse con
la eficacia que alcance la misma escritora en la crítica que hace a la economía de mercado total. La segunda parte de la hipótesis no puede comprobarse con los temas tratados en el libro, sino con los efectos que tengan
estos temas en trabajos teóricos y prácticos posteriores.
La crítica que se hace en la obra no parte de la nada, tiene un punto
de partida: la preservación de la vida humana y natural. Es un libro que está
ubicado entonces en los confines del pensamiento crítico. En el contenido
de los cuatro capítulos que conforman al texto se detecta con claridad la
producción teórica de Hinkelammert.
En el primer capítulo, la Dra. Gonzáles Butrón expone que las políticas de ajuste estructural implementadas en América Latina -principalmente
durante la década de los ochenta- condujeron a cambios estructurales de
largo plazo más que a “ajustes” coyunturales de corto plazo, cambios
que beneficiaron “sólo” a las empresas transnacionales y perjudicaron a
“todos” los países latinoamericanos, sin excepción. La autora, citando a
Hinkelammert, escribe que estos cambios significaron una adaptación
de América Latina al funcionamiento del capitalismo extremo, conducido por el mercado total, buscando que los países centrales absorbieran
el máximo excedente posible -o hasta el imposible- de los países latinoamericanos.
La escritora, a manera de ilustración, hace un análisis económico
que muestra el comportamiento de la economía mexicana, toma el periodo
1980-2007. Llega a la conclusión de que la economía de México ha estado
estancada como consecuencia de la adopción de las políticas de ajuste estructural, sin hallazgos de recuperaciones momentáneas. El análisis lo desarrolla empleando variables clásicas de la síntesis neoclásica-keynesiana:
producto interno bruto, cuenta corriente, inversiones extranjeras, inflación,
pobreza con base en ingresos. Los resultados negativos los interpretó como
la insuficiencia de estas políticas. Pero tal pareciera, que si la autora hubiera encontrado resultados positivos con las variables que utilizó, estas políticas serían suficientes. La experiencia -con evidencias contundentes- ha
mostrado que el crecimiento económico no necesariamente se relaciona
con sociedades inclusivas, muchas veces el crecimiento ha venido acompañado de exclusión social. El análisis de la Dra. Gonzáles Butrón se ve limitado, requiere de la incorporación de más variables. Aunque los resultados
económicos de las políticas de ajuste estructural hubieran salido positivos,
sus consecuencias serían -de igual manera- nefastas para la mayoría de la
sociedad mexicana. Son políticas, como diría Hinkelammert, promotoras
del mercado total, su esencia es destructiva.
En el segundo capítulo, se define a la ética como una ciencia encargada de estudiar la moral, de reflexionar sobre el comportamiento moral
del ser humano en la sociedad. La autora, estudiando la mirada ética de
Hayek, describe la moral de la sociedad moderna, caracterizada por com-
526
Hugo Amador Herrera Torres
portamientos basados en el respeto a la propiedad privada y a las obligaciones contraídas (derecho contractual). Estas dos características, consideradas como valores normativos, constituyen las piedras angulares, que según Hayek, permiten al ser humano “progresar”. La moral que postula valores antagónicos a la propiedad privada y a los contratos resulta entonces
una moral inmoral.
Esta concepción de moral, dice la escritora, es una cosmovisión que
gira sobre paradigmas epistemológicos, metodológicos y praxeológicos,
que hace que los seres humanos organicen su vida como individuos aislados, interesados y calculadores (homo economicus). Esta organización social es la que impulsa la reproducción del mercado total.
En el tercer capítulo, muy vinculado con el segundo, la Dra. Gonzáles
Butrón centra el análisis en la ética del mercado total. Es mercado “total”
porque guía “todas” las acciones del ser humano (reducido a un individuo
aislado e interesado con subjetividad invisibilizada) mediante relaciones
mercantiles. Su lógica abarca “todo”. La ética del mercado total gira sobre
un valor central: la eficiencia (Hinkelammert). Si los individuos logran alcanzar sus fines específicos con la menor cantidad posible de medios, entrarán en las esferas de la productividad y, por ende, tendrán la posibilidad
de competir en el mercado. La eficiencia se transforma así en el valor supremo que decide sobre la validez de “todos” los demás valores.
La propiedad privada y el derecho contractual alientan la eficiencia,
por eso se consideran valores. Estos conceptos -o mejor dicho valores- son
la base para desarrollar una nueva personalidad, que haga posible la aparición de un entorno donde el individuo pueda alcanzar sus fines específicos
con la máxima eficiencia posible. La ética de la economía de mercado total
se sustenta en esta racionalidad, que es la racionalidad medio-fin (Max
Weber).
Es una racionalidad que impone un modus operandi basado en la
eficiencia. No obstante, pensar en términos medio-fin de manera lineal oscurece los efectos que esta misma operación produce contra la naturaleza
humana y no humana. No importan -en esta racionalidad- los medios utilizados para obtener los fines propuestos, aunque se impida la reproducción
de la misma vida. Se trata de una racionalidad irracional que no permite
criterios valorativos sobre sus hechos. Se introducen cambios pero nada
cambia en su funcionamiento básico, excepto su velocidad (Walter Benjamín).
La sociedad moderna se apunta por una serie de muertes y finales: la
muerte del sujeto, de la historia, de la razón, el fin de los relatos de emancipación, de las ideologías. Muertes todas ellas anunciadas por la proclama
nietzschiana de la “muerte de Dios”. Es el resumen y el paradigma de tantas defunciones. Pero no se trata de muertes naturales, sino de asesinatos
(Klappenbach). Estos asesinatos son las mismas pisadas a las cabezas humanas a las que se refiere Cerutti en el prólogo.
527
Polis, Revista de la Universidad Bolivariana, Volumen 10, Nº 30, 2011
En el cuarto capítulo, la escritora, tomando como soporte un trabajo
de Leonardo Boff, señala cuatro posibles vetas de alternativas a la economía de mercado total: neokeynesianismo, ecosocialismo, poscapitalismo y
Carta de la Tierra. Menciona que el neokeynesianismo es una alternativa
contradictoria, pues busca regular al mercado total sin cuestionar e interpelar sus efectos perversos, sería como pedirle al lobo que dejara de devorar
ovejas. Al ecosocialismo lo describe como una alternativa de amplias posibilidades mientras sea integral. En el poscapitalismo identifica un centro
rector: recuperar el sentido clásico y etimológico de la economía, orientarla hacia la creación de las bases materiales para el bienestar físico, cultural
y espiritual de los seres humanos. Se trata de la oikonomia de Aristóteles.
Al poscapitalismo, sin embargo, lo marca como alternativa difícilmente
realizable, está fuera de los marcos de factibilidad.
La alternativa de la Carta de la Tierra, siguiendo a Boff y a la propia
autora, se maneja como la opción con mayor grado de concretización. Esta
alternativa parte -al igual que el poscapitalismo- de una opción radical por
la vida y por la Tierra. La escritora no logra clarificar con precisión las
diferencias teóricas existentes entre poscapitalismo y Carta de la Tierra,
hace explícito que ambas giran sobre la vida humana y natural, pero señala
diferentes niveles de factibilidad, que no son profundizados.
Sin salirse de los límites de estas dos alternativas, la Dra. Gonzáles
Butrón trabaja el paradigma del principio de la equivalencia de Peters, la
propuesta de un régimen de propiedad desde abajo de Duchrow y
Hinkelammert, la economía social y solidaria centrada en el trabajo de
Coraggio, la racionalidad reproductiva de Hinkelammert y Mora, y la
mundialización de la ciudadanía como totalidad de Dierckxsens. En estas
propuestas, según la escritora, están los horizontes de una economía para la
vida, horizontes alternativos a la idea imperante de homologar economía y
mercado en busca del homo economicus, del individuo ahistórico, desconectado de su comunidad, maximizador, interesado, egoísta y competitivo.
Se trata del individuo que se mueve en la crematística (arte del lucro, del
enriquecimiento) de Aristóteles.
La ética de la economía que presenta la autora va sobre las líneas de
la ética de la responsabilidad del bien común que plantea Hinkelammert y
de la ética de la liberación que presenta Dussel. El común denominador de
estas propuestas es la reproducción de la vida humana y natural. Son propuestas donde la vida del ser humano, en todas sus dimensiones, y la preservación del entorno ecológico, aparecen como la totalidad de donde deben desprenderse los valores éticos. La racionalidad medio-fin aparece supeditada a esta totalidad.
La escritora en sus conclusiones (pág. 228) habla de resultados sociológicos que implican la muerte del sujeto. Esto entra en debate.
Hinkelammert (2004: 15-16), autor más citado en la obra, explica que el
humano al ser reducido a individuo propietario y calculador de sus utilidades, invisibiliza su otro polo, que es el sujeto, pero no desaparece, no mue-
528
Hugo Amador Herrera Torres
re, es reprimido. La autora (págs. 212 y 236) hace mención también del
otro polo del individuo, el sujeto (citando a Hinkelammert), pero no habla
sobre su invisibilidad, se queda ahí nada más. Ella misma cierra su obra
(págs. 237) asegurando que la afirmación del sujeto está emergida en la
esperanza que da vida. Ahora niega la muerte del sujeto. El sujeto es la
única instancia desde la cual se puede enfrentar al mercado total. La muerte
del sujeto implica borrar cualquier alternativa. La muerte del sujeto implica aceptar que el mercado total efectivamente no tiene alternativas.
El libro Ética de la Economía. Reflexiones y propuestas de otra
economía desde América Latina, de la Dra. González Butrón, sin duda
conduce y coloca contra las cuerdas, deja ver que no resulta fecundo seguir
como si nada, que es necesario asumir responsabilidades y aventurarnos en
la idea de que otra economía es posible (Cerutti). Quizá, el debate en que
nos sumerge hubiera sido todavía más profundo con: 1) un tratamiento teórico especial a la exacerbación de la teoría neoclásica en el neoliberalismo,
2) relacionar los valores éticos que manejan en sus obras Adam Smith y
Hayek, 3) incorporar en el debate a Amartya Sen, quien es presentado al
inicio del libro como uno de los autores que ha identificado el distanciamiento entre ética y economía, pero su propuesta no se desarrolla en el
cuarto capítulo (capítulo de alternativas al mercado total).
Ninguna obra puede considerarse concluyente y definitiva, todas
quedan abiertas. El valor teórico del libro presentado es alto, se logran
sistematizar -en buen grado- diversos pensamientos en torno a un tema común, la ética del mercado. La autora logró comprobar una de sus hipótesis
y parte de la otra, bueno, al momento que se decide hacer una reseña sobre
su libro, se revelan ya efectos producidos en los lectores. La Dra. Gonzáles
Butrón se enfrentó con buen nivel teórico al orden ético fundado en la aceptación del asesinato del otro y de la otra.
529
Polis, Revista de la Universidad Bolivariana, Volumen 10, Nº 30, 2011
Bibliografía
Hinkelammert, Franz, (2004), “La vida es más que el capital. La democracia de ciudadanos y el proyecto de la sociedad en la que quepan todos los
seres humano”, en Pasos, Departamento Ecuménico de Investigaciones,
San José de Costa Rica.
530.
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Correction to: Chest x-ray severity score in COVID-19 patients on emergency department admission: a two-centre study
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Cristian Giuseppe Monaco
|
English
|
Spoken
| 666
| 1,304
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Correction to: Chest x-ray severity score in
COVID-19 patients on emergency
department admission: a two-centre study Cristian Giuseppe Monaco1, Federico Zaottini2, Simone Schiaffino1*, Alessandro Villa2, Gianmarco Della Pepa3,
Luca Alessandro Carbonaro1, Laura Menicagli1, Andrea Cozzi4, Serena Carriero3, Francesco Arpaia3,
Giovanni Di Leo1, Davide Astengo2, Ilan Rosenberg2 and Francesco Sardanelli1,4 Author details
1 1Unit of Radiology, IRCCS Policlinico San Donato, Via Rodolfo Morandi 30,
20097 San Donato Milanese, Italy. 2Unit of Radiology, Ospedale Lavagna, Via
Don Giovanni Battista Bobbio 25, 16033 Lavagna, Italy. 3Postgraduate School
in Radiodiagnostics, Università degli Studi di Milano, Via Festa del Perdono 7,
20122 Milan, Italy. 4Department of Biomedical Sciences for Health, Università
degli Studi di Milano, Via Luigi Mangiagalli 31, 20133 Milan, Italy. Correction to: Eur Radiol Exp 4, 68 (2020)
https://doi.org/10.1186/s41747-020-00195-w The original article [1] mistakenly omitted a declaration
of competing interests. An appropriate statement can be
viewed ahead: © The Author(s). 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License,
which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give
appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if
changes were made. The images or other third party material in this article are included in the article's Creative Commons
licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons
licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain
permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Competing Interests Francesco Sardanelli is the Editor-in-Chief of European
Radiology Experimental; for this reason, he was not in-
volved in any way in the revision/decision process, which
was completely managed by the Guest Editor, Prof. Akos
Varga-Szemes (Medical University of South Carolina,
Charleston, SC, USA). * Correspondence: schiaffino.simone@gmail.com * Correspondence: schiaffino.simone@gmail.com
1Unit of Radiology, IRCCS Policlinico San Donato, Via Rodolfo Morandi 3
20097 San Donato Milanese, Italy
Full list of author information is available at the end of the article * Correspondence: schiaffino.simone@gmail.com
1Unit of Radiology, IRCCS Policlinico San Donato, Via Rodolfo Morandi 30,
20097 San Donato Milanese, Italy
Full list of author information is available at the end of the article Full list of author information is available at the end of the artic © The Author(s). 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License,
which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give
appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if
changes were made. The images or other third party material in this article are included in the article's Creative Commons
licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons
licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain
permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Reference 1. Monaco CG et al (2020) Chest x-ray severity score in COVID-19 patients on
emergency department admission: a two-centre study. Eur Radiol Exp. 4:68. https://doi.org/10.1186/s41747-020-00195-w Monaco et al. European Radiology Experimental (2021) 5:17
https://doi.org/10.1186/s41747-021-00215-3 Monaco et al. European Radiology Experimental (2021) 5:17
https://doi.org/10.1186/s41747-021-00215-3 (2021) 5:17 Monaco et al. European Radiology Experimental
https://doi.org/10.1186/s41747-021-00215-3 European Radiology
Experimental CORRECTION Open Access CORRECTION
Open Access
Correction to: Chest x-ray severity score in
COVID-19 patients on emergency
department admission: a two-centre study
Cristian Giuseppe Monaco1, Federico Zaottini2, Simone Schiaffino1*, Alessandro Villa2, Gianmarco Della Pepa3,
Luca Alessandro Carbonaro1, Laura Menicagli1, Andrea Cozzi4, Serena Carriero3, Francesco Arpaia3,
Giovanni Di Leo1, Davide Astengo2, Ilan Rosenberg2 and Francesco Sardanelli1,4 The original article can be found online at https://doi.org/10.1186/s41747-
020-00195-w. The original article can be found online at https://doi.org/10.1186/s41747-
020-00195-w.
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Transcriptomic Analysis of (Group I) Clostridium botulinum ATCC 3502 Cold Shock Response
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Elias Dahlsten
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English
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Received October 31, 2013; Accepted January 24, 2014; Published February 24, 2014 Received October 31, 2013; Accepted January 24, 2014; Published February 24, 2014 Copyright: 2014 Dahlsten et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This research was performed in the Finnish Centre of Excellence in Microbial Food Safety Research (http://www.vetmed.helsinki.fi/mifosa/) and was
funded by the Academy of Finland (http://www.aka.fi/en-GB/A/)(grants 118602, 1115133, and 1120180), the ABS Graduate School (http://www.vetmed.helsinki.fi/
abs/index.htm), the Walter Ehrstro¨m Foundation (http://www.maitohygienialiitto.fi/walter-ehrstromin-saatio), and the Finnish Foundation of Veterinary Research
(http://www.sels.fi/index_eng.htm). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: elias.dahlsten@helsinki.fi . These authors contributed equally to this work. Exposure to low temperature presents several challenges to the
bacterial cell. Upon cold shock, the translational machinery is
hampered due to formation of stable secondary mRNA structures
and decreased ribosome activity. Moreover, the folding and
activity of proteins is slowed down, and the solidification of the
cytoplasmic membrane lipids hinders biomolecule transport and
other membrane-associated processes [5]. To counter these
constraints, the cell elicits a set of targeted defensive responses,
namely, the cold-shock response. The genome-wide response to
temperature downshift has been extensively characterized in the
Gram-positive model organism Bacillus subtilis [6,7]. However,
little is known regarding machineries related to sensing and
adapting of C. botulinum to low temperature. Abstract Profound understanding of the mechanisms foodborne pathogenic bacteria utilize in adaptation to the environmental
stress they encounter during food processing and storage is of paramount importance in design of control measures. Chill
temperature is a central control measure applied in minimally processed foods; however, data on the mechanisms the
foodborne pathogen Clostridium botulinum activates upon cold stress are scarce. Transcriptomic analysis on the C. botulinum ATCC 3502 strain upon temperature downshift from 37uC to 15uC was performed to identify the cold-responsive
gene set of this organism. Significant up- or down-regulation of 16 and 11 genes, respectively, was observed 1 h after the
cold shock. At 5 h after the temperature downshift, 199 and 210 genes were up- or down-regulated, respectively. Thus, the
relatively small gene set affected initially indicated a targeted acute response to cold shock, whereas extensive metabolic
remodeling appeared to take place after prolonged exposure to cold. Genes related to fatty acid biosynthesis, oxidative
stress response, and iron uptake and storage were induced, in addition to mechanisms previously characterized as cold-
tolerance related in bacteria. Furthermore, several uncharacterized DNA-binding transcriptional regulator-encoding genes
were induced, suggesting involvement of novel regulatory mechanisms in the cold shock response of C. botulinum. The role
of such regulators, CBO0477 and CBO0558A, in cold tolerance of C. botulinum ATCC 3502 was demonstrated by deteriorated
growth of related mutants at 17uC. Citation: Dahlsten E, Isokallio M, Somervuo P, Lindstro¨m M, Korkeala H (2014) Transcriptomic Analysis of (Group I) Clostridium botulinum ATCC 3502 Cold Shock
Response. PLoS ONE 9(2): e89958. doi:10.1371/journal.pone.0089958 Editor: Michel R. Popoff, Institute Pasteur, France Received October 31, 2013; Accepted January 24, 2014; Published February 24, 2014 Identification of Cold-regulated Genes in C. botulinum
ATCC 3502 To identify the coding sequences (CDSs) that were significantly
induced or repressed upon temperature downshift in C. botulinum,
we carried out a transcriptomic analysis of the ATCC 3502 strain. The abundance of transcripts in cultures grown at 37uC were
compared to those exposed to cold stress at 15uC for 1 h or 5 h
using DNA microarrays based on the ATCC 3502 genome [12]. Genes up- or down-regulated by a log2-ratio of .2.0 or 2,2.0
(false discovery rate [FDR] ,0.05), respectively, were considered
to be cold-regulated. In total, 199 CDSs were up-regulated 5 h
after the cold shock; of these, 16 CDSs showed induction also 1 h
after temperature downshift. The analysis revealed 210 CDSs to
be down-regulated 5 h after exposure to low temperature; of these,
10 CDSs showed decreased transcript levels 1 h after the cold
shock. Additionally, one CDS (cbo2459) was down-regulated 1 h
but not 5 h after the cold shock. The cold-regulated CDSs were
equally distributed among the genome (Fig. 1). The cold-induced
and repressed gene set of C. botulinum ATCC 3502 is presented in
Table S1. To gain an insight into genes with similar transcription patterns
upon cold shock, the number of up- and down-regulated genes in
each functional category was determined. The functional category
for each gene was derived from the assignment in the original
annotation of the ATCC 3502 genome [12]. This revealed a
marked number of down-regulated genes in central metabolism-
related categories, and up-regulated genes in adaptation- and
regulation-related categories (Fig. 4). The significant components
of
these functional
categories
were further
combined into
biologically relevant metabolic groups when considered necessary. Validation of the DNA Microarray Results with RT-qPCR
Quantitative real-time reverse-transcription PCR (RT-qPCR)
analysis was carried out to validate the 1-h post-shock expression
fold change data obtained from the microarray experiments. Fold
changes of transcript levels for genes cbo0097, cbo0477, cbo0558A,
cbo0751, cbo0753, cbo1407, cbo2226, cbo2227, cbo2525, cbo2847,
cbo2961, cbo3199 and cbo3202 one hour after the cold shock,
normalized to 16S rrn transcript levels and calibrated to pre-cold-
shock transcript levels, were calculated using the Cq values
obtained from qPCR runs. The Cq values of the 1-h post-shock
transcript levels were obtained in a previous study [13] for genes
cbo0751, cbo0753, cbo1407, cbo2226, cbo2227, cbo2525, cbo2847,
cbo3199, and cbo3202; all other data were produced in the current
study. The Cold-affected Gene Set 1 h after Temperature
Downshift in C. botulinum ATCC 3502 A set of 27 genes was up- or down-regulated 1 h after
temperature downshift (Table S1). Among these was cbo2802
(deaD) putatively encoding a DEAD/DEAH box family RNA
helicase, with 4.3-fold up-regulation 1 h after the cold shock. Additionally, cbo0282 (cspA) encoding the cold-shock protein CspA
was 6.6-fold up-regulated. The genes cbo1636 and cbo1637
predicted to encode the ATP-binding protein and permease A
components of the glycine betaine/L-proline ABC transporter
OpuC, were induced 4.6- and 6.5-fold, respectively, 1 h after the
cold shock. In addition to the aforementioned annotated genes, several
genes without predicted functions were induced. Among these,
cbo2592 and cbo2591, the first genes of the putative operon
cbo2592-cbo2581, were 9.2-and 7.0-fold induced 1 h after cold
shock, respectively. Additionally, 6.5- to 8.6-fold induction of a
putative operon cbo0558A-cbo0560 encoding a DNA-binding
transcriptional regulator and components of a putative ABC
transporter was observed, as well as 16-fold induction of cbo0389
encoding a putative amino acid permease. Results Each of the four k-means clusters was separately subjected to
hierarchical clustering in order to identify sub-groups of genes with
a similar transcriptional pattern within the clusters. The aim of this
analysis was to identify putative functional associations within
small groups of similarly transcribed genes. Hierarchical clustering
grouped together some functional and/or transcriptional units,
such as genes involved in anaerobic respiration or regulation of
iron metabolism in cluster 1, or genes putatively involved in fatty
acid biosynthesis in cluster 3. However, no further association of
sub-clusters to functional categories or metabolic pathways could
be identified. The k-means main cluster and the hierarchical sub-
cluster of each differentially transcribed gene are shown in Table
S1, along with the functional main- and sub-classes for all the
genes represented in the microarray. Identification of Cold-regulated Genes in C. botulinum
ATCC 3502 In a linear regression analysis between the microarray and
RT-qPCR log2 fold changes (Fig. 2), a R2 correlation value of 0.93
was observed. Transcriptome of C. botulinum in Cold Shock Transcriptome of C. botulinum in Cold Shock To gain a more comprehensive picture of the cold-shock
response
of
C. botulinum
ATCC
3502,
we
carried
out
a
transcriptomic analysis after a temperature downshift from the
optimal 37uC to 15uC. Genes with .2.0 or ,2
22.0 log2-fold
difference in their transcript levels in comparison to the levels prior
to cold shock were considered to be cold-induced or repressed,
respectively. We identified differential transcription of 27 genes
1 h after the cold shock, whereas several hundreds of genes were
affected 5 h after the cold shock. The most abundant groups of
strongly
up-regulated
genes
were
involved
with
fatty
acid
biosynthesis, transcriptional regulation, and iron transport and
storage functions. Additionally, insertional inactivation of the up-
regulated cbo0477 and cbo0558A, putatively encoding DNA-
binding
regulators,
resulted
in
deteriorated
cold
tolerance,
suggesting roles for these regulators in the cold stress response of
C. botulinum ATCC 3502. main groups was accomplished with k-means clustering method,
which is commonly used in microarray data analysis. K-means
clustering partitions the data into a user-defined number of clusters
based on the expression values included in the analysis. From
among the different number of tested k-means clusters, the
amount of clusters was chosen to be four as this exhibited
biologically relevant partitioning of the genes primarily based on
the level of induction or repression at 5 h after the cold shock. Clusters 1 and 2 contained 156 and 167 up- and down-regulated
genes, respectively, whereas the number of up- and down-
regulated genes in clusters 3 and 4 were 43 and 44, respectively
(Fig. 3). The genes within clusters 3 and 4 exhibited markedly
stronger up- and down-regulation within the 5-hour experimental
time window, respectively, than the genes partitioned into clusters
1 and 2. As the number of differentially transcribed genes at 5 h
was substantially greater than at 1 h, the cluster allocation was
mostly affected by the 5-h transcript levels and thus failed to
notably cluster together genes only affected at 1 h. Introduction Understanding the mechanisms by which foodborne pathogenic
microorganisms cope with stress conditions they encounter in
foods is of key importance in designing modern food safety
measures. The ability of the anaerobic Gram-positive spore-
forming Clostridium botulinum to survive, grow and subsequently
produce botulinum neurotoxin in foods [1] raises substantial
concern over food safety [2,3]. The approaches to control growth of C. botulinum in minimally
processed foods differ considerably from those of canned foods,
where autoclaving to destroy 12 log-units of spores is the standard
control method. In minimal food processing, combinations of
environmental hurdles including low water activity, low or high
pH, and, most importantly, low storage temperature are used. Exposure of bacteria to sub-lethal stress can result in increased
robustness and (cross-)protection towards harsher treatments, thus
creating challenges in classical hurdle design in food processing
[4]. Hence, identification of mechanisms behind response and
adaptation to the environmental hurdles C. botulinum may
encounter in food processing might provide biomarkers for
detection of potentially stress-adapted cells, thus allowing more
precise and efficient control. Of the three cold-shock domain family proteins (Csps) present in
the C. botulinum type A strain ATCC 3502, CspB has been shown
to be the major cold-related Csp [8]. Recently, we have identified
two two-component systems (TCS), CBO0366/CBO0365 [9] and
CBO2306/CBO2307 [10], that had induced expression upon
temperature downshift and an important role in cold adaptation in
C. botulinum ATCC 3502. Furthermore, a novel role in cold and
hyperosmotic stress tolerance of C. botulinum was demonstrated for
the previously strictly sporulation-associated alternative sigma
factor SigK [11]. February 2014 | Volume 9 | Issue 2 | e89958 1 PLOS ONE | www.plosone.org Hierarchical Clustering of Differentially Transcribed
Genes and Determination of the Number of Differentially
Transcribed Genes in each Functional Category To identify groups of similar, biologically relevant transcrip-
tional patterns among the cold shock-affected genes, clustering of
the data into main- and sub-clusters was performed. Division into February 2014 | Volume 9 | Issue 2 | e89958 February 2014 | Volume 9 | Issue 2 | e89958 PLOS ONE | www.plosone.org 2 PLOS ONE | www.plosone.org Transcriptome of C. botulinum in Cold Shock Figure 1. Projection of genes significantly up- or down-regulated upon cold shock in the C. botulinum ATCC 3502 genome. From
outside to inside: Ring 1, molecular clock of C. botulinum ATCC 3502 genome; ring 2, coding DNA sequences of the forward strand (red); ring 3,
coding DNA sequences of the reverse strand (cyan); ring 4, ribosomal and transfer RNA (blue); ring 5, pseudogenes (brown) and prophages (orange);
ring 6, genes up-regulated (pink) or down-regulated (green) 1 h after the cold shock; ring 7, genes up-regulated (pink) or down-regulated (green) 5 h
after the cold shock. doi:10 1371/journal pone 0089958 g001
p Figure 1. Projection of genes significantly up- or down-regulated upon cold shock in the C. botulinum ATCC 3502 genome. From
outside to inside: Ring 1, molecular clock of C. botulinum ATCC 3502 genome; ring 2, coding DNA sequences of the forward strand (red); ring 3,
coding DNA sequences of the reverse strand (cyan); ring 4, ribosomal and transfer RNA (blue); ring 5, pseudogenes (brown) and prophages (orange);
ring 6, genes up-regulated (pink) or down-regulated (green) 1 h after the cold shock; ring 7, genes up-regulated (pink) or down-regulated (green) 5 h
after the cold shock. doi:10.1371/journal.pone.0089958.g001 putatively encodes the primary fatty acid biosynthesis machinery
of C. botulinum ATCC 3502. The Effects of Prolonged Cold Exposure on the
Transcriptome of C. botulinum ATCC 3502 Oxidative stress and metal ion homeostasis. Induction of
several genes related to countering oxidative stress and maintain-
ing redox balance, including thiol peroxidase (cbo0501 [tpx]),
rubredoxin
(cbo1252
[grxC]),
thioredoxin
reductase
(cbo1254
[trxB1]), a putative oxidative stress-related operon (cbo1330,
cbo1331, cbo1332, cbo1333), flavodoxin (cbo2778) and oxidoreduc-
tase (cbo3429), was observed. Furthermore, in the strongly-induced
cluster 3 (Fig. 3), genes putatively encoding a ferrous iron transport
mechanism (FeoAB) (cbo1028-cbo1031, cbo1076) were identified. Additionally, induction of another ferrous iron transport gene
(cbo1077 [feoA]) was observed in cluster 1 (Fig. 3), along with genes
encoding ferric uptake regulator (FUR) family proteins (cbo2560
and cbo3220), a ferritin (cbo1784 [ftnA]), an operon possibly related
to metal ion homeostasis (cbo2224-cbo2222, cbo2221), and two
ArsR-type regulators (cbo0182 and cbo0477). Finally, cbo1795
encoding the DNA-damage related LexA repressor was up-
regulated. The transcription of a large number of genes was markedly
affected 5 h after the temperature downshift. The results for these
genes are presented in Table S1, and elaborated in detail below in
the context of their predicted functional categories. Fatty acid metabolism. Of the 26 genes putatively related to
fatty acid metabolism, 9 were significantly up-regulated and 6
down-regulated (Fig. 4). The strongest induction of all ORFs
represented in the microarray was observed for genes within the
putative operon cbo2592-cbo2581. The first gene therein, cbo2592,
was 270-fold induced. Functional predictions for several of the
operon genes suggest the operon to be involved in fatty acid
biosynthesis; however, the rest of the genes within the cluster had
poor resemblance to characterized fatty acid metabolism-related
mechanisms. Therefore, the definite function of this cluster
remains to be confirmed. All six down-regulated genes in this
functional category represented the gene cluster cbo3605-cbo3594. Most genes within this cluster encode proteins that resemble
components of well-characterized bacterial fatty acid synthesis
pathways. Thus, the down-regulated cbo3605-cbo3594 cluster Genes related to processes of vegetative growth. Cluster
4 (Fig. 3) included strongly-repressed genes mostly involved in
vegetative
growth. Genes putatively
involved
in nucleotide Genes related to processes of vegetative growth. Cluster
4 (Fig. 3) included strongly-repressed genes mostly involved in
vegetative
growth. Genes putatively
involved
in nucleotide February 2014 | Volume 9 | Issue 2 | e89958 PLOS ONE | www.plosone.org 3 Transcriptome of C. botulinum in Cold Shock Figure 2. Confirmation of DNA microarray results with
quantitative real-time reverse-transcription PCR (RT-qPCR). The Effects of Prolonged Cold Exposure on the
Transcriptome of C. botulinum ATCC 3502 doi:10.1371/journal.pone.0089958.g002 Another strongly up-regulated gene (180-fold) 5 h after the cold
shock was cbo0389 putatively encoding an amino acid permease. Additionally, the transcription of another putatively amino acid
permease-encoding cbo0343 was 4.5-fold induced. Regulatory functions. In total, 30 genes with predicted
regulatory roles were identified as differentially transcribed, 22 of
which were up- and 8 down-regulated (Fig. 4). Of these, genes with
obvious associations with specific functional categories mentioned
above are presented under their respective sections. synthesis and metabolism (cbo1808 [cmk], cbo1864 [deoD], cbo2072
[codA], cbo2073 [codB], cbo2821 [pyrR], cbo3081 [hpt], cbo3241-
cbo3238
[pyrBICF],
cbo3296
[guaB],
cbo3544
[prsA]),
energy
metabolism (cbo1489, cbo1498, cbo3242, cbo3244, cbo3288 [acdA]),
transcription or translation (cbo2434 [tsf], cbo2939 [dnaG]), cobal-
amin biosynthesis (cbo0410, cbo0914-cbo0916 [cobQ-cbiB-cobD]), or
cell wall synthesis (cbo0791 [dapB], cbo1827, cbo3100) were
included. In addition, genes putatively co-transcribed with the
strongly-affected genes in cluster 4, were identified in the
moderately-repressed cluster 2. These included cbo1499-cbo1500
(metN2-metI2), cbo2938 (sigA), cbo3237-cbo3235 (pyrKDE), cbo3243,
cbo3287-cbo3286 (etfB2A2), cbo3295 (guaA), as well as the cobalt
transporters (cbo0917-cbo0919 [cbiMNQ], cbo3450-cbo3449) and all
the other genes putatively related to cobalamin metabolism
(cbo1492-cbo1494 [cobW-cbo1493-mtbC] and cbo1495 [mtbB]). synthesis and metabolism (cbo1808 [cmk], cbo1864 [deoD], cbo2072
[codA], cbo2073 [codB], cbo2821 [pyrR], cbo3081 [hpt], cbo3241-
cbo3238
[pyrBICF],
cbo3296
[guaB],
cbo3544
[prsA]),
energy
metabolism (cbo1489, cbo1498, cbo3242, cbo3244, cbo3288 [acdA]),
transcription or translation (cbo2434 [tsf], cbo2939 [dnaG]), cobal-
amin biosynthesis (cbo0410, cbo0914-cbo0916 [cobQ-cbiB-cobD]), or
cell wall synthesis (cbo0791 [dapB], cbo1827, cbo3100) were
included. In addition, genes putatively co-transcribed with the
strongly-affected genes in cluster 4, were identified in the
moderately-repressed cluster 2. These included cbo1499-cbo1500
(metN2-metI2), cbo2938 (sigA), cbo3237-cbo3235 (pyrKDE), cbo3243,
cbo3287-cbo3286 (etfB2A2), cbo3295 (guaA), as well as the cobalt
transporters (cbo0917-cbo0919 [cbiMNQ], cbo3450-cbo3449) and all
the other genes putatively related to cobalamin metabolism
(cbo1492-cbo1494 [cobW-cbo1493-mtbC] and cbo1495 [mtbB]). Several key regulators of central metabolism were affected by
temperature downshift. A 7.8-fold induction of the GTP-sensing
transcriptional pleiotropic repressor codY (cbo2436) was observed. Additionally, significant down-regulation was observed for relA
(cbo3059) encoding a GTP pyrophosphokinase that has a function
in triggering de-repression of CodY. Among the cold-induced regulatory genes with no obvious
association with structural or functional loci was cbo0097 predicted
to encode a PadR-type DNA-binding transcriptional regulator. The Effects of Prolonged Cold Exposure on the
Transcriptome of C. botulinum ATCC 3502 Log2 fold changes of transcript levels measured with DNA microarrays
(x-axis) and RT-qPCR (y-axis) in C. botulinum ATCC 3502 cultures 1 h
after temperature downshift from 37uC to 15uC. 16S rrn transcript levels
were used as a normalization reference in the RT-qPCR. Linear
regression analysis showed an R2 correlation value of 0.93 between
the microarray and RT-qPCR transcription fold change results. doi:10.1371/journal.pone.0089958.g002 molecules) were also associated with the amino acid biosynthetic
pathways identified. The results for these genes are therefore also
presented here. In cluster 3 (Fig. 3), cbo2670-cbo2669 (argGH) encoding an
argininosuccinate
synthase
and
an
argininosuccinate
lyase,
respectively, which produce arginine (Arg) from L-citrulline, were
identified, with respective up-regulation of .46- and 39-fold. However, the genes encoding proteins involved in Arg catabolism,
i.e. arginine deiminases (cbo0065 [arcA1] and cbo1587 [arcA2]),
carbamate kinase and ornithine carbamoyltransferase (cbo2594-
cbo2593 [arcCB]), arginine/ornithine antiporter (cbo1588 [arcD]),
and Orn/Lys/Arg decarboxylase (cbo3116 [speA]) producing
agmatine from L-arginine, were significantly down-regulated. Additionally, the whole putative transcriptional unit cbo1882-1877
including the arginine repressor-encoding cbo1878 (argR) was
down-regulated. The glutamate dehydrogenase (cbo1811 [gluD]) converting L-
glutamate into 2-oxoglutarate and glutamine synthetase (cbo3563
[glnA]) synthesizing L-glutamine from L-glutamate were down-
regulated, whereas glutaminase (cbo2808 [glsA]) converting L-
glutamine to L-glutamate was up-regulated. Additionally, cbo3337-
cbo3336 (lysCA) and cbo1912-cbo1911 (dapF-cbo1911) contributing to
the production of L-lysine (Lys) from diaminopimelate were up-
regulated, whereas cbo1457-cbo1458 (murEF), which use diamino-
pimelate for peptidoglycan biosynthesis, were down-regulated. Furthermore, differential transcript levels for several putative D-
proline reductase-related operons were observed. Of these,
cbo2482-cbo2479
(prdC2-cbo2481-prdA2-cbo2479)
and
cbo2466-
cbo2462 (prdC1-cbo2465-prdA1-cbo2463-prdB2) both encoding elec-
tron transfer subunits of proline reductase, subunits of proline
reductase, and hypothetical proteins, cbo2461 (prdB2) encoding a
subunit of proline reductase, and cbo2476-cbo2474 (prdDEF)
encoding D-proline reductase and proline racemase were up-
regulated. In contrast, cbo2489-cbo2488 (prdC4-prdA4) and cbo2484-
cbo2483 (prdC3-prdA3) encoding an electron transfer subunit of
proline reductase and a partial proline reductase were both down-
regulated Figure 2. Confirmation of DNA microarray results with
quantitative real-time reverse-transcription PCR (RT-qPCR). Log2 fold changes of transcript levels measured with DNA microarrays
(x-axis) and RT-qPCR (y-axis) in C. botulinum ATCC 3502 cultures 1 h
after temperature downshift from 37uC to 15uC. 16S rrn transcript levels
were used as a normalization reference in the RT-qPCR. Linear
regression analysis showed an R2 correlation value of 0.93 between
the microarray and RT-qPCR transcription fold change results. The Effects of Prolonged Cold Exposure on the
Transcriptome of C. botulinum ATCC 3502 This gene, showing a 2 and 10-fold induction 1 h and 5 h after
cold shock, respectively, is located within a cluster of three genes,
including cbo0096 encoding a putative uncharacterized membrane
protein and cbo0098 encoding a putative zinc-dependent hydro-
lase. Similar induction pattern was observed for cbo0096 and
cbo0097 but not cbo0098, suggesting bicistronic transcription of
cbo0097-cbo0096. Amino acid metabolism. Ten genes encoding amino acid
biosynthesis proteins were differentially transcribed 5 h after the
cold shock (Fig. 4). However, based on pathway predictions in the
KEGG database (http://www.genome.jp/kegg/), a significant
number of differentially transcribed genes from other functional
classes (e.g. Transport/binding proteins or Degradation of small The cbo0477 predicted to encode an ArsR-type transcriptional
regulator appears to be co-transcribed with cbo0478 encoding a February 2014 | Volume 9 | Issue 2 | e89958 PLOS ONE | www.plosone.org February 2014 | Volume 9 | Issue 2 | e89958 4 Transcriptome of C. botulinum in Cold Shock Figure 3. K-means clusters of up- or down-regulated genes 1 h and 5 h after temperature downshift in C. botulinum ATCC 3502
means clustering of significantly up- or down-regulated genes 1 h and 5 h after temperature downshift from 37uC to 15uC in C. botulinum ATCC 3
into four clusters. The sub-clusters were further subjected to hierarchical clustering. doi:10.1371/journal.pone.0089958.g003 Figure 3. K-means clusters of up- or down-regulated genes 1 h and 5 h after temperature downshift in C. botulinum ATCC 350
means clustering of significantly up- or down-regulated genes 1 h and 5 h after temperature downshift from 37uC to 15uC in C. botulinum ATCC
into four clusters. The sub-clusters were further subjected to hierarchical clustering. doi:10.1371/journal.pone.0089958.g003 Figure 3. K-means clusters of up- or down-regulated genes 1 h and 5 h after temperature downshift in C. botulinum ATCC 3502. K-
means clustering of significantly up- or down-regulated genes 1 h and 5 h after temperature downshift from 37uC to 15uC in C. botulinum ATCC 3502
into four clusters. The sub-clusters were further subjected to hierarchical clustering. doi:10.1371/journal.pone.0089958.g003 February 2014 | Volume 9 | Issue 2 | e89958 PLOS ONE | www.plosone.org 5 Transcriptome of C. botulinum in Cold Shock Figure 4. Distribution of significantly up- or down-regulated genes in functional categories. The number of si
regulated and unaffected genes 5 h after temperature downshift from 37uC to 15uC in C. The Effects of Prolonged Cold Exposure on the
Transcriptome of C. botulinum ATCC 3502 botulinum ATCC 3502 divided into f
bar lengths portray the percentage of affected and unaffected genes of all genes assigned to each functional category. T
were as described in the original annotation of the ATCC 3502 genome [12]. Genes with no expression data available due to
are assigned to the ‘‘Not affected/unknown’’ category. doi:10.1371/journal.pone.0089958.g004 Figure 4. Distribution of significantly up- or down-regulated genes in functional categories. The number of significantly up- or down-
regulated and unaffected genes 5 h after temperature downshift from 37uC to 15uC in C. botulinum ATCC 3502 divided into functional categories. The
bar lengths portray the percentage of affected and unaffected genes of all genes assigned to each functional category. The functional categories
were as described in the original annotation of the ATCC 3502 genome [12]. Genes with no expression data available due to poor array hybridization
are assigned to the ‘‘Not affected/unknown’’ category. doi:10.1371/journal.pone.0089958.g004 Figure 4. Distribution of significantly up- or down-regulated genes in functional categories. The number of significantly up- or down-
regulated and unaffected genes 5 h after temperature downshift from 37uC to 15uC in C. botulinum ATCC 3502 divided into functional categories. The
bar lengths portray the percentage of affected and unaffected genes of all genes assigned to each functional category. The functional categories
were as described in the original annotation of the ATCC 3502 genome [12]. Genes with no expression data available due to poor array hybridization
are assigned to the ‘‘Not affected/unknown’’ category. d i 10 1371/j
l
0089958 004 Figure 4. Distribution of significantly up- or down-regulated genes in functional categories. The number of significantly up- or down-
regulated and unaffected genes 5 h after temperature downshift from 37uC to 15uC in C. botulinum ATCC 3502 divided into functional categories. The
bar lengths portray the percentage of affected and unaffected genes of all genes assigned to each functional category. The functional categories
were as described in the original annotation of the ATCC 3502 genome [12]. Genes with no expression data available due to poor array hybridization
are assigned to the ‘‘Not affected/unknown’’ category. doi:10.1371/journal.pone.0089958.g004 suggest that the regulatory proteins CBO0477 and CBO0558A
have a role in cold tolerance of C. botulinum ATCC 3502. suggest that the regulatory proteins CBO0477 and CBO0558A
have a role in cold tolerance of C. botulinum ATCC 3502. putative heavy metal-translocating ATPase. Discussion Transcriptomic analysis of the cold-shock response of C. botulinum ATCC 3502 revealed induction of metabolic pathways
with established cold-related functions, as well as the induction of a
large number of uncharacterized genes. Induction of 16 genes was
observed already 1 h after the cold shock, indicating a specialized
acute response to temperature downshift. All of these 16 genes
showed sustained and further increased induction upon extended
cold exposure. The data suggest an important role for their
encoded mechanisms not only in the rapid cold-shock response,
but also in long-term cold adaptation. At the later stage of cold
adaptation, marked effects on the transcriptome were observed,
suggesting extensive metabolic remodeling upon cold shock. The Effects of Prolonged Cold Exposure on the
Transcriptome of C. botulinum ATCC 3502 A 2.6-fold induction
1 h after the cold shock was observed for cbo0477, the transcript
levels further increasing to 30-fold 5 h after the cold shock. One of the most strongly induced genes 1 h after the cold shock
was cbo0558A. Its induction at 1 h was 8.5-fold in the microarray
experiment, but was shown to be as high as 36-fold in RT-qPCR
analysis (Fig. 2). The transcript levels of cbo0558A further increased
to 35-fold in the microarray data 5 h after the cold shock, which
represented the highest level of induction among the regulator-
encoding genes. Similar transcriptional patterns were observed for
cbo0558A, cbo0559 and cbo0560, suggesting a transcriptional link
between these genes. February 2014 | Volume 9 | Issue 2 | e89958 Inactivation of the Cold-induced Putative Transcriptional
Regulators cbo0477 and cbo0558A Results in Deteriorated
Cold Tolerance botulinum ATCC 3502 wild type, and cbo0097, cbo0477 and cbo0558A mutants at 17uC. Growth of C. botulinum
ATCC 3502 wild type (WT, A-C), cbo0097 mutant (A), cbo0477 mutant (B), and cbo0558A mutant (C) cultures in TPGY broth at 17uC. Antisense (AS) and
sense (S) orientation mutants were constructed for each gene. The error bars represent the minimum and maximum OD600 values measured for three
independent biological replicates. doi:10.1371/journal.pone.0089958.g005 of increased mRNA stability that hampers translation [16,17]. Roles in ribosome biogenesis have also been proposed [18]. No
reports on the role of DEAD-box helicases in cold tolerance of C. botulinum are available. However, a strong induction of DEAD-box
helicase genes upon cold stress has been observed at least in
Bacillus cereus [19], B. subtilis [7], Listeria monocytogenes [20] and
Yersinia pseudotuberculosis [21]. These studies used also mutational
analysis to support the importance of DEAD-box helicases in cold
tolerance of these organisms. The rapid and strong induction of
deaD upon cold shock suggests a role in cold tolerance in C. botulinum;
this
hypothesis
is
currently
being
tested
in
our
laboratory. Among the mechanisms readily induced 1 h after cold shock
was the uptake of compatible solutes. The transcript levels of these
genes further increased 5 h after the cold shock. In addition,
cbo1131 (opuC/opuD) putatively encoding another choline/carni-
tine/betaine transporter, was significantly up-regulated 5 h after
the cold shock. Induction of the high-affinity transport mecha-
nisms for compatible solutes, such as glycine betaine, carnitine,
choline and proline, has been linked to osmotic and cold stress in
L. monocytogenes [14–15] and B. subtilis [23] and explained by cryo-
and osmoprotective functions of these compounds. Cold-induction
of related genes in C. botulinum ATCC 3502 suggests utilization of
compatible solutes as a means to counter cold stress. y
C. botulinum ATCC 3502 possesses three cold-shock protein
(Csp) homologues (cspABC) of which cspB was suggested to encode
the major cold-related Csp [8]. Csps have roles in destabilizing
cold-induced mRNA secondary structures thereby facilitating
efficient translation [5]. Csps are found in the majority of
organisms; however, interestingly, they seem to be absent from
(Group II) type E C. botulinum genomes [22]. Significant induction
of all three Csp-encoding genes already 1 to 30 min after cold
shock was observed by So¨derholm et al. [8]. Similarly, induction of
both cspA and cspB was observed in the current study. Inactivation of the Cold-induced Putative Transcriptional
Regulators cbo0477 and cbo0558A Results in Deteriorated
Cold Tolerance To test whether the strongly up-regulated genes encoding DNA-
binding regulators have a role in cold tolerance of C. botulinum
ATCC 3502, we inactivated cbo0097, cbo0477 and cbo0558A and
compared the growth of such mutants and the wild-type strain at
17uC for 8 days. Inactivation of cbo0097 did not result in
significantly decreased cold sensitivity: The final OD600 of the
antisense mutant was somewhat higher than that of the sense
mutant and the wild-type strain, whereas the sense mutant showed
a similar growth pattern to the wild-type strain (Fig. 5A). In
contrast, impaired growth at 17uC was observed for all the cbo0477
(Fig. 5B) and cbo0558A (Fig. 5C) mutants. No difference in growth
at 37uC was observed between the wild-type strain and any of the
mutants (data not shown). Insertional inactivation was performed
in two independent sites and orientations to confirm the mutation
as the primary cause behind the observed phenotype. The results Analysis of the differentially transcribed genes by k-means
clustering
suggested
biological
relevance
especially
for
the
strongly-affected clusters 3 and 4. Cluster 3 included up-regulated
genes related to previously identified bacterial cold tolerance
mechanisms, e.g. compatible solute transport [14,15], cold shock
proteins [5,8], and a DEAD-box RNA helicase [16], verifying the
relevance of the results. Moreover, cluster 4 included down-
regulated genes putatively involved in central metabolic and
vegetative growth-related processes, such as nucleotide biosynthe-
sis and inter-conversions, consistent with a shutdown of central
metabolism associated with cold-shock induced growth arrest and
adaptation phase [5]. The DEAD-box proteins have been shown to have a significant
role in cold tolerance in a multitude of organisms. They function
as RNA helicases and chaperones and thereby counter the effects February 2014 | Volume 9 | Issue 2 | e89958 February 2014 | Volume 9 | Issue 2 | e89958 6 PLOS ONE | www.plosone.org Transcriptome of C. botulinum in Cold Shock Figure 5. Growth of C. botulinum ATCC 3502 wild type, and cbo0097, cbo0477 and cbo0558A mutants at 17uC. Growth of C. botulinum
ATCC 3502 wild type (WT, A-C), cbo0097 mutant (A), cbo0477 mutant (B), and cbo0558A mutant (C) cultures in TPGY broth at 17uC. Antisense (AS) and
sense (S) orientation mutants were constructed for each gene. The error bars represent the minimum and maximum OD600 values measured for three
independent biological replicates. doi:10.1371/journal.pone.0089958.g005 Figure 5. Growth of C. February 2014 | Volume 9 | Issue 2 | e89958 Inactivation of the Cold-induced Putative Transcriptional
Regulators cbo0477 and cbo0558A Results in Deteriorated
Cold Tolerance Down-
regulation of the primary exponential-phase sigma factor SigA
(cbo2938) is consistent with growth arrest-related shutdown of
central metabolism. Almost an equal number of down- and up-
regulated genes in the transport/binding proteins, fatty acid
biosynthesis and cell envelope classes (Fig. 4) can be explained by
changes in cell processes due to the slower growth rate. Among the
down-regulated transport/binding protein-encoding genes, ones
involved in carbohydrate, cobalt or uracil uptake were identified,
i.e. those needed for active growth. However, genes putatively
encoding certain amino acid transporting mechanisms were up-
regulated. The possible roles of these up-regulated mechanisms in
bacterial cold tolerance are discussed below. Several genes with functions predicted in oxidative stress
response were significantly cold-induced. Similarly, oxidative
stress response was also induced in B. subtilis upon cold shock
[6] and in L. monocytogenes and Psychrobacter arcticus during growth at
low temperature [36,37]. Recently, the presence of secondary
oxidative stress was demonstrated in B. subtilis exposed to a variety
of environmental stress conditions [38,39]. Our observation on the
induction of the oxidative stress response upon cold shock strongly
supports the hypothesis of secondary oxidative stress as a central
player in ‘‘general’’ stress. Therefore, we suggest an important role
for induction of the oxidative stress response in countering cold
stress in C. botulinum. Down-regulation of most cobalamin (coenzyme B12) synthesis
genes and the closely-related cobalt uptake mechanisms was
observed. The main function of cobalamin in anaerobic bacteria is
thought
to
be
anaerobic
fermentation
of
small
molecules
(ethanolamine, propanediol or glycerol) and simultaneous gener-
ation of an electron sink subsequently balancing redox reactions
[45]. In C. acetobutylicum, the cobalt- and cobalamin-related genes
were observed to be down-regulated upon entry into stationary
phase [46]. The down-regulation of cobalamin biosynthesis genes
in our experiment can be explained by cessation of growth, or by
changes in redox balance-related reactions due to temperature
downshift as discussed above. In addition to mechanisms directly related to oxidative stress,
genes related to iron transport and storage were induced upon
cold shock, along with two FUR family regulator genes. Induction
of genes for ferrous iron transporters and iron-storing ferritins,
similar to the gene set induced upon cold stress in our experiment,
was observed upon O2 stress in Clostridium acetobutylicum [40]. Iron
possesses a conflictive role upon oxidative stress: ferrous iron feeds
the Fenton reaction, resulting in production of toxic hydroxyl
radicals [41]. Inactivation of the Cold-induced Putative Transcriptional
Regulators cbo0477 and cbo0558A Results in Deteriorated
Cold Tolerance Upon DNA damage,
RecA binds to the resulting single-stranded DNA molecule and
facilitates the inactivation of the transcriptional repressor LexA
[43]. As a consequence, the LexA-mediated repression of SOS-
regulated loci is liberated and the SOS response is activated [43]. Our data suggest activation of the SOS response upon cold shock,
implying that direct or indirect DNA damage might result from
cold stress in C. botulinum. Direct DNA damage can arise from
nucleophilic attacks by hydroxyl radicals. As discussed above, the
observed induction of oxidative stress-related genes supports the
hypothesis of secondary oxidative stress as a player in cold stress. The cold-induction of the SOS response further supports this
hypothesis. Considering food-borne pathogens, the increased
cross-protective robustness arising from sub-lethal stress due to
control measures applied in minimal food processing is of concern. This phenomenon was recently shown in Bacillus weihenstephanensis,
where cultures grown at 7uC showed increased tolerance towards
severe oxidative stress, compared to cells cultured at 30uC [44]. Thus, future research on the role of oxidative stress as a player in
the general stress response and in cold-shock response of C. botulinum is of special interest. In addition to cbo2585, two other FabH homologues (cbo0718,
cbo3604) were identified in the C. botulinum ATCC 3502 genome. They both exhibited down-regulation 5 h after the temperature
downshift. The cbo3604 resides within a cluster of genes with
predicted functions in FA synthesis, suggesting cbo3604 may
encode the primary FabH enzyme of C. botulinum ATCC 3502. Remarkably, this cluster was significantly down-regulated upon
cold shock. This observation, together with the extremely strong
cold-induction of cbo2585 and the adjoining gene cluster, could
suggest cbo2592-cbo2581 encodes an alternative cold-triggered FA
biosynthesis pathway. Thus, in contrast to the temperature-
dependent change in FabH primer preference utilized by L. monocytogenes [30], C. botulinum could counter cold-induced lipid
solidification by switching to a totally different cold-activated FA
biosynthesis machinery. Unfortunately, to the authors’ knowledge,
the function and substrate-specificity of clostridial FabH enzymes
has not been characterized. Furthermore, reports on the utilization
of BCAA and the presence of BCFA in C. botulinum and in the
closely-related Clostridium sporogenes are somewhat contradicting
[32–35]. Thus, further characterization of the FA biosynthesis
mechanisms and their role in cold tolerance in C. botulinum is
warranted. Upon cold shock, growth is halted until bacteria have adjusted
to the changed temperature, with several central metabolic
processes being shut down for the adaptation period [5]. Inactivation of the Cold-induced Putative Transcriptional
Regulators cbo0477 and cbo0558A Results in Deteriorated
Cold Tolerance Induction of
cspB was relatively mild (2.2-fold) 1 h after temperature downshift,
however, the transcript levels increased to suggest 4.7-fold up-
regulation 5 h after the cold shock. No significant change in
transcript levels of cspC was observed 1 to 5 h after cold shock in
the current study, which is in agreement with the findings by
So¨derholm et al. [8]. A considerable induction of cbo2592-cbo2581, putatively encod-
ing fatty acid (FA) metabolism-related proteins, was observed upon
cold shock. The cbo2585 encoding a homologue to FabH (3-
oxoacyl-[acyl-carrier-protein] synthase 3), responsible for initiation
of FA synthesis, is present in this cluster together with two putative
acyl-carrier protein genes (cbo2583 and cbo2589). While the genetic
arrangement suggests this cluster to be involved in FA metabolism,
many of the genes within the cluster have poor homology to
previously characterized FA metabolism mechanisms. Moreover,
none of the cluster components have been allocated to any specific
established pathway in the KEGG database. Thus, the ultimate
function of this operon remains to be characterized. An important negative effect of temperature downshift in the
bacterial cell is the solidification of the lipid cell membrane [24]. Bacteria have developed several strategies to restore membrane
fluidity
and
subsequent
biological
function. These
include February 2014 | Volume 9 | Issue 2 | e89958 PLOS ONE | www.plosone.org February 2014 | Volume 9 | Issue 2 | e89958 7 Transcriptome of C. botulinum in Cold Shock uptake and storage mechanisms and the FUR family regulators in
cold tolerance of C. botulinum remain to be elucidated. desaturation of existing membrane FAs by an oxygen-dependent
lipid desaturase system [25–27], increasing the de novo synthesis of
unsaturated and branched FAs, and by shortening the acyl chain
length of the newly-synthesized FAs [24,28]. In the Gram-positive
L. monocytogenes and B. subtilis, branched-chain fatty acids (BCFA)
are synthesized from branched-chain acyl-coenzyme A (CoA)
primers, which are derived from the branched-chain amino acids
(BCAA) valine, leucine and isoleucine by branched-chain a-keto
acid dehydrogenase (Bkd) enzyme complex [24,29–31]. The FabH
of L. monocytogenes shows increased affinity to BCAA-CoA primers
upon temperature downshift [30], resulting in increased mem-
brane BCFA content and subsequent increase in membrane
fluidity. Several genes reported to be repressed by the SOS transcrip-
tional repressor LexA in B. subtilis [42] were cold-induced in C. botulinum. These included lexA, cbo2405 (recA), and cbo2818 (dinB,
putatively encoding DNA polymerase IV). February 2014 | Volume 9 | Issue 2 | e89958 Transcriptome of C. botulinum in Cold Shock The induction of several previously uncharacterized DNA-
binding regulatory protein-encoding genes upon cold shock
suggested the presence of as yet unidentified cold tolerance
mechanisms for C. botulinum. Therefore, we further analyzed three
rapidly and/or strongly-induced genes representing structurally
distinct regulator sub-families. Insertional inactivation of cbo0477
resulted in increased sensitivity to low temperature, as demon-
strated by growth curve analysis (Fig. 5B). The genomic context of
cbo0477 suggests this gene to have a role in metal ion homeostasis,
thus having possible implications on cold tolerance as discussed
above
in
context
of
iron
uptake
and
storage. Moreover,
inactivation of the highly-induced cbo0558A expectedly resulted
in deteriorated growth at 17uC (Fig. 5C), while growth in optimal
conditions was not affected. The strong and sustained induction of
cbo0558A combined with the cold-sensitive phenotypes of mutants
of this gene, suggests an important function for the CBO0558A
regulator in cold tolerance of C. botulinum, and warrants further
investigation. A homologue for cbo558A is found across Group I C. botulinum genomes within an operon of three genes (homologous to
cbo0558A, cbo0559 and cbo0560), the other two of them putatively
encoding components of an uncharacterized ABC transporter
system. The induction pattern of cbo0558A together with its
apparent conservation presents it as an interesting candidate for a
biomarker for cold-induced robustness in C. botulinum. Whil
t bl
ld i d
ti
f
b 0097
b
d
it Strongly-induced transcription of two putative amino acid
permeases (cbo0343 and cbo0389) was observed. Amino acid
permeases transport (specific) amino acids into the cell. However,
neither of the two proteins had significant homology to any
characterized proteins of related organisms, thus, the specificity of
these transporters remains unknown. However, the observed
remodeling of the amino acid metabolism to conserve Arg, Glu,
Lys, and Pro suggests that the transporters encoded by cbo0343
and cbo0389 could transport these amino acids. Low temperature induced the transcription of codY in C. botulinum
ATCC 3502. CodY, the global stationary
phase
regulator, is a key player in the central metabolic processes in
several low-GC Gram-positive bacteria [53]. CodY regulates the
transcription of over a hundred genes, including itself [54–57],
generally repressing them during rapid growth and de-repressing
them under nutritionally poor conditions (stringent response). Depending on the organism, the CodY repressor activity is
enhanced by GTP and/or BCAAs. Transcriptome of C. botulinum in Cold Shock cbo0803-cbo0801 encoding the hemagglutinins was observed, and
no significant changes were observed for cbo0804 (botR), cbo0805
(ntnh), or cbo0806 (botA). possibly also low, resembling stringent conditions. Indeed, L. monocytogenes was suggested to suffer from amino acid starvation
(i.e. stringent conditions) upon cold stress [36]. Thus, a temper-
ature downshift can be hypothesized to possess elements from both
nutrient-rich and stringent conditions, resulting in partial de-
repression of the CodY regulon. As observed for arginine, the transcriptional changes of genes
involved in glutamic acid (Glu) metabolism appeared to aim at
increasing the amount of Glu within the cell. Furthermore, similar
behavior was observed for lysine-associated genes, suggesting Lys
accumulation upon cold stress. Finally, the pathway for Stickland-
type reduction [49] of D-proline was up-regulated. CodY-mediated control of virulence has been demonstrated in
different organisms [56,62–64]. However, as discussed above, no
evident changes of transcription were observed for genes involved
in virulence upon cold shock in C. botulinum ATCC 3502. In L. monocytogenes, a similar induction of codY at low temperature has
been observed without de-repression of certain CodY-regulated
genes [50]. These data further support the partial activation of the
CodY regulon upon cold shock, suggesting significant plasticity for
CodY-mediated regulation in C. botulinum under different envi-
ronmental conditions. In contrast to our findings, studies on B. subtilis [6] and L. monocytogenes [50] revealed increased expression of BCAA biosyn-
thesis genes at low temperature, without any further cold-triggered
effects on amino acid metabolism. In E. coli, decreased expression
of Arg biosynthesis and degradation genes, as well as of Glu
synthase genes was observed at low temperature [5]. Our data
suggest that unlike these organisms, C. botulinum strives to
incorporate Arg, Glu and Lys within the cell after temperature
downshift. Bacteria use Arg, Glu and/or Lys decarboxylation to
cope with acid stress [51]. Thus, a possible goal for Arg, Glu, and
Lys conservation at low temperature could be their use as
compatible solutes in C. botulinum ATCC 3502. The induction of
the D-proline reduction pathway could arise from the probable use
of this reaction as an electron sink, thus playing a role in
controlling the redox balance of the cell [52]. Induction of the D-
proline reductase is thus in agreement with induction of the other
redox balance-related genes discussed above. Transcriptome of C. botulinum in Cold Shock In stringent conditions, the
higher amount of uncharged tRNAs activates RelA-mediated
conversion of GTP to guanosine 59-triphosphate 39-diphosphate
or guanosine 59-diphosphate 39-diphosphate (pppGpp and ppGpp,
hereafter abbreviated as [p]ppGpp) subsequently lowering the
GTP level within the cell and triggering de-repression of CodY-
regulated genes [53,58,59]. The induction of several previously uncharacterized DNA-
binding regulatory protein-encoding genes upon cold shock
suggested the presence of as yet unidentified cold tolerance
mechanisms for C. botulinum. Therefore, we further analyzed three
rapidly and/or strongly-induced genes representing structurally
distinct regulator sub-families. Insertional inactivation of cbo0477
resulted in increased sensitivity to low temperature, as demon-
strated by growth curve analysis (Fig. 5B). The genomic context of
cbo0477 suggests this gene to have a role in metal ion homeostasis,
thus having possible implications on cold tolerance as discussed
above
in
context
of
iron
uptake
and
storage. Moreover,
inactivation of the highly-induced cbo0558A expectedly resulted
in deteriorated growth at 17uC (Fig. 5C), while growth in optimal
conditions was not affected. The strong and sustained induction of
cbo0558A combined with the cold-sensitive phenotypes of mutants
of this gene, suggests an important function for the CBO0558A
regulator in cold tolerance of C. botulinum, and warrants further
investigation. A homologue for cbo558A is found across Group I C. botulinum genomes within an operon of three genes (homologous to
cbo0558A, cbo0559 and cbo0560), the other two of them putatively
encoding components of an uncharacterized ABC transporter
system. The induction pattern of cbo0558A together with its
apparent conservation presents it as an interesting candidate for a
biomarker for cold-induced robustness in C. botulinum. At low temperature, the translational capacity of the cell is
hampered due to decreased tRNA synthesis, thus lowering the
ratio of uncharged to charged tRNAs, a situation analogous to
nutrient-rich conditions. Indeed, several aminoacyl-tRNA genes
(cbo2976 [selA], cbo3503 [proS2], cbo3054 [aspS], cbo3055 [hisS],
cbo3265 [gatB]) were found to be down-regulated. Furthermore, a
decrease in (p)ppGpp levels, with additional effects on the
expression of cold shock protein-encoding genes at low temper-
ature, has been reported [60,61]. Thus, low temperature can
partially
resemble
nutrient-rich
conditions
in
the
form
of
decreased uncharged-to-charged tRNA ratio. This subsequently
could decrease RelA activity and result in lower (p)ppGpp levels. However, at low temperature, due to the slower growth rate and
down-regulation of purine metabolism genes, the GTP level is While notable cold-induction of cbo0097 was observed, its
inactivation did not result in cold-sensitivity. Inactivation of the Cold-induced Putative Transcriptional
Regulators cbo0477 and cbo0558A Results in Deteriorated
Cold Tolerance On the other hand, detoxifying and repair enzymes
commonly use iron as a cofactor, thus presenting an increased
need for iron upon (oxidative) stress [41]. Hillmann et al. [40]
suggest an induction of iron uptake mechanisms at O2 stress in C. acetobutylicum to be a response to increased iron demand arising
from activation of detoxifying and repair enzymes. A similar goal
can thus be hypothesized for the cold-induction of iron uptake in
C. botulinum. However, the exact role and function of induced iron Extensive remodeling of amino acid uptake and utilization
patterns was observed upon temperature downshift. As the
catabolism and biosynthesis of Arg are activated and repressed,
respectively, by the down-regulated ArgR, the remodeling of the
Arg metabolism upon temperature downshift by C. botulinum seems
to aim in Arg conservation rather than degradation. Arg is an
essential amino acid for Group I C. botulinum [47], and a high Arg
level in the culture medium has been proposed to suppress
neurotoxin production and protease activity [48]. However, the
apparent shift towards Arg accumulation did not result in down-
regulation of the neurotoxin cluster genes: slight up-regulation of February 2014 | Volume 9 | Issue 2 | e89958 February 2014 | Volume 9 | Issue 2 | e89958 PLOS ONE | www.plosone.org 8 February 2014 | Volume 9 | Issue 2 | e89958 Transcriptome of C. botulinum in Cold Shock We have previously identified cold-induction of genes encoding
the two-component systems (TCSs) CBO0366/CBO0365 and
CBO2306/CB02307, and the alternative sigma factor SigK
(cbo2541) [9–11]. Further characterization showed these mecha-
nisms to have a role in cold tolerance of C. botulinum [9–11]. Significant induction of the cbo0364-cbo0366 operon was also
observed in the present DNA microarray experiment, albeit the 5-
h log2 fold changes fell slightly outside of the defined cut-off value
of 2. However, of all putative TCSs of C. botulinum ATCC 3502,
this TCS showed the strongest induction upon temperature
downshift. A modest induction was also observed for cbo2306,
while no significant induction was observed for cbo2307 or
cbo2541. In our previous RT-qPCR studies, however, the
transcription of cbo2307 or cbo2541 reached a 3.0 to 4.4-fold
induction 5 h after cold shock [10,11]. The RT-qPCR verification
of the DNA microarray data suggests that differences in transcript
levels are more modestly interpreted by the microarray experi-
ment than by RT-qPCR. The apparent discrepancy can be
attributed to the distinct normalization procedures. However, a
good correlation between the microarray and RT-qPCR data was
observed and suggests that the main findings discussed are reliable. We have previously identified cold-induction of genes encoding
the two-component systems (TCSs) CBO0366/CBO0365 and
CBO2306/CB02307, and the alternative sigma factor SigK
(cbo2541) [9–11]. Further characterization showed these mecha-
nisms to have a role in cold tolerance of C. botulinum [9–11]. Significant induction of the cbo0364-cbo0366 operon was also
observed in the present DNA microarray experiment, albeit the 5-
h log2 fold changes fell slightly outside of the defined cut-off value
of 2. However, of all putative TCSs of C. botulinum ATCC 3502,
this TCS showed the strongest induction upon temperature
downshift. A modest induction was also observed for cbo2306,
while no significant induction was observed for cbo2307 or
cbo2541. In our previous RT-qPCR studies, however, the
transcription of cbo2307 or cbo2541 reached a 3.0 to 4.4-fold
induction 5 h after cold shock [10,11]. The RT-qPCR verification
of the DNA microarray data suggests that differences in transcript
levels are more modestly interpreted by the microarray experi-
ment than by RT-qPCR. The apparent discrepancy can be
attributed to the distinct normalization procedures. However, a
good correlation between the microarray and RT-qPCR data was
observed and suggests that the main findings discussed are reliable. Transcriptomic Analysis with DNA Microarrays Array hybridizations for samples collected at mid-logarithmic
growth at 37uC and 1 h post-cold were performed in our previous
study [13], and samples collected 5 h post-cold shock were
similarly hybridized in this study. In brief, in situ -synthesized
Custom Gene Expression Microarrays (8x15K, Agilent Technol-
ogies) were designed to cover 3641 chromosomal (out of the total
of 3648) and all 19 plasmid-borne open reading frames (ORF) in
the ATCC 3502 genome [12]. Depending on the length of ORF, a
total of 3 to 14 60-mer oligonucleotide probes were designed for
each ORF. For array hybridizations, the three biological replicate samples
for each time point were labeled with either Cy3 or Cy5 as
described above; for each time point, two of the samples were
labeled with one dye and one with another. A total of 300 ng of
Cy3-labelled cDNA and 300 ng of Cy5-labelled cDNA were
mixed into a final volume of 18 ml, and 0.1 mg/ml salmon sperm
DNA (Life Technologies) was added into the reaction. The DNA
was denatured at 95uC for 2 min and chilled on ice, and finally
mixed with blocking agent (Hi-RPM GE Hybridization Kit,
Agilent Technologies) and hybridization buffer (Hi-RPM GE
Hybridization Kit, Agilent Technologies) as instructed by the
manufacturer. A volume of 50 ml of the mix was pipetted onto the
DNA microarrays. The arrays were hybridized at 65uC overnight,
and washed as instructed (Gene Expression Wash Buffer Kit,
Agilent Technologies). The cell pellets were thawed on ice for 5 min and used for RNA
extraction using the RNeasy Mini Kit or RNeasy Midi Kit
(Qiagen GmbH, Hilden, Germany) according to the manufactur-
er’s instructions. The cells were lysed with a solution containing
25 mg/ml lysozyme (Sigma-Aldrich, St. Louis, MO, USA) and
250 IU/ml mutanolysin (Sigma-Aldrich) in Tris-EDTA buffer
(pH 8.0, Fluka BioChemica, Buchs, Switzerland) and agitated at
37uC for 30 min. To ensure efficient removal of all genomic DNA,
an additional DNase treatment was carried out using the DNA-
free Kit (Life Technologies, Carlsbad, CA) according to manu-
facturer’s instructions. The RNA yield and purity (A260/A280) were checked using the
NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific
Inc., Waltham, MA). The A260/A280 ratio was .2.0 for all
samples. Integrity of RNA was confirmed with miniaturized gel
electrophoresis in the Agilent 2100 Bioanalyzer (Agilent Technol-
ogies Inc., Santa Clara, CA). The RNA integrity number was . 9.2 for all RNA samples. Conclusions Our results suggest a relatively small number of rapidly cold
shock-induced genes in C. botulinum ATCC 3502, with many
previously uncharacterized players. In contrast, the cold-triggered
transcriptional changes observed 5 h after the temperature
downshift were extensive, indicating a dramatic remodeling of
metabolism upon cold adaptation. While several mechanisms that
were cold-induced in C. botulinum have been previously identified
as cold-related also in other bacteria, identification of novel cold-
responsive regulators warrants further investigation. Importantly,
evidence suggesting a role for secondary oxidative stress and
associated protective responses in cold tolerance was obtained,
supporting the hypothesis of secondary oxidative stress as an
important part of a myriad of stress conditions. Identification of
genes with rapid and sustained induction upon cold stress could
aid in the discovery of biomarkers for detection of enhanced
tolerance and cross-protection against multiple stressors, and
should be of profound interest for food safety research. For RT-qPCR, a total of 500 ng of each RNA sample was used
for cDNA synthesis using the DyNAmo cDNA Synthesis Kit
(Thermo Fisher Scientific) as instructed by the manufacturer. Each
20-ml reaction, containing 15 ng/ml of random hexamers, 10 IU
of M-MuLV RNase H+ reverse transcriptase solution (Thermo
Fisher Scientific), and appropriate buffer containing dNTP and
MgCl2 in a final concentration of 5 mM (1 x, Thermo Fisher
Scientific), was incubated at 25uC for 10 min and 37uC for
30 min, then inactivated at 85uC for 5 min and finally chilled to
4uC. Two replicate RT reactions were made for each RNA
sample. The cDNAs were stored at 220uC until performing RT-
qPCR. cDNA Synthesis For DNA microarray analysis, a total of 2 mg of each RNA
sample was reverse-transcribed into cDNA and simultaneously
labeled with fluorescent dyes. In brief, each 30-ml labeling reaction
contained 0.2 mg/ml of random hexamers (Life Technologies),
0.01 M DTT (Life Technologies), 1.3 U/ml ribonuclease inhibitor
(Life Technologies), 0.5 mM dATP, dTTP and dGTP, 0.2 mM
dCTP, 1.7 nmol of Cy-3 or Cy-5 labeled dCTP (GE Healthcare,
Pittsburgh, PA), 13 U/ml of SuperScript III reverse transcriptase
(Life Technologies), and appropriate buffer (1 6 First Strand
Buffer, Life Technologies), and was incubated at 46uC for 3 h. RNA hydrolysis and reaction inactivation were performed by
addition of 0.5 mM EDTA and 10 ml of 0.1 M NaOH and
incubation at 70uC for 15 min. The reactions were subsequently
neutralized by addition of 10 ml of 0.1 M HCl. The cDNA was
purified with QIAquick PCR Purification Kit (Qiagen), with final
elution volume of 40 ml. The cDNA concentration of each sample
was measured with NanoDrop. Transcriptome of C. botulinum in Cold Shock Transcriptome of C. botulinum in Cold Shock Transcriptome of C. botulinum in Cold Shock essential for survival at low temperature, or they may be
compensated by other mechanisms. Experimental Procedures Cold Shock Culture Conditions and RNA Extraction
C. botulinum ATCC 3502 was used as a parent strain in this study
(Table 1). Cultures for RNA isolation for DNA microarray and
RT-qPCR analysis were prepared in our previous study [13]. Briefly, three independent wild-type C. botulinum ATCC 3502
cultures (biological replicates) were anaerobically grown in optimal
conditions
in
tryptone-peptone-glucose-yeast
extract
(TPGY)
broth until their optical density at 600 nm (OD600) reached
approximately 1. The cultures were subjected to temperature
downshift to 15uC and incubated anaerobically at 15uC for 5 h. Samples from each culture were anaerobically collected for total
RNA isolation immediately before the temperature downshift, and
1 h and 5 h after anaerobic incubation at 15uC. The samples were
collected into sterile plastic tubes containing 20% of the sample
volume of ice-cold ethanol-phenol (9:1) solution (Sigma-Aldrich,
St. Louis, MO), mixed thoroughly, and incubated on ice for
30 min. Cells were harvested by centrifugation (4uC, 80006g) for
5 min. The cell pellets were immediately frozen to 270uC until
RNA extraction. Transcriptome of C. botulinum in Cold Shock Therefore, the
mechanisms under the regulation of CBO0097 are probably not February 2014 | Volume 9 | Issue 2 | e89958 PLOS ONE | www.plosone.org 9 Transcriptomic Analysis with DNA Microarrays Finally, significantly differentially transcribed
ORFs with a log2 fold change ,22.0 or .2.0 at either time point
were considered to be included in the cold-regulated gene set. same transcript or functional category with the cold-regulated
ORFs when considering induction or repression of metabolic
pathways; these genes, however, were not included in the
clustering. The number of up- or down-regulated genes in each
functional category (Table S1) was determined. The category
assigned for each gene was derived from the original annotation of
the ATCC 3502 genome [12]. Transcriptomic Analysis with DNA Microarrays The slides were scanned (Axon GenePix Autoloader 4200 AL,
Molecular Devices LLC, Sunnyvale, CA) at 532 and 635 nm using
a 5-mm resolution. Image processing was done using the GenePix
Pro 6.0 software (Molecular Devices) and data analysis with R
limma package [65]. Foreground and local background intensities
of each spot were characterized by the mean and median pixel
values of the spot, respectively. Local background was subtracted PLOS ONE | www.plosone.org February 2014 | Volume 9 | Issue 2 | e89958 10 Transcriptome of C. botulinum in Cold Shock Table 1. Strains and plasmids used in this study. p
y
Strain or plasmid
Relevant properties
Source
Bacterial strains
C. botulinum ATCC 3502
Wild type parental strain
ATCC1
C. botulinum ATCC 3502 cbo0097::intron 124|125AS
Insertional disruption of cbo0097 at base 124 in antisense orientation, erm
This study
C. botulinum ATCC 3502 cbo0097::intron 84|85S
Insertional disruption of cbo0097 at base 84 in sense orientation, erm
This study
C. botulinum ATCC 3502 cbo0477::intron 152|153AS
Insertional disruption of cbo0477 at base 152 in antisense orientation, erm
This study
C. botulinum ATCC 3502 cbo0477::intron 111|112S
Insertional disruption of cbo0477 at base 111 in sense orientation, erm
This study
C. botulinum ATCC 3502 cbo0558A::intron 121|122AS
Insertional disruption of cbo0558A at base 121 in antisense orientation, erm This study
C. botulinum ATCC 3502 cbo0558A::intron 114|115S
Insertional disruption of cbo0558A at base 114 in sense orientation, erm
This study
Plasmids
pMTL007C-E2
ClosTron plasmid, catP, L1.LtrB intron with ermB RAM, constitutive intron
expression under fdx promoter
University of Nottingham
[77]
pMTL007C-E2:: cbo0097-124|125AS
pMTL007C-E2 with L1.LtrB retargeted to base 124 of cbo0097 in antisense
orientation
This study
pMTL007C-E2:: cbo0097-84|85S
pMTL007C-E2 with L1.LtrB retargeted to base 84 of cbo0097 in antisense
orientation
This study
pMTL007C-E2:: cbo0477-152|153AS
pMTL007C-E2 with L1.LtrB retargeted to base 152 of cbo0477 in sense
orientation
This study
pMTL007C-E2:: cbo0477- 111|112S
pMTL007C-E2 with L1.LtrB retargeted to base 111 of cbo0477 in antisense
orientation
This study
pMTL007C-E2:: cbo0558A-121|122AS
pMTL007C-E2 with L1.LtrB retargeted to base 121 of cbo0558A in sense
orientation
This study
pMTL007C-E2:: cbo0558A- 114|115S
pMTL007C-E2 with L1.LtrB retargeted to base 114 of cbo0558A in antisense
orientation
This study
1ATCC, American Type Culture Collection. doi:10.1371/journal.pone.0089958.t001 1ATCC, American Type Culture Collection. doi:10.1371/journal.pone.0089958.t001 from the foreground signal using the ‘‘normexp’’ method with
offset value of 50 [66]. The Cy3 and Cy5 channels were converted
into a logarithmic (log2) scale and normalized using quantile
normalization [67]. Transcriptomic Analysis with DNA Microarrays The raw microarray expression data collected
at 37uC and 1 h post-shock have been previously deposited in the
Gene Expression Omnibus under accession number GSE26587
[13]. The 5-h raw data and the normalized log2-ratios of
differential transcription 1 h and 5 h after the cold shock
compared with transcription before the shock have been deposited
under accession number GSE51465. Statistical analysis was
performed to find differentially transcribed genes at 1 h and 5 h
after the temperature downshift, in relation to transcript levels
before cold shock. The analysis was done separately for each probe
to control variation within a coding sequence. Moderated t-test
with empirical Bayes variance shrinkage was applied to each probe
on the array and the resulting p-values were converted into false
discovery rate (FDR) values [68]. For each ORF, the probe with
median unmodified p-value for the difference in transcript levels
was chosen to represent the ORF. Of these, ORFs with FDR ,
0.05 were subsequently considered to have a significant difference
in transcript levels. Finally, significantly differentially transcribed
ORFs with a log2 fold change ,22.0 or .2.0 at either time point
were considered to be included in the cold-regulated gene set. from the foreground signal using the ‘‘normexp’’ method with
offset value of 50 [66]. The Cy3 and Cy5 channels were converted
into a logarithmic (log2) scale and normalized using quantile
normalization [67]. The raw microarray expression data collected
at 37uC and 1 h post-shock have been previously deposited in the
Gene Expression Omnibus under accession number GSE26587
[13]. The 5-h raw data and the normalized log2-ratios of
differential transcription 1 h and 5 h after the cold shock
compared with transcription before the shock have been deposited
under accession number GSE51465. Statistical analysis was
performed to find differentially transcribed genes at 1 h and 5 h
after the temperature downshift, in relation to transcript levels
before cold shock. The analysis was done separately for each probe
to control variation within a coding sequence. Moderated t-test
with empirical Bayes variance shrinkage was applied to each probe
on the array and the resulting p-values were converted into false
discovery rate (FDR) values [68]. For each ORF, the probe with
median unmodified p-value for the difference in transcript levels
was chosen to represent the ORF. Of these, ORFs with FDR ,
0.05 were subsequently considered to have a significant difference
in transcript levels. References 11. Dahlsten E, Kirk D, Lindstro¨m M, Korkeala H (2013) Alternative sigma factor
SigK has a role in stress tolerance of group I Clostridium botulinum strain ATCC
3502. Appl Environ Microbiol 79: 3867–3869. 1. Peck MW (1997) Clostridium botulinum and the safety of refrigerated processed
foods of extended durability. Trends Food Sci Technol 8: 186–192. 1. Peck MW (1997) Clostridium botulinum and the safety of refrigerated processed
foods of extended durability. Trends Food Sci Technol 8: 186–192. 1. Peck MW (1997) Clostridium botulinum and the safety of refrigerated processed
foods of extended durability. Trends Food Sci Technol 8: 186–192. 2. Lindstro¨m M, Kiviniemi K, Korkeala H (2006) Hazard and control of group II
(non-proteolytic) Clostridium botulinum in modern food processing. Int J Food
Microbiol 108: 92–104. 2. Lindstro¨m M, Kiviniemi K, Korkeala H (2006) Hazard and control of group II
(non-proteolytic) Clostridium botulinum in modern food processing. Int J Food
Microbiol 108: 92–104. 12. Sebaihia M, Peck MW, Minton NP, Thomson NR, Holden MT, et al. (2007)
Genome sequence of a proteolytic (Group I) Clostridium botulinum strain Hall A
and comparative analysis of the clostridial genomes. Genome Res 17: 1082–
1092. 3. Lindstro¨m M, Korkeala H (2006) Laboratory diagnostics of botulism. Clin
Microbiol Rev 19: 298–314. 13. Dahlsten E, Zhang Z, Somervuo P, Minton NP, Lindstro¨m M, et al. (2014) The
cold-induced two-component system CBO0366/CBO0365 regulates metabolic
pathways with novel roles in cold tolerance of group I Clostridium botulinum ATCC
3502. Appl Environ Microbiol 80: 306–319. 4. den Besten HM, Arvind A, Gaballo HM, Moezelaar R, Zwietering MH, et al. (2010) Short- and long-term biomarkers for bacterial robustness: a framework
for quantifying correlations between cellular indicators and adaptive behavior. PLoS ONE 5: e13746. 14. Ko R, Smith LT, Smith GM (1994) Glycine betaine confers enhanced
osmotolerance and cryotolerance on Listeria monocytogenes. J Bacteriol 176: 426–
431. 5. Phadtare S (2004) Recent developments in bacterial cold-shock response. Curr
Issues Mol Biol 6: 125–136. 6. Kaan T, Homuth G, Mader U, Bandow J, Schweder T (2002) Genome-wide
transcriptional profiling of the Bacillus subtilis cold-shock response. Microbiology
148: 3441–3455. 15. Angelidis AS, Smith LT, Hoffman LM, Smith GM (2002) Identification of
OpuC as a chill-activated and osmotically activated carnitine transporter in
Listeria monocytogenes. Appl Environ Microbiol 68: 2644–2650. 7. Beckering CL, Steil L, Weber MHW, Volker U, Marahiel MA (2002)
Genomewide transcriptional analysis of the cold shock response in Bacillus
subtilis. Acknowledgments The authors wish to thank Marianne Huuskonen, Heidi Ja¨rvima¨ki, Hanna
Korpunen, Esa Penttinen, and Kirsi Ristkari for technical assistance. Transcriptome of C. botulinum in Cold Shock Transcriptome of C. botulinum in Cold Shock 95uC to activate the DNA polymerase, followed by 40 cycles of
denaturation at 95uC for 10 s and annealing and extension at
60uC for 20 s, except for primers cbo0751, cbo1407, cbo2226, and
cbo2847, where the program consisted of an initial heating step of
7 min 95uC, and 40 cycles of denaturation at 95uC for 10 s,
annealing at 55uC for 15 s followed by extension at 72uC for 15 s. The 1:105 diluted no-RT controls were analyzed using the 16S rrn
primers and reaction conditions described above, and no-template
controls were included in each run. cbo0097, cbo0477 and cbo0558A mutants were anaerobically grown
in TPGY broth at 37uC for 12 h or at 17uC for 7 days as described
[11]. The temperature of 17uC was used to avoid the notable
growth variation observed in the wild-type cultures at 15uC, a
temperature close to the minimum growth temperature of C. botulinum ATCC 3502 [79]. The OD600 of the cultures grown at
37uC was measured anaerobically at 1-h intervals, and twice daily
for cultures grown at 17uC. Growth curves were constructed by
plotting the OD600 of each culture against time. The relative quantification of target gene transcript levels at 1 h
after temperature downshift, normalized to reference gene (16S
rrn) transcript levels and calibrated to the samples taken before the
cold shock, were calculated by using the Pfaffl method [72]. The
method takes into account individual primer efficiencies, which
were obtained from RT-qPCR standard curves prepared from
serial dilutions of pooled cDNA samples. The Cq values measured
for 16S rrn remained stable throughout the experiment, supporting
its use as a reliable normalization reference gene in the conditions
studied. 16S rrn is the only previously reported reference gene for
C. botulinum [73,74]; no other suitable reference genes have been
reported. Author Contributions Conceived and designed the experiments: ED PS ML HK. Performed the
experiments: ED PS ML. Analyzed the data: ED MI PS. Wrote the paper:
ED MI PS ML HK. Conceived and designed the experiments: ED PS ML HK. Performed the
experiments: ED PS ML. Analyzed the data: ED MI PS. Wrote the paper:
ED MI PS ML HK. Supporting Information Table S1
Transcript fold changes of C. botulinum
ATCC 3502 1 h and 5 h after temperature downshift
from 37uC to 15uC. Tab 1, All normalized transcript fold
changes; Tabs 2 to 5, k-means clustering of significantly
(FDR,0.05) .2.0 or ,–2.0 log2 fold-affected genes; Tab
6, genes significantly (FDR,0.05) .2.0 or ,22.0 log2
fold-affected at 1 h after cold shock; Tab 7, genes
significantly
(FDR,0.05)
.2.0
or
,22.0
log2
fold-
affected at 5 h after cold shock; Tab 8, functional
categories. Growth Experiments For evaluation of growth at low temperature, three biological
replicate cultures of wild-type C. botulinum ATCC 3502, and Mutant Construction (XLSX) (XLSX) Insertional
knock-out
mutants
for
cbo0097,
cbo0477
and
cbo0558A with intron insertions in two different sites and
orientations were constructed using the ClosTron technology
[75–77]. ClosTron pMTL007C-E2 vectors with de novo synthe-
sized intron targeting regions (DNA2.0 Inc., Menlo Park, CA)
were used for the mutations. The intron insertion sites and
orientations for the mutant strains are presented in Table 1;
primer sequences used to confirm the insertion site and orientation
are presented in Table S2. Single intron insertion was confirmed
by Southern blotting with a probe targeted to the inserted
sequence as described [78]. Table S
(XLSX)
2
Oligonucleotide primers used in the study. RT-qPCR To validate the differences of transcript levels observed in the
DNA microarray experiments, RT-qPCR was performed for
selected genes (cbo0097, cbo0477, cbo0558A, cbo0751, cbo0753,
cbo1323, cbo1407, cbo2226, cbo2227, cbo2304, cbo2525, cbo2847,
cbo2961, cbo3197, and cbo3202) on samples used for microarray
hybridizations. The Cq values of the 1-h post-cold-shock samples
for genes cbo0751, cbo0753, cbo1407, cbo2226, cbo2227, cbo2525,
cbo2847, cbo3199, and cbo3202 were obtained in a previous study
[13]. The Cq values of the pre-cold shock samples for all the
selected genes, and for the 1-h post-cold-shock samples for genes
cbo0097, cbo0477, cbo0558A, cbo1323, cbo2304, and cbo2961, were
obtained in the current study. The DyNAmo Flash SYBR Green
qPCR Kit (Thermo Fisher Scientific) was used according to the
manufacturer’s instructions to set up two replicate qPCR reactions
for each cDNA sample. Each reaction consisted of 1 6 Master
Mix (Thermo Fisher Scientific), 0.5 mM of forward and reverse
primer (Table S2), and 4 ml of 1:20 (cbo0097, cbo0477, cbo0558A,
cbo1323, cbo2304, cbo2961, cbo3197, cbo3202), 1:103 (cbo0751,
cbo0753, cbo1407, cbo2226, cbo2227, cbo2525, cbo2847), or 1:105
(16S rrn) diluted cDNA in a total volume of 20 ml. The reactions
were performed in the Rotor-Gene RG3000 thermal cycler
(Qiagen). The program consisted of an initial heating step of 7 min The identified ORFs were further analyzed by clustering
methods with the R stats and gplots packages. The data were
divided into 4 to 12 clusters by k-means clustering (Euclidean
distance, Hartigan-Wong method [69,70]). Each k-means sub-
cluster was subjected to hierarchical clustering (Euclidean distance,
Ward’s method [71]), and the resulting dendrograms were
visualized as heatmaps. Additionally, a log2 fold change ,21.0
or .1.0 (FDR ,0.05) was accepted for the ORFs belonging to the 11 PLOS ONE | www.plosone.org February 2014 | Volume 9 | Issue 2 | e89958 Transcriptome of C. botulinum in Cold Shock virulence by Clostridium difficile CodY. J Bacteriol 192: 5350–53 26. Cybulski LE, Albanesi D, Mansilla MC, Altabe S, Aguilar PS, et al. (2002)
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virulence by Clostridium difficile CodY. J Bacteriol 192: 5350–5362. Transcriptome of C. botulinum in Cold Shock Stickland LH (1934) Studies in the metabolism of the strict anaerobes (genus
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Clostridium): The chemical reactions by which Cl. sporogenes obtains its energy. Biochem J 28: 1746–1759. February 2014 | Volume 9 | Issue 2 | e89958 13 PLOS ONE | www.plosone.org
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59 Dielectric materials for passive electronic components: from synthesis to processing, a state of the art
I.5 Supercritical fluid synthesis of BaxSr1-xTiO3 (with 0 ≤ x ≤ 1 BST) Only few papers (around 10) report the synthesis of BaTiO3 based materials in supercritical fluids and are mainly shared between two research groups, the supercritical fluid research center at the national institute of Advanced Industrial Science and Technology (AIST – Senday – Japan) and the Supercritical Fluid group at the Institute of Condensed Matter Chemistry of Bordeaux (ICMCB – CNRS – France). From these 10 papers two main approaches are presented towards the synthesis of BaTiO3 based nanoparticles in supercritical fluids: (1) the “supercritical hydrothermal like synthesis” based on hydroxide (Ba(OH)2), oxide (TiO2) or nitrate (Ba(NO3)2) precursors or (2) the “supercritical sol-gel like synthesis” based on alkoxides precursors which not only enable the synthesis of BaTiO3 (as in the case (1)) but also the barium strontium titanate (BST – Ba1-xSrxTiO3 with 0 ≤ x ≤ 1) entire solid solution one. I.5.1 Supercritical hydrothermal synthesis of BaTiO3 (BT) Hayashi et al reported the continuous synthesis of BaTiO3 in supercritical water based on different types of precursors (Ba(NO3)2, Ba(OH)2, [Ba(OH)2, 8H2O] and TiO2). For these syntheses they chose the setup configuration where a preheated line of water is used to heat the media in order to faster the reaction and have a homogeneous nucleation when it is mixed with the precursors at the inlet of the reactor [149,153,155–158]. The precursor line is not preheated to avoid side reactions. The BaTiO3 crystallizes inside the reactor which is under supercritical conditions and the powders are recovered after the back pressure regulator. One type of precursors was a ixture of TiO2 nanoparticles dispersion with a solution of Ba(OH)2. According to the flow rate, the conditions of pressure and temperature, the BT nanoparticles were produced in time ranging from few milliseconds to tens of seconds. The BaTiO3 nanoparticles average size was comprised between 10 to 50 nm; an increase of the mean size increased the size distribution. In the case of 50 nm particle, the size distribution was spread from 10 to 150 nm. These syntheses displayed aggregated particles with high crystallinity. According to the conditions of pressure and temperature they claimed that they change the crystal phase from the cubic to the tetragonal one. To get a tetragonal phase, the temperatures ranged from 400 to 420°C and the pressure from 20 to 35 MPa. They based their investigation on XRD analyses of the powders and more precisely on the full width at half maximum (FWHM) of the XRD peaks at (111) and (200). They estimated that since the FWHM for the (200) peak, characteristic of the tetragonal phase, was larger than the one of the (111) peak, the phase was tetragonal. Nevertheless, we have to be very careful with such interpretation since we do not actually observe the splitting of the 200 and 002 peaks. 60 Dielectric materials for passive electronic components: from synthesis to processing, a state of the art Moreover, based on Figure I-21, the variation of pressure is far too low to have any consequences on the material’s structure. Finally they have not done any ferroelectric characterization to further prove their hypothesis. Two reaction mechanisms similar to the most well-known ones of hydrothermal syntheses were proposed [130]: (1) dissolution of TiO2 precursor followed by crystallization of BaTiO3 as demonstrated by Walton et al [129] or (2) in situ crystallization based on barium diffusion into TiO2 nanocrystals [149,153,155,158]. BaTiO3 was also obtained mixing Ba(NO3)2 with a dispersion of TiO2 nanocrystals and using a residence time of 8 milliseconds leading to nanoparticles of 10 nm with narrow size distribution. The particles were well crystallized in the cubic phase. Again the presence of –OH defects was detected. In this case, the proposed mechanism was based on the hydrolysis and precipitation of the precursors [157]. With the use of these types of precursors only the synthesis of BaTiO3 nanopowders was reported but they did not verify the accuracy of the synthesis measuring their ferroelectric properties and comparing them to the literature. However, as introduced in the following part, the use of alkoxides enables the production of barium strontium titanate nanopowders over the entire solid solution.
I.5.2 Supercritical sol-gel like synthesis of barium strontium titanate
Bocquet et al. were the first to report the synthesis of BaTiO3 starting with a double alkoxide (BaTi(O-iC3H7)6) in supercritical water / isopropanol mixture [151]. In this study, an extra preheated line was used and the temperature was fixed only by the heating system of the reactor. However, because the alkoxides are not stable in water, two lines were used: one for the precursors dissolved in isopropanol, the other for the water. The reactor was divided in two parts: the first one was in vapor phase to hydrolyze the alkoxides and the second one in supercritical conditions to thermally treat the produced nanoparticles. These nanoparticles presented an average size of 10 nm with a narrow size distribution and a specific surface area of 80 to 90 m2/g. The infrared measurements showed –OH defects. Based on these results, Reverón et al. from our group combined both syntheses using alkoxide precursors reacting in water / alcohol mixtures and a preheated line. They first synthesized BaTiO 3 nanopowders and then demonstrated for the first time the synthesis of the entire solid solution of BST nanoparticles in supercritical fluids [159,189,190,209]. 61 Dielectric materials for passive electronic components: from synthesis to processing, a state of the art Barium, strontium and titanium isopropoxides were dissolved in ethanol. The flow reaction system was made of two injection lines: one for the precursors dissolved in ethanol and the second one for water in order to proceed to the hydrolysis of the precursors at the mixing point. The entire system was pressurized at 26 MPa using a back pressure regulating valve. The water line was preheated at 150°C to faster the reaction, however the injection line of precursors was kept at room temperature to avoid side reactions. The two lines were mixed at the inlet of a height meter length tubular reactor with 16 cm3 of inner volume. The first four meters were heated at T1 = 150°C to enhance the hydrolysis of the precursors and the four last meters were heated at T2 = 380°C above the critical point of the water / ethanol mixture to promote the crystallization. Using this setup the experimental parameters were tuned for the BaTiO3 synthesis [159]. One of the most important parameter, beside the pressure and temperature, was the ethanol molar ratio. For an ethanol molar ratio of 0.29 pure BaTiO3 nanoparticles with a crystallinity higher than 90 % were obtained. The particles presented an average size of 41 nm with a size distribution of ± 13 nm. For all the materials the presence of –OH defects was detected. After the optimization of the experimental conditions for BaTiO3 synthesis, the process was transferred to Ba1-xSrxTiO3 (with 0 ≤ x ≤ 1) solid solution synthesis [189,190]. Increasing the strontium amount leads to a decrease of the mean particle size. As shown in Figure I-34 the average size went from 41 ± 13 nm in the case of BaTiO3, to 25±6nm for x = 0.5 and to 19 ± 5nm in the case of SrTiO3. These materials were then uniaxially pressed into pellets and sintered 4 four hours at 1325 °C in order to measure their ferroelectric properties according to their composition (Figure I-34). 62 Dielectric materials for passive electronic components: from synthesis to processing, a state of the art Figure I-34. Effect of barium substitution with strontium on particles size T Curie evolution [189]. The variation of the TCurie according to the strontium content being in agreement with the literature it exhibits the synthesis accuracy in terms of materials composition and stoichiometry [189,209]. However, it is also possible to observe correlation between the composition and the average particles size. Indeed, it appears that the increase of strontium amount tends to decrease the particles size. But so far no precise explanations were found to justify this trend. I.5.3 Conclusion
Although this process enables the synthesis of high quality and well crystallized nanoparticles with narrow size distribution as expected by the industry, there are still just few papers reporting the supercritical synthesis of BaTiO3 or BST nanomaterials. Two main synthesis routes can be used according to the nature of the precursors leading to different reaction mechanisms. The one based on alkoxide precursors enables more flexibility than the other with the possibility to produce the entire solid solution between BaTiO3 and SrTiO3 (Ba1-xSrxTiO3 with 0 ≤ x ≤ 1). In both cases it is important to note that even if the syntheses are achieved under pressure (around 25 MPa) it is too low to have any consequences on the material’s structure (Figure I-21) even if Hakute et al. study claimed that a variation pressure can modify the BaTiO3 crystal structure from cubic to tetragonal phase [155]. It is thus necessary to achieve a deeper study to understand and control the 63 Dielectric materials for passive electronic components: from synthesis to processing, a state of the art nucleation and growth mechanisms, especially in the case of the BST solid solution, where the size variation with the composition remains unexplained and to get a better control on the synthesis. 64 Dielectric materials for passive electronic components: from synthesis to processing, a state of the art I.6 Conclusion of Chapter I
Passive components being space consuming on PCBs, they focus great attention from the industry because they limit the ability to produce more compact and multifunctional electronic devices. The case of capacitors is especially targeted because a large amount of them is necessary in a single device. For example, a cell phone uses hundreds of them to operate. To face this challenge, two types of capacitor configurations are very promising, the MLCCs which are already well known but can be improved and the embedded capacitors which are still at the development stage. In both cases, the operating principle relies on the use of ferroelectric materials such as perovskite BaTiO3 based materials. Such materials have gained a lot of attention from researchers for decades because they present high dielectric permittivity. However the ferroelectricity is very sensitive to the grains size when they reach tens of nanometers. That is why, if we want to maintain a certain level of device performance, it is necessary to use high quality dielectric nanomaterials in terms of size, crystallinity, purity, and homogeneity, while keeping a large amount of production at low cost and following the environmental policy. So far, the main synthesis routes reach their limits and new processes have to be developed. An alternative is the supercritical fluid technology. In addition to bring an answer to the materials quality expectations, this scalable technology enables a fast and reliable materials production using green solvents. Moreover this technology is versatile and not only enables the production of oxide nanoparticles but also metals, nitrides and more complex structures such as core shell ones. The example of the BaTiO3 synthesis is really well illustrating the contribution which can be brought by this technology to the industrial production of metal oxide nanomaterials with the ability to produce in tens of seconds high quality BaTiO 3 based nanoparticles. However, if we want to use it for a large scale production of nanomaterials, it is first necessary to understand their formation in supercritical fluids conditions in order to control their design. Moreover, as already discussed, at the nanoscale it becomes essential to understand the link between the materials structure and their ferroelectric properties. This will be possible with an in depth study of the material structure and surface properties correlated to a characterization of their intrinsic ferroelectric properties. To do that, this PhD work is divided in two parts:
Study of the supercritical fluid synthesis of barium titanate based nanoparticles and its solid solutions; Ba1-xSrxTiO3 with 0 ≤ x ≤ 1 (BST) and BaTi1-yZryO3 with 0 ≤ y ≤ 1 (BTZ) combining ex situ 65 Dielectric materials for passive electronic components: from synthesis to processing, a state of the art analyses like X-ray diffraction, electronic microscopy and different types of spectroscopy with in situ synchrotron powder X-ray diffraction. Process these powders into ceramics for studying either their bulk or intrinsic properties. For the study of bulk properties, we are going to use the “conventional” sintering process enabling the grain growth. However, to study the intrinsic properties it is necessary to make dense and nanostructured ceramics keeping the starting grains size. This is possible using fast sintering methods, such as the spark plasma sintering (SPS) one. This will enable us to improve the knowledge concerning the size effect on their properties and to evaluate their potential application in electronic, especially for capacitors. This part will be conclude with an opening study concerning the development and dielectric characterization of hybrid nanocomposites (nanoparticles / polymer).
66 Dielectric materials for passive electronic components: from synthesis to processing, a state of the art 67
Chapter II. Nanomaterials synthesis, processing and characterization: experimental setups and methods
69 Nanomaterials synthesis, processing and characterization:
experimental
setups and methods 70 Nanomaterials synthesis, processing and
characterization
:
experimental
setups and methods
In the following chapter we are going to present the different methods we used across this PhD, starting with a presentation of the ones to produce and characterize the barium titanate based nanoparticles, where a focus in done on the in situ synchrotron powder diffraction setup enabling to follow in real time the nanoparticles formation in supercritical fluid conditions. Finally, the techniques to process these powders into ceramics or composites and their characterization tools will be described. II.1 Nanoparticles synthesis
To synthesize our particles, we chose to work with a reactor that operates in a continuous mode because it enables to have a good control of the process operating parameters such as pressure, temperature and residence time, while producing a reasonable amount of material to perform both, the characterization and processing steps of the powders. In this first part we will describe the setup and the various characterization tools we used.
II.1.1 Setup
The experimental setup used for this work is based on a former one developed at the ICMCB, in the “supercritical fluids” group (Figure II-1). Indeed it has already been successfully operated to synthesize barium titanate and barium strontium titanate monocrystalline particles [150,159,189,190,209–211]. The strength of this setup lies in its versatility, indeed, such system is quite easy to transform at will.
Figure II-1. Sketch of the experimental setup developed at ICMCB [150,210]
Copyright © 2006, Elsevier. 71 Nano
materials synthesis, processing and characterization: experimental setups and methods
The experimental setup consists in two high pressure pumps (flow up to 15 mL.min-1). For this study, one is dedicated to the injection of the alkoxide precursors solubilized in ethanol, the other one for the injection of water. A third pump can be used equally for the injection of a second solution of metal precursors, a solution of functionalizing agents or even of CO2, depending of the type of study carried out. The injected solutions can also be preheated independently (Ta, Tb, Tc) before being mixed. The pressure is equal in the whole system, verified with manometers (p) and controlled with a back pressure regulator (Tescom®) (see image of the actual setup Figure II-2). For security reasons, security valves adjusted at 40 MPa are located right after the high pressure pumps. Figure II-2. Photography of the continuous flow experimental setup used for the synthesis of nanoparticles in supercritical fluids. The tubular reactor in which the reaction, the nucleation and growth and the crystallization of the nanoparticles take place, is made of stainless steel tube (type 316L, inner diameter of 1.57 mm) and consists in four independent parts of 6 m long each (R1 to R4). This was
made
in
a
view
of
exploring different configurations of reactions (total
length of the reactor
,
split
-up
of
the
hydrolysis and
the
crystallization, etc.). They are maintained
at
constant temperature
with the use
of four heating resistors (Therm
ocoa
x®
)
(T1
to
T
4)
. K
type
thermo
couple
s are positioned all
along the reactor to
enable the
visualization of the temperature profiles
during the synthesis reaction. An insulation
layer minimiz
es
the
heat
radiation of
the reactor
and
the
whole
system
is
put
into
a Plexiglas box
for a safe
protection. The nanoparticles are recovered
either with a
filter
that
blocks
them on the streamline
, or direct
ly in
suspension after the back pressure regulator.
At the outlet of
the reactor
, a part of the circuit is 72 Nanomaterials synthesis, processing and characterization
:
experimental setups and methods deepened in iced water
to
quench the growth of the nanoparticles on the one hand, and on the other hand to prevent the
back pressure regulator
from being overheated and damaged by the hot
fluid
going through
it. The setup is operated and controlled with the help of an interface programmed on the software Labview®.
II.1.2 Fluid hydrodynamic
For such setup, it is of main importance to control the hydrodynamic of the flow within the reactor. First, it enables to adjust the residence time for the reaction, but also, the size distribution of the particles. To do that, we use the Reynolds number (equation (7)) which considers the viscosity and the mass flow of the fluid according to the reactor diameter to define three different regimes: laminar, intermediate or turbulent.
Re = ρ. v. D 4. dm = η π. D. η (7) Where is the density of the fluid (kg.m-3), v the fluid speed (m.s-1), the dynamic viscosity (Pa.s), D the reactor inner diameter (m) and dm the mass flow of the fluid (kg.s-1).
It is important to note that the density and viscosity of the fluid depend on the system temperature and pressure. The density can be estimated with Refprop© developed by the National Institute of Standards and Technology (NIST) and the viscosity with AspenTech© software.
II.1.2.1 Laminar flow
When Re < 2000, the flow is considered as laminar and the speed profile is parabolic (Figure II-3b). The linear speed v (r) (in m.s-1) of an element of fluid located at a distance r (in m) from the axe of the reactor can be expressed as equation (8): 2. r 2 v
(r
) = 2.
v [1
−
(
) ] D (8) The linear speed is then faster on the axis of the tube (r = 0) and is equal to 2 v than on the wall of the reactor where the speed is theoretically equal to zero. The shortest residence time can therefore be estimated from equation (9): 73 Nanomaterials synthesis, processing and characterization: experimental setups and methods ts > L. π. R 2. ρ 2. dm (9) In this case, unlike the plug flow type, the reaction duration will not be constant. It will be faster at the center of the reactor than at its wall. This will directly impact the produced particles size and broader their size distribution.
Figure II-3: a) plug flow, b) laminar flow, c) comparison of
reaction
time with
the type of regime. In addition, because the speed is equal to zero at the side of the reactor, this will favor the deposit of the produced material on the reactor surface. The material having a much longer residence time will grow up to a point where it will be took off from the reactor by the flow. This will lead to the apparition of a secondary population of particles in terms of size.
II.1.2.2 Intermediate flow
For 2000 < Re < 3000, the flow is intermediate between the turbulent and the laminar ones (Figure II-3c), and the Equation (9) is still valid to calculate the shortest residence time. In terms of particles morphology, the size distribution may adopt an intermediate profile too. II.1.2.3 Turbulent flow
When Re > 3000, the flow is considered as turbulent and the reactor can be associated to the plug flow type reactor model (Figure II-3a). The speed and the precursor concentrations are uniform in each “slice” of the fluid and the relation time = distance / speed is valid across the entire reactor section. The residence time can therefore be calculated according to equation (10): L. π. R 2.
ρ ts = dm 74 (10)
Nanomaterials synthesis, processing and characterization: experimental setups and methods
Where ts is the residence time (s), L the length of the reactor (m) and R the inner radius of the reactor (m). The residence time being constant for each particles, they present a narrow size distribution.
II.1.3 Conclusion
Thanks to its versatility, this setup is very interesting because it enables to carry out different types of tests playing on various experimental parameters such as the temperature, pressure, reactor length, multi injection, flow rate and so on, in order to optimize the materials synthesis. One of the critical parameters is the control of the fluid hydrodynamic which mainly impacts the residence time during the synthesis. If it is not constant across the entire reactor section, the produced powders will present a broad size distribution. It is thus preferable, if possible, to carry out the syntheses in a turbulent flow regime to have a plug type reactor, leading to a constant residence time and so to a better control over the size and size distribution of the produced nanoparticles. However it is not the only important one. Changing the pressure, temperature, solvent nature and precursor concentrations can also directly impact the properties of the nanoparticles. In the next section we are going to present the various methods we used to characterize the nanoparticles in terms of structure, morphology, size and surface properties. 75 Nanomaterials synthesis, processing and characterization: experimental setups and methods
II.2 Powder characterization methods
Crossing information from different ex situ and in situ characterization techniques is necessary to describe the produced powders and improve the understanding of mechanisms occurring in the supercritical fluid reactor (Figure II-4).
Figure II-4. Characterization techniques. Many characterization techniques are available at ICMCB and at the “Plateforme Aquitaine de Caractérisation des MATériaux” (PLACAMAT) in Bordeaux or through a collaboration with Prof. B. Iverson at Aarhus University (Denmark) and the following will present the different ones we used, associated with the type of information they provide, to achieve a deep characterization of the produced material.
II.2.1 Powder X-ray diffraction (PXRD)
Across this study, the use of X-ray diffraction is of main interest. We performed both, conventional ex situ powder X-ray diffraction measurements but also in situ synchrotron wide angle X-ray scattering (WAXS) ones using an adapted reactor. 76 Nanomaterials synthesis, processing and characterization: experimental setups and methods
II.2.1.1 Ex situ PXRD measurements
The analyses were carried out by E. Lebraud and S. Pechev from ICMCB internal service, and two types of apparatus were used according to the nature of information we were looking for: A PANalitycal X'pert MPD Bragg-Brentano θ-θ geometry diffractometer equipped with a secondary monochromator over an angular range of 2θ = 8-80°. The Cu Kα1,α2 (λ1 = 1.54060 Å, λ2 = 1.54441 Å) radiations are generated at 40 KV and 40 mA. A PANalytical X’Pert MPD-PRO powder diffractometer equipped with a germanium monochromator to be perfectly monochromatic. We observe only the diffraction of the Cu Kα 1 radiation at λ1 = 1.54060 Å generated at 45 KV and 40 mA. The difference between these two apparatus lies in the quality of the expected XRD patterns. The second being perfectly monochromatic, it is possible to carry out a much more precise analysis than with the first one. Indeed, the first one having the contribution of two wavelengths, leads to the superposition of two XRD patterns mingled at low diffraction angles but which tend to split at higher angles. However, the analysis with the first equipment being much faster (around 30 minutes) compared to the second one (several hours or days), it is very convenient to use it to make a first selection of samples which are then measured with the second one.
II.2.1.2 In situ WAXS measurements
The in situ experiments were carried out in collaboration with Prof. B. Iversen team, especially Dr. K. M. Jensen and E. D. Bøjesen, from Aarhus University (Denmark) at the beamline i711 at MAXII, MAXlab, Lund, Sweden. MAX laboratory has three storage rings (MAX I, MAX II and MAX III) and a fourth one (MAX IV) which is under construction. We used the MAX II ring which is, at the moment, the largest one with a circumference of 90 meters. The electrons are injected in the ring with an energy of 400 MeV and accelerated inside the storage ring. In the case of this ring, the electrons can reach an energy of 1500 MeV. At this energy, the electrons present a speed of 99.99999 % of the light speed and a mass around 300 times higher than their resting mass. The maximum stored current is 300 mA and the number of electrons in circulation is estimated at 500 billion. In the case of this study we used the beamline i711 which enables single crystal and powder diffraction. The beam energy is comprised between 12.6 keV (λ = 0.98 Å) and 8.8 keV (λ = 1.4 Å) with a monochromator selecting photon flux with an energy of 12 keV (λ = 1.03 Å). A four circles
77 Nanomaterials synthesis, processing and characterization: experimental setups and methods diffractometer
Newport kappa is located at 5.5 meters form the monochromator and the detector is a an Oxford Diffraction Titan CCD camera [212,213]. The reactor is a single crystalline sapphire capillary with an internal diameter of 700 μm and a wall thickness comprised between 6 to 80 μm. Such capillary can withstand tens of MPa and is thermally stable up to 450°C while being chemically inert. Finally, the sapphire being made of light elements (α-Al2O3), the X-ray transmission makes possible to observe the diffraction from the particles inside the reactor with a good resolution. As presented in Figure II-5a, the capillary is tighten into a shoe using Swagelok© stainless steel tube fittings and graphite ferrules. The shoe enables to protect the capillary, preventing any external mechanical strain which could break it while under pressure and temperature. It is also possible to insert a thermocouple within the capillary in order to precisely know the temperature during the experiment. The setup is pressurized using a high-performance liquid chromatography (HPLC) high pressure pump, which could reach 40 MPa, connected to the capillary with a 1/16 inch diameter stainless steel tube. The heating is provided by a jet of hot air. The air is preliminary heated at the expected temperature then focused on the capillary using a pneumatic shutter system which controls the direction of the flow (Figure II-5a and b).
Figure II-5. a) Sapphire capillary tighten into the shoe, b) shoe mounted on the heating system and c) top view of the setup [214].
Once
the capillary is mounted into the shoe and pressure tested, it is filled with the precursor mixture using
a syringe. One end of the capillary is then caped and the other connected to the high
78 Nano
material
s synthesis
, processing and
characterization
:
experimental
setups and methods
pressure
pump
. After, the capillary is pressurized using a high pressure pump, and subsequently heated with a jet of hot air. Simultaneously with the onset of the heating, sequential X-ray exposures are initiated (Figure II-6). The synchrotron beam energy was 12.3 keV and powder diffraction patterns were collected with the CCD detector every five seconds. The experiments were allowed to run for at least 20 minutes. Figure II-6. Sketch of the entire system: 1, fastening shoe; 2, thermocouple; 3, beams top; 4, capillary; 5, waste container; 6, heater mouthpiece; 7, solvent; 8, valves (ball-valve and PRV); 9, HPLC pump (including manometer) [214].
It is important to note that, this setup being different than the one used for the flow synthesis described in section II.1, we cannot directly compare the results obtain with each apparatus. The idea is more to compare the trends to get additional insights concerning the materials formation.
II.2.1.3 Interest for this PhD project
The diffraction phenomenon consisting in an interaction between an electromagnetic wave (such as X-ray) with the periodic medium of a crystallized material, the resulting X-ray diffraction (XRD) pattern will depend on the X-ray wavelength and on the internal atomic arrangement intrinsic to the material analyzed. Using the software DIFFRACTplus EVA, it is thus possible to identify the different phases present in the analyzed material comparing the measured diagram with the ones from the powder diffraction files (PDF) reported in the JCPDS-ICDD database. We can then get more precise information on the materials internal structure such as the lattice distortion, the site occupancy or the internal disorder and strain refining the patterns with the software FullProf. 79
Nanomaterials synthesis, processing and characterization: experimental setups and methods
With these profile refinements we can calculate parameters such as the crystallites size using the area below the diffraction peaks. The profile refinements being achieved with the Thomson Cox Hastings – pseudo Voigt approximation, the profiles are described by a convolution of Gaussian (normal distribution, G(x)) and Lorentzian (Cauchy distribution, L(x)) components. Those normalized components (L’(x) and G’(x)) will then give information concerning the peaks broadening (FWHM or H’). Each distribution has a different FWHM: HG and HL and the shape of the Voigt function depend on the relative importance of both contributions, see equation (11): V(x) = V(x, HG, HL ) (11) The pseudo Voigt function, pV(x), approximates the Voigt function where HL and HG are replaced by the pair (η, H), equation (12): pV(x) = ηL′ (x) + (1 − η)G′ (x) with 0 ≤ η ≤ 1 (12) Equation (12) can then be numerically calculated as equation (13): 1/5 H= ( HG 5 + 2.69269. HG 4. HL + 2.48243. HG 3.
HL
2
) +
4.47163.
HG 2. HL 3 + 0.07842. HG. HL 4 + HL 5 (13) With equations (14) and (15) HG = HL = √IG (14) cos(Θ) Y (15) cos(Θ) Leading to equation (16) 5 4 3 1/5 1 √IG + 2.69269. √IG. Y + 2.48243. √IG. Y 2 H= ( ) cos(Θ) + 4.47163. √I 2. Y 3 + 0.07842. √I. Y 4 + Y 5 G G Which can be then inserted
in
the Scherrer formula, equation (17): 80 (16) Nanomaterials synthesis, processing and characterization: experimental setups and methods <D >= K.λ H. cos
(
Θ
)
(17)
The uncertainties
(σD) were calculated with the equations (18) and (19) where σIG and σY are the uncertainties on IG and Y determined through the profile refinement. 0.94λ σD = ( 2 ). σH H σH = σI π Y. √. σIG 2 + G. σY 2 2/3 180 4 Y 16 IG (18) (19)
II.2.2 Transmission electron microscopy (TEM)
Two different apparatus were used: A HITACHI H7650 with an accelerating voltage from 80 to 120 kV which can operate in high contrast or high resolution. Such apparatus enable to have a resolution of 1 nm according to the nature of the material analyzed. In our case, we worked in high resolution mode where the magnification can reach x600 000. The detector is a ORIUS SC1000 11MPx (GATAN) camera and we used the Digital Micrograph (GATAN) software to acquire the images. A JEOL 2200FS with an accelerating voltage of 200 kV and equipped with a high resolution camera. In this case, the resolution is 0.23 nm. Again, the Digital Micrograph (GATAN) software was used to acquire the images. The first TEM is located at the Bordeaux Imaging Center (BIC) on which I was trained and autonomous to do the analyses and the second one is located at PLACAMAT and the high resolution analyses were carried out by S. Buffière. This technique, based on the interaction between the electrons from the microscope beam going through the sample and the ones from the sample itself to create an image, enables to observe the nanoparticles in terms of size (to determine their size distribution) and morphology. It is also possible, in high resolution mode (HRTEM) to observe the atomic arrangement. This enables to discuss the crystallite size calculated from the XRD measurements and see if the nanoparticles are monocrystalline. Finally, such analysis gives information concerning the homogeneity of the powder. 81
Nanomaterials synthesis, processing and characterization: experimental setups and methods II.2.3 Spectroscopy II.2.3.1 Raman spectroscopy
The Raman used is a DXR dispersive microscope with a 532 nm wavelength excitation laser operating at 8 mW output power. The focus is made with a confocal microscope with a x10 objective, leading to a surface analysis of 3.5 μm2. The resolution was 3 cm-1 over a window from 15 to 3550 cm-1. The Raman spectroscopy was very important in the case of our study. Indeed, as presented in the previous chapter (section I.2.2.2) for the BaTiO3 case, at the nanometer scale, the materials structure is modified due to the surface strain leading to a pseudo cubic structure. Contrary to the XRD analysis, which represents an average response of the materials diffraction, the Raman technique enables a much more local analysis. It is a very sensitive tool to characterize crystal structures and phase transitions. Measuring the vibration frequency of a crystal lattice or a molecule (phonons), it detects the local dynamic symmetry in small regions (coherence length lower than 2nm) making it useful to prove non centrosymmetric local structure in an apparent pseudo-cubic nanopowders.
II.2.3.2 Fourier transform infrared spectroscopy (FTIR)
The FTIR measurements were carried out with a Bruker Equinox 55 spectrophotometer. The absorption spectra were acquired through 32 scans in a window from 400 to 4000 cm −1, with a resolution of 4 cm-1. Similarly to the Raman analysis, through the vibration of atomic bonds, it is possible to identify the materials fingerprint and so its purity. However, in the present study it is mostly interesting to use it in order to characterize the surface properties of our materials. II.2.3.3 X-ray photoelectron spectrometry (XPS)
Two different apparatus were used in this study: ESCALAB VG 220i-XL using Mg Kα as X-Ray source of 1253.6 eV which is not monochromatic. The vacuum chamber was set at 10−7 Pa and the analyzed area is a disc with a diameter of 150 μm. This equipment only compensates the electronic charges. K-ALPHA using Mg Kα as X-Ray source of 1486.6 eV which is monochromatic. The vacuum chamber was set at 10−7 Pa and the analyzed area was a disc with a diameter of 200 μm. This equipment which compensates both, the electronic and ionic charges offers a better resolution.
82
Nanomaterials synthesis, processing and characterization: experimental setups and methods
In both case the measurements were carried out by C. Labrugère at the PLACAMAT platform and the spectra were analyzed with the software AVANTAGE (ThermoFisher Scientific). This technique enables an analysis of the materials surface properties through measurement of the kinetic energy and number of electrons released after the atomic ionization due to a X-ray exposition. Making it complementary to the observation we can do using the FTIR spectroscopy.
II.2.4 Conclusion
Combining these different types of analyses, such as XRD, TEM, Raman, FTIR and XPS, which have different levels of resolution enables to make complete analysis of the material. In addition, to characterize the structure and surface properties of the nanoparticles, we can follow in real time their formation in supercritical conditions with the in situ WAXS setup. Once the particles are produced and characterized, the next step is to study the ferroelectric properties to first improve the basic knowledge and then, see if such material can answer the industrial demand in terms of properties. To do these measurements we first have to densify the powders into ceramics. 83
Nanomaterials synthesis, processing and characterization: experimental setups and methods II.3 Powders processing and characterizations II.3.1 Ceramics
Performing ceramics will allow us to determine dielectric properties of the ferroelectric nanopowders synthesized by supercritical fluids. The choice of the sintering process depends on the aimed objectives: reaching the highest density, keeping the initial grain size of the nanopowder, controlling composition and defects. There is an intimate link between the crystal structure of the particles, the nano/microstructure of the ceramics and their dielectric properties. It is thus possible in ceramics to correlate the microstructural and dielectric features. The dielectric characteristics depend on the porosity (which decreases the dielectric permittivity), the grain size (which can affect the transition temperature, the permittivity and losses values), thus the density and the quality of grain boundaries which can be considered as defects rich regions. Besides, by its ability to act at several scales, the ceramic technology can be a key strategy. Through different adjustable parameters (such as the temperature, atmosphere, heating and cooling rates), it is possible to control during the thermal treatment the diffusion process, composition gradients, grain growth and defects chemistry. According to the solid solution studied and to our objectives, we used either; conventional sintering or spark plasma sintering (SPS). The efficiency of SPS to sinter nanomaterials has been fully demonstrated. In this fast sintering process the combination of pulsed electric current and uniaxial pressure leads to efficient heat transfer. As a result, sintering kinetics are increased. In addition, temperature and duration treatment can be significantly reduced compared to conventional sintering limiting the available time for grain growth. This work was achieved in close collaboration with Dr. C. Elissalde team at ICMCB and Dr. C. Estournes team at the “Centre Interuniversitaire de Recherche et d’Ingénierie des Matériaux Plateforme Nationale CNRS de Frittage Flash” (PNF2 / CIRIMAT) from the University Paul Sabatier in Toulouse (France).
II.3.1.1 II.3.1.1.1 Powders sintering
Conventional sintering The first step to make a pellet is to carefully mill the powder in order to break as much as possible the aggregates which could later on lead to inhomogeneity and further differential sintering within the ceramic. A binder, which is a polymer is added (few weight % of the powder) to enhance the 84 Nanomaterials synthesis, processing and characterization: experimental setups and methods mechanical strength of the raw pellet. The mixture is then poured in 8 mm diameter die and uniaxially pressed with a pressure around 1 T/cm2 using a hydraulic press. The pellet is then sintered in a furnace under air atmosphere. A first temperature dwell is realized at few hundreds degrees (around 500°C) for one hour in order to gently burn out the binder used to make the pellet and let the powder self-reorganize to fill the voids created from the binder. Then the temperature is increased up to 1200°C or more for several hours (at least four hours) in order to densify the ceramic through a grain growth before cooling the furnace down. In such process, the heating and cooling rates are generally around 100 to 300°C/h and the overall cycle takes several hours. II.3.1.1.2 Spark plasma sintering (SPS)
As already mention, in comparison to the conventional sintering process, for which the main limitation for nanopowders consolidation is the duration of the thermal cycle, SPS enables very high heating rates and short dwell times of few minutes, leading to enhanced sintering kinetics. Using this technique it is possible to make dense materials keeping the initial grain size of the nanoparticles enabling thus to determine size effect, grain boundaries and defects influences on the final dielectric properties of the ceramics. The powder (without any binder) is loaded in a graphite die, an uniaxial pressure is applied and a pulsed electrical current is allowed to pass via electrodes through the die. The heating source is not external but the die acts as a heating source (Figure II-7). The sintering is generally carried out under reducing conditions (low oxygen partial pressure). A post annealing in air is usually performed to remove surface carbon contamination and to eliminate oxygen vacancies if necessary. However it is possible to work in inert atmosphere (argon, nitrogen) and in some very specific cases in air as it is actually in development in CIRIMAT –PNF2 Toulouse.
Figure II-7. Sketch and image of a SPS apparatus.
85 Nanomaterials synthesis, processing and characterization: experimental setups and methods
Similarly to conventional furnace, it is possible to adjust the sintering atmosphere. However, the difference lies in the heating principle. In this case it is brought by a high electric current going through the die. This current can be either continuous or pulsed. This enables to quickly reach high temperatures (hundreds of °C.min-1) while applying high pressure (hundreds of MPa) leading to sintering cycles of few minutes which is much shorter than the conventional case. We had access to two different SPS apparatus to carry out the different experiments: At the ICMCB we used a Dr. Sinter 515S machine manufactured by the society SPS Syntex Inc and operated by Dr. U-C. Chung. This apparatus has an output pulsed current of 1500 A, a voltage ranging from 2 to 20 V and can apply a pressure up to 200 MPa. At the “Centre Interuniversitaire de Recherche et d’Ingénierie des Matériaux Plateforme Nationale CNRS de Frittage Flash” (PNF2 / CIRIMAT) from the University Paul Sabatier in Toulouse (France) in a collaboration with Dr. C. Estournes team and, especially G. Chevalier and Dr. R. Epherre. The equipment is a Dr Sinter 2080 manufactured by the society SPS Syntex Inc. which can deliver a pulsed current of 8000 A with a voltage of 15 V and can apply a pressure up to the GPa.
II.3.1.2 Ceramic characterization
Similarly to the powder characterization, XRD is the first analysis to be performed on the ceramic in order to see if there are any variations in terms of purity, composition and grain size resulting from the sintering. Then, it is also necessary to determine the ceramics density, using a pycnometer. Dielectric measurements and microstructural studies using scanning electron microscopy (SEM) are then performed. II.3.1.2.1 Dielectric measurements
The measurements were carried out with an automatic Wayne-Kerr 6425 impedance bench developed at ICMCB in a frequency range from 100 Hz to 200 kHz. Gold electrodes are deposited on two parallel faces of the ceramic and the real and imaginary contributions of the permittivity; Ɛ’ and Ɛ”, are respectively determined from the measured values of capacitance (C) and losses (tan δ) from equations (20) and (21): Ɛ′ = C. e Ɛ0. S
Ɛ" =
Ɛ
′. tan
δ 86 (20) (21)
Nanomaterials synthesis, processing and characterization: experimental setups and methods
Where Ɛ0 is the vacuum permittivity (Ɛ0 = 8.854187.10-12 F.m-1), S is the surface of the electrodes (in m2), e the thickness of the dielectric material (in m). The contribution from the measurement apparatus is removed through its calibration in open circuit. The first step is the deposit of thin layer of gold on the ceramic, using physical vapor deposition, to make the electrodes. Then we used the setup described in Figure II-8.
Figure II-8. a) sketch of the dielectric measurement cell, b) image of the open cell and c) image of the closed cell. After the calibration the ceramic is placed between the two nickel electrodes in a quartz cell to control the atmosphere during the measurements. It can either be vacuum or helium, which is more conductive, enabling to start the measurements from a lower temperature (≈ 100 K). The sample is preliminary heated up to 150°C to evaporate, and absorbed with a nitrogen trap, any traces of water which can parasite the measurements. The cell is then cooled down putting it into a liquid nitrogen bath and progressively heated up using a tubular oven mounted on it, with a heating rate of 1°C.min1. The measurements are performed for different frequencies while the sample is heated. For this work, the characterization setup was mainly operated by M. Albino and J. Macaigne at ICMCB.
II.3.1.2.2 Microstructure analysis with scanning electron microscopy (SEM)
There were four apparatus accessible: (1) a JEOL 6360A at ICMCB with an accelerating voltage from 0.5 to 30 kV and a magnification up to 30000 operated by M. Albino, (2) a JEOL JSM 6700F which is a high resolution one with an accelerating voltage from 0.5 to 30 kV and magnification up to 650000 at PLACAMAT and operated by S. Buffière, (3) another JEOL JSM 6700F operated by A. Weibel and A. Peigney at the Toulouse center of micro characterization(UMS Raimond Castaing) and (4) a JEOL JSM 87 Nanomaterials synthesis, processing and characterization: experimental setups and methods 7800F Prime which is a high resolution one with an accelerating voltage from 1 to 30 kV and magnification up to 1000000 6700F operated by A. Weibel and A. Peigney at the Toulouse center of micro characterization(UMS Raimond Castaing). This technique, based on the interaction between the electrons from the microscope beam and the sample enables to observe the microstructure and homogeneity of ceramics in terms of grains size, composition and porosity. This is possible through different types of electron – material interactions: Secondary electrons providing information concerning the material morphology. Backscattered electrons, giving a chemical contrast which enables to observe the ceramics homogeneity in term of composition. Indeed, the lighter elements providing less backscattered electrons, appear darker. Photon X, with characteristic energies for each element, enables also to see the homogeneity within the ceramic. This analysis is known as energy dispersive spectrometry (EDS).
II.3.2 Nanocomposites
For this part of the study, the processing and characterization of the composites were performed in collaboration with Dr. I. Bord-Majek and M. Wade at the institute of Integration Materials to System in Bordeaux (IMS CNRS) and with Prof. R. Jakoby, Dr. Yuliang Zheng and L. Donghang at TU Darmstadt in the Institute for Microelectronic and Photonic (Germany).
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Development and Positioning Accuracy Assessment of Single-Frequency Precise Point Positioning Algorithms by Combining GPS Code-Pseudorange Measurements with Real-Time SSR Corrections
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English
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article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).
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Two new species of the genus Opopaea (Araneae, Oonopidae) from Myanmar
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Yanfeng Tong
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Copyright Yanfeng Tong et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC
BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. * Contributed equally as the first authors. Keywords Goblin spider, morphology, new species, taxonomy Two new species of the genus Opopaea
(Araneae, Oonopidae) from Myanmar Yanfeng Tong1,2*, Zengliang Chen3*, Shuqiang Li4 1 Life Science College, Shenyang Normal University, Shenyang 110034, China 2 Southeast Asia Biological
Diversity Research Institute, Chinese Academy of Sciences, Yezin, Nay Pyi Taw 05282, Myanmar 3 The Seri
cultural Research Institute of Liaoning Province, Fengcheng, Liaoning 118100, China 4 Institute of Zoology,
Chinese Academy of Sciences, Beijing 100101, China Corresponding author: Shuqiang Li (lisq@ioz.ac.cn) emic editor: D. Dimitrov | Received 29 November 2019 | Accepted 11 February 2020 | Published 9 March 202 http://zoobank.org/554574CC-B3C0-49EB-B099-8B34E80E39C6 http://zoobank.org/554574CC-B3C0-49EB-B099-8B34E80E39C6 Citation: Tong Y, Chen Z, Li S (2020) Two new species of the genus Opopaea (Araneae, Oonopidae) from Myanmar. ZooKeys 917: 51–61. https://doi.org/10.3897/zookeys.917.48924 51
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ZooKeys 917: 51–61 (2020)
doi: 10.3897/zookeys.917.48924
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ZooKeys 917: 51–61 (2020)
doi: 10.3897/zookeys.917.48924
http://zookeys.pensoft.net (Araneae, Oonopidae) f
RESEARCH ARTICLE Abstract Two new species of the genus Opopaea Simon, 1892 are reported from Myanmar, O. kanpetlet Tong & Li,
sp. nov. (♂♀) and O. zhigangi Tong & Li, sp. nov. (♂♀). Morphological descriptions and photographic
illustrations of the two new species are given. All types are preserved in the Institute of Zoology, Chinese
Academy of Sciences in Beijing (IZCAS). Introduction The genus Opopaea Simon, 1892 is a widespread and highly diverse goblin spider ge
nus, with biodiversity hotspots in Africa, Asia and Australia (Baehr 2013). A total of
185 valid extant species are currently known, in which 46 in Africa, 33 in Asia, 96 in
Australia and New Caledonia, and 10 in other areas (WSC 2020). The genus Opopaea of Myanmar and the adjacent countries has been poorly stud
ied. Brignoli (1978) reported a new species from Bhutan. Tong and Li (2013) reported Yanfeng Tong et al. / ZooKeys 917: 51–61 (2020) 52 two new species and one newly recorded species from Laos. There are no records of
the genus Opopaea in Myanmar. However, four species of the genus Gamasomorpha
Karsch, 1881 and two recently described species of the endemic genus Kachinia Tong
& Li, 2018 have been reported from Myanmar (Tong and Li 2018; WSC 2020). The
present paper expands the known oonopid diversity of Myanmar by adding one newly
recorded genus and two new species. Materials and methods Used in
the figures: apo = apodeme; asr = anterior scutal ridge; boc = booklung covers; cb =
cymbiobulbus; dte = dorsolateral, triangular extensions; fm = femur; fn = fenestra; ga =
globular appendix; nlp = nail-like process; pd = postgynal depression; pls = paddle-like p
g
p
y
= posterior median eyes; PME-PME = distance between PME and PME; PLE-PME
= distance between PLE and PME; PTL = patella length; TL = total length. Used in
the figures: apo = apodeme; asr = anterior scutal ridge; boc = booklung covers; cb =
cymbiobulbus; dte = dorsolateral, triangular extensions; fm = femur; fn = fenestra; ga =
globular appendix; nlp = nail-like process; pd = postgynal depression; pls = paddle-like
sclerite; psr = posterior scutal ridge; pt = patella; sr = scutal ridge. = posterior median eyes; PME-PME = distance between PME and PME; PLE-PME
= distance between PLE and PME; PTL = patella length; TL = total length. Used in
the figures: apo = apodeme; asr = anterior scutal ridge; boc = booklung covers; cb =
cymbiobulbus; dte = dorsolateral, triangular extensions; fm = femur; fn = fenestra; ga =
globular appendix; nlp = nail-like process; pd = postgynal depression; pls = paddle-like
sclerite; psr = posterior scutal ridge; pt = patella; sr = scutal ridge. Materials and methods The specimens were examined in 95% ethanol using a Leica M205C stereomicroscope. Details were studied with an Olympus BX51 compound microscope. Photos were
taken with a Canon EOS 750D zoom digital camera (18 megapixels) mounted on an
Olympus BX51 compound microscope. Scanning electron microscope images (SEM)
were taken under high vacuum with a Hitachi TM3030 after critical point drying and
gold-palladium coating. All measurements were taken using an Olympus BX51 com
pound microscope and are given in millimeters in the text. The materials are preserved
in the Institute of Zoology, Chinese Academy of Sciences in Beijing (IZCAS). gy
y
j g
Terminology mainly follows Andriamalala and Hormiga (2013). The following
abbreviations are used in the text: AL = abdomen length; ALE = anterior lateral eyes;
ALE-ALE = distance between ALE and ALE; ALE-PLE = distance between ALE and
PLE; AW = abdomen width; CBL = cymbiobulbus length; CBW = cymbiobulbus
width; CL = carapace length; CW = carapace width; EGW = eye group width; FI =
femur insertion on patella; FML = femur length; PLE = posterior lateral eyes; PME
= posterior median eyes; PME-PME = distance between PME and PME; PLE-PME
= distance between PLE and PME; PTL = patella length; TL = total length. Used in
the figures: apo = apodeme; asr = anterior scutal ridge; boc = booklung covers; cb =
cymbiobulbus; dte = dorsolateral, triangular extensions; fm = femur; fn = fenestra; ga =
globular appendix; nlp = nail-like process; pd = postgynal depression; pls = paddle-like
sclerite; psr = posterior scutal ridge; pt = patella; sr = scutal ridge. Terminology mainly follows Andriamalala and Hormiga (2013). The following
abbreviations are used in the text: AL = abdomen length; ALE = anterior lateral eyes;
ALE-ALE = distance between ALE and ALE; ALE-PLE = distance between ALE and
PLE; AW = abdomen width; CBL = cymbiobulbus length; CBW = cymbiobulbus
width; CL = carapace length; CW = carapace width; EGW = eye group width; FI =
femur insertion on patella; FML = femur length; PLE = posterior lateral eyes; PME
= posterior median eyes; PME-PME = distance between PME and PME; PLE-PME
= distance between PLE and PME; PTL = patella length; TL = total length. Taxonomy Family Oonopidae Simon, 1890
Genus Opopaea Simon, 1892 Opopaea kanpetlet Tong & Li, sp. nov. http://zoobank.org/DABDCD9E-6129-4704-A835-E64C97836930
Figures 1–3, 7A–C Type material. Holotype: ♂ (IZCAS Ar-25098), sifting leaf litter, Myanmar, Chin,
Roadside between Kanpetlet and Nat Ma Taung National Park, 003, 21°13.325'N, New Opopaea (Araneae, Oonopidae) from Myanmar New Opopaea (Araneae, Oonopidae) from Myanmar 53 p p
(
p
) f
y Figure 1. Opopaea kanpetlet sp. nov., male holotype A, C, E habitus, dorsal, ventral and lateral views
B, D, F, G prosoma, dorsal, ventral, lateral and anterior views H, I abdomen, anterior and ventral views
J, K, L left palp, prolateral, retrolateral and dorsal views. Abbreviations: boc = booklung covers; dte =
dorsolateral, triangular extensions; sr = scutal ridge. Scale bars: 0.4 mm (A–I); 0.2 mm (J–L). Figure 1. Opopaea kanpetlet sp. nov., male holotype A, C, E habitus, dorsal, ventral and lateral views
B, D, F, G prosoma, dorsal, ventral, lateral and anterior views H, I abdomen, anterior and ventral views
J, K, L left palp, prolateral, retrolateral and dorsal views. Abbreviations: boc = booklung covers; dte =
dorsolateral, triangular extensions; sr = scutal ridge. Scale bars: 0.4 mm (A–I); 0.2 mm (J–L). Yanfeng Tong et al. / ZooKeys 917: 51–61 (2020)
54
Figure 2. Opopaea kanpetlet sp. nov., holotype, left male palp, SEM A, B prolateral and retrolateral views
C, D, G cymbiobulbus, prolateral, retrolateral and dorsal views E, F, H distal part of cymbiobulbus, prolat
eral, retrolateral and dorsal views. Abbreviations: cb = cymbiobulbus; fm = femur; fn = fenestra; pt = patella. Yanfeng Tong et al. / ZooKeys 917: 51–61 (2020) 54 Figure 2. Opopaea kanpetlet sp. nov., holotype, left male palp, SEM A, B prolateral and retrolateral views
C, D, G cymbiobulbus, prolateral, retrolateral and dorsal views E, F, H distal part of cymbiobulbus, prolat
eral, retrolateral and dorsal views. Abbreviations: cb = cymbiobulbus; fm = femur; fn = fenestra; pt = patella. Figure 2. Opopaea kanpetlet sp. nov., holotype, left male palp, SEM A, B prolateral and retrolateral views
C, D, G cymbiobulbus, prolateral, retrolateral and dorsal views E, F, H distal part of cymbiobulbus, prolat
eral, retrolateral and dorsal views. Abbreviations: cb = cymbiobulbus; fm = femur; fn = fenestra; pt = patella. New Opopaea (Araneae, Oonopidae) from Myanmar New Opopaea (Araneae, Oonopidae) from Myanmar 55 Figure 3. Opopaea kanpetlet sp. Taxonomy nov., female (IZCAS Ar-25099) A, C, E habitus, dorsal, ventral and
lateral views B, D, F prosoma, dorsal, ventral and lateral views. Scale bars: 0.4 mm. Figure 3. Opopaea kanpetlet sp. nov., female (IZCAS Ar-25099) A, C, E habitus, dorsal, ventral and
lateral views B, D, F prosoma, dorsal, ventral and lateral views. Scale bars: 0.4 mm. 93°55.739'E, 2942 m, 30.IV.2017, Wu J & Chen Z. Paratypes: 2♀ (IZCAS Ar-
25099, 25100), same data as holotype; 1♂ (IZCAS Ar-25101), 3♀ (IZCAS Ar-25102,
25103, 25104), sifting leaf litter, Myanmar, Chin, near 16.5 km of the roadside be
tween Kanpetlet and Nat Ma Taung National Park, 002, 21°13.195'N, 93°16.125'E,
2789 m, 30.IV.2017, Wu J & Chen Z. 93°55.739'E, 2942 m, 30.IV.2017, Wu J & Chen Z. Paratypes: 2♀ (IZCAS Ar-
25099, 25100), same data as holotype; 1♂ (IZCAS Ar-25101), 3♀ (IZCAS Ar-25102,
25103, 25104), sifting leaf litter, Myanmar, Chin, near 16.5 km of the roadside be
tween Kanpetlet and Nat Ma Taung National Park, 002, 21°13.195'N, 93°16.125'E,
2789 m, 30.IV.2017, Wu J & Chen Z. Etymology. The specific name is a noun in apposition taken from the type lo Diagnosis. The new species is similar to Opopaea tumida Tong & Li, 2013, but
can be distinguished by the small booklung covers (Fig. 1I), the acute tip of male palpal
bulb (Figs 1J–L, 2) and the posterior scutal ridge of the female (Fig. 7A–C). The male
of O. tumida has large booklung covers, a small apophysis in the retrolateral distal re
gion of the palpal bulb, and the female is lacking the posterior scutal ridge (Tong and
Li 2013: figs 8I, 9I, J, 10D). Diagnosis. The new species is similar to Opopaea tumida Tong & Li, 2013, but
can be distinguished by the small booklung covers (Fig. 1I), the acute tip of male palpal
bulb (Figs 1J–L, 2) and the posterior scutal ridge of the female (Fig. 7A–C). The male
of O. tumida has large booklung covers, a small apophysis in the retrolateral distal re
gion of the palpal bulb, and the female is lacking the posterior scutal ridge (Tong and
Li 2013: figs 8I, 9I, J, 10D). Description. Male (holotype). Measurements: TL: 1.63; CL: 0.68; CW: 0.58;
AL: 0.91; AW: 0.69; ALE: 0.08; PME: 0.07; PLE: 0.06; EGW: 0.23; ALE-ALE:
0.04; ALE-PLE: 0.01; PME-PME: 0; PLE–PME: 0; CBL: 0.23; CBW: 0.08; PTL:
0.33; FI: 0.16; FML: 0.13. Taxonomy Coloration: legs yellow, carapace and abdomen scuta yel Yanfeng Tong et al. / ZooKeys 917: 51–61 (2020) 56 low brown, abdominal interscutal areas creamy white, booklung covers light yellow,
pedipalps reddish brown. Habitus as in Fig. 1A, C, E. Carapace (Fig. 1B, F): wide
oval in dorsal view; sides with longitudinal streaks; median area smooth with rows of
setae at lateral edges. Eyes (Fig. 1B, G): ALE largest, PLE smallest; posterior eye row
recurved viewed from above, procurved from front; ALE separated by less than their
radius, ALE–PLE separated by less than ALE radius, PME touching throughout most
of their length, PLE–PME separated by less than PME radius. Clypeus height about
1.1 times ALE diameter (Fig. 1G). Sternum (Fig. 1D) longer than wide, fused to car
apace; surface smooth; radial furrows present between coxae I-II, II-III, III-IV, with
rows of small pits. Endites anteriorly with a small, sharply pointed projection. Abdo
men: booklung covers very small, ovoid, without setae. Pedicel tube short, ribbed,
with small, dorsolateral triangular extensions, scuto-pedicel region lower than pedicel
diameter, with arched scutal ridges, interrupted medially, with curved anterior scutal
ridge (Fig. 1H, I). Palp (Figs 1J–L, 2): femur slightly shorter than half length of
patella and submedially attached to patella; patella strongly enlarged, elongate oval;
tibia small, rounded; cymbiobulbus shorter than the patella; palpal fenestra small
oval and located nearly at tip of cymbiobulbus. Tip of embolus acute triangle. Female (n = 5). As in male except as noted. Measurements (IZCAS Ar-25099):
TL: 1.89; CL: 0.70; CW: 0.61; AL: 1.27; AW: 0.75; ALE: 0.08; PME: 0.06; PLE:
0.05; EGW: 0.21; ALE-ALE: 0.03; ALE-PLE: 0.01; PME-PME: 0; PLE-PME: 0. Palp light yellow. Habitus as in Fig. 3A, C, E. Endites without projections. Copula
tory organ (Fig. 7A–C): posterior margin of epigastric scutal ridge (asr) smooth, thick
posterior scutal ridge (psr) adjacent to asr, small postgynal semicircular depression (pd)
between asr and psr; dorsally with nail-like process (nlp) connected to paddle-like scle
rite (pls) bearing thin, straight arms. Distribution. Known only from the type locality. Opopaea zhigangi Tong & Li, sp. nov. http://zoobank.org/21168390-94EC-4387-9CA9-3ADF1526DEE7
Figures 4–6, 7D–F Opopaea zhigangi Tong & Li, sp. nov. http://zoobank.org/21168390-94EC-4387-9CA9-3ADF1526DEE7
Figures 4–6, 7D–F Type material. Holotype: ♂ (IZCAS Ar-25105), sifting leaf litter, Myanmar, Chin,
near 1.5 km of the roadside between Kanpetlet and Nat Ma Taung National Park,
011–012, 21°13.058'N, 93°59.033'E, 2421 m, 1.V.2017, Wu J & Chen Z. Taxonomy Paratype:
1♀ (IZCAS Ar-25106), same data as holotype. Etymology. The specific name is after Mr Zhigang Chen, one of the collectors of
this species; noun (name) in genitive case. p
g
Diagnosis. The new species is similar to Opopaea deserticola Simon, 1892, but can
be distinguished by the longer palpal patella (the ratio of width to length about 0.5)
and slender cymbiobulbus (the ratio of width to length about 0.35) of male (Figs 4J–L,
5) and very small postgynal depression of female (Fig. 7E). The male of O. deserticola
has relatively shorter palpal patella (the ratio of width to length about 0.65) and ex New Opopaea (Araneae, Oonopidae) from Myanmar New Opopaea (Araneae, Oonopidae) from Myanmar 57 New Opopaea (Araneae, Oonopidae) from Myanmar
57
Figure 4. Opopaea zhigangi sp. nov., male holotype A, C, E habitus, dorsal, ventral and lateral views
B, D, F, G prosoma, dorsal, ventral, lateral and anterior views H, I abdomen, anterior and ventral views
J, K, L left palp, prolatral, retrolateral and dorsal views. Abbreviations: boc = booklung covers; dte = dor
solateral, triangular extensions; sr = scutal ridge. Scale bars: 0.4 mm. Figure 4. Opopaea zhigangi sp. nov., male holotype A, C, E habitus, dorsal, ventral and lateral views
B, D, F, G prosoma, dorsal, ventral, lateral and anterior views H, I abdomen, anterior and ventral views
J, K, L left palp, prolatral, retrolateral and dorsal views. Abbreviations: boc = booklung covers; dte = dor
solateral, triangular extensions; sr = scutal ridge. Scale bars: 0.4 mm. Yanfeng Tong et al. / ZooKeys 917: 51–61 (2020) 58 Yanfeng Tong et al. / ZooKeys 917: 51–61 (2020)
58
Figure 5. Opopaea zhigangi sp. nov., holotype, left male palp, SEM A, B prolateral and retrolateral views
C, D, G cymbiobulbus, prolateral, retrolateral and dorsal views E, F, H distal part of cymbiobulbus, prolat
eral, retrolateral and dorsal views. Abbreviations: cb = cymbiobulbus; fm = femur; fn = fenestra; pt = patella. Figure 5. Opopaea zhigangi sp. nov., holotype, left male palp, SEM A, B prolateral and retrolateral views
C, D, G cymbiobulbus, prolateral, retrolateral and dorsal views E, F, H distal part of cymbiobulbus, prolat
eral, retrolateral and dorsal views. Abbreviations: cb = cymbiobulbus; fm = femur; fn = fenestra; pt = patella. New Opopaea (Araneae, Oonopidae) from Myanmar 59 p p
p
f
y
Figure 6. Opopaea zhigangi sp. panded cymbiobulbus (the ratio of width to length about 0.47) and the female has a
relatively larger postgynal depression (Platnick and Dupérré 2009: figs 55–66).
Description. Male (holotype). Measurements: TL: 1.54; CL: 0.60; CW: 0.58;
AL: 0.98; AW: 0.70; ALE: 0.08; PME: 0.07; PLE: 0.07; EGW: 0.22; ALE–ALE: 0.02;
ALE–PLE: 0.01; PME–PME: 0; PLE–PME: 0; CBL: 0.31; CBW: 0.11; PTL: 0.41;
FI: 0.21; FML: 0.14. Coloration: legs yellow, carapace and abdomen scuta brown,
abdominal interscutal areas creamy white, booklung covers reddish, pedipalps reddish
brown. Habitus as in Fig. 4A, C, E. Carapace (Fig. 4B, F): wide oval in dorsal view;
sides with longitudinal streaks; median area smooth with some setae at lateral edges.
Eyes (Fig. 4B, G): ALE largest, PLE smallest; posterior eye row recurved viewed from
above, procurved from front; ALE almost touching, ALE–PLE separated by less than
ALE radius, PME touching throughout most of their length, PLE–PME separated
by less than PME radius. Clypeus height about 1.7 times ALE diameter (Fig. 4G). panded cymbiobulbus (the ratio of width to length about 0.47) and the female has a
relatively larger postgynal depression (Platnick and Dupérré 2009: figs 55–66).
Description. Male (holotype). Measurements: TL: 1.54; CL: 0.60; CW: 0.58;
AL: 0.98; AW: 0.70; ALE: 0.08; PME: 0.07; PLE: 0.07; EGW: 0.22; ALE–ALE: 0.02;
ALE–PLE: 0.01; PME–PME: 0; PLE–PME: 0; CBL: 0.31; CBW: 0.11; PTL: 0.41;
FI: 0.21; FML: 0.14. Coloration: legs yellow, carapace and abdomen scuta brown,
abdominal interscutal areas creamy white, booklung covers reddish, pedipalps reddish
brown. Habitus as in Fig. 4A, C, E. Carapace (Fig. 4B, F): wide oval in dorsal view;
sides with longitudinal streaks; median area smooth with some setae at lateral edges.
Eyes (Fig. 4B, G): ALE largest, PLE smallest; posterior eye row recurved viewed from
above, procurved from front; ALE almost touching, ALE–PLE separated by less than
ALE radius, PME touching throughout most of their length, PLE–PME separated
by less than PME radius. Clypeus height about 1.7 times ALE diameter (Fig. 4G). Taxonomy nov., female (IZCAS Ar-25106) A, C, E habitus, dorsal, ventral and
lateral views B, D, F prosoma, dorsal, ventral and lateral views. Scale bars: 0.4 mm. Figure 6. Opopaea zhigangi sp. nov., female (IZCAS Ar-25106) A, C, E habitus, dorsal, ventral and
lateral views B, D, F prosoma, dorsal, ventral and lateral views. Scale bars: 0.4 mm. panded cymbiobulbus (the ratio of width to length about 0.47) and the female has a
relatively larger postgynal depression (Platnick and Dupérré 2009: figs 55–66). i
Description. Male (holotype). Measurements: TL: 1.54; CL: 0.60; CW: 0.58;
AL: 0.98; AW: 0.70; ALE: 0.08; PME: 0.07; PLE: 0.07; EGW: 0.22; ALE–ALE: 0.02;
ALE–PLE: 0.01; PME–PME: 0; PLE–PME: 0; CBL: 0.31; CBW: 0.11; PTL: 0.41;
FI: 0.21; FML: 0.14. Coloration: legs yellow, carapace and abdomen scuta brown,
abdominal interscutal areas creamy white, booklung covers reddish, pedipalps reddish
brown. Habitus as in Fig. 4A, C, E. Carapace (Fig. 4B, F): wide oval in dorsal view;
sides with longitudinal streaks; median area smooth with some setae at lateral edges. Eyes (Fig. 4B, G): ALE largest, PLE smallest; posterior eye row recurved viewed from
above, procurved from front; ALE almost touching, ALE–PLE separated by less than
ALE radius, PME touching throughout most of their length, PLE–PME separated
by less than PME radius. Clypeus height about 1.7 times ALE diameter (Fig. 4G). Yanfeng Tong et al. / ZooKeys 917: 51–61 (2020) 60 Yanfeng Tong et al. / ZooKeys 917: 51–61 (2020)
60
Figure 7. Female copulatory organ A–C Opopaea kanpetlet sp. nov. (IZCAS Ar-25099) D–F Opopaea
zhigangi sp. nov. (IZCAS Ar-25106) A, B, D, E ventral view C, F dorsal view. Abbreviations: apo = ap
odeme; asr = anterior scutal ridge; ga = globular appendix; nlp = nail-like process; pd = postgynal depres
sion; pls = paddle-like sclerite; psr = posterior scutal ridge. Scale bars: 0.1 mm. Figure 7. Female copulatory organ A–C Opopaea kanpetlet sp. nov. (IZCAS Ar-25099) D–F Opopaea
zhigangi sp. nov. (IZCAS Ar-25106) A, B, D, E ventral view C, F dorsal view. Abbreviations: apo = ap
odeme; asr = anterior scutal ridge; ga = globular appendix; nlp = nail-like process; pd = postgynal depres
sion; pls = paddle-like sclerite; psr = posterior scutal ridge. Scale bars: 0.1 mm. Figure 7. Female copulatory organ A–C Opopaea kanpetlet sp. nov. (IZCAS Ar-25099) D–F Opopaea
zhigangi sp. nov. Taxonomy (IZCAS Ar-25106) A, B, D, E ventral view C, F dorsal view. Abbreviations: apo = ap
odeme; asr = anterior scutal ridge; ga = globular appendix; nlp = nail-like process; pd = postgynal depres
sion; pls = paddle-like sclerite; psr = posterior scutal ridge. Scale bars: 0.1 mm. Sternum (Fig. 4D) longer than wide, fused to carapace; surface smooth; radial furrows
present between coxae I–II, II–III, III–IV, with rows of small pits. Endites anteriorly
with small, sharply pointed projection. Abdomen: booklung covers very small, red
dish brown, ovoid, without setae. Pedicel tube short, ribbed, with small, dorsolateral,
triangular extensions, scuto-pedicel region lower than pedicel diameter, with arched
scutal ridges, with curved anterior scutal ridge (Fig. 4H, I). Palp (Figs 4J–L, 5): femur
slightly shorter than half length of patella and submedially inserted to patella; patella
very large; tibia small, rounded; cymbiobulbus shorter than the patella; fenestra large
slit-like, located at nearly tip of cymbiobulbus. Tip of embolus broad triangle. Sternum (Fig. 4D) longer than wide, fused to carapace; surface smooth; radial furrows
present between coxae I–II, II–III, III–IV, with rows of small pits. Endites anteriorly
with small, sharply pointed projection. Abdomen: booklung covers very small, red
dish brown, ovoid, without setae. Pedicel tube short, ribbed, with small, dorsolateral,
triangular extensions, scuto-pedicel region lower than pedicel diameter, with arched
scutal ridges, with curved anterior scutal ridge (Fig. 4H, I). Palp (Figs 4J–L, 5): femur
slightly shorter than half length of patella and submedially inserted to patella; patella
very large; tibia small, rounded; cymbiobulbus shorter than the patella; fenestra large
slit-like, located at nearly tip of cymbiobulbus. Tip of embolus broad triangle. New Opopaea (Araneae, Oonopidae) from Myanmar 61 Female (n = 1). As in male except as noted. Measurements: TL: 1.84; CL: 0.65;
CW: 0.58; AL: 1.18; AW: 0.87; ALE: 0.09; PME: 0.07; PLE: 0.06; EGW: 0.21; ALE-
ALE: 0.02; ALE–PLE: 0.01; PME–PME: 0; PLE–PME: 0. Habitus as in Fig. 6A, C,
E. Endites without projections. Copulatory organ (Fig. 7D–F): postgynal depression
(pd) very small; dorsally with nail-like process (nlp) connected to paddle-like sclerite
(pls) bearing thick, straight arms. Distribution. Known only from the type locality. Acknowledgments The manuscript benefitted greatly from comments by Dimitar Dimitrov, Arnaud Hen
rard and one anonymous referee. This study was supported by the National Natural
Science Foundation of China (31750002, 31972867, 31530067) to Yanfeng Tong,
and the Southeast Asia Biodiversity Research Institute, Chinese Academy of Sciences
(2015CASEABRI005, Y4ZK111B01) to Shuqiang Li. References Andriamalala D, Hormiga G (2013) Systematics of the goblin spider genus Opopaea (Araneae,
Oonopidae) in Magagascar. Bulletin of the American Museum of Natural History 380:
1–156. https://doi.org/10.1206/828.1 Baehr BC, Harvey MS, Smith HM, Ott R (2013) The goblin spider genus Opopaea in Aus
tralia and the Pacific islands (Araneae: Oonopidae). Memoirs of the Queensland Museum,
Nature 58: 107–338. Brignoli PM (1978) Ergebnisse der Bhutan-Expedition 1972 des Naturhistorischen Museums
in Basel. Araneae: Fam. Oonopidae, Agelenidae, Hahniidae und Mimetidae. Entomolog
ica Basiliensis 3: 31–56. Platnick NI, Dupérré N (2009) The goblin spider genera Opopaea and Epectris (Araneae,
Oonopidae) in the New World. American Museum Novitates 3649: 1–43. https://doi. org/10.1206/664.1 Tong Y, Li S (2013) Six new species of oonopid spiders from Champasak, Laos (Araneae,
Oonopidae). Zootaxa3709: 71–88. https://doi.org/10.11646/zootaxa.3709.1.3 Tong Y, Chen H, Liu S, Li S (2018) A new genus of oonopid spiders from Myanmar (Araneae,
Oonopidae). ZooKeys794: 31–43. https://doi.org/10.3897/zookeys.794.29156 WSC (2020) World Spider Catalog. Version 21.0 Natural History Museum Bern. World Spi
der Catalog. http://doi.org/10.24436/2 [Accessed on: 2020.2.03]
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Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 ARTIGO ORIGINAL ARTIGO ORIGINAL doi: 10.7213/estud.biol.7339
Licenciado sob uma Licença Creative Commons
ISSN 0102-2067 doi: 10.7213/estud.biol.7339
ISSN 0102-2067 Licenciado sob uma Licença Creative Commons Rodrigo de Andrade Kersten[a], Luiz Antonio Acra[b] [a] Doutor em Engenharia Florestal,
Conservação da Natureza pela
Universidade Federal do Paraná
(UFPR), biólogo, docente no Centro
de Ciências Biológicas e da Saúde da
Pontifícia Universidade Católica do
Paraná (PUCPR), Curitiba, PR - Brasil,
e-mail: r.kersten@pucpr.br
[b] Mestrando em Botânica na Universidade
Federal do Paraná (UFPR), docente no
Centro de Ciências Biológicas e da Saúde
da Pontifícia Universidade Católica do
Paraná (PUCPR), Curitiba, PR - Brasil,
e-mail: luiz.acra@pucpr.br Resumo Procurando resgatar a memória de um dos grandes Botânicos do Paraná, este artigo discorre sobre a vida e a obra de Ralph
João George Hertel, professor das Universidades Católica e Federal do Paraná. Formado na primeira turma do curso de
História Natural do estado, Hertel é responsável por artigos até hoje influentes. Dentre algumas de suas principais publica
ções, podem ser citados o artigo de 1949, primeiro no Brasil a considerar ecologia de epífitas vasculares e um dos primeiros
no mundo. Merece destaque, também, a série Contribuições para a fitologia teórica, além dos trabalhos sobre estrutura e
filogenia de Araucaria angustifolia, até hoje largamente citados. [P] Palavras-chave: Botânica. Paraná. Biografia. História da ecologia. Ralph João George Hertel [ ]
Ralph João George Hertel Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279
Recebido: 05/07/2012
Received: 07/05/2012
Aprovado: 24/08/2012
Approved: 08/24/2012
Rodrigo de Andrade Kersten[a], Luiz Antonio Acra[b]
Resumo
Procurando resgatar a memória de um dos grandes Botânicos do Paraná, este artigo discorre sobre a vida e a obra de Ralph
João George Hertel, professor das Universidades Católica e Federal do Paraná. Formado na primeira turma do curso de
História Natural do estado, Hertel é responsável por artigos até hoje influentes. Dentre algumas de suas principais publica
ções, podem ser citados o artigo de 1949, primeiro no Brasil a considerar ecologia de epífitas vasculares e um dos primeiros
no mundo. Merece destaque, também, a série Contribuições para a fitologia teórica, além dos trabalhos sobre estrutura e
filogenia de Araucaria angustifolia, até hoje largamente citados. [P]
Palavras-chave: Botânica. Paraná. Biografia. História da ecologia. [B]
Abstract
In an attempt to bring to light the works of one of the greatest botanists of Parana, this paper reports the life of Ralph João
George Hertel, professor of both Catholic and Federal University of Parana. Hertel graduated on the first Natural History class
from this state and is responsible for some still influent papers. Among his numerous contributions, his 1949 paper was the first
in Brazil and one of the first in the world to describe the ecology of vascular epiphytes. The series of paper entitled “Contribuições
para a fitologia teórica” (contributions to the theoretical study of plants) and the series of studies considering morphology and
phylogeny of Araucaria angustifolia are also important and, despite their age, are still largely cited. [K]
Keywords: Botany. Parana. Biography. History of ecology. [a] Doutor em Engenharia Florestal,
Conservação da Natureza pela
Universidade Federal do Paraná
(UFPR), biólogo, docente no Centro
de Ciências Biológicas e da Saúde da
Pontifícia Universidade Católica do
Paraná (PUCPR), Curitiba, PR - Brasil,
e-mail: r.kersten@pucpr.br
[b] Mestrando em Botânica na Universidade
Federal do Paraná (UFPR), docente no
Centro de Ciências Biológicas e da Saúde
da Pontifícia Universidade Católica do
Paraná (PUCPR), Curitiba, PR - Brasil,
e-mail: luiz.acra@pucpr.br Rodrigo de Andrade Kersten[a], Luiz Antonio Acra[b] Rodrigo de Andrade Kersten[a], Luiz Antonio Acra[b] [a] Doutor em Engenharia Florestal,
Conservação da Natureza pela
Universidade Federal do Paraná
(UFPR), biólogo, docente no Centro
de Ciências Biológicas e da Saúde da
Pontifícia Universidade Católica do
Paraná (PUCPR), Curitiba, PR - Brasil,
e-mail: r.kersten@pucpr.br
[b] Mestrando em Botânica na Universidade
Federal do Paraná (UFPR), docente no
Centro de Ciências Biológicas e da Saúde
da Pontifícia Universidade Católica do
Paraná (PUCPR), Curitiba, PR - Brasil,
e-mail: luiz.acra@pucpr.br Abstract In an attempt to bring to light the works of one of the greatest botanists of Parana, this paper reports the life of Ralph João
George Hertel, professor of both Catholic and Federal University of Parana. Hertel graduated on the first Natural History class
from this state and is responsible for some still influent papers. Among his numerous contributions, his 1949 paper was the first
in Brazil and one of the first in the world to describe the ecology of vascular epiphytes. The series of paper entitled “Contribuições
para a fitologia teórica” (contributions to the theoretical study of plants) and the series of studies considering morphology and
phylogeny of Araucaria angustifolia are also important and, despite their age, are still largely cited. [K] Estud Biol
Ambiente Divers 2012 jul /dez
34(83) 269-279
Recebido: 05/07/2012
Received: 07/05/2012
Aprovado: 24/08/2012
Approved: 08/24/2012
Keywords: Botany. Parana. Biography. History of ecology. Keywords: Botany. Parana. Biography. History of ecology. 270 Kersten, R. A., Acra, L. A. Ralph João George Hertel (1923-1985, Figura 1),
nascido em 12 de junho de 1923 na cidade de Cu
ritiba, foi um dos grandes naturalistas de sua época. Em 1943, ingressou na primeira turma do curso de
História Natural da Faculdade de Filosofia, Ciências
e Letras do Paraná, primeiro do Estado e terceiro do
país. Três anos mais tarde, em 1945, foi diplomado
bacharel com outro grande expoente das Ciências
Biológicas do Paraná e do Brasil, Rudolf Bruno Lange. O curso contava com poucas disciplinas, abrangendo
as principais áreas das ciências ambientais. À épo
ca foram seus professores: Homero de Melo Braga
(Biologia Geral), Padre Jesus Moure (Zoologia), Carlos
Stellfeld (Botânica), Ludwig Hans Weber (Mineralogia
e Petrografia) e Joaquim Monteiro Martins Franco
(Geologia e Paleontologia), que se revezaram na con
dução das disciplinas (Westphalen, 1988). Figura 1- Prof. Ralph Hertel em consulta a seu livro de
anotações
Fonte: Museu de Zoologia, PUCPR, 1969. Ainda na década de 1940, Hertel foi nomeado as
sistente da seção de Botânica, assumindo como pro
fessor efetivo em 1949. Na década de 1950, foi um
dos fundadores da Sociedade Paranaense de Ciências
Naturais (SPCN), de cuja publicação científica, a
Dusenia, ele seria o primeiro editor (Ardigó, 2011). Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 Abstract Doutor em Ciências e livre docente em Botânica,
Hertel foi colaborador do Museu Paranaense, pro
fessor efetivo de Botânica da Faculdade de Filosofia
da Universidade Católica do Paraná, professor da
Universidade Federal do Paraná, chefe da Divisão de
Botânica do Instituto de Defesa do Patrimônio Natural
do Estado do Paraná e presidente da Sociedade
Paranaense de Ciências Naturais além de funda
dor do departamento de Botânica da Universidade
Federal do Paraná. Ao longo de sua carreira, visitou
várias áreas da Botânica, estudando protistas (Hertel,
1957), fungos (Hertel, 1954a, 1954b, 1955, 1956),
pteridófitas (Hertel, 1947), ecologia (Hertel, 1949,
1950, 1952), biogeografia (Hertel, 1959b, 1969), até
finalmente especializar-se em morfologia e evolução
dos órgãos reprodutivos das plantas (Hertel, 1958,
1959, 1960, 1962/63, 1963, 1968, 1974a, 1974b,
1976a, 1976b, 1980, 1984a, 1984b). Morreu aos 62
anos, em 10 de maio de 1985. Figura 1- Prof. Ralph Hertel em consulta a seu livro de
anotações
Fonte: Museu de Zoologia, PUCPR, 1969. desconhecido. Possivelmente, o manuscrito, já pron
to, não foi submetido, pois os volumes IV e V (Hertel,
1960, 1962) aparecem publicados. No ano de 1949, visando ingressar como profes
sor efetivo da instituição, escreveu sua tese de livre
-docência, publicada em 1950 nos arquivos do Museu
Paranaense. Esse trabalho, intitulado “Contribuição
à ecologia da flora epífita da Serra do Mar (vertente
oeste) do Paraná”, foi o primeiro no mundo a consi
derar ecologia de epífitas vasculares, além de um dos
poucos trabalhos no geral a incluir “ecologia” no títu
lo antes de 1950. O termo ecologia (oekologie) foi empregado pela
primeira vez em 1866, na obra Generelle Morphologie
der Organismen, do renomado biólogo Ernst Haeckel. As bases teóricas da ecologia, no entanto, começa
ram a ser estabelecidas muitos anos mais tarde, com
o clássico trabalho de Clements (1916) sobre o con
ceito de comunidade e de clímax. Há certo consenso
entre ecólogos de que, apesar de o termo ecologia
ter sido cunhado em meados do século XIX, apenas
no início do século XX emergiu como disciplina, e Em sua primeira obra publicada, Hertel (1947)
realizou um estudo químico do exsudato liberado pe
los hidatódios de Phlebodium areolatum, comprovan
do serem cristais de carbonato de cálcio que se apre
sentam como pontuações esbranquiçadas na face
adaxial da lâmina. Uma curiosidade sobre suas publi
cações é a série de artigos intitulados Contribuições
para a fitologia teórica, cujo volume III permanece Estud. Biol., Ambiente Divers. Abstract 2012 jul./dez., 34(83), 269-279 271 Ralph João George Hertel apenas na segunda metade deste século ganhou pro
eminência pública (McIntosh, 1985). Este trabalho não pretende senão fornecer uma
primeira idéia sobre as condições ecológicas, das
quais dispõem os vegetais epífitos em determinada
zona do Paraná – a vertente oeste da serra do Mar –. Contém êle as primeiras informações sobre as pro
priedades físicas, químicas e biológicas do subs
trato desta vegetação; além disso, em traços muito
gerais, a composição química das espécies mais im
portantes (Hertel, 1950). Mais impressionante ainda é o trabalho de Ralph
Hertel em 1949, tanto pelo fato de a ciência não es
tar ainda completamente consolidada quanto pela
juventude do autor à época. O texto, idealizado para
ser sua tese de livre-docência (espécie de concurso
público), foi defendida apenas quatro anos após a ob
tenção do diploma superior, e em seguida, publicada
como seu segundo artigo (Hertel, 1950). Nele, Hertel
estudou a distribuição espacial de epífitos sobre dife
rentes espécies de árvores (Figuras 2 e 3), observan
do diversos parâmetros ambientais, tais como lumi
nosidade, pluviosidade, temperatura, e parâmetros
dos “substratos” (forófitos), como inclinação do tron
co, rugosidade e composição química da casca. Além
disso, Hertel estudou interações entre as epífitas, de
finindo o que chamou de “plantas-guia”, Nematanthus
wetsteinii (Figura 3) e Sinningia douglasii. A listagem apresentada a seguir foi retirada desta
publicação e inclui todas as espécies citadas ao longo
do texto como ocorrendo na região. A área, chama
da por ele de “vertente oeste da Serra do Mar” nas
margens do “Rio do Meio”, corresponde atualmente
ao município de Quatro Barras, nas proximidades
da Serra da Graciosa (25º 20’S, 48º 56’W). No artigo
são apresentadas tanto espécies por ele observadas
quanto espécies tombadas no Herbário [sic.] do “Snr. Gerth Hatschbach e do Autor típicas da região”. A lista original apresenta alguns problemas, como
nomes com grafia incorreta, falta de autores, citação de
nomes não válidos e espécies não reconhecidas para o
Brasil. Essas situações refletem a dificuldade, à época,
de determinação específica em razão da falta de lite
ratura especializada, material de referência, e mesmo
de taxonomistas, pois fica claro o auxílio de grandes
cientistas nacionais, dentre os quais aparecem citados
nos agradecimentos: F. C. Hoehne, A, C, Brade, Graziela
Barrozo, Fritz Lange de Morretes e Rudolf B, Lange. Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 Abstract Figura 2 - Algumas das ilustrações de Hertel sobre a distri
buição das epífitas sobre exemplar de Vitex mon
tevidensis (esq.) e de Ilex paraguariensis (dir.)
Fonte: Hertel, 1949. Figura 3 - Uma das imagens apresentadas na tese de Hertel,
Habitáculo 1 (Sobre Clethra?). À esquerda (do ob
servador), ramos de Hypocyrta wettsteinii; à di
reita, Bromeliaceae; no centro, Araceae. Sôbre as
raízes de Hypocyrta (em claro) é visível a cobertura
de Bryophyta, onde foi tomada a temperatura [sic]. Fonte: Hertel, 1949. Figura 2 - Algumas das ilustrações de Hertel sobre a distri
buição das epífitas sobre exemplar de Vitex mon
tevidensis (esq.) e de Ilex paraguariensis (dir.)
Fonte: Hertel, 1949. Fonte: Hertel, 1949. Seguem os primeiros parágrafos do texto como
publicados pelo autor: Figura 3 - Uma das imagens apresentadas na tese de Hertel,
Habitáculo 1 (Sobre Clethra?). À esquerda (do ob
servador), ramos de Hypocyrta wettsteinii; à di
reita, Bromeliaceae; no centro, Araceae. Sôbre as
raízes de Hypocyrta (em claro) é visível a cobertura
de Bryophyta, onde foi tomada a temperatura [sic]. Fonte: Hertel, 1949. Com o presente trabalho apresentamos uma pri
meira contribuição ao estudo ecológico da flora
epífita do Paraná e, quiçá, do País, pelo menos não
chegou às nossas vistas nenhum trabalho nacional
que se relacione diretamente com o assunto, não
obstante insistente procura que vimos realizando
há vários anos. Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 272 Kersten, R. A., Acra, L. A. estão integradas ao SpeciesLink (BHCB, FLOR, FUEL,
JBRJ_RB, MOBOT, NMNH-Botany, NYBG_BR, SP, UB,
UFP). Em alguns casos, foi necessária a consulta à
descrição original das espécies para a correta deter
minação do nome válido quando a espécie, segundo
a Flora do Brasil, não ocorria no Paraná. As espécies
foram redeterminadas, a partir dos desenhos da pu
blicação, de coletas depositadas no MBM e consultas
a especialistas. Em alguns casos, foram considerados
erros de determinação, sendo utilizado o nome de es
pécies afins, segundo a descrição original, e comuns
na região. Foram citadas ao longo do texto 95 espécies
distribuídas em 60 gêneros e 20 famílias, incluindo ao
menos 18 espécies ainda não citadas em trabalhos
publicados com epífita no Brasil e diversas espécies
não mais observadas na região. O artigo representa um marco pouco reconhecido
na ciência brasileira, merecendo a atenção aqui deferi
da. Para a atualização dessa lista, foi necessário longo
trabalho de resgate taxonômico. Abstract No Quadro 1 foram
incluídos os nomes como aparecem escritos no texto,
sendo consideradas todas as espécies citadas ao longo
do artigo. As espécies tiveram seu nome atualizado
de acordo com pesquisa nas bases de dados da Flora
do Brasil, Tropicos, IPNI e World Checklist of Selected
Planta Families, às vezes apenas após consulta aos
quatro bancos o nome válido foi determinado. Foram
também consultadas as coletas do autor tombadas
no Museu Botânico Municipal de Curitiba (MBM), no
Herbário da Universidade Federal do Paraná (UPCB) e
da Pontifícia Universidade Católica do Paraná (HUCP),
assim como aos demais herbários cujas coleções Quadro 1 - Relação das espécies epifíticas citadas no texto de Hertel (1949) organizadas segundo o nome atualizado,
indicada a página do texto original em que é citada
Nome atualizado
FAMÍLIA
Espécie
Citação original
FAMÍLIA
Espécie
Página
APOCYNACEAE
Mandevilla atroviolacea (Stadelm.) Woodson
Mandevilla immaculata Woodson
APOCYNACEAE
Mandevilla atroviolacea (Stadeln.) Woods. Mandevilla immaculata Woods. 11
11
ARALIACEAE
Hydrocotyle quinqueloba Ruiz et Pav.1
APIACEAE
Hydrocotyle quinqueloba Ruiz et Pav. 15
ARACEAE
Philodendron loefgrenii Engl. ARACEAE
Monstera pertusa (L.) de Vriese ?2
12
ASPLENIACEAE
Asplenium auritum Sw.3
POLYPODIACEAE
Polypodium auritum Sw. 21
BOMBACACEAE
Spirotheca rivieri (Decne.) Ulbr. BOMBACACEAE
Ceiba Rivieri Schum. 11
BEGONIACEAE
Begonia fruticosa A. DC. BEGONIACEAE
Begonia fruticosa A. DC. 14
BROMELIACEAE
Aechmea ornata Baker
Nidularium innocentii Lem. Tillandsia recurvata (L.) L. Tillandsia usneoides L. Vriesea incurvata Gaudich. BROMELIACEAE
Aechmea hystrix ? Nidularia innocenti ? Tillandsia bandensis ? Tillandsia usneoides L. Vriesia incurvata ? 12
12
12
12
12
(Continua) 1 Espécie não citada para o Paraná nem nunca citada como epífita, mas continuamente mencionada pelo autor como hemiepífita. 2 Identificada via foto, estampa 1a. 3 Non Polypodium auritum (Hook.) E. J. Lowe syn. Thelypteris aurita (Hook.) Ching, planta originária do Nepal/China. Asplenium auritum
é a única pteridófita epífita com este epíteto no Brasil e tem Swartz como autor. Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 Ralph João George Hertel 273 Quadro 1 - Relação das espécies epifíticas citadas no texto de Hertel (1949) organizadas segundo o nome atualizado,
indicada a página do texto original em que é citada
Nome atualizado
FAMÍLIA
Espécie
Citação original
FAMÍLIA
Espécie
Página
CACTACEAE
Rhipsalis spp. CACTACEAE
Rhipsalis spp
14
COMMELINACEAE
Indet.4
COMMELINACEAE
Indet.4
14
DRYOPTERIDACEAE
Rumohra adiantiformis (G. Forst) Ching
POLYPODIACEAE
Polystichum adiantiforme (Forst.) J. Sm. 16
GESNERIACEAE
Nematanthus gregarius D.L. Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 4 Possivelmente, refere-se à Tradescantia fluminensis, única espécie da família registrada como epífito na região atualmente.
5 Hypocyrta radicans Klotzsch & Hanst. (1864) nom. syn. Nematanthus radicans (Klotzsch & Hanst.) H.E. Moore (1973) nom. illeg., non
Nematanthus radicans C. Presl (1845) nomen novum Nematanthus gregarius D.L. Denham (1974)
6 T. tenerum inexistente. T. tenellum, syn T. capillaceum.
7 Lycopodium comans H. Christ (1900) nom. illeg., n.v. Lycopodium comans Hook. f. (1844) [nom. syn. Huperzia comans (Herter ex Nessel)
B. Øllg. & P.G. Windisch (1987)].
8
d h
(
d h
)
d
A
ô
l
f
d d
à
l é 4 Possivelmente, refere-se à Tradescantia fluminensis, única espécie da família registrada como epífito na região atualmente.
5 Hypocyrta radicans Klotzsch & Hanst. (1864) nom. syn. Nematanthus radicans (Klotzsch & Hanst.) H.E. Moore (1973) nom. illeg., non
Nematanthus radicans C. Presl (1845) nomen novum Nematanthus gregarius D.L. Denham (1974)
6 T. tenerum inexistente. T. tenellum, syn T. capillaceum.
7 Lycopodium comans H. Christ (1900) nom. illeg., n.v. Lycopodium comans Hook. f. (1844) [nom. syn. Huperzia comans (Herter ex Nessel)
B. Øllg. & P.G. Windisch (1987)].
8 L. dichotomum (syn. Huperzia dichotoma) citada apenas para a Amazônia, possivelmente confundida com H. comans, à qual é muito
similar.
9 Repert. Spec. Nov. Regni Veg. 39, 70, t. 194b (1935). Abstract Denham
Nemathanthus wettsteinii (Fritsch) H.E.Moore
Sinningia douglasii (Lindl.) Chautems
GESNERIACEAE
Hypocyrta radicans Hanst.5
Hypocyrta Wettsteinii Fritsch. Corytholoma sp. 12
12
14-15
HYMENOPHYLLACEAE
Hymenophyllum caudiculatum Mart. Hymenophyllum cf. ciliatum Sw. Hymenophyllum lineare Sw. Trichomanes emarginatum Poir. Trichomanes hymenoides Hedw. Trichomanes radicans Sw. Trichomanes capillaceum L6
Não citada para o Brasil
HYMENOPHYLLACEAE
Hymenophyllum caudiculatum Mart. Hymenophyllum ciliatum ? Hymenophyllum lineare Sw. var brasiliense Ros. Trichomanes emarginatum Pr. Trichomanes hymenoides Hedw. Trichomanes radicans Spr. Trichomanes tenerum Spr. Hymenophyllum interruptum Kze. 11
11
11
11
11
11
11
11
LOMARIPSIDACEAE
Elaphoglossum ornatum (Mett.) Christ. POLYPODIACEAE
Elaphoglossum ornatum (Mett.) Chr. 11
LYCOPODIACEAE
Huperzia comans (Nessel) B. Ollgard & P.G. Windisch
Huperzia comans (Nessel) B. Ollgard & P.G. Windisch **
Huperzia fontinaloides (Sprig.) Trevis. Huperzia heterocarpon (Fée) Holub9
Huperzia taxifolia (Sw.) Trevis. LYCOPODIACEAE
Lycopodium comans Chr.7
Lycopodium dichotomum Jacq.8
Lycopodium fontinaloides Spring. Lycopodium portoanus (Nessel)
Lycopodium taxifolium Sw. var nitens Poepp. 11
11
11
11
11
11
MELASTOMATACEAE
Pleiochiton ebracteatum Triana
MELASTOMATACEAE
Pleiochiton ebracteatum Trian. 12
ONAGRACEAE
Fuchsia regia (Vauld) Munz. OENOTHERACEAE
Fuchsia regia (Vaud.) Mnz. 12
(Continua) 4 Possivelmente, refere-se à Tradescantia fluminensis, única espécie da família registrada como epífito na região atualmente. 5 Hypocyrta radicans Klotzsch & Hanst. (1864) nom. syn. Nematanthus radicans (Klotzsch & Hanst.) H.E. Moore (1973) nom. illeg., non
Nematanthus radicans C. Presl (1845) nomen novum Nematanthus gregarius D.L. Denham (1974)
6 T. tenerum inexistente. T. tenellum, syn T. capillaceum. 7 Lycopodium comans H. Christ (1900) nom. illeg., n.v. Lycopodium comans Hook. f. (1844) [nom. syn. Huperzia comans (Herter ex Nessel)
B. Øllg. & P.G. Windisch (1987)]. 8 L. dichotomum (syn. Huperzia dichotoma) citada apenas para a Amazônia, possivelmente confundida com H. comans, à qual é muito
similar. 9 Repert. Spec. Nov. Regni Veg. 39, 70, t. 194b (1935). 9 Repert. Spec. Nov. Regni Veg. 39, 70, t. 194b (1935). Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 274 Kersten, R. A., Acra, L. A. Quadro 1 - Relação das espécies epifíticas citadas no texto de Hertel (1949) organizadas segundo o nome atualizado,
indicada a página do texto original em que é citada
Nome atualizado
FAMÍLIA
Espécie
Citação original
FAMÍLIA
Espécie
Página
ORCHIDACEAE
ORCHIDACEAE
Acianthera hygrophila (Barb.Rodr.) Pridgeon & M.W.Chase
Pleurothallis hygrophila Rodr. 13
Acianthera leptotifolia (Barb.Rodr.) Pridgeon & M.W.Chase
Pleurothallis leptotifolia Rodr. 13
Acianthera luteola (Lindl.) Pridgeon & M.W.Chase
Pleurothallis caespitosa Rodr. 13
Acianthera sonderiana (Rchb.f.) Pridgeon & M.W.Chase
Pleurothallis Sonderana Rchb. f. 10 Maxillaria Regnelii inexistente, o autor referia-se, possivelmente, a M. regeliana, coletadas apenas duas vezes (uma em SC e outra no RJ).
Na descrição é citada como espécie afim de M. picta. 11 Maxillaria echinatum inexistente. Os nomes semelhantes M. echinophyta Barb. Rodr. [nom. syn. Christensonella echinophyta (Barb.
Rodr.) Szlach., Mytnik, Górniak & Smiszek]; Oncidium echinatum (Barb.Rodr.) Cogn. (1905) nom. illeg. [nom. syn. Gomesa echinata
(Barb.Rodr.) M.W.Chase & N.H.Williams (1853)], ou Oncidium echinatum Kunth (1815) [nom. syn. Baptistonia echinata Barb.Rodr.
(1853)] não são citados para o Paraná. A maior possibilidade é Maxillaria echinochila Kraenzl (syn. M. madida e M. subulata), também
citada como M. acicularis e M. hatschbachii, todas sinônimos. 12 Epidendrum imbricatum Lindl. (1831) nom. illeg., non Epidendrum imbricatum Lam. (1793). Nomen novum Epidendrum ramosum var.
imbricatum Ames, F.T. Hubb. & C. Schweinf. (1934). Abstract 13
Alatiglossum uniflorum (Booth.) Baptista
Oncidium uniflorum Booth. 13
Anathallis dryadum (Schltr.) F.Barros
Pleurothallis dryadum Schltrt.. 13
Anathallis piratiningana (Hoehne) F.Barros
Pleurothallis barboselloides
13
Barbosella miersii (Lindl) Schltr. Barbosella Miersii (Reichb. f.) Schltr. 12
Bifrenaria aureafulva Lindl. Bifrenaria aureo-fulva Lindl. 12
Brasilidium concolor (Hook.) F.Barros & V.T.Rodrigues. Oncidium concolor Hook. 13
Brasilidium concolor (Hook.) F.Barros & V.T.Rodrigues. Oncidium Ottonis
13
Brasilidium crispum (Lodd.) Campacci
Oncidium crispum Lodd
13
Brasilidium praetextum (Rchb.f.) Campacci
Oncidium praetextum Reichb. f. 13
Brasiliorchis picta (Hook.) R.B.Singer, S.Koehler &
Carnevali
Maxillaria picta Hook. 12
Brasiliorchis picta (Hook.) R.B.Singer, S.Koehler &
Carnevali
Maxillaria Regnelii10
12
Christensonella subulata (Lindl.) Szlach., Mytnik,
Górniak & Smiszek
Maxillaria acicularis Herb. 12
Christensonella subulata (Lindl.) Szlach., Mytnik,
Górniak & Smiszek. Maxillaria echinatum11
12
Christensonella subulata (Lindl.) Szlach., Mytnik,
Górniak & Smiszek
Maxillaria hatschbachii Schltr. 12
Coppensia hookeri (Rolfe) F.Barros & L.Guimarães
Oncidium Hookerii Rolfe
13
Coppensia loefgrenii (Cogn.) F.Barros & V.T.Rodrigues
Oncidium Loefgrenii Congn. 13
Dryadella aviceps (Rchb.f.) Luer
Masdevallia O’Brieniana Rolfe
12
Dryadella lilliputana (Cogn.) Luer
Masdevallia lilliputana
12
Dryadella zebrina (Porsch) Luer
Masdevallia zebrina Porsch
12
Encyclia oncidioides (Lindl.) Schltr. Epidendrum oncidioides Lindl
12
Epidendrum caldense Barb. Rodr. Epidendrum hatschbachii Schltr. 12
Epidendrum paniculatum Ruiz & Pavón
Epidendrum Noackii Gongn. 12
Epidendrum ramosum Jacq. Epidendrum imbricatum Ldl.12
12
10 Maxillaria Regnelii inexistente, o autor referia-se, possivelmente, a M. regeliana, coletadas apenas duas vezes (uma em SC e outra no RJ). Na descrição é citada como espécie afim de M. picta. 11 Maxillaria echinatum inexistente. Os nomes semelhantes M. echinophyta Barb. Rodr. [nom. syn. Christensonella echinophyta (Barb. Rodr) Szlach Mytnik Górniak & Smiszek]; Oncidium echinatum (Barb Rodr) Cogn (1905) nom illeg [nom syn Gomesa echinata
(Continua) Quadro 1 - Relação das espécies epifíticas citadas
indicada a página do texto original em
Nome atualizado
FAMÍLIA
Espécie
ORCHIDACEAE
Acianthera hygrophila (Barb.Rodr.) Pridgeon & M.W.Chase
Acianthera leptotifolia (Barb.Rodr.) Pridgeon & M.W.Chase
Acianthera luteola (Lindl.) Pridgeon & M.W.Chase
Acianthera sonderiana (Rchb.f.) Pridgeon & M.W.Chase
Alatiglossum uniflorum (Booth.) Baptista
Anathallis dryadum (Schltr.) F.Barros
Anathallis piratiningana (Hoehne) F.Barros
Barbosella miersii (Lindl) Schltr. Bifrenaria aureafulva Lindl. Brasilidium concolor (Hook.) F.Barros & V.T.Rodrigues. Brasilidium concolor (Hook.) F.Barros & V.T.Rodrigues. Brasilidium crispum (Lodd.) Campacci
Brasilidium praetextum (Rchb.f.) Campacci
Brasiliorchis picta (Hook.) R.B.Singer, S.Koehler &
Carnevali
Brasiliorchis picta (Hook.) R.B.Singer, S.Koehler &
Carnevali
Christensonella subulata (Lindl.) Szlach., Mytnik,
Górniak & Smiszek
Christensonella subulata (Lindl.) Szlach., Mytnik,
Górniak & Smiszek. Abstract Christensonella subulata (Lindl.) Szlach., Mytnik,
Górniak & Smiszek
Coppensia hookeri (Rolfe) F.Barros & L.Guimarães
Coppensia loefgrenii (Cogn.) F.Barros & V.T.Rodrigues
Dryadella aviceps (Rchb.f.) Luer
Dryadella lilliputana (Cogn.) Luer
Dryadella zebrina (Porsch) Luer
Encyclia oncidioides (Lindl.) Schltr. Epidendrum caldense Barb. Rodr. Epidendrum paniculatum Ruiz & Pavón
Epidendrum ramosum Jacq. Epidendrum caldense Barb. Rodr. 10 Maxillaria Regnelii inexistente, o autor referia-se, possivelmente, a M. regeliana, coletadas apenas duas vezes (uma em SC e outra no RJ). Na descrição é citada como espécie afim de M. picta. 11 Maxillaria echinatum inexistente. Os nomes semelhantes M. echinophyta Barb. Rodr. [nom. syn. Christensonella echinophyta (Barb. Rodr.) Szlach., Mytnik, Górniak & Smiszek]; Oncidium echinatum (Barb.Rodr.) Cogn. (1905) nom. illeg. [nom. syn. Gomesa echinata
(Barb.Rodr.) M.W.Chase & N.H.Williams (1853)], ou Oncidium echinatum Kunth (1815) [nom. syn. Baptistonia echinata Barb.Rodr. (1853)] não são citados para o Paraná. A maior possibilidade é Maxillaria echinochila Kraenzl (syn. M. madida e M. subulata), também
citada como M. acicularis e M. hatschbachii, todas sinônimos. 12 Epidendrum imbricatum Lindl. (1831) nom. illeg., non Epidendrum imbricatum Lam. (1793). Nomen novum Epidendrum ramosum var. imbricatum Ames, F.T. Hubb. & C. Schweinf. (1934). Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 Ralph João George Hertel 275 Quadro 1 - Relação das espécies epifíticas citadas no texto de Hertel (1949) organizadas segundo o nome atualizado,
indicada a página do texto original em que é citada
Nome atualizado
FAMÍLIA
Espécie
Citação original
FAMÍLIA
Espécie
Página
ORCHIDACEAE
ORCHIDACEAE
Epidendrum ramosum Jacq. Epidendrum ramosum Jacq. 12
Epidendrum secundum Jacq. Epidendrum ellipticum Ldl. 12
Grobya galeata Lindl. Grobya galeata Lindl. 12
Hadrolaelia coccinea (Lindl.) Chiron & V.P.Castro
Sophronites coccinea (Lindl.) Rchb. F. 13
Isabelia pulchella (Kraenzl.) Sebbas & Teuscher
Neolauchea pulchella Kraenzl. 13
Leptotes unicolor Barb. Rodr. Leptotes unicolor Barb. Rodr. 12
Octomeria alpina Barb. Rodr. Octomeria alpina Barb. Rodr. 12
Octomeria sp. Octomeria sp. 14
Ornithophora radicans (Rchb. f.) Garay & Pabst
Sigmatostalix radicans Rchb. f. 13
Pabstiella mirabilis (Schltr.) Brieger & Senghas
Pleurothallis mirabilis Schltr. 13
Pabstiella pleurothalloides (Cogn.) Luer
Pleurothallis Edwallii Dusen et Schltr. 13
Phymatidium delicatulum Lindl. Phymatidium myrtophilum Rodr. 13
Pleurobotrium crepinianum (Congn.) Hoehne
Pleurobotrium crepinianum (Congn.) Hoehne
13
Pleurobotryum hatschbachii (Schltr.) Hoehne
Pleurobotrium hatschbachii (Schltr.) Hoehne
13
Pleurothallis ipyrangana Schltr. Pleurothallis ipyrangana Schltr. 13
Promenae dusenii Schltr. Promenae Dusenii Schltr. 13
Promenae paranaensis Schltr. Promenae paranaensis Schltr. 13
Promenae xanthina Lindl. Promenae xanthina Lindl. 13
Prosthechea fausta (Rchb.f. ex Cogn.) W.E.Higgins. Epidendrum faustum Ldl. Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 14 Identificada via Estampa. 15 Identificada via Estampa. 13 A única espécie de Microstylis tombada nos herbários de Curitiba é M. sertulifera, que tem como nome válido Malaxis excavata (Lindl.)
Kuntze. 16 P. recurvum Kaulf, inexistente; o mais semelhante é P. recurvatum Kaulf, compatível com a ilustração apresentada.
17 Identificada via Estampa. Figura 4 - Fases ontogenéticas da Araucaria angustifólia.
A - Fase Tirodêndrica (juvenil), B - Fase Fero
dêndrica (transicional), C - Fase Senadêndrica
(convexa), D - Fase Senadêndrica (plana-hori
zontal) (adultos)
Fonte: Hertel, 1980. 16 P. recurvum Kaulf, inexistente; o mais semelhante é P. recurvatum Kaulf, compatível com a ilustração apresentada.
17 Id
ifi
d
i E Abstract 12
Rodrigueziopsis eleutherosepala (Barb. Rodr.) Schltr. Rodrigueziopsis eleutherosepala (Rodr.) Schltr. 13
Specklinia grobyi (Bateman ex Lindl.) F.Barros
Pleurothallis Grobyi Ldl. var triliniata Congn. 13
Specklinia trifida (Lindl.) F.Barros
Pleurothallis bicristata Congn. 13
Stelis fraterna Lindl. Stelis inaequalisepala Schltr. 13
Zygopetalum maxillare Lodd. Zygopetalum maxillare Lodd. 13
Zygostates pustulata (Kraenzl.) Schltr. Ornithocephalus Dusenianus Kraenzl. 13
Zygostates pustulata (Kraenzl.) Schltr. Zygostates pustulata (Kraenzl.) Schltr. 13
Não citada para o Brasil
Microstylis Hieronymi Congn.13
13
PIPERACEAE
Peperomia catharinae Miq.14
Peperomia tetraphylla Hook. & Arn.15
PIPERACEAE
Peperomia sp1. Peperomia sp2
14
18
POLYPODIACEAE
POLYPODIACEAE
Campyloneurum austrobrasilianum (Alston) de la Sota
Polypodium angustifolium Sw. 11
Campyloneurum nitidum (Kaulf.) C. Presl
Polypodium phyllitides L. 11
Microgramma squamulosa (Kaulf.) de la Sota
Polypodium squamulosum Klf. 11
Niphidium crassifolium (L.) Lellinger
Polypodium crassifolium L. 11
13 A única espécie de Microstylis tombada nos herbários de Curitiba é M. sertulifera, que tem como nome válido Malaxis excavata (Lindl.)
Kuntze
(Continua) 276 Kersten, R. A., Acra, L. A. Quadro 1 - Relação das espécies epifíticas citadas no texto de Hertel (1949) organizadas segundo o nome atualizado,
indicada a página do texto original em que é citada
Nome atualizado
FAMÍLIA
Espécie
Citação original
FAMÍLIA
Espécie
Página
POLYPODIACEAE
Pecluma camptophyllaria (Fée) M.G.Price
Pecluma recurvata (Kaulf.) M. G. Price16
Pecluma sicca (Lindm.) M.G. Price **
Pleopeltis astrolepis (Liebm.)E. Fourn. Pleopeltis hirsutissimua Raddi.17
Pleopeltis macrocarpa (Bory ex Willd.) Kaulf. Serpocaulon catharinae (Langsd. & Fisch.) A.R.Sm. POLYPODIACEAE
Polypodium cinerascens Lindl. Polypodium recurvum Klf. Polypodium plumula H. B. W. Pleopeltis astrolepis (Liebm.)E. Fourn. Polypodium lepidopteris (Langsd. et Fisch)
Polypodium lanceolatum L. Polypodium catharinae Langsd. & Fisch
11
11
11
11
11
11
11
PTERIDACEAE
Vittaria lineata (L.) Sm. VITTARIACEAE
Vittaria lineata (L.) Sm. Fonte: Hertel, 1949. (Conclusão) Gymnospermae. No entanto, as suas “sementes” são
providas de um envoltório constituído pela folha fér
til, por muitos denominada “carpelo” [...]. Após esse artigo, Hertel publicou alguns outros so
bre ecologia (Hertel, 1952), Micologia (Hertel, 1954a,
1954b, 1955, 1956) e um léxico botânico em parce
ria com um dos mais renomados botânicos da épo
ca, Alberto José de Sampaio (Sampaio & Hertel, 1949,
1950). Em 1958, com o início da série de artigos intitu
lada Contribuição para a fitologia teórica, Ralph final
mente começa a publicar o que seria sua grande paixão –
morfologia e evolução vegetal – e que levaria à segunda
grande contribuição do autor para a Ciência Botânica. 17 Identificada via Estampa. Figura 6 - Ilustração de diferentes lígulas registradas em
espécies atuais Sua argumentação está baseada de diversas ca
racterísticas morfológicas, dentre elas, as principais
referem-se à semente. Sua casca consiste do ma
crosporófilo associado à região hipótona da folha
(face dorsal) e à lígula (face ventral) que deixam
sempre uma descontinuidade na extremidade ada
xial. É, assim, muito semelhante ao megagametófito
das atuais Licófitas. Fonte: Hertel, 1974b. Fonte: Hertel, 1974b. sempre possui posição abaxial. O desenvolvimento
ontogenético dessa estrutura é também semelhante
ao registrado em Selaginella e em Isoetes, porém di
ferente de outras espécies de espermatófitas (Hertel,
1974, 1980). Outros argumentos envolvem os fenô
menos da germinação, o sistema dilucional do cáu
dice, a constituição e o comportamento funcional da
folha, os antomas feminino e masculino, e a raiz. Ancestral Avascular
Riniófitas
Lepidodendron
Licófitas
Trimerofitófitas
Monilófitas
Cladoxylopsidas
Pteridospermas
Espermatófitas
Figura 5 - Cladrograma das Traqueófitas atuais e extintas. Notar o posicionamento taxonômico de Lepido
dendron próximo às atuais Licófitas. Fonte: Dados da pesquisa. Em sua tese de livre-docência, Hertel (1980)
conclui que o Araucaria angustifolia, é muito seme
lhante, no que tange nos caracteres teleomáticos, às
Lycopodiidae paleozoicas. O que as separa incondi
cionalmente é o aspecto do antoma masculino, espo
rangial e não esporofilar, como é o feminino. Assume
também ser viável admitir-se a origem comum das
Lycopodiales e das Araucariales, talvez a partir
de formas ainda não conhecidas. Nesse sentido, a
Araucaria angustifolia seria uma Lepidosperma, e
não uma Gimnosperma, como usualmente assumido. Figura 5 - Cladrograma das Traqueófitas atuais e extintas. Notar o posicionamento taxonômico de Lepido
dendron próximo às atuais Licófitas. Dentre suas outras publicações, até hoje citadas
em artigos científicos (Fagundes & Mariath, 2010;
Krupek & Ribeiro, 2010; Souza, Oliveira-Oyama &
Muneratto, 2008), merecem destaque pontual o inti
tulado Contribuições para a fitologia teórica, volume II. Algumas concepções na carpologia (Hertel, 1959a) es
tão dentre as mais citadas do autor, além das séries so
bre os Myxomycetes do Brasil (Hertel, 1954a, 1954b,
1955, 1956) e sobre a imbuia (Hertel, 1968, 1974a). Fonte: Dados da pesquisa. Fonte: Dados da pesquisa. É a lígula justamente um dos focos principais des
ta argumentação. Utilizado em diferentes sentidos e
para diferentes estruturas (Figura 6), o termo lígula
refere-se, na araucária, à porção da folha aderida ao
tronco, observada também no esporofilo que recobre
o esporângio. Abstract Base do esporófilo de Isoetes, com esporângio (e) e língula (1). Figura 6 - Ilustração de diferentes lígulas registradas em
espécies atuais Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 Abstract Kaulf compatível com a ilustração apresentada
Figura 4 - Fases ontogenéticas da Araucaria angustifólia. A - Fase Tirodêndrica (juvenil), B - Fase Fero
dêndrica (transicional), C - Fase Senadêndrica
(convexa), D - Fase Senadêndrica (plana-hori
zontal) (adultos)
Fonte: Hertel, 1980. Profundo conhecedor da morfologia vegetal,
Hertel publicou artigos sobre diversas espécies, no
entanto, sua grande paixão foi a Araucaria angusti
fólia, que lhe rendeu três artigos (Hertel, 1973, 1974,
1976b) e uma tese (Hertel, 1980). Nesta, descreve
diversas características importantes da espécie, des
de forma da copa (Figura 4) até estudos filogenéticos
sobre a origem da lígula. Sua controversa contribuição com este trabalho
(Hertel, 1980) foi quanto à classificação taxonômica
da araucária, Em torno da Araucaria angustifolia (Bertoloni)O.Kuntze
impera profunda mal-compreensão quanto à sua
verdadeira posição dentro do sistema classificatório
cientificamente evoluído. É ela situada, por comodi
dade, no grupo das coníferas e, por tanto, pertence às Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 Ralph João George Hertel 277 Baseando seus argumentos tanto em fatores his
tóricos quanto em morfológicos, ele afirmou que a
araucária deveria ser corretamente classificada como
Lepidosperma, ramo derivado dos Lepidondendron
(Figura 5). Historicamente, argumenta que os pri
meiros cientistas, devido à semelhança da “flor” da
araucária com uma pinha e ao fato de seus “frutos”
terem sido confundidos com os “pinhões” do Pinus
pinea, cujas grandes sementes são também consu
midas. Segundo ele, esse juízo científico (sic) inicial
levou à classificação automática dessa espécie como
“pinheiro do Brasil”, fato que gerou consequências
em seu posicionamento taxonômico. Fig. 1. a - Pétalo de Caryophyllaceae e b - Flor de Compositae, exemplificando os conceitos de língula entre Angiospermae. Fig. 2. Micrófilo de Selaginella, com a tenra língula em posição axiliar. Fig. 3. Base do esporófilo de Isoetes, com esporângio (e) e língula (1). Figura 6 - Ilustração de diferentes lígulas registradas em
espécies atuais
Fonte: Hertel, 1974b. Fig. 1. a - Pétalo de Caryophyllaceae e b - Flor de Compositae, exemplificando os conceitos de língula entre Angiospermae. Fig. 2. Micrófilo de Selaginella, com a tenra língula em posição axiliar. Fig. 3. Base do esporófilo de Isoetes, com esporângio (e) e língula (1). Fig. 1. a - Pétalo de Caryophyllaceae e b - Flor de Compositae, exemplificando os conceitos de língula entre Angiospermae. Fig. 2. Micrófilo de Selaginella, com a tenra língula em posição axiliar. Fig. 3. Referências Hertel, R. J. G. (1960). Contribuições para a fitologia teórica
IV: Sobre a estrutura anátomo-morfológica e fisiológi
ca. Humanitas, 5, 73-104. Ardigó, F. (2011). Histórias de uma ciência Regional, cien
tistas e suas instituições no Paraná (1940-1960). São
Paulo: Editora Contexto. doi:10.5962/bhl.title.56234. Hertel, R. J. G. (1962/63). Contribuição para a fitologia te
órica. V. Da germinação nos vegetais. Humanitas, 6,
101-135. Clements, F. E. (1916) Plant succession: An analysis of the de
velopment of vegetation. Washington: Carnegie Institute. Hertel, R. J. G. (1963). Estudos sobre Araucaria angustifo
lia. I – Descrição morfológica do fruto, a germinação. Boletim do Instituto de História Natural - Botânica, 4. Fagundes, N. F., & Mariath, J. E. A. (2010). Morfoanatomia e
ontogenia de fruto em espécies de Bromeliaceae. Acta
Botânica Brasilica, 24, 765-779. Hertel, R. J. G. (1968). Estudos sobre a Phoebe porosa
(Nees) Mez. I. Nomenclatura da imbuia e alguns dos
problemas que encerra. Dusenia, 8, 164-193. Hertel, R. J. G. (1947). Observações sôbre Polypodium areola
tum H. B. K. Arquivos do Museu Paranaense, 6, 299-339. Hertel, R. J. G. (1969). Aspectos Interessantes da Vegetação
do Paraná. In F. El-Khatib (Ed.). História do Paraná. (Vol. 2, pp. 131-124). Curitiba: Grafipar. Hertel, R. J. G. (1949). Contribuição à ecologia de flora epi
fítica da serra do mar (vertente oeste) do Paraná. Tese
Livre Docência, Faculdade de Filosofia, Ciências e
Letras da Universidade Federal do Paraná (UFPR), Cu
ritiba. PMid:18148212. Hertel, R. J. G. (1974a). Estudos sobre Phoebe porosa (Nees)
Mez II. A inflorescência, a flor e o fruto da imbuia. Acta
Biológica Paranaense, 3, 25-54. Hertel, R. J. G. (1950). Contribuição à ecologia de flora epi
fítica da serra do mar (vertente oeste) do Paraná. Ar
quivos do Museu Paranaense, 8, 3-63. Hertel, R. J. G. (1974b). Uma interpretação filogenética da
lígula. Acta Biológica Paranaense, 3, 55-71. Hertel, R. J. G. (1952). Contribuição ao estudo das associa
ções zoofíticas. Dusenia, 3, 463-470. Hertel, R. J. G. (1976a). Estudo sobre Araucaria angus
tifolia II. A constituição do estróbilo. Acta Biológica
Paranaense, 5, 3-25. Hertel, R. J. G. (1954a). Myxomycetes do Brasil I. Lista dos
Myxomycetes assinalados para o Brasil e descrição de
novas espécies do gênero Arcyria Wiggers. Dusenia, 5,
117-124. Hertel, R. J. G. (1976b). Selecta phytoteratológica I. Teratoma
e placentoma em Carica papaya L. Acta Biológica Para
naense, 5, 27-43. Hertel, R. J. G. (1954b). Myxomycetes do Brasil. II. Paradia
cheopsis curitibana Hertel, n.gen. Figura 6 - Ilustração de diferentes lígulas registradas em
espécies atuais Como regra, pode-se estabelecer que
apenas formas heterosporadas são portadoras de lí
gula (no sentido da observada na Araucária), a qual Espera-se, com este texto, contribuir para o resga
te intelectual de tão brilhante figura que durante dé
cadas ajudou a alavancar a ciência paranaense e bra
sileira, e a formar inúmeros outros pesquisadores. Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 Kersten, R. A., Acra, L. A. 278 Sampaio, A. J., & Hertel, R. J. G. (1950). Lexico botânico,
plantas criptogamicas (celulares e vasculares). Publi
cações avulsas do Museu Paranaense, 2, 1-30. Sampaio, A. J., & Hertel, R. J. G. (1949). Lexico botânico, plan
tas criptogamicas (celulares e vasculares). Publicações
avulsas do Museu Paranaense, 1, 1-31. Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 Westphalen, C. M. (1988). Faculdade de Filosofia, Ciências
e Letras da Universidade Federal do Paraná (UFPR)- 50
anos. Curitiba: SBPH. PMid:2450191. Souza, L. A., Oliveira-Oyama, S., & Muneratto, J. C. (2008).
Morphology and anatomy of the developing fruit of
Macfadyena unguis-cati (L.) A. H. Gentry, Bignoniaceae.
Acta Botânica Venezuelan, 31, 1-14. Referências e n.sp. de Lampro
dermaceae. Dusenia, 5, 191-192. Hertel, R. J. G. (1980). Interpretação morfológica da
Araucaria angustifolia. Tese Professor Titular, Univer
sidade Federal do Paraná (UFPR), Curitiba. Hertel, R. J. G. (1955). Myxomycetes do Brasil III. Dois no
vos elementos de Stemonitaceae. Dusenia, 6, 47-48. Hertel, R. J. G. (1982). O fruto de Celtis trifolia e a interpre
tação morfológica do Teloma. Dusenia, 13, 183-197. Hertel, R. J. G. (1956). Taxonomia de Comatricha Preuss
emend. Rost. (Myxophyta). Dusenia, 7, 341-350. Hertel, R. J. G. (1984a). Atualização conceitual I, reabilita
ção de um campo científico, a Bionomia Vegetal. Es
tudos de Biologia, 9, 1-19. Hertel, R. J. G. (1957). Isolamento de micro-organismo Pi
peta simples para este fim. Humanitas, 2, 1-57. Hertel, R. J. G. (1984b). Atualização conceitual II, valoroso
sufixo mal interpretado. Estudos de Biologia, 10, 1-24. Hertel, R. J. G. (1958). Contribuição para a fitologia teórica
I. Alguns conceitos na histofilogênese. Humanitas, 3,
178-201. Krupek, R. A., & Ribeiro, V. (2010) Biometria e germinação de
sementes de Araucaria angustifolia (Bert.) O. Kuntze pro
venientes de um remanescente florestal do município de
Turvo, PR. Revista Ciências Exatas e Naturais, 12, 73-89. Hertel, R. J. G. (1959a). Contribuições para a fitologia teó
rica II. Algumas concepções na carpologia. Humanitas,
4, 11-53. McIntosh, R. (1985). The Background of Ecology. Concept
and Theory. New York: Cambridge University Press. doi:10.1017/CBO9780511608537. Hertel, R. J. G. (1959b). Esboço fito-ecológico do litoral cen
tro do Paraná. Forma et functio, 1, 47-78. Estud. Biol., Ambiente Divers. 2012 jul./dez., 34(83), 269-279 Ralph João George Hertel 279 Souza, L. A., Oliveira-Oyama, S., & Muneratto, J. C. (2008). Morphology and anatomy of the developing fruit of
Macfadyena unguis-cati (L.) A. H. Gentry, Bignoniaceae. Acta Botânica Venezuelan, 31, 1-14. Westphalen, C. M. (1988). Faculdade de Filosofia, Ciências
e Letras da Universidade Federal do Paraná (UFPR)- 50
anos. Curitiba: SBPH. PMid:2450191.
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Cloud system for diabetic data analysis
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III.
DATA ANALYZIS The main purpose for the server application is data
analyzing. By using this application, the clinician could create
a complex image about patient evolution and give advices for
improving the treatment. I.
INTRODUCTION Diabetes is a silent disease that does not give pain or other
symptoms, but is very aggressive and dangerous for the human
body if is not treated properly by the patient. Diabetes appears
when the pancreas no longer produces the necessary amount of
insulin and the Blood Glucose (BG) value increases and
remains on a high level. This state is known as hyperglycemia
and treating it inappropriately over time leads to blindness or
leg amputation. Server
Application Patients Fig. 1. System architecture. Fig. 1. System architecture. All this information is sent to a cloud database by using
REST (Representational State Transfer) [1] web services built
into the cloud. In order to maintain the BG level in a safety area, the
patient should inject periodically a given insulin amount. The
problem is that the patient is always in a dilemma because
he/she doesn’t know how much insulin to intake. If the insulin
amount is to small, then the patient remains into
hyperglycemia, but on the other hand, too much insulin can kill
the patient through hypoglycemia. Is not so easy to find the
correct amount of insulin needed on a given situation. The BG
level depends on many factors (quantity of carbohydrates
intaken, physical activity, stress, hormone cycle, etc.) so the
patient should take into consideration all these factors and
calculate the correct insulin bolus. The server application reads these values and builds web
pages where the user can analyze the data and conclude about
patient evolution. A. Glucose profile The presented system helps the patient to analyze his/her
medical history, the BG curve over time, Hyper/Hypoglycemia
events occurred during a given period and other blood glucose
statistics. Furthermore, the system builds a patient database
which stores all the critical situations with which the patient
has been confronted over time and the successful solutions for
those cases. When a new case appears, then the patient could
ask the database to show the solution for a similar case in order
to receive some help on his/her decision. Fig. 2. Blood Glucose profile chart. Cloud system for diabetic data analysis Lucian Nita
Faculty of Electrical Engineering
"Gheorghe Asachi" Technical University of Iaşi,
RomSoft SRL
Iasi, Romania Cloud database
Mobile
Application
Patients
Clinicians
WEB services
WEB services
Server
Application
Patients
Fig. 1. System architecture. Abstract—The paper presents a web application which helps
the diabetes patients and clinicians to analyze the patient
evolution and improve the diabetes treatment. The system reads
the blood glucose values from cloud where are uploaded by the
patient mobile application. In this way both patient and clinician
can share the patient data and it is no longer necessary the
patient to visit the hospital. Mobile
Application II.
SYSTEM ARCHITECTURE The first component of the system is represented by a
mobile application which runs on patient phone and which
collects patient data: meals, physical activity, blood glucose
values and other information which help the patient to calculate
the insulin injection quantity (Fig. 1). Fig. 2. Blood Glucose profile chart. This helps the clinician to analyze the patient behavior
specific to each day of the week. One very useful tool for analyzing the patient evolution is
the Glucose Profile chart (Fig. 2). This graph collects the blood
glucose values for a long period and then builds a generic day
with these values showing the main trends and statistics for that
period. For a given minute from this generic day the graph
displays on the same vertical axis all BG values having the
same given time which are found on each day of the selected
time interval. D. Hypo/Hypeglycemia events The application identifies automatically the hypo or
hyperglycemia events from the BG curve. For the same generic
day described on paragraph III.A, the page shows the events
number occurred for each hour (Fig. 6). Clicking on an event,
more details are displayed specific to that event: BG curve, the
event time, the extreme BG value and other context parameters
(physical activity, alcohol, etc.). Fig. 3. Displaying the main percentiles of the BG values. Moving the mouse over the graph, the system automatically
calculates the mouse position and the closest BG value for that
position. Then the graph displays a Tool Tip which lists the
main statistic for that selected time. In this way the clinician
could conclude which time zones are most critical for the
patient and give the proper advices in order to minimize the
hypo/hyperglycemia periods. Fig. 6. Hypoglycemic events page. C. Blood Glucose Statistics This page makes a comparison between two time intervals
in order to analyze the patient progress. The page displays the
average BG, standard deviation, the HbA1C [3] values and
comparisons for hypo and hyperglycemia events. Based on these values the system can calculate the mean
and median value and also the main percentiles [2] of these
vectors (Fig. 3). Fig. 5. Blood glucose statistics page. Fig. 3. Displaying the main percentiles of the BG values. Fig. 5. Blood glucose statistics page. B. Data filtering It’s very important for the user to select the exact time
period that he wants to analyze. Therefore, each page of the
application includes a filter where the user selects the time
interval and the data type to be displayed (Fig. 4). Furthermore,
the user can select a specific day from the week. For example,
if the selected day is Monday, then the application selects data
from all Mondays included into the selected time interval. Fig. 6. Hypoglycemic events page. Fig. 4. Data filtering. ACKNOWLEDGMENT This material is based upon the work which is supported by
the European Union through the H2020 “PEPPER” project:
Patient
Empowerment
through
Predictive
Personalized
decision support, http://www.pepper.eu.com. E. Critical cases analyzing The system implements also a database which stores for
each patient his/her critical cases and their solution. A critical
case means that the patient doesn’t know how much insulin to
inject because he/she is in an unusual state given by unusual
actions (big meal, alcohol, cigarettes) cumulated. Fig. 4. Data filtering. A case is considered successful if the BG value remains
into a safety area after the insulin injection. That case is analyzed by the clinician and if is considered successful, then
the case is saved into database to be used in the future as a
solution for that given context. The presented application uses extensively these libraries
and JavaScript language for both major tasks: data transferring
and data displaying. Fig. 8 shows a section of code which
builds rectangles for each hour representing the physical
activity made by the patient. In order to validate the case, the clinician should have a tool
which visualizes the entire context of that case: the BG curve,
physical activity (number of steps for each hour), the number
of carbohydrates intaken on each meal, basal and bolus insulin
and other details like stress, alcohol, cigarettes, hormone cycle,
etc. (Fig. 7). Fig. 8. JavaScript code which displays the patient physical activity. Fig. 8. JavaScript code which displays the patient physical activity. Fig. 7. Case analysis tool. g. 8. JavaScript code which displays the patient physical activity. In order to download data from the cloud database the
application implements AJAX (Asynchronous JavaScript And
XML) [7] calls which transfer the data in the background
without freezing the page during this process. In this way the
application runs smoothly having the same performance as a
desktop application, but it benefits from all the advantages
given by the client-server architecture. IV.
GRAPHICS ON WEB APPLICATIONS It is known that a web application is a better solution
compared with a desktop application from many points of
view: maintenance, installation, user training, visibility, impact,
development costs. But not any task could be resolved by a
web application due to its limits: every page refresh requires
data transferring between server and the web browser, limited
internet bandwidth, few software libraries for data analyzing
and displaying. Members of the PEPPER Group are as follows: project
leader: C. Martin; project management team: D. Brown,
F.Torrent-Fontbona, P. Herrero Vinas, L. Nita, J. Masoud,
J.M.Fern´andez-Real, B. Lopez; co-investigators: A. Aldea,
D.Duce, M. Fernandez-Balsells, P. Gay, P. Georgiou, R. Harrison,B. Innocenti, Y.Leal, N. Oliver, R. Petite, M. Reddy,
C.Roman, J. Shapley, M. Waite and M. Wos. However, during the last years, there is a massive migration
of applications to cloud space. One important reason is given
by the spread of the latest generation phones that incorporate
processing power similar to that of usual computers. In this
context, many software developers had focused on building
open source libraries which help the engineers to develop web
applications with a great user interface. Most of these libraries
are developed on Java Script language and HTML5 standard
[4] which are supported by the modern browsers. The last
version of SVG (Scalable Vector Graphics) [5] combined with
the D3.js (Data-Driven Documents) [6] library gives to the
software engineer a very powerful tool for graphic
development. V.
CONCLUSIONS A web application for data analysis specific to diabetes is
presented. The application uses the latest JavaScript libraries in
graphic representations that allow to build powerful tool for
data displaying and analysis. Fig. 7. Case analysis tool. Fig. 7. Case analysis tool. Data is stored in a cloud database which makes the dialogue
between the patient and the doctor easier and more
comfortable. Another helpful instrument is the table which lists similar
cases with the present case. The clinician could open each case
from the list and analyze the solution for that context and then
decides if the current case should be saved or not into database. REFERENCES [1] RESTful Web Services,
https://www.tutorialspoint.com/restful/restful_introduction.htm [2] Percentile definition, https://en.wikipedia.org/wiki/Percentile [3] Guide to HbA1c, http://www.diabetes.co.uk/what-is-hba1c.html [4] HTML5 Guide, https://developer.mozilla.org/en-
US/docs/Web/Guide/HTML/HTML5 [5] Scalable Vector Graphics,
https://www.w3schools.com/html/html5_svg.asp [6] Data-Driven Documents, https://d3js.org/ [7] AJAX Guide, https://www.w3schools.com/xml/ajax_intro.asp
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En ese sentido, la ausencia del desarrollo de esos temas en los Grundrisse obedece a la
estructura del plan original, el cual cambiaría a principios de los 60s (1862-63),
integrándolos. Esta idea se ve reforzada constatando que El Capital comprende aún más
temas que no eran tratados en profundidad en los Grundrisse o sólo eran mencionados en
estos borradores de fines de los 50s: cooperación, manufactura, gran industria, la
acumulación originaria, etc., como así también los temas que debería incluir un libro sobre
el trabajo asalariado (Rodolsky 2004, 46).
Esta ampliación del plan original implicaría no sólo la integración de los libros
señalados en él, sino la integración de los temas de la competencia y otras formas que nos
acercan a la superficie de la sociedad burguesa. En otras palabras, es esta misma expansión
del capital que pasa del capital en general a formas más desarrolladas, lo que hace que Marx
integre el libro del trabajo asalariado en El Capital, fundamentalmente en el Tomo I.
Mientas que los otros tres libros pendientes quedarían relegados a un próximo desarrollo
(Rodolsky 2004, 50).
Según Rodolsky, el plan de fines de los 50s será un modelo provisorio por medio del
cual Marx desarrolló las “determinaciones puras” (Rodolsky 2004, 68). Para el polaco, “…si
bien se les abandonó en cuanto secciones independientes, incorporando su contenido a la
nueva estructura de la obra, […] la estricta separación originaria de las categorías fue sólo
un medio de abstracción metodológica y que por ello se la pudo abandonar apenas hubo
llevado a cabo la tarea principal: el análisis del “capital en general”” (Rodolsky 2004, 69)
A nuestro parecer, Rodolsky reduce lo simple de las formas como “lo común” en todas
ellas, lo que a veces vuelve confusa su forma de entender el método y las formas del capital
“puro”. Sin embargo, a pesar de esta falta de precisión, entiende que los otros temas ausentes
son la forma desarrollada del capital en general, como son la competencia, la formación de
la tasa de ganancia, etc.; todos fenómenos de la circulación (Rodolsky 2004, 78-79), y que
se tratan en el Tomo III. Esto, si bien se enmarca en la discusión marxológica permite al
autor establecer el carácter desarrollado de El Capital –que se acerca más a las formas
concretas del capital, a sus manifestaciones superficiales- respecto de los Grundrisse -donde
Marx se dedica al desarrollo del capital en general (Rodolsky 2004, 82). En otras palabras,
integra los libros del trabajo asalariado y de la propiedad de la tierra a El Capital, pero, más
importante aún, señala la diferencia específica entre las formas simples y desarrolladas pero
de una manera abstracta. De todas maneras, ni un posible libro del trabajo asalariado o bien
sobre la propiedad de la tierra trataría de formas esenciales, sino que sería el desarrollo de lo
ya embrionario y más o menos desarrollado en El Capital (Rodolsky 2004, 80-81).
Entonces, considerando que El Capital parece ser una forma desarrollada de lo
esbozado en los Grundrisse, el libro del trabajo asalariado se integra al Tomo I porque
263
Gabriel Rivas, Subjetividad revolucionaria y capital. A propósito del “libro” o “teoría” del trabajo asalariado,
Izquierdas, 29:251-274, septiembre 2016
expresa, según Rodolsky, “un eslabón intermedio necesario entre la teoría del valor del tomo
I y la teoría de los precios de la producción, a desarrollar en el tomo III” (Rodolsky 2004,
85). Pero así también, estos cambios en los planes, señala finalmente Rodolsky, demuestran
que “la estricta separación entre las categorías del capital y del trabajo asalariado, tal como
la preveía el plan antiguo, sólo resultaba viable hasta cierto punto” (Rodolsky 2004, 91), a
partir del cual el plan debió ser abandonado.
Sin embargo, nos separamos de Rodolsky a la hora de considerar de forma más precisa
estas extensiones del plan original, el desarrollo de sus formas más concretas, ya que
confunde las partes llamadas “históricas” del capital –y que son parte extensa de, por
ejemplo, la sección sobre el salario, pero así también de la ley general de acumulación
capitalista-, como momentos de carácter ilustrativo y no como las formas desarrolladas del
capital. Según el autor, “los resultados esenciales de la investigación que aparecen en las
secciones mencionadas […] también hubiesen podido exponerse sin necesidad de recurrir a
ese material ilustrativo, cosa que durante años había sido la intención de Marx” (Rodolsky
2004, 90). Frente a este punto, concordamos con Caligaris a la hora de indicar que las partes
que Rodolsky puede identificar como “ilustrativas”, responden al desarrollo de formas más
concretas bajo las cuales debe realizarse la necesidad contenida como potencia en la forma
abstractas. Considerando lo tratado por el autor, la compra y venta de la fuerza de trabajo, la
explotación de la fuerza de trabajo por parte del capitalista, se realiza necesariamente como
lucha de clases (Caligaris 2012.), por lo que aquello que Rodolsky llama puro no puede ser
reducido a su forma simple y solo se comprende como las formas concretas que toma su
desarrollo. Finalmente, creemos que esta limitación en la comprensión del método, reduce el
alcance de la observación de Rodolsky al limitar el problema del libro sobre el trabajo
asalariado a un tema “marxológico”, sin avanzar en las determinaciones de la potencialidad
revolucionaria de la clase obrera, o al menos volviendo su relación una de carácter poco
claro.
e. Más allá de la interpretación de Marx
Finalmente encontramos a Lapides quien, dentro de la tradición de autores marxistas,
parece ser el que ha tratado con mayor rigor el tema del salario en la obra de Marx. En su
trabajo más extenso (Lapides 2002) sostiene que El Capital contiene las determinaciones
esenciales del salario. Haciendo referencia a lo esbozado por Marx en el capítulo XVIII del
Tomo I, donde analiza el trabajo por tiempo -una de las formas concretas que adopta el
salario y que expresa una modificación formal de las “leyes del salario” (Marx 2002, 661),
Lapides plantea que “aquellos escritores que señalan que Marx reservaba el tratamiento de
los salarios para un trabajo separado, aparentemente no han notado esta observación”
(Lapides 2008, 196).
A partir de una revisión de El Capital, Lapides estima que Marx ha creado con éxito
una “teoría” de los salarios. Dinámica, compleja y comprensiva, explica y predice los
diversos fenómenos del salario, así como las principales tendencias de la relación capitaltrabajo (Lapides 2008, 207).
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Gabriel Rivas, Subjetividad revolucionaria y capital. A propósito del “libro” o “teoría” del trabajo asalariado,
Izquierdas, 29:251-274, septiembre 2016
En su opinión, la expresión “teoría del trabajo asalariado” que aparece incluso en El
Capital estaría reservada “para los aspectos puramente fenoménicos del salario. Las
“fricciones” reales, como las llamó, del mercado, tales como el engaño y la coerción que son
inevitables en la economía capitalista pero no son intrínsecos a su dinámica interna”
(Lapides 2008, 208). Esta distinción entre lo fenoménico y lo esencial vacía de necesidad a
estas formas que son subordinadas de manera abstracta a la “dinámica interna” del capital,
perdiendo de vista lo concreto.
Como ya señalamos, Lapides se ubica dentro de los marxistas que rechazan la idea de
un trabajo ausente, “declarando que la teoría presente en El Capital y textos relacionados es,
si es que no literalmente, al menos analíticamente completa” (Lapides 2008, 211). Según
Lapides, esto se debería a un error en la interpretación de la transformación del plan de la
obra, el cual parece haber quedado reducido al primero de los seis libros planteados a fines
de los 50s, de ahí que falte el libro del trabajo asalariado y El Capital quede reducido a una
teoría incompleta (Lapides 2008, 213).
Lapides recorre el debate desde Kautsky –que fue el primero en señalar la divergencia
en los planes, pero sosteniendo el carácter acabado de la “teoría del salario”; Wilbrand –que
es el primero en declarar el carácter incompleto de la obra de Marx, una parte de seis;
Grossman -quien desde un punto un tanto más cercano al nuestro, contesta al segundo desde
“la estructura interna” de El Capital, pero a quien también Lapides critica (Lapides 1997);
Rubel –que revive los argumentos de Wilbrand; terminando con Lebowitz –a quien dedica
un breve artículo (Lapides 2002) y McLellan -ambos sostienen esta idea de un libro faltante
en la misma línea de Wilbrand-Rubel (Lapides 2008, 213-216). Sin embargo, a pesar de
adherir a la idea de que el fundamento del salario está contenido en El Capital cae en la
trampa de la marxología: “Para evaluar estas posiciones, el mejor camino es revisar primero
la evolución misma del plan de Marx para escribir El Capital” (Lapides 2008, 216).
A pesar del carácter marxológico del ejercicio, Lapides concluye –como Rodolsky- que
“la relación capital trabajo está puesta en el corazón del análisis” (Lapides 2008, 221) y
señala que es a principios de los 60s donde Marx desarrolla por primera vez los elementos
fundamentales de su teoría del salario (Lapides 2008, 222), confirmando el abandono del
plan original y su integración a El Capital como forma desarrollada del capital.
Finalmente, avanzando en el impacto político del aporte científico de Marx, Lapides
señala que la teoría del salario y del trabajo asalariado de Marx ha sido un elemento
indispensable en el “arsenal ideológico” de la clase obrera: “pretender que no existe es
intentar desarmar a los obreros negándoles esta parte de su legado” (Lapides 2002, 233). Sin
embargo, nos parece que el análisis marxológico es insuficiente. Sostenemos que el aporte
de Marx no es parte de un “arsenal ideológico” siendo el desarrollo científico del problema
del trabajo asalariado. El éxito de Marx está en su capacidad para aportar al conocimiento en
torno a las determinaciones del capital cuyo desarrollo tiene a la acción política de la clase
obrera como medio de realizarse, por lo que debemos buscar una respuesta más consistente
al problema que la pura marxología, avanzando en descubrir las potencias revolucionarias de
la clase obrera que sólo puede sacar fuerza de su propia relación social.
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Izquierdas, 29:251-274, septiembre 2016
1. La reproducción de la fuerza de trabajo y el capital
A nuestro parecer, ninguno de los trabajos anteriores logra ser satisfactorio, bien porque
separan la unidad de la relación social general, abstrayendo a la clase obrera de su propia
condición material, o bien porque a pesar de concebir la unidad, reducen el problema a una
discusión marxológica sin abordar el problema de la subjetividad revolucionaria de la clase
obrera. Es por eso que se vuelve necesaria una tercera posición.
Todas las formas sociales necesitan reproducirse, es decir, necesitan que el proceso de
producción sea continuo (Marx 2002, 695). Esto implica, por lo tanto, que del total social
producido una parte debe ser continuamente reconvertida en medios de producción y otra en
fuerza de trabajo. Sin embargo, esto no pasa de ser una afirmación general, común a todo
modo de producción (Marx 1971, 8).
En el caso del modo de producción capitalista la producción tiene como finalidad la
producción de valor y más específicamente de plusvalor, como forma específica de ampliar
la capacidad de crear esta capacidad para organizar el trabajo social como atributo del
producto material del trabajo anterior (Iñigo Carrera 2013, 12). Este proceso de valorización
se reproduce, en su forma más simple, reproduciendo al obrero, por un lado y al capitalista
por el otro, uno como poseedor de fuerza de trabajo, al otro como dueño de los medios de
producción. De esta manera, al repartir el producto social necesario para volver echar a
andar la producción, el capital dinero transmuta bajo la forma de capital constante y
variable. El primero destinado a convertirse en medios de producción, el segundo, como el
medio de producción más importante de todos: fuerza de trabajo que, una vez es puesta en
movimiento, deviene trabajo vivo.
Al subsumir la circulación a la producción de plusvalor, el fondo de subsistencia
destinado a la reproducción de la fuerza de trabajo se le presenta al obrero como medios de
pago equivalentes a una determinada cantidad de valor intercambiable por medios de vida,
los que se le aparecen como un enorme cúmulo de mercancías en la circulación.
Si ponemos nuestra atención a este último proceso, veremos al vendedor de la fuerza de
trabajo que se presenta ante el comprador con la única mercancía que posee para participar
de su relación social general. Al tratarse de una relación social indirecta mediada por
mercancías, la circulación vela la emergencia del plusvalor al aparecer el valor de su fuerza
de trabajo como salario o “valor del trabajo”, quedando oculto a la vista del obrero el origen
mismo de este monto, así como la parte que se embolsa el capitalista, dando la impresión de
que, por un lado, el salario sale del bolsillo del capitalista y la ganancia del intercambio de
mercancías vendidas al final del proceso. Es decir, el monto en dinero que desembolsa el
capitalista y que intercambia por fuerza de trabajo aparece como una mera magnitud que
pertenece al capitalista y que equivale al “valor del trabajo” proporcionado por el obrero.
Pero, dejando de lado la perspectiva individual, el enfrentamiento directo entre vendedor y
comprador de mercancías, el análisis de la reproducción simple hecho por Marx, deja al
descubierto que, detrás del intercambio de equivalentes, donde el capitalista sólo devuelve,
bajo la forma de salario, el trabajo producido por el obrero, se oculta la apropiación del
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plustrabajo creado en el proceso de producción que pone a funcionar con la fuerza de trabajo
bajo su dominio, como un atributo del capital (Marx 2002, 695-713).
Este proceso de compra y venta -que se repite toda vez que el obrero se enfrenta al
capitalista como poseedor de dinero- ocurre bajo ciclos más cortos que la circulación en
general. A esta circulación Marx la denomina “pequeña circulación” (Marx 2005, 195 y
sgts.). Dada la especificidad del modo de producción capitalista, la mercancía fuerza de
trabajo, su circulación, se diferencia de la circulación en general, la que comprende todo el
periodo en que el capital emerge y regresa al proceso de producción. A diferencia de esta
última, la “pequeña circulación” “es continua y ocurre constantemente de manera simultánea
con el proceso de producción. Es la parte del capital que se paga como salario, que se
intercambia por la capacidad de trabajo” (Marx 2005, 195). De esta manera, la pequeña
circulación es la que pone al capital en cuanto tal, al someter a la fuerza de trabajo a la
valorización.
Ahora bien, la repetición constante de este proceso, supone la reproducción del obrero
como algo a parte de las condiciones objetivas del trabajo, lo que conlleva a la separación
del producto del trabajo y el trabajo mismo, condición fundamental del proceso capitalista
de producción. Dicho de otra forma, en tanto proceso que se reproduce, “el capital produce
en su reproducción sus propias condiciones” (Marx 2005, 198) lo que implica no sólo el
destinar una parte del producto para la creación de más medios de producción (capital fijo),
sino la fuerza de trabajo que, puesta junto a los medios de trabajo, los revive dando paso a la
producción de plusvalor.
Desde esta perspectiva, al ser la relación social general la que reproduce sus condiciones
de producción, el consumo del obrero adquiere un sentido muy diferente al que se nos
presentaba desde la perspectiva del obrero individual. Ahora, considerando el proceso de
reproducción en su conjunto, bajo sus determinaciones más simples, “la clase capitalista
entrega constantemente a la clase obrera, bajo la forma dineraria, asignados sobre una parte
del producto creado por esta última clase y apropiado por la primera” (Marx 2002, 697).
Pero así también, “el obrero devuelve a la clase capitalista esos asignados y obtiene de ésta,
así, la parte que le corresponde de su propio producto. La forma mercantil del producto y la
forma dineraria de la mercancía disfrazan la transacción.” (Ibíd.).
Si consideramos sólo el proceso de producción, el consumo del obrero aparecía como
un proceso que se realiza por fuera de la circulación, como consumo individual o
improductivo, por “fuera del capital”. Ahora, desde la perspectiva de la reproducción, este
aparece como improductivo solo para él, mientras que, desde la reproducción de la relación
social general, aparece productivo para el capital, no como plusvalor, sino como la
renovación constante de la valorización, como condición de sí mismo (Marx 2002, 198)..
Por lo tanto, si bien el obrero puede consumir improductivamente su salario, el capital
lo consume a él de manera productiva, por lo que el consumo del obrero aparece como un
momento necesario de la propia valorización y no, como pretenden algunos, como
autovalorización del obrero.
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La importancia de subsumir realmente la fuerza de trabajo al capital, radica en que, al
hacerlo, el capital se apropia del proceso humano genérico de trabajo, portado por los
individuos, sometiéndolos a las necesidades de la valorización. La fuerza de trabajo, que
debe ser producida y reproducida, ahora lo hace de una manera específicamente capitalista al
ser el medio por el cual el capital se valoriza. Dicho de otra manera, la capacidad genérica
del ser humano –portada por los individuos- aparece invertida a la conciencia libre de los
individuos como una capacidad social ajena y que los somete.
Su conciencia libre, que media su consumo individual, se muestra desvinculada de la
producción, de la misma forma que su sujeción al conjunto de la clase capitalista se oculta
detrás del contrato de trabajo y la relación con los capitalistas individuales. Sin embargo, la
reproducción y continuación del proceso, sólo reafirma la subordinación real del obrero al
capital, como forma y condición del mismo tal como con los medios de producción. El
obrero, entonces,
se suministra medios de subsistencia, para mantener en funcionamiento su
fuerza de trabajo, de la misma manera que se suministran carbón y agua a la
máquina de vapor, aceite a la rueda, etcétera. Sus medios de consumo son
entonces meros medios de consumo de un medio de producción, y su consumo
individual pasa directamente a ser consumo productivo. Esto, no obstante, se
manifiesta como un abuso accidental del proceso capitalista de producción”
(Marx 2002, 703).
Por lo tanto,
…el consumo individual del obrero sigue siendo también un elemento de la
producción y reproducción del capital, ya se efectúe dentro o fuera del taller, de
la fábrica, etc., dentro o fuera del proceso laboral; exactamente al igual que lo
que ocurre con la limpieza de la máquina, ya se efectúe dicha limpieza durante
el proceso de trabajo o en determinadas pausas del mismo. El hecho de que el
obrero efectúe ese consumo en provecho de sí mismo y no para complacer al
capitalista, nada cambia en la naturaleza del asunto. De la misma suerte, el
consumo de la bestia de carga no deja de ser un elemento necesario del proceso
de producción porque el animal disfrute de lo que come. La conservación y
reproducción constantes de la clase obrera siguen siendo una condición
constante para la reproducción del capital. El capitalista puede abandonar
confiadamente el desempeño de esa tarea a los instintos de conservación y
reproducción de los obreros (Marx 2002, 703-704).
Ahora bien, no solo el consumo del obrero aparece como una condición necesaria y
puesta por el capital mismo, sino que la misma conciencia libre del obrero se descubre como
una forma concreta de su conciencia enajenada, la forma específica que media su
reproducción, y con ello, la reproducción del capital (Marx 2002, 705).
Despojado de los medios para producir su vida, cada vez que abandona el proceso de
producción tal como entra en él: solo poseyendo su fuerza de trabajo, necesitando
reproducirla para volver a subordinarse a otro. Desde esta perspectiva, su consumo
individual es improductivo, no genera plusvalor alguno. Pero desde la perspectiva del capital
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en su conjunto tal reproducción es productiva para el capital en su conjunto. Por lo tanto,
aquello que desde la perspectiva individual parece bajo determinada forma, se invierte al
considerar al capital como una relación social general. Tal como señala Marx,
el consumo individual del obrero es improductivo para él mismo, puesto que
únicamente reproduce al individuo lleno de necesidades, es productivo para el
capitalista y el estado, puesto que es producción de la fuerza que produce la
riqueza ajena” (Marx 2002, 706).
Por lo tanto
Desde el punto de vista social, la clase obrera, también cuando está fuera del
proceso laboral directo es un accesorio del capital, a igual título que el
instrumento inanimado de trabajo. Incluso su consumo individual no es, dentro
de ciertos límites, más que un factor del proceso de reproducción del capital.
Pero el proceso vela para que esos instrumentos de producción autoconscientes
no abandonen su puesto, y para ello aleja constantemente del polo que ocupan,
hacia el polo opuesto ocupado por el capital, el producto de aquéllos. El
consumo individual, de una parte, vela por su propia conservación y
reproducción, y de otra parte, mediante la destrucción de los medios de
subsistencia, cuida de que los obreros reaparezcan constantemente en el
mercado de trabajo. El esclavo romano estaba sujeto por cadenas a su
propietario; el asalariado lo está por hilos invisibles. El cambio constante de
patrón individual y la fictio juris [ficción jurídica] del contrato, mantienen en
pie la apariencia de que el asalariado es independiente” (Ibíd.).
Esta ficción jurídica –que tiene al contrato como su forma más simple (Marx 2002,
103)-, una vez atravesada su apariencia permite ver que “el obrero pertenece al capital aun
antes de venderse al capitalista. Su servidumbre económica está a la vez mediada y
encubierta por la renovación periódica de la venta de sí mismo, por el cambio de su patrón
individual y la oscilación que experimenta en el mercado el precio del trabajo” (Marx 2002,
711-712).
De esta manera, el proceso de producción capitalista ha quedado determinado no sólo
como proceso de producción plusvalor, sino que
El proceso capitalista de producción, considerado en su interdependencia o
como proceso de reproducción, pues, no sólo produce mercancías, no sólo
produce plusvalor, sino que produce y reproduce la relación capitalista misma:
por un lado el capitalista, por la otra el asalariado” (Marx 2002, 712).
Para la mayoría de los autores vistos, este proceso unitario aparece como algo separado
del capital, como el espacio de reproducción del obrero, realizado de acuerdo a sus propias
necesidades (u otras diferentes a la valorización). La clase obrera no es una forma del capital
sino que es dominada por él, como una relación exterior, quedando limitadas a las
apariencias, prisioneras “de las representaciones en las cuales se mueven los agentes del
modo capitalista de producción” (Marx 1972, 3-4).
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Pretender que el capital expresa una “lógica” o una “necesidad” diferente a la de la clase
obrera es justamente romper este proceso reproductivo, es decir, dejar de concebir al capital
como unidad, como la relación social general dentro de la cual los seres humanos producen
y reproducen su vida, transformando así a la clase obrera en una abstracción, dejándola
impotente, privada de sus potencias históricas.
2. ¿Más allá de Marx y de El Capital?
270
Al parecer, los intentos de Negri, Rubel y Lebowitz tienen la intención de colocar en el
centro de la cuestión la lucha de las clases, rescatar la autonomía de la clase obrera y
desmarcarse de las visiones “unilaterales” o “economicistas”. Tanto Lebowitz como Negri,
teniendo como referencia las observaciones de Rubel y otros ex o “post” marxistas, buscan
los límites del capital en El Capital para seguir el rastro al supuesto gran ausente de su obra
magna: los obreros. Sin embargo, pierden de vista que la clase obrera es tal en la medida que
vive subordinada como un atributo al capital, el cual, como relación social general, no sólo
produce plusvalor, sino los seres humanos necesarios para producirlo. La subsunción real
del proceso de trabajo al capital implica no sólo subordinar la fuerza de trabajo, sino el
proceso de vida genérico humano, el que aparece invertido a los obreros enajenado como
una potencia del capital. Es bajo la forma capitalista en que el metabolismo social general se
realiza. Nos enfrentamos entonces a que el corazón de la clase obrera late al ritmo del
mismo capital.
Al considerar que la relación que establece la clase obrera con el capital es “exterior”, se
ven obligados a buscar un “afuera” de la relación social general donde se desarrollen las
potencias subjetivas que, según estos autores, el capital niega, quedando presos de la
conciencia abstractamente libre. Buscan “salir del capital” por medio del análisis del salario
al momento de determinar la relación entre la parte necesaria y no retribuida de la jornada y
suponen que su contenido es una abstracta lucha de clases. Su misma preocupación por los
asuntos de la clase obrera termina por negarle a esta su propia condición de ser, al separar el
desarrollo de su subjetividad revolucionaria de su propia condición material, negándole su
potencial revolucionario.
Tanto Lebowitz como Negri esperan que la clase obrera alcance una fuerza de tal
dimensión que pueda invertir la situación y ser ella quien domine al capital en el mejor de
los casos, o bien, se limite a ser una pura multitud impotente que quiere hace la revolución
sin tomar el poder, en el peor. En lugar de ser la forma concreta en que la relación social
indirecta mediada por mercancías recobra su unidad bajo la forma política, la lucha de
clases, se reduce a una simple relación exterior, como el choque de dos fuerzas que
encuentran su necesidad de manera contrapuesta, de forma indeterminada. Es decir, como
intereses antagónicos simplemente inmediatos, sin poder explicar lo que expresan como
formas concretas en que se realizan las formas más abstractas que contienen.
El problema de un libro que trate sobre una teoría del salario ha pasado de ser un tema
“filológico” a un problema político sobre los intereses objetivos de la clase obrera, es decir,
sobre las potencias que impulsan sus luchas, la materialidad que contienen. Suponiendo que
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“no se trata de lo que directamente se imagine tal o cual proletario, o incluso el proletariado
entero. Se trata de lo que es y de lo que históricamente se verá obligado a hacer por ese ser”
(Marx y Engels 1967, 102). Aquellas ciencias sociales que pretenden justificar a la clase
obrera como una fuerza “independiente” y contradictoria con el capital la terminan vaciando
de contenido. Para este marxismo “abierto”, será la conciencia la que aparece como un punto
irreductible al capital, como conciencia libre, como mera certeza inmediata de no ser el
capital. Por lo tanto, como ciencia de la apariencia, estas posiciones no pueden ser sino
ideología, expresando un contenido diferente al que dicen portar.
El capital, como forma histórica que asume la necesidad genérica propia de la vida
natural humana, siguiendo su propia valorización, debe reproducir las condiciones que hacen
posible la realización del plusvalor. En tanto forma privada del trabajo social mediado por el
intercambio de mercancías, la forma más simple de dicha relación (la mercancía) que
emerge en el proceso de circulación, se desarrolla como la reproducción constante de la
separación entre obreros y capitalistas, por lo que los primeros no pueden tener más potencia
que la que les entrega su propia relación social. Es decir, el capital, como relación social
enajenada, es condición de su propia superación.
De acuerdo con Marx, la potencialidad de la clase obrera, su carácter revolucionario, no
es una forma contraria al desarrollo del capital mismo, su “antagonista” o su “otro”. Al
contrario, comprender qué es el capital es comprender la forma concreta en que se produce y
reproduce nuestra relación social general, conocer lo que el proceso de producción y
reproducción del capital, como continuo proceso de desarrollo de las fuerzas productivas,
impone fundando las condiciones de nuestra propia acción. Si el capital continúa pujando
por desarrollarse y crecer, buscando abandonar definitivamente su crisálida, deberá
“empujar” a la clase obrera a la lucha revolucionaria. Sin embargo, al adoptar las mismas
potencialidades sociales de forma enajenada, dicha potencia no puede sino aparecerle como
antagónica. Desde tal perspectiva, como sostiene Iñigo Carrera, Marx ha sido el primero en
reproducir en El Capital, esa conciencia enajenada al tanto de su propia enajenación y las
potencias históricas que contiene.8
Sin embargo, esta necesidad histórica se realiza de manera contradictoria. La condición
fundamental de una organización consciente del trabajo es la revolución constante del
proceso de producción, la que se materializa como nuevos medios de producción. Sin
embargo, a diferencia de lo que pasa en su forma manufacturera, bajo el desarrollo del
plusvalor relativo son los medios de producción los que subordinan al obrero como un mero
apéndice. El desarrollo de la capacidad genérica humana del trabajo bajo su forma
capitalista, es decir, como un medio para la valorización, se expresa como una permanente
8
Según Juan Iñigo, “El Capital de Marx es en sí mismo el desarrollo, realizado por primera vez y puesto bajo
una forma que permite su reproducción social, de la conciencia enajenada de la clase obrera que se produce a
sí misma como una conciencia enajenada que conoce su propia enajenación y las potencias históricas que
obtiene de ella. En El capital, esta conciencia se despliega hasta alcanzar sus determinaciones generales que
conciernen a la acción revolucionaria de la clase obrera en la que dichas potencias históricas se realizan
produciendo las condiciones materiales para la organización consciente –por lo tanto, libre– de la vida social”
(Iñigo Carrera 2013, 41-42).
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expansión de esta capacidad genérica, desarrollada como una serie de trabajos concretos
distribuido en un número de capitales individuales por medio de los cuales se realiza la
valorización. Estas formas concretas, ahora formas del capital, se desarrollan como
expresiones fragmentadas de la búsqueda de plusvalor relativo, como competencia entre
capitales. La misma búsqueda de plusvalor impacta en el desarrollo de las fuerzas materiales
que determinan la potencia de la clase obrera, pero también a la clase capitalista, cada una
como personificación del propio desarrollo de esta relación social general. Al ser una
totalidad fragmentada en una serie de unidades que producen privadamente, se ve en la
necesidad de realizarse como relación social directa que establece las clases que personifican
esas potencias, es decir, bajo la forma de lucha de clases y hace de la acción política de la
clase obrera condición de realización de sus potencialidades y consiguiente superación.
La valorización, el impulso que mueve al capital como conjunto, desarrolla las fuerzas
productivas a niveles nunca antes vistos, concentra el capital y coloca a la clase obrera como
el único sujeto capaz de seguir desarrollando tales potencialidades. Ella es, no sólo la
condición de su reproducción y desarrollo, sino que, por ser aquello que es, se vuelve
palanca de su superación. Por lo tanto, lejos de ser una pura fuerza abstracta, la clase obrera
se vuelve un eslabón en el proceso natural humano que se desarrolla por medio de la
apropiación del entorno y, con ello, se desarrolla a sí mismo, teniendo cada vez más su
propia acción consciente como forma necesaria de realización de tales potencialidades.
Como señala Marx casi al final del Tomo I,
No bien ese proceso de transformación [que impone la propiedad capitalista,
que depende del trabajo asalariado, G.R.] ha descompuesto suficientemente,
en profundidad y en extensión, la vieja sociedad; no bien los trabajadores se
han convertido en proletarios y sus condiciones de trabajo en capital; no bien
el modo de producción capitalista puede andar ya sin andaderas, asumen una
nueva forma la socialización ulterior del trabajo y la transformación ulterior
de la tierra y de otros medios de producción en medios de producción
socialmente explotados, y por ende en medios de producción colectivos, y
asume también una nueva forma, por consiguiente, la expropiación ulterior
de los propietarios privados. El que debe ahora ser expropiado no es ya el
trabajador que labora por su propia cuenta, sino el capitalista que explota a
muchos trabajadores.” (Marx 2002, 952-953).
La necesidad de reducir el trabajo necesario al mínimo, ampliando el tiempo de
plustrabajo y buscando la máxima valorización, transmuta por medio de su reproducción
ampliada hacia una forma diferente de organizar la producción social total. La valorización
sienta las bases de su propia desaparición. El capital, como forma de organizar el trabajo
social total abre paso a una nueva forma concreta, una nueva relación social general. Esta no
puede provenir del exterior, producto de una impredecible lucha de clases cuyos avances y
retrocesos parecen condicionar los ciclos de acumulación. El proceso social de producción
que se realiza como su opuesto, como conjunto de trabajos privados, transmuta, siguiendo su
necesidad, en la organización directa de la producción social total bajo la forma de la lucha
de clases.
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De todas maneras, el capital como su propia negación, continuamente busca apropiarse
de la subjetividad productiva del obrero, objetivándola en máquinas que desplazan a más
obreros; así también, utiliza todo el tiempo liberado producto de la reducción del trabajo
necesario como tiempo para crear plusvalor, intensifica la explotación del trabajo e
incrementa la población sobrante desplazada por la misma acumulación. Pero eso no
contradice que la acción política de la clase obrera sea la forma en que se realizan sus
potencialidades, es la forma concreta del mismo desarrollo de aquellas.
Lamentablemente la muerte impide a Marx seguir avanzando en el análisis sobre las
formas desarrolladas del valor: Estado y mercado mundial, el primero como forma concreta
que toma la inserción en el segundo, y sobre las formas que adopta la lucha de clases a partir
del desarrollo de estas determinaciones. Sin embargo, reproducir tales formas hasta dar con
la acción de la clase obrera como la manifestación concreta de realización de dichas
potencialidades, no es problema de Marx, sino nuestro. Centrar la discusión en qué habría o
no dicho Marx al respecto de tal o cual forma desarrollada, no tiene sentido, salvo que sea
como un acto de reconocimiento social para trazarnos un punto de partida original. Sin
embargo, su necesidad sólo se explica abordando estas mismas formas concretas, avanzando
sobre las apariencias que enfrentamos, única forma, por lo demás, de verificar la vigencia de
todo el recorrido hecho hasta acá. En otras palabras, se trata de avanzar en el conocimiento
dialéctico de nuestra propia acción política por medio de la apropiación virtual de las
contradicciones de la realidad actual (Iñigo Carrera 2013, 274), dejando de buscar la
necesidad de la lucha de la clase obrera “por fuera” del capital.
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Smaller limbic structures are associated with greater immunosuppression in over 1000 HIV-infected adults across five continents: Findings from the ENIGMA-HIV Working Group. 2021. ⟨hal-03063822⟩
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bioRxiv preprint doi: https://doi.org/10.1101/724583; this version posted August 5, 2019. The copyright holder for this preprint (which was not
certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under
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Essays on inequality, social preferences and consumer behavior. Economics and Finance. Institut d'études politiques de paris - Sciences Po, 2017. English. ⟨NNT : 2017IEPP0004⟩. ⟨tel-03455045⟩
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The linear expenditure system makes the assumption of separability across commodities through its additive form, which implies independent wants across commodities. This feature is more reasonable when goods are aggregated in broad categories, as substitutes are very imperfect, so we would expect the model to perform better on aggregate groups of commodities (Pollak and Wales, 1969; Pollak, 1971; Deaton, 1975). We gather all items in nineteen categories as indicated by the National Sample Surveys: cereals, footwear, spices, etc. It is also unlikely that this assumption affects our estimates of social subsistence once we control for local own price variations. Nonetheless, we perform the NLP estimation to make sure that cross-price effects do not invalidate our results. Second, the linear expenditure system exhibits linear Engel curves (constant marginal budget shares): the individual purchases necessary quantities of the goods and then divides his supernumerary income among the goods in fixed proportions. Linearity is in fact a good approximation of the Engel curves for below-poverty line households as shown by the non-parametric Engel curves drawn in section 3.5.6.
3.4.2 Empirical Results 3.4.2.1 Simple Demand System: γi
Using the linear expenditure system described in section 3.2.3, we structurally estimate monthly subsistence levels of consumption γi for nineteen categories of expenditure. For all food items, we convert quantities into thousands of calories to have the same quantity unit and ease the conversion into a caloric cost. The sample is restricted to below poverty line (BPL) households in the analytical results that follow. The estimation method used is the iterative generalized nonlinear least square estimator, which takes into account the fact that the demand functions form a complete system (detailed in section 3.4.1). For each expenditure category i, we compute price indexes as described in section 3.3.3, following the method of Deaton and Tarozzi (2000). We estimate the expenditure functions as in Equation (3.11). This specification gives us the following demand system to estimate on n−1 goods for household h in village v and cross-section 106 y:
P x1h,v = β1 mh + γ1 p1,v − β1 i γi pi,v... x n−1h,v = βn−1 mh + γn−1 pn−1,v − βn−1 (3.13) P i γi pi,v
With γi = γi,83 + bi,88 I88 + bi,93 I93 + bi,99 I99 + bi,04 I04 a vector constituted of an intercept and four round fixed effects (1983 is omitted). We add these good-specific NSS round fixed effects in the subsistence level to capture any round-specific variation. The identification of the parameters come from the household-level income variation mh and the village-good-level price variation pi,v. As the sum of expenditure is equal to total expenditure mh, we estimate n−1 equations which give us n parameters γi and n−1 parameters βi (we drop fuel and light expenditure in all estimated systems – the estimation method is not sensitive to P the dropped category). We then compute the parameter βn using the constraint i βi = 1, and Total subsistence pa n ac co tob fue l alc oh ol fru it dry _fr uit me at da iry sp ice pro ce sse d dri nk s clo thi ng _ m clo thi ng _n o foo twe ar fat lse pu ce rea l ve ge tab le su ga r % of mean per capita total expenditure 0 5 10 15 20 25 the parameters γi,y = γi,83 + bi,y for all rounds beside 1983. 95% confidence interval Figure 3.1
:
Total subsistence expenditure
by
broad
categories
(% of
mean
total per capita expenditures)
We take into account the endogeneity of prices by using median village price indexes for all categories i instead of household unit values, following Atkin (2013).8 Villages or urban units are small units in which all households are likely to buy goods at a single market, or consume home-produced goods priced at market level in the NSS data. The measure of total expenditure used to estimate the demand functions is the per capita expenditure on the twenty categories. 8 Atkin (2013) notes that “median village prices are robust to outliers and are not contaminated by quality effects that typically overstate the price response.” 107 The estimation results produce all βi bounded between 0 and 1, and almost all γi positive, as can be seen in Table 3.9 of appendix 3.8.3. The negative γi s correspond to categories with low or zero expenditures and allow the system to be defined at zero. Each estimated subsistence quantity γi is then multiplied by average price and divided by the mean total per capita expenditure. These estimates give an intuitive interpretation of subsistence as a share of total expenditure. Results are presented in Figure 3.1 for Below Poverty Line (BPL) households. In Figure 3.1, we can see that cereal is the first group of expenditure in terms of subsistence, representing more than 20% of the mean monthly total expenditure of BPL households. Then come other caloric items such as fat and pulse, meat, and non-caloric items such as clothing. 0.01 Density.02.03.04 Fuel and intoxicants have very low subsistence expenditure levels. 0 50 100 150
Subsistence expenditure as % of total per capita expenditure, BPL households
Figure 3.2: Subsistence Expenditure as Share of Total Expenditure
sub
sistence
Figure 3.16 in appendix 3.8.3 shows the same results excluding cereal, where we see that other subsistence levels do not exceed 2% of mean monthly per capita expenditure.
Figure
3.17
in appendix 3.8.3 disaggregates the results across rounds by broad categories. Subsistence levels are consistent across rounds, though they show an interesting pattern for cheap calories (cereals, pulses, fat and sugar) whose subsistence level decreased over time. This result shows an interesting trend coherent with the hypothesis of Deaton and Drèze (2009) on the Indian calorie consumption puzzle: a better epidemiological environment and a decreased physical requirement in occupations may explain part of this trend. To explore how subsistence expenditure weight in the per capita total expenditure of the poor, we draw subsistence expenditure as a percentage of total budget for our sample of all NSS rounds (Figure 3.2). The majority of our sample of BPL households is well above the 108 subsistence expenditure level, with a peak at around 30% of the per capita budget. Though comprising a significant share of the budget of the poor, total subsistence expenditure can be afforded by most households in our sample. As a further test on our measure of subsistence expenditure, we sum subsistence quantities for all food categories (γi by rounds in Table 3.9 of appendix 3.8.3), multiply this sum by 1000 to obtain number of calories (recall that the quantity is expressed in thousands of calories) and divide by 30 to obtain the daily per capita subsistence level of calories. We obtain a subsistence level of between 500 (NSS 61st round) and 900 (NSS 38th round) per capita calories, which is usually considered as a lower bound for metabolic survival.9 All these findings are reassuring on the interpretation of these measures as “subsistence” expenditure. 3.4.2.2 Demand System with Relative Deprivation: γi = τi + νi ρ
We disaggregate the subsistence level into an intercept (basic subsistence) and the measure of aggregate relative deprivation ρ which is the Gini index, as derived in section 3.2.1. The Gini index of per capita expenditure in each NSS region provides a local variation in the level of social subsistence. We also add a dummy for urban households and the log of household size, allowing to take into account demographic effects commonly found in demand estimation. The expression of the subsistence parameter γi of Equation (3.13) in this specification is:
γi = τi,0 + νi Ginir + τi,1 Uh + τi,2 ln(size)h + γi,83 + X bi,y Iy (3.14) y6=83
Social subsistence is good-specific, and is composed of the Veblen coefficient νi and the aggregate measure of relative deprivation Ginir. This decomposition allows to test Implications 1, 2 and 3 presented in section 3.2. U is a dummy capturing whether the household lives in an urban area, and ln(size) is the log of the household size. The effect of each of these demographic variables is assumed to depend on each good i, and is captured respectively by parameters τi,1 and τi,2. The remaining parameter τi,0 capture the residual component of subsistence quantities. The specification also contains good-specific year dummies to capture any trend specific to each survey. Figure 3.3 presents the social subsistence levels obtained by Specification (3.14) for all goods as a percentage of total monthly per capita expenditure. To obtain subsistence expenditure, we multiply their Veblen coefficient nui by the mean regional Gini coefficient Ginir and price index. We then divide by the mean monthly total per capita expenditure to have an intuitive estimate of its magnitude. Figure 3.18 in appendix 3.8.3 shows the same results for a specification without the demographic variables. The sign of νi gives us information on socially inferior or socially superior goods (Implication 9 The National Institutes of Health’s Medline Plus considers that a diet of 500 to 800 calories a day is close to starvation. Several clinical experiments involved diets at 500 to 800 calories a day (Bortz, 1969; Ball et al., 1970; Sandhofer et al., 1973; Willms et al., 1978). 109 4 % of mean per capita total expenditure -6 -4 -2 0 2 fue l ar oh ol pro ce sse d clo thi ng _m dri nk s sp ice twe alc foo co _n o ac ng clo thi pa n tob fat su ga r pu lse da iry fru it ge tab le ve ce rea l me at dry _fr uit -8 Social subsistence (cal.) Social subsistence (non-cal.) 95% confidence interval
Figure 3.3: Social subsistence expenditure (% of total expenditure), BPL households 1).
Here, consistently with our hypothesis, cereal is clearly an inferior good, i.e. whose subsistence level decreases with relative deprivation. More interestingly, meat is considered socially inferior as well. This result is a good test of our theoretical definition of inferior and superior goods: in India, meat is a cheap source of proteins as it is considered to make one impure – specifically beef and pork meat. It is therefore reserved to lower sections of the society such as Scheduled Castes, or other religions such as Muslims and Christians. The fact of not consuming meat is a sign of wealth and status, and one of the first practices to be given up in the process of mimicking higher status groups (sanskritization, as defined by Srinivas (1956)). If, in other societies, we would expect meat to be a superior good, it is revealing that the data show the contrary in the case of India. We expect the social standard of meat consumption to decrease with inequality. The socially superior goods are food items associated with wealth and abundance (sugar, fat, drinks, processed food), vegetarianism norm (pulse, dairy products) and non-caloric visible items (clothing, footwear, fuel and light). Apart from alcohol, intoxicants do not respond much to relative deprivation. This result is another interesting outcome of our detection of superior goods, as the consumption of intoxicants has often been underlined as a sign of lack of self-control (temptation goods), and a threat to long-term investments such as nutrition or education. Intoxicants, aside from their addiction and temptation components, are also social goods. Here, additionally, the force of substitution between inferior and superior goods does not rely on them. These results show that, aside from temptation, the social constraint of the poor may also be a plausible explanation for their spending choices. Figure 3.3 gives an intuition of these social subsistence levels as a percentage of monthly 110 n co pa ac tob fue l alc oh ol pro c es se d dri nk s clo thi ng _m clo thi ng _n o foo twe ar iry at ice sp da me fru it fat lse pu ce rea l ve ge tab le su ga r % of mean per capita group expenditure -50 0 50 100 Social subsistence (cal.)
95%
confidence interval Social subsistence (non
-
cal
.) Figure 3.4: Social subsistence expenditure (% of good expenditure), BPL households total budget. These goods, however, have different budget shares – cereals are much more largely consumed than meat, for example.
To give an idea of how important social subsistence is within the good budget, we draw Figure 3.4 which
shows
social subsistence level of good i as a percentage of monthly per capita expenditure on good i. The category of dry fruits is excluded as it is an outlier (around 150% the mean category expenditure), likely due to the very small budget share spent on dry fruits by BPL households in our data. The social subsistence level for cereal now appears to be a small fraction of cereal expenditure (15%). Cereals are the major source of calories for BPL households, so it is not surprising that these households cannot substantially decrease their consumption of cereals. We also see in Figure 3.4 that non-caloric superior goods (darker bars) have on average a social subsistence level comprising a higher share of the category budget than caloric superior goods. This is especially true for spices, drinks and alcohol. Social subsistence for meat, as expected by the social norm of vegetarianism, comprises a bigger share of the budget allocated to this category (around 28%) than cereals. Implications 2 and 3 provide a definition of aspirational goods, i.e. socially superior goods whose demand increases (and income elasticity decreases) with relative deprivation. This definition does not depend solely on the social valuation of the good νi, but also on the social valuation of other goods and the relative budget share (section 3.2.3). We identify the goods qualified as aspirational in our sample by computing income elasticities in regions with different Gini coefficients (Gini of 0.2 in low inequality regions, and 0.4 in high inequality regions – the median Gini is 0.3), but using the same parameters, income and price levels. pan meat dry_fruit cereal drinks tobacco clothing_m spice footwear fat sugar clothing_no alcohol pulse vegetable fuel fruit processed dairy 0.5 1 1.5 2
Income elaticity Low regional inequality High regional inequality Figure 3.5: Estimated income elasticities in low vs. high Gini regions, all rounds Figure 3.5 shows the income elasticities for each good in low (light) and high (dark) inequality regions. We find results close to the ones given by the Gini coefficients nui : cereal and meat are non aspirational goods, as well as most intoxicants (pan, tobacco) and slightly vegetable. On the contrary, goods identified as highly socially superior (sugar, spice, drinks, processed food, dairy, footwear, fuel and light) are clearly aspirational as well. Alcohol seems to be aspirational too.
When relative deprivation increases, socially superior goods tend to become more necessary to the poor. An additional hypothesis, linked to the work of Heffetz (2011) on income elasticities, is that goods which signal status for the wealthier sections of society are goods which are aspirational for the poor. Heffetz (2011) defines signaling goods as luxury goods, i.e. whose income elasticity is higher than one. Indeed, richer individuals allocate a higher share of their budget on such goods to signal their position in society. In a high inequality region, the top income households are wealthier and thus spend more on such goods. In the case where the social standard of consumption is determined by relative deprivation, we would expect that goods classified as luxuries are aspirational, and thus tend to become more necessary in high inequality regions. We see that this is the case in our data: in high inequality regions, luxuries are more necessary to the poor than in low inequality regions (Figure 3.5). Some aspirational goods even reverse, from an income elasticity higher than one in low inequality regions to an income elasticity lower in high inequality regions (spice, footwear, sugar, pulse). Non-aspirational goods, on the contrary, have an income elasticity which is almost always below 1. These results provide an interesting interpretation on social valuation of goods, and hopefully would lead to additional work on the social determinant of consumption over the entire income scale.
3.4.3 Caloric Cost of Relative Deprivation
The Indian poverty line is computed such that the households living below cannot afford a basket of goods which provides adequate nutrition.As shown by Table 3.3, more than 90% of the population living below poverty line is under malnutrition. This fraction does not seem to reduce with time, consistently with the caloric consumption puzzle underlined by Deaton and Drèze (2009) using the same data. BPL households in India would all benefit from a higher calorie consumption. The constraint of social inclusion weights even more heavily on these households when it does not require the same types of goods than the ones which could better their nutrition state.
Fraction under malnutrition Mean daily per capita calories 1983 0.90 1727.31 1988-89 0.90 1742.97 1993-94 0.93 1700.72 1999-00 0.95 1661.93 2004-05 0.97 1623.22 Total 0.93 1685.47
Malnutrition is measured as total daily calories per capita below 2100 (urban) or 2400 (rural). Total calories are computed by multiplying each reported quantity by a nutrient equivalent given by the NSS databases. Table 3.3: Malnutrition among below poverty line households (NSS Data)
To have an order of magnitude of the cost of relative deprivation, we quantify the average loss in consumed calories driven by inequality. From Equation (3.10), we compute the difference in quantity driven by relative deprivation for each good. We think ot this difference as the gap between an individual who does not suffer from relative deprivation or, alternatively, lives in a society where the capability to appear in public without shame is not translated in the commodity space. Intuitively, it is proportional to the gap between two Engel curves with and without relative deprivation, as depicted in Figure 3.9. We can write this gap as the difference between the demand functions with and without relative deprivation. For each good j, it is given by the expression:
P P ∆i = (τi + νi ρ)pi + βi (m − j (τj + νj ρ)pj ) − τi pi + βi (m − j τj pj ) P = νi ρpi − βi j νj ρpj
Section 3.4.2 provides the parameters βi and νj for all goods in the relative deprivation specification. We use the parameters estimated in the model with demographic controls, but results are extremely similar without them. We use the variables computed at regional level: ρ is the per capita expenditure gini by region used in the estimation, and the price index pi is taken at region level. We compute ∆i for each good i using these parameters and variables. In all estimations, quantities have been converted in thousands of calories using the nutrient equivalent for each food item available within the National Sample Surveys. This nutrient equivalent provides the caloric content of all specific items, including drinks, spices, pan or alcohol. The total caloric cost κcalorie is the sum of these calorie differences ∆i for all good i: 113 κcalorie = X ∆i (3.15) i The measure of calorie consumption affected by relative deprivation is not a cost by construction, as it takes into account the social valuation of all caloric items. If caloric items were mostly socially valued, our measure would provide a caloric benefit to relative deprivation. Even though this result would be counter-intuitive, it underlines the flexibility of our framework to account for all aspects of social valuation, letting the empirical analysis determine how each 100 Daily per Capita Calorie Loss 200 300 400 good is affected by relative deprivation.
.2.25.3.35 Gini Coefficient of Monthly Per Capita Expenditure, Region Level.4 Figure 3.6: Calories Forgone in Function of Regional Inequality, BPL households
As our estimation is based on monthly per capita consumption, we divide κcalorie by 30 in order to obtain the average daily per capita caloric loss estimated by our model of relative deprivation. Figure 3.6 shows the calories forgone by below poverty line households in each round when introducing inequality in consumer demand. The caloric loss goes from about 100 to 200 daily calories per capita for a regional Gini of 0.2 to 200 to 350 for a regional Gini of 0.4, which is a substantial amount for malnourished people. Additionally, the caloric cost has increased over time, consistent with the Indian caloric consumption puzzle underlined by Deaton and Drèze (2009). We can also obtain an estimate of the fraction of households whose per capita daily caloric consumption would be above the malnutrition thresholds in the absence of relative deprivation. We add the estimated caloric loss to total calorie consumption for each region within each round, and find that malnutrition would be reduced by around 10 percentage points in the absence of relative deprivation (Table 3.4). The mean daily per capita calories consumed would also be 114 Fraction under malnutrition
Fraction under malnutrition w/o rel. depriv. Mean daily per capita calories Mean daily per capita calories w/o rel. depriv. 1983 0.90 0.82 1726.92 1905.32 1988-89 0.90 0.79 1742.58 1968.28 1993-94 0.93 0.84 1700.73 1915.43 1999-00 0.95 0.89 1661.94 1859.83 2004-05 0.97 0.88 1623.29 1897.62 Total 0.93 0.84 1685.28 1907.93
Table 3.4: Estimated malnutrition among below poverty line households without relative deprivation (NSS Data) much closer to the malnutrition threshold. The estimated caloric loss is an important indicator that relative deprivation is not neutral to the way consumers allocate their budget
.
We interpret these results as a strong clue that it is more expensive for households to reach adequate nutrition in places where relative deprivation is higher.
3.5 Robustness Checks 3.5.1 Non-Linear Preferences
The non-linear preferences demand system is a generalization of the LES relaxing the assumption of independent wants across commodities. It therefore contains all cross-price terms for each demand equation (see section 3.2.2). We estimate the NLP demand system with expenditure on each good i being defined as Equation (3.20) (appendix 3.8.1.1). We use the same database and methodology as for the LES estimation. Figure 3.19 in appendix 3.8.3 presents the social subsistence levels of the NLP estimation compared to the LES estimation.
It is remarkable that for most goods, the estimates are not significantly different. Also, the sign of the Veblen coefficient, giving us information on the social valuation of the goods, is the same except for fat. If the addition of cross-price terms, allowing for substitution between goods, may affect the basic subsistence level for own good τii, it is unlikely to affect directly the social component of consumption. Indeed, the valuation of each good is not linked to the economic environment. Theoretical works have underlined that inequality could affect relative prices if necessary and luxury goods share the same input of production (Dasgupta and Ray, 1986; Baland and Ray, 1991). In our demand system, the local price variation fully accounts for this effect. We find that social subsistence is mostly not affected by these patterns. 3.5.2 Village versus Regional Gini
When considering relative deprivation, we may wonder what the adequate geographical level of analysis is. Does relative deprivation decrease or increase with the geographical unit we take? Bowles and Park (2005) suggest two characteristics of Veblen externalities: first, they are typically asymmetrical, i.e. they cascade downwards: the poor look up to the rich. This is 115 consistent with the assumption of a relative deprivation model, in which inequality affects consumption aspirations and the social standard of decency. Second, the influence of the reference group may be substantially independent of its size. Even though our measure of relative deprivation captures an aggregate level of inequality, there could be more weight at the top of the distribution. The level at which individuals compare their income and feel relatively deprived may be much larger than their own street or city, due to the trickle down effects (a small group at the top influences by cascade all sections of income). These characteristics suggest that a wider area, such as the NSS regions, could measure more accurately the real sense of social deprivation and its impact on consumption. Another consideration could argue in favor of a stronger effect at the regional rather than town level: upward-looking preferences may have stronger effects on the consumption of aspirational goods when these are the only status symbols that people observe from the rich. Typically, wealthy elites of one’s region are publicly seen only through local medias or days of festival, and their consumption practices trickle down the entire income range to reach the poorest sections. On the contrary, positive aspirations, as theoretically modeled by Genicot and Ray (2014b), are long-term monetary investments or investments in human capital visible which may be visible only to one’s neighbors. The choice of the wealthier households in terms of education would then not be observed by poor households. The social standard for aspirational goods may therefore be set at a much higher income rank than the one for education. We therefore could expect that a smaller level of aggregation, such as the smallest sample unit containing ten households in our data (a village, or an urban block), may have a lesser effect on social subsistence. We perform the same estimation of the disaggregated subsistence level (Specification (3.14)), but using the Gini coefficient at village level.
Figure 3.20 of appendix 4.4 shows the difference between social subsistence levels as captured by a regional and village variation of the Gini coefficient.
We find that the village Gini indeed lowers the effect of relative deprivation on consumption choices, though the results are maintained in terms of the sign and relative magnitude of the effect. This finding suggests that the area that matters for setting the social standard of consumption is larger than one’s village or town. 3.5.3 Scheduled Caste versus Muslim Social Subsistence
Our specification can also be used to test if it predicts with accuracy what is conspicuous for individuals. India is marked by strong social and religious divisions, and each social group may have its own definition of socially valued goods when relative deprivation increases. For instance, the empirical results of Section 4.4 show that meat is not socially valued in India, which is consistent with the fact that vegetarianism is the norm of the upper castes, which have a higher social status. In fact, several works point out that food practices are at the root of untouchability (Ambedkar, 1948; Rege et al., 2009), and the process of sanskritization involves adopting higher caste practices, especially regarding diet and cooking (Srinivas, 1956). Inversely, this phenomenon is not true for Muslims outside the caste hierarchy, for whom meat 116 is a usual component of their diet as in Western societies. An interesting test of our specification would be to estimate the demand system with relative deprivation on sub-samples of BPL Scheduled Caste Hindus (former Untouchable) and Muslims. We expect that meat is not a socially inferior good for Muslims, and that food items associated with High Caste consumption (dairy products, vegetables, pulses) is more socially superior for (m ) Clo thi ng ice Sp ol oh Alc iry Da at Me lse Pu Ve ge tab le % of mean per capita total expenditure -2 0 2 4 Scheduled Castes. Muslim social subsistence SC social subsistence 95% confidence interval Figure 3.7: Social subsistence for Muslims and Scheduled Caste Hindus, selected categories Figure 3.7 shows the social subsistence levels for Scheduled Castes and Muslims for selected items, confirming this prediction: meat is socially inferior for Scheduled Castes, who in return value vegetables, pulses and dairy products much more when relative deprivation increases
. Inversely,
alcohol
consumption of
Muslims
, which is a taboo in Islam, does not react to relative deprivation, on the contrary to Scheduled Castes. Muslims seem more sensitive to other goods such as clothing. The social valuation of these two groups is however not significantly different for most categories, especially for the negative social valuation of cereals (see Figure 3.21 in Appendix 3.8.3 for all categories). In a newspaper article, Aparna Pallavi (food researcher) writes: “Contemporary urban Dalit food is mostly spicy, heavy on oil-both of which were hallmarks of rich people’s food. The high use of salt, oil and chilli, therefore, is a reaction to the Dalit sense of deprivation” (Livemint, 2016). Our data suggests a similar pattern.
3.5.4 Full Sample Estimation
In all specifications, we consider the aggregate level of relative deprivation (Gini coefficient) as an adequate measure of the feeling of relative deprivation for each BPL household. It allows to 117 have a measure not correlated with household income and exogenous to her consumption choices. Underlying to the relative deprivation concept is the idea that people are upward-looking: their social standards of consumption are determined by wealthier households. We therefore expect that aggregate relative deprivation would have a lesser impact on the full sample including wealthier households than on the sample restricted to Below Poverty Line households. Figure 3.22 in appendix 3.8.3 shows that it is indeed the case in our data: the social subsistence level of most categories is significantly lower for the entire sample than for BPL households. Meat, however, is even more socially inferior – reflecting the norm of vegetarianism among the wealthier sections of society. Fat also switches to socially inferior. Soft drinks are, on the contrary, more socially valued. Overall, these results suggest that relative deprivation weights more heavily on the poorer sections of society which have to strive to reach both adequate nutrition and social inclusion.
3.5.5 Caloric Cost of Relative Deprivation: All Robustness Checks
Our baseline specification evaluated the daily per capita calorie loss due to relative deprivation to around 200 calories. Table 3.5 summarizes the same amount for all robustness specifications. Adding demographic variables (baseline LES) lowers down the calorie loss, it therefore seems necessary to control for the household composition and sector. The estimation using a village Gini and the one performed on the full sample lower the caloric cost of relative deprivation, as underlined in the above sections. All specifications suggest a negative effect of relative deprivation on the nutrition state of the household. Baseline LES w/o demographics NLP IV Expenditure Village Gini Muslims Scheduled Caste Hindus Full Sample Daily Per Capita Calorie Loss -212.37 -422.35 -497.39 -329.60 -73.16 -213.09 -265.08 -127.57
Table 3.5: Mean Calorie Loss due to Relative Deprivation, All Robustness Checks (NSS Data)
3.5.6 Non-parametric Engel Curves
The utility function which yields the linear expenditure system is quasi-homothetic, thus producing linear Engel curves. It is a convenient theoretical assumption allowing aggregation across consumers (Gorman, 1953), though not systematically verified in the data (see Lewbel (2008) for a summary of the literature). In this section, we proceed to draw non-parametric Engel curves in order to check if linearity is a good approximation of the Engel curves for below poverty line households. To compare the Engel curves for various items across waves, we need a factor of conversion in order to have Purchasing Power Parity (PPP) expenditure. The poverty line used by the Indian government gives a monthly per capita expenditure under which a household is considered poor for each sector within a state; we have different poverty lines for rural Punjab and urban Punjab, for example. As the measure is based on prices for a given basket of goods on which the poor spend a majority of their budget, it is a measure of the cost of living for poor people in a sector within a state. We use these poverty lines to derive a PPP conversion factor which is anchored on the 55th round (1999-2000) in the respective sector within each state. We then divide total household expenditure and expenditure by item using this factor of conversion, and obtain equivalent expenditure by sector, state and round. The factor of conversion takes into account different evolutions across sector and state in time, but reassuringly, the variance 0 Monthly Expenditure in clothing (PPP) 0 100 200 300 400 Monthly Expenditure in cereal (PPP) 500 1000 1500 within round is small. 0 1000 2000 3000 Monthly Total Expenditure (PPP), BPL households 2005 1989 1994 2000 4000 0 1983 1000 2000 3000 Monthly Total Expenditure (PPP), BPL households 2005 kernel = epanechnikov, degree = 3, bandwidth = 293.16 1989 1994 2000 4000 1983 kernel = epanechnikov, degree = 3, bandwidth = 345.64 (a) Cereal expenditure (
b
) Clothing expenditure
Figure 3.8: Non-parametric Engel curves across rounds, BPL households Figures 3.8a and 3.8b are kernel-weighted local polynomial regressions of expenditure on monthly total expenditure.10 The Engel curves are drawn using the sample of below poverty line households in the four NSS rounds, while adjusting for the difference in living standard across sector, state and round. They appear fairly linear for below poverty line households, and confirm that the assumption of the linear expenditure system is a good approximation of our data. We
could note the
slight curvature
which
appear
s concave for cereal and convex
for
clothing, consistent with
these categories
being necessit
ies and luxuries respectively. The Engel curves for the other categories used in the demand system present a similar pattern (Figures 3.23 to 3.32 in appendix 3.8.4).
3.5.7 AIDS Functional Form
The model estimation does not accommodate fixed effects which could control for important determinants of consumption. In this section, we present an Ordinary Least Squares (OLS) 10 The lowest and highest percentiles of monthly total expenditure have been truncated from the Engel curves. 119 estimation of the Linear Approximate Almost Ideal Demand System (Deaton and Muellbauer, 1980) introducing additional controls to test if the relative deprivation effect is robust to other specifications. The main source of concern is a systematic difference in supply side parameters correlated with inequality. For instance, the availability and exposure to different goods could vary across states and sectors. To control for these variations, we introduce fixed effects by state, year and sector. Regions may also be characterized by specific tastes due to spatial sorting or agroclimatic conditions, which could be correlated with inequality. We introduce region fixed effects to control for fixed regional components through time (we follow the same regions over all rounds in the NSS). Finally, as the OLS estimation allows to easily accommodate other variables, we introduce other demographic and occupational controls such as household population by age and gender, if the head of household is self-employed, and if he/she works in the agricultural sector. These controls are specified by Subramanian and Deaton (1996) as affecting demand for nutrition. We estimate the following specification:
sihy = τ0i + νi Giniry + β ln mhy + X γj ln Pj,vy + τ1i Xhy + FEs,u,y + FEr + ihy (3.16) j
With Giniry the Gini of region r in the NSS round y, ln mhy log of real income of household h in NSS round y (monthly per capita expenditure divided by Stone price index), ln Pi,vy stone price index for category j, Xhy a vector of demographic and occupation characteristics (log household size, fraction by age and gender, self-employed, agricultural sector), FEs,u,y a fixed effect at the State*sector*year level, FEr a fixed effect at region level (same region across years), and ihy an error term. We perform the estimation on all rounds at a time, hence the introduction of round-specific and region-specific fixed effects.
Table 3.6: Working-Leser Engel Specification with Gini, BPL households, all rounds Regional Gini log per cap expend. Observations Adjusted R2 log prices demographic controls FE state*sector*year FE region food (1) -0.0732∗∗∗ (0.0234) no calories (2) -0.0099 (0.00934) clothing (3) 0.0714∗∗∗ (0.0165) intox (4) 0.0034 (0.0121) fuel (5) 0.0085 (0.0143) 0.0382∗∗∗ (0.00253) 157693 0.466 Yes Yes Yes Yes -0.0048∗∗∗ (0.00110) 157693 0.433 Yes Yes Yes Yes -0.0254∗∗∗ (0.00236) 157693 0.323 Yes Yes Yes Yes 0.0078∗∗∗ (0.000843) 157693 0.087 Yes Yes Yes Yes -0.0159∗∗∗ (0.00143) 157693 0.366 Yes Yes Yes Yes Standard errors in parentheses ∗ p < 0.10, ∗∗ p < 0.05, ∗∗∗ p < 0.01 120 3.6
shows the results on food, clothing and other non caloric categories. It is striking to see that the regional Gini decreases food expenditure in the same proportion as it increases clothing expenditure, so that the substitution seems to be between these two categories. In fact, a back-of-the-envelop calculation with this estimate of the Gini effect on food shows that, for the median BPL household in a region with a Gini of 0.30 (the median Gini in our data), this estimate corresponds to a caloric cost of about 100 daily per capita calories. This number is smaller, but reassuringly close to the estimates produced by the structural estimation of the linear expenditure system (200 to 250 calories for the same Gini). 3.5.8 Inequality and Wealth Level of the Poor
Another potential issue with our estimate of the caloric cost of relative deprivation stems from the fact that BPL households could be wealthier in regions where inequality is higher. For instance, if inequality is higher - i.e. there are more high incomes - in more developed regions, then the poor may be expected to be comparatively richer too. This correlation could lead to an estimated subsistence level for the poor which has a higher proportion of non-caloric items, if they are wealthier and less malnourished.
Table 3.7: Descriptive Regression: MPCE on regional Gini, BPL households, all rounds 1983 × Regional Gini (1) log per cap expenditure -0.542∗∗∗ (0.0997) 1989 × Regional Gini -1.041∗∗∗ (0.0513) 1994 × Regional Gini -0.853∗∗∗ (0.0570) 2000 × Regional Gini -0.746∗∗∗ (0.0608) 2005 × Regional Gini -0.533∗∗∗ (0.0538) 160086 0.860 Yes Yes Yes Observations Adjusted R2 log prices household size FE year*sector Standard errors in parentheses ∗ p < 0.10, ∗∗ p < 0.05, ∗∗∗ p < 0.01
In order to check if this conjecture is indeed realized in our data, we regress the log of the monthly per capita total expenditure of BPL households on the regional Gini index and the other variables of our estimation (prices, household size and sector). Table 3.7 shows the resulting coefficients of this descriptive region: the correlation between the Gini index and the total expenditure of the poor is negative for all rounds. As we could expect, regions where inequality is higher capture a lower wealth level for the poor, and not some other determinants 121 such as a higher level of development. This correlation rules out the development explanation of the bias towards non-caloric goods that the poor have in high inequality regions. 3.6 Short and Long-term Consequences 3.6.1 Measurement of Deprivation
These findings bring empirical evidence to our understanding of poverty as the state of deprivation in multiple dimensions. The methodology used could be extended to identify deprivation of different capabilities, following Sen (1983, 1984)’s approach to poverty. Sen (1983) asserts that “absolute deprivation in terms of a person’s capabilities relates to relative deprivation in terms of commodities, incomes and resources”. This definition leads to an understanding of income not as reflecting command over commodities, but over capabilities. Consumption provides a mean to reach several ends ranging from adequate nutrition to social esteem and decency. In fact, in Sen (1983)’s work, as well as in a long tradition dating from Adam Smith (1776) and his example of the linen shirt, the capability to not appear ashamed in public has been considered of central importance for understanding deprivation.11 The capability approach leads us to consider that an individual is poorer than another if, with the same real income, she cannot attain physical basic needs and social decency. By identifying that households below poverty line consume less calories where the social standard is higher, we may say that these households are deprived of more capabilities than equivalent households in less unequal places. Though we cannot have a utilitarian welfare interpretation of this substitution between food and social commodities – as an individual spending more on social commodities may be as satisfied as another spending more on food –, the capability approach allows us to infer that one is worse-off than the other in terms of reaching several capabilities (meeting nutritional requirements, not being ashamed in public). The second and corollary result is that even under necessity, an individual does not fulfil one capability (for example, adequate nutrition) before others (social decency, self-respect), but weights all of them within her budget constraint. In the literature on poverty line, several works have tried to conceal absolute and relative dimensions of poverty. Atkinson and Bourguignon (2001) derive a poverty line in terms of economic resources combining physical basic needs and socially defined minimum consumption standards. They define these dimensions in the capability space as well, these two needs corresponding to functioning satisfactorily in purely physical terms and in social terms. Ravallion and Chen (2011) propose a weakly relative poverty line, recognizing that the poor in terms of physical deprivation also strives for social inclusion: they underline that “the cost of a sociallyacceptable linen shirt will not be zero, and will presumably be no different for a poor person.” 11 Smith (1776) notes that “the Greeks and Romans lived very comfortably though they had no linen, [but] in the present time, through the greater part of Europe, a creditable day-laborer would be ashamed to appear in public without a linen shirt”. 122 Our work suggests a measure of social need derived from the literature on relative deprivation as the sum of income gaps (rather than the mean). It provides an empirical method to determine how social need affects consumption choices of people who are highly budget-constrained, and an estimate of the cost incurred to fulfill both physical and social needs when the level of the latter is rising. The methodology can be applied to other dimensions of deprivation and other databases, both to confirm these results and better inform on the multiple costs of deprivation.
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Glio‐ and neuro‐protection by prosaposin is mediated by orphan G‐protein coupled receptors GPR37L1 and GPR37
|
Beihui Liu
|
English
|
Spoken
| 12,724
| 25,224
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Liu, B., Mosienko, V., Vaccari Cardoso, B., Prokudina, D.,
Huentelman, M., Teschemacher, A., & Kasparov, S. (2018). Glio‐ and
neuro‐protection by prosaposin is mediated by orphan G‐protein
coupled receptors GPR37L1 and GPR37. Glia. Advance online
publication. https://doi.org/10.1002/glia.23480 Publisher's PDF, also known as Version of record
License (if available):
CC BY
Link to published version (if available):
10.1002/glia.23480 Link to publication record on the Bristol Research Portal
PDF-document This is the final published version of the article (version of record). It first appeared online via Wiley at
https://onlinelibrary.wiley.com/doi/full/10.1002/glia.23480. Please refer to any applicable terms of use of the
publisher. Liu, B., Mosienko, V., Vaccari Cardoso, B., Prokudina, D.,
Huentelman, M., Teschemacher, A., & Kasparov, S. (2018). Glio‐ and
neuro‐protection by prosaposin is mediated by orphan G‐protein
coupled receptors GPR37L1 and GPR37. Glia. Advance online
publication. https://doi.org/10.1002/glia.23480 Liu, B., Mosienko, V., Vaccari Cardoso, B., Prokudina, D.,
Huentelman, M., Teschemacher, A., & Kasparov, S. (2018). Glio‐ and
neuro‐protection by prosaposin is mediated by orphan G‐protein
coupled receptors GPR37L1 and GPR37. Glia. Advance online
publication. https://doi.org/10.1002/glia.23480 MAIN POINTS G-protein-coupled
receptors
(GPCRs)
are
particularly
attractive
because they are the most plausible targets for modern small molecule
drugs, but this approach critically depends on identification of their
endogenous agonists. The search for druggable targets in the brain
conventionally focused on neurons, but astrocytes as natural neuro-
protectors represent particularly attractive drug targets. Their neuro-
protective mechanisms are numerous and include uptake of glutamate
to prevent neurotoxicity, regulation of extracellular ions and pH, pro-
vision of antioxidative molecules (e.g., glutathione) and trophic factors,
control of micro-circulation, etc. (Liu, Teschemacher, & Kasparov, 2017). 1. Prosaptide TX14(A), a fragment of Saposin C, acts via GPR37L1/
GPR37 on astrocytes and protects them from oxidative stress. 2. In HEK293 cells, GPR37L1 and GPR37 are dysfunctional. 3. GPR37L1/GPR37 signaling in astrocytes enables neuroprotection. 1. Prosaptide TX14(A), a fragment of Saposin C, acts via GPR37L1/
GPR37 on astrocytes and protects them from oxidative stress. 1. Prosaptide TX14(A), a fragment of Saposin C, acts via GPR37L1/
GPR37 on astrocytes and protects them from oxidative stress. 2. In HEK293 cells, GPR37L1 and GPR37 are dysfunctional. 2. In HEK293 cells, GPR37L1 and GPR37 are dysfunctional. 3. GPR37L1/GPR37 signaling in astrocytes enables neuroprotection. 3. GPR37L1/GPR37 signaling in astrocytes enables neuroprotection. KEYWORDS Their neuro-
protective mechanisms are numerous and include uptake of glutamate
to prevent neurotoxicity, regulation of extracellular ions and pH, pro-
vision of antioxidative molecules (e.g., glutathione) and trophic factors,
control of micro-circulation, etc. (Liu, Teschemacher, & Kasparov, 2017). In 1994, the peptide prosaposin (PSAP) and its fragment Saposin
C (Sap C) were identified as neurotrophic factors using the neuroblas-
toma NS20 line and specific binding of radio-labeled Sap C with a Kd
Sergey Kasparov and Anja G. Teschemacher contributed equally to this study. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium,
provided the original work is properly cited. © 2018 The Authors. GLIA Published by Wiley Periodicals, Inc. Glia 2018;1–13
wileyonlinelibrary com/journal/glia
1 University of Bristol – Bristol Research Portal
General rights This document is made available in accordance with publisher policies. Please cite only the
published version using the reference above. Full terms of use are available:
http://www.bristol.ac.uk/red/research-policy/pure/user-guides/brp-terms/ Received: 16 February 2018
Revised: 30 May 2018
Accepted: 4 June 2018 Received: 16 February 2018
Revised: 30 May 2018
Accepted: 4 June 2018
DOI: 10.1002/glia.23480 Glio- and neuro-protection by prosaposin is mediated by
orphan G-protein coupled receptors GPR37L1 and GPR37
Beihui Liu1 | Valentina Mosienko1 | Barbara Vaccari Cardoso1 | Daria Prokudina3 |
Mathew Huentelman2 | Anja G. Teschemacher1 | Sergey Kasparov1 1Department of Physiology, Pharmacology,
and Neuroscience, University of Bristol,
Bristol, United Kingdom
2Translational Genomics Research Institute
(TGen), Phoenix, Arizona
3Baltic Federal University, Kaliningrad, Russia
Federation 1Department of Physiology, Pharmacology,
and Neuroscience, University of Bristol,
Bristol, United Kingdom Discovery of neuroprotective pathways is one of the major priorities for neuroscience. Astrocytes
are natural neuroprotectors and it is likely that brain resilience can be enhanced by mobilizing their
protective potential. Among G-protein coupled receptors expressed by astrocytes, two highly related
receptors, GPR37L1 and GPR37, are of particular interest. Previous studies suggested that these
receptors are activated by a peptide Saposin C and its neuroactive fragments (prosaptide TX14(A)),
which were demonstrated to be neuroprotective in various animal models by several groups. How-
ever, pairing of Saposin C or prosaptides with GPR37L1/GPR37 has been challenged and presently
GPR37L1/GPR37 have regained their orphan status. Here, we demonstrate that in their natural habi-
tat, astrocytes, these receptors mediate a range of effects of TX14(A), including protection from
oxidative stress. The Saposin C/GPR37L1/GPR37 pathway is also involved in the neuroprotective
effect of astrocytes on neurons subjected to oxidative stress. The action of TX14(A) is at least par-
tially mediated by Gi-proteins and the cAMP-PKA axis. On the other hand, when recombinant
GPR37L1 or GPR37 are expressed in HEK293 cells, they are not functional and do not respond to
TX14(A), which explains unsuccessful attempts to confirm the ligand-receptor pairing. Therefore, this
study identifies GPR37L1/GPR37 as the receptors for TX14(A), and, by extension of Saposin C, and
paves the way for the development of neuroprotective therapeutics acting via these receptors. 3Baltic Federal University, Kaliningrad, Russia
Federation Correspondence
Sergey Kasparov, Department of Physiology,
Pharmacology, and Neuroscience, University
of Bristol, Bristol, 9 BS8 1TD, UK. Email: sergey.kasparov@bristol.ac.uk
and
Anja G. Teschemacher, Department of
Physiology, Pharmacology, and Neuroscience,
University of Bristol, Bristol, 9 BS8 1TD, UK. Email: anja.teschemacher@bristol.ac.uk
Funding information
Biotechnology and Biological Sciences
Research Council, Grant/Award Number: BB/
L019396/1; Medical Research Council, Grant/
Award Number: MR/L020661/1 Correspondence Correspondence
Sergey Kasparov, Department of Physiology,
Pharmacology, and Neuroscience, University
of Bristol, Bristol, 9 BS8 1TD, UK. Email: sergey.kasparov@bristol.ac.uk
and KEYWORDS astrocyte, astroprotection, cAMP, GPR37, GPR37L1, neuroprotection, orphan receptors, PKA,
prosaptide, Saposin C MAIN POINTS
1. Prosaptide TX14(A), a fragment of Saposin C, acts via GPR37L1/
GPR37 on astrocytes and protects them from oxidative stress. 2. In HEK293 cells, GPR37L1 and GPR37 are dysfunctional. 3. GPR37L1/GPR37 signaling in astrocytes enables neuroprotection. 1
|
INTRODUCTION
Any new target for effective neuroprotective therapy must be actively
explored as it may have major medical and societal impacts. Orphan
G-protein-coupled
receptors
(GPCRs)
are
particularly
attractive
because they are the most plausible targets for modern small molecule
drugs, but this approach critically depends on identification of their
endogenous agonists. The search for druggable targets in the brain
conventionally focused on neurons, but astrocytes as natural neuro-
protectors represent particularly attractive drug targets. Their neuro-
protective mechanisms are numerous and include uptake of glutamate
to prevent neurotoxicity, regulation of extracellular ions and pH, pro-
vision of antioxidative molecules (e.g., glutathione) and trophic factors,
control of micro-circulation, etc. (Liu, Teschemacher, & Kasparov, 2017). In 1994, the peptide prosaposin (PSAP) and its fragment Saposin
C (Sap C) were identified as neurotrophic factors using the neuroblas-
toma NS20 line and specific binding of radio-labeled Sap C with a Kd
Sergey Kasparov and Anja G. Teschemacher contributed equally to this study. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium,
provided the original work is properly cited. © 2018 The Authors. GLIA Published by Wiley Periodicals, Inc. Glia. 2018;1–13. wileyonlinelibrary.com/journal/glia
1 MAIN POINTS
1. Prosaptide TX14(A), a fragment of Saposin C, acts via GPR37L1/
GPR37 on astrocytes and protects them from oxidative stress. 2. In HEK293 cells, GPR37L1 and GPR37 are dysfunctional. 3. GPR37L1/GPR37 signaling in astrocytes enables neuroprotection. 1
|
INTRODUCTION
Any new target for effective neuroprotective therapy must be actively
explored as it may have major medical and societal impacts. Orphan
G-protein-coupled
receptors
(GPCRs)
are
particularly
attractive
because they are the most plausible targets for modern small molecule
drugs, but this approach critically depends on identification of their
endogenous agonists. The search for druggable targets in the brain
conventionally focused on neurons, but astrocytes as natural neuro-
protectors represent particularly attractive drug targets. 2.1.1
|
Astrocytes Primary cultures of astrocytes were prepared from the cerebral corti-
ces, cerebellum, and brainstem from Wistar rat pups (P2) following
protocols described previously (1–3). Briefly, the brains of Wistar P2
pups were dissected out, crudely cross-chopped and bathed in a solu-
tion containing HBSS, DNase I (0.04 mg/mL), trypsin from bovine
pancreas (0.25 mg/mL) and BSA (3 mg/mL). The preparation was agi-
tated at 37 C for 15 min. Trypsinization of the brain tissue was termi-
nated by the addition of equal volumes of culture media comprised of
DMEM,
10%
heat-inactivated
FBS,
100 U/mL
penicillin,
and
0.1 mg/mL streptomycin and then centrifuged at 2000 rpm, at room
temperature (RT) for 10 min. The supernatant was aspirated, and the
remaining pellet was resuspended in 15 mL HBSS containing BSA
(3 mg/mL) and DNase I (0.04 mg/mL) and triturated gently. After the
cell debris had settled, the cell suspension was filtered through a
40-μm cell strainer (BD Falcon, BD Biosciences, Franklin Lakes, NJ)
and cells were collected after centrifugation. Cells were seeded in a
T75 flask containing culture media (see above) and maintained at
37 C with 5% CO2. Once the cultures reached confluence and
1 week later, the flasks were mildly shaken overnight to remove
microglia and oligodendrocytes. When astrocytes were seeded for
experiments, media was changed to DMEM supplemented with 5%
FBS instead of 10% FBS. This is to reduce the content of PSAP in the
culture media hence make PSAP depletion easier to achieve. Note
that there was no difference in cell growth in the media containing
either 5% or 10% FBS. However, the pairing of GPR37L1/GPR37 with PSAP and TX14(A)
(Meyer et al., 2013) was later challenged. It was reported that these
receptors are highly constitutively active and couple via Gs proteins,
rather than the Gi pathway as originally reported (Meyer et al., 2013). Moreover, on the background of their high constitutive activity,
TX14(A) was ineffective (Coleman et al., 2016; Giddens et al., 2017;
Ngo et al., 2017). Regulation of this constitutive activity was sug-
gested to occur via cleavage of the extracellular part of GPR37L1
(Coleman et al., 2016; Mattila, Tuusa, & Petaja-Repo, 2016). These
reports reinforced the skepticism based on the failure of TX14(A) to
activate GPR37L1/GPR37 using the DiscoverX orphan receptor-
screening panel (Smith, 2015; Southern et al., 2013). LIU ET AL. of 19 pM was demonstrated (O'Brien, Carson, Seo, Hiraiwa, & Kishi-
moto, 1994). Soon after it was shown that chronic icv infusion of
recombinant PSAP almost completely prevented ischemia-induced
learning deficits and neuronal loss in gerbils (Sano et al., 1994) and the
existence of a GPCR for the neuroprotective part of Sap C was thus
postulated (Hiraiwa, Campana, Martin, & O'Brien, 1997). The experi-
mental usefulness of PSAP and Sap C is limited by their length but,
luckily, the neuroactive part is rather short and can be mimicked by
peptides known as prosaptides, of which the most studied is prosap-
tide TX14(A). The sequence of TX14(A) is highly evolutionarily con-
served (Supporting Information Figure S1). Although neuroprotective
effects of PSAP fragments were demonstrated in several models
in vitro and in vivo (Campana et al., 1998; Gao et al., 2016; Hozumi
et al., 1999; Otero, Conrad, & O'Brien, 1999), the underlying mecha-
nism remained unclear until 2013, when the two closely related
orphan receptors GPR37L1 and GPR37 (Leng, Gu, Simerly, & Spindel,
1999) were proposed to mediate the actions of PSAP and its mimetics
(Meyer, Giddens, Schaefer, & Hall, 2013). 2 2 astrocytes vigorously respond to stimuli which increase cAMP produc-
tion (such as agonists of Gs-coupled receptors or low concentrations
of forskolin), indicating that their resting levels of cAMP are not any-
where near saturation (see, for example, Clark and Perkins, 1971;
Goldman and Chiu, 1984; Tardy et al., 1981). We hypothesized that coupling of GPR37L1/GPR37 in transiently
transfected cell lines does not reveal their true physiological signaling
and reevaluated them in their natural habitat, the astrocytes. Our find-
ings demonstrate that PSAP is, indeed, the natural ligand of
GPR37L1/GPR37 and pave the way for development of neuroprotec-
tive drugs based on this signaling system. 2.1
| Primary cultures of astrocytes and cortical
neurons Experiments were performed in accordance with the UK Animals
(Scientific Procedures) Act, 1986, and were approved by the Univer-
sity of Bristol ethics committee. GPR37L1 is highly expressed by astrocytes, which also express
low levels of GPR37 (Supporting Information Figures S2 and S3; Jolly
et al., 2017; Marazziti et al., 2007; Smith, 2015; Zhang et al., 2014). GPR37 is highly expressed in dopaminergic neurons and early work
focused on the idea of GPR37 being involved in Parkinson's disease
(Cantuti-Castelvetri et al., 2007; Imai et al., 2001; Marazziti et al.,
2007). The importance of GPR37L1 for brain function has been
recently demonstrated in humans. A point mutation in GPR37L1 leads
to a severe neurological phenotype which includes intractable epi-
lepsy, lethal in some of the affected individuals (Giddens et al., 2017). GPR37L1- and especially double GPR37L1/GPR37-knockout mice
were highly susceptible to seizures (Giddens et al., 2017). Moreover,
the deletion of GPR37L1 drastically increased the neuronal loss after
an ischemic stroke (Jolly et al., 2017). These and other findings under-
score the importance of GPR37L1 for brain health. 1
|
INTRODUCTION Any new target for effective neuroprotective therapy must be actively
explored as it may have major medical and societal impacts. Orphan
vision of antioxidative molecules (e.g., glutathione) and trophic factors,
control of micro-circulation, etc. (Liu, Teschemacher, & Kasparov, 2017). In 1994, the peptide prosaposin (PSAP) and its fragment Saposin
C (Sap C) were identified as neurotrophic factors using the neuroblas-
toma NS20 line and specific binding of radio-labeled Sap C with a Kd
Sergey Kasparov and Anja G. Teschemacher contributed equally to this study. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium,
provided the original work is properly cited. © 2018 The Authors. GLIA Published by Wiley Periodicals, Inc. Glia. 2018;1–13. wileyonlinelibrary.com/journal/glia
1 Any new target for effective neuroprotective therapy must be actively
explored as it may have major medical and societal impacts. Orphan In 1994, the peptide prosaposin (PSAP) and its fragment Saposin
C (Sap C) were identified as neurotrophic factors using the neuroblas-
toma NS20 line and specific binding of radio-labeled Sap C with a Kd Glia. 2018;1–13. wileyonlinelibrary.com/journal/glia 1 2.1.1
|
Astrocytes Importantly, all
studies reporting high constitutive activity of GPR37L1 and GPR37
and lack of TX14(A) agonism relied on expression of recombinant
GPR37L1 in either HEK293 or CHO cells (Giddens et al., 2017; Ngo
et al., 2017; Southern et al., 2013) or yeast (Coleman et al., 2016). Neuron/astrocyte co-cultures Neurons were prepared (see above) and plated at 1 × 105 cells per
well on poly-D-lysine-coated glass coverslips in 24-well plates. Astro-
cyte inserts were prepared by plating astrocytes on poly-D-lysine-
coated cell culture inserts with 1 μm diameter pores (Greiner Bio-One,
Monroe, NC) in the same serum-free media as used for neurons. Astrocyte inserts were introduced into neuronal cultures as required. The separation between both cell types allowed secreted molecules
to freely diffuse while preventing direct astrocyte-to-neuron contact. 2.2
| Real-time PCR on primary cultured and acutely
isolated astrocytes To verify that the expression of GPR37L1 and GPR37 in our cultured
astrocytes is not a result of in vitro conditions, we performed acute
vibro-isolation of cortical astrocytes from P12 rats using a method
recently described by Lalo and Pankratov (2017). Approximately
50 single astrocytes were manually collected from the bottom of a
small Petri dish into a sterile test tube. Power SYBR Green Cells-to-Ct
Kit (Ambion Inc., Austin, TX) was used to reverse transcribe directly
from cultured cell lysates, without isolating RNA. The resulting cDNA
samples were then analyzed using QuantiTect SYBR green PCR kit
(Qiagen, Hilden, Germany) on DNA Engine OPTICON 2 continuous
fluorescence detector, following the manufacturer's protocol. β-Actin
was used as a reference house-keeping gene. All primers were
designed to span at least one intron and to produce products of
~100 bp and prevalidated for their efficiency. Products of PCR reac-
tion were resolved on agarose gel to confirm their sizes (Supporting
Information Figure S3). Sequences of the primers are: β-Actin forward:
CTAAGGCCAACCGTGAAAAG; reverse: GGCATACAGGGACAACA-
CAG; GPR37L1 forward: ATGTTTCTTGCCGAGCAGTG; GPRF37L1
reverse: CCACATGGAATCGGTCTATG; GPR37 forward: TCCAT-
GAGTTGACCAAGAAG;
GPR37
reverse:
CTATGCACAGTGCACA-
TAAG; GFAP forward: GAGAGGAAGGTTGAGTCGCT; GFAP reverse:
CACGTGGACCTGCTGCTG. 2.3
| Western blotting For verification of GPR37L1/37 knock-down with adenoviral vectors
(AVV), transduced astrocytes were harvested and placed on ice and
washed with ice-cold phosphate-buffered saline (PBS). The membrane
proteins were then extracted using the Mem-PER eukaryotic mem-
brane protein extraction reagent kit (PIERCE) and purified with SDS-
PAGE sample preparation kit (PIERCE, Appleton, WI). After quantifica-
tion with BCA protein assay kit (PIERCE), 20 μg of membrane protein
per lane were fractionated on a 4–12% Bis–Tris gel (NuPage 4–12%
Bis–Tris Gel, Life Technologies, Carlsbad, CA), and transferred to a
polyvinylidene difluoride (PVDF) membrane (Millipore, Burlington,
MA). After blocking with 5% nonfat dry milk (NFDM) in Tris-buffered
saline with 0.1% Tween-20 (TBST) buffer for 45 min at RT, the PVDF
membrane was cut into two parts at 100 kDa size level. The part of
the membrane containing small sized proteins was incubated with pri-
mary antibody to GPR37L1 (1:1000 dilution) or GPR37 (1:1000 dilu-
tion) in 3% NFDM-TBST at 4 C overnight, and the other part of
membrane was incubated with primary antibody to pan-cadherin
(120 kDa) (1:2000 dilution) as a membrane protein loading control, in
3% NFDM-TBST at 4 C overnight. Following incubation with horse-
radish peroxidase conjugated secondary antibody (DAKO, 1:2000
dilution) for 90 min at RT, the immunoreactivities were detected with
Immun-Star Western C chemiluminescent kit (Bio-Rad Laboratories,
Hercules, CA). For the PSAP depletion assay and the proof of the exis-
tence of PSAP in serum-supplemented culture media, we used the
same protocol as described above except that 5 μL of media or elution
from protein A magnetic beads was applied as the sample volume. A
polyclonal rabbit anti-PSAP antibody was employed. For Western
blotting of PSAP in neuron–astrocyte co-culture media, we changed
to the Amersham ECL Plex western blotting system using a low-
fluorescent PVDF membrane (GE Healthcare, Chicago, IL) and Alexa
Fluor 488 secondary antibody-conjugated goat anti-rabbit. Protein
transfer and membrane blocking was the same as in the above proto-
col. Membranes were incubated with anti-PSAP (1:500 dilution) over-
night at 4 C. Secondary antibody Alexa Fluor 488 was incubated for
1 hr in the dark at room temperature. Before imaging, the membrane
was thoroughly washed. Signal was detected by scanning the mem-
brane on a fluorescent laser scanner (Typhoon, GE Healthcare). 3 LIU ET AL. 3 into pieces <1 mm3 and dissociated in 0.25% Trypsin in dis-
section saline in the presence of 3 mg/mL BSA at 37 C for 15 min. An equal volume of plating media (Neurobasal A with 5% horse serum,
2% B27, 400 nM L-glutamine, 100 U/mL penicillin, and 0.1 mg/mL
streptomycin) was added to terminate the dissociation. Cells were pel-
leted at 2000 rpm for 5 min at room temperature, resuspended in
plating media, and triturated gently. The cell suspension was diluted
appropriately and passed through a 40-μM cell strainer. Approxi-
mately 1 × 105 cells per well were plated on poly-D-lysine-coated
glass cover slips in 24-well plates. Two hours later, the plating media
was replaced with feeding media (Neurobasal A with 2% B27,
800 nM L-glutamine, 20 U/mL penicillin, and 20 μg/mL streptomycin). On day 5, half of the media was replaced with feeding media in which
glutamine was replaced with 4 μM Glutamax. The antimitotic cytosine
β-D-arabinofuranoside (10 μM) was added to control glial contamina-
tion. Neurons were used for experiments 10 days later. 2.1.2
|
Neurons Thus, the nature of the endogenous agonist of GPR37L1 and
GPR37 is currently elusive. Cerebral cortices were dissected out from a litter (8–12) of Wistar rat
embryos on gestation day 18 (E18) and collected in dissection saline
(HBSS, 25.6 mM glucose, 10 mM MgCl2, 1 mM Hepes, 1 mM kynure-
nic acid, 0.005% phenol red, 100 U/mL penicillin, and 0.1 mg/mL
streptomycin). Meninges were removed, and tissues were chopped High constitutive activity of GPR37L1/GPR37 should lead to per-
sistent production of copious amounts of cAMP. However, this has
never been noticed in astrocytes where GPR37L1 is particularly abun-
dant (Supporting Information Figures S2 and S3). To the contrary, 2.6
| cAMP assays Two types of detection were used. 2.4
| Generation of knock-down AVV AVV for the knock-down experiments were based on a modified Pol II
miR RNAi Expression Vector system (Invitrogen) and our previous
work (Liu, Xu, Paton, & Kasparov, 2010). Three AVV were con-
structed,
namely,
AVV–CMV–EmGFP–miR155/GPR37L1,
AVV–
CMV–EmGFP–miR155/GPR37,
and
AVV–CMV–EmGFP–miR155/
negative. The first two were used for knocking down GPR37L1 and
GPR37 in astrocytes, respectively. The third one is a negative control,
harboring a miRNA sequence that can form a hairpin structure that is
processed into mature miRNA but is predicted not to target any
known vertebrate gene. All three vectors were made based on
BLOCK-iT Pol II miR RNAi Expression Vector Kit with EmGFP
(Invitrogen, Carlsbad, CA). This system supports chaining of miRNAs,
thus ensuring co-cistronic expression of multiple miRNAs for knock down of a single target. Three sets of two complementary single-
stranded DNA micro-RNA sequences (targeting different regions of
the same gene) for GPR37L1 and GPR37 were designed by the
BLOCK-iT RNAi Designer (Invitrogen). The complementary single-
stranded oligos were then annealed and cloned into the linearized
pcDNA 6.2-GW/EmGFP–miR vector. The pre-miRNA expression cas-
settes were transferred into an adeno shuttle plasmid pXcX–Sw-linker
(Duale, Kasparov, Paton, & Teschemacher, 2005), resulting in con-
struction of pXcX–CMV–EmGFP–miR155/GPR37L1, pXcX–CMV–
EmGFP–miR155/GPR37 and pXcX–CMV–EmGFP–miR155/negative. AVV were then produced by homologous recombination of shuttle
and the helper plasmid pBHG10 in HEK293 cells. The media was col-
lected for subsequent rounds of AVV proliferation in HEK293 cells
until cytopathic effects were achieved. AVV were purified using CsCl
gradient protocols. Titers were established using an immunoreactivity
spot assay as described previously (Duale et al., 2005). 4 LIU ET AL. LIU ET AL. 4 Klarenbeek, Goedhart, van Batenburg, Groenewald, & Jalink, 2015;
Klarenbeek & Jalink, 2014) specifically in astrocytes. PDM was
exchanged daily. For recording, coverslips were placed into a cham-
ber on a confocal microscope and continuously superfused with
HEPES-buffered solution (HBS; in mM: NaCl 137, KCl 5.4, Na2HPO4
0.34, KH2PO4 0.44, CaCl2 1.67, MgSO4 0.8, NaHCO3 4.2, HEPES
10, Glucose 5.5; pH 7.4, 31.4 C). After taking baseline readings of
astrocytes in the field of view, cells were exposed to 5 min of
NKH477 (0.5 μM), alone or in combination with TX14(A) (100 nM). Fluctuations in cAMP were monitored through CFP/YFP (465–-
500 nm/515–595 nm bands) emission ratios upon 458 nm light
excitation. Images were acquired every 4 s. All FRET ratios were nor-
malized to baseline. 2.8 A wound recovery assay was carried out to analyze the migration of
astrocytes using the IncuCyte system (Essen BioScience Inc., Ann
Arbor, MI). Primary astrocytes were seeded in ImageLock 96-well
plates (4,379, Essen BioScience) at a density of 4 × 104 per well. After 24 hr, standardized and reproducible (700–800 μm wide)
scratch “wounds” were created in all wells using a dedicated device. Cultures were exposed to different testing conditions, for example,
stressors, and were maintained and imaged at hourly intervals up
to 72 hr. 2.7
| PRESTO-Tango β-arrestin-recruitment assay To measure receptor activation, the PRESTO-Tango β-arrestin-
recruitment assay was performed as previously described, with modi-
fications (Kroeze et al., 2015). HTLA cells, a HEK293 cell line stably
expressing a tTA-dependent luciferase reporter and a β-arrestin2-TEV
fusion gene, were used. The cells were maintained in DMEM media
supplemented with 10% FBS, 100 U/mL penicillin and 100 μg/mL
streptomycin, 2 μg/mL puromycin, and 100 μg/mL hygromycin B. For
transfection, HTLA cells were plated in 96-well white polystyrene
plates (Greiner Bio-One, Monroe, NC) in DMEM media supplemented
with 10% FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin at a
density of 4 × 105 cells/mL. For measuring activation of GPR37 and
GPR37L1 by TX14(A) stimulation, PDM was used instead. After 16 hr,
cells were transfected with the plasmid containing the GPR37 or
GPR37L1 ORF (Addgene, Cambridge, MA) using Trans-IT 293 (Mirus
Bio, Madison, WI) according to manufacturer's protocol. On the next
day, drugs were added in assay buffer (20 mM HEPES in HBSS,
pH 7.4) and left to incubate for 24 hr. Solutions were aspirated, and
80 μL per well of Bright-Glo solution (Promega, Madison, WI) diluted
20-fold in assay buffer was added to each well in the dark. Following
20 min of incubation, luminescence measurements were obtained
using a Tecan microplate reader (Infinite M200 PRO, Tecan Trading
AG, Männedorf, Switzerland). 2.5
| PSAP depletion To deplete PSAP from culture media (DMEM supplemented with 5%
FBS), initially, 25 μL of Protein A Magnetic Beads (NEB) per 200 μL
media were added and incubated for 1 hr at 4 C. Magnetic field was
applied for 30 s to pull beads to the side of the tube and the superna-
tant was transferred to a new tube. This step is required to remove
non-specific binding proteins. Then, 5 μL of anti-PSAP antibody
(500 μg/mL) were added to the supernatant and incubated for 1 hr at
4 C. Protein A magnetic beads were used again to pull down the
antibody-bound PSAP. Western blot was used to verify the efficient
removal of PSAP which could then be recovered from the beads
(Supporting Information Figure S4). 2.6.1
|
Luminescence activity-based GloSensor assay
(Promega) The GloSensor assay uses genetically encoded biosensor variants with
cAMP binding domains fused to mutant forms of Photinus pyralis lucif-
erase. The luminescence of the reporter increases directly proportion-
ally to the amount of cAMP present. Astrocytes were seeded in
96-well plates and transduced with an AVV bearing the CMV-driven
GLO22F at multiplicity of infection (MOI) 10. After 24 hr transduc-
tion, media was exchanged for 100 μL HEPES-buffered HBSS
(pH 7.6). Cells were then incubated with 0.731 mM beetle luciferin
for 2 hr in the dark. After baseline reading, NKH477 and/or TX14
(A) were added to the wells and incubated for 20 min. Luminescence
measurements were obtained using a Tecan microplate reader
(Infinite M200 PRO). 2.12
| Reagents Cell culture and cell-based assays related: Beetle luciferin potassium
salt (E1602, Promega); B27 (17,504,044, Life Technologies, Carlsbad,
CA); Bovine serum albumin (BSA) fraction V (A3294, Sigma, Kawasaki,
Kanagawa Prefecture, Japan); BrdU (AB142567, Abcam); Bright-Glo
reagent (E2610, Promega); cytosine β-D-arabinofuranoside (C1768,
Sigma); DNase I (D5025, Sigma); DAPI (D9542, Sigma); Dulbecco's
modified eagle medium (DMEM; 61,965, Life Technologies); fetal
bovine
serum
(FBS,
10082147,
Life
Technologies);
GlutaMax
(35,050,038, Life Technologies); L-Glutamine (2,503,008, Life Technol-
ogies); Hank's balanced salt solution (HBSS; 14175-129, Invitrogen);
HEPES (H3375, Sigma); horse serum (H1138, Sigma); hygromycin B
(H3274, Sigma); kynurenic acid (K3375, Sigma); neurobasal-A media
(10,888,022, Life Technologies); penicillin/streptomycin (15140-122,
Life Technologies); poly-D-lysine (A-003-E, Millipore); protein A mag-
netic beads (NEB, S1425S); puromycin (P8833, Sigma); Triton X-100
(T8787, Sigma); Trypsin (type III, bovine fraction; T9935, Sigma). Ambion Power SYBR Green Cells to Ct kit (4402953) was used to ver-
ify GPR37L1 and GPR37 expression in cultured and acutely isolated
astrocytes. 2.9
| DAPI staining Astrocytes were seeded in 96-well plate (1 × 104 per well) and fixed
in 4% paraformaldehyde for 5 min, washed in PBS three times for
5 min each time before and after incubating with 1 μg/mL DAPI for
10 min. Round and whole nuclei in nine fields of view per well were
counted using ImageJ (NIH, Bethesda, MD). LIU ET AL. point – Initial density of wound region)/(Density of intact cell region
at certain time point −Initial density of wound) × 100%. treated with H2O2 (250 μM, 1 hr), rotenone (50 μM, 2 hr), or staur-
osporine (100 nM, 2 hr). Stressors were then removed and neurons
were incubated with or without astrocytes inserts. After 24 hr, the
LDH reaction was carried out using 50 μL of media. Cytoprotection
was calculated using the formula: % Cytoprotection = % cytotoxic-
ity in the control condition −% cytotoxicity in the experimental
condition. 2.11
| Lactate dehydrogenase release assay Lactate dehydrogenase (LDH) release from damaged cells was
assessed colorimetrically with LDH Cytotoxicity Assay Kit (Pierce)
according to the manufacturer's instructions. Activity is proportional
to colorimetric reduction of tetrazolium salt measured at 490 nm. Cytotoxicity was normalized to maximal LDH activity as released from
cells acutely exposed to Triton X-100, and calculated using the for-
mula: % Cytotoxicity = (Compound-treated LDH activity – Spontane-
ous LDH activity)/(Maximum LDH activity – Spontaneous LDH
activity) × 100%. Antibodies: Alexa Fluor 488-conjugated goat anti-rabbit second-
ary antibody (R37116, Life Technologies); Alexa Fluor 488-conjugated
goat anti-mouse secondary antibody (R37120, Life technologies);
GPR37 L1 antibody (AB151518, Abcam, Cambridge, UK); GPR37 anti-
body (14820–1-AP, Proteintech, Chicago, IL); mouse monoclonal anti-
BrdU antibody (GTX27781, GeneTex, Irvine, CA); pan-cadherin anti-
body
(AB6529,
Abcam);
rabbit
anti-PSAP
antibody
(AB68466,
Abcam). To
determine
the
protective
effect
of
prosaptide
TX14
(A) against oxidative stress on primary astrocytes, cells were seeded
in triplicates in 96-well plates (4 × 104 per well) in PDM and trans-
duced
with
AVV–CMV–EmGFP–miR155/negative
(labeled
as
miRNA-negative in the figures), or a mixture of AVV–CMV–EmGFP–
miR155/GPR37L1 and AVV–CMV–EmGFP–miR155/GPR37 (molar
ratio 3:1, labeled as miRNA-GPR37L1/GPR37 in figures). 24 hr later,
the media was replaced by fresh PDM and cells were treated with
H2O2 (250 μM), staurosporine (200 nM), or rotenone (100 μM) for
5 hr in the presence or absence of TX14(A). The media was then
replaced by fresh PDM with or without TX14(A). Cells were incu-
bated for a further 24 hr before carrying out the LDH reaction on
50 μL of media. Drugs: 6-BenZ-cAMP (B009-10, BIOLOG Life Science Institute,
Bremen, Germany); 8-pCPT-20-O-Me-cAMP (C041-05, BIOLOG Life
Science Institute); H2O2 (H1009, Sigma); Pertussis toxin (3,097, Tocris
Bioscience, Bristol, UK); NKH 477 (SC-204130, Santa Cruz Biotech-
nology, Dallas, TX); Prosaptide TX14(A) (5,151, Tocris); rotenone
(R8875, Sigma); Staurosporine (10,042,804, Fisher Scientific, Hamp-
ton, NH). All other chemicals were from Sigma. All other chemicals were from Sigma. 2.10 To detect cell division, astrocytes were seeded in a 96-well plate
(4 × 103 per well) in media supplemented with 5% FBS or PDM and
cultured for 1 day. BrdU was added to the wells at a final concentra-
tion of 10 μM. After 24 hr, cells were fixed with 4% paraformaldehyde
in PBS (pH 7.4) for 10 min at room temperature and permeabilized
with 0.1–0.25% Triton X-100 in PBS. Cells were then incubated with
1 M HCl for 30 min, followed by primary antibody incubation with
mouse monoclonal anti-BrdU antibody (1:100) containing 2.5% goat
serum at 4 C overnight. Before imaging, cells were incubated with
Alexa Fluor 488-conjugated goat anti-mouse secondary antibody for
30 min at room temperature. 2.6.2
|
FRET-based cAMP assay Cell density was measured in the scratch area and compared to
undisrupted adjacent monolayer. Relative wound density (%), a mea-
sure of wound recovery, was calculated using the formula: Relative
wound density (%) = (Density of wound region at certain time Two to three days before recordings, astrocytes were plated onto
coverslips in prosaposin-depleted media (PDM) and transduced with
an AVV to express an Epac [Exchange protein directly activated by
cAMP)-based FRET sensor (kindly provided by K. Jalink, Amsterdam; 3.1
| GPR37L1/GPR37 activation by prosaptide
inhibits cAMP production in astrocytes but not in
HEK293 cells Body fluids, such as milk, blood, and cerebrospinal fluid, contain
PSAP (Kishimoto, Hiraiwa, & O'Brien, 1992). Most media used for
culturing are supplemented with FBS. Unsurprisingly, PSAP was pre-
sent
in
FBS-supplemented
media
(Supporting
Information
Figure S4). We studied the effect of PSAP depletion, using immu-
noadsorption (Supporting Information Figure S4), on the motility of
astrocytes in a wound scratch assay. Reproducible (700–800 μM
wide) scratch wounds were created in astrocyte monolayers. In
FBS-containing media, the wound essentially closed within 48 hr
(Figure 3a; Supporting Information Movie S1). This process was
drastically slowed down by PSAP depletion. About 100 nM TX14
(A) almost completely compensated for the loss of PSAP (Figure 3a,
c; Supporting Information Movies S2 and S3). Importantly, knock-
down of GPR37L1/GPR37 in astrocytes blocked the ability of TX14
(A) to facilitate wound closure while the control vector was ineffec-
tive (Figure 3c, Supporting Information Movies S4 and S5). Astro-
cytes divide in culture, albeit very slowly, but neither depletion of
PSAP nor addition of TX14(A) affected the number of cells in cul-
tures (Figure 3d) nor the number of newly divided cells based on
BrdU staining (Figure 3e). Therefore, the effects of PSAP and TX14
(A) in the scratch assay are due to their effect on the motility of
astrocytes. Consistent
with
published
information,
GPR37L1
is
strongly
expressed by cultured rat astrocytes which also express GPR37 at a
lower level. We have also confirmed that both receptors are present
in acutely isolated cortical astrocytes from P12 rats, consistent with
various published transcriptomes (Supporting Information Figures S2
and S3). Therefore, we always targeted both receptors simultaneously. A powerful double knock-down of GPR37L1 and GPR37 was
achieved by modifying conventional micro-RNA based cassettes to
incorporate three anti-target hairpins fused to the 30-end of the Emer-
ald green protein (Figure 1a; Liu et al., 2010). These “triple-hit” cas-
settes suppressed GPR37L1 and GPR37 protein expression below our
detection limits whereas a negative control sequence had no impact
(Figure 1b). The efficacy of the knock-down was additionally con-
firmed using real-time PCR (Supporting Information Figure S5). To
ensure maximal transduction of astrocytes we used AVV which are
exceptionally effective tools for these cells (Supporting Information
Figure S6). To assess intracellular cAMP changes, AVV were also used to
express the Glosensor biosensor (Promega). Baseline levels of cAMP
were raised ~20-fold using a water soluble forskolin analogue,
NKH477. 3.1
| GPR37L1/GPR37 activation by prosaptide
inhibits cAMP production in astrocytes but not in
HEK293 cells On that background, TX14(A) concentration dependently
decreased cAMP with an IC50 of 17.8 nM (Figure 1c). The maximal
concentration of TX14(A) used (200 nM) inhibited NKH477-mediated
cAMP production by ~40% (Figure 1c). The decrease of cAMP
induced by TX14(A) was pertussis toxin (PTX) sensitive (Figure 1d). Expression of the negative control miRNA vector had no effect while
combined knock-down of the GPR37L1 and GPR37 receptors
completely obliterated the TX14(A)-mediated decrease in cAMP levels
(Figure 1c). In addition, we employed an EPAC-based high affinity
FRET sensor for cAMP (Klarenbeek et al., 2015) to visualize cAMP
dynamics
following
TX14(A)
application
and
found
that
TX14
(A) (100 nM) significantly reduced the NKH477-evoked rise in FRET
ratio (Figure 1e,f ). Stimulation of adenylate cyclase with NKH477 (1–10 μM)
greatly slowed down wound closure, as did the cAMP analogue
6-benz-cAMP (250–1,000 μM), a selective PKA activator, in a
concentration-dependent manner (Figure 4a). The cAMP analogue
8-pCPT-20-O-Me-cAMP (250–1,000 μM) which specifically activates
EPAC (cAMP-GEF) had no effect (Figure 4a). Interestingly, nonse-
lective stimulation of cAMP production by NKH477 as well as
stimulation of PKA by 6-BenZ-cAMP and of EPAC by 8-pCPT-20-
O-Me-cAMP reduced proliferative activity of astrocytes (Figures 3d
and 4b). As all three cAMP raising drugs had a similar effect on
cell numbers but different effects on wound closure dynamics,
there appears to be no direct relationship between these two
effects of cAMP. Taken together, the data indicate that the key
mechanism of wound closure which is regulated via the GPR37L1/
GPR37 axis is the lateral movement of astrocytes, rather than their
division. Several previous reports which failed to confirm the TX14
(A) effect on GPR37L1 and GPR37 used transiently transfected
HEK293 cells. To verify that in HEK293 cells GPR37L1 and GPR37
signaling is different to that in astrocytes (Figure 1c–f ), we used the
PRESTO-Tango system (Kroeze et al., 2015). It provides GPCRs
adapted for transient expression in a HEK293 line, modified to detect
agonist-induced GPCR internalization. In that assay, GPR37 was con-
stitutively active (compared to the baseline with ADRA1; Supporting
Information Figure S7) and both receptors were insensitive to prosap-
tide TX14(A) (Figure 2), while the α-adrenoceptor 1a (ADRA1a) dem-
onstrated an appropriate response (Supporting Information Figure S7). The most likely explanation for this difference is that GPR37L1 and
GPR37 require some additional proteins for their correct function
which are lacking in HEK293 cells. 3.1
| GPR37L1/GPR37 activation by prosaptide
inhibits cAMP production in astrocytes but not in
HEK293 cells Further analysis of this issue is out-
side of the scope of the current study. 3
|
RESULTS
6 3
|
RESULTS
6 LIU ET AL. LIU ET AL. 3.2
| PSAP and TX14(A) acting via GPR37L1/GPR37
are essential for the motility of astrocytes 3 3.1
| GPR37L1/GPR37 activation by prosaptide
inhibits cAMP production in astrocytes but not in
HEK293 cells 2.13
| Statistical analysis All data analysis was performed with GraphPad Prism 7 (GraphPad
Software Inc., La Jolla, CA). One-way ANOVA with post hoc analysis
was used, unless otherwise stated. *p < .05, **p < .01, ***p < .001,
****p < .0001. Grouped data are presented as mean SD, unless oth-
erwise stated. For the protective effect of astrocytes on stressed neurons in
the co-culture system, astrocytes or neurons were transduced with
AVV at MOI 10. For astrocytes, cells were transduced at the time of
plating on cell culture inserts. For neurons, cells were transduced on
day 3 after preparation and then plated. After 24 hr, media were
replaced. Cells were cultured for 7 more days before they were Further details of statistical procedures can be found in the Sup-
porting Information as “Supplemental Statistics.” 3.3
| PSAP and prosaptide protect astrocytes from
oxidative stress damage via GPR37L1/GPR37 AVV–CMV–EmGFP–miR155/
negative is a control vector with hairpin sequence relevant to no known vertebrate gene. (c) Concentration–response curves for inhibition of
cAMP production by TX14(A) in astrocytes pretreated with 1 μM NKH477. Cells were transduced with AVV–CMV–Glosensor and either
AVV–CMV–EmGFP (control), a mixture of AVV–CMV–EmGFP–miR155/GPR37L1 and AVV–CMV–EmGFP–miR155/GPR37 to knock-down
GPR37L1/GPR37, or with AVV–CMV–EmGFP–miR155/negative (n = 4, triplicates). (d) AVV–CMV–Glosensor transduced astrocytes were
pretreated with PTX (20 hr, 100 ng/mL). About 100 nM TX14(A)-induced cAMP reduction in astrocytes was PTX sensitive (n = 12,
***p < .001 vs. indicated group, one-way ANOVA with Turkey's post hoc analysis). (e): Astrocytes expressing an EPAC-based cAMP sensor
were kept in PSAP-depleted media overnight and were stimulated with NKH477 (0.5 μM) in the absence or presence of TX14(A) (100 nM). TX14(A) decreased the transient cAMP signal; average of 58 astrocytes from four experiments. (f ) Pooled data from (e) shows significantly
decreased FRET ratio peaks with TX14(A) (n = 58 = ****p < .0001, paired t test) [Color figure can be viewed at wileyonlinelibrary.com] 3.3
| PSAP and prosaptide protect astrocytes from
oxidative stress damage via GPR37L1/GPR37 (a) Layout of the adenoviral vectors for knock-down of FIGURE 1
TX14(A) acting on GPR37L1/GPR37 reduces cAMP levels in astrocytes. (a) Layout of the adenoviral vectors for knock-down of
GPR37L1 and GPR37. Each vector allows co-cistronic expression of three pre-miRNAs targeting different regions of the target gene. FIGURE 1
TX14(A) acting on GPR37L1/GPR37 reduces cAMP levels in astrocytes. (a) Layout of the adenoviral vectors for knock-down of
GPR37L1 and GPR37. Each vector allows co-cistronic expression of three pre-miRNAs targeting different regions of the target gene. AVV = human adenoviral vectors serotype 5; CMV = human cytomegalovirus promoter; EmGFP = Emerald green fluorescent protein;
miR155 = flanking pre-miRNA sequence derived from miR-155. (b) Western blot confirms that AVV–CMV–EmGFP–miR155/GPR37L1 and
AVV–CMV–EmGFP–miR155/GPR37 (MOI 10) efficiently knock-down GPR37L1 and GPR37 in astrocytes. AVV–CMV–EmGFP–miR155/
negative is a control vector with hairpin sequence relevant to no known vertebrate gene. (c) Concentration–response curves for inhibition of
cAMP production by TX14(A) in astrocytes pretreated with 1 μM NKH477. Cells were transduced with AVV–CMV–Glosensor and either
AVV–CMV–EmGFP (control), a mixture of AVV–CMV–EmGFP–miR155/GPR37L1 and AVV–CMV–EmGFP–miR155/GPR37 to knock-down
GPR37L1/GPR37, or with AVV–CMV–EmGFP–miR155/negative (n = 4, triplicates). (d) AVV–CMV–Glosensor transduced astrocytes were
pretreated with PTX (20 hr, 100 ng/mL). About 100 nM TX14(A)-induced cAMP reduction in astrocytes was PTX sensitive (n = 12,
***p < .001 vs. indicated group, one-way ANOVA with Turkey's post hoc analysis). (e): Astrocytes expressing an EPAC-based cAMP sensor
were kept in PSAP-depleted media overnight and were stimulated with NKH477 (0.5 μM) in the absence or presence of TX14(A) (100 nM). TX14(A) decreased the transient cAMP signal; average of 58 astrocytes from four experiments. (f ) Pooled data from (e) shows significantly
decreased FRET ratio peaks with TX14(A) (n = 58 = ****p < .0001, paired t test) [Color figure can be viewed at wileyonlinelibrary.com] 14(A) acting on GPR37L1/GPR37 reduces cAMP levels in astrocytes. (a) Layout of the adenoviral vectors for kn FIGURE 1
TX14(A) acting on GPR37L1/GPR37 reduces cAMP levels in astrocytes. (a) Layout of the adenoviral vectors for knock-down of
GPR37L1 and GPR37. Each vector allows co-cistronic expression of three pre-miRNAs targeting different regions of the target gene. AVV = human adenoviral vectors serotype 5; CMV = human cytomegalovirus promoter; EmGFP = Emerald green fluorescent protein;
miR155 = flanking pre-miRNA sequence derived from miR-155. (b) Western blot confirms that AVV–CMV–EmGFP–miR155/GPR37L1 and
AVV–CMV–EmGFP–miR155/GPR37 (MOI 10) efficiently knock-down GPR37L1 and GPR37 in astrocytes. 3.3
| PSAP and prosaptide protect astrocytes from
oxidative stress damage via GPR37L1/GPR37 Exposure of astrocytes to H2O2, rotenone, or staurosporine drastically
inhibited their ability to close the wound in PSAP-depleted media but
TX14(A) rescued the stressed astrocytes' wound closure capacity
(Figure 5a). This effect of TX14(A) was eliminated by the knock-down
of GPR37L1/GPR37 in astrocytes, while the control knock-down vec-
tor was ineffective (Figure 5a). It has been previously reported that the death of cortical astro-
cytes triggered by H2O2 can be reduced by TX14(A) (Meyer et al.,
2013). We adjusted concentrations and exposure time of H2O2, rote-
none, and staurosporine to evoke a comparable degree of cytotoxicity 7 LIU ET AL. LIU ET AL. FIGURE 1
TX14(A) acting on GPR37L1/GPR37 reduces cAMP levels in astrocytes. (a) Layout of the adenoviral vectors for knock-down of
GPR37L1 and GPR37. Each vector allows co-cistronic expression of three pre-miRNAs targeting different regions of the target gene. AVV = human adenoviral vectors serotype 5; CMV = human cytomegalovirus promoter; EmGFP = Emerald green fluorescent protein;
miR155 = flanking pre-miRNA sequence derived from miR-155. (b) Western blot confirms that AVV–CMV–EmGFP–miR155/GPR37L1 and
AVV–CMV–EmGFP–miR155/GPR37 (MOI 10) efficiently knock-down GPR37L1 and GPR37 in astrocytes. AVV–CMV–EmGFP–miR155/
negative is a control vector with hairpin sequence relevant to no known vertebrate gene. (c) Concentration–response curves for inhibition o
cAMP production by TX14(A) in astrocytes pretreated with 1 μM NKH477. Cells were transduced with AVV–CMV–Glosensor and either
AVV–CMV–EmGFP (control), a mixture of AVV–CMV–EmGFP–miR155/GPR37L1 and AVV–CMV–EmGFP–miR155/GPR37 to knock-dow
GPR37L1/GPR37, or with AVV–CMV–EmGFP–miR155/negative (n = 4, triplicates). (d) AVV–CMV–Glosensor transduced astrocytes were
pretreated with PTX (20 hr, 100 ng/mL). About 100 nM TX14(A)-induced cAMP reduction in astrocytes was PTX sensitive (n = 12,
***p < .001 vs. indicated group, one-way ANOVA with Turkey's post hoc analysis). (e): Astrocytes expressing an EPAC-based cAMP sensor
were kept in PSAP-depleted media overnight and were stimulated with NKH477 (0.5 μM) in the absence or presence of TX14(A) (100 nM). TX14(A) decreased the transient cAMP signal; average of 58 astrocytes from four experiments. (f ) Pooled data from (e) shows significantly
decreased FRET ratio peaks with TX14(A) (n = 58 = ****p < .0001, paired t test) [Color figure can be viewed at wileyonlinelibrary.com]
LIU ET AL. GPR37L1/GPR37 reduces cAMP levels in astrocytes. (a) Layout of the adenoviral vectors for knock-down of FIGURE 1
TX14(A) acting on GPR37L1/GPR37 reduces cAMP levels in astrocytes. 4
|
DISCUSSION The key message of this study is that the original coupling of PSAP
and its prosaptide fragments to GPR37L1 and GPR37 suggested by
Meyer et al. (2013) was correct. This implies that we can expect small
molecules based around the structure of TX14(A) be astro- and
neuroprotective. 4.1
| Neuroprotection by PSAP PRESTO-Tango
uses clones of numerous human GPCR, C-terminally tagged with a
special signaling element. These receptors need to be expressed in a
specially designed clone of HEK293 cells. Agonist binding triggers
receptor internalization and eventually leads to expression of
luciferase and luminescence. Two concentrations of plasmid DNA
were used to express the tagged receptors. GPR37 exhibits strong
constitutive activity, especially when using 0.1 μg/μL. Values obtained
with 0.1 μg of GPR37 DNA could not be fitted with the regression
algorithm of Prism software, hence no line is shown (n = 6) [Color
figure can be viewed at wileyonlinelibrary.com] astrocytes were introduced into the wells on elevated membranes,
thereby preventing direct cell–cell contact (Figure 6a). Astrocytes
exerted a strong protective effect on the neurons which was directly
proportional to the quantity of astrocytes (Figure 6b; Supporting
Information Figure S8). About 75 k astrocytes per well provided a
near-maximum neuroprotective effect and this astrocyte density was
used for further experiments. TX14(A) applied directly to damaged cultured neurons at 100 nM,
provided a slight but significant protection against the stressors
(Figure 6c). This indicates that either neuronal GPR37L1/GPR37
receptors, or the few astrocytes remaining in the neuronal cultures,
could be contributing to the neuroprotective actions of PSAP. Figure 6d demonstrates that, for each of the three stressors, co-
culturing with astrocytes reduced LDH release by ~35–45%. Impor-
tantly, the media used in these experiments was FBS-free and con-
tained no added PSAP. However, when anti-PSAP antibodies were
added, the cytoprotective effect of astrocytes was dramatically
reduced (Figure 6d). This strongly suggests that endogenous PSAP
(or Sap C) in the co-culture system is important for the cytoprotective 3.4
| Astrocytic GPR37L1/GPR37 signaling
contributes to the protection of the cortical neurons
from damage by oxidative stressors as assessed by LDH release assay (Figure 5b). In PSAP-depleted
media, neither GPR37L1/GPR37 knock-down, nor the negative con-
trol
vector,
changed
the
cytotoxic
impact
of
stressors. TX14
(A) strongly reduced cytotoxicity which was particularly prominent in
the case of staurosporine, and this was prevented by GPR37L1/
GPR37 knock-down (Figure 5b). Neuronal cultures were subjected to the same oxidative stressors as
above and conditions were adjusted to trigger a comparable degree of
damage, based on LDH release. After removal of the stressors, LIU ET AL. LIU ET AL. 8 FIGURE 2
GPR37L1 and GPR37 are non-responsive to prosaptide
TX14(A) in PRESTO-Tango assay in HEK293 cells. PRESTO-Tango
uses clones of numerous human GPCR, C-terminally tagged with a
special signaling element. These receptors need to be expressed in a
specially designed clone of HEK293 cells. Agonist binding triggers
receptor internalization and eventually leads to expression of
luciferase and luminescence. Two concentrations of plasmid DNA
were used to express the tagged receptors. GPR37 exhibits strong
constitutive activity, especially when using 0.1 μg/μL. Values obtained
with 0.1 μg of GPR37 DNA could not be fitted with the regression
algorithm of Prism software, hence no line is shown (n = 6) [Color
figure can be viewed at wileyonlinelibrary.com] effect of astrocytes. We were able to immunodetect directly the
increase in PSAP in co-culture media in response to H2O2 stress
applied to neurons (Supporting Information Figure S9), although
the cellular source of this the stress-induced PSAP surge is cur-
rently unknown. Knock-down of GPR37L1/GPR37 selectively in
astrocytes limited astrocyte-mediated neuroprotection to a degree
comparable to PSAP depletion (Figure 6d). Interestingly, application
of the same knock-down AVV to neurons had no effect. These
results are consistent with the idea that stress-induced PSAP
release and astrocytic GPR37L1/GPR37 signaling are critical for
PSAP action. 4.1
| Neuroprotection by PSAP The first cells used to demonstrate a protective potential of PSAP
were mouse neuroblastoma NS20Y and human neuroblastoma SK-N-
MC cells (O'Brien et al., 1994). Interestingly, the neuroblastoma lines
closely related to SK-N-MC express substantial levels of GPR37
(Harenza
et
al.,
2017). Published
transcriptomes
of
astrocytes
(Anderson et al., 2016; Chai et al., 2017; Zhang et al., 2014; Zhang
et al., 2016) and our own data (Supporting Information Figures S2 and
S3) unequivocally demonstrate that astrocytes of various parts of the
central nervous system express high levels of GPR37L1 while the level
of GPR37 is generally much lower. Cytoprotective and “trophic”
effects of PSAP and its fragments were found in diverse models and
species. Prosaptides improved the outcome of sciatic nerve damage in
guinea pigs (Kotani et al., 1996), alleviated the ischemia-induced mem-
ory deficits in gerbils (Kotani et al., 1996), reduced neuropathy in dia-
betic rats (Calcutt et al., 1999), and the behavioral and anatomical
detriments caused by brain wound insult in rats (Hozumi et al., 1999). A stabilized TX14(A)-like peptide, retro–inverso prosaptide D5, was
neuroprotective in rats (Lu, Otero, Hiraiwa, & O'Brien, 2000) and ame-
liorated hyperalgesia in a model of neuropathic pain (Yan, Otero, Hir-
aiwa, & O'Brien, 2000). The neuroprotective effects of TX14(A) were
confirmed by another group (Jolivalt, Dacunha, Esch, & Calcutt, 2008;
Jolivalt, Ramos, Herbetsson, Esch, & Calcutt, 2006; Sun et al., 2002). An 18-amino acid long prosaptide was also protective in a model of
dopaminergic neuron damage (Gao et al., 2013; Sun et al., 2002). Therefore, there is solid evidence for the neuroprotective potential of
this pathway. FIGURE 2
GPR37L1 and GPR37 are non-responsive to prosaptide
TX14(A) in PRESTO-Tango assay in HEK293 cells. PRESTO-Tango
uses clones of numerous human GPCR, C-terminally tagged with a
special signaling element. These receptors need to be expressed in a
specially designed clone of HEK293 cells. Agonist binding triggers
receptor internalization and eventually leads to expression of
luciferase and luminescence. Two concentrations of plasmid DNA
were used to express the tagged receptors. GPR37 exhibits strong
constitutive activity, especially when using 0.1 μg/μL. Values obtained
with 0.1 μg of GPR37 DNA could not be fitted with the regression
algorithm of Prism software, hence no line is shown (n = 6) [Color
figure can be viewed at wileyonlinelibrary.com] FIGURE 2
GPR37L1 and GPR37 are non-responsive to prosaptide
TX14(A) in PRESTO-Tango assay in HEK293 cells. 4.2 indicated group, one-way ANOVA with Turkey's post
hoc analysis). (d): DAPI staining revealed no significant differences in the cell density between astrocytes cultured in media (+FBS), PDM and
PDM supplemented with TX14(A) (100 nM; n = 15). (e): The addition of TX14(A) to PDM does not affect the numbers of new astrocytes based
on BrdU staining (n = 7, triplicates) [Color figure can be viewed at wileyonlinelibrary.com] GPR37–mediated signaling is essential for migration of astrocytes in the scratch assay and the effect of PSAP is FIGURE 3
PSAP/GPR37L1/GPR37–mediated signaling is essential for migration of astrocytes in the scratch assay and the effect of PSAP is
mimicked by TX14(A). (a) Astrocytes move into a wound in media containing 5% FBS and in PDM (prosaposin-depleted media) supplemented
with 100 nM TX14(A) but the movement is inhibited in absence of PSAP. Representative images at 0, 24, and 48 hr post scratch. See also
Supporting Information Movies SS1–SS3. (b) Dynamics of the relative wound density under conditions shown in a (n = 3, triplicates). (c) Relative
wound density 48 hr post scratch for astrocytes incubated in FBS-containing media, PDM, PDM + TX14(A), PDM + TX14(A) + both knock-down
vectors and PDM + TX14(A) + knock-down control vector n = 6, triplicates, ***p < .001 vs. indicated group, one-way ANOVA with Turkey's post
hoc analysis). (d): DAPI staining revealed no significant differences in the cell density between astrocytes cultured in media (+FBS), PDM and
PDM supplemented with TX14(A) (100 nM; n = 15). (e): The addition of TX14(A) to PDM does not affect the numbers of new astrocytes based
on BrdU staining (n = 7, triplicates) [Color figure can be viewed at wileyonlinelibrary.com] Our findings, however, are consistent with the conclusions drawn
by (Meyer et al., 2013). The presence of PSAP in the media or addition
of TX14(A) had a powerful effect on astrocytic motility and protected
them against oxidative stressors. In all cases, this action could be
completely
prevented
by
knocking
down
GPR37L1/GPR37
(Figures 3a,c and 5). Moreover, when astrocytes were used to rescue
neurons subjected to oxidative stress, removal of GPR37L1/GPR37
only from astrocytes was sufficient to significantly weaken their neu-
roprotective capacity and block the protective action of TX14(A) (Fig-
ure 6d). 4.2 All these effects obviously called for the development of a neuropro-
tective therapy, but this opportunity could not be realized in the
absence of cognate receptors. The study by Meyer et al. (2013) l
d h
h
h
h
d
fi di
h
i
i h h
l
i
d
FIGURE 3
PSAP/GPR37L1/GPR37–mediated signaling is essential for migration of astrocytes in the scratch assay and the effect of PSAP is
mimicked by TX14(A). (a) Astrocytes move into a wound in media containing 5% FBS and in PDM (prosaposin-depleted media) supplemented
with 100 nM TX14(A) but the movement is inhibited in absence of PSAP. Representative images at 0, 24, and 48 hr post scratch. See also
Supporting Information Movies SS1–SS3. (b) Dynamics of the relative wound density under conditions shown in a (n = 3, triplicates). (c) Relative
wound density 48 hr post scratch for astrocytes incubated in FBS-containing media, PDM, PDM + TX14(A), PDM + TX14(A) + both knock-down
vectors and PDM + TX14(A) + knock-down control vector n = 6, triplicates, ***p < .001 vs. indicated group, one-way ANOVA with Turkey's post
hoc analysis). (d): DAPI staining revealed no significant differences in the cell density between astrocytes cultured in media (+FBS), PDM and
PDM supplemented with TX14(A) (100 nM; n = 15). (e): The addition of TX14(A) to PDM does not affect the numbers of new astrocytes based
on BrdU staining (n = 7, triplicates) [Color figure can be viewed at wileyonlinelibrary.com]
LIU ET AL. 9 9 LIU ET AL. 9 FIGURE 3
PSAP/GPR37L1/GPR37–mediated signaling is essential for migration of astrocytes in the scratch assa FIGURE 3
PSAP/GPR37L1/GPR37–mediated signaling is essential for migration of astrocytes in the scratch assay and the effect of PSAP is
mimicked by TX14(A). (a) Astrocytes move into a wound in media containing 5% FBS and in PDM (prosaposin-depleted media) supplemented
with 100 nM TX14(A) but the movement is inhibited in absence of PSAP. Representative images at 0, 24, and 48 hr post scratch. See also
Supporting Information Movies SS1–SS3. (b) Dynamics of the relative wound density under conditions shown in a (n = 3, triplicates). (c) Relative
wound density 48 hr post scratch for astrocytes incubated in FBS-containing media, PDM, PDM + TX14(A), PDM + TX14(A) + both knock-down
vectors and PDM + TX14(A) + knock-down control vector n = 6, triplicates, ***p < .001 vs. 10 LIU ET AL. LIU ET AL. 10 GPR37L1 is potentially indispensable for the health of the human
brain. A drastic increase in neuronal loss after an ischemic stroke in
GPR37L1 knockout mice (Jolly et al., 2017) further reinforces the
importance of these receptors. 4.3
| Coupling The first study to indicate that a Gi-coupled receptor may mediate the
action of PSAP found that pretreatment with PTX inhibited agonist-
stimulated binding of [35S]-GTPγS (Hiraiwa et al., 1997). Strikingly, it
also demonstrated that Sap C interacts with a receptor of ~54 kDa,
corresponding almost exactly to the molecular mass of GPR37L1
(Giddens et al., 2017). In our study in astrocytes, TX14(A) inhibited
cAMP production by ~40% from an elevated level set by the forskolin
analogue (Glosensor assay). The effect of TX14(A) was PTX sensitive
and the IC50 compares with that of numerous other GPCR agonists. Removal of GPR37L1 and GPR37 completely prevented the effect of
TX14(A). These results confirm the original report (Meyer et al., 2013) FIGURE 4
cAMP in astrocytes affects wound closure in the scratch
assay. (a) A scratch wound was created in astrocyte monolayers
cultured in FBS-containing media or PDM supplemented with
100 nM TX14(A). Drugs were added and dynamics of the wound
closure was monitored for 72 hr (n = 5, triplicates). (b) DAPI staining
of astrocytes shows that NKH477 (10 μM), 6-BenZ-cAMP (500 μM),
and 8-pCPT-20-O-Me-cAMP (1,000 μM) significantly decreased cell
numbers as compared to control (n = 4, triplicates). In both cases, one-
way ANOVA with Dunnett's post hoc analysis. **p < .01, ***p < .001
vs. control [Color figure can be viewed at wileyonlinelibrary.com] astrocytes (Figure 6c). At least, application of the knock-down strat-
egy to neurons was without an obvious effect (Figure 6d). Therefore, by several approaches, we demonstrate that the
effects of PSAP and TX14(A) on astrocytes are invariably dependent
on GPR37L1/GPR37. The partial protection provided to injured neu-
rons by co-cultured astrocytes to some extent also depends on this
signaling pathway. Given that only removal of GPR37L1/GPR37 from
astrocytes, but not neurons interfered with neuroprotection in this
paradigm, the likeliest scenario is that PSAP acts as an autocrine signal
on the receptors located on the astrocytes to recruit additional, uni-
dentified, neuroprotective molecules (Figure 7). FIGURE 5
TX14(A) acts via GPR37L1 and GPR37 to protect primary
astrocytes against toxicity induced by H2O2, staurosporine or
rotenone. (a) Pre-exposure to stressors (5 hr) drastically reduce
relative wound density recorded at 48 hr in PDM. TX14(A) (100 nM)
rescued astrocytes bringing wound density close to normal (compare
to Figure 3). 4.2 TX14(A) at a fairly high concentration (100 nM) had a weak
direct protective effect on stressed neurons which is unlikely to make
a major contribution to the neuroprotection seen in the presence of strongly suggested that these are the orphan receptors GPR37L1 and
GPR37. Some of the effects were demonstrated in HEK293 cells, but
the most striking protective effect of TX14(A) was observed in cul-
tured astrocytes. The findings of Meyer et al. (2013) were later criti-
cized, the key concerns being the high concentration of TX14(A) used,
the small magnitude of the Gi-mediated inhibitory effect on cAMP
concentration and the failure to detect TX14(A) agonism in a
β-arrestin-based DiscoverX assay (Smith, 2015; Southern et al., 2013). Other recent studies reported high constitutive activity of GPR37L1
and GPR37 and lack of a TX14(A) effect in either HEK293 cells or
yeast (Coleman et al., 2016; Giddens et al., 2017; Ngo et al., 2017;
Southern et al., 2013). astrocytes (Figure 6c). At least, application of the knock-down strat-
FIGURE 4
cAMP in astrocytes affects wound closure in the scratch
assay. (a) A scratch wound was created in astrocyte monolayers
cultured in FBS-containing media or PDM supplemented with
100 nM TX14(A). Drugs were added and dynamics of the wound
closure was monitored for 72 hr (n = 5, triplicates). (b) DAPI staining
of astrocytes shows that NKH477 (10 μM), 6-BenZ-cAMP (500 μM),
and 8-pCPT-20-O-Me-cAMP (1,000 μM) significantly decreased cell
numbers as compared to control (n = 4, triplicates). In both cases, one-
way ANOVA with Dunnett's post hoc analysis. **p < .01, ***p < .001
vs. control [Color figure can be viewed at wileyonlinelibrary.com]
10 4.4
| Controversy related to PSAP-GPR37L1/GPR37
pairing Neither HEK293 cells nor yeast used in the recent conflicting studies
(Coleman et al., 2016; Giddens et al., 2017; Ngo et al., 2017) express
GPR37L1 or GPR37 natively, nor were they ever demonstrated to be
responsive to PSAP or prosaptides. These cells might not necessarily
recapitulate the intracellular environment of astrocytes and neurons
which are the natural habitats of GPR37L1 and GPR37. Our experi-
ments using PRESTO-Tango confirmed that neither GPR37L1 nor
GPR37 respond to TX14(A) in HEK293 cells (Figure 2). Serum-containing media can mask the effects of GPR37L1/
GPR37 ligands because it contains considerable levels of PSAP. Most
likely astrocytes, neurons or both can secrete some PSAP in vivo. This
was visible in the “rescue” experiments where astrocytes reduced the
damage caused to neurons by added stressors. In these experiments,
anti-PSAP antibodies reduced neuroprotection even though the media
was nominally devoid of PSAP. In the presence of stressed neurons,
PSAP was greatly upregulated and easily detected in the media by
immunoblotting. Therefore, to fully reveal the agonist activity of TX14
(A), it is important to ensure that the receptors are not persistently
exposed to endogenous or media-derived PSAP. FIGURE 6
Co-cultured astrocytes protect cortical neurons against
oxidative toxicity partially through GPR37L1/GPR37 signaling in
astrocytes. (a) Experimental design: (A—stressed neurons (blue) in
absence or presence of astrocytes (green) on a culture insert; B—
depletion of PSAP from the media with anti-PSAP antibodies; C—
GPR37L1 and GPR37 knock-down in astrocytes co-cultured with
neurons; D—GPR37L1 and GPR37 knock-down in neurons in the co-
culture system. Neurons were treated with H2O2 (250 μM) for 1 hr, or
staurosporine (100 nM) or rotenone (50 μM) for 2 hr. Stressors were
then removed and inserts with astrocytes were introduced. LDH
assay was carried out 24 hr later. (b) Astrocytes protect cortical
neurons against H2O2-induced stress, the effect saturates at ~75 k
astrocytes per co-culture (n = 5, triplicates, ***p < .001 vs. control
[no astrocytes insert], ****p < .0001 vs. groups with less than 50 k
astrocytes, one-way ANOVA with Tukey's post hoc analysis). (c) TX14
(A) has a weak protective effect on neurons against oxidative stress
(n = 6, triplicates, ***p < .001, ****p < .0001 vs. 50 nM TX14
(A) group (effect of 50 nM is not significant, one-way ANOVA with
Bonferoni's post hoc analysis). LIU ET AL. 11 FIGURE 6
Co-cultured astrocytes protect cortical neurons against
oxidative toxicity partially through GPR37L1/GPR37 signaling in
astrocytes (a) Experimental design: (A stressed neurons (blue) in prevented by knock-down of the two receptors (Figure 3c). Interestingly,
stimulants of the cAMP pathway, the adenylyl cyclase activator
NKH477,
and
two
pathway-biased
agonists,
6-BenZ-cAMP,
and
8-pCPT-20-O-Me-cAMP, reduced the speed of division of cultured
astrocytes (Figure 4b) but only 6-BenZ-cAMP which predominantly acti-
vates PKA (Bos, 2003) concentration-dependently suppressed spread of
the astrocytes into the wounded area (Figure 4a). Therefore, the motility
of astrocytes is regulated by PKA, rather than EPAC-regulated proteins. TX14(A) did not interfere with the reduction in mitotic activity by either
of the cAMP analogs (Figure 4b). The mechanism by which PKA regu-
lates astrocyte motility is currently unknown. 4.3
| Coupling GPR37L1/GPR37 knock-down prevented the protective
effect of TX14(A), while the control vector had no effect (n = 6,
triplicates, ***p < .001 vs. indicated group). (b) LDH release was used
as a measure of cytotoxicity, 24 hr after exposure of astrocytes to
oxidative stress. In PDM, manipulation of GPR37L1/GPR37 had no
effect. TX14(A) (100 nM) protected them from damage but only when
they were expressing GPR37L1/GPR37, (n = 6, triplicates, **p < .01,
***p < .001 vs. control group, for example, PDM groups). One-way
ANOVA with Bonferroni's post hoc analysis [Color figure can be
viewed at wileyonlinelibrary.com] FIGURE 5
TX14(A) acts via GPR37L1 and GPR37 to protect primary
astrocytes against toxicity induced by H2O2, staurosporine or
rotenone. (a) Pre-exposure to stressors (5 hr) drastically reduce
relative wound density recorded at 48 hr in PDM. TX14(A) (100 nM)
rescued astrocytes bringing wound density close to normal (compare
to Figure 3). GPR37L1/GPR37 knock-down prevented the protective
effect of TX14(A), while the control vector had no effect (n = 6,
triplicates, ***p < .001 vs. indicated group). (b) LDH release was used
as a measure of cytotoxicity, 24 hr after exposure of astrocytes to
oxidative stress. In PDM, manipulation of GPR37L1/GPR37 had no
effect. TX14(A) (100 nM) protected them from damage but only when
they were expressing GPR37L1/GPR37, (n = 6, triplicates, **p < .01,
***p < .001 vs. control group, for example, PDM groups). One-way
ANOVA with Bonferroni's post hoc analysis [Color figure can be
viewed at wileyonlinelibrary.com] Previous studies in mice lacking either of the two receptors dem-
onstrated various, albeit relatively mild, phenotypes but, to the best of
our knowledge, the possibility of compensation by the remaining
receptor in a single knockout scenario has never been explored. This
could be the reason why the pro-seizure phenotype of double
GPR37L1/GPR37 knockout mice is so severe (Giddens et al., 2017). Recent demonstration of a lethal neurological phenotype in humans
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variant. Neurobiology of Disease, 106, 181–190. https://doi.org/10. 1016/j.nbd.2017.07.006 ACKNOWLEDGMENTS This study was supported by the grants from MRC, MR/L020661/1
and BBSRC, BB/L019396/1. B. V. C. is funded by a Science without
Borders scholarship from the CNPq (206427/2014-0). The authors
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setting up Glosensor assay. Goldman, J. E., & Chiu, F. C. (1984). Dibutyryl cyclic AMP causes interme-
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| Controversy related to PSAP-GPR37L1/GPR37
pairing D: PSAP depletion or GPR37L1/
GPR37 knock-down selectively in astrocytes significantly attenuates
the protective effect of astrocytes on neurons pre-exposed to
oxidative stressors (n = 6, triplicates, ***p < .001 vs. indicated group,
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Working model of the neuroprotective role of astrocytic
GPR37L1/GPR37 based on the evidence presented in this study. Damaged neurons release diffusible “SOS” factor(s) which trigger
release of PSAP. PSAP acts on GPR37L1 on astrocytes and activates
release of diffusible neuroprotective factor(s) [Color figure can be
viewed at wileyonlinelibrary.com] FIGURE 7
Working model of the neuroprotective role of astrocytic
GPR37L1/GPR37 based on the evidence presented in this study. Damaged neurons release diffusible “SOS” factor(s) which trigger
release of PSAP. PSAP acts on GPR37L1 on astrocytes and activates
release of diffusible neuroprotective factor(s) [Color figure can be
viewed at wileyonlinelibrary.com] that GPR37L1/GPR37 are Gi-coupled receptors. Coupling to other G-
proteins needs to be investigated further. In the wound scratch assay, PSAP and TX14(A) were permissive to
the spread of astrocytes into the barren area. This was due to an effect
on lateral motility (Figure 3d,e). Again, this effect was completely LIU ET AL. 13 residues of the prosaposin sequence has neurotrophic activity in vitro
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mediated by orphan G-protein coupled receptors GPR37L1
and GPR37. Glia. 2018;1–13. https://doi.org/10.1002/glia. 23480 O'Brien, J. S., Carson, G. S., Seo, H. C., Hiraiwa, M., & Kishimoto, Y. (1994). Identification of prosaposin as a neurotrophic factor. Proceedings of the
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Arthroskopie
AGA-Studenten
Arthroskopie 2021 · 34:70–73
https://doi.org/10.1007/s00142-020-00424-9
Angenommen: 26. November 2020
Online publiziert: 16. Dezember 2020
© Der/die Autor(en) 2020
Lars Brunnader1 · Stefan Greiner2,3 · Andreas Voss2,3
1
Medizinische Universität Graz, Graz, Österreich
sporthopaedicum, Regensburg/Straubing, Deutschland
3
Klinik und Poliklinik für Unfallchirurgie, Universitätsklinik Regensburg, Regensburg, Deutschland
2
Superiore Kapselrekonstruktion
mittels langer Bizepssehne bei
retrahierter Komplettruptur der
Supraspinatussehne und der
kranialen Infraspinatussehne
Die Rotatorenmanschette besteht aus den
Sehnen folgender Muskeln: M. infraspinatus (ISP), M. supraspinatus (SSP),
M. subscapularis und M. teres minor,
welche alle von der Skapula entspringen und am Humeruskopf ansetzen.
Das Lig. korakohumerale vervollständigt die Rotatorenmanschette [1], deren Hauptfunktion darin besteht, den
Humeruskopf im Zentrum der Fossa
glenoidale zu halten, um somit sowohl
die Abduktion als auch die Anteversion des Arms effizient zu gestalten [2].
Rupturen der Rotatorenmanschette präsentieren sich hinsichtlich Pathogenese,
der Rupturmorphologie und den daraus
resultierenden funktionellen Einschränkungen in variablen Ausprägungen. Die
am häufigsten beschriebene Läsion ist
die posterosuperiore Rotatorenmanschettenruptur. Diese ist in den meisten
Fällen degenerativ bedingt. Betroffen
sind meist die Supraspinatussehne, der
kraniale Anteil der Infraspinatussehne
und der komplette Footprint der Infraspinatussehne [3]. Die Einteilung der
Rotatorenmanschettenruptur nach Bateman richtet sich nach der Rupturgröße:
4 Grad 1: Rupturgröße ≤1 cm,
4 Grad 2: Rupturgröße 1–3 cm,
4 Grad 3: Rupturgröße 3–5 cm,
4 Grad 4: Rupturgröße ≥5 cm.
Die Sehnenretraktion kann nach Patte
eingeteilt werden. Bei Grad 1 befindet
sich der Sehnenstumpf zwischen Tuber-
70
Arthroskopie 1 · 2021
culum majus und dem Apex des Humeruskopfes, bei Grad 2 liegt der Stumpf
zwischen dem Apex und dem Glenoidrand und bei Grad 3 hinter dem Glenoidrand [4].
Fallbeschreibung
Der 57-jährige Patient stellte sich mit
seit 6 Monaten bestehenden Schmerzen in der linken Schulter vor. Die
Schmerzsymptomatik konnte keinem
Trauma zugeordnet werden. Eine einmalig e. d. durchgeführte Injektionstherapie (Medikament bei Vorstellung
unbekannt) führte zu einer leichten Verbesserung der Beschwerdesymptomatik.
Neben Schmerzen, die insbesondere bei
Überkopfbewegungen und Belastung
bestanden, konnte auch ein Kraftverlust
detektiert werden.
Bei der körperlichen Untersuchung
ließsicheine freie aktive sowie passive Beweglichkeit feststellen. Der Nackengriff
war vollständig durchführbar, der Schürzengriff konnte bis zum 1. Lendenwirbelkörper ausgeführt werden, Bear-Hug
und Speed-Test waren positiv. Die Kraftprüfung nach Janda zeigte eine Kraftminderung des M. supraspinatus (Janda
Stufe 3) und des M. infraspinatus (Janda
Stufe 4), bei der Testung des M. subscapularis konnte hingegen keine Kraftminderung festgestellt werden.
In der Magnetresonanztomographie
(MRT; . Abb. 1) ließ sich eine Komplett-
ruptur der Supraspinatussehne und der
kranialen Infraspinatussehne objektivieren. Die Rissgröße wurde mit Grad 3 nach
Bateman klassifiziert, die Retraktion des
Sehnenstumpfes wurde als Grad 3 nach
Patte und die trophische Muskelqualität als Grad 3 nach Goutallier eingestuft.
Darüber hinaus zeigte sich eine Tendinitis der langen Bizepssehne (LBS). In
der Röntgenbefundung der linken Schulter präsentierten sich ein Humeruskopfhochstand und eine Arthrose des Akromioklavikulargelenks.
In Zusammenschau der Befunde war
eine Weiterführung der konservativen
Therapie aus der Sicht des behandelnden
Arztes nicht erfolgversprechend, weshalb
man sich gemeinsam mit dem Patienten
zur operativen Therapie in Form einer
Schulterarthroskopie entschloss, in welcher in erster Linie eine Rekonstruktion
der superioren Kapsel mittels LBS-Patch
und Readaptation des Infraspinatus, eine
Tenotomie der langen Bizepssehne (LBS)
und eine subakromiale Dekompression
durchgeführt werden sollten.
Der primäre arthroskopische Zugang
erfolgte über das posteriore Standardportal, anschließend wurde das anterosuperiore Arbeitsportal gesetzt. Bei der
palpatorischen und dynamischen Untersuchung zeigte sich das Pulley-System
der Bizepssehne rupturiert. Der radiologische Befund der Supra- und Infraspinatussehne bestätigte sich auch arthroskopisch, des Weiteren war eine deutli-
Abb. 1 8 Magnetresonanztomographie (MRT) der Schulter. In der T2-gewichteten koronaren Aufnahme (a) ist die Komplettruptur der Supraspinatussehne deutlich zu sehen, während sich in der axialen
T1-gewichteten Aufnahme (b) die Ruptur der kranialen Infraspinatussehne objektivieren lässt
che Bursitis zu sehen. Anschließend wurde über den posterioren Zugang in den
Subakromialraum eingegangen, woraufhin sich auch hier eine deutliche Bursitis zeigte. Anschließend wurden mittels Shaver eine Bursektomie der Bursa
subacromialis und eine elektrothermische Blutstillung durchgeführt. Durchdie
Präzisionsakromioplastik nach Ellmann,
welche mithilfe einer rotierenden Fräse
umgesetzt wurde, konnte eine gute subakromiale Defilee-Erweiterung erreicht
werden. Nach dem erneuten Eingehen
in den Gelenkraum erfolgte auch hier
zunächst die Bursektomie mit anschließender Blutstillung.
Im nächsten Schritt wurde ein laterales Arbeitsportal gesetzt, wodurch
eine Twist-in-Arbeitskanüle (Arthrex
GmbH, München, Deutschland) eingebracht wurde. Die folgende Anfrischung
des SSP- und ISP-Footprint wurde mit
dem Shaver durchgeführt. Hier wurde
darauf geachtet, dass dies solange erfolgte bis Blutaustritte aus dem Knochen
zu sehen waren. Auch der superiore
Glenoidrand wurde mit dem Shaver
präpariert.
Danach wurde die lange Bizepssehne armiert und beim Eintritt in
den Sulcus intertubercularis mit zwei
FiberWire -Fäden (Arthrex GmbH,
München, Deutschland) mittels einer
arthroskopischen FastPass Scorpion
SL-Zange (Arthrex GmbH, München,
Deutschland) durchstochen, woraufhin
die lange Bizepssehne distal der Armierung, im Sinne einer Tenotomie,
abgesetzt und somit als Autograft für
die obere Kapsel vorbereitet wurde. Der
nächste Schritt bestand in der Fixierung der langen Bizepssehne mit zwei
5,5-mm-SwiveLock -Ankern (Arthrex
GmbH, München, Deutschland), jeweils am ventralen und dorsalen Rand
(. Abb. 2).
Mit dem freien FibreWire -Faden
wurde die Infraspinatussehne durchstochen und so eine Teilrekonstruktion
des ISP durchgeführt. Danach erfolgte
die Side-to-side-Naht der langen Bizepssehne durch Verknüpfung mit der
kranialen Infraspinatussehne, um so den
kompletten Verschluss des posterosuperioren Gelenkraums zu bewerkstelligen
(. Abb. 3).
®
Abb. 2 8 Abgesetzte lange Bizepssehne (LBS)mit ventralem (V)unddorsalem (D)Rand(a),Einbringen
eines Fadenankers im Supraspinatus(SSP)-Footprint (b)
®
®
Abb. 3 8 Durstechen (a) von langer Bizepssehne (LBS) und Infraspinatussehne (ISP) zur Komplettierung (b) des Verschlusses des posterosuperioren Gelenkraums
Arthroskopie 1 · 2021
71
Zusammenfassung · Abstract
Siebeneinhalb Monate nach dem Eingriff war jeweils eine schmerzfreie aktive Abduktion und Flexion bis 90° möglich. Der Schürzengriff konnte bis zum
5. Lendenwirbelkörper ausgeführt werden, während der Nackengriff mit vorgeneigtem Ellenbogen durchgeführt werden konnte.
Diskussion
Pathologien der Rotatorenmanschette sind für ca. 70 % aller ärztlichen
Konsultationen im Rahmen von Schulterschmerzen verantwortlich [5]. Die
Inzidenz der Rotatorenmanschettenrupturen steigt mit dem Alter und beträgt
bei Personen zwischen dem 60. und
70. Lebensjahr etwa 25 %, über dem
80. Lebensjahr hingegen schon über
50 % [6]. Ein Defekt der superioren
Kapsel hat biomechanisch eine erhöhte
glenohumerale Translation in alle Richtungen zur Folge [7]. Der Verlust dieses
stabilisierenden Faktors des Glenohumeralgelenks führt dazu, dass v. a. eine
durch den M. deltoideus bedingte Translation nach anterosuperior geschieht, was
zu einer unzureichenden Funktion der
Schulter führt.
Ziel dersuperiorenKapselrekonstruktion (SCR) ist eine Verbesserung der oben
beschriebenen Problematik. Durch eine
Rezentrierung des Humeruskopfes kann
die ursprüngliche Biomechanik wiederhergestellt werden. Mihata et al. leisteten
dahingehend Pionierarbeit und bewiesen, dass durch eine SCR die superiore
Stabilität der Rotatorenmanschette wieder vollständig hergestellt werden kann
[8].
Ein Fascia-lata-Allotransplan»tat sollte
zumindest 8 mm Dicke
aufweisen
Die ursprüngliche SCR-Technik wurde
von Mihata et al. in Form eines Fascia-lata-Allotransplantats beschrieben, später
in Form eines Fascia-lata-Autotransplantats [8]. Aus Überlegungen der kürzeren Operationszeit, der verringerten Invasivität betreffend der Entnahme und
der einfacheren Herstellung gewann das
Allograft immer mehr an Popularität,
72
Arthroskopie 1 · 2021
obwohl die Gleichwertigkeit der Transplantate noch nicht nachgewiesen ist [9].
Sowohl Fascia-lata-Allotransplantate als
auch humane dermale Allotransplantate
scheinen in der Lage zu sein, die Humeruskopftranslation zu verringern [10].
Die Dicke des gewählten Transplantats ist
ein entscheidender Faktor, inwiefern die
Biomechanik des Glenohumeralgelenks
wiederhergestellt werden kann. So sollte
ein Fascia-lata-Allotransplantat zumindest 8 mm Dicke aufweisen, sodass es die
Translation zum Großteil beheben kann
[11].
Die im Fallbericht beschriebene Technik der Verwendung der LBS zur Rekonstruktion der superioren Kapsel hat
gegenüber der Verwendung eines Fascialata-Autotransplantats den Vorteil, sich
einen weiteren Einschnitt zu ersparen
und so möglichen Komplikationen, wie
z. B. Narben und Hämatombildung, von
vorneherein vorzubeugen. Auch diese
Technik ist in der Lage, den Humeruskopf zu rezentrieren. Darüber hinaus
werden der subakromiale Spitzendruck
und die Kontaktfläche von Humerus und
Glenoid signifikant verringert [12, 13].
Ebenso konnte bewiesen werden, dass
auch das Fortschreiten von Arthrose
oder die Atrophie der Muskulatur der
Rotatorenmanschette durch dieses Verfahren verlangsamt werden kann [14].
In mehreren Studien konnte zudem eine
signifikante Verbesserung des funktionellen Outcomes sowie eine signifikante
Abnahme der Schmerzen nachgewiesen
werden [15]. Was die lange Bizepssehne
als Transplantat für superiore Kapseldefekte zudem noch interessanter macht,
ist der Umstand, dass sie eine geeignete Zellquelle für die Regeneration von
Sehnengewebe darstellt [16].
Da es bei einigen Patienten neben dem
Ruptur der Rotatorenmanschette jedoch
auch zu einer Degeneration und Rissbildung der langen Bizepssehne kommt
[15], ist dieses Verfahren nicht immer
geeignet.
Fazit für die Praxis
4 In dem hier beschriebenen Fall
konnte die Rotatorenmanschettenruptur eines 57-jährigen Patienten
Arthroskopie 2021 · 34:70–73
https://doi.org/10.1007/s00142-020-00424-9
© Der/die Autor(en) 2020
L. Brunnader · S. Greiner · A. Voss
Superiore
Kapselrekonstruktion mittels
langer Bizepssehne bei
retrahierter Komplettruptur
der Supraspinatussehne und
der kranialen
Infraspinatussehne
Zusammenfassung
In diesem Beitrag berichten die Autoren
über einen Patienten mit einer Ruptur
der Rotatorenmanschette, welche sich
als retrahierte Komplettruptur der
Supraspinatussehne und der kranialen
Infraspinatussehne präsentierte. Nach
7-monatiger Schmerzpersistenz und
anhaltendem Funktionsverlust wurde im
Rahmen eines arthroskopischen Eingriffs
eine Rekonstruktion der superioren Kapsel
durchgeführt. Die lange Bizepssehne wurde
hierzu als Autograft verwendet und mit der
kranialen Infraspinatussehne verknüpft.
Auf diese Weise konnte ein kompletter
Verschluss des posterosuperioren
Gelenkraums erreicht werden.
Schlüsselwörter
Rotatorenmanschettenruptur · Supraspinatussehne · Infraspinatussehne · Transplantat ·
Magnetresonanztomographie
Superior capsule
reconstruction using the long
biceps tendon for retracted
complete rupture of the
supraspinatus tendon and the
cranial infraspinatus tendon
Abstract
This article reports the case of a patient
with a massive rupture of the rotator cuff,
which presented as a retracted complete
rupture of the supraspinatus tendon and
the cranial infraspinatus tendon. After 7
months of persisting pain and continued
loss of function, the superior capsule was
reconstructed within the framework of an
arthroscopic intervention. The long biceps
tendon was used as an autograft and linked
to the cranial infraspinatus tendon. In this
way it was possible to achieve a complete
closure of the posterosuperior joint space.
Keywords
Rotator cuff tear · Supraspinatus tendon ·
Infraspinatus tendon · Autograft · Magnetic
resonance imaging
erfolgreich mittels superiorer Kapselrekonstruktion behandelt werden.
4 Hierzu wurde die lange Bizepssehne
als Autograft verwendet und mit
der kranialen Infraspinatussehne
verknüpft.
4 Mittels dieser Technik konnten Biomechanik und Stabilität des Schultergelenks konnten wiederhergestellt
werden.
4 Die lange Bizepssehne eignet sich,
sofern sie keine Degeneration oder
Rissbildung aufweist, aufgrund ihrer
Eigenschaften prinzipiell sehr gut als
Transplantat.
Korrespondenzadresse
Lars Brunnader
Medizinische Universität Graz
Graz, Österreich
lars.brunnader@gmail.com
Funding. Open access funding provided by Medical
University of Graz.
Einhaltung ethischer Richtlinien
Interessenkonflikt. L. Brunnader, S. Greiner und
A. Voss geben an, dass kein Interessenkonflikt besteht.
Für diesen Beitrag wurden von den Autoren keine
Studien an Menschen oder Tieren durchgeführt. Für
die aufgeführten Studien gelten die jeweils dort angegebenen ethischen Richtlinien. Für Bildmaterial oder
anderweitige Angaben innerhalb des Manuskripts,
über die Patienten zu identifizieren sind, liegt von
ihnen und/oder ihren gesetzlichen Vertretern eine
schriftliche Einwilligung vor.
Open Access. Dieser Artikel wird unter der Creative
Commons Namensnennung 4.0 International Lizenz
veröffentlicht, welche die Nutzung, Vervielfältigung,
Bearbeitung, Verbreitung und Wiedergabe in jeglichem Medium und Format erlaubt, sofern Sie den/die
ursprünglichen Autor(en) und die Quelle ordnungsgemäß nennen, einen Link zur Creative Commons Lizenz
beifügen und angeben, ob Änderungen vorgenommen wurden.
Die in diesem Artikel enthaltenen Bilder und sonstiges
Drittmaterial unterliegen ebenfalls der genannten
Creative Commons Lizenz, sofern sich aus der Abbildungslegende nichts anderes ergibt. Sofern das betreffende Material nicht unter der genannten Creative
Commons Lizenz steht und die betreffende Handlung
nicht nach gesetzlichen Vorschriften erlaubt ist, ist für
die oben aufgeführten Weiterverwendungen des Materials die Einwilligung des jeweiligen Rechteinhabers
einzuholen.
Weitere Details zur Lizenz entnehmen Sie bitte der
Lizenzinformation auf http://creativecommons.org/
licenses/by/4.0/deed.de.
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73.
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https://openalex.org/W2580085487
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OpenAlex
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Open Science
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CC-By
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Diagnostic performance of imaging modalities in chronic pancreatitis: a systematic review and meta-analysis
|
Yama Issa
|
English
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Spoken
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Abstract Objectives Obtain summary estimates of sensitivity and spec-
ificity for imaging modalities for chronic pancreatitis (CP)
assessment. • EUS, ERCP, MRI and CT have high diagnostic sensitivity for
chronic pancreatitis Methods A systematic search was performed in Cochrane
Library, MEDLINE, Embase and CINAHL databases for
studies evaluating imaging modalities for the diagnosis of
CP up to September 2016. A bivariate random-effects model-
ing was used to obtain summary estimates of sensitivity and
specificity. • Diagnostic specificity is comparable for all imaging
modalities • EUS and ERCP are outperformers and US has the lowest
accuracy • The choice of imaging can be made based on clinical
considerations • The choice of imaging can be made based on clinical
considerations Results We included 43 studies evaluating 3460 patients. Sensitivity of endoscopic retrograde cholangiopancreatography
(ERCP) (82%; 95%CI: 76%-87%) was significant higher than
that of abdominal ultrasonography (US) (67%; 95%CI: 53%-
78%; P=0.018). The sensitivity estimates of endoscopic ultraso-
nography (EUS), magnetic resonance imaging (MRI), and com-
puted tomography (CT) were 81% (95%CI: 70%-89%), 78%
(95%CI: 69%-85%), and 75% (95%CI: 66%-83%), respective-
ly, and did not differ significantly from each other. Estimates of
specificity were comparable for EUS (90%; 95%CI: 82%-95%),
ERCP (94%; 95%CI: 87%-98%), CT (91%; 95% CI: 81%-
96%), MRI (96%; 95%CI: 90%-98%), and US (98%; 95%CI:
89%-100%). Results We included 43 studies evaluating 3460 patients. Sensitivity of endoscopic retrograde cholangiopancreatography
(ERCP) (82%; 95%CI: 76%-87%) was significant higher than
that of abdominal ultrasonography (US) (67%; 95%CI: 53%-
78%; P=0.018). The sensitivity estimates of endoscopic ultraso-
nography (EUS), magnetic resonance imaging (MRI), and com-
puted tomography (CT) were 81% (95%CI: 70%-89%), 78%
(95%CI: 69%-85%), and 75% (95%CI: 66%-83%), respective- Keywords Chronic pancreatitis . Diagnostic imaging . Diagnostic accuracy . Meta-analysis Diagnostic accuracy . Meta-analysis Eur Radiol (2017) 27:3820–3844
DOI 10.1007/s00330-016-4720-9 GASTROINTESTINAL Diagnostic performance of imaging modalities in chronic
pancreatitis: a systematic review and meta-analysis Y. Issa1 & M. A. Kempeneers1 & H. C. van Santvoort1 & T. L. Bollen2 & S. Bipat3 &
M. A. Boermeester1 Y. Issa1 & M. A. Kempeneers1 & H. C. van Santvoort1 & T. L. Bollen2 & S. Bipat3 &
1 Received: 3 May 2016 /Revised: 20 September 2016 /Accepted: 16 December 2016 /Published online: 27 January 2017
# The Author(s) 2017. This article is published with open access at Springerlink.com and ERCP are outperformers and US has the lowest accuracy. The choice of imaging modality can therefore be made based
on invasiveness, local availability, experience and costs. Key Points and ERCP are outperformers and US has the lowest accuracy. The choice of imaging modality can therefore be made based
on invasiveness, local availability, experience and costs. Key Points 1
Department of Surgery, Academic Medical Centre, Meibergdreef 9,
1100DD Amsterdam, The Netherlands * M. A. Boermeester
m.a.boermeester@amc.uva.nl 2
Department of Radiology, St Antonius Ziekenhuis, Koekoekslaan 1,
3430EM Nieuwegein, The Netherlands 3
Department of Radiology, Academic Medical Centre, Meibergdreef
9, 1100DD Amsterdam, The Netherlands Introduction Chronic pancreatitis (CP) is a disabling inflammatory disease
of the pancreas characterized by severe recurrent or continu-
ous abdominal pain and considerable impact on the quality of
life [1–4]. Patients with CP usually develop endocrine and
exocrine insufficiency during the course of the disease as a
result of the progressive loss of pancreatic parenchyma. Conclusions EUS, ERCP, MRI and CT all have comparable
high diagnostic accuracy in the initial diagnosis of CP. EUS There is lack of international consensus regarding the ini-
tial diagnosis of CP, particularly at its early stages. The diag-
nosis is often made by a combination of clinical symptoms
(e.g. abdominal pain, malabsorption, diabetes mellitus), pan-
creatic function tests (e.g. fecal elastase-1) and morphological
abnormalities seen on imaging (e.g. calcifications, ductal le-
sions, pseudocysts) [5, 6]. Imaging plays a key role in the
diagnosis and therapeutic management of patients with CP. The most frequently used imaging modalities for CP are en-
doscopic ultrasonography (EUS), endoscopic retrograde Eur Radiol (2017) 27:3820–3844 3821 cholangiopancreatography (ERCP), magnetic resonance im-
aging (MRI), computed tomography (CT) and ultrasonogra-
phy (US). Data was extracted regarding the imaging characteristics:
type of imaging modality, scoring criteria, technical features
for each modality, and reported observer experience. Also data
on the reference standard was extracted, such as clinical fol-
low-up, surgery and histology. cholangiopancreatography (ERCP), magnetic resonance im-
aging (MRI), computed tomography (CT) and ultrasonogra-
phy (US). The aim of this meta-analysis was to determine the diag-
nostic accuracy of imaging modalities for the initial diagnostic
assessment of CP. The methodological quality of the included articles
was assessed by the Quality Assessment of Diagnostic
Accuracy Studies version 2 (QUADAS-2) tool [7]. The
QUADAS-2 tool evaluates the risk of bias in four do-
mains (patient selection, index test, reference standard,
flow and timing) and the clinical applicability in the
first three domains. Signaling questions were used to
help assess the risk of bias and applicability. Possible
answers were ‘yes’, ‘no’ or ‘unclear’ in which ‘yes’
indicates no risk of bias. In addition the GRADE scor-
ing system for diagnostic tests was used, which assesses
the quality of evidence for each imaging modality [8,
9]. Although the criteria are applicable to diagnostic test
accuracy, the methods are less well established com-
pared to interventional studies [10]. Search A search was performed in Cochrane Library, MEDLINE,
EMBASE and CINAHL databases, without restrictions for
publication date or language up to September 2016. The
search included terms for chronic pancreatitis, EUS, ERCP,
MR imaging, CT and US. For detailed search details, see
Appendix Table 5. Selection of studies All search hits were screened on title and abstract and eligible
articles on full text by two reviewers independently (YI and
MAK). Disagreements were solved through discussion with a
third reviewer (MAB). Studies were eligible when EUS,
ERCP, MR imaging, CT or US was evaluated in patients with
suspected CP. Duplicates, reviews, letters, case reports and
book chapters were excluded. The remaining studies were
potentially eligible and their full text was retrieved. To identify
additional relevant studies, the reference lists of the included
studies were checked manually. Studies were included if they
met the following criteria: (1) sufficient data was reported to
construct 2 × 2 tables (true positive, false positive, true nega-
tive and false negative); (2) the imaging technique was com-
pared with a reference standard (e.g. surgery, histology, fol-
low-up). Exclusion criteria were: (1) evaluation of imaging
techniques other than the aforementioned (e.g. PET-CT,
EUS-FNA, EUS-elastography); (2) imaging techniques used
for treatment of patients with CP (e.g. therapeutic ERCP,
EUS-guided pseudocyst drainage); (3) in vitro studies; (4)
studies that included less than five patients with CP; (5) stud-
ies where no separate analysis were done for patients with CP;
and (6) full-text articles that were not available or retrievable. MEDLINE
N=4243
EMBASE
N=6376
CINAHL
N=300
COCHRANE
N=192
Combined
N=8123
Duplicates
N=2988
Eligible
studies
N=277
Not eligible based
on title and abstract
N=7846
Included
studies
N=43
Excluded on full-text N=234
Reason for exclusion:
- Article not available (n=5)
- Different patient group (n=37)
- In vitro (n=1)
- Insufficient data (n=92)
- Lack of reference standard (n=15)
- Less than 5 patients (n=4)
- Only data on sensitivity (n=33)
- Other disease (n=14)
- Other imaging modality (n=11)
- Other type of article (n=22)
Fig. 1 Flow chart Not eligible based
on title and abstract
N=7846 Introduction Two reviewers in-
dependently (YI and MAK) assessed the QUADAS-2
and the GRADE scoring system and all disagreements
were resolved by reaching consensus. Data analysis calculated. Sensitivity and specificity estimates, the positive
predictive value and negative predictive values, and the accu-
racy were calculated from the reconstructed contingency ta-
bles. We used the I2 test with 95% confidence interval (95%
CI) to quantify heterogeneity [11]. Mean logit sensitivity and
specificity were acquired, and the anti-logit transformation
was then obtained to calculate summary estimates of Overall diagnostic accuracy For each included study we constructed a 2 × 2 contingency
table for each imaging modality. If diagnostic accuracy was
compared between different observers, mean values were Table 1
Study characteristics of included studies
Study
Year
Country
P/R
OE
Modality
Reference standard for CP diagnosis
Adamek et al
2000
Germany
P
No
MRCP/ERCP
Histology (NA), FU (NA)
Albashir et al
2010
USA
R
Yes
EUS
Histology (all)
Alcaraz et al
2000
Spain
P
Yes
MRCP
Surgery (4), ERCP (70), PTC (7)
Balci et al
2006
USA and Germany
R
No
MRCP
ePFT (all)
Bolog et al
2004
Romania
R
No
MRCP
Surgery (NA), ERCP (NA), FU (NA)
Brand et al
2000
Germany
P
No
EUS
Histology (all)
Buscail et al
1995
France
P
No
US/CT/ERCP
/EUS
Histology (7), morphological
changes (i.e. Data extraction and critical appraisal Data was extracted systematically from the included studies
by using a structured study record form. The following study
design and patient characteristics were extracted: name of the
first author, country of origin, year of publication, name of
journal, study design, total number of patients included, num-
ber of included patients with CP, median or mean age, the
proportion of male patients, and the patient inclusion criteria. 3822 Eur Radiol (2017) 27:3820–3844 P prospective, R retrospective, OE observer experience reported, PTC percutaneous transhepatic cholangiogram, ePFT endoscopic pancreatic function
test, FU follow-up, NA not available Data analysis In cases where a negative covariance between the
logit sensitivity and logit specificity was obtained, summary
receiver operating characteristic curve (sROC) were generated
for each separate imaging modality. We used the z test to sensitivity and specificity with 95% CIs. Forest plots were
made to visualize the sensitivity and specificity with the
95% CIs. Data analysis calcifications) and
exocrine insufficiency (42) + FU (all)
Catalano et al
1998
USA
P
No
EUS
ERCP + ePFT (all)
Chong et al
2007
USA
R
Yes
EUS
Surgery (all)
Conwell et al
2007
USA
R
Yes
EUS
ePFT (all)
Dramaix et al
1980
France
P
No
US/CT
Surgery (NA), ERCP (NA)
Fusari et al
2010
Italy
P
Yes
CT/MRCP
Biopsy (33), histology (7)
Gebel et al
1985
Germany
P
No
US/ERP
Obduction (NA), Surgery (NA), FU (NA)
Giovannini et al
1994
France
P
No
EUS
ERCP (all)
Glasbrenner et al
2000
Germany
P
Yes
EUS/ERCP
Surgery (all)
Gmelin et al
1981
Germany
P
No
US/CT/ERCP
Surgery (NA)+FU (NA)
Hellerhoff et al
2002
Germany
P
Yes
MRCP/sMRCP
ERCP (35), surgery (4), FU (56)
Imdahl et al
1999
Germany
P
Yes
CT
Histology (42), FU (6)
Kremer et al
1977
Germany
R
No
US
Clinical diagnosis (338), ERCP, surgery,
ePFT, angiography (NA)
Lammer et al
1980
Germany
R
No
ERCP/CT
Surgery (31), angiography (16),
clinical diagnosis (60)
Lawson et al
1978
USA
R
Yes
ERCP/US
Surgery (25), FU (50)
Lees et al
1979
UK
P
No
US
Surgery (36), ERCP (46)
Lin et al
1989
Taiwan
R
No
US/EUS
Histology (26), CT (4), surgery+ERCP (3)
Nattermann et al
1993
Germany
P
No
EUS
ERCP (94), FU (20)
Pamos et al
1998
Spain
P
Yes
MRCP
ERCP (all)
Parsi et al
2008
USA
R
Yes
ERCP
FU (all)
Pistolesi et al
1981
Italy
P
No
CT
Surgery (all)
Pungpapong et al
2007
USA
P
Yes
EUS
Clinical history, lab data, ERCP/CT/MRI
and/or surgical pathology (all)
Pungpapong et al
2007
USA
P
Yes
MRCP/EUS
ERCP (48), surgery (9), FU (57)
Rudowicz-Pietruszewska et al
2002
Poland
P
No
MRCP
ERCP (all)
Sai et al
2008
Japan
P
Yes
sMRCP
ERCP (all)
Savarino et al
1980
Italy
R
No
CT
Surgery (NA), calcifications (NA), clinical and lab data (NA)
Scarabino et al
1989
Italy
R
No
ERCP, US, CT
Combination of CT, US and ERCP (all)
Schlaudraff et al
2008
USA and Germany
P
Yes
MRCP/sMRCP
Clinical history, laboratory, radiology (≥2 methods) (all)
Stevens et al
2009
USA
P
Yes
EUS
ePFT (all)
Sverko et al
2011
Croatia
R
No
MRCP
Histology (all)
Swobodnik et al
1983
Germany
P
No
US/CT/ERCP
FU (59), surgery (22)
Tox et al
2007
Germany
R
Yes
EUS
Surgery (79), FU (92)
Trikudanathan et al
2016
USA
R
YES
EUS
Histology (all)
Triller et al
1975
Switzerland
P
No
ERCP
Surgery (14), autopsy (1), FU (9)
Wiersema et al
1993
USA
P
No
EUS/ERCP
FU (51), ePFT (16)
Zhang et al
2003
USA
R
No
MRCP
US (12), CT (11), ERCP (6)
Zuccaro et al
2009
USA
R
No
MRCP/sMRCP
ePFT (all)
P prospective, R retrospective, OE observer experience reported, PTC percutaneous transhepatic cholangiogram, ePFT endoscopic pancreatic function
t t FU f ll
NA
t
il bl Table 1
Study characteristics of included studies 3823 Eur Radiol (2017) 27:3820–3844 model [12]. Study and patient characteristics Study characteristics, including the reference standard for the
diagnosis of CP for each included study, are listed in Table 1. The 43 included studies were published between 1975 and
2016; 26 studies were prospective and 23 studies were pub-
lished after the year 2000. A total of 3460 patients were eval-
uated, of which 1242 patients were diagnosed with CP
[14–56]. The age of the patients ranged from 36 to 65 years,
with a median of 50% male. Criteria for selection of patients
were those with suspected pancreatic disease or patients with
suspected CP. Patient characteristics are depicted in Table 2. Heterogeneity exploration Heterogeneity exploration The following factors were incorporated in the bivariate mod-
el and we evaluated the effect on the sensitivity and specific-
ity, and cause of heterogeneity for all imaging modalities ac-
cording to the QUADAS-2 tool: clear description of criteria
for bias (low bias versus high bias or unclear) for (a) patient
selection, (b) criteria for the index test used, (c) sufficient
description and verification with the reference standard, and
(d) the flow and timing. Data analysis 2 Summary of study quality
(QUADAS-2) 0%
20%
40%
60%
80%
100%
PATIENT SELECTION
INDEX TEST
REFERENCE STANDARD
FLOW AND TIMING
Proportion of studies with low, high or unclear
RISK of BIAS
0%
20%
40%
60%
80%
100%
Proportion of studies with low, high, or unclear
CONCERNS regarding APPLICABILITY
Low
High
Unclear
Fig. 2 Summary of study quality
(QUADAS-2) Fig. 2 Summary of study quality
(QUADAS-2) 0%
20%
40%
60%
80%
100%
Proportion of studies with low, high or unclear
RISK of BIAS
0%
20%
40%
60%
80%
100%
Proportion of studies with low, high, or unclear
CONCERNS regarding APPLICABILITY evaluate differences in sensitivity and specificity between the
five imaging modalities. A p value of less than 0.05 indicated
a statistically significant difference. Study selection The initial search resulted in 11,111 hits, of which 2988 du-
plicates were removed, resulting in a total of 8123 titles and
abstracts that were screened for eligibility. The full text of 277
articles was retrieved; 43 of these articles fulfilled the inclu-
sion criteria. See Appendix Table 6 for the excluded articles. Figure 1 shows the flow chart of the search. Data analysis Summary estimates of sensitivity and specificity,
including 95% CI, were obtained by using a random-effects Table 2
Patient characteristics of included studies Table 2
Patient characteristics of included studies
Study
Nr pts
Age
Male (%)
Nr pts CP
Patient selection
Adamek et al
124
55
61%
57
Suspected pancreatic mass (clinical presentation, lab, US)
Albashir et al
23
43*
57%
19
Suspected chronic pancreatitis (clinical presentation)
Alcaraz et al
81
65**
31%
8
Suspected pancreatobiliary disease (clinical presentation, US)
Balci et al
30
48*
17%
11
Suspected early CP (clinical presentation)
Bolog et al
103
57*
43%
15
Suspected pancreatobiliary disease (US/CT or clinical presentatio
Brand et al
115
61*
59%
24
Suspected focal pancreatic lesion (US/CT/ERCP or lab/tumour
markers)
Buscail et al
62
50*
79%
44
Suspected chronic pancreatitis (clinical presentation,
lab, imaging)
Catalano et al
80
51*
40%
38
Non-alcoholic recurrent acute pancreatitis (3–11 episodes)
Chong et al
71
45*
46%
64
Suspected chronic pancreatitis (clinical presentation)
Conwell et al
56
44*
45%
38
Suspected chronic pancreatitis (clinical presentation)
Dramaix et al
50
52*
66%
18
Suspected pancreatic disease (clinical presentation)
Fusari et al
40
62*
55%
8
Suspected pancreatic mass (clinical presentation and US)
Gebel et al
US: 56, ERP: 45
NA
NA
US: 22, ERP: 16
Suspected pancreatic disease (clinical presentation)
Giovannini et al
26
NA
NA
17
Suspected pancreatobiliary disease (clinical
presentation, imaging/lab)
Glasbrenner et al
85
NA
NA
41
Suspected pancreatic mass (clinical presentation, US/CT)
Gmelin et al
41
54*
68%
19
Suspected pancreatic disease (clinical presentation)
Hellerhoff et al
95
NA
NA
26
Suspected pancreatic disease (clinical presentation)
Imdahl et al
48
58*
60%
12
Suspected pancreatic disease (clinical presentation)
Kremer et al
446
NA
NA
61
Suspected pancreatic disease (clinical presentation)
Lammer et al
107
NA
NA
39
Suspected pancreatic disease (clinical presentation)
Lawson et al
75
NA
NA
26
Suspected pancreatic disease (clinical presentation)
Lees et al
98
NA
NA
20
Suspected pancreatic disease (clinical presentation)
Lin et al
33
47*
58%
7
Suspected pancreatic disease (clinical presentation)
Nattermann et al
114
53*
67%
51
Suspected pancreatic disease (clinical presentation)
Pamos et al
41
64*
59%
5
Suspected pancreatobiliary disease (clinical presentation)
Parsi et al
35
46**
46%
24
Suspected chronic pancreatitis (clinical presentation)
Pistolesi et al
100
NA
NA
31
Suspected pancreatic disease (clinical presentation)
Pungpapong et al
79
50**
35%
38
Suspected chronic pancreatitis (clinical presentation)
Pungpapong et al
99
55**
41%
40
Suspected chronic pancreatitis (clinical presentation)
Rudowicz-Pietruszewska et al
88
52*
64%
9
Suspected pancreatobiliary disease (clinical presentation,
lab, US/CT)
Sai et al
28
36*
NA
16
Mild chronic pancreatitis (ERCP)
Savarino et al
108
47**
67%
59
Suspected pancreatic disease (clinical presentation)
Scarabino et al
63
44**
63%
12
Suspected of biliopancreatic disease (clinical presentation)
Schlaudraff et al
62
NA
NA
9
Suspected chronic pancreatitis (clinical presentation)
Stevens et al
100
NA
38%
41
Suspected chronic pancreatitis (clinical presentation)
Sverko et al
29
44**
52%
14
Suspected pancreatic disease (clinical presentation)
Swobodnik et al
81
49*
52%
27
Suspected pancreatic disease (clinical presentation)
Tox et al
171
61*
NA
65
Suspected pancreatic disease (clinical presentation)
Trikudanathan et al
68
39*
18%
56
Total pancreatectomy for non-calcific chronic pancreatitis
Triller et al
24
52*
83%
11
Suspected pancreatobiliary disease (clinical presentation)
Wiersema et al
67
45*
20%
30
Suspected pancreatobiliary disease (clinical presentation)
Zhang et al
44
50*
30%
24
Suspected early or mild chronic pancreatitis (clinical
presentation, US/CT/ERCP)
Zuccaro et al
69
43*
35%
28
Suspected chronic pancreatitis (clinical presentation)
NA not available 3824 Eur Radiol (2017) 27:3820–3844 0%
20%
40%
60%
80%
100%
PATIENT SELECTION
INDEX TEST
REFERENCE STANDARD
FLOW AND TIMING
Proportion of studies with low, high or unclear
RISK of BIAS
0%
20%
40%
60%
80%
100%
Proportion of studies with low, high, or unclear
CONCERNS regarding APPLICABILITY
Low
High
Unclear
Fig. Fig. 3 Forest plot for sensitivity and specificity Head to head comparison A head to head comparison was performed in studies that
compared the diagnostic accuracy of two or more imaging
modalities. Heterogeneity was quantified by I2 test, with
95% CI. The random-effects (I2 > 25%) and fixed effects (I2
≤25%) models were used to obtain summary estimates of
sensitivity and specificity, and compared with one another
by a paired z test. The risk of bias, assessed by QUADAS-2, was low in 28%
of the studies and high in 19% of the studies. The concerns
about applicability were low in 30% of the studies and high in
40% of the studies. The QUADAS-2 characteristics for each
domain are depicted in Fig. 2 and outlined for each study in
Appendix Table 7. The quality of evidence for all five imaging
modalities according to the GRADE scoring system was very For data analysis, Review Manager (RevMan, version 5.3. Copenhagen: The Cochrane Collaboration, 2014) and SAS
(version 9.3; SAS Institute, Cary, NC) were used. We adhered
to the Preferred Reporting Items for Systematic Reviews and
Meta-Analyses (PRISMA) guidelines [13]. Table 3 Estimated overall
sensitivity, specificity and
heterogeneity according to
imaging modality
Modality
N studies
N patients
Sensitivity (95% CI)
Specificity (95% CI)
Heterogeneity (I2)
EUS
16
1249
81% (70–89%)
90% (82–95%)
82%/73%
ERCP
11
742
82% (76–87%)
94% (87–98%)
39%/67%
MRCP
14
933
78% (69–85%)
96% (90–98%)
59%/65%
CT
10
700
75% (66–83%)
91% (81–96%)
50%/71%
US
10
1005
67% (53–78%)
98% (89–100%)
40%/93%
Random effects model Modality
N studies
N patients
Sensitivity (95% CI)
Specificity (95% CI)
Heterogeneity (I2)
EUS
16
1249
81% (70–89%)
90% (82–95%)
82%/73%
ERCP
11
742
82% (76–87%)
94% (87–98%)
39%/67%
MRCP
14
933
78% (69–85%)
96% (90–98%)
59%/65%
CT
10
700
75% (66–83%)
91% (81–96%)
50%/71%
US
10
1005
67% (53–78%)
98% (89–100%)
40%/93%
Random effects model 3825 Eur Radiol (2017) 27:3820–3844 Forest plot: EUS
Forest plot: ERCP
Forest plot: MRCP
Forest plot CT
Forest plot: US
for sensitivity and specificity 3826 Eur Radiol (2017) 27:3820–3844 low. The GRADE scores for each imaging modality and char-
acteristics for each study are outlined in Appendix Tables 8
and 9. Heterogeneity exploration The bivariate model for heterogeneity exploration showed that
the factor ‘flow and timing’ was significantly associated with
a higher sensitivity of US (p = 0.01). ‘Description and verifi-
cation with the reference standard’ was significantly associat-
ed with a higher specificity for MRCP (p = 0.0002). Head to head comparison The head to head comparison of US versus ERCP compar-
ison yields a sensitivity of 57% (49–65%) versus 78% (71–
85%) (p < 0.001); and a specificity of 94% (74–99%) versus EUS was the most frequently evaluated imaging modality;
16 studies including 1249 patients [15, 19–23, 27, 28, 36, 37,
41, 42, 48, 51, 53, 56]. ERCP was studied in 11 studies in-
cluding 742 patients [14, 20, 26, 28, 29, 33, 34, 39, 46, 50,
52]; MRCP, including secretin-enhanced MRCP, was evaluat-
ed in 14 studies including 933 patients [14, 16–18, 25, 30, 38,
42–44, 47, 49, 54, 55]; CT in 10 studies including 700 patients
[20, 24, 25, 29, 31, 33, 40, 45, 46, 50] and abdominal US in 10
studies which included 1005 patients [20, 24, 26, 29, 32,
34–36, 46, 50]. The imaging characteristics for each study
and modality in an individual study are listed in Appendix
Table 11. Three of the 43 articles reported about complications
of the imaging modality used; these were complications relat-
ed to ERCP (being post-ERCP pancreatitis) with a mean com-
plication rate of 4% [14, 20, 28]. EUS
ERCP EUS
ERCP
CT
Fig. 4 Receiver operator curves (ROC) Overall diagnostic accuracy Analyses for summary estimates of sensitivity and specificity
were done for EUS, ERCP, MRI, CT and US (Table 3). Figures 3 and 4 show sensitivity and specificity of individual
studies in forest plots and in receiver operator curves (ROC),
respectively. A negative covariance between the logit sensi-
tivity and logit specificity was not obtained; therefore, no
sROC for MRI and US could be drawn. The summary esti-
mate of sensitivity for EUS, ERCP, MRCP, CT and US was
81%, 82%, 78%, 75% and 67%, respectively. The summary
estimate of specificity for EUS, ERCP, MRCP, CT and US
was 90%, 94%, 96%, 91% and 98%, respectively. Sensitivity of ERCP was significant higher than sensitivity
of US (p = 0.018). Other pairwise comparisons of sensitivity
between imaging modalities revealed no significant diffe-
rence. Specificity did not differ significantly among all
modalities (Table 3). Sensitivity and specificity values for
each study are listed in Appendix Table 10. CT CT CT
Fig. 4 Receiver operator curves (ROC) Head to head comparison Six head to head comparisons were performed (Table 4). The
specificity of ERCP and EUS, and the sensitivity of ERCP,
EUS and CT in the summary estimates of the head to head
studies were significantly higher as compared with US. Fig. 4 Receiver operator curves (ROC) Eur Radiol (2017) 27:3820–3844 3827 98% (89–100%) (p = 0.003), respectively [20, 26, 29, 34, 46,
50]. The comparison between US and CT yields a sensitivity
of 58% (49–66%) and 77% (68–83%) (p = 0.002), respective-
ly [20, 24, 29, 46, 50]. And finally, the comparison of EUS
versus US comparison yields a sensitivity of 90% (82–98%)
versus 63% (49–76%) (p = 0.001); and a specificity of 100%
versus 91% (82–99%) (p = 0.04), respectively [20, 36]. There
were no significant differences in the sensitivity and specific-
ity estimates between ERCP and EUS [20, 28, 53], MRCP
and sMRCP [30, 47, 55] or ERCP and CT [20, 29, 33, 46, 50]. The heterogeneity (I2) between US and ERCP (>25%) was
higher (>25%) than in the other comparisons (I2 ≤25%). and 78%, respectively, in the present meta-analyses. The
European Society of Radiology (ESR) is developing the
ESR iGuide, a clinical decision support system for European
imaging referral guidelines, covering various clinical scenari-
os, indications and recommendations (www.esriguide.org)
[61–63]. The results from the present systematic review may
be useful to incorporate in that system. We excluded three studies where sensitivity and specificity
data were provided, but it was not possible to extract sufficient
data to produce 2 × 2 tables and calculate the diagnostic ac-
curacy values, because only the sensitivity and specificity es-
timates were given [64–66]. In the study by Wang et al., esti-
mates of sensitivity and specificity for EUS, ERCP and US
were in line with the present results; the sensitivity of MR
imaging and CT, however, were much lower (66% and
61%) [66]. The studies by Clave et al. and Orti et al. showed
a lower sensitivity of ERCP (62% and 70%, respectively)
compared to present results (82%) [64, 65]. Discussion EUS, ERCP, MRI and CTall have comparable high diagnostic
accuracy in the initial diagnosis of chronic pancreatitis. EUS
and ERCP are outperformers and US has the lowest accuracy. The choice of imaging modality can therefore be made on the
basis of invasiveness, local availability, experience and costs. Several recent guidelines [57–59] advocate the use of EUS,
MRCP or CT for the diagnosis of CP, although summary
estimates of their accuracy, thus far, were lacking. There is
one guideline from Germany on CP that has reported sensi-
tivity and specificity regarding EUS, ERCP, MRCP and US,
although not for CT [60]. In this guideline 14 studies were
selected, reporting ranges rather than pooling the data on sen-
sitivity and specificity estimates. This method resulted in re-
sults slightly different from those in the present meta-analyses. For example the guideline reports a sensitivity of 70–80% for
ERCP and 88% for MRI versus summary estimates of 82% EUS, ERCP, MRI and CTall have comparable high diagnostic
accuracy in the initial diagnosis of chronic pancreatitis. EUS
and ERCP are outperformers and US has the lowest accuracy. The choice of imaging modality can therefore be made on the
basis of invasiveness, local availability, experience and costs. The risk of missing important studies was minimized by
performing a search in four major databases by two reviewers
independently, without setting any restrictions for language
and publication date. However, this systematic review has
some limitations. The heterogeneity of the pooled studies
was moderate to high in all analyses (between 39% and
93%). However, in the head to head comparison analyses,
the heterogeneity was low in most comparisons (<25%). Furthermore, the heterogeneity of the reference standards used
in the studies could have influenced individual study results. Surgery, histology and long-term follow-up of patients are
reliable methods. Some reference standards, such as the use
of endoscopic pancreatic function test (ePFT) for establishing
the diagnosis of CP, could have resulted in under- or Several recent guidelines [57–59] advocate the use of EUS,
MRCP or CT for the diagnosis of CP, although summary
estimates of their accuracy, thus far, were lacking. There is
one guideline from Germany on CP that has reported sensi-
tivity and specificity regarding EUS, ERCP, MRCP and US,
although not for CT [60]. In this guideline 14 studies were
selected, reporting ranges rather than pooling the data on sen-
sitivity and specificity estimates. Table 4 Head to head
comparison Sensitivity: US vs ERCP (p < 0.001), US vs CT (p = 0.002), EUS vs US (p = 0.001) Discussion This method resulted in re-
sults slightly different from those in the present meta-analyses. For example the guideline reports a sensitivity of 70–80% for
ERCP and 88% for MRI versus summary estimates of 82% Table 4 Head to head
comparison Comparison
N studies
N patients
Modality
Sensitivity (95% CI)
Specificity (95% CI)
US vs ERCPa
6
423
US
57% (49–65%)
94% (74–99%)
ERCP
78% (71–85%)
98% (89–100%)
US vs CTb
5
297
US
58% (49–66%)
77% (71–83%)
CT
77% (68–83%)
82% (74–88%)
CT vs ERCPb
5
354
CT
75% (67–82%)
86% (81–90%)
ERCP
84% (77–89%)
90% (85–93%)
EUS vs ERCPb
3
214
EUS
88% (80–93%)
85% (76–91%)
ERCP
86% (78–91%)
92% (85–96%)
MRCP vs sMRCPb
3
226
MRCP
62% (49–73%)
94% (89–97%)
sMRCP
68% (56–79%)
91% (85–94%)
EUS vs USb
2
95
EUS
90% (82–98%)
100%
US
63% (49–76%)
91% (82–99%)
Sensitivity: US vs ERCP (p < 0.001), US vs CT (p = 0.002), EUS vs US (p = 0.001)
Specificity: US vs ERCP (p = 0.003), EUS vs US (p = 0.04)
a Random effects model
b Fixed effects model 3828 Eur Radiol (2017) 27:3820–3844 overestimation of the sensitivity and specificity. In addition,
the diagnosis of CP and the criteria used are different in dif-
ferent stages of the disease (e.g. absence of calcifications in
the early phase of the disease). Another limitation was that our
analyses included imaging studies and imaging protocols per-
formed over the last 40 years in different centres with inherent
variations in techniques and equipment. Especially in the last
decade the quality of some imaging modalities (e.g. MRCP
and CT) has improved considerably. Also there were concerns
about the quality of the available evidence, as assessed by
QUADAS-2 and the GRADE scoring system. (e.g. PD dilatation and strictures) and slight changes of the
pancreatic parenchyma and side branches, which can be at-
tributed to early signs CP (i.e. atrophy, side branch ectasia)
compared to CT [74]. Early diagnosis can also lead to a timely
start of treatment, which has been associated with improved
long-term outcome [75]. Nevertheless, for very early CP this
association needs to be established in further research, such as
the ESCAPE trial, evaluating the effect of early intervention in
patients with CP [76]. Discussion As diagnostic sensitivity of CT and
MRCP is not significantly lower than that of ERCP and
EUS, and specificity is comparable, non-invasive modalities
except for US are a likely first choice in patients with
suspected pancreatic disease including chronic pancreatitis. The highest scores for accuracy in the diagnosis of CP were
found for EUS and ERCP, but these are invasive techniques. ERCP has a relatively high risk of complications, such as post-
ERCP pancreatitis (1.6–15.7%, mean complication rate of
4%) and is nowadays only used for therapeutic purposes
(e.g. stenting of pancreatic duct) [67–69]. To date, diagnostic
ERCP is largely replaced by EUS and the cross-sectional im-
aging modalities CT and MRCP. Acknowledgements
The scientific guarantor of this publication is
M.A. Boermeester. The authors of this manuscript declare relationships
with the following companies: Baxter, Acelity/KCI, Ipsen, Mylan,
Johnson & Johnson, Bard. The authors of this manuscript declare no
relationships with any companies whose products or services may be
related to the subject matter of the article. The authors state that this work
has not received any funding. Two of the authors have significant statis-
tical expertise (SB, MAB). Institutional review board approval was not
required because of the nature of the study (a systematic review). Acknowledgements
The scientific guarantor of this publication is
M.A. Boermeester. The authors of this manuscript declare relationships
with the following companies: Baxter, Acelity/KCI, Ipsen, Mylan,
Johnson & Johnson, Bard. The authors of this manuscript declare no
relationships with any companies whose products or services may be
related to the subject matter of the article. The authors state that this work
has not received any funding. Two of the authors have significant statis-
tical expertise (SB, MAB). Institutional review board approval was not
required because of the nature of the study (a systematic review). It has been suggested that CT is better in detecting paren-
chymal calcifications and intraductal calcifications compared
to MRCP [70–73]. On the other hand, MRCP is more often
able to detect significant abnormalities of the pancreatic duct MeSH Medical Subject Headings APPENDIX Table 5 Search terms Table 5 Search terms MeSH terms
All Fields
Chronic
pancreatitis
Pancreatitis, chronic [MeSH]
Chronic pancreatitis [All Fields]
AND
EUS
Endosonography [MeSH]
EUS [All Fields]
OR
ERCP
Cholangiopancreatography, Endoscopic
Retrograde [MeSH]
Endoscopic Retrograde Cholangiopancreatograp*
[All Fields] OR ERCP [All Fields]
OR
MRCP
Magnetic Resonance Imaging [MeSH]
OR
Cholangiopancreatography, Magnetic
Resonance [MeSH]
Magnetic resonance imaging [All Fields]
OR MRI [All Fields] OR MRCP [All Fields]
OR Magnetic Resonance Cholangio* [All
Fields]
OR
sMRCP
Magnetic Resonance Imaging [All Fields]
AND secretin [All Fields] OR sMRI [All Fields]
OR
CT
Tomography, X-Ray Computed [MeSH]
(Tomography [All Fields] AND x-ray
[All Fields] AND computed [All Fields])
OR Computed Tomography [All Fields])
OR CT scan* [All Fields]
OR
US
Ultrasonography [MeSH]
Ultrasonogra* [All Fields] OR ultrasound [All
Fields]
MeSH Medical Subject Headings EUS [All Fields] 3829 Eur Radiol (2017) 27:3820–3844 Author
Year
Journal
Borsukov et al
2001
Ross Gastroenterol Zh
Diad'kin et al
2013
Vestnik rentgenologii i radiologii
Dotsenko et al
1985
Vrach Delo
Rosch et al
1989
Z Gastroenterologie
Suzdalev et al
1992
Likars'ka sprava
Agarwal et al
2008
GIE
Brailski et al
1989
Vutr Boles
Brailski et al
1984
Vutr Boles
Brimiene et al
2011
Medicina
Carlucci et al
1989
HPB Surgery
Chowdhury et al
2005
Pancreas
Cotton et al
1980
Radiology
DelMaschio et al
1991
Radiology
Erturk et al
2006
Am J Gastroenterol
Frick et al
1982
Gastrointest Rad
Gheonea et al
2013
BMC Gastroenterology
Goodale et al
1981
Ann Surg
Hanninen et al
2002
Radiology
Hatano et al
1998
Nippon rinsho J
Hocke et al
2008
Dtsch Med Wochenschr
Hocke et al
2006
WJG
Hocke et al
2012
Z Gastroenterologie
Huang et al
2011
J Dig dis
Imbriaco et al
2006
Radiol Med
Kawai et al
2012
Eur J Rad
Kim et al
2007
J MRI
Kursawa et al
1991
Radiol Diagn
Lu et al
2013
Acad J Sec Mil Med University
Lutz et al
1975
Klin Wschr
Morris-Stiff et al
2009
J Pancreas
Papp et al
1978
Wiener klin Wchnschrft
Pomerri et al
1991
Radiologia Med
Rosch et al
2000
Am J Gastroenterol
Sandrasegaran et al
2013
AJR
Sendler et al
2000
World J Surg
Sugumar et al
2011
Gut
Testoni et al
1981
Acta Endoscopica
Tiushin et al
2003
Voprosy onkologii
Varadarajulu et al
2007
GIE
Viceconte et al
1980
Ann ital chir
Yamada et al
2010
Abdom Imaging
Zhu et al
2013
PLOS one
Bhutani et al
2009
Pancreas
Akisik et al
2013
AJR
Alempijević et al
2005
Vojnosanit Pregl
Alpern et al
1985
Radiology
Ardelean et al
2014
Med Ultrason
Ardengh et al
2011
GIE
Ascunce et al
2010
Surg End
Baert et al
1977
Radiologe
Balci et al
2010
J MRI
Beliao et al
2012
Eur J Rad
Bender et al
1999
Invest Rad
Bhatt et al
2005
Indian J Rad Imag Ass
Bonanno et al
1994
Giorn Ital End Dig
Bruhlmann et al
1976
RoFo
Caletti et al
1982
British j Surgery
Cao
1989
Zhonghua yi xue za zhi
Cappeliez et al
2000
Radiology
Chang et al
2010
GIE Author
Year
Journal
Reason for exclusion
Borsukov et al
2001
Ross Gastroenterol Zh
Article not available
Diad'kin et al
2013
Vestnik rentgenologii i radiologii
Article not available
Dotsenko et al
1985
Vrach Delo
Article not available
Rosch et al
1989
Z Gastroenterologie
Article not available
Suzdalev et al
1992
Likars'ka sprava
Article not available
Agarwal et al
2008
GIE
Exclusive patient group
Brailski et al
1989
Vutr Boles
Exclusive patient group
Brailski et al
1984
Vutr Boles
Exclusive patient group
Brimiene et al
2011
Medicina
Exclusive patient group
Carlucci et al
1989
HPB Surgery
Exclusive patient group
Chowdhury et al
2005
Pancreas
Exclusive patient group
Cotton et al
1980
Radiology
Exclusive patient group
DelMaschio et al
1991
Radiology
Exclusive patient group
Erturk et al
2006
Am J Gastroenterol
Exclusive patient group
Frick et al
1982
Gastrointest Rad
Exclusive patient group
Gheonea et al
2013
BMC Gastroenterology
Exclusive patient group
Goodale et al
1981
Ann Surg
Exclusive patient group
Hanninen et al
2002
Radiology
Exclusive patient group
Hatano et al
1998
Nippon rinsho J
Exclusive patient group
Hocke et al
2008
Dtsch Med Wochenschr
Exclusive patient group
Hocke et al
2006
WJG
Exclusive patient group
Hocke et al
2012
Z Gastroenterologie
Exclusive patient group
Huang et al
2011
J Dig dis
Exclusive patient group
Imbriaco et al
2006
Radiol Med
Exclusive patient group
Kawai et al
2012
Eur J Rad
Exclusive patient group
Kim et al
2007
J MRI
Exclusive patient group
Kursawa et al
1991
Radiol Diagn
Exclusive patient group
Lu et al
2013
Acad J Sec Mil Med University
Exclusive patient group
Lutz et al
1975
Klin Wschr
Exclusive patient group
Morris-Stiff et al
2009
J Pancreas
Exclusive patient group
Papp et al
1978
Wiener klin Wchnschrft
Exclusive patient group
Pomerri et al
1991
Radiologia Med
Exclusive patient group
Rosch et al
2000
Am J Gastroenterol
Exclusive patient group
Sandrasegaran et al
2013
AJR
Exclusive patient group
Sendler et al
2000
World J Surg
Exclusive patient group
Sugumar et al
2011
Gut
Exclusive patient group
Testoni et al
1981
Acta Endoscopica
Exclusive patient group
Tiushin et al
2003
Voprosy onkologii
Exclusive patient group
Varadarajulu et al
2007
GIE
Exclusive patient group
Viceconte et al
1980
Ann ital chir
Exclusive patient group
Yamada et al
2010
Abdom Imaging
Exclusive patient group
Zhu et al
2013
PLOS one
Exclusive patient group
Bhutani et al
2009
Pancreas
In vitro
Akisik et al
2013
AJR
No diagnostic values for CP
Alempijević et al
2005
Vojnosanit Pregl
No diagnostic values for CP
Alpern et al
1985
Radiology
No diagnostic values for CP
Ardelean et al
2014
Med Ultrason
No diagnostic values for CP
Ardengh et al
2011
GIE
No diagnostic values for CP
Ascunce et al
2010
Surg End
No diagnostic values for CP
Baert et al
1977
Radiologe
No diagnostic values for CP
Balci et al
2010
J MRI
No diagnostic values for CP
Beliao et al
2012
Eur J Rad
No diagnostic values for CP
Bender et al
1999
Invest Rad
No diagnostic values for CP
Bhatt et al
2005
Indian J Rad Imag Ass
No diagnostic values for CP
Bonanno et al
1994
Giorn Ital End Dig
No diagnostic values for CP
Bruhlmann et al
1976
RoFo
No diagnostic values for CP
Caletti et al
1982
British j Surgery
No diagnostic values for CP
Cao
1989
Zhonghua yi xue za zhi
No diagnostic values for CP
Cappeliez et al
2000
Radiology
No diagnostic values for CP
Chang et al
2010
GIE
No diagnostic values for CP Table 6 Excluded articles based
on full text Eur Radiol (2017) 27:3820–3844 3830 Author
Year
Journal
Cohen et al
2014
Dig Dis Sci
Concia et al
2014
Invest Rad
Dale et al
1979
Electromedica
Das et al
2008
GIE
Delbeke et al
1999
J Nucl Med
Dite et al
1982
Vnitrni Lekarstvi
Dronamraju et al
2016
Ann Gastroenterol
D’Souza et al
2015
Dig Dis Sci
Eitner et al
1979
Dtsch Zeitschr Verdauungs- und
Stoffwechselkrankheiten
Eloubeidi et al
2013
Pancreas
Ergul et al
2014
Rev Esp Med Nucl Im Mol
Ferrucci et al
1979
Radiology
Foley et al
1980
Gastrointest Rad
Fontana et al
1976
Gut
Foster et al
1984
BMJ
Gardner et al
2014
Pancreas
Gincul et al
2014
Endoscopy
Gowland et al
1981
Lancet
Grant et al
1981
J Am Osteopathic Ass
Harada et al
1977
Gastroenterologica Jap
He et al
2014
Pancreas
Hoki et al
2009
J Gastroenterol
Hollerbach et al
1994
Med Klinik
Horii et al
1982
Jap J Gastroenterol
Johnson et al
1999
Radiology
Jones et al
1988
Clin Radiol
Kamisawa et al
2007
J Gastroenterol
Kersting et al
2009
Gastroenterology
Kitano et al
2004
Gut
Laghi et al
1998
Chirurgia
Leblanc et al
2014
Pancreas
Leblanc et al
2014
Pancreas
Li et al
2001
Zhongguo yi xue ke xue
Loginov et al
1976
Sovetskaya Meditsina
Lopez et al
2002
Radiology
Manfredi
2000
Radiology
Modder et al
1979
RoFo
Montori et al
1979
Min Diet Gastroent
Napoleon et al
2010
Endoscopy
Novis et al
1976
S Afr Med J
Ohtsubo et al
2008
Gastroenterolog Endoscopy
Orlikov et al
2007
Ter Arkh
Park et al
2008
The Korean J Gastroenter
Petersein et al
2002
RoFo
Pezzelli et al
2013
Pancreas
Pomerri et al
1987
Radiologia Med
Rickes et al
2002
Scand J Gastroenterol
Rosenberger et al
1979
MMW
Russell et al
1978
Gut
Sahai et al
1998
GIE
Sainani et al
2009
AJG
Sica et al
2002
J MRI
Sica et al
1999
Radiology
Songur et al
2000
Digest Endoscopy
Stevens et al
2010
WJG
Struve et al
1982
Diagnostik & Intensivtherapie
Sun et al
2010
Acad J Sec Mil Med University
Tamura et al
2006
Radiology
Tellez-Avila et al
2014
WJG Author
Year
Journal
Reason for exclusion
Cohen et al
2014
Dig Dis Sci
No diagnostic values for CP
Concia et al
2014
Invest Rad
No diagnostic values for CP
Dale et al
1979
Electromedica
No diagnostic values for CP
Das et al
2008
GIE
No diagnostic values for CP
Delbeke et al
1999
J Nucl Med
No diagnostic values for CP
Dite et al
1982
Vnitrni Lekarstvi
No diagnostic values for CP
Dronamraju et al
2016
Ann Gastroenterol
No diagnostic values for CP
D’Souza et al
2015
Dig Dis Sci
No diagnostic values for CP
Eitner et al
1979
Dtsch Zeitschr Verdauungs- und
Stoffwechselkrankheiten
No diagnostic values for CP
Eloubeidi et al
2013
Pancreas
No diagnostic values for CP
Ergul et al
2014
Rev Esp Med Nucl Im Mol
No diagnostic values for CP
Ferrucci et al
1979
Radiology
No diagnostic values for CP
Foley et al
1980
Gastrointest Rad
No diagnostic values for CP
Fontana et al
1976
Gut
No diagnostic values for CP
Foster et al
1984
BMJ
No diagnostic values for CP
Gardner et al
2014
Pancreas
No diagnostic values for CP
Gincul et al
2014
Endoscopy
No diagnostic values for CP
Gowland et al
1981
Lancet
No diagnostic values for CP
Grant et al
1981
J Am Osteopathic Ass
No diagnostic values for CP
Harada et al
1977
Gastroenterologica Jap
No diagnostic values for CP
He et al
2014
Pancreas
No diagnostic values for CP
Hoki et al
2009
J Gastroenterol
No diagnostic values for CP
Hollerbach et al
1994
Med Klinik
No diagnostic values for CP
Horii et al
1982
Jap J Gastroenterol
No diagnostic values for CP
Johnson et al
1999
Radiology
No diagnostic values for CP
Jones et al
1988
Clin Radiol
No diagnostic values for CP
Kamisawa et al
2007
J Gastroenterol
No diagnostic values for CP
Kersting et al
2009
Gastroenterology
No diagnostic values for CP
Kitano et al
2004
Gut
No diagnostic values for CP
Laghi et al
1998
Chirurgia
No diagnostic values for CP
Leblanc et al
2014
Pancreas
No diagnostic values for CP
Leblanc et al
2014
Pancreas
No diagnostic values for CP
Li et al
2001
Zhongguo yi xue ke xue
No diagnostic values for CP
Loginov et al
1976
Sovetskaya Meditsina
No diagnostic values for CP
Lopez et al
2002
Radiology
No diagnostic values for CP
Manfredi
2000
Radiology
No diagnostic values for CP
Modder et al
1979
RoFo
No diagnostic values for CP
Montori et al
1979
Min Diet Gastroent
No diagnostic values for CP
Napoleon et al
2010
Endoscopy
No diagnostic values for CP
Novis et al
1976
S Afr Med J
No diagnostic values for CP
Ohtsubo et al
2008
Gastroenterolog Endoscopy
No diagnostic values for CP
Orlikov et al
2007
Ter Arkh
No diagnostic values for CP
Park et al
2008
The Korean J Gastroenter
No diagnostic values for CP
Petersein et al
2002
RoFo
No diagnostic values for CP
Pezzelli et al
2013
Pancreas
No diagnostic values for CP
Pomerri et al
1987
Radiologia Med
No diagnostic values for CP
Rickes et al
2002
Scand J Gastroenterol
No diagnostic values for CP
Rosenberger et al
1979
MMW
No diagnostic values for CP
Russell et al
1978
Gut
No diagnostic values for CP
Sahai et al
1998
GIE
No diagnostic values for CP
Sainani et al
2009
AJG
No diagnostic values for CP
Sica et al
2002
J MRI
No diagnostic values for CP
Sica et al
1999
Radiology
No diagnostic values for CP
Songur et al
2000
Digest Endoscopy
No diagnostic values for CP
Stevens et al
2010
WJG
No diagnostic values for CP
Struve et al
1982
Diagnostik & Intensivtherapie
No diagnostic values for CP
Sun et al
2010
Acad J Sec Mil Med University
No diagnostic values for CP
Tamura et al
2006
Radiology
No diagnostic values for CP
Tellez-Avila et al
2014
WJG
No diagnostic values for CP Table 6 (continued) No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP
No diagnostic values for CP No diagnostic values for CP No diagnostic values for CP No diagnostic values for CP No diagnostic values for CP No diagnostic values for CP No diagnostic values for CP No diagnostic values for CP No diagnostic values for CP No diagnostic values for CP No diagnostic values for CP Author
Year
Journal
Tirkes et al
2016
J MRI
Trikudanathan et al
2015
Am J Gastroenterol
Tripathi et al
2002
Indian J Gastroenterol
Tympner et al
1979
Leber Magen Darm
Tympner et al
1977
Verhand Dtschen Gesellschaft fur I
Medizin
Uskudar et al
2009
Pancreas
Valentini et al
1981
Endoscopy
Varghese et al
2002
Clin Radiol
Wang et al
2013
WJG
Wierzbicka-Paczos
et al
1998
Gastroenterologia Polska
Wierzbicka-Paczos
et al
1999
Polski Merk Lek
Will et al
2010
Ultraschall Med
Zaheer et al
2014
Eur J Rad
Bian et al
2014
Chin J Radiol
Braganza et al
1978
Clin Radiol
Gillams et al
2007
Eur J Rad
Helmberger et al
2000
RoFo
Hernandez Garces
et al
2004
J Pancreas
Ho et al
2006
Clin Gastroenterol Hep
Kalmar et al
1984
Southern Medical J
Kalmin et al
2011
Can J Gastroenterol
Kaufman et al
1989
GIE
Kumon et al
2012
GIE
Manfredi et al
1998
La Rad Medica
Novotny et al
2000
Bratisl Lek Listy
Ponette et al
1976
Acta Gastro-Enterol Belgica
Sanyal et al
2012
AJR
Yoshimoto et al
1980
Jap J Gastroenterol
Grossjohann et al
2010
Scand J Gastroenterol
Sood et al
1992
Indian J Gastroenterol
Zhi et al
2002
Chin J Digestive Dis
Zhong et al
2003
WJG
Ainsworth et al
2003
Endoscopy
Bastid et al
1995
J d'Echographie et de Med par Ultr
Campisi et al
2009
Clin Radiol
Dancygier et al
1986
Scand J Gastroenterol
Giday et al
2011
J Gastr Hep
Guarita et al
1982
AMB
Guo et al
2003
Chin J Digestive Dis
Kahl et al
2002
GIE
Kim et al
2001
AJR
Kolmannskog et al
1981
Acta Radiologica
Lackner et al
1980
RoFo
Lawson
1978
Radiology
Manfredi
2002
Radiology
Mao et al
2011
WCJD
Nakashio
1992
Acta medica
Noguchi et al
1985
Gastroenterolog Endoscopy
Propp
2011
Vestnik khirurgii imeni
Rossi et al
1996
Giorn Ital End Dig
Sahel et al
1976
Acta Endoscopica
Seicean et al
2010
Ultraschall Med
Sildiroglu
1985
Rontgenpraxis
Singh et al
1993
Indian J Rad Imag
Sivak et al
1986
Scand J Gastroenterol
Stabile Ianora et al
2013
Recenti Prog Med
Stevens et al
2008
Dig Dis Sci 3831 Eur Radiol (2017) 27:3820–3844 Author
Year
Journal
Reason for exclusion
Tirkes et al
2016
J MRI
No diagnostic values for CP
Trikudanathan et al
2015
Am J Gastroenterol
No diagnostic values for CP
Tripathi et al
2002
Indian J Gastroenterol
No diagnostic values for CP
Tympner et al
1979
Leber Magen Darm
No diagnostic values for CP
Tympner et al
1977
Verhand Dtschen Gesellschaft fur Innere
Medizin
No diagnostic values for CP
Uskudar et al
2009
Pancreas
No diagnostic values for CP
Valentini et al
1981
Endoscopy
No diagnostic values for CP
Varghese et al
2002
Clin Radiol
No diagnostic values for CP
Wang et al
2013
WJG
No diagnostic values for CP
Wierzbicka-Paczos
et al
1998
Gastroenterologia Polska
No diagnostic values for CP
Wierzbicka-Paczos
et al
1999
Polski Merk Lek
No diagnostic values for CP
Will et al
2010
Ultraschall Med
No diagnostic values for CP
Zaheer et al
2014
Eur J Rad
No diagnostic values for CP
Bian et al
2014
Chin J Radiol
No reference standard
Braganza et al
1978
Clin Radiol
No reference standard
Gillams et al
2007
Eur J Rad
No reference standard
Helmberger et al
2000
RoFo
No reference standard
Hernandez Garces
et al
2004
J Pancreas
No reference standard
Ho et al
2006
Clin Gastroenterol Hep
No reference standard
Kalmar et al
1984
Southern Medical J
No reference standard
Kalmin et al
2011
Can J Gastroenterol
No reference standard
Kaufman et al
1989
GIE
No reference standard
Kumon et al
2012
GIE
No reference standard
Manfredi et al
1998
La Rad Medica
No reference standard
Novotny et al
2000
Bratisl Lek Listy
No reference standard
Ponette et al
1976
Acta Gastro-Enterol Belgica
No reference standard
Sanyal et al
2012
AJR
No reference standard
Yoshimoto et al
1980
Jap J Gastroenterol
No reference standard
Grossjohann et al
2010
Scand J Gastroenterol
Not enough patients
Sood et al
1992
Indian J Gastroenterol
Not enough patients
Zhi et al
2002
Chin J Digestive Dis
Not enough patients
Zhong et al
2003
WJG
Not enough patients
Ainsworth et al
2003
Endoscopy
Only sensitivity reported
Bastid et al
1995
J d'Echographie et de Med par Ultrasons
Only sensitivity reported
Campisi et al
2009
Clin Radiol
Only sensitivity reported
Dancygier et al
1986
Scand J Gastroenterol
Only sensitivity reported
Giday et al
2011
J Gastr Hep
Only sensitivity reported
Guarita et al
1982
AMB
Only sensitivity reported
Guo et al
2003
Chin J Digestive Dis
Only sensitivity reported
Kahl et al
2002
GIE
Only sensitivity reported
Kim et al
2001
AJR
Only sensitivity reported
Kolmannskog et al
1981
Acta Radiologica
Only sensitivity reported
Lackner et al
1980
RoFo
Only sensitivity reported
Lawson
1978
Radiology
Only sensitivity reported
Manfredi
2002
Radiology
Only sensitivity reported
Mao et al
2011
WCJD
Only sensitivity reported
Nakashio
1992
Acta medica
Only sensitivity reported
Noguchi et al
1985
Gastroenterolog Endoscopy
Only sensitivity reported
Propp
2011
Vestnik khirurgii imeni
Only sensitivity reported
Rossi et al
1996
Giorn Ital End Dig
Only sensitivity reported
Sahel et al
1976
Acta Endoscopica
Only sensitivity reported
Seicean et al
2010
Ultraschall Med
Only sensitivity reported
Sildiroglu
1985
Rontgenpraxis
Only sensitivity reported
Singh et al
1993
Indian J Rad Imag
Only sensitivity reported
Sivak et al
1986
Scand J Gastroenterol
Only sensitivity reported Eur Radiol (2017) 27:3820–3844 3832 Author
Year
Journal
Reason for exclusion
Stevens et al
2010
Dig Dis Sci
Only sensitivity reported
Triller et al
1983
Computertomographie
Only sensitivity reported
Uchida et al
1997
Jap J Clin Radiology
Only sensitivity reported
Vitale et al
2009
The Am Surgeon
Only sensitivity reported
Wang et al
2009
J Gastr Hep
Only sensitivity reported
Wu et al
2006
World Chin J Dig
Only sensitivity reported
Yanling et al
2001
Chinese J Gastroenterol
Only sensitivity reported
Zhou et al
1993
Zhonghua nei ke za zhi
Only sensitivity reported
Aithal et al
2002
GIE
Other disease
Doust et al
1976
Radiology
Other disease
Engjom et al
2015
Scan J Gastroenterol
Other disease
Huang et al
2009
Acad J Sec Mil Med University
Other disease
Kushnir et al
2011
GIE
Other disease
Lai et al
2004
Endoscopy
Other disease
Leblanc et al
2014
Pancreas
Other disease
Matos et al
2001
GIE
Other disease
Mosler et al
2012
Dig Dis Sci
Other disease
Novis et al
2010
Rev Colegio Brasileiro Cirurg
Other disease
Rana et al
2012
J Gastr Hep
Other disease
Ranney et al
2012
GIE
Other disease
Sainani et al
2015
Pancreas
Other disease
Soto et al
2005
Radiology
Other disease
Akisik et al
2009
Radiology
Other imaging modality
Cherian et al
2010
HPB Surgery
Other imaging modality
Glaser et al
1994
Int J Pancreatology
Other imaging modality
Glaser et al
1989
Scand J Gastroenterol
Other imaging modality
Glaser et al
1985
Ultraschall Med
Other imaging modality
Hocke et al
2007
Pancreas
Other imaging modality
Kumon et al
2010
GIE
Other imaging modality
Saftoiu et al
2008
GIE
Other imaging modality
Sreenarasimhaiah
2008
J Clin Gastroenterol
Other imaging modality
Tummula et al
2013
Clin Transl Gastroenterol
Other imaging modality
Uehara et al
2011
J Gastr Hep
Other imaging modality
Abdalla et al
2012
Gastroenterolgy
Other type of article
Arsac et al
1981
Med Chirurgie Digest
Other type of article
Ashida et al
2011
J Gastr Hep
Other type of article
Chvatalova et al
2012
Pancreatology
Other type of article
Czako et al
2007
J Gastroenterol
Other type of article
Gupta et al
2013
JIMSA
Other type of article
Heverhagen et al
2007
RoFo
Other type of article
Kasugai et al
1982
Stomach and intestine
Other type of article
Kent et al
2008
Pancreas
Other type of article
Markwardt et al
1980
Radiologia Diagn
Other type of article
Munoz et al
2010
Rev Med de Chile
Other type of article
Musunuri et al
2015
Ind J Gastroenterol
Other type of article
Quinn et al
2012
Gut
Other type of article
Romagnuolo et al
2012
GIE
Other type of article
Sherman et al
2012
GIE
Other type of article
Shibukawa et al
2015
Dig Endos
Other type of article
Stevens et al
2008
Pancreas
Other type of article
Takahashi et al
2014
AJR
Other type of article
Trus et al
1998
Probl Gen Surg
Other type of article
Vadrot et al
1981
Med Chirurgie Digest
Other type of article
Zaruba et al
2012
Pancreatology
Other type of article
Zhang et al
2011
J Gastr Hep
Other type of article Table 6 (continued) 3833 Eur Radiol (2017) 27:3820–3844 Table 7
QUADAS-2 characteristics for each study
Study
Bias
Applicability
Patient selection Index test Reference standard Flow and timing Patient selection Index test Reference stan
Adamek et al
Low
Low
Low
Low
Unclear
Unclear
Low
Albashir et al
Low
Low
Low
Low
Low
Low
Low
Alcaraz et al
Low
Low
Low
Low
High
Unclear
Low
Balci et al
Low
Low
Unclear
Low
Low
Low
Unclear
Bolog et al
Low
Unclear
Low
Low
High
Unclear
Low
Brand et al
Low
Low
Low
High
High
Low
Low
Buscail et al
Low
Unclear
Low
Low
High
Unclear
Low
Catalano et al
Unclear
Low
Unclear
Low
Low
Low
Low
Chong et al
Low
Low
Low
Low
Low
Low
Low
Conwell et al
Low
Low
High
Low
Low
Low
Unclear
Dramaix et al
Low
Low
Low
Low
Low
Unclear
Low
Fusari et al
Unclear
Low
Low
Low
High
Low
Low
Gebel et al
Low
Low
Low
High
Low
Unclear
Low
Giovannini et al
Unclear
Unclear
Low
Low
High
Unclear
Unclear
Glasbrenner et al
Low
Low
Low
Low
High
Low
Low
Gmelin et al
Low
Low
Low
Low
Low
High
Unclear
Hellerhoff et al
Low
Low
Low
Low
Low
Low
Low
Imdahl et al
Low
Low
Unclear
Low
Low
Unclear
Low
Kremer et al
High
Unclear
Unclear
High
High
Unclear
Low
Lammer et al
Low
Low
Unclear
Low
Low
Unclear
Unclear
Lawson et al
Low
Low
Unclear
Low
Low
Low
Unclear
Lees et al
Low
Low
Low
High
Low
High
Low
Lin et al
High
Unclear
Low
Low
Low
Unclear
Low
Nattermann et al
Unclear
Low
Low
Low
High
Unclear
Low
Pamos et al
Low
Low
Low
Low
High
Unclear
Low
Parsi et al
Low
Low
Low
Low
Low
Low
Low
Pistolesi et al
Unclear
Low
Low
Low
Low
Low
Low
Pungpapong et al
Low
Low
Low
Low
Low
Low
Low
Pungpapong et al
Low
Unclear
Unclear
Low
Low
Low
Low
Rudowicz Pietr-uszewska et al Low
Unclear
Low
Low
High
Unclear
Unclear
Sai et al
High
Low
Low
Low
Low
Low
Low
Savarino et al
Unclear
Low
Low
Low
Low
Low
Low
Scarabino et al
Low
Unclear
Unclear
Low
High
Unclear
Unclear
Schlaudraff et al
Low
Unclear
Low
Low
Low
Low
Low
Stevens et al
Low
Low
Unclear
Low
Low
Low
Unclear
Sverko et al
Unclear
Unclear
Low
Low
Low
Unclear
Low
Swobodnik et al
Low
Low
Low
Low
Low
Low
Low
Tox et al
Low
Unclear
Unclear
Low
Low
Low
Low
Trikudanathan et al
Unclear
Low
Unclear
Low
High
Low
Low
Triller et al
Unclear
Low
Unclear
Low
Unclear
Unclear
Low
Wiersema et al
Unclear
Low
Unclear
Low
High
Low
Unclear
Zhang et al
High
Unclear
High
Low
Low
Unclear
High
Zuccaro et al
Unclear
Low
Unclear
Low
Low
Low
Unclear Table 7
QUADAS-2 characteristics for each study Eur Radiol (2017) 27:3820–3844 3834 EUS
Outcome
№of studies
(№of patients)
Study design
Factors that may decrease quality of evidence
Effect per 1000
patients tested
Quality of
evidence
Risk of bias
Indirectness
Inconsistency
Imprecision
Publication bias
Pre-test probability
of 47.2%
True positives
16 (1249)
Cohort & case-control
Serious a
Serious b
Very serious c
Very serious d
NA
387 (335 to 425)
⨁◯◯◯VERY
LOW
False negatives
85 (47 to 137)
True negatives
16 (1249)
Cohort & case-control
Serious a
Serious b
Serious c
Serious d
NA
480 (438 to 502)
⨁◯◯◯VERY
LOW
False positives
48 (26 to 90)
ERCP
Outcome
№of studies
(№of patients)
Study design
Factors that may decrease quality of evidence
Effect per 1000
patients tested
Quality of
evidence
Risk of bias
Indirectness
Inconsistency
Imprecision
Publication bias
Pre-test probability
of 42.6%
True positives
11 (742)
Cohort & case-control
Not serious e
Serious f
Serious g
Serious h
NA
349 (324 to 371)
⨁◯◯◯
Very low
False negatives
77 (55 to 102)
True negatives
11 (742)
Cohort & case-control
Not serious e
Serious f
Serious g
Serious h
NA
540 (499 to 563)
⨁◯◯◯
Very low
False positives
34 (11 to 75)
MRCP
Outcome
№of studies
(№of patients)
Study design
Factors that may decrease quality of evidence
Effect per 1000
patients tested
Quality of
evidence
Risk of bias
Indirectness
Inconsistency
Imprecision
Publication bias
Pre-test probability
of 28.9%
True positives
14 (933)
Cohort & case-control-
type studies
Serious i
Serious j
Serious k
Very serious l
NA
225 (199 to 246)
⨁◯◯◯
Very low
False negatives
64 (43 to 90)
True negatives
14 (933)
Cohort & case-control-
type studies
Serious i
Serious j
Serious k
Not serious l
NA
683 (640 to 697)
⨁◯◯◯
Very low
False positives
28 (14 to 71)
CT
Outcome
№of studies
(№of patients)
Study design
Factors that may decrease quality of evidence
Effect per 1000
patients tested
Quality of
evidence
Risk of bias
Indirectness
Inconsistency
Imprecision
Publication bias
Pre-test probability
of 38.4%
True positives
10 (700)
Cohort & case-control
Serious m
Serious n
Serious o
Very serious p
NA
288 (253 to 319)
⨁◯◯◯
Very low
False negatives
96 (65 to 131)
True negatives
10 (700)
Cohort & case-control
Serious m
Serious n
Serious o
Serious p
NA
561 (499 to 591)
⨁◯◯◯
Very low
False positives
55 (25 to 117)
US
Outcome
№of studies
(№of patients)
Study design
Factors that may decrease quality of evidence
Effect per 1000
patients tested
Quality of
evidence
Risk of bias
Indirectness
Inconsistency
Imprecision
Publication bias
pre-test probability
of 25.7% Eur Radiol (2017) 27:3820–3844 3835 nued)
10 (1005)
Cohort & case-control
Serious q
Serious r
Serious s
Very serious t
NA
172 (136 to 200)
⨁◯◯◯
Very low
85 (57 to 121)
10 (1005)
Cohort & case-control
Serious q
Serious r
Very serious s
Serious t
NA
728 (661 to 743)
⨁◯◯◯
Very low
15 (0 to 82)
e
ased on QUADAS-2 risk of bias; 7 studies not serious, 9 studies serious
ased on QUADAS-2 applicability; 7 studies not serious, 9 studies serious
based on heterogeneity and visual inspection CIs
ased on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
ADAS-2 risk of bias: 8 studies not serious, 3 studies serious
ADAS-2 applicability: 6 studies not serious, 5 studies serious
rogeneity and visual inspection CIs
y numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
ased on QUADAS-2 risk of bias; 7 studies not serious, 5 studies serious, 1 study very serious
ased on QUADAS-2 applicability; 6 studies not serious, 8 studies serious
based on heterogeneity and visual inspection CIs
ased on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
based on QUADAS-2 risk of bias; 5 studies not serious, 5 studies serious
ased on QUADAS-2 applicability; 6 studies not serious, 4 studies serious
based on heterogeneity and visual inspection CIs
ased on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
based on QUADAS-2 risk of bias; 6 studies not serious, 3 studies serious, 1 study very serious
ased on QUADAS-2 applicability; 5 studies not serious, 5 studies serious
based on heterogeneity and visual inspection CIs
ased on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
ur Radiol (2017) 27:3820–3844
383 ued)
10 (1005)
Cohort & case-control
Serious q
Serious r
Serious s
Very serious t
NA
172 (136 to 200)
⨁◯◯◯
Very low
85 (57 to 121)
10 (1005)
Cohort & case-control
Serious q
Serious r
Very serious s
Serious t
NA
728 (661 to 743)
⨁◯◯◯
Very low
15 (0 to 82)
ased on QUADAS-2 risk of bias; 7 studies not serious, 9 studies serious
ased on QUADAS-2 applicability; 7 studies not serious, 9 studies serious
based on heterogeneity and visual inspection CIs
sed on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
DAS-2 risk of bias: 8 studies not serious, 3 studies serious
DAS-2 applicability: 6 studies not serious, 5 studies serious
ogeneity and visual inspection CIs
numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
sed on QUADAS-2 risk of bias; 7 studies not serious, 5 studies serious, 1 study very serious
sed on QUADAS-2 applicability; 6 studies not serious, 8 studies serious
based on heterogeneity and visual inspection CIs
sed on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
ased on QUADAS-2 risk of bias; 5 studies not serious, 5 studies serious
ased on QUADAS-2 applicability; 6 studies not serious, 4 studies serious
based on heterogeneity and visual inspection CIs
sed on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
ased on QUADAS-2 risk of bias; 6 studies not serious, 3 studies serious, 1 study very serious
sed on QUADAS-2 applicability; 5 studies not serious, 5 studies serious
based on heterogeneity and visual inspection CIs
sed on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
(
) Table 8 (continued)
True positives
10 (1005)
Cohort & case-control
Serious q
Serious r
Serious s
Very serious t
NA
False negatives
True negatives
10 (1005)
Cohort & case-control
Serious q
Serious r
Very serious s
Serious t
NA
False positives
NA not available
a Risk of bias: based on QUADAS-2 risk of bias; 7 studies not serious, 9 studies serious
b Indirectness: based on QUADAS-2 applicability; 7 studies not serious, 9 studies serious
c Inconsistency: based on heterogeneity and visual inspection CIs
d Imprecision: based on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
e Based on QUADAS-2 risk of bias: 8 studies not serious, 3 studies serious
f Based on QUADAS-2 applicability: 6 studies not serious, 5 studies serious
g Based on heterogeneity and visual inspection CIs
h Based on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
i Risk of bias: based on QUADAS-2 risk of bias; 7 studies not serious, 5 studies serious, 1 study very serious
j Indirectness: based on QUADAS-2 applicability; 6 studies not serious, 8 studies serious
k Inconsistency: based on heterogeneity and visual inspection CIs
l Imprecision: based on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
m Risk of bias: based on QUADAS-2 risk of bias; 5 studies not serious, 5 studies serious
n Indirectness: based on QUADAS-2 applicability; 6 studies not serious, 4 studies serious
o Inconsistency: based on heterogeneity and visual inspection CIs
p Imprecision: based on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious)
q Risk of bias: based on QUADAS-2 risk of bias; 6 studies not serious, 3 studies serious, 1 study very serious
r Indirectness: based on QUADAS-2 applicability; 5 studies not serious, 5 studies serious
s Inconsistency: based on heterogeneity and visual inspection CIs
t Imprecision: based on study numbers and CIs of summary estimate (CIs 0–10 = not serious, 11–15 = serious, more than 15 = very serious) Eur Radiol (2017) 27:3820–3844 3836 Table 9
GRADE characteristics for each study
Modality
Name first author
Risk of bias
Indirectness
Inconsistency
Imprecision
Publication bias
EUS
Albashir et al
Low
Low
Sensitivity: very serious
Specificity: serious
Sensitivity: very serious
Specificity: serious
Not assessed
Brand et al
Serious
Serious
Buscail et al
Serious
Serious
Catalano et al
Low
Low
Chong et al
Low
Low
Conwell et al
Serious
Serious
Giovannini et al
Serious
Serious
Glasbrenner et al
Low
Low
Lin et al
Serious
Serious
Nattermann et al
Low
Serious
Pungpapong et al
Low
Low
Pungpapong et al
Low
Low
Stevens et al
Serious
Serious
Tox et al
Serious
Low
Trikudanathan et al
Serious
Serious
Wiersema et al
Serious
Serious
ERCP
Adamek et al
Low
Low
Sensitivity: serious
Specificity: serious
Sensitivity: serious
Specificity: serious
Not assessed
Buscail et al
Low
Serious
Gebel et al
Low
Low
Glasbrenner et al
Low
Low
Gmelin et al
Low
Serious
Lammer et al
Serious
Serious
Lawson et al
Low
Low
Parsi et al
Low
Low
Scarabino et al
Serious
Serious
Swobodnik et al
Low
Low
Triller et al
Serious
Serious
MRCP
Adamek et al
Low
Low
Sensitivity: serious
Specificity: serious
Sensitivity: very serious
Specificity: not serious
Not assessed
Alcaraz et al
Low
Serious
Balci et al
Serious
Serious
Bolog et al
Serious
Serious
Fusari et al
Low
Low
Hellerhoff et al
Low
Low
Pamos et al
Low
Serious
Pungpapong et al
Low
Low
Rudowicz-Pietruszewska
Serious
Serious
Sai et al
Serious
Low
Schlaudraff et al
Low
Low
Sverko et al
Serious
Serious
Zhang et al
Very serious
Serious
Zuccaro et al
Serious
Serious
CT
Buscail et al
Low
Serious
Sensitivity: serious
Specificity: serious
Sensitivity: very serious
Specificity: serious
Not assessed
Dramaix et al
Low
Low
Fusari et al
Low
Low
Gmelin et al
Low
Serious
Imdahl et al
Serious
Low
Lammer et al
Serious
Serious
Pistolesi et al
Low
Low
Savarino et al
Serious
Low
Scarabino et al
Serious
Serious
Swobodnik et al
Low
Low
US
Buscail et al
Low
Serious
Sensitivity: serious
Specificity: very serious
Sensitivity: very serious
Specificity: serious
Not assessed
Dramaix et al
Low
Low
Gebel et al
Low
Low
Gmelin et al
Low
Serious
Kremer et al
Very serious
Serious
Lawson et al
Low
Low
Lees et al
Serious
Low
Lin et al
Serious
Serious
Scarabino et al
Serious
Serious
Swobodnik et al
Low
Low Table 9
GRADE characteristics for each study Sensitivity: serious
Specificity: serious Sensitivity: very serious
Specificity: not serious
Not assessed Sensitivity: serious
Specificity: serious Eur Radiol (2017) 27:3820–3844 3837 Table 10
Diagnostic characteristics for each study
Study
Sensitivity
Specificity
Accuracy
PPV
NPV
TP
TN
FP
FN
Adamek et al
MRCP: 88%,
ERCP: 90%
MRCP: 94%. EUS [All Fields] ERCP:
91%
MRCP: 91% ERCP:
90%
MRCP: 93%,
ERCP:90%
MRCP: 90%, ERCP:
91%
MRCP:50
ERCP: 51
MRCP: 63
ERCP: 61
MRCP: 4
ERCP: 6
MRCP: 7
ERCP: 6
Albashir et al
84%
100%
87%
100%
57%
16
4
0
3
Alcaraz et al
50%
99%
94%
80%
95%
4
72
1
4
Balci et al
82%
63%
70%
56%
86%
9
12
7
2
Bolog et al
90%
98%
95%
90%
98%
14
86
2
1
Brand et al
42%
96%
84%
71%
86%
10
87
4
14
Buscail et al
US: 58%,CT: 75%,
ERCP: 74%,
EUS: 88%
US: 75%, CT: 95%,
ERCP: 100%,
EUS: 100%
US: 65%, CT: 81%,
ERCP: 82%,
EUS: 92%
US: 87%, CT: 97%,
ERCP: 100%,
EUS: 100%
US: 44%, CT: 61%,
ERCP: 62%,
EUS: 78%
US: 26, CT: 33,
ERC: 33,
EUS: 39
US: 14, CT: 17,
ERCP: 18,
EUS: 18
US: 4, CT: 1,
ERCP: 0,
EUS: 0
US: 18, CT: 11,
ERCP: 11,
EUS: 5
Catalano et al
84%
98%
91%
97%
87%
32
41
1
6
Chong et al
83%
80%
83%
98%
69%
53
5
1
11
Conwell et al
26%
100%
50%
100%
39%
10
18
0
28
Dramaix et al
CT: 60%, US: 60%
CT: 100% US: 95%
CT: 86% US: 82%
CT: 100%, US: 90%
CT: 76%, US: 76%
CT: 11, US: 11
CT: 32, US: 30
CT: 0, US: 2
CT: 7, US: 7
Fusari et al
CT: 88%, MRI: 88% CT: 100%,
MRI:100%,
CT: 98%, MRI: 98% CT: 100%, MRI:
100%
CT: 97%, MRI: 97% MRI: 7, CT: 7
MRI: 32, CT: 32 MRI: 0, CT: 0
MRI: 1, CT: 1
Gebel et al
US: 82%, ERP: 56% US: 97%, ERP: 97%
US: 91%, ERP: 82% US:95%, ERP: 90%
US: 89%, ERP: 80% US: 18, ERP: 9
US: 33, ERP: 28 US: 1, ERP: 1
US: 4, ERP: 7
Giovannini et al
94%
56%
81%
80%
83%
16
5
4
1
Glasbrenner et al
EUS: 93%, ERCP:
88%
EUS: 78%, ERCP:
82%
EUS: 85%, ERCP:
85%
EUS: 79%, ERCP:
82%
EUS: 92%, ERCP:
88%
EUS: 38,
ERCP: 36
EUS: 34, ERCP:
36
EUS: 10,
ERCP: 8
EUS: 3, ERCP:
5
Gmelin et al
US: 68%, CT: 84%,
ERCP: 89%
US: 100%, CT: 91%,
ERCP: 91%
US: 85%, CT: 89%,
ERCP: 90%
US: 100%, CT: 89%,
ERCP: 89%
US: 79%, CT: 87%,
ERCP: 91%
US: 13, CT: 16,
ERCP: 17
US: 22, CT: 20,
ERCP: 20
US: 0, CT: 2,
ERCP: 2
US: 6, CT: 3. EUS [All Fields] ERCP: 2
Hellerhoff et al
MRI: 77%, sMRI:
89%
MRI: 100%,
sMRI:100%
MRI 94%, sMRI:
97%
MRI: 100%, sMRI:
100%
MRI: 92%, sMRI:
96%
MRI: 20, sMRI:
23
MRI: 69, sMRI:
69
MRI: 0, sMRI:
0
MRI: 6, sMRI:
3
Imdahl et al
58%
91%
83%
70%
85%
7
33
3
5
Kremer et al
67%
99%
94%
89%
95%
42
378
5
21
Lammer et al
ERCP: 85%, CT:
64%
ERCP: 97%, CT:
85%
ERCP: 93%, CT:
78%
ERCP: 94%, CT:
71%
ERCP: 92%, CT:
81%
ERCP: 33, CT:
25
ERCP: 66, CT:
58
ERCP: 2, CT:
10
ERCP: 6, CT:
14
Lawson et al
US: 38%, ERCP:
73%
US: 100%, ERCP:
98%
US: 79%, ERCP:
98%
US: 100%, ERCP:
95%
US: 75%, ERCP:
87%
US: 10, ERCP:
19
US: 49, ERCP:
48
US: 0, ERCP:
1
US: 16, ERCP:
7
Lees et al
100%
97%
98%
91%
100%
20
76
2
0
Lin et al
US: 86%, EUS:
100%
US: 100%, EUS:
100%
US: 97%, EUS:
100%
US: 100%, EUS:
100%
US: 96%, EUS:
100%
US: 6, EUS: 7
US: 26, EUS: 26 US: 0, EUS: 0
US: 1, EUS: 0
Nattermann et al
98%
57%
75%
65%
97%
50
36
27
1
Pamos et al
80%
100%
98%
100%
97%
4
36
0
1
Parsi et al
71%
91%
77%
94%
59%
17
10
1
7
Pistolesi et al
58%
81%
74%
58%
81%
18
56
13
13
Pungpapong et al
71%
88%
80%
84%
77%
27
36
5
11
Pungpapong et al
EUS: 93%, MRCP:
65%
EUS: 93%, MRCP:
90%
EUS: 93%, MRCP:
80%
EUS: 90%, MRCP:
81%
EUS: 95%, MRCP:
79%
EUS: 37,
MRCP: 26
EUS: 55,
MRCP: 53
EUS: 4,
MRCP: 6
EUS: 3, MRCP:
14
Rudowicz-Pietruszewska
et al
100%
100%
100%
100%
100%
9
79
0
0
Sai et al
60%
79%
68%
77%
60%
10
9
3
6
Savarino et al
90%
59%
76%
73%
83%
53
29
20
6
Scarabino et al
ERCP: 83%, US:
42%, CT: 100%
ERCP: 67%, US:
34%, CT: 70%
ERCP: 70%, US:
35%, CT: 76%
ERCP: 37%, US:
13%, CT: 44%
ERCP: 94%, US:
71%, CT: 100%
ERCP: 10, US:
5, CT: 12
ERCP: 34, US:
17, CT: 36
ERCP: 17,
US: 34, CT:
15
ERCP: 2, US: 7,
CT: 0
Eur Radiol (2017) 27:3820–3844
383 Eur Radiol (2017) 27:3820–3844 3838 (
)
Study
Sensitivity
Specificity
Accuracy
PPV
NPV
TP
TN
FP
FN
Schlaudraff et al
MRCP: 67%,
sMRCP: 73%
MRCP: 93%,
sMRCP: 96%
MRCP: 89%,
sMRCP: 93%
MRCP: 63%,
sMRCP: 78%
MRCP: 95%,
sMRCP: 95%
MRCP: 6,
sMRCP: 7
MRCP: 49,
sMRCP: 51
MRCP: 4,
sMRCP: 2
MRCP: 3,
sMRCP: 2
Stevens et al
Radial: 68%, Linear:
44%
Radial: 95% Linear:
95%
Radial: 84% Linear:
74%
Radial: 90%, Linear:
86%
Radial: 81%, Linear:
71%
28
56
3
13
Sverko et al
79%
93%
86%
92%
82%
11
14
1
3
Swobodnik et al
US: 52%, CT: 74%,
ERCP: 93%
US: 100%, CT: 98%,
ERCP: 100%
US: 84%, CT: 90%,
ERCP: 98%
US: 100%, CT: 95%,
ERCP: 100%
US: 81%, CT: 88%,
ERCP: 96%
US: 14, CT: 20,
ERCP: 25
US: 54, CT: 53,
ERCP: 54
US: 0, CT: 1,
ERCP: 0
US: 13, CT: 7,
ERCP: 2
Tox et al
77%
75%
76%
66%
84%
50
80
26
15
Trikudanathan et al
61%
75%
63%
92%
29%
34
9
3
22
Triller et al
82%
85%
83%
82%
85%
9
11
2
2
Wiersema et al
80%
86%
84%
83%
84%
24
32
5
6
Zhang et al
92%
75%
84%
81%
88%
22
15
5
2
Zuccaro et al
MRCP: 46%,
sMRCP: 46%
MRCP:85%,
sMRCP: 68%
MRCP: 70%,
sMRCP: 59%
MRCP: 68%,
sMRCP: 50%
MRCP: 70%,
sMRCP: 65%
MRCP: 13,
sMRCP: 13
MRCP: 35,
sMRCP: 28
MRCP: 6,
sMRCP: 13
MRCP: 15,
sMRCP: 15
PPV positive predictive value, NPV negative predictive value, TP true positive, TN true negative, FP false positive, FN false negative
Table 11
Imaging characteristics for each study
Magnetic resonance imaging (MRI)
Study
Year Magnetic field
Coil type
Contrast
Secretin
enhancement
Sequence
Scoring criteria
Adamek et al
2000
1.0 T
Body coil
No
No
T2
Size of common bile and pancreatic duct,
the nature and degree of pancreatic
duct obstruction, and accuracy
in diagnosing pathological findings
Alcaraz et al
2000
1.5 T
NA
No
No
T2 (HASTE & RARE)
NA
Balci et al
2006
1.5 T
Four-element quadrature
phased-array surface coil
IV
No
T1, T2
Increased arterial enhancement pattern,
normal gland size and normal ductal
morphology (Cambridge classification)
Bolog et al
2004
1.0 T
Synergy body coil
NA
No
T1, T2
NA
Fusari et al
2010
1.5 T
Phased-array synergy body
coil
Oral
No
T1, T2
1–5 score to identify pancreatic masses
(definite benign = 1, probably
benign = 2 etc.)
Hellerhoff et al
2002
1.5 T
Phased-array synergy
surface coil
Oral
No
T2
Cambridge classification
Pamos et al
1998
1.5 T
Body coil
NA
No
T2
NA
Pungpapong et al
2007
1.5 T
Phased-array surface coil
IV/Oral
No
T1, T2, T2 (HASTE)
Presence of 1 or more of the following
features: main pancreatic duct dilatation
in absence of structural obstruction,
dilated side branches, intraductal stones,
(
) sMRCP: 46%
sMRCP: 68%
sMRCP: 59%
sMRCP: 50%
sMRCP: 65%
sMRCP: 13
sMRCP: 28
sMRCP: 13
sMRCP: 15
PPV positive predictive value, NPV negative predictive value, TP true positive, TN true negative, FP false positive, FN false negative
Table 11
Imaging characteristics for each study
Magnetic resonance imaging (MRI)
Study
Year Magnetic field
Coil type
Contrast
Secretin
enhancement
Sequence
Scoring criteria
Adamek et al
2000
1.0 T
Body coil
No
No
T2
Size of common bile and pancreatic duct,
the nature and degree of pancreatic
duct obstruction, and accuracy
in diagnosing pathological findings
Alcaraz et al
2000
1.5 T
NA
No
No
T2 (HASTE & RARE)
NA
Balci et al
2006
1.5 T
Four-element quadrature
phased-array surface coil
IV
No
T1, T2
Increased arterial enhancement pattern,
normal gland size and normal ductal
morphology (Cambridge classification)
Bolog et al
2004
1.0 T
Synergy body coil
NA
No
T1, T2
NA
Fusari et al
2010
1.5 T
Phased-array synergy body
coil
Oral
No
T1, T2
1–5 score to identify pancreatic masses
(definite benign = 1, probably
benign = 2 etc.)
Hellerhoff et al
2002
1.5 T
Phased-array synergy
surface coil
Oral
No
T2
Cambridge classification
Pamos et al
1998
1.5 T
Body coil
NA
No
T2
NA
Pungpapong et al
2007
1.5 T
Phased-array surface coil
IV/Oral
No
T1, T2, T2 (HASTE)
Presence of 1 or more of the following
features: main pancreatic duct dilatation
in absence of structural obstruction,
dilated side branches, intraductal stones, Study
Sensitivity
Specificity
Accuracy
PPV
NPV
TP
TN
FP
FN
Schlaudraff et al
MRCP: 67%,
sMRCP: 73%
MRCP: 93%,
sMRCP: 96%
MRCP: 89%,
sMRCP: 93%
MRCP: 63%,
sMRCP: 78%
MRCP: 95%,
sMRCP: 95%
MRCP: 6,
sMRCP: 7
MRCP: 49,
sMRCP: 51
MRCP: 4,
sMRCP: 2
MRCP: 3,
sMRCP: 2
Stevens et al
Radial: 68%, Linear:
44%
Radial: 95% Linear:
95%
Radial: 84% Linear:
74%
Radial: 90%, Linear:
86%
Radial: 81%, Linear:
71%
28
56
3
13
Sverko et al
79%
93%
86%
92%
82%
11
14
1
3
Swobodnik et al
US: 52%, CT: 74%,
ERCP: 93%
US: 100%, CT: 98%,
ERCP: 100%
US: 84%, CT: 90%,
ERCP: 98%
US: 100%, CT: 95%,
ERCP: 100%
US: 81%, CT: 88%,
ERCP: 96%
US: 14, CT: 20,
ERCP: 25
US: 54, CT: 53,
ERCP: 54
US: 0, CT: 1,
ERCP: 0
US: 13, CT: 7,
ERCP: 2
Tox et al
77%
75%
76%
66%
84%
50
80
26
15
Trikudanathan et al
61%
75%
63%
92%
29%
34
9
3
22
Triller et al
82%
85%
83%
82%
85%
9
11
2
2
Wiersema et al
80%
86%
84%
83%
84%
24
32
5
6
Zhang et al
92%
75%
84%
81%
88%
22
15
5
2
Zuccaro et al
MRCP: 46%,
sMRCP: 46%
MRCP:85%,
sMRCP: 68%
MRCP: 70%,
sMRCP: 59%
MRCP: 68%,
sMRCP: 50%
MRCP: 70%,
sMRCP: 65%
MRCP: 13,
sMRCP: 13
MRCP: 35,
sMRCP: 28
MRCP: 6,
sMRCP: 13
MRCP: 15,
sMRCP: 15
PPV positive predictive value, NPV negative predictive value, TP true positive, TN true negative, FP false positive, FN false negative Table 10 (continued) Severe CP: atrophy
or diffuse/focal enlargement of the gland,
calcification,
chronic pseudocysts
Ultrasonography (US)
Study
Year
Transducer
Scoring criteria
Buscail et al
1995
NA
NA
Dramaix et al
1980
Unirad/Kretz combison 200
NA
Gebel et al
1985
ADR 2130 Imager 2380 Sonoline 8000
Duct abnormalities
Gmelin et al
1981
Sono fluoroskop 1, unirad model 849
Criteria for PC, CP and normal pancreas
were extracted from literature
Kremer et al
1977
NA
Rettenmaier specified examination technique
Lawson et al
1978
13-mm diameter 3.5 Mhz/ 13 or
19-mm diameter 2.25 Mhz
Identification of a mass, pseudocyst or
generalized glandular enlargement with
abnormal parenchymal echogenicity
ur Radiol (2017) 27:3820–3844
3839 Eur Radiol (2017) 27:3820–3844 3839 Magnetic resonance imaging (MRI)
Study
Year Magnetic field
Coil type
Contrast
Secretin
enhancement
Sequence
Scoring criteria
ductal irregularity, reduced T1 signal
intensity, atrophy of pancreatic
parenchyma and reduced secretory
response to secretin administration
Rudowicz-Pietruszewska
et al
2002
0.5 T
Body coil
NA
No
T2
NA
Schlaudraff et al
2008
1.0 T
Dedicated quadrature torso
phased-array coil
NA
No
T2, T2 (HASTE)
Pancreatic duct stenosis/dilatation, side
branch stenosis/dilatation,
pseudocysts, extrapancreatic abscess. Based on observers’ judgement
Sverko et al
2011
1.0 T
NA
IV
No
T1, T2 (HASTE)
NA
Zhang et al
2003
1.5 T
NA
IV
No
T1
Signal intensity by gadolinium (presence
of SIR less than 1.73 in the arterial phase)
Zuccaro et al
2009
NA
Phased array-torso coil
IV
No
T1, T2, T2 (HASTE)
Mild CP: secretin-induced T2 intensity
significantly reduced; side branch
ectasia, mild ductal dilatation. Moderate
CP: abnormal enhancement
pattern on T1 after gadolinium administration. Severe CP: atrophy
or diffuse/focal enlargement of the gland,
calcification, chronic pseudocysts
Secretin-enhanced magnetic
resonance imaging (sMRI)
Hellerhoff et al
2002
1.5 T
Synerge phased-array
surface coil
IV
Yes
T2
Cambridge classification
Sai et al
2008
1.5 T
Phased-array multi coil
IV
Yes
NA
Cambridge classification
Schlaudraff et al
2008
1.0 T
Dedicated quadrature
phased-array torso coil
NA
Yes
T2, T2 (HASTE)
Pancreatic duct stenosis/dilatation, side
branch stenosis/dilatation,
d
i
b
r Radiol (2017) 27:3820–3844 Coil type
Contrast
Secretin
enhancement
Sequence
Scoring criteria
ductal irregularity, reduced T1 signal
intensity, atrophy of pancreatic
parenchyma and reduced secretory
response to secretin administration
Body coil
NA
No
T2
NA
Dedicated quadrature torso
phased-array coil
NA
No
T2, T2 (HASTE)
Pancreatic duct stenosis/dilatation, side
branch stenosis/dilatation,
pseudocysts, extrapancreatic abscess. Table 10 (continued) Table 11 (continued)
Magnetic resonance imaging (MRI)
Study
Year Magnetic field
Coil type
Contrast
Secretin
enhancement
Sequence
Scoring criteria
ductal irregularity, reduced T1 signal
intensity, atrophy of pancreatic
parenchyma and reduced secretory
response to secretin administration
Rudowicz-Pietruszewska
et al
2002
0.5 T
Body coil
NA
No
T2
NA
Schlaudraff et al
2008
1.0 T
Dedicated quadrature torso
phased-array coil
NA
No
T2, T2 (HASTE)
Pancreatic duct stenosis/dilatation, side
branch stenosis/dilatation,
pseudocysts, extrapancreatic abscess. Based on observers’ judgement
Sverko et al
2011
1.0 T
NA
IV
No
T1, T2 (HASTE)
NA
Zhang et al
2003
1.5 T
NA
IV
No
T1
Signal intensity by gadolinium (presence
of SIR less than 1.73 in the arterial phase)
Zuccaro et al
2009
NA
Phased array-torso coil
IV
No
T1, T2, T2 (HASTE)
Mild CP: secretin-induced T2 intensity
significantly reduced; side branch
ectasia, mild ductal dilatation. Moderate
CP: abnormal enhancement
pattern on T1 after gadolinium administration. Severe CP: atrophy
or diffuse/focal enlargement of the gland,
calcification, chronic pseudocysts
Secretin-enhanced magnetic
resonance imaging (sMRI)
Hellerhoff et al
2002
1.5 T
Synerge phased-array
surface coil
IV
Yes
T2
Cambridge classification
Sai et al
2008
1.5 T
Phased-array multi coil
IV
Yes
NA
Cambridge classification
Schlaudraff et al
2008
1.0 T
Dedicated quadrature
phased-array torso coil
NA
Yes
T2, T2 (HASTE)
Pancreatic duct stenosis/dilatation, side
branch stenosis/dilatation,
pseudocysts, extrapancreatic abscess. Based on observers’ judgement
Zuccaro et al
2009
NA
Phased-array torso coil
IV
Yes
T1, T2, T2 (HASTE)
Mild CP: secretin-induced T2 intensity significantly
reduced; side branch ectasia, mild ductal dilatation. Moderate CP: abnormal enhancement pattern on T1
after gadolinium administration. Table 10 (continued) Based on observers’ judgement
NA
IV
No
T1, T2 (HASTE)
NA
NA
IV
No
T1
Signal intensity by gadolinium (presence
of SIR less than 1.73 in the arterial phase)
Phased array-torso coil
IV
No
T1, T2, T2 (HASTE)
Mild CP: secretin-induced T2 intensity
significantly reduced; side branch
ectasia, mild ductal dilatation. Moderate
CP: abnormal enhancement
pattern on T1 after gadolinium administration. Severe CP: atrophy
or diffuse/focal enlargement of the gland,
calcification, chronic pseudocysts
Synerge phased-array
surface coil
IV
Yes
T2
Cambridge classification
Phased-array multi coil
IV
Yes
NA
Cambridge classification
Dedicated quadrature
phased-array torso coil
NA
Yes
T2, T2 (HASTE)
Pancreatic duct stenosis/dilatation, side
branch stenosis/dilatation,
pseudocysts, extrapancreatic abscess. Based on observers’ judgement
Phased-array torso coil
IV
Yes
T1, T2, T2 (HASTE)
Mild CP: secretin-induced T2 intensity significantly
reduced; side branch ectasia, mild ductal dilatation. Moderate CP: abnormal enhancement pattern on T1
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or diffuse/focal enlargement of the gland,
calcification,
chronic pseudocysts
Scoring criteria
NA
NA
Duct abnormalities
Criteria for PC, CP and normal pancreas
were extracted from literature
Rettenmaier specified examination technique
Identification of a mass, pseudocyst or
generalized glandular enlargement with
abnormal parenchymal echogenicity Scoring criteria
NA
NA
Duct abnormalities
Criteria for PC, CP and normal pancreas
were extracted from literature
Rettenmaier specified examination technique
Identification of a mass, pseudocyst or
generalized glandular enlargement with
abnormal parenchymal echogenicity Eur Radiol (2017) 27:3820–3844 3840 Magnetic resonance imaging (MRI)
Study
Year Magnetic field
Coil type
Contrast
Secretin
enhancement
Sequence
Scoring criteria
Lees et al
1979
2.5 Mhz
Appearance of pancreatic parenchyma and
duct system/size and shape of
the pancreas and from previous reports
Lin et al
1989
SAL-90A 3.75 Mhz
NA
Scarabino et al
1989
NA
NA
Swobodnik et al
1983
Siemens imager 2300 linear array
Organ enlarged or atrophic dense structure,
areas of scars or calcification
(more echogenic), sonolucent areas
only during acute inflammation,
dilatation of the pancreatic duct system,
symmetric contours, no smooth outlines
Computed tomography (CT)
Study
Year
Scanner
Contrast
Scoring criteria
Buscail et al
1995
NA
NA
NA
Dramaix et al
1980
OHIO nuclear - Delta Slan 50FS
Oral/IV
NA
Fusari et al
2010
Marconi MX8000 (four-detector row)
IV
1–5 score to identify pancreatic masses
(definite benign = 1, probably benign = 2 etc.)
Gmelin et al
1981
NA
NA
Criteria for PC, CP and normal pancreas
were extracted from literature
Imdahl et al
1999
Somatom Plus 4 helical scanner
IV
NA
Lammer et al
1980
EMI-5005
Oral
3 stadia typical for CP
Pistolesi et al
1981
Ohio-Nuclear Delta 50 scanner
NA
Overall enlargement of the pancreas or calcifications
Savarino et al
1980
EMI-5005
Oral
Parenchymal atrophy, pancreatic
calcifications, pseudocysts or abscesses
Scarabino et al
1989
NA
NA
NA
Swobodnik et al
1983
General Electric CT-T8800
Oral/IV
Atrophy of the organ (during acute inflammation:
segmental enlargement)
during acute phase; segments without clear
outlines, cysts or calcifications and dense structure
Endoscopic ultrasonography (EUS)
Study
Year
Scanner
Transducer
MHz
Scoring criteria
Albashir et al
2010
NA
NA
NA
9 features; >4 diagnostic for CP
Brand et al
2000
Olympus GF-UM 3/GF-UM
20/GF-UM 200
Radial
NA
Own criteria (increased parenchymal lobulations,
calcification and/or ductal changes or focal lesion)
Buscail et al
1995
Olympus EU-M3
NA
7.5/12
MHz
NA
Catalano et al
1998
Olympus EU-M3/EU-M20
NA
7.5/12
MHz
Wiersema criteria (11 features), own classification system
Chong et al
2007
Olympus EU-M20/GF-UM130/GF-
UM160/GF-UC30P/GF-
UC140P/GF-UCT140
Radial
NA
9 features; >3 diagnostic for CP
Conwell et al
2007
NA
NA
NA
9 features; >3 diagnostic for CP
Giovannini et al
1994
Pentax FG-32-UA
Linear
NA
NA
Glasbrenner et al
2000
Olympus EU-M20
Radial
7.5/12
MHz
Wiersema criteria (11 features)
Lin et al
1989
Olympus GF-EUM 2/GF-UM2
Radial
7.5 MHz
NA
Nattermann et al
1993
Olympus GF-UM-3/EU-M3
NA
7.5/12
MHz
NA
Pungpapong et al
2007
Olympus GF-UE160-AL5/GF-UC140P
Radial & linear
NA
MST criteria; >4 features diagnostic for CP
Pungpapong et al
2007
Olympus GF-UC140P/ UCT140-AL5
Linear
7.5 MHz
MST criteria; >4 features diagnostic for CP Eur Radiol (2017) 27:3820–3844 3841 ab e
(co t ued)
Magnetic resonance imaging (MRI)
Study
Year Magnetic field
Coil type
Contrast
Secretin
enhancement
Sequence
Scoring criteria
Stevens et al
2009
Olympus GF-UM-130/GF-UE-160/GF-
UC-160P-OL5
Radial & linear
NA
9 features; >4 diagnostic for CP
Tox et al
2007
Olympus GF-UM20, Pentax
EG-3620-UR/EG-3830-UT
NA
NA
Own criteria
Trikudanathan
2016
Olympus
Linear
7.5 MHz
Wiersema criteria (11 features) >4 is CP
Wiersema et al
1993
Olympus EU-M3/EU-M20
NA
NA
Wiersema criteria (11 features) >3 is CP
Endoscopic retrograde
cholangiopancreatography (ERCP)
Study
Year
Technical features
Scoring criteria
Adamek et al
2000
NA
NA
Buscail et al
1995
NA
Own criteria (normal/moderate changes
(3 abnormal side branches and normal main
duct)/marked changes (side and
main duct abnormalities))
Gebel et al
1995
NA
Deyhle criteria
Glasbrenner et al
2000
Olympus
Cambridge classification
Gmelin et al
1981
NA
Criteria according to references
Lammer et al
1980
Olympus JFB
Loffler criteria
Lawson et al
1978
NA
Criteria according to references
Parsi et al
2008
NA
Cambridge classification
Scarabino et al
1989
NA
NA
Swobodnik et al
1983
Olympus JFB-2/3
Own criteria (variation in diameter of the
main duct in the whole organ
(exception: segmental pancreatitis),
cystic dilatation of side branches,
kinking of the duct stones in canalicular
structures, distension of the main duct)
Triller et al
1975
NA
NA 3842 Eur Radiol (2017) 27:3820–3844 test, and histology: correlation in chronic pancreatitis. Table 10 (continued) Am J
Gastroenterol 105:2498–2503 Open Access This article is distributed under the terms of the Creative
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distribution, and reproduction in any medium, provided you give
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The Impact of Financial Crises on the Poor
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Johan Rewilak
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Journal of International Development
J. Int. Dev. (2017)
Published online in Wiley Online Library
(wileyonlinelibrary.com) DOI: 10.1002/jid.3334 Journal of International Development
J. Int. Dev. (2017)
Published online in Wiley Online Library
(wileyonlinelibrary.com) DOI: 10.1002/jid.3334 Journal of International Development
J. Int. Dev. (2017)
Published online in Wiley Online Library
(wileyonlinelibrary.com) DOI: 10.1002/jid.3334 Journal of International Development
J. Int. Dev. (2017)
Published online in Wiley Online Library
(wileyonlinelibrary.com) DOI: 10.1002/jid.3334 THE IMPACT OF FINANCIAL CRISES ON
THE POOR JOHAN REWILAK*
Aston Business School, Aston University, Birmingham, UK Abstract: Financial crises have detrimental impacts on the economy via depressed economic growth
and rising unemployment, however, their impact on the poorest in society is relatively under-
researched. This paper investigates the impact of three different types of financial crises on the income
of the poor. Using a variety of estimation techniques and controlling for a lagged dependent variable,
the results suggest that currency crises are the most harmful to the poor, followed by banking crises. Debt crises only have a statistically significant effect on the income of the poor in richer countries. © 2017 The Authors Journal of International Development Published by John Wiley & Sons Ltd. Keywords: financial development; financial crises; poverty reduction
JEL Classification: G01; O15; I3 Keywords: financial development; financial crises; poverty reduction
JEL Classification: G01; O15; I3 *Correspondence to: Johan Rewilak, Aston Business School, Aston University, Birmingham, B4 7ET, United
Kingdom.
E-mail: j.rewilak@aston.ac.uk © 2017 The Authors Journal of International Development Published by John Wiley & Sons Ltd.
This is an open access article under the terms of the Creative Commons Attribution License, which permits use,
distribution and reproduction in any medium, provided the original work is properly cited. 1
INTRODUCTION Laeven and Valencia (2013) outline three different types of crises in their paper;
however, other types of financial crises exist, such as asset bubbles and inflation crises.1
We focus on banking, currency and debt crises, all which may have their own independent
effects on the poor. However, these different types of crises may occur simultaneously, or
one crisis may bring about the onset of another. Financial development is shown to be poverty reducing, where studies find that financial
deepening and increasing financial access to the poor may reduce poverty, (Burgess &
Pande, 2005; Beck, Demirgüc-Kunt, & Levine, 2007). Despite these findings, as financial
development increases, there is often an unintended consequence occurring simultaneously
—financial instability, (Akhter & Daly, 2009; Guillaumont Jeanneney & Kpodar, 2011). Financial instability may be harmful to society as it may disrupt payment systems and
harm long term growth. Moreover, it is usually the poor that are more vulnerable to
disruptions in the financial system compared to the rich. When the financial system suffers
a negative shock, banks begin to hoard liquidity resulting in a credit crunch.2 This may
damage society both in the short and long term. As banks increase their cost of credit to
recoup lost interest income (and increase profits), short term borrowing costs may rise,
but additionally, current reductions in credit reduce long run investment, harming long
run growth. In severe cases, when confidence in the banking sector completely evaporates,
bank runs and capital flight may occur that require immediate resolution. These may
include strict capital controls and short term bank closures until policy restores confidence
in the banking sector.3 These extreme measures are not only disruptive to the overall
functioning of the economy, but may severely harm the poor who may not have access
to other assets unlike the rich to help them during such times. A currency crisis is defined as a substantial devaluation or depreciation in the nominal
value of a currency, and this may have severe consequences for the poor. In particular, a
weakened currency makes imported goods more expensive. If imported goods have
inelastic demand, (food, energy and other necessities) the large surge in inflation may be
harmful to the economy. As the poor usually spend a far greater proportion of their overall
income on such necessities, they may be hardest hit. 1See (Reinhart & Rogoff, 2009) for the various types of crises and their definitions.
2With under capitalised banks, low profitability and uncertainty about credit quality, banks may hoard liquidity by
restricting lending until market conditions become clearer and the shock passes.
3During the Cypriot crisis of 2013, banks were closed for 2 weeks and a monthly cap was put in place on transfers
to foreign banks. 1
INTRODUCTION Whilst it is acknowledged that financial crises are harmful to the well-being of a country’s
citizens, little research has been carried out to quantify these effects. Notably, the effects of
financial crises may differ across the income distribution; and this study examines their
impact on the poor, who usually have limited resources at their disposal to insulate from
such negative shocks. We investigate episodes of banking, currency and debt crises using a new database from
Laeven and Valencia (2013) and measure the well-being of the poor as the income of the
lowest quintile in a population. This permits us to study crises in developed, as well as
under-developed nations, as financial crises are not just limited to developing countries. In our specifications, using a System GMM estimator with a lagged dependent variable,
we show that currency crises reduce the income of the poor by 14.9%, followed by 10.6%
for banking crises. Only in richer countries do we find that debt crises influence the income
of the poor. In addition, including a lagged dependent variable in the specification is © 2017 The Authors Journal of International Development Published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution License, which permits use,
distribution and reproduction in any medium, provided the original work is properly cited. J. Rewilak important, as numerous papers that study the effect of average growth on the income of the
poor (Dollar & Kraay, 2002; Rewilak, 2013) omit this variable. This may lead to the model
being misspecified. Nevertheless, the results confirm that whilst the short run coefficient
falls in size, in the long run, increases in average growth may be good for the poor, adding
a further valuable contribution to the literature. Financial crises are associated with periods of macroeconomic instability that disrupt the
normal functioning of the economy. Furthermore, during severe economic downturns, as
output falls below its natural rate, unemployment increases, and usually the first workers
to lose their jobs, are the low skilled poor. The financial crisis of 2007 began with problems
in the banking sector, but as public debt increased to bail out financial institutions, debt
crises emerged with further negative repercussions for the poor. In Greece, a prolonged
period of austerity reduced the state’s ability to create adequate safety nets for those in
need, further depressing their welfare. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. 1
INTRODUCTION In addition, as the poorest in society
are often in low paid jobs with little bargaining power, or are on fixed incomes such as J. Int. Dev. (2017)
DOI: 10.1002/jid J. Int. Dev. (2017)
DOI: 10.1002/jid The Impact of Financial Crises on the Poor pensions, inflation will reduce their purchasing power. In poor countries where the state is
a major employer, during a currency crisis, if the state is servicing foreign denominated
debt, these effects may be amplified as the government seeks cost saving measures. A
further way that the citizens may be harmed by a currency crisis is when the government
attempts to defend its currency. By raising domestic interest rates, policymakers may
prevent capital outflows; however, this comes at a cost. As interest rates increase to protect
the currency, local citizens now face higher repayments on personal debt, which was a
common feature during the East Asian crisis in 1997. Debt crises may be both internal and external, but their effects on domestic citizens, in
particular the poor are similar. Repayment of public debt requires either a reduction in
government spending or a tax increase or a combination of both. If income tax increases
are progressive, they may not be fully felt by the poor, but increases in consumption tax
will affect all citizens.4 The poor may feel this tax increase more than the rich, as their
marginal propensity to consume would theoretically be greater than their richer
counterparts. The other method that a government has at its disposal to manage a debt
crisis is to implement austerity programmes and cut public spending. For example, a
government may freeze public sector wages and reduce pro-poor expenditure such as
healthcare and education. As affluent members of society may not use such public services
(they can afford private health insurance, send their children to fee-paying schools), the
consequences of lower public spending is yet another way the poor may be harmed due
to debt crises. Financial crises usually reduce average incomes, and Demetriades, Rousseau and
Rewilak (2017) find that banking crises are negatively related with economic growth. If
increases in average income growth are associated with like-for-like increases in the
income growth of the poor (Dollar & Kraay, 2002), the poor will be harmed with lower
incomes during crisis episodes. 4In the United Kingdom, ad valorem tax was increased to 20 per cent in January 2011 as the government looked to
boost tax revenues to cut its deficit. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. 2
EMPIRICAL STRATEGY Equation (1) shows the benchmark estimation technique, fixed effects (FE). In Equation (1)
and all subsequent equations, (i) subscripts for individual countries, (t) subscripts for
individual years, where (α) and (β) are parameters to be estimated. The term (αi) represents
a country specific intercept, and the dependent variable is denoted (Y). Our variable of
interest (C) denotes the different types of financial crises, and matrix (X) contains our
covariates. The random error term (ϵ) is denoted. As we anticipate that our explanatory variables will be correlated with the error term,
pooled ordinary least squares (OLS) will be biased and inconsistent, and therefore we
prefer the fixed effects estimator. Whilst a random effects estimator may account for the
panel structure of the data, it too may be inconsistent. A Hausman test may be used to
determine the consistency of random effects, but regardless of the result, as we believe
our covariates are correlated with the error term, we use the fixed effects estimator. Y i;t ¼ αi þ β1Ci;t þ β2X i;t þ ϵi;t
(1) (1) Y i;t ¼ αi þ β1Ci;t þ β2X i;t þ ϵi;t In Equation (1), we assume our variable of interest (C) to be exogenous for the
following reasons. First, financial crises occur in both developed and developing countries,
therefore income or poverty should have no influence on whether a crisis will occur or
not.5 Likewise, financial crises occur in both equal and unequal societies, thus we argue
that simultaneity bias is not an issue. Second, we treat financial crises as exogenous
because they are unpredictable. If crises were predictable, then both national governments
and multilateral agencies would do their utmost to prevent them from occurring. Hence, we
can treat crises as a random shock. In Equation (2), a lagged dependent variable is introduced to the empirical specification. As the dependent variable, the income of the poor, is partly determined by inequality
which changes slowly through time, we believe previous values of the dependent variable
may be strong predictors of its current value. Leaving this unaddressed may bias the
estimates, but using a fixed effects estimator with a lagged dependent variable may
introduce a different type of bias into the estimates (Nickell, 1981), which only disappears
as (T) tends to infinity. However, the System GMM estimator proposed by (Blundell &
Bond, 1998) overcomes this problem. 5Banking crises have occurred in developing countries such as Venezuela in 1994 and similarly the global
financial crisis in 2007 hit a plethora of developed nations. Currency crises have hit developing countries/
emerging markets such East Asia 1997 and also developed nations, for example, Italy 1993. Finally, developing
nations such as Mexico in 1993 have had debt crises and recently developed nations such as the PIIGS (Portugal,
Ireland, Iceland, Greece and Spain) have found themselves on the cusp of debt crises. 1
INTRODUCTION On the other hand, Baldacci, de Mello and Inchauste
(2002) propose that as the poorest in society do not own property, or other tangible assets
that may lose a significant amount of their nominal value during a crisis, they may not
ultimately feel the burden of financial crises. Nevertheless, the authors do suggest that
when formal sector workers lose their jobs due to a crisis, they may enter the informal
sector. If those entering the informal sector are better educated, more productive and
efficient, then they may drive their poorer competitors out of the market. If this occurs,
once again, it is the poorest in society who lose out. There are several studies that examine the impact of financial crises on the poor’s
welfare. Gerry, Mickiewicz and Nikoloski (2014) show that currency crises may increase
mortality rates, as when the poor suffer negative income shocks associated with crises,
combined with increases in food prices, their nutritional levels fall and their health levels
deteriorate. Additionally, Habib, Narayan, Olivieri and Sanchez (2010) find that due to
the 2007 financial crisis, poverty reduction slowed in the Philippines and actually
increased in Mexico. These results compliment Chen and Ravallion (2009) who show that
at the aggregate level, the 2007 crisis will add a further 53 million people to the number
living below $1.25 a day globally, and that whilst aggregate poverty rates are still expected
to fall over time, they will do so at a slower rate. This paper adds to the literature by quantifying the effect of three different crises on the
income of the poorest quintile. Rather than focusing on just one specific crisis episode, it 4In the United Kingdom, ad valorem tax was increased to 20 per cent in January 2011 as the government looked to
boost tax revenues to cut its deficit. J. Int. Dev. (2017)
DOI: 10.1002/jid J. Int. Dev. (2017)
DOI: 10.1002/jid J. Rewilak J. Rewilak differentiates itself from the literature by focusing on all past banking, currency and debt
crises across multiple countries from 1973 to 2011. The rest of this paper is organised as
follows: Section 2 introduces the empirical strategy used in this paper, outlining the
methods and data. Section 3 presents the results and Section 4 concludes. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. 2
EMPIRICAL STRATEGY This estimator may also alleviate any remaining
endogeneity concerns by using instrumental variables. In addition, this estimator still
controls for country specific effects, but also permits the estimation of time invariant
variables, something the fixed effects estimator cannot do. J. Int. Dev. (2017)
DOI: 10.1002/jid J. Int. Dev. (2017)
DOI: 10.1002/jid The Impact of Financial Crises on the Poor Table 1. Summary statistics
Variable
Mean
Standard deviation
Minimum
Maximum
Income of the poor
11.10
1.14
7.94
13.44
GDP per capita
9.35
0.86
7.26
11.42
Private credit
3.64
0.89
1.68
5.74
Trade openness
91.35
44.11
15.64
348.39
Government spending
16.46
4.60
5.63
28.06
Inflation rate
3.01
1.14
11.99
1.41
Bank crisis
0.19
0.40
0.00
1.00
Currency crisis
0.03
0.16
0.00
1.00
Debt crisis
0.02
0.12
0.00
1.00
The following variables are entered as their natural logarithms as presented previously; income of the poor, GDP
per capita, and private credit. The inflation rate is first transformed so all values are positive and then logged. Trade openness and government spending are ratios to GDP and the crises variables are all dummy variables
taking a value equal to 1 if a crisis occurred and 0 otherwise. Table 1. Summary statistics The following variables are entered as their natural logarithms as presented previously; income of the poor, GDP
per capita, and private credit. The inflation rate is first transformed so all values are positive and then logged. Trade openness and government spending are ratios to GDP and the crises variables are all dummy variables
taking a value equal to 1 if a crisis occurred and 0 otherwise. Y i;t ¼ αi þ γY i;t1 þ β1Ci;t þ β2X i;t þ ϵi;t
(2)
ΔY i;t ¼ γΔY i;t1 þ β1ΔCi;t þ β2ΔX i;t þ Δϵi;t
(3) (2) (3) The System GMM estimator is shown in Equations (2) and (3). This estimator tests the
relationship between financial crises and the income of the poor as a system; and for it to
be valid, it requires that several diagnostic tests are satisfied. First, the estimator is designed
for a large number of cross sections and a smaller time dimension. In this paper, the
number of cross sections is 61 and the average number of time periods equals 9 satisfying
both conditions. The estimator requires the presence of first order serial correlation;
however, it requires no second order serial correlation. 6For further information regarding this estimator, we direct the reader to Baltagi (2008). © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. 2
EMPIRICAL STRATEGY In addition, the estimator uses
internal instruments to identify the parameters. In the level equation, the internal
instruments used are lagged differences of the endogenous variables and in the differenced
equation; the lagged levels are used as instruments. The System GMM estimator also
allows for external instruments to be used in the specification, in addition to any internal
instruments. In a recent paper examining financial crises and mortality, Gerry et al. (2014) include regional dummies as external instruments and this paper follows this
approach. The validity of the instruments may be tested using the Hansen test, where a
non-rejection of the null hypothesis indicates instrument validity. A further aspect of this
estimator is that it has a bias versus efficiency trade off. Whilst more instruments increase
the efficiency of the estimator, a large instrument count can overfit the endogenous
variables and bias the results. As there is no ideal instrument count, the literature
recommends that the number of instruments do not exceed the total number of cross
sections.6 We empirically address our research question using annual data for 61 countries from
1973 to 2011. The summary statistics are presented in Table 1. Our crisis data comes from
Laeven and Valencia (2013), and a value of one is assigned if a crisis has occurred in a
country in a given year and zero otherwise. The authors categorise crises into three
distinctive types: banking crises, currency crises and debt crises. The remaining variables
used in this study are available from the World Development Indicators website via the J. Int. Dev. (2017)
DOI: 10.1002/jid J. Int. Dev. (2017)
DOI: 10.1002/jid J. Rewilak World Bank. We select our control variables following Gerry et al. (2014) and these
variables are standard in the literature, appearing in many growth, poverty and other
macroeconomic studies. We control for GDP per capita and in its level form, the variable is highly skewed. As a
result, we take its natural logarithm which creates a more normal distribution. Moreover,
this allows us to compare our results with Dollar and Kraay (2002) who also apply a
logarithmic transform to this variable. Dollar and Kraay (2002) find that the coefficient
on this variable is equal to one, suggesting that any increase in average income is
associated with an analogous increase in the income of the poor. Furthermore, we control
for financial sector size using the natural logarithm of private credit. 2
EMPIRICAL STRATEGY We include it in the
specification as numerous studies show that providing finance to the poor may indeed
reduce poverty, (Akhter & Daly, 2009; Ang, 2010). Trade openness is measured as the
sum of exports and imports divided by GDP and government spending is measured using
final government expenditure divided by GDP. The expected sign on government
spending is ambiguous. Whilst pro-poor government spending may be beneficial for
poverty reduction, and redistribution programmes may reduce inequality, excess
government consumption has found to reduce economic growth in a plethora of studies. Moreover, if government spending is not pro-poor, with high amounts attached to
national defence, or spending on gentrification aimed at the middle and upper classes,
then this variable could enter the specifications negatively. To generate the inflation
variable we trim the data at the 1st and 99th percentile to remove influential
observations. We then add one to the variable so all values are strictly positive and take
its natural logarithm. We expect the inflation rate to enter the specifications negatively,
as the poor are usually on fixed incomes and do not have access to financial
instruments that overcome the negative effects of inflation. As a result, the poor’s
purchasing power would fall as would their real income. In extreme cases, the poor
may spend their time on cash management, as opposed to focusing on productive
activity that may help them generate income. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. 3
RESULTS Tables 2–4 present the benchmark findings. Each table examines the effect of a different
type of crisis on the income of the poor. Columns 1–3 present the fixed effects estimates,
whereas columns 4–6 introduce a lagged dependent variable into the specification and are
estimated using System GMM. The diagnostics of the System GMM estimates are
presented at the bottom of each table where p-values are reported for the AR(1) and
AR(2) test of serial correlation and for the Hansen test of instrument validity. Table 2 presents the results for banking crises where column 1 estimates a bivariate
regression, column 2 adds the covariates GDP per capita and private credit and column
3 adds the remaining control variables. In the bivariate regression, the banking crisis
variable is insignificant, but in the following two regressions, it enters with a negative
and significant sign. In columns 2 and 3, the only additional covariate that is significant
is GDP per capita. The results suggest that a 1% increase in average income transcends into
a 1.15–1.19% increase in the income of the poor. Columns 4–6 replicate the results from the previous three columns in Table 2. In the
bivariate regression, banking crises enter with a negative and significant sign, a result that
holds in the latter two columns. In the final and preferred specification, the results suggest J. Int. Dev. (2017)
DOI: 10.1002/jid J. Int. Dev. (2017)
DOI: 10.1002/jid J. Int. Dev. (2017)
DOI: 10.1002/jid The Impact of Financial Crises on the Poor Table 2. Effect of banking crises on the income of the poor
(1)
(2)
(3)
(4)
(5)
(6)
Crisis dummy
0.016 (0.22)
0.031a (1.77)
0.032a (1.79)
0.055c (3.18)
0.159c (2.99)
0.106c (3.63)
(Ln) GDP per capita
1.154c (16.05)
1.190c (14.82)
0.368a (1.80)
0.134c (3.84)
(Ln) Private credit
0.048 (1.35)
0.044 (1.18)
0.155 (1.63)
0.080c (3.05)
Trade openness
0.001 (1.01)
0.001b (2.52)
Government spending
0.003 (0.63)
0.017c (3.68)
(Ln) Inflation
0.018 (1.53)
0.001 (0.10)
(Ln) Lag income of poor
0.991c (173.71)
0.903c (7.37)
0.996c (40.06)
Number of instruments
19
29
54
AR(1) p-value
0.00
0.00
0.00
AR(2) p-value
0.25
0.16
0.13
Hansen p-value
0.12
0.30
0.21
Cross sections
61
61
61
61
61
61
Observations
514
514
514
514
514
514
Technique
FE
FE
FE
SYS-GMM
SYS-GMM
SYS-GMM
Each column represents a different regression and all models estimated using robust standard errors. T-statistics are reported in parentheses. 3
RESULTS adenotes the 10% statistical significance level
bdenotes the 5% statistical significance level
cdenotes the 1% statistical significance level
AR(1) is the test for first order serial correlation and AR(2) is the test for second order serial correlation. The Hansen test examines the validity of the instruments, where a non-
rejection indicates valid instruments. We use regional dummy variables as instruments in addition to internal instruments for regressions in columns 4–6. Authors Journal of International Development
John Wiley & Sons Ltd. J. D © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. J. Int. Dev. (2017)
DOI: 10.1002/jid J. Rewilak Table 3. Effect of currency crises on the income of the poor
(1)
(2)
(3)
(4)
(5)
(6)
Crisis dummy
0.332c (4.38)
0.058a (1.87)
0.058a (1.72)
0.137b (2.30)
0.088a (1.84)
0.149c (3.17)
(Ln) GDP per capita
1.110c (19.36)
1.185c (14.47)
0.280b (2.22)
0.126b (2.37)
Trade openness
0.001 (0.93)
0.002 (1.08)
0.003c (4.98)
0.001 (1.62)
(Ln) Private credit
0.045 (1.20)
0.091c (2.93)
Government spending
0.004 (0.83)
0.017c (3.65)
(Ln) Inflation
0.019 (1.52)
0.007 (0.78)
(Ln) Lag income of poor
0.982c (205.33)
0.590c (5.34)
0.994c (25.08)
Number of instruments
19
29
54
AR(1) p-value
0.00
0.00
0.00
AR(2) p-value
0.25
0.12
0.16
Hansen p-value
0.10
0.55
0.24
Cross sections
61
61
61
61
61
61
Observations
514
514
514
514
514
514
Technique
FE
FE
FE
SYS-GMM
SYS-GMM
SYS-GMM
Each column represents a different regression and all models estimated using robust standard errors. T-statistics are reported in parentheses. adenotes the 10% statistical significance level
bdenotes the 5% statistical significance level
cdenotes the 1% statistical significance level
AR(1) is the test for first order serial correlation and AR(2) is the test for second order serial correlation. The Hansen test examines the validity of the instruments, where a non-
rejection indicates valid instruments. We use regional dummy variables as instruments in addition to internal instruments for regressions in columns 4–6. wilak
e Authors Journal of International Development
J. Int J. Int. Dev. (2017)
DOI: 10.1002/jid © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. The Impact of Financial Crises on the Poor Table 4. 3
RESULTS Table 3 presents the results when the variable of interest is a currency crisis. In the
bivariate regression, the currency crisis dummy is negative and significant as anticipated
and this result holds across columns 2 and3. Similar to Table 2, the only covariate in
columns 2 and 3 that enters significantly is average income with an elasticity ranging from
1.10–1.19. In columns 4–6, the currency crisis dummy is negative and significant, and the results
suggest that a currency crisis may reduce the income of the poor between 9% and 15%,
where the highest magnitude is in column 6. In column 5, both controls average income
and trade openness are positive and significant, as is the lagged dependent variable. However, once including the remaining covariates in the final column, trade openness
becomes insignificant. The diagnostics for the models in the final three columns of
Table 3 are well satisfied as there is no presence of second order serial correlation and
the Hansen test shows the instruments appear to be valid. The instrument set is the same
as in Table 2 and does not exceed the number of cross sections. Table 4 examines the impact of debt crises on the income of the poor. The bivariate
regression in column 1 suggests that a debt crisis may reduce the poor’s income by
24%. However, once average income and government spending enter the regression, the
coefficient becomes insignificant. In column 3, when a full set of controls are used, the
only variable that is significant is average income. The results suggest that a 1% in average
incomes may increase the income of the poor by 1.19%, similar to the corresponding
regressions in Tables 2 and 3. Columns 4–6 present the results using the System GMM estimator. As in the previous
Tables, the lagged dependent variable is positive and highly significant across all three
columns. Whilst entering with its expected negative sign, the debt crisis variable is
statistically insignificant in columns 4–6. In the preferred regression in column 6, the
results suggest that increases in average income are good for the poor. In particular, a
1% increase in average income is associated with a 0.13% increase in the income of the
poor when examining the short run marginal effect. The Hansen test shows that the
instruments appear to be valid in all three specifications, with p-values ranging from
0.11–0.37. 3
RESULTS Effect of debt crises on the income of the poor
(1)
(2)
(3)
(4)
(5)
(6)
Crisis dummy
0.243b
(2.53)
0.039 (1.07)
0.019 (0.49)
0.099 (0.90)
0.080 (1.05)
0.097 (1.24)
(Ln) GDP per capita
1.099c (19.01)
1.191c (14.42)
0.233 (1.20)
0.128c (2.97)
Government spending
0.005 (0.87)
0.004 (0.75)
0.027c (2.73)
0.018c (5.13)
(Ln) Private credit
0.048 (1.26)
0.088c (3.66)
Trade openness
0.001 (1.01)
0.001c (2.82)
(Ln) Inflation
0.017 (1.44)
0.001 (0.06)
(Ln) Lag income of poor
0.983c (196.91)
0.757c (3.92)
0.988c (31.36)
Number of instruments
19
29
54
AR(1) p-value
0.00
0.01
0.00
AR(2) p-value
0.25
0.13
0.15
Hansen p-value
0.11
0.37
0.28
Cross sections
61
61
61
61
61
61
Observations
514
514
514
514
514
514
Technique
FE
FE
FE
SYS-GMM
SYS-GMM
SYS-GMM
Each column represents a different regression and all models estimated using robust standard errors. T-statistics are reported in parentheses. adenotes the 10% statistical significance level
bdenotes the 5% statistical significance level
cdenotes the 1% statistical significance level
AR(1) is the test for first order serial correlation and AR(2) is the test for second order serial correlation. The Hansen test examines the validity of the instruments, where a non-
rejection indicates valid instruments. We use regional dummy variables as instruments in addition to internal instruments for regressions in columns 4–6. Authors Journal of International Development
John Wiley & Sons Ltd. J. In
DO © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. J. Int. Dev. (2017)
DOI: 10.1002/jid J. Rewilak that a banking crisis may reduce the income of the poor by 10.6%.7 When introducing a
lagged dependent variable into the specification, the short run elasticity of average income
falls to 0.37% in column 5 and 0.13% in column 6. Private credit, whilst insignificant in
column 5 is negative and significant in the final column, whilst trade openness and
government spending are statistically significant in column 6. The diagnostics are well
satisfied in columns 4–6, where in all three regressions, there is evidence of first order
serial correlation and the null hypothesis of second order serial correlation is rejected
(p-values range from 0.13–0.25). The p-value of the Hansen test exceeds the traditional
level in all three regressions indicating that the instruments appear to be valid. Furthermore, the number of instruments does not exceed the number of cross sections as
proposed by Baltagi (2008). 7Column 6 is the preferred specification because it contains a full set of controls, including the lagged dependent
variable, which is shown to be very important based on its high statistical significance. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. 3
RESULTS In all three columns, the diagnostics show that there is no presence of second
order serial correlation. Furthermore, the number of cross sections exceeds the instrument
count as recommended in the literature. Table 5 presents the first set of robustness tests. Until now, each crisis was entered on
its own in the regressions; however, the different types of financial crises studied in this J. Int. Dev. (2017)
DOI: 10.1002/jid J. Int. Dev. (2017)
DOI: 10.1002/jid The Impact of Financial Crises on the Poor Table 5. Effect of all three crises on the income of the poor
(1)
(2)
(3)
Banking crisis dummy
0.040b (2.14)
0.091c (2.86)
0.100c (3.87)
Currency crisis dummy
0.093a (1.87)
0.139b (2.35)
0.140c (2.84)
Debt crisis dummy
0.048 (0.56)
0.012 (0.19)
0.025 (0.37)
(Ln) GDP per capita
0.130c (4.51)
0.136c (4.44)
(Ln) Private credit
0.070c (3.18)
0.070c (3.05)
Trade openness
0.001 (1.62)
0.001b (2.56)
Government spending
0.014c (3.64)
0.015c (3.67)
(Ln) Inflation
0.009 (0.91)
0.014 (1.45)
(Ln) Lag income of poor
0.990c (193.99)
0.987c (48.70)
0.989c (44.99)
Number of instruments
33
68
59
AR(1) p-value
0.00
0.00
0.00
AR(2) p-value
0.20
0.13
0.12
Hansen p-value
0.18
0.66
0.37
Cross sections
61
61
61
Observations
514
514
514
Technique
SYS-GMM
SYS-GMM
SYS-GMM
Each column represents a different regression and all models estimated using robust standard errors. T-statistics
are reported in parentheses. adenotes the 10% statistical significance level
bdenotes the 5% statistical significance level
cdenotes the 1% statistical significance level
AR(1) is the test for first order serial correlation and AR(2) is the test for second order serial correlation. The
Hansen test examines the validity of the instruments, where a non-rejection indicates valid instruments. We use
regional dummy variables as instruments in addition to internal instruments for all the regressions. Table 5. Effect of all three crises on the income of the poor paper may occur simultaneously. Table 5 contains three columns. In column 1, only the
three different types of crises are entered into the regression along with the lagged
dependent variable. Column 2 then adds the remaining covariates into the regression
specification. Finally, column 3 re-estimates the regression in column 2 using a smaller
instrument set. In column 1, it is evident that both banking and currency crises are negative and
significant, whereas debt crises, although negative in sign, are insignificantly related to
the income of the poor. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd.
J. Int. Dev. (2017)
DOI: 10.1002/jid 3
RESULTS Examining the results from column 2, only trade openness, debt
crises and the inflation rate are insignificant, where both banking and currency crises retain
their negative signs. However, by including an extra two variables in the specification, the
number of instruments exceeds the number of cross sections. According to Baltagi (2008)
having the number of instruments exceed, the number of cross sections may overfit the
endogenous regressors and bias the results. Therefore in column 3, the number of
instruments is reduced and the regression in column 2 is re-estimated. The results show
that the coefficients in column 3 are very similar to those in column 2 providing confidence
in the results. In column 3, the results suggest that banking crises may reduce the income of
the poor by approximately 10% and currency crises reduce the income of the poor by 14%. The coefficients are similar in size to those in the previous tables, and it can be confidently
stated, that the impact of financial crises on the income of the poor was not over-estimated
in the previous tables. Table 6 examines whether the effects of financial crises differ depending on the level of
financial development, trade openness or government expenditure. Therefore, Table 6
includes interaction terms between the different types of crises and these variables. J. Int. Dev. (2017)
DOI: 10.1002/jid J. Rewilak Table 6. Financial crises and the income of the poor: Interaction terms
(1)
(2)
(3)
Crisis dummy
0.158c (2.95)
0.148c (2.60)
0.097 (1.20)
Crisis × private credit
0.054 (1.32)
Crisis × trade openness
0.005 (0.89)
Crisis × government spending
0.002 (0.13)
(Ln) GDP per capita
0.136c (3.53)
0.137c (3.96)
0.126c (3.30)
(Ln) Private credit
0.082c (3.47)
0.093c (3.58)
0.088c (3.63)
Trade openness
0.001b (2.30)
0.001c (3.00)
0.001c (3.18)
Government spending
0.016c (4.24)
0.017c (4.89)
0.018c (4.90)
(Ln) Inflation
0.005 (0.49)
0.003 (0.31)
0.000 (0.04)
(Ln) Lag income of poor
0.986c (34.37)
0.984c (48.36)
0.990c (36.45)
Number of instruments
54
54
54
AR(1) p-value
0.00
0.00
0.00
AR(2) p-value
0.17
0.16
0.16
Hansen p-value
0.25
0.21
0.19
Cross sections
61
61
61
Observations
514
514
514
Technique
SYS-GMM
SYS-GMM
SYS-GMM
Each column represents a different regression and all models estimated using robust standard errors. Columns 1
reports results for banking crises, column 2 for currency crises and columns 3 for debt crises. T-statistics are
reported in parentheses. 3
RESULTS adenotes the 10% statistical significance level
bdenotes the 5% statistical significance level
cdenotes the 1% statistical significance level
AR(1) is the test for first order serial correlation and AR(2) is the test for second order serial correlation. The
Hansen test examines the validity of the instruments, where a non-rejection indicates valid instruments. We use
regional dummy variables as instruments in addition to internal instruments for all the regressions. Table 6. Financial crises and the income of the poor: Interaction terms Each column represents a different regression and all models estimated using robust standard errors. Columns 1
reports results for banking crises, column 2 for currency crises and columns 3 for debt crises. T-statistics are
reported in parentheses. Each column represents a different regression and all models estimated using robust standard errors. Columns 1
reports results for banking crises, column 2 for currency crises and columns 3 for debt crises. T-statistics are
reported in parentheses. AR(1) is the test for first order serial correlation and AR(2) is the test for second order serial correlation. The
Hansen test examines the validity of the instruments, where a non-rejection indicates valid instruments. We use
regional dummy variables as instruments in addition to internal instruments for all the regressions. In column 1, the banking crisis dummy is interacted with private credit, as banking
crises may potentially harm the poor more in underdeveloped banking systems compared
to well-developed banking sectors. This is because well-developed banking sectors usually
have greater provisions in place that may safeguard customer deposits, or even be better
equipped to keep lending through turbulent times. The results show that whilst the crisis
dummy retains its negative sign and statistical significance, the interaction term between
banking crises and credit is insignificant. Nevertheless, the interaction term is positive,
which implies that larger financial sectors may reduce the negative impact of a banking
crisis on the income of the poor. All the remaining covariates are statistically significant,
with signs consistent from the previous estimations, with the exception of the inflation rate. In column 2, the currency crisis dummy is interacted with trade openness, as the impact
of a crisis on the income of the poor, may depend on the openness of an economy. As in
column 1, the crisis dummy is negative and significant with a magnitude similar to previous
estimates, although the interaction term is positive and insignificant. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd.
J. Int. Dev. (2017)
DOI: 10.1002/jid 3
RESULTS The positive
coefficient on the interaction term suggests that similar to banking crises, the more open
an economy is, the less damaging a currency crisis may be to the income of the poor. A
crisis that devalues the currency may be a blessing if it makes a country more competitive
internationally. In export-oriented labour intensive industries, following a depreciation in
the currency, product demand may increase. The knock on effect here may increase
employment or wages in this sector. As it is usually the poor who work in such sectors, then
they may actually benefit from a currency devaluation. The control variables in column 2
are very similar to those in column 1, where only the inflation rate enters insignificantly. J. Int. Dev. (2017)
DOI: 10.1002/jid J. Int. Dev. (2017)
DOI: 10.1002/jid The Impact of Financial Crises on the Poor The final column of Table 6 interacts debt crises with government spending. The impact
of a debt crisis on the poor may be greater in countries with higher government spending as
opposed to countries with lower spending. For example, if government expenditure is
close to zero, then further reductions in spending due to a crisis would be negligible and
may not impact society compared with larger reductions in government spending. The
results suggest that the coefficient on debt crises is negative and insignificant, as is the
interaction term. Furthermore, the remaining covariates are consistent with the previous
two regressions. The diagnostics in Table 6 are all well satisfied with Hansen p-values
all exceeding 0.19. There is no presence of second order serial correlation with AR(2)
p-values between 0.16–0.17 and in all three specifications the number of cross sections
exceeds the instrument count. Table 7 examines whether the effects of financial crises on the income of the poor is
homogeneous across the large number of countries. As the System GMM estimator
requires that the number of cross sections is large, splitting the sample into multiple
income groups or geographic regions would violate this assumption. As a result, the
sample was split to investigate the effects of the three types of financial crises on the
poorest 30 countries in the sample and richest 31 countries in the sample. Columns 1
and 2 present the results for banking crises, columns 3 and 4 for currency crises and the
final two columns examine the effects of debt crises. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. 3
RESULTS Columns 1 and 2 show that banking crises are harmful to the poor’s income, both in
poorer and richer countries in the sample. The magnitude for poorer countries is
approximately 9%, whereas for richer countries it is close to 13%. This is an interesting
finding as it may capture the fact that in some of the poorest nations in the world, many
citizens do not have access to formal financial services. Therefore, if a banking crisis
occurs, the poor who are excluded from finance may not face as serious consequences as
those who are included in the financial system, possibly explaining the difference in the
magnitude of the coefficients. In the middle two columns, the results are very similar to columns 1 and 2 where a
currency crisis will reduce the income of the poor in both poorer and richer countries. However, the impact on poorer countries is lower than that of richer countries, an
interesting finding which may potentially be explained for the following reasons. First,
as it is usually richer countries who engage in international capital markets, and as
currency devaluations increase foreign denominated debt, only participants in such
markets will be harmed via this channel. Additionally, imported inflation, another
consequence of currency devaluations, may be better overcome by low income households
in poorer countries compared with low income households in richer countries. In
developing economies, with a greater reliance on agriculture and the experience to trade
using a barter economy, the poor may be able to avoid dealing with cash whose value is
frequently changing and falling. On the other hand, in OECD nations, due to the
industrialised nature of their economies, the poor may not have such options readily
available to them, as most of the individuals in the lowest income quintile would be
working in secondary or tertiary sectors and would be paid in cash, rather than in goods
and services. Finally, columns 5 and 6 show the impact of debt crises on the income of the poor is
negative and significant only in the richer sub-sample of countries. In previous estimates
debt crises were always insignificant, thus, splitting the sample unearths a further
important finding. However, as richer economies may be in a better position to target the
poor with the governmental expenditure, this result is not overly surprising. J. Int. Dev. (2017)
DOI: 10.1002/jid J. Int. Dev. (2017)
DOI: 10.1002/jid J. 3
RESULTS The Impact of Financial Crises on the Poor The Impact of Financial Crises on the Poor The diagnostics are generally satisfied in Table 7. However, because the sample was
split and the number of cross sections reduced, in order to avoid overfitting the endogenous
regressors, the instrument set had to be reduced. This was performed by treating only
average income, the financial crises and the lagged dependent variable as endogenous. Therefore, it is important to treat the findings in Table 7 with caution. Overall, the results show that different types of crises may have different impacts on the
income of the poor. In the preferred specifications, both bank and currency crises
negatively influence the income of the poor, but debt crises only have a negative and
significant effect for the richest countries in the sample. Examining the covariates, private
credit is found to be harmful in raising the income of the poor. This is somewhat surprising
as finance has been shown to be beneficial in poverty reduction. However, studies show
that there are certain conditions for finance to be pro-growth and pro-poor. Rewilak
(2013) shows that whilst increasing financial sector development may reduce poverty in
certain worldwide regions, in others it may detrimental. Rioja and Valev (2004) show that
finance is most beneficial for growth in middle income countries, a result that may extend
to poverty reduction, potentially explaining this finding.8 This finding is well demonstrated
in Table 7 as finance is only negative and significant in the subsample of poorer countries. Government spending also enters negatively in the specifications but this is not overly
surprising. Increases in government spending such as redistribution programmes may
increase the incomes of the poor, but if this spending fails to reach the target population,
or if the government engages in wasteful expenditure, for example beautifying government
buildings, it may be harmful to the income of the poor if their taxes are paying for such
projects. 8Further evidence, by Nikoloski (2013), shows that a financial Kuznets curve may exist, where increasing
financial sector development may initially lead to higher rates of inequality before eventually falling. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. 3
RESULTS Rewilak Table 7. Financial crises and the income of the poor: Splitting the sample
(1)
Low income
(2)
High income
(3)
Low income
(4)
High income
(5)
Low income
(6)
High income
Crisis dummy
0.094b (2.53)
0.129c (3.83)
0.137a (1.92)
0.195c (2.64)
0.064 (0.58)
0.171c (3.20)
(Ln) GDP per capita
0.117a (1.90)
0.084c (2.86)
0.106b (2.10)
0.084c (3.10)
0.121b (2.11)
0.069c (3.03)
(Ln) Private credit
0.061b (2.55)
0.011 (0.75)
0.059c (3.10)
0.021 (1.19)
0.067b (2.29)
0.014 (1.37)
Trade openness
0.000 (0.28)
0.000 (1.62)
0.000 (0.39)
0.000a (1.84)
0.000 (0.51)
0.000c (2.69)
Government spending
0.004 (1.05)
0.004 (1.01)
0.004 (0.93)
0.004a (1.80)
0.003 (0.69)
0.004 (1.29)
(Ln) Inflation
0.009 (1.23)
0.014b (2.53)
0.012 (1.45)
0.003 (0.39)
0.005 (0.67)
0.005 (0.67)
(Ln) Lag income of poor
0.970c (25.97)
0.954c (59.84)
0.965c (27.43)
0.937c (49.45)
0.954c (25.89)
0.945c (63.71)
Number of instruments
29
30
29
30
29
30
AR(1) p-value
0.00
0.06
0.00
0.10
0.00
0.08
AR(2) p-value
0.39
0.20
0.44
0.23
0.41
0.28
Hansen p-value
0.20
0.24
0.21
0.18
0.16
0.29
Cross sections
30
31
30
31
30
31
Observations
290
224
290
224
290
224
Technique
SYS-GMM
SYS-GMM
SYS-GMM
SYS-GMM
SYS-GMM
SYS-GMM
Each column represents a different regression and all models estimated using robust standard errors. Columns 1 and 2 report results for banking crises, columns 3 and 4 for currency
crises and columns 5 and 6 for debt crises. T-statistics are reported in parentheses. adenotes the 10% statistical significance level
bdenotes the 5% statistical significance level
cdenotes the 1% statistical significance level
AR(1) is the test for first order serial correlation and AR(2) is the test for second order serial correlation. The Hansen test examines the validity of the instruments, where a non-
rejection indicates valid instruments. In this Table, only financial crises, average income and the lagged dependent are treated as endogenous to ensure the instrument count does
not exceed the number of cross sections. In addition to the internal instruments, regional dummies are included as additional instruments in all the regressions. e Authors Journal of International Development
by John Wiley & Sons Ltd. J. Int. D
DOI: 1 J. Int. Dev. (2017)
DOI: 10.1002/jid © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. J. Int. Dev. (2017)
DOI: 10.1002/jid © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. 4
CONCLUSION This study investigates whether financial crises have a negative impact on the income of
the lowest quintile. This permits us to examine the role of financial crises on the poor in
both developed and developing countries. Three different types of crises are examined in
this analysis, which include banking, currency and debt crises. The results show that a banking crisis may harm the income of the poor by 10.6%. When
the banking system comes under distress, methods of payment may shut down. If the poor
are relying on financial institutions for remitted payments, they may suffer from temporary
income shortfalls, or may have to use more expensive methods to receive their funds,
which in turn will reduce the total payment received. Bank closures may result in a
shortage of liquidity and as a method to cope, the poor may sell off their non-monetary
assets, possibly at a discount, jeopardising their wealth. Finally, if a financial institution
becomes insolvent and is not backed by a government guarantee, depositors may lose their
money. Currency crises are found to have the most detrimental impact on the poor, where a
currency crisis may reduce the income of the lowest quintile by 14.9%. Whilst, a currency
depreciation may result in higher remitted payments in local currency from abroad, the
negative consequences of a depreciation may more than offset this benefit. Imported
inflation will make the cost of living more expensive, especially if imported goods have J. Int. Dev. (2017)
DOI: 10.1002/jid J. Int. Dev. (2017)
DOI: 10.1002/jid J. Rewilak a low elasticity of demand. Even if the poor consume local goods, if intermediate goods are
imported, then producers may pass these costs onto the consumer and local prices will still
rise. If a country has been fighting devaluation, interest rate increases to defend the
currency may hurt the poor if they have obtained credit, as the poor will face higher loan
repayments. Moreover, with higher interest rates, even if the poor are not borrowers, they
may be subject to higher rental fees as landowners look to pass on their higher costs. Debt crises are only shown to have a statistically significant effect on the income of the
poor in richer countries. Whilst debt crisis may result in higher taxes and lower social
spending, these effects may not be wholly felt by the poor in less affluent countries. ACKNOWLEDGEMENTS I would like to extend my thanks to delegates at the 49th MMF conference, the Journal of
International Development including the editorial team, in addition to two anonymous
referees. Their comments and suggestions have led to substantial improvements to this
paper. Johan Rewilak acknowledges support of ESRC-DFID grant number ES/J009067/1. 4
CONCLUSION If
the poor do not work in the formal sector, they will not be subject to income tax and are
unaffected by tax increases as a government seeks to generate revenue. Indeed, if the tax
system is progressive and the tax increases are felt at the higher end of the income
distribution, once more the poor may be unaffected. A government reducing social
spending may be harmful to the poor; however, this depends on how well a government
targets its spending. If a government is incorrectly targeting the poor, then they should feel
no effects to cuts in spending. Moreover, spending cuts may end up harming the middle
classes (those higher up in the income distribution), a question I leave unaddressed for
future research. In this study, we uncover a further valuable contribution to the academic literature. By
including a lagged dependent variable in the specifications, we still find evidence that
growth may be good for the poor. Nevertheless, as financial crises depress economic
growth, they may have additional consequences on the poor, and to what extent requires
further research, which may stimulate future policy debate. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd.
J. Int. Dev. (2017)
DOI: 10.1002/jid © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. REFERENCES Akhter S, Daly K. 2009. Finance and poverty: evidence from fixed effect vector decomposition. Emerging Markets Review 10: 191–206. Ang J. 2010. Finance and inequality: the case of India. Southern Economic Journal 76: 738–761. Baldacci C, de Mello L, Inchauste G. 2002. Financial crisis, poverty and income distribution. I
Working Paper, 02/04. Washington DC: IMF. Baltagi B. 2008. Econometric Analysis of Panel Data. John Wiley & Sons Ltd: Chichester. Beck T, Demirgüc-Kunt A, Levine R. 2007. Finance, inequality, and poverty: cross-country
evidence. Journal of Economic Growth 12: 21–47. Blundell R, Bond S. 1998. Initial conditions and moment restrictions in dynamic panel data models. Journal of Econometrics 87: 115–143. Burgess R, Pande R. 2005. Do rural banks matter? Evidence from the Indian social banking
experiment. American Economic Review 95: 780–795. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. J. Int. Dev. (2017)
DOI: 10.1002/jid J. Int. Dev. (2017)
DOI: 10.1002/jid J. Int. Dev. (2017)
DOI: 10.1002/jid The Impact of Financial Crises on the Poor Chen, S., Ravallion, M., 2009. The impact of the global financial crisis on the world’s poorest. World
Bank Development Research Group. Vox-EU. Demetriades P, Rousseau P, Rewilak, J. 2017. Finance, growth and fragility, Discussion Papers in
Economics 17/13, Department of Economics, University of Leicester. UK. Dollar D, Kraay A. 2002. Growth is good for the poor. Journal of Economic Growth 7: 195–225. Gerry C, Mickiewicz T, Nikoloski Z. 2014. Mortality and financial crises. Journal of International
Development 26: 939–948. Guillaumont Jeanneney S, Kpodar K. 2011. Financial development and poverty reduction: can ther Guillaumont Jeanneney S, Kpodar K. 2011. Financial development and poverty reduction: can there
be a benefit without a cost? Journal of Development Studies 47: 143–163. be a benefit without a cost? Journal of Development Studies 47: 143–163. Habib B, Narayan A, Olivieri S, Sanchez C. 2010. The impact of the financial crisis on poverty and
income distribution: insights from simulations in selected countries. World Bank Economic
Premise 7: 1–4. Laeven L, Valencia F. 2013. Systemic banking crises database: an update. IMF Working Paper
12/163. Washington DC: IMF. Nickell S. 1981. Biases in dynamic models with fixed effects. Econometrica 49: 1417–1426. Nikoloski Z. 2013. Financial sector development and inequality: is there a financial Kuznets curve? Journal of International Development 25: 897–911. Reinhart C, Rogoff K. 2009. This Time Is Different: Eight Centuries of Financial Folly. © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd. J. Int. Dev. (2017)
DOI: 10.1002/jid REFERENCES Princeton
University Press: Princeton, New Jersey. https://press.princeton.edu/titles/8973.html Rewilak J. 2013. Finance is good for the poor but it depends where you live. Journal of Banking &
Finance 37: 1451–1459. Rioja F, Valev N. 2004. Does one size fit all? A re-examination of the finance and growth
relationship. Journal of Development Economics 74: 429–447. J. Int. Dev. (2017)
DOI: 10.1002/jid © 2016 The Authors Journal of International Development
Published by John Wiley & Sons Ltd.
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ESTADO, POLÍTICAS EDUCACIONAIS E TECNOLOGIAS FRENTE ÀS DEMANDAS DO CAPITALISMO CONTEMPORÂNEO
|
Maria Sueli Corrêa dos Prazeres
|
Portuguese
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Spoken
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Práxis
Educacional
e-ISSN 2178-2679
Revista
ARTIGO
https://doi.org/10.22481/praxis.v15i32.5060 Práxis
Educacional
e-ISSN 2178-2679
Revista
ARTIGO
https://doi.org/10.22481/praxis.v15i32.5060 e-ISSN 2178-2679 ARTIGO
https://doi.org/10.22481/praxis.v15i32.5060 ESTADO, POLÍTICAS EDUCACIONALES Y TECNOLOGÍAS FRENTE A LAS
DEMANDAS DEL CAPITALISMO CONTEMPORÁNEO Maria Sueli Corrêa dos Prazeres
Universidade Federal do Pará – Brasil Maria Sueli Corrêa dos Prazeres
Universidade Federal do Pará – Brasil Ilda Gonçalves Batista
Universidade Federal do Pará – Brasil Ilda Gonçalves Batista
Universidade Federal do Pará – Brasil Resumo: O presente artigo objetiva analisar o Estado e sua articulação na formulação das políticas
educacionais voltadas a inserção das tecnologias digitais na educação. Faz-se um convite à reflexão,
no sentido de promover, a compreensão de como o capital age, junto ao Estado, no processo de
modelamento das políticas educacionais aos ideais econômicos e suas demandas a conjuntura
educacional em face às intencionalidades de reestruturação econômico-produtiva. Adota-se uma
análise crítica da tecnologia, considerada a partir do contexto histórico vivido e no conjunto da
realidade social no qual é projetada, com o intuito de problematizar sua relação direta com a produção
capitalista. Reflete-se sobre a trama de consolidação das TICs na educação demonstrando que a
política de inclusão digital apresenta-se como um projeto capitalista de ajuste do Estado brasileiro às
pressões políticas dos países hegemônicos para o qual as novas tecnologias atuam em benefício e para
promoção do capital, numa busca incessante por novas formas de se solidificar economicamente. Palavras-chave: Estado; Políticas Educacionais; Tecnologias de Informação e Comunicação. Abstract: The present article aims to analyze the State and its articulation in the formulation of
educational policies aimed at the insertion of digital technologies in education. It is an invitation to the
reflection, in the sense to promote, the understanding of how capital acts, with the State, in the process
of modeling educational policies to economic ideals and their demands to the educational situation in
the face of the intentionalities of economic-productive of restructuring. A critical analysis of
technology is adopted, considered started on the lived historical context and the social reality in which
it is projected, intent to problematize your direct relation with capitalist production. It reflects on the
plot of consolidation of ICT in education demonstrating that the politic of digital inclusion presents
itself as a capitalist project of adjustment of the Brazilian State to the political pressures of the
hegemonic countries for which the new technologies act for the benefit and for the promotion of the
capital, in an incessant search for new ways to solidify economically. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. ESTADO, POLÍTICAS EDUCACIONALES Y TECNOLOGÍAS FRENTE A LAS
DEMANDAS DEL CAPITALISMO CONTEMPORÁNEO 378 Práxis
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Keywords: State; Educational Policies; Information and Communication Technologies. Revista e-ISSN 2178-2679 Keywords: State; Educational Policies; Information and Communication Technologies. Resumen: El presente artículo busca analizar al Estado y su articulación con la formulación de las
políticas educacionales orientadas hacia la inserción de las tecnologías digitales en la educación. Se
llama a la reflexión, en el sentido de promover la comprensión, sobre como actúa el capital junto al
Estado en el proceso de ajuste de las políticas educacionales a los ideales económicos y sus demandas,
la coyuntura educacional frente a las intenciones de reestructuración económico-productiva. Se adopta
un análisis crítico de la tecnología, considerada a partir del contexto histórico vivido y en el conjunto
de la realidad social hacia la cual es proyectada, con la intención de problematizar su relación directa
con la producción capitalista. Se refleja sobre la trama de consolidación de las TICs en la educación
demostrando que la política de inclusión digital se presenta como un proyecto capitalista de ajuste del
Estado brasilero a las presiones políticas de los países hegemónicos, que se benefician con las nuevas
tecnologías, y para la promoción del capital, en una búsqueda incesante de nuevas formas de
consolidarse económicamente. bras clave: Estado; Políticas Educacionales; Tecnologías de Información y Comunicación. Palabras clave: Estado; Políticas Educacionales; Tecnologías de Información y Comunicación. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. Introdução Na atualidade, vivem-se intensas transformações frutos dos avanços científicos e
tecnológicos dos últimos anos que remodelam a sociedade em ritmo acelerado. Designada de
“Revolução Informacional ou da Informação” (CASTELLS, 1999), seria ela uma sequência
histórica das revoluções sucessivas ocorrentes a partir da aceleração do processo e da
veiculação da informação através da utilização da tecnologia. Para este autor uma das
principais características dessa revolução tecnológica é a aplicação dos conhecimentos e das
informações em um ciclo de realimentação cumulativo entre a inovação e o uso, no qual as
novas tecnologias de informação e comunicação não são simplesmente ferramentas a serem
aplicadas, mas sim processos a serem desenvolvidos. Este trabalho problematiza que as novas tecnologias surgem em benefício e para
promoção do capital, numa busca incessante por novas formas de se solidificar
economicamente. Nesse processo, o Estado e o capital agem ideologicamente em defesa de
sua manutenção e, assim, acabam por atribuir à educação e às tecnologias, condições
necessárias para criar uma base mais sólida de produção, onde impera a busca por melhor
qualificação para o mercado de trabalho com objetivo de gerar lucro e riqueza. Assim, faz-se um convite à reflexão, no sentido de promover, a compreensão de como
o capital age, junto ao Estado, no processo de modelamento das políticas educacionais aos
ideais econômicos em face às intencionalidades de reestruturação econômico-produtiva. 379 Práxis
Educacional
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e-ISSN 2178-2679
Revista
Parte-se da análise do contexto histórico de produção da sociedade capitalista e sua relação
com o Estado enquanto produtores de demandas tecnológicas a conjuntura educacional na
contemporaneidade Práxis
Educacional
Revista Revista e-ISSN 2178-2679 Parte-se da análise do contexto histórico de produção da sociedade capitalista e sua relação
com o Estado enquanto produtores de demandas tecnológicas a conjuntura educacional na
contemporaneidade. Parte-se da análise do contexto histórico de produção da sociedade capitalista e sua relação
com o Estado enquanto produtores de demandas tecnológicas a conjuntura educacional na
contemporaneidade. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019 Relação estado-capital no delineamento das políticas educacionais 380 Práxis
Educacional
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Revista Práxis
Educacional
Revista Revista e-ISSN 2178-2679 e-ISSN 2178-2679 É nesse sentido que o Estado adota, por meio de suas ações sociais (na educação,
saúde, etc.), papel inexorável nas práticas de manutenção do capital, constituindo-se como
requisito indispensável para sua produção e, no dizer de Mészáros (2011), um “microcosmo
sociometabólico do capital”, ou seja, uma unidade de reprodução socioeconômica, capaz de
fazer funcionar, por meio de sua estrutura de comando e controle político, as estruturas
reprodutivas do capital adaptando os diversos órgãos sociais que atuam sobre a regra deste
sistema. Ball (2004) adverte que: Cada vez mais, as políticas sociais e educacionais estão sendo articuladas e
legitimadas explicita, direta e, muitas vezes exclusivamente em função do
seu papel em aumentar a competitividade econômica por meio do
desenvolvimento das habilidades, capacidades e disposições exigidas pelas
novas formas econômicas da alta modernidade. (BALL, 2004, p. 1109). Cada vez mais, as políticas sociais e educacionais estão sendo articuladas e
legitimadas explicita, direta e, muitas vezes exclusivamente em função do
seu papel em aumentar a competitividade econômica por meio do
desenvolvimento das habilidades, capacidades e disposições exigidas pelas
novas formas econômicas da alta modernidade. (BALL, 2004, p. 1109). Cabe destacar que nesse processo o Estado age como meio articulador dos interesses
da economia capitalista enquanto poderoso instrumento de exploração política a favor dos
grandes interesses do capital e, de uma classe, com o objetivo de remover as barreiras que
dificultam seu desenvolvimento. Necessariamente, nesse cenário o Estado exerce uma dupla
exploração “enquanto explora como qualquer outro capitalista, e enquanto aparelho repressivo
e ideológico, efetiva uma exploração política a favor do capital no seu conjunto”
(FRIGOTTO, 2006, p. 102), inserindo políticas que vislumbrem “permanecer estritamente
dentro dos limites da perpetuação do domínio do capital como modo de reprodução social
metabólica” (MÉSZÁROS, 2008, p. 26). Para Ball (2004), na sociedade do capital as instituições do setor público estão sendo
“repensadas” como oportunidades de lucros e, cada vez mais, o mundo dos negócios enfoca
os serviços de educação como uma área em expansão, na qual lucros consideráveis devem ser
obtidos. De modo geral, para que isso se efetive é imprescindível que o Estado em sua postura
reguladora gere condições para o capital investir e proliferar. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. Relação estado-capital no delineamento das políticas educacionais O capital necessita de estruturas que viabilizem seu controle na sociedade, assim, a
formação do Estado moderno “é uma exigência absoluta para assegurar e proteger
permanentemente a produtividade do sistema” (MÉSZÁROS, 2011, p. 106), no qual suas
práticas políticas atuam paralelamente a este, para favorecimento da dinâmica mutante de sua
expansão e acumulação. Dessa forma, no sistema capitalista, o Estado é levado a “mediar os interesses
intercapitalistas e preservar o sistema como um todo” (FRIGOTTO, 2006, p. 100), para
cumprir seu movimento e com isso alavancar seu crescimento. Para Frigotto (2006), este se
assume como a própria forma do modo de produção capitalista enquanto gerenciador das suas
relações dentro do movimento de acumulação, concentração e centralização do capital. O Estado moderno constitui a “única estrutura corretiva compatível com os parâmetros
estruturais do capital como modo de controle sociometabólico” (MÉSZÁROS, 2011, p. 107),
protege legalmente a relação de forças estabelecidas, logo, sem essa estrutura jurídica não há
como o capital manter sua eficiência econômica. Assim, como parte integrante da própria
base material do capital, o Estado tem papel significativo na formação e consolidação de todas
as grandes estruturas reprodutivas da sociedade: O Estado moderno altamente burocratizado, com toda a complexidade de seu
maquinário legal e político, surge da absoluta necessidade material da ordem
metabólica do capital e depois, por sua vez – na forma de uma reciprocidade
dialética – torna-se uma precondição essencial para a subsequente
articulação de todo o conjunto. (MÉSZÁROS, 2011, p. 108). O Estado moderno altamente burocratizado, com toda a complexidade de seu
maquinário legal e político, surge da absoluta necessidade material da ordem
metabólica do capital e depois, por sua vez – na forma de uma reciprocidade
dialética – torna-se uma precondição essencial para a subsequente
articulação de todo o conjunto. (MÉSZÁROS, 2011, p. 108). Para Mészáros (2011), isso significa dizer que o Estado se afirma como pré-requisito
indispensável para o funcionamento permanente do sistema do capital nas suas mais ínfimas
relações, não impondo barreiras para sua expansão. Deste modo, o Estado assume papel
totalizador e essencial nesse modelo de sociedade, pois deve sempre ajustar suas funções
reguladoras em sintonia com a dinâmica do processo de reprodução socioeconômica,
completando politicamente e reforçando a dominação do capital. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. Relação estado-capital no delineamento das políticas educacionais Dessa forma, Barroso (2005) argumenta que a regulação realizada pelo Estado é vista
como uma função essencial para a manutenção do equilíbrio de qualquer sistema (físico ou
social) e está associada aos processos de retroação (positiva ou negativa). É ela que permite
ao sistema, através dos seus órgãos reguladores, identificar as perturbações, analisar e tratar as
informações relativas a um estado de desequilíbrio e transmitir um conjunto de ordens
coerentes a um ou vários dos seus órgãos executores. Assim, a regulação está associada ao
objetivo de consagrar à intervenção do Estado na condução das políticas públicas. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. 381 Práxis
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Revista Revista e-ISSN 2178-2679 O campo educacional, ao longo do processo histórico de desenvolvimento do capital, é
permeado de interesses econômicos que se intensificam ainda mais quando empresas
internacionais, como o Banco Mundial (BM) e o Fundo Monetário Internacional (FMI),
“passam a operar de forma articulada no contexto da globalização econômica internacional
para aprofundamento e a implementação de políticas que favoreçam a reprodução do capital”
(HADDAD, 2008, p. 7), intervindo junto aos governos por meio da imposição de temáticas
prioritárias e de uma abordagem economicista das políticas educacionais. Com olhares voltados para o cenário econômico brasileiro as organizações
internacionais adentram no campo educacional com o discurso de “investir para melhorar”,
porém utilizam-se da educação na lógica capitalista para acumular resultados positivos ao
mercado. A educação, nesse sentido, perde sua dimensão enquanto bem público e passa a
suprir as demandas das grandes empresas. Barroso (2005) salienta que: Na educação, se promovem, se discutem e se aplicam medidas políticas e
administrativas que vão, em geral, no sentido de alterar os modos de
regulação dos poderes públicos no sistema escolar (muitas vezes com
recurso a dispositivos de mercado), ou de substituir esses poderes públicos
por entidades privadas. (BARROSO, 2005, p. 726). As
orientações
das
políticas
educacionais
são
realizadas
sempre
em
complementaridade às orientações macroeconômicas estabelecidas, onde fica claro o interesse
em fazer reforma educacional a partir da lógica neoliberal e a favor da prevalência da
reprodução do grande capital. Dessa forma, pela ótica neoliberal a orientação é privatizar e
com isso retirar a liberdade de escolha e participação do Estado para dar conta das demandas
e necessidades do mercado. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. Relação estado-capital no delineamento das políticas educacionais Destarte a isso, as escolhas de políticas educacionais saem do poder do Estado e
passam a ser definidas pela “mão invisível” do mercado em função de objetivos definidos
pelo crescimento econômico. Para Barroso (2005) esse processo traduz-se, sobretudo, na
subordinação das políticas de educação a uma lógica estritamente econômica
(“globalização”); na importação de valores (competição, concorrência, excelência etc.) e
modelos de gestão empresarial, como referentes para a “modernização” do serviço público de
educação; na promoção de medidas tendentes à sua privatização. Nesta perspectiva, assiste-se à tentativa de “criar mercados (ou quase-mercados)
educativos transformando a ideia de ‘serviço público’ em ‘serviços para clientes’, onde o
‘bem comum educativo’ para todos é substituído por ‘bens’ diversos” (BARROSO, 2005, p. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. 382 Práxis
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Revista Revista e-ISSN 2178-2679 750). Logo, os produtos desse modelo geram a distribuição de serviços desiguais, uma
educação para o rico e outra para o pobre, não havendo problema de criar um organismo de
excluídos, pois seu objetivo central é adequar a educação a formar sujeitos para a demanda do
mercado, negligencia-se qualquer tipo de escolha que vise o bem comum. Em meio ao cenário da política neoliberal, a educação tornou-se um dos mais
importantes investimentos do cerne capitalista, isso implicou em mudanças, porém estas
negam os reais sentidos de uma sociedade igualitária e humanizadora, o que impera é a
hegemonia de um mercado que incentiva o lucro, a luta de classes, uma educação enquanto
mercadoria intensificando cada vez mais as premissas da teoria do capital humano
(FRIGOTTO, 2010). A educação passa a ser vista como um grande investimento e acaba sendo pretensão de
negociação entre os capitalistas, os quais passam a atribuir novas demandas educacionais
onde “os fatores sociais, políticos e econômicos que se estruturam a partir do processo de
mundialização do capital são determinantes para a configuração da área educacional que,
nesse contexto, tornou-se um grande mercado e o ensino uma mercadoria” (NASCIMENTO,
2011, p. 25). Relação estado-capital no delineamento das políticas educacionais Sendo assim, as políticas públicas educacionais constituídas a partir da manipulação
do capital acabam colocando o problema da educação nas “costas” dos sujeitos omitindo o
Estado dessa função, mas o certo é que muitas das medidas tomadas com relação às políticas
educativas favorecem a introdução de uma lógica de mercado na prestação do serviço
educativo, cujos efeitos contrariam claramente os princípios de uma formação de qualidade. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. 1 A ideia-chave é de que a um acréscimo marginal de instrução, treinamento e educação, corresponde um
acréscimo marginal de capacidade de produção. Ou seja, a ideia de capital humano é uma “quantidade” ou um
grau de educação e de qualificação, tomado como indicativo de um determinado volume de conhecimentos,
habilidades e atitudes adquiridas, que funcionam como potencializadoras da capacidade de trabalho e de
produção (FRIGOTTO, 2010, p. 44). A inserção da política brasileira de tecnologia no campo educacional Abordar a inserção das Tecnologias de Informação e Comunicação (TIC) na educação
pressupõe pensar, com base nos princípios da sociedade capitalista, qual seu papel no
processo de formação dos sujeitos, uma vez que, ao longo da história a educação é alçada
como instrumento de proliferação dos interesses de classe onde os processos educativos são
utilizados na produção e reprodução das relações sociais. Para isso, é necessário pensá-la a
partir do desenvolvimento histórico da sociedade capitalista reconhecendo as condições de
sua produção e compreendendo suas principais manifestações. A educação, ao longo do processo histórico da sociedade capitalista, é entendida
segundo Frigotto (2006) como algo não meramente instrumental, mas decisivo do ponto de Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. 383 Práxis
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Revista Revista e-ISSN 2178-2679 vista do desenvolvimento da economia. Nesse panorama, a teoria do capital humano1 se
configurou como um dos elementos constitutivos e reforçadores da tendência tecnicista de
educação no qual sua subordinação significa torná-la funcional ao sistema capitalista, isto é,
colocá-la a serviço dos interesses da classe dominante, uma vez que, qualificando a força de
trabalho, o processo educativo concorreria para o incremento da produção da mais-valia,
reforçando, em consequência, as relações de exploração. Deste modo, “a educação, quando apreendida no plano das determinações sociais e,
portanto, ela mesma constituída e constituinte destas relações, apresenta-se historicamente
como um campo de disputa hegemônica” (FRIGOTTO, 2010, p. 27) e passa a definir-se como
uma técnica de preparar recursos humanos para o processo de produção em prol dos interesses
de classes. Para Frigotto (2010) este cenário vislumbra duas vertentes antagônicas de formação
humana. Na perspectiva das classes dominantes a educação objetiva habilitá-los técnica,
social e ideologicamente para o trabalho a fim de responder as demandas do capital. Na
perspectiva dos grupos sociais, especialmente para a classe trabalhadora, a educação define-se
como o desenvolvimento de potencialidades que lhes permitam adquirir conhecimentos para
satisfazer seus interesses e necessidades. Ou seja, expõe a existência de uma escola dualista
em defesa de um ensino disciplinador e adestrador para os filhos dos trabalhadores e
formativo para os filhos das classes dirigentes. Isto evidencia que “a educação é o processo pelo qual a sociedade forma seus
membros à sua imagem e em função de seus interesses” (VIEIRA PINTO, 1993, p. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. A inserção da política brasileira de tecnologia no campo educacional 29),
assim, “a educação e a formação humana terão como sujeito definidor as necessidades, as
demandas do processo de acumulação de capital sob as diferentes formas históricas de
sociabilidade que assumir” (FRIGOTTO, 2010, p. 33). Portanto, a educação pode ser compreendida como uma arma de coerção usada pelo
capital, por via do Estado, para manter sua estrutura. É usada como meio de subordinação ao
mercado e a sua adaptabilidade e funcionalidade, quando na verdade deveria ser empregada
como elo mediador na formação de sujeitos mais críticos e reflexivos sobre suas reais
necessidades. A educação deveria promover a qualificação humana para o: 384 Práxis
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[...] desenvolvimento de condições físicas, mentais, afetivas, estéticas e
lúdicas do ser humano (condições omnilaterais) capazes de ampliar a
capacidade do trabalho na produção dos valores de uso em geral como
condição de satisfação das múltiplas necessidades do ser humano no seu
devenir histórico. (FRIGOTTO, 2010, p. 34). Revista e-ISSN 2178-2679 [...] desenvolvimento de condições físicas, mentais, afetivas, estéticas e
lúdicas do ser humano (condições omnilaterais) capazes de ampliar a
capacidade do trabalho na produção dos valores de uso em geral como
condição de satisfação das múltiplas necessidades do ser humano no seu
devenir histórico. (FRIGOTTO, 2010, p. 34). Porém, as práticas educativas ao serem subordinadas aos preceitos do capital acabam
adquirindo interesses diversos e antagônicos para as diferentes classes tendo como condição
indispensável a sua estruturação “a necessidade de que a reprodução da força de trabalho seja
moldada, forjada, fabricada para a disciplina e subordinação das novas relações de produção”
(FRIGOTTO, 2010, p. 36), assumindo-se como prática social de produção econômica, ou
seja, capital humano. A disseminação da teoria do capital humano é lançada como crença de mágica solução
das desigualdades sociais entre os países desenvolvidos e os subdesenvolvidos representando
dominantemente a visão e os interesses do capitalismo integrado ao grande capital. Isso
perdura ao longo da histórica relação entre trabalho-educação e se intensifica a partir da
década de 90 quando é incorporada ao debate da tecnologia. O surgimento das novas tecnologias permite identificar uma problemática que se
expõe como desafio aos processos educativos, pois estas se inserem ocasionando mudanças
sobre o trabalho e a qualificação humana e refletem, na maioria das vezes, o ideário da
sociedade capitalista, o qual acaba por assumir novas formas e demandas oriundas da lógica
do mercado. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. A inserção da política brasileira de tecnologia no campo educacional Este desafio se apresenta no plano político-ideológico denominado de Sociedade
da Informação ou do Conhecimento (dentre outros inúmeros termos usados) ambas
embasadas no acesso “global” a informação, conhecimento e desenvolvimento. A expressão “Sociedade da Informação” surge no contexto da pós-modernidade, mais
precisamente na década de 1970, a partir das discussões sobre o que seria a “sociedade pós-
industrial” e juntamente com o termo “globalização” apresentando novas condições para o
processamento de informação. Essa reformulação social não passa de uma reorganização do
capital em meio as suas crises, por vezes cíclicas, na busca de instabilidade financeira e tem
como atributos chaves a informação e o conhecimento enquanto mecanismos de promoção do
desenvolvimento técnico-científico. Para Pimenta (2014) as origens e causas desta sociedade se alicerçam em dois tipos de
desenvolvimento interdependentes: o desenvolvimento econômico a longo prazo e a mudança
tecnológica. Ambos estão estritamente ligados à reestruturação do capital para o qual as novas
tecnologias de informação e comunicação surgem como instrumentos possibilitadores do
processo de globalização e ampliação do sistema. 385 Práxis
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negócios e, por consequência, da economia, da sociedade e da política, de acordo com
Pimenta (2014). Dentro dessa ordem econômico-social, sob a égide da Sociedade da
Informação, defende-se a busca por uma nova qualidade da educação e a formação com
princípios alçados aos requisitos e demandas do mercado para o qual é fundamental uma
eficiente qualificação para o trabalho que vislumbre “uma formação geral e abstrata, que
prepara sujeitos polivalentes, flexíveis e participativos” (FRIGOTTO 2010, p. 59). A exigência dessa sociedade é que: A informação se traduza em ativo estratégico, diferencial competitivo,
recurso gerencial e vetor de desenvolvimento. E esta exigência se sustenta
no uso intensivo das tecnologias de informação e comunicação, que
prometem eliminar os marcadores de fronteiras entre os conectados e os
desconectados, com a mesma ênfase que se propõem a eliminar as barreiras
entre a ignorância e o saber. (PIMENTA, 2014, p. 38). Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. democratização do acesso à informação e a produção de conhecimentos reduzidos à questão
econômica e guiados pela ênfase nas habilidades e competências para o mercado de trabalho. democratização do acesso à informação e a produção de conhecimentos reduzidos à questão
econômica e guiados pela ênfase nas habilidades e competências para o mercado de trabalho. Considerando a relação orgânica entre Estado e capital, no conjunto das
políticas neoliberais, o primeiro usa o aparato tecnológico como um
mecanismo ideológico, para implementar o ajuste estrutural impostos pelos
organismos internacionais e o segundo o faz com a finalidade de recompor
suas taxas de acumulação, garantir o controle do processo produtivo e
explorar novos nichos de mercado (NASCIMENTO, 2011, p. 30-31). Neste cenário o “Estado viu-se obrigado a se adequar ao novo paradigma técnico e
econômico, à liberalização dos mercados nacionais e à “globalização” da economia, passando,
então, a assumir um papel redimensionado nos planos produtivo e administrativo”
(PIMENTA, 2014, p. 40). Assim, o Brasil, fortemente envolvido pela bolha da
“globalização”, no início dos anos 1990, passou a adotar políticas neoliberais, com base na
flexibilização do mercado. O Brasil para se inserir nesse novo panorama socioeconômico da Sociedade da
Informação apresenta o Programa Sociedade da Informação - SOCINFO, publicado no Livro
Verde, no ano de 2000 apoiando-se no novo paradigma de informação como bem econômico
e de informação estratégica para o desenvolvimento (PIMENTA, 2014). O SOCINFO é na
verdade a primeira iniciativa do governo brasileiro em definir um projeto estratégico que
pudesse integrar e coordenar o desenvolvimento e a utilização de serviços avançados de
computação, comunicação e informação. O Brasil para se inserir nesse novo panorama socioeconômico da Sociedade da
Informação apresenta o Programa Sociedade da Informação - SOCINFO, publicado no Livro
Verde, no ano de 2000 apoiando-se no novo paradigma de informação como bem econômico
e de informação estratégica para o desenvolvimento (PIMENTA, 2014). O SOCINFO é na
verdade a primeira iniciativa do governo brasileiro em definir um projeto estratégico que
pudesse integrar e coordenar o desenvolvimento e a utilização de serviços avançados de
computação, comunicação e informação. Os Livros Verdes são, pois, o resultado da discussão de iniciativas mundiais voltadas
para a construção de uma sociedade da informação para qual o advento é o fundamento de
novas formas de organização e de produção em escala mundial, redefinindo a inserção dos
países na sociedade internacional e no sistema econômico mundial. Segundo este documento
caberia ao sistema político promover políticas de inclusão social, para que o salto tecnológico
tenha paralelo quantitativo e qualitativo nas dimensões humana, ética e econômica
(TAKAHASHI, 2000). Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. A inserção da política brasileira de tecnologia no campo educacional No bojo da reestruturação produtiva do capital, as novas tecnologias de informação e
comunicação surgem produzindo e acelerando os processos comunicativos e com a
perspectiva de gerar ou transformar informação em conhecimento, em prol do aumento da
produtividade do sistema econômico, pois este a priori é compreendido como o principal fator
de produção técnico-científica e produtor de desenvolvimento. Dessa forma, este paradigma
social deixa claro que se trata “simplesmente de uma nova fase do capitalismo, em que as
tecnologias de informação adquirem uma relevância fundamental” (BOLAÑO, 2000, p. 127),
atribuindo caracteres utilitaristas a informação e ao conhecimento sem perder sua coerência
interna de busca permanente por expansão, acumulação e concentração. Para Mészáros (2011), o sistema capitalista é a mais poderosa estrutura “totalizadora”
de controle onde tudo deve se ajustar a ela, inclusive os seres humanos, e provar sua
viabilidade produtiva. O autor destaca este sistema como algo globalmente dominante que se
sobrepõe a tudo e a seus próprios critérios de viabilidade, sempre a favor do grande capital. É
com base nesse argumento que este caracteriza o sistema como um movimento incontrolável
“totalizador irrecusável e irresistível, não importa quão repressiva tenha de ser a imposição de
sua função totalizadora” (MÉSZÁROS, 2011, p. 96). O Brasil, por não está fora do contexto massacrante do capitalismo e da
“globalização”, com sua economia presa a um círculo vicioso, vive administrando sempre
tensas e paradoxais relações entre a mão pesada do Estado e a “mão invisível do mercado”
(PIMENTA, 2014, p. 47), que a partir da perspectiva neoliberal acaba por disseminar a 386 Práxis
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democratização do acesso à informação e a produção de conhecimentos reduzidos à questão
econômica e guiados pela ênfase nas habilidades e competências para o mercado de trabalho. Revista e-ISSN 2178-2679 democratização do acesso à informação e a produção de conhecimentos reduzidos à questão
econômica e guiados pela ênfase nas habilidades e competências para o mercado de trabalho. democratização do acesso à informação e a produção de conhecimentos reduzidos à questão
econômica e guiados pela ênfase nas habilidades e competências para o mercado de trabalho. A chamada “alfabetização digital” é elemento-chave nesse quadro. Porém, isso ocorre
somente no papel, pois o que se vê, principalmente nos países periféricos, é o aumento intenso
da exclusão digital ocasionada pela tão propagada “globalização”, uma vez que, apesar de se
apresentar como um acontecimento muito benéfico ao desenvolvimento social dos países,
esse fenômeno surge sobre o domínio do capital provocando mudanças em prol dos interesses
dos países desenvolvidos deixando os demais à mercê das desigualdades e da exploração. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. 387 Práxis
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divulgada sociedade, por ser esta uma ideologia produzida pelo capitalismo e por preservar o
modelo de exclusão, que caminha no sentido oposto ao da proposta de incluir a todos. Assim,
para esta autora a inclusão digital se apresenta como uma “ilusão”, pois é estruturada com o
propósito de atender às demandas do mercado capitalista na qual as políticas adotadas nessa
direção apenas acrescentam pequenas mudanças, não intervindo na estrutura mais ampla, e
contribuindo com a reprodução da estrutura de dominação e legitimação de valores
dominantes. Dessa forma, as políticas de inserção das tecnologias na educação seguem da trama
dos preceitos neoliberais do Estado capitalista na qual a “dinâmica estrutural da sociedade
capitalista é determinante na formulação de concepções e orientações para o ensino”
(NASCIMENTO, 2011, p. 25). Para este autor, o uso das tecnologias de informação e
comunicação, em um cenário de novas demandas educacionais, determinadas pela
mundialização do capital, está acompanhado da própria noção do que se deseja educar, de
qual é a função das instituições de ensino e de que sujeito se pretende formar. Nesta instância “as políticas públicas sociais caminham no sentido de manutenção da
ordem estabelecida, uma vez que não rompem com as estruturas de reprodução das
desigualdades” (PRAZERES, 2016, p. 62), na qual o Programa Brasileiro da Sociedade da
Informação se “apresentou como mais um projeto capitalista de ajuste do Estado brasileiro às
pressões dos países hegemônicos, cujo principal objetivo era inserir o país na economia
mundial globalizada” (PIMENTA, 2014, p. 9). Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019 democratização do acesso à informação e a produção de conhecimentos reduzidos à questão
econômica e guiados pela ênfase nas habilidades e competências para o mercado de trabalho. Deste modo, Nascimento (2011) afirma que: [...] a sociedade do conhecimento e sua sociabilidade pautada nas novas
tecnologias de informação e comunicação, não representam uma ruptura da
configuração capitalista, não delineia novas relações classistas, mas tem
como objetivo principal operar ideologicamente na construção do fetichismo
tecnológico que reconfigura o modus operandi da produção capitalista
industrial e estabelece uma nova era social e educacional centrada nas
possibilidades de construção de uma cidadania ‘global’, ‘em rede’,
viabilizada pelo poder do acesso à informação e pela possibilidade
imensurável de comunicação. (NASCIMENTO, 2011, p. 69). Nesta perspectiva, para este autor, há a inserção de uma noção fetichista de tecnologia
engendrada pelo Estado e determinada pelo capital que articula o uso da inovação tecnológica
às suas finalidades utilizando-a como preceito de formação para necessária inserção no Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. 388 Práxis
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mundo do trabalho. Ou seja, tomam a tecnologia como uma variável, um fator independente e
autônomo aos interesses de classe e às relações de poder. Práxis
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Revista Revista e-ISSN 2178-2679 mundo do trabalho. Ou seja, tomam a tecnologia como uma variável, um fator independente e
autônomo aos interesses de classe e às relações de poder. mundo do trabalho. Ou seja, tomam a tecnologia como uma variável, um fator independente e
autônomo aos interesses de classe e às relações de poder. O autor acentua ainda que a sociabilidade capitalista ancorada nas novas tecnologias e
no fetichismo tecnológico determinados pelo capital em um contexto geral, no nível das
relações sociais e de produção, se materializa em contextos particulares, como no caso das
políticas públicas educacionais, e é determinada por um fenômeno muito mais complexo, o da
reestruturação do processo de acumulação capitalista para fins de reprodução do capital. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. Considerações finais A relação entre a sociedade e a tecnologia não se apresenta de forma tranquila, isso
por que, apesar dos avanços obtidos pelas sucessivas revoluções tecnológicas, não foi possível
garantir melhoria de qualidade de vida a amplos segmentos sociais que ainda se encontram
excluídos, vivendo às margens dos bens sociais e tecnológicos. Assim, é preciso estudar esse
impacto sem perder de vista todos os efeitos causados por essa interatividade. De tal modo, infelizmente o acesso aos bens tecnológicos, ainda, é restrito e não
garante a inclusão de todos, pois, a possibilidade de acesso tecnológico trava no embate
riqueza-pobreza. O desafio do desenvolvimento tecnológico depende do potencial de
desenvolvimento socioeconômico, das lutas individuais e coletivas pelo acesso igualitário aos
bens sociais e tecnológicos. Em nosso país, lamentavelmente, o que se observa é uma injusta
distribuição de renda que influência diretamente no acesso aos recursos tecnológicos, apesar
da tão propagada defesa da introdução das tecnologias em todos os setores públicos. Sob essa perspectiva, as políticas de educação, pensadas para incluir digitalmente
segmentos marginalizados da sociedade, são insuficientes, pois inserem insumos tecnológicos
na escola priorizando aspectos meramente técnicos ou tecnológicos. Assim, argumenta-se a
necessidade dessas políticas serem repensadas para proporcionar uma formação que não
preconize somente demandas do mercado, mas ações que desenvolvam a formação integral do
indivíduo e ao mesmo tempo estejam acima das lutas de classes e dos interesses do mercado
para que sejam destinadas aos interesses do indivíduo e da sociedade. Alertamos, a partir de Vieira Pinto (2005), que a tecnologia não deve ser
compreendida como causa, mas mediação, onde as forças de ascensão nos países pobres
devem tomar consciência e de que precisam lançar mão para lutar contra velhas estruturas de
relações sociais. Paradoxalmente, exaltar a tecnologia significaria retardar seu efeito
libertador a ponto de fazer dela uma nova mitologia, com sua numerosa procissão de Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. 389 Práxis
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Revista Revista e-ISSN 2178-2679 idólatras. A tecnologia, desse modo, tem que ser um meio para nossas relações dentro do
modelo capitalista, para transformá-lo, e não para mantê-lo. Considerações finais A análise do autor sobre as relações entre interesses econômicos e determinado padrão
tecnológico aponta para a necessidade de se estar atento, no campo pedagógico, ao fato de
que, ao se copiar acriticamente os projetos e modelos adotados no campo industrial, interesses
e lógicas próprias estão sendo incorporados. Nem sempre a lógica da indústria é a mesma da
educação, os interesses podem ser diferentes e, fundamentalmente, a natureza dos resultados e
o tempo em que eles ocorrem podem ser diferenciados. Nessa leitura, toda tecnologia consiste
em uma determinada concepção do significado e do valor das ações humanas. Logo, a educação que interessa à classe trabalhadora não é a que inseri insumos
tecnológicos para o uso técnico da tecnologia, mas aquela que possibilita a reflexão, que
permite o sujeito se enxergar enquanto produtor de valor, que politiza, que desaliena, que
forma para a vida em qualquer sociedade. Essa proposta ainda não se encontra em nenhuma
estrutura de ensino pensada pelo Estado, considerando seu caráter neoliberal, encontra-se nos
movimentos sociais, nas comunidades de base, nos partidos políticos e nas práticas
pedagógicas de professores comprometidos com a transformação social. REFERÊNCIAS BALL, S. J. Performatividade, privatização e o estado do bem estar. Educação &
Sociedade, v. 25, no. 89, PP. 1105-1126, dez. 2004. BARROSO, João. O Estado, a educação e a regulação das políticas públicas. IN:
Educação e Sociedade, Campinas, vol. 26, n. 92, p. 725-751, Especial, Out, 2005, p. 725- BOLAÑO, César Ricardo Siqueira. Sociedade da informação: reestruturação capitalista e
esfera pública global. Araraquera/SP, v. 8, n. 00, p. 93-128, 2000. Disponível em
<https://seer.fclar.unesp.br/estudos/article/viewFile/834/695> Acesso 19 jul. 2017 às
17h45min. CASTELLS, Manuel. A sociedade em rede. Vol. 1. São Paulo: Paz e Terra, 1999. SOBRE AS AUTORAS Maria Sueli Corrêa dos Prazeres Doutora em Educação pela Universidade Estadual de Ponta Grossa (UEPG). Docente do
Programa de Pós-graduação em Educação e Cultura, da Universidade Federal do Pará
(UFPA). Membro associado da Red Latinoamericana de Estudios Epistemológicos en Política
Educativa (ReLePe). E-mail: suelicorrea@ufpa.br CASTELLS, Manuel. A sociedade em rede. Vol. 1. São Paulo: Paz e Terra, 1999. FRIGOTTO, Gaudêncio. A produtividade da escola improdutiva: um (re) exame das
relações entre educação e estrutura econômico-social capitalista. 8ª edição. São Paulo: Cortez,
2006. FRIGOTTO, Gaudêncio. Educação e a crise do capitalismo real. 6ª edição. São Paulo:
Cortez, 2010. Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. 390 Práxis
Educacional
Revista Revista e-ISSN 2178-2679 HADDAD, Sérgio (Org.) Banco Mundial, OMC e FMI. O Impacto nas Políticas
Educacionais. São Paulo: Cortez Editora, 2008. MÉSZÁROS, István. A educação para além do capital. 2ª Edição. Tradução de Isa Tavares. São Paulo: Boitempo, 2008. MÉSZÁROS, István. Para além do capital: Rumo a uma teoria da transição. Tradução de
Paulo Cezar Castanheira e Sérgio Lessa. São Paulo: Boitempo, 2011. NASCIMENTO, Alberico Francisco. A Educação a Distância e fetichismo tecnológico:
Estado e Capital no projeto de Ensino Superior no Brasil. 2011. 233 f. Tese (Doutorado em
Políticas Públicas) – Universidade Federal do Maranhão, São Luís, 2011. PIMENTA, Márcia Teresa da Rocha. A política de inserção do Brasil na “Sociedade da
informação”: uma avaliação política do Programa Sociedade da Informação – SOCINFO. 2014. 222 f. Tese (Doutorado em Políticas Públicas) – Universidade Federal do Maranhão,
São Luís, 2014. PRAZERES, Maria Sueli Corrêa dos. O Programa Navega Pará como política pública de
inclusão digital: implicações nas escolas públicas do Estado Pará, 2016. 274 fls. Tese
(Doutorado em Educação) – Universidade Estadual de Ponta Grossa, Ponta Grossa, 2016. TAKAHASHI, Tadeu (Org.). Sociedade da Informação no Brasil: Livro Verde. Brasília:
Ministério da Ciência e da Tecnologia, 2000. VIEIRA PINTO, Álvaro. Sete lições sobre educação de adultos. 8ª Edição. São Paulo:
Cortez, 1993. VIEIRA PINTO, Álvaro. Sete lições sobre educação de adultos. 8ª Edição. São Paulo:
Cortez, 1993. VIEIRA PINTO, Álvaro. O conceito de tecnologia. Vol. 1. São Paulo: Contraponto, 2005. VIEIRA PINTO, Álvaro. O conceito de tecnologia. Vol. 1. São Paulo: Contraponto, 2005. Ilda Gonçalves Batista Mestre em Educação e Cultura pela Universidade Federal do Pará (UFPA). Professora da
Rede Municipal de Parauapebas. Membro associado no Grupo de Pesquisa Trabalho,
Tecnologia e Educação do Campo. E-mail: ildagoncalves92@yahoo.com.br Recebido em: 05 de abril de 2018
Aprovado em: 16 de agosto de 2018
Publicado em: 10 de maio de 2019 Revista Práxis Educacional, Vitória da Conquista – Bahia – Brasil, v. 15, n. 32, p. 378-391, abr./jun. 2019. 391
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On Komatsu’s Strategy of Distribution Channels in China ——Take Komatsu Excavators as an Example
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Sufang Zhang
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International Journal of Marketing Studies International Journal of Marketing Studies www.ccsenet.org/ijms 1. Introduction Nowadays, the competition of MNCs in the global market focuses on two areas: one is the field of product
research and development; the other is the field of product distribution. The former is to grab dominated
technology position, while the latter is to grab dominated market position. Technologically superior products
may not necessarily have a superiority in the market. How to transform the superority in the field of technology
to the superiority in the market is a great concern of MNCs. Distribution channel, also known as marketing channel, is the flow channel of products from the producer to the
consumer. It consists of the producer, the consumer, and various intermediaries between them. The function of
distribution channel is to eliminate asymmetry of information between the producers and consumers so that
transactions can be realized. The study of distribution channel concentrates on selection and management of
distribution channels, which is the basic research area of marketing theory. In recent years, studies have shown
that selection and management of distribution channels have been not only the management functions and daily
operations of MNCs, but also an important part in acquiring core competencies and competitiveness advantage
for MNCs. Since the reform and opening up, more and more MNCs have entered the Chinese market through a variety of
forms. By virtue of their sophisticated marketing experience and creative marketing idea, they have created
miracles in many markets. Japan Komatsu Co., Ltd. (hereinafter called Komatsu) is one of them. Komatsu
started its production of excavators in China in 1995 in the form of joint ventures in China. The sales of its
excavators in China in 2006 reached more than 6,000 units, taking a market share of 16.7%. There are many
reasons for Komatsu’s success in the Chinese market, but a successful distribution channel strategy, is
undoubtedly one of the key factors that help to expand their scale of production. By analyzing the distribution
channel of Komatsu’s excavators in China, this paper explores the strategy of MNCs in obtaining competitive
advantages and provides a reference for cross-border operations for Chinese enterprises. Abstract Selection and management of distribution channels are not only part of management functions and daily
operation of multinational corporations (MNCs), but also important compositions of core capability and
competitive advantages. This paper first analyzes Komatsu, a well-known Japanese company’s strategy of
distribution channel of excavators in China from the perspective of distribution channel intensity, then it
discusses market function positioning of Komatsu’s distributors and Komatsu’s control of its distribution
channels. Thirdly the paper summarizes characteristics of Komatsu’s distribution channels and conduct
theoretical thinking on the strategy of distribution channels of MNCs. Finally it suggests that Chinese enterprises
learn from the successful experience of Komatsu. Keywords: MNCs, Komatsu, Excavators, Distribution Channel On Komatsu’s Strategy of Distribution Channels in China
——Take Komatsu Excavators as an Example Sufang Zhang
School of Economics and Management, Northern China Electric Power University
#2 Beinong Road Deshengmenwai, Beijing 100022, China
E-mail: zsf69826313@sina.com.cn Chenwei Fu
School of Economics and Management, Northern China Electric Power University
#2 Beinong Road Deshengmenwai, Beijing 100022, China
E-mail: chin_hans@yahoo.com.cn nurtured and reconstructed China's domestic channel system.
2.1 Intensity of distribution channel nurtured and reconstructed China's domestic channel system. 2.1 Intensity of distribution channel At present, the Komatsu excavator network of distribution channels in China covers the country's 31 provinces,
municipalities and autonomous regions. There are 34 distributors and 238 affiliated subsidiaries, branches and
offices. The channel division strictly follows the basic principle of regional governance, with an average of 0.94
distributors for each province or municipality and an average of 7.21 branches for each distributor. There may be
two distributors being designated in the same province such as Shangdong, Hunan and Sichuan due to intensive
users or scarce distribution outlets in these places, while two provinces or autonomous regions such as Gansu,
Ningxia may share one distributor because of their user scarceness. Such a distribution density can both ensure
that the users of evacuators in each province obtain products and services within a relatively short period and
that each distributor has sufficient source of income to maintain market operation and promotional costs, and
obtain excess profits and excess rewards given by Komatsu for exceeding annual sales target. In addition, it
avoids vicious price competition or market disorder. 2.2 Positioning of distributors with respect to market functions (9) Maintenain the power of implementation of sales process, operating capacity of salesman and service
personnel's technical grade, accept regular training and evaluation required by Komatsu. (10) Conduct transactions directly with end consumers in accordance with the market price system and the
discount rate, provide products and services according to Komatsu standards and do not cut prices in excess of
authority. (11) Make great efforts to recover the purchase price. High-risk tranactions such as payment by installment are
not allowed without Komatsu permission. The channel system does not include dealers. Products are to be sold to the dealers at the average prices to the
customers, or distributors are not allowed to offer discounts which are above the standard prices. IIn a rapid
growing market like China, it is a wise approach to compress the channel length to the minimum to ensure
maximum profits. In addition, no direct sales system is established under this channel system, reflecting
Komatsu’s sufficient trust in its distributors. 2. Strategy of distribution channel of Komatsu excavator products in China Komatsu's success in the Chinese market is inseparable from the success of establishing its distribution system in
China. Since 2001, Komatsu, with its rich experience of international marketing channel construction, have 254 International Journal of Marketing Studies Vol. 2, No. 1; May 2010 2.2 Positioning of distributors with respect to market functions (1) Distributor responsibility system is based on division of market region. Selling and providing services
beyond agreed areas are absolutely prohibited. 1) Distributor responsibility system is based on division of market region. Selling and providin
beyond agreed areas are absolutely prohibited. butors in different regions are legal units independent from each other. They are autonomous
ing and taking the financial risk for failing to operate according to Komatsu operational procedures. 2) Distributors in different regions are legal units independent from each other. They are au
elf-financing and taking the financial risk for failing to operate according to Komatsu operational pro (3) Distributors should maintain inventory quantity, variety and quality of evacuators and spare parts authorized
by Komatsu. (4) Distributors must accept Komatsu's sales and technical training; establish qualified management standards
under the supervision of market representatives and service representatives sent by Komatsu, apply for support
from Komatsu for visits to key customer and accept suggestions with regard to inventory vehicles and parts, deal
with process-type supervision and management such as report regularly about sales information and financial
reconciliation. (5) Qualified distributor should strictly perform their market functions authorized by Komatsu such as promotion,
sales, service and support. (5) Qualified distributor should strictly perform their market functions authorized by Komatsu such as promotion,
sales, service and support. (6) Distributors must promise not to operate products of Komatsu’s rivals, with the exception of non-competitive
products. (6) Distributors must promise not to operate products of Komatsu’s rivals, with the exception of non-competitive
products. (7) Regional or national distributors accept the arbitration of Komatsu's Market Supervision Corporation in the
event of internal conflict between them. (7) Regional or national distributors accept the arbitration of Komatsu's Market Supervision Corporation in the
event of internal conflict between them. (8) Distributors who succeed in performing their market functions shall be provided additional bonuses and
promotion by Komatsu; unqualified distributors shall be punished by disgrading credit, deducting bonuses,even
being deprived of qualification. (8) Distributors who succeed in performing their market functions shall be provided additional bonuses and
promotion by Komatsu; unqualified distributors shall be punished by disgrading credit, deducting bonuses,even
being deprived of qualification. (9) Maintenain the power of implementation of sales process, operating capacity of salesman and service
personnel's technical grade, accept regular training and evaluation required by Komatsu. 3.1 Reward For the purpose of raising the market share, Komatsu sets the target of sales units for each distributor and give
rewards to the distributors from the collected payment of goods. 1) As noted above, rewards to those distributors who fail to fulfil their sales target and defer payment to Komatsu without any excuses will be deducted at a certain rate. If distributors do not realize the sales and
market share targets for a relatively long period, their selling rights will be cancelled. o Komatsu without any excuses will be deducted at a certain rate. If distributors do not realize the
market share targets for a relatively long period, their selling rights will be cancelled. (2) The implementation of distributors star rating system. Under this system, if the distributors meet the specific
reqruiements of stnadard in terms of warehouse building, special tools and vehicles in place, staff enrichment,
training in place, the number and types of spare parts etc., the distributors will be promoted in terms of reward
quotas and training levels. (2) The implementation of distributors star rating system. Under this system, if the distributors meet the specific
reqruiements of stnadard in terms of warehouse building, special tools and vehicles in place, staff enrichment,
training in place, the number and types of spare parts etc., the distributors will be promoted in terms of reward
quotas and training levels. (3) Adopt a common approach in developed countries, require all distributors use an Internet-based distributor
database management system and IT technology to enable the work of distributors networking, time-oriented,
process-oriented with respect to purchase, inventory, reporting, reimbursement, sales, bad debts, reward, spare
parts, service, claims, etc. the purpose of which is to strengthen the management of distribution channels and
improve efficiency. 4.4 The approach of fostering and nurturing distributors Distributors are required to implement the strict management measures and Komatsu often sends staff to carry
out real-time monitoring. The marketing capability and management level of the distributors are thus promoted,
which is conducive to sustained and long-term distribution operation and to the maintainance of stable
distribution channel. 3.3 Price Komatsu sets a unified market retail price for distributors and strict rules of discounts enjoyed by distributors at
different levels. It not only determines the price and discount level, but also make stringent regulations on
standards for fee collection with regard to freight, renovation services, supply levels of spare parts, repairs and
minor repairs and claim standards, etc. International Journal of Marketing Studies www.ccsenet.org/ijms 3.1 Reward
For the purpose of raising the market share, Komatsu sets the target of sales units for each distributor and give
rewards to the distributors from the collected payment of goods. 3.2 Constraint
(1) As noted above, rewards to those distributors who fail to fulfil their sales target and defer payment
to Komatsu without any excuses will be deducted at a certain rate. If distributors do not realize the sales and
market share targets for a relatively long period, their selling rights will be cancelled. (2) The implementation of distributors star rating system. Under this system, if the distributors meet the specific
reqruiements of stnadard in terms of warehouse building, special tools and vehicles in place, staff enrichment,
training in place, the number and types of spare parts etc., the distributors will be promoted in terms of reward
quotas and training levels. (3) Adopt a common approach in developed countries, require all distributors use an Internet-based distributor
database management system and IT technology to enable the work of distributors networking, time-oriented,
process-oriented with respect to purchase, inventory, reporting, reimbursement, sales, bad debts, reward, spare
parts, service, claims, etc. the purpose of which is to strengthen the management of distribution channels and
improve efficiency. 3.3 Price
Komatsu sets a unified market retail price for distributors and strict rules of discounts enjoyed by distributors at
different levels. It not only determines the price and discount level, but also make stringent regulations on
standards for fee collection with regard to freight, renovation services, supply levels of spare parts, repairs and
minor repairs and claim standards, etc. 3.4 Leagal rights
Komatsu has review and approval rights with regard to the use of core technology and intellectual property right 3. Control of distribution channels In the more and more competitve Chinese market, differences of products and sevices between competitors has
been increasingly small and marketing tends to be homogenous. Therefore, being in firm control of distribution
channels is the sole pathway to obtain adequate profits. Komatsu exercises its control of distribution channels,
mainly from the aspects of reward, constraint, price and legal rights. 255 International Journal of Marketing Studies 4.1 Wide distribution channel Komatsu has a number of intensive distributors for selling excavators in China which in turn provides wide
channel coverage and makes its goal of penetration into a broader regional segment markets and acquiring a
higher market share possible. Of course, such a wide-channel also means increased workload for Komatsu in
coordinating channel operation and resolving channel conflict issues. 4.2 Reasonable geographical division of the channel Reasonable geographical division of the channel avoids unnecessary conflicts between channel members. Sufficient market area provides distributors with sufficient client resources, the principle of " more pay for more
work" encourages distributors in opening up markets as well as efforts in increasing market share. 4.3 Localization advantage of the selected distributors Local distributors are familiar with the local market conditions, cultural customs and consumer psychology. Thus,
they have unique advantages with respect to project operation and customer’s relationship. Meanwhile,
specializing in Komatsu excavators increases distributors concentration and is conducive to promote their
professional quality. 4.4 The approach of fostering and nurturing distributors 3.4 Leagal rights Komatsu has review and approval rights with regard to the use of core technology and intellectual property right,
information reported, summary, disclosure, communication, etc. In additon, it established stringent requirements
for the use of intangible assets. Komatsu expressly authorizes distributors at all levels. Act of the distributors
beyond their authority is strictly prohibited. 6. Concluding remarks Along with the thorough promotion of China's "marching out" strategy in the recent decade, more and
more Chinese enterprises have been entering international market, but compared with the MNCs of
developed countries, Chinese enterprises lack experience in cross-border investment. It is therefore
significant for them to learn from the successful experience of MNCs of developed countries. It is hoped
that the strategy of Komatsu excavator distribution channels in China can provide some useful lessons for
Chinese enterprises. 5.1 The design concept of the value chain of Komatsu distribution channels According to Michael Porter's elaboration of the synergistic effect of value chain, activities within the enterprise
can be decomposed of a number of components from the perspective of the strategic importance and their
combination will create value. Extended studies of the theory have shown that the value chain of a single
enterprise can be decomposed and integrated to form an external collaboration system consisting of a number of
independent members with different functions. From the analysis of the whole value chain, large-sized
manufacturing enterprises may give up some value added segments and design their own value chain system in
order to seek partners who will be cultivated to increase their core competitiveness and jointly complete the
whole process of value chain. The value chain of distribution channel of construction machinery is composed of manufacturers, distributors or
dealers/ retailers and end consumers. This paper argues that, manufacturers and end consumers who are in the
two ends of the value chain both have the initiative for interests. Through the market segmentation and target
market positioning, manufacturers may determine the end-user groups, end-user groups can also choose different
suppliers according to their own interests at different time. In promoting co-operation between the two, and
formally starting realization of transformation of value with each other, distribution channels between them play
a vital role. 5.2 The synergy effect of distribution channel members and enhancement of performance n implementing distribution channel strategy, it is a critical path to study channel relationships and e
o build and deal with channel relationships. When Komatsu first entered Chinese market, Komatsu faced the problem of the lack of distribution channels in
China's domestic market. By the virtue of its global experience in distribution channel construction, Komatsu
adopted the strategy of strict management together with careful nurture of distribution channels, creating
distribution channels of its own characteristics suitable for excavators in China. Strict management enables
domestic distributors make substantial progress in professional quality. Careful nurture helps Komatsu build
appropriate channel system aimed at acquiring market-share. The domestic channels, after experiencing a severe
test, also acquire mature marketing management experience and operational skills. Such inter-organizational collaboration and cooperation undoubtedly have raised the stability of the system and
increased mutual trust. The construction in IT, channel performance appraisal and hardware construction has also
benefited distributors. The overall efficiency of the channels has been improved substantially. 4.5 Attach importance to the communication with distributors Komatsu has a high degree of management localization in China. Many senior managers are mainland-born
Chinese who are concerned with the domestic market. They give enthusiastic support to the development of 256 International Journal of Marketing Studies Vol. 2, No. 1; May 2010 distributors and pay regular visits to distributors. They adopt the method of repeated consultations when
encountering serious problems rather than taking a simple confrontation approach in dealing with confrontation. distributors and pay regular visits to distributors. They adopt the method of repeated consultations when
encountering serious problems rather than taking a simple confrontation approach in dealing with confrontation. Xue Qiuzhi & Xia Kejia. (1999). New Concept of Marketing Distribution Channel of MNCs in China. Jounral
of International Trade Issue, (10). Fu Shali. (2008). Annalysis on Innovation and incentives of MNCs’Marketing Distribution Channel in China.
Journal of Commerce Study, (2): 209-211.
Niu Baoquan. (2008). Distribtuion Coopeation Theory and Application. Journal of Commerce Study, (1):
115-118.
Rosenbloom, B. (1999). Marketing Channels:A Management View (6th ed.). T.X: Dryden Press: 219.
Stern, L.w., and El-Ansart, A.T. (1996). Marketing Channels, Prentice Hall, Inc.
Xue Qiuzhi & Xia Kejia. (1999). New Concept of Marketing Distribution Channel of MNCs in China. Jounral
of International Trade Issue, (10). Journal of Commerce Study, (2): 209-211.
Niu Baoquan. (2008). Distribtuion Coopeation Theory and Application. Journal of Commerce Study, (1):
115-118. Niu Baoquan. (2008). Distribtuion Coopeation Theory and Application. Journal of Commerce Study, (1):
115-118. Fu Shali. (2008). Annalysis on Innovation and incentives of MNCs’Marketing Distribution Channel in China.
Journal of Commerce Study, (2): 209-211. Rosenbloom, B. (1999). Marketing Channels:A Management View (6th ed.). T.X: Dryden Press: 219. References Fu Shali. (2008). Annalysis on Innovation and incentives of MNCs’Marketing Distribution Channel in China. Journal of Commerce Study, (2): 209-211. Niu Baoquan. (2008). Distribtuion Coopeation Theory and Application. Journal of Commerce Study, (1):
115-118. Rosenbloom, B. (1999). Marketing Channels:A Management View (6th ed.). T.X: Dryden Press: 219 Rosenbloom, B. (1999). Marketing Channels:A Management View (6th ed.). T.X: Dryden Press: 219. Stern, L.w., and El-Ansart, A.T. (1996). Marketing Channels, Prentice Hall, Inc. 257
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Redetermination of<i>trans</i>-cyclohexane-1,4-diammonium dichloride
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Guido J. Reiß
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English
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organic compounds Experimental
Crystal data
C6H16N2
2+2Cl
Mr = 187.11
Monoclinic, P21=n
a = 5.2550 (11) A˚
b = 14.890 (3) A˚
c = 6.3604 (12) A˚
= 99.824 (18)
V = 490.39 (16) A˚ 3
Z = 2
Mo K radiation
= 0.60 mm1
T = 293 (2) K
0.30 0.24 0.20 mm
Data collection
Stoe STADI CCD diffractometer
Absorption correction: none
13502 measured reflections
1766 independent reflections
1562 reflections with I > 2(I)
Rint = 0.043
Refinement
R[F 2 > 2(F 2)] = 0.038
wR(F 2) = 0.072
S = 1.03
1766 reflections
79 parameters
All H-atom parameters refined
max = 0.41 e A˚ 3
min = 0.28 e A˚ 3 Acta Crystallographica Section E
Structure Reports
Online
ISSN 1600-5368 Acta Crystallographica Section E
Structure Reports
Online
ISSN 1600-5368 Guido J. Reiss* and Sara Bajorat Institut fu¨r Anorganische Chemie und Strukturchemie, Lehrstuhl fu¨r Material- und
Strukturforschung, Heinrich-Heine-Universita¨t Du¨sseldorf, Universita¨tsstrasse 1,
40225 Du¨sseldorf, Germany
Correspondence e-mail: reissg@uni-duesseldorf.de Institut fu¨r Anorganische Chemie und Strukturchemie, Lehrstuhl fu¨r Material- und
Strukturforschung, Heinrich-Heine-Universita¨t Du¨sseldorf, Universita¨tsstrasse 1,
40225 Du¨sseldorf, Germany
Correspondence e-mail: reissg@uni-duesseldorf.de Received 22 November 2007; accepted 30 November 2007 Received 22 November 2007; accepted 30 November 2007 Key indicators: single-crystal X-ray study; T = 293 K; mean (C–C) = 0.002 A˚;
R factor = 0.038; wR factor = 0.072; data-to-parameter ratio = 22.4. Key indicators: single-crystal X-ray study; T = 293 K; mean (C–C) = 0.002 A˚;
R factor = 0.038; wR factor = 0.072; data-to-parameter ratio = 22.4. Table 1
Hydrogen-bond geometry (A˚ , ). D—H A
D—H
H A
D A
D—H A
N1—H3 Cl1ii
0.88 (2)
2.30 (2)
3.1734 (15)
170.3 (16)
N1—H2 Cl1iii
0.86 (2)
2.33 (2)
3.1833 (13)
171.9 (17)
N1—H1 Cl1
0.93 (2)
2.23 (2)
3.1584 (13)
173.9 (16)
Symmetry codes: (ii) x 1
2; y þ 1
2; z 1
2; (iii) x þ 1
2; y þ 1
2; z 1
2. A redetermination of the crystal structure of the title
compound, C6H16N2
2+2Cl, was undertaken. All atomic
coordinates including those of the H atoms were refined
freely. The cation is located on a centre of symmetry. Important for the crystal structure are wavy hydrogen-bonded
layers that are formed by ammonium groups and chloride
anions, giving hydrogen-bonded R3
6ð12Þ rings. Data collection: CrysAlis CCD (Kuma Diffraction, 2000); cell
refinement: CrysAlis RED (Kuma Diffraction, 2000); data reduction:
CrysAlis RED; program(s) used to solve structure: SHELXS97
(Sheldrick, 1997); program(s) used to refine structure: SHELXL97
(Sheldrick, 1997); molecular graphics: DIAMOND (Brandenburg,
2001);
software
used
to
prepare
material
for
publication:
SHELXL97. Related literature For previous structure determinations, see: Dunitz & Strickler
(1965, 1966). For the isostructural cyclohexane-1,4-diammo-
nium dibromide, see: Rademeyer (2006). For hydrogen-bond
motifs, see: Etter et al. (1990); Rademeyer (2006). Supplementary data and figures for this paper are available from the
IUCr electronic archives (Reference: GD2030). Brandenburg, K. (2001). DIAMOND. Crystal Impact GbR, Bonn, Germany.
Dunitz, J. D. & Strickler, P. (1965). Helv. Chim. Acta, 48, 1450–1456.
Dunitz, J. D. & Strickler, P. (1966). Helv. Chim. Acta, 49, 2502–2505.
Etter, M. C., MacDonald, J. C. & Bernstein, J. (1990). Acta Cryst. B46, 256–262.
Kuma Diffraction (2000). CrysAlis CCD and CrysAlis RED. Versions 1.166.
Kuma Diffraction Instruments, Wroclaw, Poland.
Rademeyer, M. (2006). Acta Cryst. E62, o5767–o5769.
Sheldrick, G. M. (1997). SHELXS97 and SHELXL97. University of
Go¨ttingen, Germany. Guido J. Reiss and Sara Bajorat S1. Comment References Brandenburg, K. (2001). DIAMOND. Crystal Impact GbR, Bonn, Germany. Dunitz, J. D. & Strickler, P. (1965). Helv. Chim. Acta, 48, 1450–1456. Dunitz, J. D. & Strickler, P. (1966). Helv. Chim. Acta, 49, 2502–2505. Etter, M. C., MacDonald, J. C. & Bernstein, J. (1990). Acta Cryst. B46, 256–262. Kuma Diffraction (2000). CrysAlis CCD and CrysAlis RED. Versions 1.166. Kuma Diffraction Instruments, Wroclaw, Poland. Rademeyer, M. (2006). Acta Cryst. E62, o5767–o5769. Sheldrick, G. M. (1997). SHELXS97 and SHELXL97. University of
Go¨ttingen, Germany. Brandenburg, K. (2001). DIAMOND. Crystal Impact GbR, Bonn, Germany. Dunitz, J. D. & Strickler, P. (1965). Helv. Chim. Acta, 48, 1450–1456. Dunitz, J. D. & Strickler, P. (1966). Helv. Chim. Acta, 49, 2502–2505. Etter, M. C., MacDonald, J. C. & Bernstein, J. (1990). Acta Cryst. B46, 256–262. Kuma Diffraction (2000). CrysAlis CCD and CrysAlis RED. Versions 1.166. Kuma Diffraction Instruments, Wroclaw, Poland. Rademeyer, M. (2006). Acta Cryst. E62, o5767–o5769. Sheldrick, G. M. (1997). SHELXS97 and SHELXL97. University of
Go¨ttingen, Germany. Acta Cryst. (2008). E64, o223 Reiss and Bajorat
o223 doi:10.1107/S1600536807064793 supporting information Acta Cryst. (2008). E64, o223 [https://doi.org/10.1107/S1600536807064793] Acta Cryst. (2008). E64, o223 [https://doi.org/10.1107/S1600536807064793] Redetermination of trans-cyclohexane-1,4-diammonium dichlorid S1. Comment The title compound was first crystallographically characterized by Dunitz and coworkers in 1966 (Dunitz &
Strickler,1965, 1966). This quality structure determination only lacks the fact that all hydrogen atom positions, especially
those of the ammonium group, were introduced into the structure model on the basic of geometrically calculated
positions, with the N—H and the C—H distances set to 1.1 Å. For X-ray data refinement of hydrogen atom positions
significantly shorter values are commonly found. We now describe an improved structure model - the hydrogen atoms
were reliably found and refined from quality X-ray data. A standard refinement using reflections up to 50 ° / 2Θ gave the following values: R2 = 6.01, R1 = 2.97, GooF = 1.197. Using data with reflections up to 65°/2Θ a more stable refinement is possible and the standard uncertainies of the N—H-
distances are smaller. The title structure consists of hydrogen bonded hydrophilic layers in the ac-plane. These wavy layers are built by an
annulated ring-motif (R36(12); Etter, 1990) constructed by three chloride anions and three ammonium groups (Fig. 3). Each ammonium group donates three hydrogen bonds of only slightly different strength to neighbouring chloride anions
(Tab. 2, Fig. 1 + 2). The title compound is therefore isostructual but not isotypic to the cyclohexane-1,4-diammonium
dibromide (Rademeyer, 2006). In terms of crystal engineering the structure of the title compound is dominated by the hydrogen bonded layers. The
aliphatic cyclohexane-1,4-diyl fragments connect these layers. According to the positions of the ammonium groups in the
hydrogen bonded network the cyclohexyl-fragments do not appear cloesly packed (Fig. 3). trans-Cyclohexane-1,4-diammonium dichloride was prepared by the reaction of 1,4-diaminocyclohexane (+99%, Aldrich,
0.11 g) and hydrochloric acid (37%) at room temperature. From this colourless solution small block shaped crystals were
obtained. trans-Cyclohexane-1,4-diammonium dichloride was prepared by the reaction of 1,4-diaminocyclohexane (+99%, Aldrich,
0.11 g) and hydrochloric acid (37%) at room temperature. From this colourless solution small block shaped crystals were
obtained. S3. Refinement S3. Refinement All hydrogen atom positions were obtained from difference fourier maps, all hydrogen atoms were refined freely and
with an individual isotropic displacement parameter for each (H—X distance range: 0.88–1.01 A). All hydrogen atom positions were obtained from difference fourier maps, all hydrogen atoms were refined freely and
with an individual isotropic displacement parameter for each (H—X distance range: 0.88–1.01 A). sup-1 Acta Cryst. (2008). E64, o223 supporting information supporting information supporting information
Figure 1
The structure of the title compound (displacement ellipsoids at the 40% probability level, H-atoms drawn with arbitrary
radius).Thin dashed lines show hydrogen bonds to neighbouring chloride anions. The atoms of the asymmetric unit are
labeled. Figure 1
The structure of the title compound (displacement ellipsoids at the 40% probability level, H-atoms drawn with arbitrary
radius).Thin dashed lines show hydrogen bonds to neighbouring chloride anions. The atoms of the asymmetric unit are
labeled. g
The structure of the title compound (displacement ellipsoids at the 40% probability level, H-atoms drawn with arbitrary
radius).Thin dashed lines show hydrogen bonds to neighbouring chloride anions. The atoms of the asymmetric unit are
labeled. The structure of the title compound (displacement ellipsoids at the 40% probability level, H-atoms drawn with arbitrary
radius).Thin dashed lines show hydrogen bonds to neighbouring chloride anions. The atoms of the asymmetric unit are
labeled. sup-2 sup-2 Acta Cryst. (2008). E64, o223 supporting information pp
g
Figure 2 supporting information supporting information Data collection
Stoe STADI CCD
diffractometer
Radiation source: fine-focus sealed tube
Graphite monochromator
ω scans
13502 measured reflections
1766 independent reflections
Refinement
Refinement on F2
Least-squares matrix: full
R[F2 > 2σ(F2)] = 0.038
wR(F2) = 0.072
S = 1.04
1766 reflections
79 parameters
0 restraints
Primary atom site location: structure-invariant
direct methods Data collection
Stoe STADI CCD
diffractometer
Radiation source: fine-focus sealed tube
Graphite monochromator
ω scans
13502 measured reflections
1766 independent reflections
Refinement
Refinement on F2
Least-squares matrix: full
R[F2 > 2σ(F2)] = 0.038
wR(F2) = 0.072
S = 1.04
1766 reflections
79 parameters
0 restraints
Primary atom site location: structure-invariant
direct methods Data collection
Stoe STADI CCD
diffractometer
Radiation source: fine-focus sealed tube
Graphite monochromator
ω scans
13502 measured reflections
1766 independent reflections
Refinement
Refinement on F2
Least-squares matrix: full
R[F2 > 2σ(F2)] = 0.038
wR(F2) = 0.072
S = 1.04
1766 reflections
79 parameters
0 restraints
Primary atom site location: structure-invariant
direct methods 1562 reflections with I > 2σ(I)
Rint = 0.043
θmax = 32.5°, θmin = 4.3°
h = −7→7
k = −22→22
l = −8→9 Stoe STADI CCD
diffractometer
Radiation source: fine-focus sealed tube
Graphite monochromator
ω scans
13502 measured reflections
1766 independent reflections Refinement
Refinement on F2
Least-squares matrix: full
R[F2 > 2σ(F2)] = 0.038
wR(F2) = 0.072
S = 1.04
1766 reflections
79 parameters
0 restraints
Primary atom site location: structure-invariant
direct methods Secondary atom site location: difference Fourier
map
Hydrogen site location: difference Fourier map
All H-atom parameters refined
w = 1/[σ2(Fo2) + 0.3P]
where P = (Fo2 + 2Fc2)/3
(Δ/σ)max < 0.001
Δρmax = 0.41 e Å−3
Δρmin = −0.28 e Å−3
Extinction correction: SHELXL,
Fc*=kFc[1+0.001xFc2λ3/sin(2θ)]-1/4
Extinction coefficient: 0.174 (6) Secondary atom site location: difference Fourier
map
Hydrogen site location: difference Fourier map
All H-atom parameters refined
w = 1/[σ2(Fo2) + 0.3P]
where P = (Fo2 + 2Fc2)/3
(Δ/σ)max < 0.001
Δρmax = 0.41 e Å−3
Δρmin = −0.28 e Å−3
Extinction correction: SHELXL,
Fc*=kFc[1+0.001xFc2λ3/sin(2θ)]-1/4
Extinction coefficient: 0.174 (6) Hydrogen site location: difference Fourier map All H-atom parameters refined Figure 2 Figure 2
Hydrogen bonded layer in the ac-plane of the title structure, constructed from annulated R36(12) motifs. (Symmetry code ′
= -1/2 + x, 0,5 - y, -1/2 + z) Hydrogen bonded layer in the ac-plane of the title structure, constructed from annulated R36(12) motifs. (Sym
= -1/2 + x, 0,5 - y, -1/2 + z)
Figure 3
Crystal packing seen along the c direction. Hydrogen bonding interactions are shown as dotted lines. Hydrogen bonded layer in the ac-plane of the title structure, constructed from annulated R 6(12) motifs. (S
= -1/2 + x, 0,5 - y, -1/2 + z)
Figure 3 g
Crystal packing seen along the c direction. Hydrogen bonding interactions are shown as dotted lines. g
Crystal packing seen along the c direction. Hydrogen bonding interactions are shown as dotted lines. trans-cyclohexane-1,4-diammonium dichloride trans-cyclohexane-1,4-diammonium dichloride Crystal data
C6H16N22+·2Cl−
Mr = 187.11
Monoclinic, P21/n
a = 5.2550 (11) Å
b = 14.890 (3) Å
c = 6.3604 (12) Å
β = 99.824 (18)°
V = 490.39 (16) Å3
Z = 2
F(000) = 200
Dx = 1.267 Mg m−3
Mo Kα radiation, λ = 0.71073 Å
Cell parameters from 1610 reflections
θ = 4.8–17.4°
µ = 0.60 mm−1
T = 293 K
Block, colourless
0.30 × 0.24 × 0.20 mm Crystal data
C6H16N22+·2Cl−
Mr = 187.11
Monoclinic, P21/n
a = 5.2550 (11) Å
b = 14.890 (3) Å
c = 6.3604 (12) Å
β = 99.824 (18)°
V = 490.39 (16) Å3
Z = 2
F(000) = 200
Dx = 1.267 Mg m−3
Mo Kα radiation, λ = 0.71073 Å
Cell parameters from 1610 reflections
θ = 4.8–17.4°
µ = 0.60 mm−1
T = 293 K
Block, colourless
0.30 × 0.24 × 0.20 mm F(000) = 200
Dx = 1.267 Mg m−3
Mo Kα radiation, λ = 0.71073 Å
Cell parameters from 1610 reflections
θ = 4.8–17.4°
µ = 0.60 mm−1
T = 293 K
Block, colourless
0.30 × 0.24 × 0.20 mm sup-3 sup-3 Acta Cryst. (2008). E64, o223 Special details Geometry. All e.s.d.'s (except the e.s.d. in the dihedral angle between two l.s. planes) are estimated using the full
covariance matrix. The cell e.s.d.'s are taken into account individually in the estimation of e.s.d.'s in distances, angles and
torsion angles; correlations between e.s.d.'s in cell parameters are only used when they are defined by crystal symmetry. An approximate (isotropic) treatment of cell e.s.d.'s is used for estimating e.s.d.'s involving l.s. planes. Refinement. Refinement of F2 against ALL reflections. The weighted R-factor wR and goodness of fit S are based on F2,
conventional R-factors R are based on F, with F set to zero for negative F2. The threshold expression of F2 > σ(F2) is used
only for calculating R-factors(gt) etc. and is not relevant to the choice of reflections for refinement. R-factors based on F2
are statistically about twice as large as those based on F, and R- factors based on ALL data will be even larger. Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Å2)
x
y
z
Uiso*/Ueq
Cl1
0.46825 (6)
0.15413 (2)
0.46296 (5)
0.03915 (12)
N1
0.5260 (2)
0.34260 (8)
0.24820 (19)
0.0338 (2)
H1
0.522 (3)
0.2870 (13)
0.316 (3)
0.051 (5)*
H2
0.642 (4)
0.3377 (12)
0.168 (3)
0.057 (5)*
H3
0.377 (4)
0.3498 (12)
0.163 (3)
0.050 (5)*
C1
0.6023 (3)
0.50463 (8)
0.3001 (2)
0.0335 (3)
H11
0.448 (3)
0.5150 (11)
0.198 (3)
0.041 (4)*
H12
0.738 (4)
0.5022 (11)
0.221 (3)
0.043 (4)*
C2
0.5751 (2)
0.41590 (8)
0.41008 (18)
0.0281 (2)
H21
0.726 (3)
0.4018 (10)
0.503 (2)
0.029 (3)*
C3
0.3565 (3)
0.41864 (9)
0.5386 (2)
0.0358 (3)
H31
0.203 (3)
0.4276 (11)
0.445 (3)
0.042 (4)*
H32
0.355 (3)
0.3590 (11)
0.613 (3)
0.044 (4)* actional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Å2) Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Å2)
x
y
z
Uiso*/Ueq sup-4 Acta Cryst. (2008). Symmetry code: (i) −x+1, −y+1, −z+1. Special details E64, o223 pp
g
Atomic displacement parameters (Å2)
U11
U22
U33
U12
U13
U23
Cl1
0.03850 (18)
0.03987 (18)
0.04168 (19)
−0.00250 (12)
0.01427 (13)
0.00190 (13)
N1
0.0361 (5)
0.0330 (5)
0.0350 (5)
−0.0006 (4)
0.0137 (4)
−0.0018 (4)
C1
0.0416 (6)
0.0331 (6)
0.0301 (6)
−0.0022 (5)
0.0179 (5)
0.0026 (4)
C2
0.0275 (5)
0.0315 (5)
0.0267 (5)
0.0006 (4)
0.0082 (4)
0.0021 (4)
C3
0.0394 (6)
0.0341 (6)
0.0390 (6)
−0.0078 (5)
0.0211 (5)
−0.0008 (5)
Geometric parameters (Å, º)
N1—C2
1.4924 (16)
C2—C3
1.5201 (16)
C1—C2
1.5130 (17)
C2—H21
0.928 (15)
C1—C3i
1.5259 (18)
C3—C1i
1.5259 (18)
C1—H11
0.959 (17)
C3—H31
0.927 (17)
C1—H12
0.941 (19)
C3—H32
1.008 (17)
C2—N1—H1
110.2 (11)
N1—C2—C3
109.47 (10)
C2—N1—H2
114.4 (13)
N1—C2—C1
109.79 (10)
H1—N1—H2
105.8 (16)
C3—C2—C1
111.39 (10)
C2—N1—H3
111.6 (12)
N1—C2—H21
107.7 (9)
H1—N1—H3
107.8 (16)
C3—C2—H21
107.6 (9)
H2—N1—H3
106.8 (18)
C1—C2—H21
110.8 (9)
C2—C1—C3i
110.91 (10)
C2—C3—C1i
110.36 (10)
C2—C1—H11
108.6 (10)
C2—C3—H31
108.1 (11)
C3i—C1—H11
110.0 (10)
C1i—C3—H31
109.5 (10)
C2—C1—H12
110.8 (10)
C2—C3—H32
107.4 (10)
C3i—C1—H12
110.9 (10)
C1i—C3—H32
110.8 (10)
H11—C1—H12
105.6 (15)
H31—C3—H32
110.6 (15)
C3i—C1—C2—N1
178.07 (10)
N1—C2—C3—C1i
−177.94 (11)
C3i—C1—C2—C3
56.65 (16)
C1—C2—C3—C1i
−56.34 (16) Atomic displacement parameters (Å2) Geometric parameters (Å, º) Hydrogen-bond geometry (Å, º)
D—H···A
D—H
H···A
D···A
D—H···A
N1—H3···Cl1ii
0.88 (2)
2.30 (2)
3.1734 (15)
170.3 (16)
N1—H2···Cl1iii
0.86 (2)
2.33 (2)
3.1833 (13)
171.9 (17)
N1—H1···Cl1
0.93 (2)
2.23 (2)
3.1584 (13)
173.9 (16)
Symmetry codes: (ii) x−1/2, −y+1/2, z−1/2; (iii) x+1/2, −y+1/2, z−1/2. Hydrogen-bond geometry (Å, º) sup-5 Acta Cryst. (2008). E64, o223 sup-5
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Leadership as an Act of Love: Leading in Dangerous Times
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Mónica Byrne-Jiménez
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Leadership as an Act of Love:
Leading in Dangerous Times Mónica C. Byrne-Jiménez 1* and Irene H. Yoon 2 1 Educational Leadership and Policy Studies, Indiana University, Bloomington, IN, United States, 2 Educational Leadership
and Policy, University of Utah, Salt Lake City, UT, United States 1 Educational Leadership and Policy Studies, Indiana University, Bloomington, IN, United States, 2 Educational Leadership
and Policy, University of Utah, Salt Lake City, UT, United States This essay is an invitation to engage in a heart-whole thought experiment around the
possibilities and potential of framing leadership as an act of love. The current school
context is defined by increasing pressures and demands that undermines the humanity
of leaders, children and educators. In this context, leaders must find other ways to lead. The authors propose a framework offered to help leaders to reconnect with their love
of children and education. They define the “habits of the heart” as harmony, courage,
wisdom, and imagination. It is the combination of these habits that allow leadership with
and through love. By defining these habits of heart, leaders can recognize them in their
existing practice and find ways to enact new practices that create schools of learning
and belonging. Keywords: leadership, love, schools, heart, humanity, preparation Those of us who have already chosen to embrace a love ethic, allowing it to govern and inform how we
think and act, know that when we let our light shine, we draw to us and are drawn to other bearers of light. We are not alone. Edited by:
Lauri Johnson,
Boston College, United States Edited by:
Lauri Johnson,
Boston College, United States (Hooks, 2001,101) CONCEPTUAL ANALYSIS
published: 09 January 2019
doi: 10.3389/feduc.2018.00117 Edited by:
Lauri Johnson,
Boston College, United States
Reviewed by:
Christa Boske,
Kent State University, United States
Martin Scanlan,
Boston College, United States
*Correspondence:
Mónica C. Byrne-Jiménez
mcbyrnej@iu.edu PREFACE Reviewed by:
Christa Boske,
Kent State University, United States
Martin Scanlan,
Boston College, United States These are complicated and complex times. Globally, populist waves are sweeping democracy
aside and putting authoritarian rulers in its stead, the most recent being the presidential election
in Brazil. Within the United Sates, the rise of White Supremacy and its acceptance into the
halls—and coffers—of government is but only one example. In both contexts, we see increasing
disrespect for and aggressive assertions of power over marginalized communities, along with the
rise of neo-liberal economic and social policies that value profit over people. If public institutions,
educational and otherwise, are functioning in a state of persistent crisis, then we must rethink the
capacity of leaders and schools to stem this tide of fear and hate. What does it mean to reject these
impulses and emotions and to embrace all members of a community? We argue that these times
require a unique form of leadership that draws its strength—not from policies or regulations or
authority, but from the heart in communion with others. *Correspondence:
Mónica C. Byrne-Jiménez
mcbyrnej@iu.edu Specialty section:
This article was submitted to
Leadership in Education,
a section of the journal
Frontiers in Education
Received: 07 August 2018
Accepted: 13 December 2018
Published: 09 January 2019
Citation:
Byrne-Jiménez MC and Yoon IH
(2019) Leadership as an Act of Love:
Leading in Dangerous Times. Front. Educ. 3:117. doi: 10.3389/feduc.2018.00117 Specialty section:
This article was submitted to
Leadership in Education,
a section of the journal
Frontiers in Education There has been recent and increasing attention in educational discourse on mindfulness (Yaron,
2015), the role of empathy in learning (see the recent special issue on “Learning with Empathy”
in ASCD Express, September 2017), and social-emotional learning (e.g., Durlak et al., 2011). We support those conversations whole-heartedly (pun intended). We also believe that, for those
conversations to have an impact on leader practice and leadership preparation, these ideas must
be defined and integrated into a “conceptual framework.” This task must be deliberative and taken
up with care. We understand that the framework we offer here would need to be tested, improved,
and expanded. We offer it in order to foster debate and more critical thinking of the relationship
between leadership and love. It is the starting point for further discussion and study. OUR PURPOSE We exist in the in-between,
often negotiating personal and professional identities out loud
and in public spaces, like this one. What brings us together
are not only our stories and the layers within them, but also
how we tell them–with laughter and optimism, and a heartfelt
recognition of both loss and pain. Our stories are inseparable
from how we have experienced this current moment in history
and the school leadership that it calls for. One major issue that has resurfaced in the past decade is
the need for social-emotional learning and trauma-informed
care. Duncan-Andrade and Morrell (2008) have written of
children who suffer from persistent (as opposed to post-)
traumatic stress disorder. These are students who have significant
social, emotional, and/or intellectual challenges because of
long term, ongoing
exposure to poverty, institutionalized
racism, residential instability, immigration status, hunger,
violence, and criminalization by court systems. These complex
traumas are historical with transgenerational effects on students’
neurological, social, and emotional development. Along with
greater recognition of trauma and its long term negative impact
on children’s well-being is increasing acknowledgment that
schools often retraumatize students with punitive discipline, lack
of responsiveness, and disrespect (Shaia and Crowder, 2017). Despite the best efforts of some educators, schools often add
further stress to students’ lives. Students are resilient, but these
are perilous times for children, for adults ill-equipped for the
level of support children need, and for school leaders who
must address holistic needs while improving achievement on
standardized tests. In addition, leaders and staffmay experience
burnout and secondary traumatization [or “compassion fatigue,”
(Conrad and Kellar-Guenther, 2006)] from their own repeated
exposure to students’ or community trauma (Alisic, 2012). These
issues are not new, but rather have been illuminated from
recent dissemination of research in mental health, social work,
counseling, sociology, cultural studies, and history. Though this
literature has existed for some time, teacher and leadership
preparation programs have not attended to the mental health
needs of the children and of the adults who attempt to support
them in chronically under-resourced systems. These are indeed
dangerous times. The invitation to engage in this thought experiment, therefore,
is by its very nature an integration of the academic and the
personal, the mind and the heart. As scholars we are intellectual
and emotional human beings. OUR PURPOSE In schools around the world, children arrive every day with
a host of hopes and needs. Teachers arrive with their own
hopes and needs. School leaders also arrive with their hopes
and needs. For adults, these needs are personal as much as
professional. These needs, for children and adults, are often
ignored during the course of the school day and, perhaps,
even the school year. Schools, and the people within them, are
governed by rules regarding curriculum (scope and sequences,
assessments), roles and responsibilities (in- vs. out-of-classroom
personnel, students, parents), structures (schedules, committees),
and systems (communication, discipline). These rules are,
more often than not, inflexible and independent of those
who implement them or those they serve. Leaders are tasked
with making sure that these structures and processes operate
efficiently and effectively, yet they are also keenly aware of the
adults and children in their care. In schools around the world, children arrive every day with
a host of hopes and needs. Teachers arrive with their own
hopes and needs. School leaders also arrive with their hopes
and needs. For adults, these needs are personal as much as
professional. These needs, for children and adults, are often
ignored during the course of the school day and, perhaps,
even the school year. Schools, and the people within them, are
governed by rules regarding curriculum (scope and sequences,
assessments), roles and responsibilities (in- vs. out-of-classroom
personnel, students, parents), structures (schedules, committees),
and systems (communication, discipline). These rules are,
more often than not, inflexible and independent of those
who implement them or those they serve. Leaders are tasked
with making sure that these structures and processes operate
efficiently and effectively, yet they are also keenly aware of the
adults and children in their care. We acknowledge and affirm that our views come from our
specific journeys and identities. We are both cisgender women
of color, the daughters of immigrants, from bilingual homes
and family histories that continually cross borders. Mónica is
a daughter of Mexican and Peruvian parents. Irene is Korean
American, with a “hidden” disability. Both of us attended elite
colleges and have spent our careers working for educational
equity. Both of us work in organizations that often challenge how
we move in the world and force us to make tiny, daily adjustments
in order to thrive in those spaces. Citation: January 2019 | Volume 3 | Article 117 Frontiers in Education | www.frontiersin.org Byrne-Jiménez and Yoon Leading With Love even before this essay reaches you. In light of that tragedy, the need
for re-visioning or re-imagining school leadership becomes even
clearer. We dedicate this essay to the families who have lost a loved
one through school violence. In
November
2016,
Mónica
Byrne-Jiménez
gave
her
Presidential Address at the UCEA Annual Convention (Byrne-
Jiménez, 2017). In that speech Dr. Byrne-Jiménez challenged
faculty in educational leadership to reclaim love as an “active
and important part of [their] leadership” (2). She spoke from a
place where her intellectual vision for the field was inseparable
from her heart—her belief in education, her values, and her
commitments to justice. Subsequently, Irene Yoon wrote a
response for the AERA Division A Newsletter (Yoon, 2017). Dr. Yoon added her thoughts to what “tough-minded, tender-
hearted” leadership with love would look like. This essay builds
on both of those to deepen the discussion, transform it into a
dialogue, and urge the field of educational leadership to develop
ways of leading that are more humanizing and responsive to the
demands of growing up in the twenty-first century. As educators,
and those who prepare future leaders, our job ultimately is to
create schools that are places of joy, shelter, and learning. Frontiers in Education | www.frontiersin.org OUR PURPOSE speed by which we transmit cultural norms in discrete, easily
digestible “memes,” the fact that Facebook has created a new unit
of time called a “Flick,” which is one “five million six hundred
thousandth of a second” (Coldewey, 2018, emphasis added),
all highlight how nothing can stand in the way of time. In a
world where time is compressed and information—real or not—
is shared in a blink of an eye, there is often little opportunity to
explore the meaning of that information or how it might affect
our existence. How we measure time is not the only challenge. Because
of social media and the 24-h news cycle we can know more
about the world than ever before. And because of—or perhaps
despite—the global economy we are increasingly aware of the
human connections that exist around the globe. It is difficult to
remain unmoved when we see the plight and determination of
Syrian refugees or the devastation of floods in Bangladesh or the
fear in the face of slaves mining for precious metals in Africa
or the horror in the face of children who survived an attack
on their school. Recent environmental movements and research
have noted that the environmental disasters of climate change
are occurring more rapidly than earlier predictions, and have
stressed the belief that we are as connected to each other as we
are to the Earth. Damage done to one part of the Earth affects us
all. In this essay, we offer our thoughts on the current educational
context and the new and unique pressures it is putting on
educational leaders, school principals in particular. We take
note of the confluence of forces that are focused on public
education broadly and the implications for preparing leaders
who can be joyful even as they engage in the hard work of
educating children and preparing them with the skills and
heart to be resilient, loving, community members. In order to
enter this creative space—or thought experiment—we propose
a leadership framework that re-centers elements of liberatory
education (Freire, 1970; Darder, 1998), tempered radicalism
(Meyerson and Scully, 1995), and loving epistemology (Liable,
2000; see also Capper, 2000). These scholars remind us that for
too long organizations—schools in particular—have forced us
to decide between our work and our humanity. Organizations
employ several strategies to force a wedge not just within each of
us, but among all of us. OUR PURPOSE This distance, this othering of ourselves
(Capper, 2000) from ourselves and from each other—or what
Liable (2000) calls “institutional evil”—serves to further isolate
us from the very thing that would make us stronger: recognition
of each other’s humanity. In contrast, these scholars urge us to
resist those dissociative forces and find ways to maintain our
integrity as inter- and intra-connected human beings. The work
of leadership is messy and requires that we enter the murky
world of relationships. At the core of our thinking—and feeling—
we know that leadership does matter to children, schools, and
communities around the country, indeed around the world. We
must bring the full weight of our profession, our skills, our energy
to bear on reclaiming the heart of leadership. Thus, we imagine a
leadership grounded in radical, transformational, and sustained
love. We also exist in a more polarized political world. Rising
nationalist and fascist movements endanger us all, especially
the most marginalized who are the targets of this dangerous
rhetoric. The need to work together for the common good is
more important than ever. We work together—we care for each
other—or we go down together. g
g
If nothing else, the above clearly indicate that leadership can
no longer be about the search for a “theory of everything” or
a set of static “best practices.” It is about envisioning multiple
ways forward in order to prepare schools, teachers, and students
for something they cannot see or predict. To do this, we
must prepare leaders who are constantly adapting and thinking,
planning for the future, yet who do not lose their footing or
the “fierce urgency of now” (King, 1967). Crisis and uncertainty
are also moments of great opportunity for social movements,
democratic engagement, and collective action if we have the
will to do the work that’s needed. Leaders at all levels must
be taught to reach across boundaries that divide us, that have
been constructed and that we—all of us—continue to enable,
consciously or not. In these contexts, leaders who embrace
uncertainty and who understand that the journey has several
unknown destinations will be able to prepare students for a
myriad of possibilities. OUR PURPOSE We ask you, then, to suspend
judgments or expectations of traditional scholarly dialogue and
join us in probing the limits of our collective understanding of
leadership. By doing so, we begin to exercise new skills within
the framework we propose and learn to act, think, and lead
in new ways. This is not a typical way of dialoguing with our
peers, and yet it honors our scholarly interests and reflects a
commitment to finding ways of being that center our common
humanity. Relatedly, we are scholars grounded in the work of
schools and the lived experiences of practitioners. We see our
work as connected to schools and classrooms and understand the
tensions that emerge when we try to do this while simultaneously
responding to the demands of our daily work in institutions of
higher education. We hold that tension as part of our work and
invite you to join us as we explore what a form of leadership that
leads with love could look like. Since we began writing this, there have been two more school
shootings in the United States. Without action from legislators and
political leaders, we mourn in the knowledge that more will occur Leadership preparation programs have long reframed the
role of school leadership from organizational management to January 2019 | Volume 3 | Article 117 Frontiers in Education | www.frontiersin.org 2 Leading With Love Byrne-Jiménez and Yoon instructional leadership (e.g., Poplin, 1992; Elmore and Burney,
1997). More recent efforts have emphasized principals as leaders
for social justice and equity, focusing on addressing gaps in
opportunity, achievement, and life outcomes that result from
structural deficits (e.g., Dantley and Tillman, 2010). Modern
leadership requires a new way of addressing the needs of children
and adults. This is not to say that leaders need to do more, but
rather that they need to do better. One element of that is to
reclaim love as both a leadership strategy and as a leadership
framework. Leaders must tap into the deep well of their love
for children, for their work, and a deep belief in the power of
education. To prepare this leader, preparation programs must
also recover their capacity to love and their belief that leadership
matters. January 2019 | Volume 3 | Article 117 Frontiers in Education | www.frontiersin.org HABITS OF THE HEART In order to reach a place of love, we must embrace
our anger and use it to strengthen the heart for the long journey
toward justice. j
We, too, believe that love is an energy that emanates from
within to shield and care for others. And since it comes from
within, we must respect its power and be careful not to misuse
it. Love, therefore, both takes risks and provides safety and sense
of belonging. It motivates us to dream beyond our current limits
and grow in body, mind, and spirit. It is a mutually accepted
challenge that is reciprocal at its core and that requires kindness
to oneself and others. Love is wise and wizened from the passage
of time, yet joyful and flexible because it takes delight in the
wonder of youth. Love experiences pain alongside others and
heals through grace. It grows strong because it accepts that
discomfort leads to new understandings. Love is impatient in the
face of injustice while patient in its ability to forgive. Love allows
us to resist hate. p
p
Palmer (1983) brought the minds, hearts, and souls of the
learner and the learned together in his search of an educational
system that cultivated wisdom and relationships. With a similar
goal, we offer four habits of the heart that, when situated in the
work of schools, become guiding principles of forms of leading. We describe leadership as love in terms of habits because they
are ways of being that, with time, become accepted leadership
practices. These habits, which surround and sustain leaders,
form connections between vision and management, external
pressures and internal conflicts, and between young people and
adults, transforming the way we act and interact in our teaching,
learning, and becoming. Like love, these habits of leadership are
varied and unique and beautiful. And, like love, they require
practice and attention. Nee-Benham et al. (1998) described leaders’ commitment to
the “public process of loving, of behaving in a loving manner”
(145). Love, often relegated to the private domain, comes to light
in the leadership and relationships of schooling—with students,
teachers, and families. It is the public nature of love and loving
that elevates it as a way of leading and a purpose for leadership. It is powerful because it integrates the personal and the public in
service to children. HABITS OF THE HEART Love as an active form of leadership may seem to challenge
the current state of schooling. School leaders have less time
to develop thoughtful responses or may choose to respond
in ways that privilege efficiency, effectiveness, and the cult of
personality. The constant pull of external forces—policymakers,
district superintendents, parents who know how to assert their
privilege, business interests, to name a few—along with within-
school pressures and complications of bringing their school
vision to life—puts school leaders under constant strain. As
former teachers themselves, most leaders began their careers
from a place of wholeness or, rather, where confidence and joy
defined their work. Becoming a principal often is alienated from
that place of joy and the current context works to undermine
their confidence. Leaders, therefore, move through their days in
a state of stress, balancing what the system says they must do vs. what their hearts tell them needs to be done. It follows, then, that
leaders can become fragmented and without significant support
are in danger of losing a vital piece of themselves. How then, do
we return leaders to a place of joy in their work and maintain
their personal and professional integrity? Thinking about this
question in the context of the demands of school leadership,
Hooks (2009) offers the development of habits of the heart. The
skills and knowledges and values that make one “heartwhole”
(217) are the foundation of a culture, a community, that shares a
“language of healing, of hope, a language of dreams, a language of
belonging” (223). In this section, we elaborate habits of the heart
that constitute love as an active practice of leadership. In his book, Strength to Love (1963), Martin Luther King Jr. explores the depth and breadth of what he calls divine love. The title alone indicates that love that is sacred because it
connects all living things, is not for the faint-hearted. Indeed, it is
important to also acknowledge the connection between “armed
love” (Freire, 1998) and the bright, clear anger at the core. The
anger comes from witnessing oppression and injustice meted out
on the most vulnerable. This courageous love we are speaking
of is not despite the anger, it is beyond it. Keating (2013), in
a similar spirit, developed a theory of post-oppositionality that
moves beyond the us/them and either/or world view and seeks
rather to adopt relational, transformative, and visionary ways of
interacting. NATURE OF LOVE respect, and responsibility (Hooks, 1994, 2001). Leadership as
love means choosing to be truly present with children and adults
in the building, and creating emotional and intellectual spaces to
which children and adults can return each day (Kessler, 2000). It
is not simply a disposition or feeling; it is an active practice that
transforms schools into spaces of learning and belonging. The capacity to love and to be loved may be one of the
greatest gifts of humankind. It may also be one of our greatest
challenges. Even to write in a scholarly journal about “love”
makes some uncomfortable. In her speech, Byrne-Jiménez drew
a distinction between the “love that you see in the movies”
and a “courageous love that requires everything.” This concept
of an active, courageous love is grounded in a long line of
critical writings from Thomas Merton to Paulo Freire to Gloria
Anzaldúa. These writers, in many ways, see love as “rooted in
a committed willingness to struggle persistently with purpose in
our life and to intimately connect that purpose with what [Friere]
called our “true vocation”—to be human” (Darder, 1998, 498). The search for love, therefore, is a search for our humanity. LEADERSHIP IN A COMPLEX WORLD In the following exploration, we center school principals as the
specific perspective of school leadership for which we envision
the nature of love and the habits of leading with love. Though
we recognize that educational leaders are also in district offices,
higher education institutions, classrooms, school boards, and
state agencies; we adopt the K-12 school principal’s perspective
as an initial foray. We believe that the particular habits of love
are likely to take different forms depending on the position and
context of each leadership role, but similar values and feelings will
likely hold true for all. We are not the first to call for preparing leaders for different,
dynamic contexts (Kay and Greenhill, 2013). We do believe
that new understandings from research and changing political
contexts call for a constant re-examination of core values
and principles of practice. Current leadership frameworks have
evolved, slowly, over time. The world, however, moves at
lightning speeds. The speed of high-frequency trading on the
stock market, the speed by which something goes “viral,” the January 2019 | Volume 3 | Article 117 Frontiers in Education | www.frontiersin.org 3 Leading With Love Byrne-Jiménez and Yoon Frontiers in Education | www.frontiersin.org January 2019 | Volume 3 | Article 117 HABITS OF THE HEART One need only walk through an elementary
school and see children hug their teachers, or in the hallways
of a high school and see the smiles of students as they enter a
classroom, to know that love can live in schools. School leaders
are gardeners who allow love to flourish in their buildings. Leadership in this sense encompasses a love characterized by care, These habits are grounded in what we have described
previously as the nature of love. We draw from the work of many
authors (some cited here), our experiences in and with public
schools, and our identities as female scholars of color. We have
filtered what we learned from these to derive this framework of
four habits of the heart or the ways in which leadership with January 2019 | Volume 3 | Article 117 4 Byrne-Jiménez and Yoon Leading With Love And in doing so we are able to see and embrace the humanity of
others. It also implies that harmony is never truly achieved but
something that we must envision and tend daily. FIGURE 1 | Leadership habits of the heart. We often talk about “balance,” and “finding balance” as if
that were the answer to much of the chaos we experience. Balance implies an equilibrium between two things, weight and
counterweight. But our lives are rarely so bifurcated, our choices
and their consequences rarely so clearly delineated. We have
multiple demands, responsibilities, roles, goals, identities, desires. Rather than balance, we need to seek harmony and to live in ways
that bring harmony within ourselves (our past, our dreams, our
doubts, our gifts), with others, and with the environment. Much of the challenges that leaders face stem from
organizational disharmony (Overvold, 1987), in which the
uniqueness of each person is subsumed to the needs of the
organization. To lead with/in harmony, educational leaders have
to integrate their different selves in to a more whole existence. To lead in harmony also means that educational leaders have
to integrate the different selves in the organization to create a
new and stronger whole. Because leaders constantly negotiate
among the needs of the organization and those of the people
that make it up, they run the risk of creating disharmony
in both. HABITS OF THE HEART This is especially challenging when multiple needs,
interests, roles (i.e., leader, teacher, partner, parent) and identities
(i.e., race/ethnicity, sexual orientation, multilingual/cultural,
ability)—their own and those of others—are clamoring for
attention. Leading and learning, therefore, are physical, affective,
and intellectual experiences woven together to create a new
organizational tapestry. In the midst of this complexity, leaders
must learn to exist in harmony with the communities they lead. FIGURE 1 | Leadership habits of the heart. love is enacted. We named these harmony, courage, wisdom, and
imagination. In our minds these are like the gears of a watch:
individual pieces that work in unison, each invaluable to the
others (see Figure 1). In the next section we will attempt to define
these habits and begin to identify how these might emerge in
principals’ work. In order to make this thought experiment successful, we
encourage you to read with an open heart and to imagine what
these habits could look like in your own practice. What would
your own life be like if you, and the leaders around you, all led
with love? We challenge ourselves and invite our colleagues to engage in
habits of harmony. If we were to imagine habits that indicated
leadership that embraces harmony, we could envision principals
who: Harmony • Listen to their hearts • Welcome their elders and youth When I was a child, someone at school told me that you could melt
a slug by putting salt on it. Skeptical of this piece of information, I
went home, got the salt shaker from the kitchen, and went in search
of a slug. When I came across one, I promptly sprinkled it with salt
and watched it dissolve before my eyes. I cried at my handiwork—
at the violence I had committed—and vowed never to harm a slug
again. To this day, I feel a deep sense of shame of having willfully
killed that slug. I did not have the language or understanding to
realize that, through my selfish act, I had upset the harmony of that
garden, of that day. • Stay present in their community • Invite others to connect and embraces the unknown • Find laughter and joy in the small, everyday even • Give expression to their creative selves Harmony is a journey, and in a world obsessed with destinations,
it can be easy to forget how to stay attuned to yourself, those
around you, nature. And yet it can be the one constant that makes
for complete and heartwhole leaders. Frontiers in Education | www.frontiersin.org January 2019 | Volume 3 | Article 117 Courage can prepare for the worst and not lead their communities to
live in fear. Furthermore, leaders are challenged to confront
casual or implicit racism, able-ism, genderism, or other forms of
exclusion when doing so can cause conflict. Leaders even may
have to make staffing decisions that are unpopular with faculty
but are important for students. Below these questions, leaders
may encounter whispers of self-doubt and fear. It is often said that courage is not the absence of fear,
but rather overcoming it. Leaders’ courage to persevere comes
from an unwavering commitment to a vision of schooling
as an experience of collective growth, passion, and mutual
care among adults, children, and community members. A
community of courage is fundamental for leaders to create an
environment of physical and emotional safety and to sustain
school-wide commitment to a vision of equity and justice in
the face of funding cuts, high turnover or reductions in staffing,
bureaucratic processes that seem unfathomable, and inevitable
disappointments when struggling to support all students well. Withstanding these storms is a daily habit of courageous
love. Because love thrives through connection, courage is
made possible when leaders experience collective joy or tired
laughter, through shared good days and through the excitement
of moments seem magical—when schools and classrooms are
responsive, creative, and playful centers of learning for adults and
students. These moments fuel leaders’ abilities to stay courageous
on behalf of the heart of children and adults in their buildings. Seeing the difference between knowing and wisdom requires a
level of humility not valued in modern society. In a world that
values certainty, control, fast solutions, winners-losers, having
humility can be seen as weakness. What is often misunderstood
is that true strength comes from being able to say “I don’t
know,” or “I was wrong,” or admitting that there are things
to be learned from others different from yourself. Equally
important, the humility to see the wisdom of those who have gone
ahead or are following in our footsteps and/or caring steps of
their own. Courageous love does not settle for mediocrity. Perhaps this
is why we believe that leadership with love needs emotional and
cognitive space to dream for more for their students and staff. When leadership is courageous for audacious and shared dreams,
leaders can navigate and channel emotions—even fear—as tools
to support, engage, empower others. Courage Educational leaders may rely on their prior success (they
would not be school leaders if they had not already been
successful somehow) and training as a measure of their
qualifications. To be sure, having a depth of knowledge of
leadership theory, practices, systems is a necessary, but not
sufficient, part of leadership. Wisdom for leadership comes from
being in the world, present in nature, with elders and children,
and seeking clarity about your role in these realms. It requires a
radical openness to loving new people, new places, new problems,
new questions, new answers, new possibilities. Thus, to develop
wisdom requires additional “work” on making sense of our
internal and external contexts—relationships, history, vision,
positionality. Wise leaders understand that simply knowing is not
enough. Principals who act with courage could exhibit some of these
habits as they engage in leadership as love: • Demand better, demand justice • Laugh and cry alongside others • Express outrage rooted in care for others • Pursue dreams with faculty, staff, and parents • Listen to anger and disappointment and respond in non-
defensive, bridge-building ways • Cultivate a strong sense of collective commitment among
adults, among students, and within their communities g
If we continue to define what leadership that reflects wisdom
could look like, principals would exercise some of the following
habits of love: Courage, therefore, is what makes us become more than what we
are and what love convinces us we can be. As leaders it allows
us to leave the safe(r) confines of our individual existence and
become part of a global community. • Cultivate self-knowledge and awareness • Be attuned to care of self and others • Learn from experiences of others to create inclusive,
responsive, and sustainable spaces Courage Harmony is the tree bending in the breeze, yet snapping in
the midst of a storm. Harmony is bringing your entire self and
knowing that some will welcome while others will withdraw. It also means knowing how you bring your entire self in ways
that may embrace or harm. It is an active state of being that
requires strength, vulnerability, comfort with ambiguity, inquiry,
and openness. Studies that offer lenses for effective leadership often do not
address how frightening leadership is, especially in a role
that traditionally demands strength and displays of control. Allowing oneself to be vulnerable, to show fear or love, demands
tremendous courage in the face of uncertainty and in a context
of fetishizing quantifiable outcomes. Leaders might wonder how
they will support teachers in challenging the status quo if
they cannot control the outcomes of trying something different
from district norms. In response to community trauma, school
shootings, and natural disasters, leaders may wonder how they Indigenous elders would say that to walk this earth in
harmony with oneself, others, and nature is to be truly human. Harmony requires that we embrace all the facets of our humanity,
negative and positive, strengths and weaknesses, hopes and fear. January 2019 | Volume 3 | Article 117 Frontiers in Education | www.frontiersin.org 5 Byrne-Jiménez and Yoon Leading With Love What we know—or think we know—is an elusive thing. Knowing
the difference, including what remains to be known, is wisdom. It
is more than what we think of as smarts or intelligence, although
that can be an important piece. While knowledge derives from
interacting with acceptable texts and experts, where empirically
determined facts are the stock in trade; wisdom comes from
understanding your own journey, learning from the pain and
mistakes along the way, recognizing joy, and integrating those
experiences into your worldview. In this sense, wisdom comes
from developing a self-awareness and a self-love that is not
narcissistic but compassionate toward the self and others (Nhat
Hanh, 2015). Equally important is using that wisdom to ask
others if you can accompany them on their journey, if only for
a while. This offering is an externalization of love because it is
“knowing,” deep down in the recesses of our hearts, that our
journeys are connected—across time, space, generations—and
that the need to get there faster, ahead of others, is an empty
quest. Frontiers in Education | www.frontiersin.org Imagination Martin Luther King, Jr., in the face of the violence he saw
and experienced, imagined a different kind of world centered
in/around love and one in which everyone’s dreams were
precious. He believed in the possibility of this world so much that
he fought for justice through non-violent resistance despite—or
perhaps because of—the anger he must have felt every day of
his life. James Baldwin reminded us over and over that to be a
person of color in this country meant to be in a “constant state
of rage.” To lead for justice requires rage and hope that can only
be lifted up by the ability to imagine something new, better, and
more beautiful. In our own exploration we rely on the work of other scholars
and their re/framing of educational leadership. For example,
early research by Scheurich (1998) and Murtadha-Watts (1999)
began to introduce the role of love and spirituality within leader
practices. Dantley’s work on critical spirituality [Dantley (2003,
2005, 2010) i.e., 2003, 2005, 2010] and Alston’s on servant
(Alston, 2005) helped paved the way by making space in the
leadership discourse for our spirit and humanity. Our habits are
also linked to the work of other social justice scholars who helped
the field focus on creating equitable schools for minoritized and
under-served children. The critical analysis of existing school
structures (e.g., curriculum, policies) as barriers to students’ well-
being and life chances raised the alarm for scholars and school
leaders (Larson and Murtadha, 2002). Shields (2004) argued for
the need for transformative leaders whose work was grounded in
ethics and care. Such leaders overcome “pathologies of silence”
(p. 118), including the erasure of difference, through moral
dialogues that recognize the uniqueness of all individuals. More
recently, Theoharis (2007) defined school leadership as rooted
in justice, care, and empathy. A final area of synergy exists
with culturally responsive leadership, which highlights the need
for leaders to create inclusive schools for culturally diverse and
non-majoritarian students and families (Johnson, 2014; Khalifa,
2018). The “responsive” nature of leadership honors community
strengths, histories, differences, and needs. The scholars noted
here, connect the core of their vision of leadership with the
practices and purposes of schooling and education. In this sense, imagination is powerful and creative: powerful
enough to resist cynicism and creative enough to be truly free. Freedom is made of dreams, and dreams help us transgress what
already exists. Imagination Imagination is transcendent, while being equal
parts rebellious and visionary. When we imagine, as leaders,
we draw on our other habits of the heart. It takes courage and
wisdom to figure out how to dream, how to build. For school leaders, imagination helps them to see past the
limits and deficits of current ways of thinking about and
interacting with children, youth, families, and communities. This
then allows leaders to envision new ways of expressing their
dreams, of working with teachers and staff, and of making
schools centers of harmonious, courageous, and wise learning. This visioning builds on the individual and collective strengths
of the community. Imagination also makes room for empathy and understanding
of perspectives different from leaders’ own reality. As Maya
Angelou said, “If one is lucky, a solitary fantasy can totally
transform a million realities.” To walk in another’s shoes, to
imagine a different way of seeing the world, helps leaders to build
meaningful connections and tap into our shared humanity. It
allows leaders to embrace a sense of wonder and curiosity about
people, their thinking and learning, such that their behaviors
communicate value and respect for whole and complex persons. By cultivating a shared habit of imagining, leaders can help
people find where they belong. Perhaps most important is the
role of leaders in crafting and listening to the histories and
stories of individual students, teachers, staff, parents, and their
communities. These stories are sources of wisdom and identity, of
shoring up courage with reminders of past success or resilience. Too often our field debates the “rightness” of one perspective
over another, with little room for theory building and evolution. In an effort to resist this trend, we offer our habits of the heart
as concrete ways of understanding and demonstrating leadership
that centers the humanity, knowledge bases, and traditions of
individual leaders. Naming these habits allows us also to enter
leadership spaces with intentionality and our humanity intact. The final exploration of leadership as love would see principals
who engender habits of imagination because they: The final exploration of leadership as love would see principals
who engender habits of imagination because they: • Listen to and amplify the stories from the heart Wisdom • Listen with care and compassion • Understand that dreaming and acting are equally valuable I grew up hearing my mother say, “Sólo sé que no sé nada” (the
only thing I know is that I don’t know anything). There were
two important lessons embedded in that phrase: (1) knowledge is
constantly changing and (2) humility. It is only now that I recognize
these as wisdom—a wisdom handed down through generations of
my family and grounded in love. • Discern the relationships among internal and external politics,
resources, and hopes Wisdom is affirming in that it creates a different set of
relationships within ourselves, with others, with the past and
future, and with the environment. By embracing the limits of January 2019 | Volume 3 | Article 117 Frontiers in Education | www.frontiersin.org 6 Leading With Love Byrne-Jiménez and Yoon INTEGRATING THE HABITS OF LOVE individual knowledge leaders can approach their work with
humility and renewed purpose. In the previous section, we explored harmony, courage, wisdom,
and imagination as four habits of the heart that both define
leadership as love and provide a set of practices that exemplify
that leadership. We engaged in this “thought experiment” in
order to push our own theorizing of how love can be enacted
as a form of leadership. In our hearts and in our minds we
know that this is the beginning of an idea. In the same spirit
in which we developed this leadership framework—from a place
of openness and interconnection—we embrace the four habits
and their overlapping, mutually reinforcing energies. Rather than
developing another theory, we want our thinking to “live” in
harmony with existing leadership paradigms. Frontiers in Education | www.frontiersin.org CLOSING THOUGHTS ON LEADERSHIP AS
LOVE • Build and share dreams • Transgress the status quo • Play out different perspectives • Make intentional space for creativity and unstructured work • Make intentional space for creativity and unstructured work It is fitting that we ended our discussion of these leadership habits
of love with imagination, since we have asked you to imagine
a different way of understanding and enacting leadership. You
may also have noticed that we purposefully avoided some of Leading with imagination, then, is about being hopeful and
intentional about making collective dreaming a possibility for
children and schools. January 2019 | Volume 3 | Article 117 Frontiers in Education | www.frontiersin.org 7 Byrne-Jiménez and Yoon Leading With Love supervision and evaluation in K-12 settings were not created with
the habits of heart in mind. Nor were the promotion and tenure
or accreditation processes in higher education created with heart. These systems make us believe that education is a zero-sum game
and pit us against each other. In fact, they attack our humanity. In
resistance, we would need to rethink how we prepare and support
leaders in pre- and in-service learning contexts. We would also
need to develop new ways of engaging with policy makers and
politics that has both love and justice at the core. Lastly, we would
need to embrace indigenous and other emerging methodologies
that privilege harmony, wisdom, and multiple dimensions of
reality as ways to develop human-centered questions, theories,
and relationships. the “standard” language used to describe leadership. In contrast,
we tried using language steeped in the heart, in our imaginings
of a different way of leading. It is our imaginations, in fact,
that allowed us to engage in this thought experiment. You may
have also found yourself feeling awkward or uncomfortable or
asking about the evidence for this framework. We ask that you
ask yourself why you are asking that! What skepticisms make
a love-centered leadership seem impossible (or not) as a source
of knowledge and practice? We must ask ourselves what in our
training or discipline or assumptions makes a discussion of love
and leadership so difficult, irrelevant, or unrealistic. Ultimately, our invitation has been to engage in a thought
experiment as a way to reflect on our pasts as well as our
futures. AUTHOR CONTRIBUTIONS School leaders, however, cannot do it alone. Even as we have
focused our thinking on the school principal, we know that there
are larger systems and more powerful actors that also need to be
a part of this new framing of leadership. Our current systems of MB-J and IY were equal contributors to the conceptualization
and writing of this essay. CLOSING THOUGHTS ON LEADERSHIP AS
LOVE Don’t even over-
sentimentalize it. There’s power, power in love. - Reverend Michael
Curry There’s power in love. Don’t underestimate it. Don’t even over-
sentimentalize it. There’s power, power in love. - Reverend Michael
Curry Curry CLOSING THOUGHTS ON LEADERSHIP AS
LOVE In our individual and collective histories we must look
for the stories of harmony, wisdom, and courage that, if we have
been fortunate, have shaped our understandings of leadership. And in those stories, where were we allowed or encouraged to
imagine a different way of being for ourselves and others? In
what contexts did those stories emerge? How do we reshape
and amplify those stories in our lives, personal and professional? What does embracing love as an active practice afford leaders in
complex and sometimes painful situations? As you can see, there is more work to be done to make this
framework a reality. We are researchers, after all, and we cannot
stay at the level of conceptualization when there are children
and school communities in pain. We want to recognize and
work with leaders who practice the habits of love to deepen our
understanding of how this leadership survives in and perhaps
transforms educational systems. More
importantly,
we
want
to
continue
our
inward
exploration, as women of color who prepare the next generation
of educational leaders and scholars, and how our histories
strengthen and shape our own leadership. It is in the echoes
of our pasts that we find our way forward. This journey to
re/claim our stories and voices is as much an essential part of
our scholarship as it is of our leadership. Because of this, we are
confident that leadership is an act of deep, boundless, ancient,
powerful love. We invite you to join us in this work! Every day leaders are called upon to make decisions
that impact the lives of children, families, teachers, and
staff. Increasingly, school leaders are standing in the way of
Immigration and Customs Enforcement officers or shielding
their students in the aftermath of another school/community
shooting or nurturing the fragile joy and promise of young people
in the face of curriculum and assessments that alienate them. Every day, school leaders must choose between what is right
and what is good, understanding that what may be right by
organizational standards may not be good for students and their
families. Without a clear and firm grounding in the habits of love,
leaders may very well lose their humanity in these situations. There’s power in love. Don’t underestimate it. Don’t even over-
sentimentalize it. There’s power, power in love. - Reverend Michael
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Compassion, and Character at School. Alexandria, VA: ASCD. Yaron, L. (2015). Mindfulness in the Classroom: A How-to Guide. Education
Week. Available online at: https://www.edweek.org/tm/articles/2015/11/10/
mindfulness-in-the-classroom-a-how-to-guide.html (Accessed November 10,
2015). Khalifa, M. (2018). Culturally Responsive School Leadership. Cambridge, MA:
Harvard Education Press. King, M. L. Jr. (1967). Beyond Vietnam: A Time to Break Silence. Speech delivered
at Riverside Church, New York, NY. Available online at: https://kinginstitute. stanford.edu/king-papers/documents/beyond-vietnam
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1967). Yoon, I. H. (2017). Considering the Future of U.S. Public Schooling: President-
Elect Trump, ESSA, and Educational Leadership. Division A Newsletter of
the American Educational Research Association. Available online at: http://
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J. C. (2000). A
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epistemology:
what
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hold
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my life, faith, and profession. Int. J. Qual. Stud. Edu. 13, 683–692. doi: 10.1080/09518390050211574 Conflict of Interest Statement: The authors declare that the research was
conducted in the absence of any commercial or financial relationships that could
be construed as a potential conflict of interest. Meyerson, D. E., and Scully, M. A. (1995). Crossroads tempered radicalism and the
politics of ambivalence and change. Organ. Sci. 6, 585–600. Copyright © 2019 Byrne-Jiménez and Yoon. This is an open-access article distributed
under the terms of the Creative Commons Attribution License (CC BY). The use,
distribution or reproduction in other forums is permitted, provided the original
author(s) and the copyright owner(s) are credited and that the original publication
in this journal is cited, in accordance with accepted academic practice. No use,
distribution or reproduction is permitted which does not comply with these terms. Murtadha-Watts, K. (1999). REFERENCES Dantley, M. E. (2003). Critical spirituality: enhancing transformative leadership
through critical theory and African American prophetic spirituality. Int. J. Leadership Edu. 6, 3–17. doi: 10.1080/1360312022000069987 Alisic, E. (2012). Teachers’ perspectives on providing support to children after
trauma: a qualitative study. Sch. Psychol. Q. 27, 51–59. doi: 10.1037/a0028590 Dantley, M. E. (2005). African American spirituality and Cornel West’s notions
of prophetic pragmatism: restructuring educational leadership in American
urban schools. Edu. Administration Q. 41, 651–674. doi: 10.1177/0013161x042
74274 Alston, J. A. (2005). Tempered radicals and servant leaders: black females
persevering in the superintendency. Edu. Administration Q. 41, 675–688. doi: 10.1177/0013161X04274275 Dantley, M. E. (2010). Successful leadership in urban schools: principals and
critical spirituality, a new approach to reform. J. Negro Edu. 79, 214–219. Available online at: http://www.jstor.org/stable/20798344 Byrne-Jiménez, M. C. (2017). 2016 Presidential Address: leading in dangerous
times: leadership as an act of love. UCEA Rev. Winter 2017, 1–5. Capper, C. (2000). Life lessons and a loving epistemology: a response to
Julie Liable’s loving epistemology. Int. J. Qual. Stud. Edu. 13, 693–698. doi: 10.1080/09518390050211583 Dantley, M. E., and Tillman, L. C. (2010). “Social justice and moral transformative
leadership,” in Leadership for Social Justice: Making Revolutions in Education, ed
C. Marshall and M. Oliva (Boston, MA: Allyn & Bacon Publishers), 19–34. Coldewey, D. (2018). Facebook Invented a New Time Unit Called the “Flick” and
It’s Truly Amazing. TechCrunch. Available online at: https://techcrunch.com/
2018/01/22/facebook-invented-a-new-time-unit-called-the-flick-and-its-
truly-amazing/ (Accessed January 22, 2018). Darder, A. (1998). Teaching as an Act of Love: Reflections on Paulo Freire and
His Contributions to Our Lives and Work. San Bernadino, CA: California
Association for Bilingual Education. Conrad, D., and Kellar-Guenther, Y. (2006). Compassion fatigue, burnout, and
compassion satisfaction among Colorado child protection workers. Child Abuse
Neglect. 30, 1071–1080. doi: 10.1016/j.chiabu.2006.03.009 Duncan-Andrade, J. M. R., and Morrell, E. (2008). The Art of Critical Pedagogy:
Possibilities for Moving from Theory to Practice in Urban Schools. New York,
NY: Peter Lang. doi: 10.3726/b12771 January 2019 | Volume 3 | Article 117 Frontiers in Education | www.frontiersin.org 8 Byrne-Jiménez and Yoon Leading With Love Nee-Benham, A., Maenette, K. P., and Cooper, J. E. (1998). Let My Spirit Soar! Narratives of Diverse Women in School Leadership. Thousand Oaks, CA: Corwin
Press. Durlak, J. A., Weissberg, R. P., Dymnicki, A. B., Taylor, R. D., and Schellinger, K. B. (2011). The impact of enhancing students’ social and emotional learning: a
meta-analysis of school-based universal interventions. Child Dev. 82, 405–432. REFERENCES doi: 10.1111/j.1467-8624.2010.01564.x Nhat Hanh, T. (2015). How to Love. Berkeley, CA: Parallax Press. Elmore, R. F., and Burney, D. (1997). Investing in Teacher Learning: Staff
Development and Instructional Improvement: Community School District 2,
New York City. New York, NY: National Commission on Teaching and
America’s Future and the Consortium for Policy Research in Education. Elmore, R. F., and Burney, D. (1997). Investing in Teacher Learning: Staff
Development and Instructional Improvement: Community School District 2, Overvold, G. E. (1987). The imperative of organizational harmony: a critique
of contemporary human relations theory. J. Business Ethics 6, 559–565. doi: 10.1007/BF00383747 New York City. New York, NY: National Commission on Teaching and
America’s Future and the Consortium for Policy Research in Education. Palmer, P. J. (1983). To Know as We are Known: Education as a Spiritual Journey. New York, NY: HarperCollins Publishers. Freire, P. (1970). Pedagogy of the Oppressed. New York, NY: Bloomsbury
Publishing. Poplin, M. (1992). The leader’s new role: looking to the growth of teachers. Edu. Leadership 49, 10–11. Freire, P. (1998). Teachers as Cultural Workers: Letters to Those Who Dare Teach. Boulder, CO: Westview. Scheurich, J. J. (1998). Highly successful and loving, public elementary schools
populated mainly by low-SES children of color: core beliefs and cultural
characteristics. Urban Edu. 33, 451–491. doi: 10.1177/0042085998033004001 Hooks, B. (1994). Teaching to Transgress: Education as the Practice of Freedom. New
York, NY: Routledge. g
Hooks, B. (2001). All About Love: New Visions. New York, NY: Harper Perennial. Hooks, B. (2001). All About Love: New Visions. New York, NY: Harper Perennial. Hooks B (2009) Belonging: A Culture of Place New York NY: Routledge Shaia, W. E., and Crowder, S. C. (2017). “Schools as retraumatizing environments,”
in Linking Health and Education for African American Students’ Success, ed N. M Finigan Carr (New York NY: Routledge) 69 82 Hooks, B. (2009). Belonging: A Culture of Place. New York, NY: Routledge. Hooks, B. (2009). Belonging: A Culture of Place. New York, NY: Ro Johnson,
L. (2014). Culturally
responsive
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empowerment. Multicult. Educ. Rev. 6, 145–170. Shields, C. M. (2004). Dialogic leadership for social justice: overcoming
pathologies
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Q. 40,
109–132. doi: 10.1177/0013161X03258963 p
Kay, K., and Greenhill, V. (2013). The Leader’s Guide to 21st Century Education: 7
St p f
S h
l
d Di t i t B
t
MA P
Ed
ti p
Kay, K., and Greenhill, V. (2013). Frontiers in Education | www.frontiersin.org January 2019 | Volume 3 | Article 117 REFERENCES “Spirited sisters: spirituality and the activism of
African American women in educational leadership,” in School Leadership:
Expanding the Horizons of the Mind and Spirit: 7th Yearbook of the
National Council of Professors of Educational Administration, ed L. T. Fenwick
(Lancaster, PA: Technomic Publishing Company), 155–167. (Lancaster, PA: Technomic Publishing Company), 155–167. January 2019 | Volume 3 | Article 117 Frontiers in Education | www.frontiersin.org 9
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FDA Approved Drugs Efavirenz, Tipranavir, and Dasabuvir Inhibit Replication of Multiple Flaviviruses In Vitro
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None
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English
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Abstract
FDA Approved Drugs Efavirenz, Tipranavir, and
Dasabuvir Inhibit Replication of Multiple
Flaviviruses In Vitro †
Michal Stefanik 1,2,*, Fortunatus C Ezebuo 3, Jan Haviernik 1,4, Ikemefuna C. Uzochukwu 3,
Martina Fojtikova 1, Jiri Salat 1, Ludek Eyer 1,5 and Daniel Ruzek 1,5
Department of Virology, Veterinary Research Institute, Hudcova 70, CZ-62100 Brno, Czech Republic;
haviernik@vri.cz (J.H.); fojtikova@vri.cz (M.F.); salat@vri.cz (J.S.); eyer@vri.cz (L.E.);
ruzekd@paru.cas.cz (D.R.)
2 Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno,
Czech Republic
3 Department of Pharmaceutical and Medicinal Chemistry, Faculty of Pharmaceutical Sciences,
Nnamdi Azikiwe University, PMB 5025 Awka 420281, Nigeria; fc.ezebuo@unizik.edu.ng (F.C.E.);
ic.uzochukwu@unizik.edu.ng (I.C.U.)
4 Faculty of Science, Masaryk University, Kamenice 5, 625 00 Brno, Czech Republic
5 Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, Branisovska 31,
CZ-37005 Ceske Budejovice, Czech Republic
* Correspondence: michal.stefanik2@gmail.com
† Presented at Viruses 2020—Novel Concepts in Virology, Barcelona, Spain, 5–7 February 2020.
1
Published: 2 June 2020
Abstract: Arthropod-borne flaviviruses such as tick-borne encephalitis virus (TBEV), West Nile
virus (WNV), Zika virus (ZIKV), Dengue virus (DENV), and yellow fever virus (YFV) cause several
serious life-threatening syndromes (encephalitis, miscarriages, paralysis, etc.). No effective antiviral
therapy against these viruses has been approved yet. We selected, via in silico modeling, 12 U.S.
Food and Drug Administration (FDA)-approved antiviral drugs (paritaprevir, dolutegravir,
raltegravir, efavirenz, elvitegravir, tipranavir, saquinavir, dasabuvir, delavirdine, maraviroc,
trifluridine, and tauroursodeoxycholic acid) for their interaction with ZIKV proteins (NS3 helicase
and protease, non-structural protein 5 (NS5) RNA-dependent RNA polymerase, and
methyltransferase). Only three of them were active against ZIKV, namely, dasabuvir (ABT-333),
efavirenz, and tipranavir. These compounds inhibit virus replication of ZIKV (MR-766 and
Paraiba_01) in Vero cells; therefore, we tested these compounds against other medically important
flaviviruses WNV (13-104 and Eg101) and TBEV (Hypr). Dasabuvir was originally developed as an
antiviral drug against hepatitis C virus (HCV); tipranavir and efavirenz are used for treating human
immunodeficiency virus (HIV) infection. The antiviral effects of efavirenz, tipranavir, and dasabuvir
were tested for ZIKV in HUH-7, astrocytes (HBCA), and UKF-NB-4 cells, where we also identified
a significant inhibition effect of these compounds. For Vero cells, efavirenz inhibited all investigated
viruses with EC50 ranging from 9.70 to 29.26 µM; the tipranavir inhibition effect was from 16.19
(WNV 13-104) to 27.47 µM (TBEV), while the strongest and most robust antiviral effect was
demonstrated in the case of dasabuvir (EC50 values ranging from 9.09 (TBEV) to 10.85 µM (WNV
13-104)). These results warrant further research of these drugs, either individually or in
combination, as possible pan-flavivirus inhibitors.
Proceedings 2020, 50, 6; doi:10.3390/proceedings2020050006
www.mdpi.com/journal/proceedings
Proceedings 2020, 50, 6
2 of 2
Keywords: flaviviruses; dasabuvir; tipranavir; efavirenz; antiviral drug
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).
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Confiance élevée et modérément élevée dans le gouvernement national, les collectivités locales, la fonction publique, la justice et le parlement, 2021
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12 https://stat.link/khvcuz
Panorama des administrations publiques 2023 © OCDE 2023
161
10. RECETTES PUBLIQUES ET COÛTS DE PRODUCTION
Coûts de production et externalisation
Les coûts de production des administrations publiques sont
les dépenses publiques consacrées aux biens et services que
ces administrations utilisent. Ils recouvrent la rémunération
des agents publics (salaires) et les achats de biens et de
services (fournitures pour les écoles et les hôpitaux par
exemple). Ils ne comprennent pas les dépenses publiques qui
n’impliquent pas l’achat d’un bien ou d’un service (comme
les dépenses liées à la protection sociale, les allocations de
chômage et d’autres transferts). L’externalisation est la partie
des coûts de production des administrations publiques qui est
utilisée pour acheter des biens et services à des entités non
gouvernementales (achats des administrations publiques à des
entreprises privées et à d’autres organismes).
En 2021, les coûts de production ont représenté en moyenne
à 21.6 % du PIB dans les pays de l’OCDE (graphique 10.12). Les
plus élevés ont été enregistrés en Finlande (31.1 %), en Islande
(30.3 %) et en Suède (29.5 %) - tous des pays scandinaves -,
ce qui s’explique par la prestation généralisée de services à
financement public et leurs coûts relativement élevés. Les pays
qui ont consacré la plus faible part de leur PIB à ces dépenses
sont le Mexique (11.8 %), la Colombie (16.8 %) et le Chili
(16.8 %). Les coûts de production des administrations publiques
ont diminué dans 25 des 27 pays pour lesquels des données
étaient disponibles en 2022, de 1.1 % du PIB en moyenne. Cette
baisse tient sans doute à ce que certains des services publics
fournis en réponse à la pandémie de COVID-19 n’étaient plus
nécessaires, ou à la croissance du PIB qui a accompagné le
redressement des pays, ou aux deux à la fois.
La composition des coûts de production varie légèrement selon
les pays de l’OCDE (graphique 10.13). Les dépenses consacrées
à la rémunération des agents publics en ont représenté en
moyenne 43.2 %, ce pourcentage se situant entre 40 % et 55 %
dans la plupart des pays de l’OCDE (28 sur 37). Deux pays
ont alloué à ce poste une part nettement supérieure de leurs
dépenses : le Mexique (72.7 %) et le Costa Rica (70.9 %). Les
pays qui y ont consacré la part la plus faible sont le Japon
(23.5 %) et les Pays-Bas (29.4 %). Les dépenses salariales ne sont
pas nécessairement liées à la structure des administrations
publics. Par exemple, l’Irlande (47.1 %) et le Canada (48.8 %)
y ont affecté des montants quasiment identiques, alors que
l’Irlande a une administration publique unitaire et centralisée
et le Canada un système fédéral. Les dépenses consacrées à
l’achat de biens et de services ont constitué en moyenne 44.1 %
des coûts de production, ce pourcentage étant compris entre
30 % et 45% dans la majorité des pays de l’OCDE (24 sur 37).
Les administrations publiques ont consacré en moyenne 9.5 %
du PIB aux dépenses d’externalisation en 2021 (graphique 10.14).
Sur ce total, 6.2 % du PIB ont été alloués au paiement d’acteurs
non gouvernementaux pour la fourniture de biens et de services
utilisés directement par les administrations publiques. 3.3 %
du PIB à des biens et des services fournis à la population
par des prestataires non gouvernementaux, mais financés
par les pouvoirs publics, dans les domaines de la santé, du
logement, du transport et de l’éducation par exemple. Dans
22 des 27 pays pour lesquels on dispose de données, les coûts
d’externalisation ont notablement diminué en 2022, de 0.3 %
du PIB en moyenne. Les différentes structures de dépenses
peuvent refléter des décisions nationales différentes quant à la
fourniture directe de biens et de services par le gouvernement
ou à leur externalisation. Par exemple, tandis que les deux
gouvernements sont de taille équivalente en part du PIB, les
Pays-Bas dépensent beaucoup plus pour financer les biens et
services fournis au public par des sous-traitants (10.5 % du
PIB) que le Danemark (1.2 % du PIB) (figure 10.14). De même, le
Danemark consacre une part beaucoup plus importante de ses
162
coûts de production liés à la rémunération des agents publics
(53.7 %) que les Pays-Bas (29.4 %) (graphique 10.13).
Méthodologie et définitions
Le concept et la méthode de calcul des coûts de
production reposent sur la classification des dépenses
des administrations publiques propre au Système de
comptabilité nationale (SCN). Le cadre du SCN 2008 est
désormais en vigueur dans tous les pays de l’OCDE
(voir l’annexe C pour un complément d’information).
Les coûts de production des administrations publiques
comprennent : Les coûts liés à la rémunération
des agents publics, y compris les rémunérations en
espèces ou en nature, plus la totalité des cotisations
obligatoires (et imputées) de l’employeur au régime
d’assurance sociale et les versements facultatifs pour
le compte des agents publics. Les biens et les services
utilisés par les administrations publiques, qui sont la
première composante de l’externalisation. Selon le SCN,
il s’agit de la consommation intermédiaire (achat des
produits intermédiaires nécessaires à la production des
administrations). Les biens et les services financés par
les administrations publiques, deuxième composante
des dépenses d’externalisation. Selon le SCN, cette
composante inclut les transferts sociaux en nature
effectués par l’intermédiaire de producteurs marchands et
payés par les administrations publiques. Les autres coûts
de production, qui englobent les composantes restantes
de la consommation de capital fixe (amortissement du
capital) et les autres impôts sur la production, moins les
autres subventions à la production. Les données couvrent
l’emploi public et la consommation intermédiaire
affectés à des activités de production destinées à l’usage
propre des administrations publiques. Les coûts de
production présentés ici ne sont pas égaux à la valeur
de la production dans le SCN.
Pour en savoir plus
OCDE (2022), OECD Economic Surveys: Finland 2022, Éditions
OCDE, Paris, https://doi.org/10.1787/516252a7-en.
OCDE (2022), OECD Economic Surveys: Mexico 2022, Éditions
OCDE, Paris, https://doi.org/10.1787/2e1de26c-en.
Notes relatives aux graphiques
Les données concernant Türkiye ne sont pas comprises dans la moyenne
OCDE.
10.12. Les données relatives au Chili ne sont pas comprises dans la
moyenne OCDE. Celles concernant le Brésil, l’Indonésie et Türkiye
portent sur 2020 et non 2021.
10.13. Les données concernant le Chili ne sont pas disponibles. Celles
concernant l’Indonésie et Türkiye portent sur 2020 et non 2021.
10.14. Les données pour le Chili ne sont pas disponibles. L’Afrique du
Sud, les États-Unis, l’Indonésie et le Mexique ne présentent pas
séparément les biens et les services financés par les administrations
publiques dans leurs comptes nationaux. Les données pour le Brésil,
l’Indonésie et Türkiye portent sur 2020 et non 2021.
G.5.5 (Composition des dépenses d’externalisation des administrations
publiques, 2021) est consultable en ligne dans l’annexe G.
Panorama des administrations publiques 2023 © OCDE 2023
10. RECETTES PUBLIQUES ET COÛTS DE PRODUCTION
Coûts de production et externalisation
10.12. Coûts de production en pourcentage du PIB, 2021 et 2022
%
35
Rémunération des agents publics
Coûts des biens et services utilisés et financés par les administrations publiques
Autres coûts de production
Rémunération des agents publics, 2022
Coûts des biens et services utilisés et financés par les administrations publiques, 2022
Autres coûts de production, 2022
30
25
20
15
10
5
BR
A
HR
V
ZA
F
BG
R
RO
U
ID
N
FI
N
IS
SW L
E
NL
D
FR
A
DN
K
BE
L
NO
R
CA
N
AU
T
O C DE
DE U
-U
E
AU
S
GB
R
GR
C
HU
N
CZ
E
SV
N
NZ
L
LV
A
IS
R
SV
K
ES
P
JP
N
ES
T
IT
A
PR
OC T
DE
PO
L
LU
X
KO
R
LT
U
US
A
TU
R
CR
CH I
E
CO
L
IR
L
M
EX
CH
L
0
Source : Statistiques de l’OCDE sur les comptes nationaux (base de données). Pour l’Australie, les données sont tirées à la fois des statistiques sur les
comptes nationaux et des statistiques sur les finances publiques communiquées par le Bureau australien des statistiques.
12 https://stat.link/7zmq6t
10.13. Composition des coûts de production, 2021
Rémunération des agents publics
Coûts des biens et services utilisés et financés par les administrations publiques
Autres coûts de production
%
100
80
60
40
20
ZA
F
RO
U
ID
N
BG
R
HR
V
M
EX
CR
I
LT
U
DN
K
PR
T
SV
N
IS
L
ES
P
NO
R
LU
X
PO
L
GR
C
US
A
CA
N
ES
T
SV
K
LV
A
TU
R
IR
L
BE
L
CZ
E
CH
E
FR
A
CO
L
IT
A
HU
N
AU
T
SW
E
GB
R
FI
N
IS
R
NZ
L
KO
R
AU
S
DE
U
NL
D
JP
N
OC OCD
DE E
-U
E
0
Source : Statistiques de l’OCDE sur les comptes nationaux (base de données). Pour l’Australie, les données sont tirées à la fois des statistiques sur les
comptes nationaux et des statistiques sur les finances publiques communiquées par le Bureau australien des statistiques.
12 https://stat.link/3rkz7f
10.14. Dépenses d’externalisation des administrations publiques en pourcentage du PIB, 2021 et 2022
%
20
Biens et services utilisés par les administrations publiques, 2021
Biens et services financés par les administrations publiques, 2021
Biens et services utilisés par les administrations publiques, 2022
Biens et services financés par les administrations publiques, 2022
16
12
8
4
BR
A
HR
V
BG
R
ZA
RO F
U
ID
N
NL
DE
D
U
FI
N
AU
S
JP
N
GB
R
NZ
L
BE
L
IS
L
AU
T
OC FR
DE A
-U
SW E
E
IS
R
DN
K
HU
N
CA
N
KO
OC R
DE
NO
R
CZ
E
SV
K
LV
A
SV
N
ES
P
IT
A
GR
C
ES
T
CO
L
PO
L
PR
T
LU
X
TU
R
US
A
CH
E
LT
U
IR
L
CR
M I
EX
0
Source : Statistiques de l’OCDE sur les comptes nationaux (base de données). Pour l’Australie, les données sont tirées à la fois des statistiques sur les
comptes nationaux et des statistiques sur les finances publiques communiquées par le Bureau australien des statistiques.
12 https://stat.link/e3y9io
Panorama des administrations publiques 2023 © OCDE 2023
163
11. DÉPENSES PUBLIQUES
Dépenses des administrations publiques
Ventilation des dépenses des administrations publiques
par fonction (CFAP/COFOG)
Ventilation des dépenses publiques liées aux fonctions
de protection sociale et de santé (CFAP/COFOG)
Rapport coût-efficacité
Ventilation des dépenses des administrations publiques
par opération économique
Ventilation des dépenses des administrations publiques selon
les niveaux d’administration
Dépenses d’investissement des administrations publiques
Solde budgétaire des administrations publiques
Solde structurel des administrations publiques
Réduction des inégalités et de la pauvreté
Panorama des administrations publiques 2023 © OCDE 2023
165
11. DÉPENSES PUBLIQUES
Dépenses des administrations publiques
Les administrations publiques assurent la fourniture de tout
un éventail de biens et de services à la population. Certains de
ces biens et services relèvent de leur compétence exclusive ;
tel est notamment le cas du système judiciaire. D’autres,
comme les soins de santé, peuvent être fournis à la fois par des
acteurs publics et par des acteurs privés. En plus d’assurer des
services, les administrations publiques s’emploient à assurer
une redistribution des revenus à l’échelle de la société, au
moyen de prestations et d’aides sociales. La place qu’occupent
les acteurs publics dans la fourniture de biens et de services
varie nettement selon les pays, en fonction de leurs choix
stratégiques, de leurs priorités du moment et de leurs systèmes
et traditions politiques. Sur l’ensemble de la zone OCDE, les
dépenses publiques sont principalement consacrées à la
fourniture de services publics et à la réalisation de transferts
de revenus. Elles tendent à être plus stables que les recettes
publiques, qui dépendent davantage des cycles économiques.
Grâce aux dépenses publiques, les administrations offrent à la
population un filet de sécurité fiable, en lui ouvrant certains
droits et en amortissant les fluctuations économiques.
En 2021, en moyenne de la zone OCDE, les dépenses des
administrations publiques ont représenté 46.3 % du PIB. C’est
en France (59.1 %), en Grèce (57.7 %) et en Italie (57.3 %) que
les pourcentages étaient les plus élevés. Comme le montre
le graphique 11.1, le poids des dépenses des administrations
publiques dans le PTB a progressé de 5.4 points de pourcentage
(p.p.) entre 2019 (où il s’établissait à 40.9 %) et 2021. Cette
augmentation s’explique en grande partie par la pandémie de
COVID-19, qui a bouleversé la vie économique. Les autorités
ont réagi en lançant de vastes plans de relance budgétaire, et
elles ont notamment augmenté leurs dépenses en matière de
santé, de protection sociale et de soutien aux entreprises et
aux individus touchés par la pandémie ; dans le même temps,
le PIB reculait.
En 2020, sous l’effet de la pandémie, le poids des dépenses
publiques a atteint un sommet dans la zone OCDE (à 48.4 %
du PIB en moyenne) et dans les plus grandes économies de
la zone, avant de retomber en 2021 et 2022. Ce poids reste
toutefois beaucoup plus important qu’avant le COVID-19
(graphique 11.2). Dans 35 pays de l’OCDE sur 38, le poids
des dépenses publiques dans le PIB a progressé entre 2019
et 2021. C’est en Grèce (+ 9.6 p.p.) et en Italie (+8.8 p.p.) que
l’augmentation a été la plus forte. Entre 2021 et 2022, le poids
des dépenses publiques dans le PIB a baissé dans 26 des
27 pays pour lesquels on dispose de données ; le Luxembourg
fait figure d’exception, avec une augmentation de 0.4 p.p.
(graphique 11.1).
En 2021, en moyenne de la zone OCDE, les dépenses des
administrations publiques se sont élevées à 23 432 USD à PPA
par habitant ; les montants s’échelonnaient entre 5 637 USD à
PPA au Mexique et 56 357 USD à PPA au Luxembourg. Entre 2019
et 2021, les dépenses par habitant ont augmenté, en moyenne,
de 3 695 USD à PPA. C’est aux États-Unis (+6 663 USD à PPA) et
au Luxembourg (+4 925 USD à PPA) que la hausse a été la forte
(graphique 11.3).
En moyenne de la zone OCDE, les dépenses publiques par
habitant ont augmenté de 12.5 % en termes réels en 2020.
166
En 2021, cette hausse s’est nettement ralentie, avec un taux
de croissance moyen de +0.64 % en termes réels. Dans les pays
pour lesquels on dispose de données, les dépenses par habitant
se sont mises à chuter en 2022 : 21 pays sur 27 ont enregistré
une baisse, qui a été particulièrement marquée en Norvège
(-17.0 %) (graphique G.6.1 en ligne).
Méthodologie et définitions
Les données relatives aux dépenses des administrations
publiques sont tirées de la base de données des
Statistiques de l’OCDE sur les comptes nationaux, qui sont
établies conformément au Système de comptabilité nationale
(SCN) ; ce dernier constitue un ensemble de concepts, de
définitions, de nomenclatures et de règles approuvés au
plan international en matière de comptabilité nationale.
Le cadre du SCN 2008 a été mis en œuvre par tous les
pays de l’OCDE (voir l’annexe C pour plus de précisions
sur les systèmes de déclaration et les sources). Selon
la terminologie du SCN, les administrations publiques
se composent de l’administration centrale, des
administrations d’États fédérés, des administrations
locales et des administrations de sécurité sociale. Les
dépenses englobent la consommation intermédiaire,
la rémunération des salariés, les subventions, les
revenus de la propriété (dont les paiements d’intérêts),
les prestations sociales, les autres dépenses courantes
(essentiellement les transferts courants) et les dépenses
en capital (transferts en capital et investissements). Le
produit intérieur brut (PIB) est la mesure habituelle de
la valeur des biens et services produits par un pays au
cours d’une période donnée. On a calculé les dépenses
publiques par habitant en convertissant les dépenses
publiques totales en USD aux parités de pouvoir d’achat
(PPA) OCDE/Eurostat pour le PIB et en divisant le résultat
obtenu par la population du pays. La PPA correspond
au nombre d’unités monétaires du pays B nécessaire à
l’acquisition d’une même quantité de biens et de services
dans le pays A.
Pour en savoir plus
OCDE (2023), Perspectives économiques de l’OCDE, Rapport
intermédiaire mars 2023, Une reprise fragile, Éditions OCDE,
Paris, https://doi.org/10.1787/2d7536fc-fr.
Notes relatives aux graphiques
Les données sur le Chili et Türkiye ne sont pas prises en compte dans
la moyenne de la zone OCDE.
11.1 et 11.3. Les données sur Türkiye, le Brésil et l’Indonésie portent
sur 2020, et non 2021.
G.6.1 (Taux annuel de progression en termes réels des dépenses
publiques par habitant, 2019-20, 2020-21 et 2021-22) est consultable
en ligne dans l’annexe G.
Panorama des administrations publiques 2023 © OCDE 2023
11. DÉPENSES PUBLIQUES
Dépenses des administrations publiques
11.1. Dépenses des administrations publiques en pourcentage du PIB, 2019, 2021 et 2022
2019
%
80
2021
2022
70
60
50
40
30
20
10
FR
A
GR
C
IT
A
AU
T
FI
N
OC B
D E EL
-U
E
DE
U
DN
K
ES
P
IS
L
SV
N
SW
E
GB
R
CO
L
HU
N
NO
R
PR
T
NL
D
CA
N
CZ
E
SV
OC K
DE
NZ
L
US
A
JP
N
LV
A
PO
L
CR
LU I
X
ES
T
IS
R
AU
S
KO
R
LT
U
CH
E
TU
R
CH
L
M
EX
IR
L
BR
A
HR
V
BG
R
RO
U
ID
N
0
Source : Statistiques de l’OCDE sur les comptes nationaux (base de données).
StatLink 2 https://stat.link/bq3drp
11.2. Dépenses des administrations publiques en pourcentage du PIB : zone OCDE et principales économies
de l’OCDE, 2007 à 2022
OCDE
%
55
OCDE-UE
JPN
GBR
USA
50
45
40
35
30
25
20
2007
2008
2009
2010
2011
2012
2013
2014
2015
2016
2017
2018
2019
2020
2021
2022
Source : Statistiques de l’OCDE sur les comptes nationaux (base de données).
StatLink 2 https://stat.link/4wpk8u
11.3. Dépenses des administrations publiques par habitant, 2019, 2021 et 2022
2019
2021
2022
HR
V
RO
U
BG
R
BR
A
ID
N
LU
X
NO
R
AU
T
DN
K
BE
L
US
A
FI
N
FR
A
DE
U
SW
E
NL
D
IS
L
CH
E
IT
A
O C IR
DE L
-U
E
CA
N
AU
S
GB
OC R
DE
SV
N
NZ
L
CZ
E
ES
P
JP
N
GR
C
ES
T
IS
R
KO
R
HU
N
PR
T
PO
L
LT
U
SV
K
LV
A
TU
R
CR
CH I
L
CO
M L
EX
USD à PPA
65 000
60 000
55 000
50 000
45 000
40 000
35 000
30 000
25 000
20 000
15 000
10 000
5 000
0
Source : Statistiques de l’OCDE sur les comptes nationaux (base de données).
StatLink 2 https://stat.link/v25qeu
Panorama des administrations publiques 2023 © OCDE 2023
167
11. DÉPENSES PUBLIQUES
Ventilation des dépenses des administrations publiques par fonction
(CFAP/COFOG)
Il appartient aux administrations publiques de financer ou
d’assurer directement un large éventail de services et d’activités
dans des domaines tels que la santé, l’éducation ou la justice ;
de garantir l’ordre public et la sécurité de la population civile ;
et de représenter leur pays sur la scène internationale. La
ventilation des dépenses publiques par fonction offre une
vue d’ensemble de l’utilisation des ressources publiques dans
des domaines clés. Elle éclaire aussi les priorités des pays
et leurs préférences quant aux modes d’exécution de leurs
missions (avec des solutions, soit intégralement publiques,
soit combinant interventions publiques et privées). Cette
ventilation peut évoluer en fonction de choix stratégiques,
mais aussi sous l’effet des tendances socioéconomiques à
l’œuvre (en matière de démographie, par exemple), des cycles
conjoncturels ou encore de certains chocs comme celui de la
pandémie de COVID-19.
En 2021, sur l’ensemble de la zone OCDE, c’est la protection
sociale qui représentait le plus grand poste de dépenses
des administrations publiques (tableau 11.4) ; les dépenses
correspondantes équivalaient, en moyenne, à 15.8 % du PIB.
Ce pourcentage s’échelonnait entre 8.7 % du PIB en Irlande
et 24.8 % du PIB en France. La protection sociale recouvre des
prestations en matière de vieillesse, de maladie, d’invalidité et
de chômage. Les pays de l’OCDE qui sont également membres
de l’Union européenne consacrent, en moyenne, 20.6 % de leur
PIB à la protection sociale, ce qui est supérieur à la moyenne
OCDE dans son ensemble.
La santé représente le deuxième poste de dépenses publiques.
Elle englobe les dépenses liées aux services hospitaliers, aux
services ambulatoires et aux produits, appareils et matériels
médicaux. Entre 2019 et 2021, les dépenses de santé sont
passées, en moyenne, de 8.0 % à 9.0 % du PIB dans la zone
OCDE, sous l’effet, notamment, de la pandémie de COVID-19.
En 2021, sur l’ensemble de la zone OCDE, ce sont les États-Unis
(10.3 %) et l’Autriche (10.1 %) qui ont affiché les pourcentages
les plus élevés dans cette catégorie. À l’inverse, la Suisse (2.8 %)
et le Luxembourg (5.4 %) ont affiché les pourcentages les plus
faibles (tableau 11.4). Pour la Suisse, le poids relativement peu
élevé des dépenses s’explique par la prédominance des acteurs
privés dans l’assurance maladie.
Les affaires économiques ont représenté le troisième poste
de dépenses pour la zone OCDE en 2021. Elles englobent les
dépenses réalisées par les administrations publiques dans les
domaines des échanges, de l’agriculture, de l’énergie et des
transports pour appuyer les activités productives. En moyenne,
les pays de l’OCDE ont consacré 5.7 % de leur PIB aux affaires
économiques ; les pourcentages s’échelonnent entre 2.3 % du
PIB pour le Chili et 10.7 % du PIB pour la Grèce. En 2021, les
quatrième et cinquième postes de dépenses ont été les services
généraux des administrations publiques et l’enseignement.
Les services généraux des administrations publiques (p. ex.
opérations concernant la dette publique, fonctionnement des
organes exécutifs et législatifs centraux et transferts entre
niveaux d’administration) ont représenté 5.4 % du PIB, tandis
que l’enseignement a représenté 5.1 % du PIB (tableau 11.4).
Entre 2019 et 2021, le poids dans le PIB des dépenses liées à la
protection sociale a augmenté de 2.4 points de pourcentage,
168
en moyenne de la zone OCDE. C’est au Chili (+8.5 p.p.) et aux
États-Unis (+4.5 p.p.) que l’augmentation a été la plus forte.
À l’inverse, c’est en Norvège que le poids des dépenses liées
à la protection sociale a le plus baissé (-0.9 p.p.). Sur la même
période, la deuxième catégorie de dépenses à progresser le plus
a été celle des affaires économiques, avec une hausse moyenne
de 1.7 p.p. de son poids dans le PTB sur l’ensemble de la zone
OCDE. C’est en Grèce que la hausse a été la plus marquée, avec
une progression de 6.9 p.p. (tableau 11.5).
Méthodologie et définitions
Les données relatives aux dépenses sont tirées des
Statistiques de l’OCDE sur les comptes nationaux (base
de données) et des statistiques d’Eurostat sur les
finances publiques (base de données), qui sont établies
conformément au Système de comptabilité nationale (SCN) ;
ce dernier constitue un ensemble de concepts, de
définitions, de nomenclatures et de règles approuvés au
plan international en matière de comptabilité nationale.
Le cadre du SCN 2008 a été mis en œuvre par tous les pays
de l’OCDE (voir l’annexe C). Les données sur les dépenses
sont ventilées selon la Classification des fonctions des
administrations publiques (CFAP, ou COFOG), qui classe
les dépenses publiques selon dix fonctions (niveau 1 de
la classification) : services généraux des administrations
publiques ; défense ; ordre et sécurité publics ; affaires
économiques ; protection de l’environnement ; logement
et équipements collectifs ; santé ; loisirs, culture et
culte ; enseignement ; et protection sociale. On trouvera
à l’annexe E des précisions sur la nature des dépenses
entrant dans chaque catégorie.
Pour en savoir plus
OCDE (2023), Perspectives économiques de l’OCDE, Rapport
intermédiaire mars 2023, Une reprise fragile, Éditions OCDE,
Paris, https://doi.org/10.1787/2d7536fc-fr.
Allen, R. (2022), « How budgeting systems can prepare better
for national emergencies: Six lessons from the COVID-19
crisis », OECD Journal on Budgeting, vol. 22, n° 1, https://doi.
org/10.1787/bdfca328-en.
Notes relatives aux graphiques
Les données sur le Chili, la Colombie et le Costa Rica ne sont pas prises
en compte dans la moyenne OCDE. On ne dispose pas de données
pour le Canada, le Mexique, la Nouvelle-Zélande et Türkiye. Les
données sur le Costa Rica et la Corée portent sur 2020, et non 2021.
G.6.2 et G.6.3 (Ventilation des dépenses des administrations publiques
par fonction en 2021, et évolution par rapport à 2019) sont disponibles
en ligne dans l’annexe G.
Panorama des administrations publiques 2023 © OCDE 2023
11. DÉPENSES PUBLIQUES
Ventilation des dépenses des administrations publiques par fonction (CFAP/COFOG)
11.4. StatLink 2 https://stat.link/i83tkb
11.5. StatLink 2 https://stat.link/2mh5ak
Panorama des administrations publiques 2023 © OCDE 2023
169
11. DÉPENSES PUBLIQUES
Ventilation des dépenses publiques liées aux fonctions de protection
sociale et de santé (CFAP/COFOG)
La pandémie de COVID-19 a montré l’importance de
systèmes de santé et de protection sociale robustes et agiles
pour préserver la vie et les conditions d’existence de la
population face aux crises. En moyenne de la zone OCDE,
la protection sociale et la santé constituent les principaux
postes de dépenses publiques, et leur poids s’est accru
pendant la pandémie. D’autres tendances démographiques
telles que l’allongement de l’espérance de vie et la faiblesse
des taux de fécondité ajoutent aux pressions financières qui
s’exercent sur les systèmes de santé et de protection sociale,
en accroissant la demande en faveur d’une offre de soins
plus étoffée et de meilleure qualité ainsi que la demande
de pensions de retraite et d’autres types d’aides et d’actions
sociales (OCDE, 2021).
La sous-catégorie des services de santé publique recouvre
les activités de recherche et de diffusion de l’information et
l’achat de vaccins et de masques pour la population. Même si
cette catégorie de dépenses est de taille relativement réduite
(0.6 % du PIB en 2021), son poids dans le total des dépenses de
santé a nettement augmenté entre 2019 et 2021 (+4.6 p.p. en
moyenne), sous l’effet de la pandémie de COVID-19. C’est en
Suisse (+18.5 p.p.), en Autriche (+11 p.p.) et en Hongrie (+ 10 p.p.)
que l’augmentation a été la plus marquée (tableau 11.7 et
tableau G.6.7 en ligne).
Au sein des dépenses de protection sociale, la sous-catégorie
la plus importante est celle des pensions de vieillesse, qui
représentait en 2021 10.9 % du PIB dans les pays de l’OCDE
également membres de l’UE. Ce sont l’Italie (14.3 %) et la
Finlande (13.9 %) qui ont consacré la plus forte proportion de
leur PIB aux pensions de vieillesse (tableau 11.6). Toutefois,
entre 2019 et 2021, le poids des dépenses de ce type au sein
des dépenses de protection sociale a baissé de 1.6 p.p. dans les
pays de l’OCDE membres de l’UE, en raison d’augmentations
relatives dans d’autres sous-catégories (tableau G.6.6 en ligne)
telles que les prestations de chômage, qui ont augmenté dans
une proportion équivalente sur cette période. En dehors de ce
groupe de pays, des reculs du poids des pensions de vieillesse
dans les dépenses de protection sociale ont été enregistrés
dans tous les pays pour lesquels des données sont disponibles.
Les reculs plus plus importants ayant été observés en Colombie
(-6.8 p.p.) et en Islande (-5.5 p.p.).
Les données relatives aux dépenses sont tirées des
Statistiques de l’OCDE sur les comptes nationaux (base
de données) et des statistiques d’Eurostat sur les
finances publiques (base de données), qui sont établies
conformément au Système de comptabilité nationale
(SCN) ; ce dernier constitue un ensemble de concepts, de
définitions, de nomenclatures et de règles approuvés au
plan international en matière de comptabilité nationale.
Le cadre du SCN 2008 a été mis en œuvre par tous les
pays de l’OCDE (voir l’annexe C). Les données relatives aux
dépenses sont ventilées, conformément à la Classification
des fonctions des administrations publiques (CFAP/
COFOG), entre dix grandes fonctions (de niveau 1). L’une de
ces grandes fonctions, la fonction « santé », est subdivisée
en 6 fonctions de niveau 2 : « produits, appareils et
matériels médicaux » ; « services ambulatoires » ; « services
hospitaliers » ; « services de santé publique » ; « recherchedéveloppement dans le domaine de la santé » ; et « santé
n.c.a. » (éléments non classés ailleurs). La fonction
« protection sociale » est subdivisée en 9 fonctions de
niveau 2 : « maladie et invalidité » ; « vieillesse » (c’està-dire pensions) ; « survivants » ; « famille et enfants » ;
« chômage » ; « logement » ; « exclusion sociale n.c.a. » ;
« recherche-développement dans le domaine de la
protection sociale » ; et « protection sociale n.c.a. ».
La deuxième sous-catégorie de dépenses de protection sociale
la plus importante est celle des prestations de maladie et
d’invalidité, qui représentait en 2021, en moyenne, 2.9 % du PIB
des pays de l’OCDE également membres de l’UE. La troisième
sous-catégorie la plus importante est celle des dépenses liées
aux familles et aux enfants, qui représentait, en moyenne,
1.9 % du PIB des pays de l’OCDE également membres de l’UE ;
c’est au Danemark que cette catégorie avait le poids le plus
élevé, avec 4.2 % du PIB (tableau 11.6).
Dans le domaine de la santé, la principale sous-catégorie de
dépenses reste celle des services hospitaliers, qui représentait
en 2021, en moyenne, 3.4 % du PIB des pays de l’OCDE
également membres de l’UE. Ces dépenses englobent des
dépenses d’investissement en équipements médicaux et
en installations médicales. Parmi les pays pour lesquels on
dispose de données, c’est au Royaume-Uni que les dépenses
liées aux services hospitaliers ont eu le poids le plus lourd,
avec 7.6 % du PIB (tableau 11.7). Dans les pays de l’OCDE
également membres de l’UE, le poids des services hospitaliers
dans le total des dépenses de santé a baissé, en moyenne, de
1.6 p.p. entre 2019 et 2021, poursuivant ainsi une tendance à
la baisse entamée en 2009 et qui s’explique peut-être par le
raccourcissement de la durée des hospitalisations constaté
au cours de la dernière décennie (OCDE, 2021). La deuxième
sous-catégorie de dépenses de santé la plus importante
est celle des services ambulatoires, qui représente, en
moyenne, 2.5 % du PIB. Cette catégorie, qui correspond à des
services assurés aussi bien à domicile que dans des locaux
de consultation, a vu son poids dans les dépenses de santé
baisser de 2.1 p.p. entre 2019 et 2021. C’est en Finlande que
le poids des dépenses ambulatoires a été le plus important
en 2021, avec 3.4 % du PIB.
170
Méthodologie et définitions
Pour en savoir plus
Bienassis, K., et al. (2023), « Advancing patient safety
governance in the COVID-19 response », Documents de travail
de l’OCDE sur la santé, n° 150, Éditions OCDE, Paris, https://
doi.org/10.1787/9b4a9484-en.
OCDE/Union européenne (2022), Health at a Glance: Europe
2022: State of Health in the EU Cycle, Éditions OCDE,
Paris, https://doi.org/10.1787/507433b0-en.
OCDE (2021), Panorama de la santé 2021 : Les indicateurs de l’OCDE,
Éditions OCDE, Paris, https://doi.org/10.1787/fea50730-fr.
Notes relatives aux graphiques
Pour plusieurs pays de l’OCDE non européens, on ne dispose pas de
données. Pour la Colombie et le Costa Rica, les données portent
sur 2020, et non 2021.
G.6.4 à G.6.7 (Ventilation des dépenses publiques liées aux fonctions
de protection sociale et de santé en 2021, et évolution depuis 2019)
sont disponibles en ligne dans l’annexe G.
Panorama des administrations publiques 2023 © OCDE 2023
11. DÉPENSES PUBLIQUES
Ventilation des dépenses publiques liées aux fonctions de protection sociale et de santé (CFAP/COFOG)
11.6. Ventilation des dépenses publiques liées aux fonctions de protection sociale, en pourcentage du PIB, 2021
Allemagne
Australie
Autriche
Belgique
Colombie
Costa Rica
Danemark
Espagne
Estonie
Finlande
France
Grèce
Hongrie
Irlande
Islande
Israël
Italie
Japon
Lettonie
Lituanie
Luxembourg
Norvège
Pays-Bas
Pologne
Portugal
République slovaque
République tchèque
Royaume-Uni
Slovénie
Suède
Suisse
OCDE-UE
Bulgarie
Croatie
Roumanie
Maladie
et invalidité
Vieillesse
Survivants
Famille et enfants
Chômage
Logement
Exclusion sociale
n.c.a.
R-D dans
le domaine de la
protection sociale
Protection sociale
n.c.a.
3.34
2.67
1.77
3.69
0.04
0.45
4.62
2.93
2.12
3.22
3.08
1.63
2.08
1.11
3.47
2.86
1.95
0.97
2.64
4.11
3.18
6.51
4.27
2.07
1.47
4.06
2.46
2.43
2.61
3.42
3.02
2.87
0.62
1.65
1.15
10.00
3.81
13.57
9.61
6.90
5.12
7.99
10.73
7.42
13.94
13.21
13.89
6.54
3.48
3.29
4.77
14.28
11.27
7.51
6.44
9.41
6.89
6.56
9.90
11.86
8.41
7.93
8.67
10.39
10.23
6.72
10.86
10.00
8.42
9.72
1.95
0.01
1.38
1.52
..
0.45
0.01
2.47
0.06
0.62
1.44
2.39
0.74
0.41
0.03
0.46
2.62
1.45
0.21
0.30
0.00
0.16
0.05
1.67
1.75
0.80
0.55
0.05
1.28
0.17
0.30
1.61
0.00
1.21
0.14
1.91
2.00
2.09
2.24
0.87
0.34
4.21
1.02
2.53
3.04
2.18
1.10
2.32
0.96
2.53
1.27
1.05
2.36
1.75
1.88
3.35
3.06
2.08
2.93
1.54
1.22
1.75
1.26
2.03
2.35
0.64
1.89
1.80
2.10
1.53
1.97
0.69
1.75
2.05
..
0.29
1.88
2.65
0.94
2.04
2.33
0.66
0.26
1.66
2.46
0.41
1.54
0.80
0.95
0.89
1.32
0.70
0.77
0.26
0.81
0.34
0.18
0.05
0.45
1.27
2.14
1.67
0.47
0.37
0.07
0.33
0.23
0.09
0.25
0.23
0.40
0.62
0.02
0.03
0.63
0.86
0.35
0.09
0.81
0.37
0.12
0.04
0.00
0.07
0.08
0.12
0.11
0.47
0.02
0.17
0.00
0.14
0.78
0.02
0.29
0.03
0.33
0.07
0.07
0.01
0.65
0.87
1.06
1.16
1.43
0.00
1.33
0.61
0.16
0.91
1.48
0.57
0.88
0.12
0.65
0.42
1.69
0.56
0.37
0.41
0.74
0.72
2.45
0.31
0.29
0.19
0.43
2.52
0.96
0.82
1.50
1.02
0.08
0.05
0.29
0.00
0.00
0.01
0.01
..
0.00
0.01
0.00
0.02
0.02
0.00
0.00
0.00
0.00
0.00
0.00
0.01
0.00
0.00
0.00
0.00
0.05
0.01
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.75
0.47
0.18
0.49
2.39
2.88
0.46
0.14
0.19
0.31
0.17
0.02
0.16
0.18
0.47
0.33
0.12
0.52
0.31
0.23
0.19
0.40
0.00
0.11
0.36
1.18
0.19
0.35
0.15
0.01
0.01
0.31
0.31
0.19
0.45
Sources : Statistiques de l’OCDE sur les comptes nationaux (base de données) ; statistiques d’Eurostat relatives aux finances publiques (base de données).
StatLink 2 https://stat.link/b4fp52
11.7. Ventilation des dépenses publiques liées aux fonctions de santé, en pourcentage du PIB, 2021
Australie
Autriche
Belgique
Colombie
Costa Rica
République tchèque
Danemark
Estonie
Finlande
France
Allemagne
Grèce
Hongrie
Islande
Irlande
Israël
Italie
Japon
Lettonie
Lituanie
Luxembourg
Pays-Bas
Norvège
Pologne
Portugal
République slovaque
Slovénie
Espagne
Suède
Suisse
Royaume-Uni
OCDE-UE
Bulgarie
Croatie
Roumanie
Produits, appareils
et matériels médicaux
Services ambulatoires
Services hospitaliers
Services de santé publique
R-D dans le domaine de
la santé
Santé n.c.a.
0.80
1.23
0.80
3.29
0.12
0.93
0.53
0.71
0.70
1.50
1.86
1.55
0.74
0.64
0.62
1.10
0.95
1.29
0.61
0.89
1.67
0.73
0.47
0.06
0.67
0.93
1.00
1.10
0.74
0.00
0.52
1.17
0.67
1.25
0.86
0.77
1.64
3.02
..
2.79
1.94
1.20
0.60
3.40
3.21
2.36
0.67
1.41
2.06
1.84
1.54
2.72
3.08
1.85
1.89
1.09
2.43
2.03
1.71
1.90
1.56
2.29
2.72
3.29
0.22
1.13
2.47
0.69
1.30
0.14
2.88
5.08
4.16
..
3.47
4.51
6.42
4.65
3.29
3.73
2.90
3.90
2.22
5.93
2.08
2.95
3.10
2.95
3.12
2.74
2.17
3.94
4.90
3.53
4.25
3.52
3.80
3.08
2.62
1.81
7.55
3.40
3.86
4.64
3.07
0.65
1.33
0.37
0.20
0.42
2.14
0.35
0.28
0.20
0.52
0.70
0.36
0.74
0.03
0.38
0.10
0.53
1.14
0.54
0.19
0.25
0.90
0.48
0.22
0.12
0.62
0.61
0.12
0.47
0.66
0.24
0.56
0.24
0.70
0.21
0.23
0.48
0.05
0.03
0.12
0.07
0.23
0.16
0.10
0.09
0.09
0.14
0.07
0.00
0.01
0.00
0.13
0.01
0.00
0.00
0.16
0.40
0.38
0.10
0.24
0.02
0.09
0.29
0.17
0.09
0.14
0.14
0.00
0.06
0.02
2.89
0.32
0.21
3.25
0.56
0.24
0.51
0.09
0.04
0.16
0.74
0.05
0.44
0.29
0.34
0.09
0.15
0.73
0.15
0.18
0.09
0.30
0.32
0.14
0.43
0.36
0.35
0.03
0.19
0.05
0.34
0.32
0.37
0.31
1.18
Sources : Statistiques de l’OCDE sur les comptes nationaux (base de données) ; statistiques d’Eurostat relatives aux finances publiques (base de données).
StatLink 2 https://stat.link/npm0d7
Panorama des administrations publiques 2023 © OCDE 2023
171
11. DÉPENSES PUBLIQUES
Rapport coût-efficacité
En termes économiques, l’efficacité mesure en quoi une activité
atteint les objectifs visés. Le rapport coût-efficacité, à savoir
le rapport entre un intrant et une réalisation intermédiaire
ou finale, rend compte du lien entre les ressources utilisées
et les résultats obtenus ; il s’agit d’un élément crucial pour
évaluer la réussite des politiques publiques. Dans les
secteurs de l’éducation et de la santé, on dispose d’éléments
de mesure des intrants et des réalisations suffisamment
développés et normalisés à l’échelle internationale pour
réaliser des évaluations comparatives pertinentes du rapport
coût-efficacité.
Santé
Les dépenses de santé représentent une part importante des
dépenses publiques, et leur poids devrait encore s’alourdir à
l’avenir, sous l’effet d’évolutions démographiques telles que
le vieillissement de la population (OCDE, 2021). Pour évaluer
le rapport coût-efficacité dans le domaine de la santé, on
compare les progrès réalisés sur le plan de l’espérance de vie
à la naissance (réalisation) au montant total des dépenses
de santé par habitant (intrant). Les dépenses courantes de
santé recouvrent des dépenses de santé publiques comme
privées ; ces dernières peuvent être particulièrement élevées
dans les pays qui, à l’instar des États-Unis, sont dépourvus
d’un vaste régime public de santé. L’espérance de vie n’est
pas un indicateur parfait de l’efficacité des dépenses de
santé, car elle est également influencée par des facteurs
extérieurs aux activités et dépenses médicales, y compris le
mode de vie, l’environnement physique et certains aspects
comportementaux. Il existe néanmoins une corrélation
positive entre les dépenses de santé et l’espérance de vie à la
naissance, avec des rendements décroissants (graphique 11.8).
Dans des pays tels que le Japon, la Corée et Israël, l’espérance
de vie est relativement élevée au regard du niveau de dépenses
de santé. En revanche, des pays tels que le Mexique, la Lettonie
et la Lituanie présentent une espérance de vie relativement
faible par rapport à d’autres pays engageant des dépenses
de santé similaires. S’agissant du Mexique, l’explication tient
peut-être en partie au niveau comparativement élevé des taux
d’obésité. Dans les deux pays baltes, l’abus de substances
psychoactives et le suicide ont nettement contribué à la
faiblesse de l’espérance de vie (Stumbrys et al., 2022). Les
États-Unis enregistrent l’une des espérances de vie les plus
faibles (77 ans), alors même qu’ils présentent, de loin, le plus
haut niveau de dépenses de santé par habitant de la zone
OCDE. Outre la question de l’accessibilité financière des soins,
des facteurs tels que les surdoses de produits stupéfiants, les
décès liés aux armes à feu et les troubles mentaux peuvent
expliquer ce score relativement bas (Ho, 2022).
Éducation
Tous les trois ans, dans le cadre de son Programme international
pour le suivi des acquis des élèves (PISA), l’OCDE évalue
les résultats des élèves de 15 ans en matière de lecture, de
mathématiques et de sciences. La comparaison entre les acquis
des élèves (évalués d’après les scores PISA) et les dépenses
éducatives cumulées par élève donne une mesure globale du
rapport coût-efficacité des systèmes éducatifs.
| 20,352
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https://openalex.org/W3106473506
|
OpenAlex
|
Open Science
|
CC-By
| 2,012
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Strange and identified hadron production at the LHC with ALICE
|
L. S. Barnby
|
English
|
Spoken
| 3,312
| 6,887
|
INTRODUCTION The aim of relativistic heavy-ion collision experiments is to detect and understand the
properties of the bulk QCD matter created in these collisions. Past experiments have
shown that the ability to perform measurements differentially with respect to the identity
of the final state hadrons is crucial to a full understanding of the evolution and dynam-
ics of the produced fireball [1, 2, 3]. Such measurements have also revealed anomalies
challenging the detailed modelling of the collision [4, 5]. The ALICE experiment was
designed with the goal of maximising the particle identification capability using tran-
sition and Cherenkov radiation detectors, calorimetry and, in particular for the analysis
presented here, identifying the most abundant species of charged hadrons over a wide
range of pT at mid-rapidity using dE/dx and time-of-flight techniques [6]. The excel-
lent tracking down to low pT also allows the reconstruction of weakly decaying neutral
strange particles via their charged decay modes. Strange and identified hadron production at the
LHC with ALICE
L. S. Barnby for the ALICE Collaboration L. S. Barnby for the ALICE Collaboration v:1110.4240v2 [nucl-ex] 20 Oct 2011 School of Physics and Astronomy, University of Birmingham, Birmingham. B15 2TT. UK Abstract. The ALICE detector was designed to identify hadrons over a wide range of transverse
momentum at mid-rapidity. Here measurements of light charged (π, K, p) and neutral (Λ, K0
S)
hadrons in Pb–Pb collisions at √sNN = 2.76 TeV are presented with additional data from a pp
reference at √s = 7 TeV. Such measurements are crucial for understanding the properties of
the fireball produced in heavy-ion collisions at the LHC. The particle-type dependence of the
spectra and the yields of particles extracted give information on the expansion dynamics and
chemical composition respectively. In addition studying the ratio of baryons to mesons may help in
understanding the mechanisms by which hadronisation takes place. We find that, when comparing
to data from √sNN = 200 GeV Au+Au collisions at RHIC, a more strongly expanding system is
created with a similar relative population of hadron species. We also see that collective effects or
complex mechanisms responsible for a relative enhancement of baryons have an influence at a much
higher pT than was previously seen. arXiv:1110.4240v2 [nucl-ex] 20 Oct Keywords: QGP, hadron production
PACS: 25.75-q,25.75.Dw,13.85.Hd Keywords: QGP, hadron production
PACS: 25.75-q,25.75.Dw,13.85.Hd EXPERIMENT The ALICE central barrel performs tracking of charged particles in a 0.5 T magnetic field
using a Time Projection Chamber (TPC) and Inner Tracking System (ITS). Particles
with large enough pT pass through the outer wall of the TPC and can go on to hit
a surrounding Time-of-Flight detector (TOF). Pb–Pb events were collected using a minimum bias trigger and several million events are used in this analysis. The Pb–Pb
data sample can be separated into centrality bins using the event-wise multiplicity in
the VZERO forward scintillator detectors in combination with a Monte Carlo Glauber
study [7]. Charged particle identification is achieved using two techniques. The specific
energy loss, dE/dx, can be calculated for each track from the ionisation in the TPC
gas (or ITS silicon) and compared to theoretical values from the Bethe-Bloch formula
which predicts the regions in momentum where π, K, and p signals can be separated. This separation between species can be used at low pT but near to the minimum of
dE/dx all three species are merged. In this range however the TOF can separate these
species so a combined pT spectrum can be extracted [8]. In this analysis the primary
yield of charged particles is reported; that is those emerging directly from the collision
or the decay of short-lived resonances and not the charged particles from the weak
decay of strange hadrons nor secondaries from the material. These are both excluded
using the distribution of the distance of closest approach to the primary interaction
vertex, which can be fitted to a template obtained from Monte Carlo events, where the
origin of the particle is known. The decay of neutral strange particles decaying into
charged daughters; Λ →pπ−and K0
S →π+π−, can be reconstructed and the invariant
mass distributions used for identification. The analysis follows the method used for
pp collisions but tighter cuts are made to further reduce the combinatorial background in
Pb–Pb events [9]. For both neutral and charged particle analyses the spectra are corrected
using efficiencies from Monte Carlo events having equivalent mean multiplicities. Charged Particles The combined pT spectra are obtained for each of eight centrality bins for π±, K±, p
and p and are shown, for positive particles only, in figure 1. The most noticeable features
are: the dramatic change in the shape of the spectra going from π through K to p; and
the shifting of the most probable values to higher pT, particularly for p but also for K. A
direct comparison of the most central spectra to Au–Au data at √sNN = 200 GeV is made
in figure 2. This shows how the spectra at LHC energy are much less steeply falling. A
first attempt at quantifying the changes using a parameterised blast wave function [10]
was made. The resulting fit parameters for the freeze-out temperature, Tfo, and mean
transverse velocity, β, are shown in figure 3 as 1-σ contours for each centrality class. Fits ranges 0.3–1.0 GeV/c, 0.2–1.5 GeV/c and 0.3–3.0 GeV/c were used for π, K p
respectively in order to avoid the region where a hard component of the spectum might
be expected and, at low pT, to avoid a strong contribution of resonances to π. There
appears to be a larger β, corresponding to stronger flow, than observed by STAR at
lower energy [11]. Tfo is very sensitive to the fit range so any change with respect to
RHIC needs further study. A blast wave was also fitted to each individual spectrum in
order to obtain pT-integrated yields, including the unmeasured part. These can be used
to form the ratios p/π and K/π for each centrality bin. The ratio p/π is almost constant
with centrality and is consistent with similar measurements in Au–Au collisions at RHIC
[12]. Charged Particles The ratio K/π shows a small rise from pp and peripheral collisions to the most (GeV/c)
T
p
0
0.5
1
1.5
2
2.5
3
(c/GeV)
T
dydp
N
2
d
-1
10
1
10
2
10
3
10
0-5%
5-10%
10-20%
20-30%
30-40%
40-50%
50-60%
60-70%
+
π
|y| < 0.5
2
syst
σ
+
2
stat
σ
(GeV/c)
T
p
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
2.2
(c/GeV)
T
dydp
N
2
d
-1
10
1
10
2
10
0-5%
5-10%
10-20%
20-30%
30-40%
40-50%
50-60%
60-70%
+
K
|y| < 0.5
2
syst
σ
+
2
stat
σ
(GeV/c)
T
p
0
0.5
1
1.5
2
2.5
3
(c/GeV)
T
dydp
N
2
d
-2
10
-1
10
1
10
2
10
0-5%
5-10%
10-20%
20-30%
30-40%
40-50%
50-60%
60-70%
p
|y| < 0.5
2
syst
σ
+
2
stat
σ
FIGURE 1. The centrality-selected pT spectra for identified π+(top) K+(middle) and p (bottom). Fits
are to a parameterised blast wave. (GeV/c)
T
p
0
0.5
1
1.5
2
2.5
3
(c/GeV)
T
dydp
N
2
d
-1
10
1
10
2
10
3
10
0-5%
5-10%
10-20%
20-30%
30-40%
40-50%
50-60%
60-70%
+
π
|y| < 0.5
2
syst
σ
+
2
stat
σ
(GeV/c)
T
p
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
2.2
(c/GeV)
T
dydp
N
2
d
-1
10
1
10
2
10
0-5%
5-10%
10-20%
20-30%
30-40%
40-50%
50-60%
60-70%
+
K
|y| < 0.5
2
syst
σ
+
2
stat
σ (GeV/c)
T
p
0
0.5
1
1.5
2
2.5
3
(c/GeV)
T
dydp
N
2
d
-1
10
1
10
2
10
3
10
0-5%
5-10%
10-20%
20-30%
30-40%
40-50%
50-60%
60-70%
+
π
|y| < 0.5
2
syst
σ
+
2
stat
σ (GeV/c)
T
p
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
2.2
(c/GeV)
T
dydp
N
2
d
-1
10
1
10
2
10
0-5%
5-10%
10-20%
20-30%
30-40%
40-50%
50-60%
60-70%
+
K
|y| < 0.5
2
syst
σ
+
2
stat
σ (GeV/c)
T
p
0
0.5
1
1.5
2
2.5
3
(c/GeV)
T
dydp
N
2
d
-2
10
-1
10
1
10
2
10
0-5%
5-10%
10-20%
20-30%
30-40%
40-50%
50-60%
60-70%
p
|y| < 0.5
2
syst
σ
+
2
stat
σ FIGURE 1. Charged Particles The centrality-selected pT spectra for identified π+(top) K+(middle) and p (bottom). Fits
are to a parameterised blast wave. central collisions and is also consistent with previous lower energy data [13]. central collisions and is also consistent with previous lower energy data [13]. (GeV/c)
T
p
0
0.5
1
1.5
2
2.5
3
-1
dy) (GeV/c)
T
N/(dp
2
d
-1
10
1
10
2
10
3
10
0-5% most central
ALICE Preliminary
= 2.76 TeV
NN
s
ALICE, Pb-Pb,
= 200 GeV
NN
s
STAR, Au-Au,
= 200 GeV
NN
s
PHENIX, Au-Au,
S
0
K
-π
-
K p
FIGURE 2. The pT spectra for π−, K0
S, K−, and p for the most central Pb–Pb (0-5%) collisions (solid
markers) plotted with those measured in √sNN = 200 GeVAu–Au collisions (open symbols.) (GeV/c)
T
p
0
0.5
1
1.5
2
2.5
3
-1
dy) (GeV/c)
T
N/(dp
2
d
-1
10
1
10
2
10
3
10
0-5% most central
ALICE Preliminary
= 2.76 TeV
NN
s
ALICE, Pb-Pb,
= 200 GeV
NN
s
STAR, Au-Au,
= 200 GeV
NN
s
PHENIX, Au-Au,
S
0
K
-π
-
K p FIGURE 2. The pT spectra for π−, K0
S, K−, and p for the most central Pb–Pb (0-5%) collisions (solid
markers) plotted with those measured in √sNN = 200 GeVAu–Au collisions (open symbols.) > (c)
β
<
0
0.2
0.4
0.6
(GeV)
fo
T
0
0.05
0.1
0.15
0.2
, K, p
π
Au-Au, pp, 0.2 TeV,
, K, p
π
Pb-Pb, 2.76 TeV,
Preliminary
FIGURE 3. 1-σ contours in the T-β plane from a simultaneous fit of a parameterised blast wave
function to the π±, K, and p pT spectra for various centrality classes. Pb–Pb collisions from the ALICE
experiment in red, Au–Au collisions from the STAR experiment in blue. Most central data lie to the right. FIGURE 3. 1-σ contours in the T-β plane from a simultaneous fit of a parameterised blast wave
function to the π±, K, and p pT spectra for various centrality classes. Pb–Pb collisions from the ALICE
experiment in red, Au–Au collisions from the STAR experiment in blue. Most central data lie to the right. Neutral Particles The pT spectra of Λ and K0
S have also been extracted for each centrality bin. As the
systematic uncertainties on the efficiency correction are still under study the preliminary
spectra themselves are not yet ready. However the study reveals that the ratio of the
efficiencies for each particle, as a function of pT , is rather stable with respect to changing
the centrality of the collision. In particular in the pT range 2.5-5.5 GeV/c the variation of
the efficiency ratio between the most central and the most peripheral centrality selections
is below 2%. This allows the Λ/ K0
S ratio to be calculated with an estimated systematic
uncertainty of 10% and the resulting curves for each centrality are shown in figure 4 (upper). Also shown are the ratios in pp collisions at √s = 0.9 and 7 TeV [9]. The pp data
demonstrate that in the TeV range the maximum value of the ratio is almost constant
and it is reasonable to assume that pp collisions at √s = 2.76 TeV would show the same
maximum. Taking this pp baseline the ratio is observed to have a maximum which rises
strongly going to peripheral and then to central events, with a total increase up to a factor
of three. The value of pT at which the maximum is reached is also increasing by several
hundred MeV/c. The data are compared to a similar measurement previously made by
STAR in figure 4 (lower) [14]. To facilitate the comparison the lower energy data were
scaled by the Λ/Λ ratio measured for each centrality [15], assuming it is constant in pT,
because it has previously been noted that there is a √s-dependence of the ratio [16],
presumably due to the change in the baryo-chemical potential. The ALICE data were
not scaled in this way because the anti-baryon/baryon ratio in LHC collisions is very
close to one [17]. The ratio in peripheral 60-80% collisions is very similar in shape for
the two collision systems with only a small change in the magnitude. In the most central
0-10% however the shape is quite different with the enhancement of the Λ extending to
a much larger pT in the higher energy data. This is qualitatively in agreement with some
predictions [18]. CONCLUSIONS Pb–Pb collisions at √sNN = 2.76 TeV reveal a number of similarities to Au–Au collisions
at RHIC; the ratios of the yields are the same within the experimental uncertainties,
the spectra are compatible with a strong collective motion which increases going to
more central collisions and there is a growth of the Λ/ K0
S ratio in the pT region 2-4
GeV/c, also with centrality. There are however some notable differences; the pT spectra
are much flatter giving a transverse flow velocity in a blast wave parameterisation 10%
larger than that in √sNN = 200 GeVAu–Au collisions and the enhanced baryon-to-meson
ratio extends to a pT of around 6 GeV/c. This may imply that the influence of particles
participating in the collective dynamics of the system extends to a higher pT than has
previously been observed. y
8.
K. Aamodt, et al., Physics Letters B 693, 53–68 (2010). 7.
K. Aamodt, et al., Phys. Rev. Lett. 106, 032301
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8. K. Aamodt, et al., Physics Letters B 693, 53–68 (2010). (GeV/c)
T
p
0
1
2
3
4
5
6
7
8
S
0
/ K
Λ
0
0.5
1
1.5
2
2.5
0-5 % centrality
20-40 % centrality
40-60 % centrality
60-80 % centrality
80-90 % centrality
= 7 TeV)
s
(pp at
= 0.9 TeV)
s
(pp at
data points include stat. errors
estimated syst. error ~10 %
Preliminary
= 2.76 TeV, |y|<0.75
NN
s
Pb-Pb at
(GeV/c)
T
p
0
1
2
3
4
5
6
7
8
0
S
/ K
Λ
0
0.5
1
1.5
2
2.5
3
= 2.76 TeV, |y|<0.75
NN
s
Pb-Pb at
data points include stat. errors
estimated syst. error ~10 %
Λ
/
Λ
×
STAR: Au-Au 200 GeV
ALICE:
with 10% feed-down correction
0-5% centrality
60-80% centrality
0-5% centrality
60-80% centrality
Preliminary
FIGURE 4. (Upper panel.) The ratio of Λ to K0
S as a function of pT for five centrality classes in Pb–Pb
collisions. 18. R. Fries, and B. Müller, The European Physical Journal C - Particles and Fields 34, s279–s285
(2004), URL http://dx.doi.org/10.1140/epjcd/s2004-04-026-6. 17. K. Aamodt, et al., Phys. Rev. Lett. 105, 072002 (2010). 16. M. M. Aggarwal, et al., Phys. Rev. C 83, 024901 (2011). 32, S105 (2006), URL http://stacks.iop.org/0954 3899/32/i 12/a S13.
15. J. Adams, et al., Phys. Rev. Lett. 98, 062301 (2007), URL http://link.aps.org/doi/10.
1103/PhysRevLett.98.062301. REFERENCES Also shown the same ratio in pp collisions at two energies. (Lower panel.) A comparison
between the ratio measured by ALICE (solid markers) and STAR (open symbols) for selected centralities. (GeV/c)
T
p
0
1
2
3
4
5
6
7
8
S
0
/ K
Λ
0
0.5
1
1.5
2
2.5
0-5 % centrality
20-40 % centrality
40-60 % centrality
60-80 % centrality
80-90 % centrality
= 7 TeV)
s
(pp at
= 0.9 TeV)
s
(pp at
data points include stat. errors
estimated syst. error ~10 %
Preliminary
= 2.76 TeV, |y|<0.75
NN
s
Pb-Pb at (GeV/c)
T
p
0
1
2
3
4
5
6
7
8
0
0.5
1
(GeV/c)
T
p
0
1
2
3
4
5
6
7
8
0
S
/ K
Λ
0
0.5
1
1.5
2
2.5
3
= 2.76 TeV, |y|<0.75
NN
s
Pb-Pb at
data points include stat. errors
estimated syst. error ~10 %
Λ
/
Λ
×
STAR: Au-Au 200 GeV
ALICE:
with 10% feed-down correction
0-5% centrality
60-80% centrality
0-5% centrality
60-80% centrality
Preliminary
FIGURE 4. (Upper panel.) The ratio of Λ to K0
S as a function of pT for five centrality classes in Pb–Pb
collisions Also shown the same ratio in pp collisions at two energies (Lower panel ) A comparison (GeV/c)
T
p
0
1
2
3
4
5
6
7
8
0
S
/ K
Λ
0
0.5
1
1.5
2
2.5
3
= 2.76 TeV, |y|<0.75
NN
s
Pb-Pb at
data points include stat. errors
estimated syst. error ~10 %
Λ
/
Λ
×
STAR: Au-Au 200 GeV
ALICE:
with 10% feed-down correction
0-5% centrality
60-80% centrality
0-5% centrality
60-80% centrality
Preliminary (GeV/c)
T
p
0
1
2
3
4
5
6
7
8
0
S
/ K
Λ
0
0.5
1
1.5
2
2.5
3
= 2.76 TeV, |y|<0.75
NN
s
Pb-Pb at
data points include stat. errors
estimated syst. error ~10 %
Λ
/
Λ
×
STAR: Au-Au 200 GeV
ALICE:
with 10% feed-down correction
0-5% centrality
60-80% centrality
0-5% centrality
60-80% centrality
Preliminary FIGURE 4. (Upper panel.) The ratio of Λ to K0
S as a function of pT for five centrality classes in Pb–Pb
collisions. Also shown the same ratio in pp collisions at two energies. (Lower panel.) A comparison
between the ratio measured by ALICE (solid markers) and STAR (open symbols) for selected centralities. 0. E. REFERENCES Schnedermann, J. Sollfrank, and U. Heinz, Phys. Rev. C 48, 2462–2475 (1993), URL ht
//link.aps.org/doi/10.1103/PhysRevC.48.2462. 11. J. Adams,
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B. I. Abelev, et al., Phys. Rev. C 79, 034909 (2009), URL http://link.aps.org/doi/10
1103/PhysRevC.79.034909. 14. M. A. C. Lamont, and the STAR Collaboration, Journal of Physics G: Nuclear and Particle Physics
32, S105 (2006), URL http://stacks.iop.org/0954-3899/32/i=12/a=S13. 32, S105 (2006), URL http://stacks.iop.org/0954 3899/32/i 12/a S13. 15. J. Adams, et al., Phys. Rev. Lett. 98, 062301 (2007), URL http://link.aps.org/doi/10. 1103/PhysRevLett.98.062301. gg
y
17. K. Aamodt, et al., Phys. Rev. Lett. 105, 072002 (2010). ,
,
y
,
(
)
18. R. Fries, and B. Müller, The European Physical Journal C - Particles and Fields 34, s279–s285
(2004), URL http://dx.doi.org/10.1140/epjcd/s2004-04-026-6.
| 21,829
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Pour une anthropologie matérialiste de l’homme néolibéral : gouvernementalité ou domination ?
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Spoken
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Et on pense par avance ce qui n’existe pas encore. Donc, prendre acte par avance, qu’est-ce d’autre qu’annoncer comme inéluctable une évolution historique? Ce qui revient à la présenter comme souhaitable. En l’occurrence, ici, c’est annoncer comme horizon incontournable (et donc souhaitable en raison) la fin des nations et des pouvoirs des Étatsnations, au profit d’un marché mondialisé (donc des puissances d’argent qui s’y épanouissent). Annoncer une telle inéluctable fin n’est-ce pas tout simplement mener la bataille des idées pour faire basculer le monde en servant d’auxiliaires aux puissances d’argent qui orchestrent la mondialisation? Dans un tel paysage politique et théorique, on ne s’étonne pas que cette discussion entre Jacques Donzelot et Colin Gordon s’achève par une commune célébration de « l’Europe », celle de la concurrence libre et non faussée. On est aux antipodes de la critique du néolibéralisme que certains veulent lire dans le cours de 1978-79 de Foucault.
II-3) L’État pénal
Il est une tout autre “entrée” dans l’analyse du néolibéralisme que celle de la gouvernementalité et de son histoire (ce que nous venons de voir en empruntant les voies ouvertes par Foucault) ; c’est celle proposée par la sociologie de la domination de Bourdieu et de ses collaborateurs. Parmi ces derniers, Loïc Wacquant, qui avait cosigné avec Bourdieu, en 208. Ibid., p.93. 151 2000, un article publié dans Le Monde diplomatique. Le cœur de cet article tient dans ces deux longues phrases : « [...] ces lieux communs que le ressassement médiatique transforme en sens commun universel parviennent à faire oublier qu’ils ne font bien souvent qu’exprimer, sous une forme tronquée et méconnaissable, y compris pour ceux qui les propagent, les réalités complexes et contestées d’une société historique particulière, tacitement constituée en modèle et en mesure de toutes choses : la société américaine de l’ère postfordiste et postkeynésienne. Cet unique superpouvoir, cette Mecque symbolique de la Terre est caractérisée par le démantèlement délibéré de l’État social et l’hypercroissance corrélative de l’État pénal, l’écrasement du mouvement syndical et la dictature de la conception de l’entreprise fondée sur la seule « valeur-actionnaire », et leurs conséquences sociologiques, la généralisation du salariat précaire et de l’insécurité sociale, constituée en moteur privilégié de l’activité économique. »209 On y retrouve, de manière condensée les points saillants de la critique bourdieusienne du néolibéralisme210 (dans le cas de Bourdieu, à l’inverse des ambiguïtés de Foucault, l’abord du néolibéralisme est expressément critique et combatif) : on part du constat que le “modèle” néolibéral – alors présenté à l’opinion comme la promesse enchantée d’une sortie radieuse du vieux monde sclérosé de l’État-providence et de son welfare211 – est une marchandise idéologique d’importation qui nous vient des États-Unis et du reaganisme. Cette marchandise bénéficie en France – comme en Europe en général – d’une fascination qui se traduit par un « ressassement médiatique » qui sature le sens commun212. Aux USA
en Europe, il 209. Pierre Bourdieu et Loïc Wacquant, « La nouvelle vulgate planétaire », Le Monde diplomatique, mai 2000, p. 6. 210. Points saillants qu’elle partage avec les critiques portées par l’altermondialisme et la gauche de gauche (que Bourdieu n’a jamais prétendu réduire à la configuration de la gauche de la gauche). 211. Pensons, par exemple, à l’émission de “pédagogie” diffusée par la chaîne de télévision TF1 (alors pas encore privatisée) et concoctée par Yves Montand et Laurent Joffrin sous le slogan « Vive la crise! » (voir Frédéric Lebaron, « Genèses du néolibéralisme. Contribution à une analyse sociologique du cas français », mai 2014 (conférence prononcée en 1998), en ligne : http://www.grand-angle-libertaire.net/geneses-duneoliberalisme-par-frederic-lebaron/). 212. La très forte concentration de la propriété des médias français dans les mains des quelques puissances d’argent qui sont les bénéficiaires immédiats de la mondialisation libérale cause le pesant envoûtement journalistique pour la « mondialisation heureuse » des années 1990-2000 (devenue la mondialisation sans retour des années 2000-2010). Le « cercle de la raison » d’Alain Minc prend au XXIe siècle la forme du spectre de la « dette » léguée à nos enfants et petits-enfants. La médiocrité scientifique de ce discours ne peut faire illusion qu’à la condition de le soustraire au souci du pluralisme de l’information économique et sociale (la rubrique Social a disparu depuis de longues années d’un quotidien — dit « de référence » — comme Le Monde) : on a donc sur les ondes, les plateaux TV et dans les colonnes de la presse écrite, un monopole quasi exclusif du discours économique orthodoxe (un discours qui se réduit chez les experts du commentaire politique – la vingtaine d’indéboulonnables éditorialistes qui quadrillent l’intégralité des médias – à quelques slogans rabâchés jusqu’à plus soif). Et, quand il n’a plus la patience de s’appuyer sur de sommaires montages de chiffres et de données, ce discours recourt à son argument-massue privilégié : critiquer le libre-échangisme et l’extension universelle de la concurrence, c’est être un souverainiste nostalgique de l’État hypertrophié (étant admis comme une vérité 152 existe de nombreux Think Tanks qui alimentent et diffusent ce catéchisme justificateur – pour ne pas dire apologétique – de la mondialisation libérale, de sa nécessité historique et de ses vertus salvatrices213. Cette diffusion d’argumentaires préconstruits et uniformes occulte systématiquement tous les éléments qui pourraient les nuancer ou les invalider Elle s’opère de préférence, non auprès du grand public, mais auprès des décideurs (entrepreneurs, experts universitaires, journalistes dominants, communicants, managers, intellectuels à gages – la plupart cumulant individuellement plusieurs de ces “compétences” convertibles l’une dans l’autre) avec pour objectif de contrôler idéologiquement les lieux de pouvoir, d’influence et, surtout, de décision. La stratégie de ces Think Tanks – qui bénéficient des moyens confortables qui leur sont généreusement alloués par de grosses entreprises et, aussi, par des subventions publiques de l’Union européenne ou de certains États – réitère celle qui a si bien réussi à la Société du Mont-Pèlerin quand il s’est agi de renverser l’État-Providence et d’expulser progressivement le keynésianisme des sphères gouvernementales. Pour le grand public, on se contentera de slogans caricaturaux et épais pour stigmatiser la « mamma étatique » – expression utilisée par Claude Imbert (défunt éditorialiste du Point), qui la servait « avec une constance proche du radotage » pour « dénoncer l’intervention d’un État bridant, avec sa cohorte de fonctionnaires assistés, les merveilleuses initiatives des “entrepreneurs” privés »214. On le voit : quand il s’agit de vanter les bienfaits d’une mondialisation irréversible, la réflexion néolibérale se charge d’une (apparente) complexité savante dont elle se dépouille subitement dès qu’il s’agit de matraquer les esprits avec des clichés dont la grossièreté fait frémir. Dès lors qu’il s’agit de marteler les slogans du catéchisme néolibéral (trop d’État, trop de dépenses publiques, trop de dette publique, trop de fonctionnaires, trop de taxation du capital, etc.), on dissimule la propagande sous les oripeaux de la « pédagogie ». Il faut fabriquer le consensus, rééduquer les envoûtés du “populisme”, embrigader les esprits frustes qui ne peuvent accéder aux démonstrations élégantes du discours néolibéral. Les porte-drapeaux de la dénonciation du “populisme” et de la défense d’évidence que l’État et ses fonctionnaires sont forcément archaïques, gaspilleurs et potentiellement totalitaires) ; se livrer à une telle critique, c’est donc être un pitoyable xénophobe, un inquiétant populiste (fasciné, de fait, par un protectionnisme odieux qui n’est rien d’autre que la version contemporaine du fascisme). Presse “de gauche” et presse de droite, médias « populaires » et médias « cultivés » peuvent benoîtement communier dans ces clichés et nous les resservir quotidiennement
213. À ce propos, voir Antoine Schwartz, « Les think tanks et la consolidation d'une vision économique du social », Informations sociales 2010/1 (n° 157), p. 60-68. 214. Serge Halimi, Le Grand Bond en arrière, Fayard, Paris, 2004, p. 30, note 1.
Cette note se poursuit en relevant un paradoxe savoureux : « La règle ne semble pas cependant s’appliquer à François Pinault, propriétaire du Point, qui fut largement comblé par la “mamma” lorsqu’il racheta à vil prix des actions du Crédit Lyonnais, alors nationalisé. » 153 des élites se recrutent prioritairement dans les rangs des quelques éditorialistes et fast thinkers qui flétrissent de l’adjectif “populiste” toute critique de la globalisation et qui célèbrent une “complexité” inaccessible aux catégories socio-professionnelles populaires (des catégories composées d’individus frustes et menaçants qui seraient intoxiqués par quelques agitateurs aveuglés par un anti-libéralisme dogmatique). Les contempteurs du “populisme” brandissent la “complexité” comme un brevet de subtilité et de respectabilité ; mais ils ne semblent jamais disposer du temps nécessaire pour déployer la complexité sous la forme d’une argumentation détaillée (le temps indispensable aux méandres de l’esprit de finesse n’entre plus dans le format contraint du travail médiatique contemporain). Chiens de garde et nouveaux chiens de garde travaillent en meute ; en aboyant selon des mélodies qui ne sont guère variées ou nuancées. On trouve ensuite dans l’article cosigné par Bourdieu et Loïc Wacquant en 2000 les caractéristiques essentielles des politiques néolibérales : l’écrasement du mouvement syndical (Thatcher, une fois parvenue au pouvoir, avait entamé son œuvre politique par une lutte à mort contre le syndicat britannique des mineurs) ; la dictature exercée par la finance sur les entreprises (l’actionnaire a pris le pouvoir sur l’industriel, et il engloutit des dividendes sans cesse de plus en plus importants) ; la précarisation toujours grandissante du travail et la généralisation d’une insécurité sociale chaque jour plus menaçante. En quelque sorte, la lente agonie d’un État-providence attaqué de tous côtés. Il y a cependant, une autre caractéristique qui a été mentionnée d’emblée parce qu’elle est encore plus importante ; en effet, elle est plus déterminante que les constats précédents parce qu’elle est l’axe central de l’analyse des politiques néolibérales par la sociologie de Bourdieu et c’est celle qui résume et condense toutes les autres : le néolibéralisme modifie l’équilibre entre l’État social et l’État pénal, il contracte l’État social et, corrélativement, gonfle l’État pénal par un effet de vases communicants. Avec le néolibéralisme, l’État se coupe la main gauche (celle qui redistribue) et hypertrophie sa main droite (celle qui réprime). L’incarcération de masse aux États-Unis
C’est bien des États-Unis que nous vient cette évolution. Elle a été analysée par Loïc Wacquant. Et l’étude des données chiffrées de l’emprisonnement aux USA, dans le dernier quart du XXe siècle, est particulièrement éloquente : « Entre 1975 et 2000, les États-Unis ont multiplié par cinq leur population sous écrou pour devenir le leader mondial de l’incarcération avec 2 millions de détenus – 154 chose que j’ignorais alors et dont je ne tenais aucun compte analytique, comme tout sociologue travaillant sur race et classe en Amérique (le premier qui l’a fait est un juriste, Michael Tonry, dans Malign Neglect, un livre-clef paru en 1995, qui a attiré mon attention parce que je voulais utiliser ce titre pour un de mes ouvrages). »215 On peut préciser et actualiser les chiffres impressionnants données par Loïc Wacquant, en 2008, au sujet de la croissance exponentielle de l’État pénal aux USA : en 1971, il y avait 200 000 prisonniers ; en 2014, on en dénombre 2,4 millions. Et ceci sans compter les près de 5 millions d’individus placés sous contrôle judiciaire. Les États-Unis représentent aujourd’hui environ 5 % de la population mondiale, mais ils constituent environ 25 % de la population carcérale du monde. En 2008 l’Allemagne comptait 93 détenus pour 100 000 habitants et les USA 750.216 Dans la population incarcérée, la surreprésentation des Afro-américains est plus que frappante. Cette surreprésentation a un peu diminué durant les années 2000 ; mais c’est parce que la population hispanique a, à son tour, connu un développement spectaculaire de son taux de condamnations et d’enfermements, avec une prolifération des sanctions pénales. Cette politique de démultiplication de l’emprisonnement a connu des étapes successives : c’est la présidence Nixon qui enclenche le processus en déclarant la « guerre à la drogue ». Cette guerre justifie l’augmentation des budgets de la sécurité (police et prisons). La présidence Reagan poursuit et aggrave cette politique. Cette présidence fait des choix budgétaires très significatifs : le taux d’imposition des très grosses fortunes baisse vertigineusement alors que les dépenses sécuritaires augmentent énormément. Bien entendu, cela signifie concrètement que l’on taille dans les budgets sociaux. L’État pénal remplace spectaculairement l’État social ; la fin de l’État-providence, c’est le gonflement de l’État qui punit et enferme. La présidence Clinton ne modifie pas cette tendance, au contraire elle l’aggrave : en 1994, Clinton fait voter le Violent Crime Act Contrôle and Law Enforcement Act : un budget de 30 milliards de dollars, une création de 100 000 postes de policiers, la peine de mort étendue à 60 nouveaux crimes, la construction de nouvelles prisons, la généralisation de la « loi des trois coups » (qui oblige les juges à prononcer une peine d’emprisonnement perpétuel pour la troisième condamnation217). Et, parallèlement, Clinton « sonne la fin du régime de protection sociale »
.218 Sous la présidence Bush, le Patriot Act 215. Loïc Wacquant, « Le corps, le ghetto et l’État pénal. Entretien réalisé par Susana Durao », Labyrinthe n° 31, 2008, texte non paginé. 216. Voir Keeanga-Yamahtta Taylor, Black Lives Matter. Le renouveau de la révolte noire américaine, trad. C. Izoard, Agone, Marseille, 2017, p. 204 et Michelle Alexander, Incarcération de masse et nouvelle
ségrég
ation
raciale
aux
États-Unis
, trad.
A. Scherrer, Syllepse, Paris, 2017,
p
.
17. 217
. La Présidence Trump a supprimé la loi des trois coups en décembre 2018. 218. Pour tout cela, voir Keeanga-Yamahtta Taylor
, op. cit., pp. 206-207. 155 autorise l’Armée à transférer ses surplus à la police. En quelques années, des centaines de milliers de dollars de matériels militaires deviennent ainsi la propriété de la police. Cela augmente significativement la puissance de l’armement de la police et « transforme en soldats les policiers des ghettos des centres-villes »219. On peut parler d’une militarisation de la police (donc on ne peut guère ensuite s’étonner que la police des États-Unis tue nettement plus que n’importe quelle autre police de pays développés). On assiste aussi à une transformation de la législation : alors que la délinquance est régulièrement en baisse depuis 1991, on rend condamnables les « incivilités » et les atteintes à la « qualité de la vie ». Ainsi, s’asseoir sur un trottoir est aujourd’hui un délit dans plus de la moitié des villes américaines ; 9 % de ces mêmes villes interdisent de donner ou de partager de la nourriture avec des sans-abris. Bien entendu, « la multiplication de ces infractions est allée de pair avec le sabrage ou l’élimination des emplois ou des aides aux travailleurs pauvres ». On est toujours dans la même logique : moins de social, plus de pénal. Cette inflexion de la législation a un effet non négligeable ; en effet, condamnations et amendes se multiplient et les municipalités américaines se financent de plus en plus grâce à cette ressource : « Du fait de la frilosité des politiciens à augmenter les impôts des contribuables aisés ou des grandes entreprises, les municipalités du XXIe siècle sont en effet de plus en plus dépendantes financièrement des contraventions dressées pendant ou après les contrôles : les policiers jouent un rôle croissant dans la collecte des revenus municipaux »220 Ainsi, on en arrive à un paradoxe inquiétant : les grosses villes américaines réduisent les budgets sociaux (voire ferment des écoles comme à Chicago en 2013), mais paient des sommes colossales pour mettre un terme aux poursuites auxquelles elles doivent faire face à propos des nombreuses violences commises par leurs policiers. Toute cette évolution multiplie les pauvres et, simultanément, elle les criminalise. Mais cela ne suffit pas à l’État pénal : il faut encore faire payer les pauvres. Les amendes et les frais de justice se multiplient. Et, s’ils ne sont pas payés, ils se démultiplient. Ce qui peut aboutir à une nouvelle condamnation. Et cette condamnation va s’accompagner de nouvelles amendes et de nouveaux frais. « En 2004, selon les chiffres du Département de la Justice, 66 % des détenus étaient “redevables de frais de justice, d’indemnités, d’amendes et de frais divers” – contre 21 % en 1991. »221 L’État pénal emprisonne de la sorte le pauvre dans un 219. Ibid., p. 208. 220. Ibid., p. 217. 221. Ibid., p. 219. 156 endettement insurmontable dont il ne peut jamais sortir, ce qui le conduit à de nouvelles condamnations, ce qui l’expose à de nouveaux emprisonnements ; et ainsi de suite à l’infini. Où l’on retrouve le mécanisme de la dette comme un dispositif caractéristique de l’assujettissement dans les sociétés néolibérales. Mais sous une forme inattendue. Il existe encore deux aspects notables de cette extension impressionnante de l’État pénal : la pénalisation de la pathologie psychique et le traitement réservé aux enfants. Ces deux aspects conduisent à une interrogation simple : la stratégie d’enfermement qui s’est développée aux USA ne correspond-elle pas, jusque dans ses cibles, à ce que Foucault décrivait comme des dispositifs disciplinaires? En effet, il va de soi que la première cible de cette stratégie, c’est le pauvre, c’est-à-dire celui qui, a priori, est le plus susceptible de faire gripper le mécanisme de la reproduction de la domination (non par choix, mais par effet de structure). De plus, les fous et les enfants sont également des cibles de l’État pénal américain. Commençons par ce qui concerne la pathologie psychique : « les prisons d’État sont devenues la principale destination des malades mentaux auteurs de délits. »222 Cela s’explique de manière fort simple : la compression des budgets sociaux (il faut bien trouver les moyens pour financer en priorité la police et les prisons) conduit les villes américaines à réduire drastiquement le nombre des lieux d’accueil pour les pathologies psychiques. Donc, quand un individu souffrant de troubles psychiques commet un délit, sa “prise en charge” ne peut plus s’effectuer dans un service spécialisé. Il est condamné pour le délit commis et enfermé dans une prison. Ainsi, en 2017, 17 % des détenus américains sont considérés comme atteints de maladie mentale. Plus impressionnant encore : « au moins la moitié des personnes tuées par la police depuis 2000 étaient atteintes d’une forme ou d’une autre de maladie mentale. »223 On s’aperçoit donc que, sur le territoire américain, là où l’art néolibéral de gouverner trouve son expression la plus large, sous le régime de l’État pénal, on enferme les fous, mais dans des prisons, avec les délinquants de toutes sortes. C’est le crime ou le délit qui prime sur la pathologie mentale. Ce qui change aussi sans doute profondément par rapport aux dispositifs disciplinaires analysés par Foucault, c’est que la volonté de dresser les corps a probablement largement disparu. Le renoncement à l’ambition disciplinaire explique sans doute qu’un nombre significatif de supposés délinquants atteints de pathologies mentales manifestes soient, de fait, physiquement éliminés (pour rappel : comparée aux polices des autres pays développés, la police américaine est celle qui tue le plus). On n’est pas dans une logique 222. Ibid., p.
. 223. Ibid., p. 213. 157
d’extermination, mais on ne peut pas non plus considérer ces chiffres et les évolutions qu’ils indiquent comme dénués de signification. Et cette évolution trouve probablement son explication dans un changement assez simple à constater : pour qu’il existe des dispositifs disciplinaires, il faut une bonne raison pour discipliner. Cette raison, c’est que la population doit être “employable”, c’est-à-dire qu’elle puisse être mise au travail pour être productive. Or, à l’ère des sociétés néolibérales, une partie significative de la population, de ce point de vue, ne présente plus guère d’intérêt : l’économie (en tout cas l’économie officielle) n’a plus besoin des bras de tous ces travailleurs potentiels. Une partie de la population devient donc inutile, comme en surplus, de trop : « La logique de la “société” civile produit inévitablement une classe croissante d’individus qui ne sont pas seulement menacés de pauvreté, ou d’injustice, mais qui sont tout simplement “de trop”. »224 Que faire du surplus? Il se trouve qu’une partie de ce surplus de population peut sans doute se consacrer à l’économie souterraine (la France s’apprête à intégrer dans le calcul de son PIB les revenus du trafic de drogue ou de la prostitution225). Mais que faire du reste? Il y a d’abord la prison ; mais une prison de masse conçue non pour discipliner mais pour retirer du monde, pour reléguer, pour parquer. Et il y a aussi l’élimination occasionnelle par une police surarmée. On voit donc concrètement se mettre progressivement en place, avec l’État pénal, une tendance des sociétés biopolitiques qui avait été esquissée par Foucault : l’élimination de fait d’une partie “inutile” de la population (une partie qu’on va considérer comme “dégénérée”, anormale). Le traitement des enfants par l’État pénal américain illustre la même tendance. Dans les sociétés disciplinaires, l’école ou l’atelier sont là pour préparer les corps à la discipline du travail afin que les enfants deviennent de futurs producteurs efficaces. Dans les sociétés biopolitiques les enfants sont une richesse, une ressource à gérer. Dites ainsi, les choses sont un peu caricaturales car il n’y a pas d’un côté des sociétés disciplinaires et d’un autre côté des sociétés biopolitiques, il n’y pas une époque disciplinaire qui précèderait une époque biopolitique. Les différents types de dispositifs s’emboîtent et se superposent. On peut cependant considérer que les mécanismes biopolitiques prennent du volume et de l’importance avec le libéralisme et le néolibéralisme (c’est du moins ce que suggère 224. Bertrand Ogilvie, L’
Homme je
table
. Essai sur
l’
exterminisme et la violence extrême, Éditions Amsterdam, Paris, 2012, p. 72
. 225.
Voir Romaric Godin « Intégrer
le trafic de drogue dans le PIB
, ou la
preuve des limites d’un fétiche é
conomique », article publié par Mediapart le 6 février 2018. 158
Foucault). Or, aux États-Unis, l’école devient un moyen d’alimenter la machine pénale226. En effet, certains États ont légiféré pour que l’absentéisme soit un délit et que les enfants puissent être poursuivis. Ainsi, par exemple, le Texas poursuit en correctionnelle les enfants de 12 à 18 ans qui manquent les cours. Et le nombre de condamnations de ces enfants (ils sont considérés comme majeurs, donc leurs parents ne peuvent intervenir ou les représenter) n’est pas marginal : « En 2013, le Texas a condamné 115 782 enfants pour “non-assiduité scolaire”, ce qui lui a permis d’engranger 16 millions de dollars de frais de justice et d’amendes. Le pourcentage de noirs et d’Hispaniques chez les enfants condamnés s’élève au chiffre remarquable, mais guère surprenant, de 83 %. »227 On le voit, des populations d’enfants deviennent un gisement, une ressource pour alimenter le financement de l’État pénal. Et, simultanément, certains d’entre eux sont très tôt identifiés comme faisant partie des individus potentiellement considérés comme inutiles et improductifs. Et d’emblée ces individus sont spécifiés selon des critères raciaux. En effet, pour les enfants comme pour l’ensemble des condamnés (ou pour les victimes de violences policières), l’État pénal américain présente un caractère racialement discriminant qui est fort impressionnant. La couleur de l’État pénal L’incarcération de masse qui, aux États-Unis, s’est mise en place à compter de la moitié des années 1970 possède une dimension fondamentale : elle frappe massivement la population noire. Au début des années 2000, on estimait qu’un jeune afro-américain sur trois était destiné à faire de la prison et que, dans les grandes villes, « plus de la moitié des jeunes adultes noirs se trouveront sous main de la justice – en prison, en conditionnelle ou en mise à l’épreuve »228. Tous les chiffres en témoignent : la justice, aux USA, a une couleur. Et ce sont les afro-américains qui sont criminalisés dans des proportions qui vont bien au-delà de ce que, proportionnellement, ils représentent dans la population américaine. Pour rendre compte de ce phénomène, on doit d’abord écarter deux mauvaises explications. 226. Depuis une loi de 2002, les lycées des quartiers les plus défavorisés proposent cependant une alternative à l’engrenage pénal : une option “armée” permet aux adolescents hispaniques ou noirs de se militariser et de s’engager très précocement dans les forces armées des États-Unis (voir ce documentaire : https://info.arte.tv/fr/usa-les-petits-soldats-de-lamerique). Pour les adolescents les plus défavorisés, aux États-Unis, le choix se réduit à l’alternative suivante : prison ou armée? 227. Keeanga-Yamahtta Taylor, Black Lives Matter, op. cit. p. 220. 228. Michelle Alexander, Incarcération de
masse
..., op. cit., p.
20.
159 La première mauvaise explication c’est celle qui part de l’idée, exacte et pertinente, que la criminalisation vertigineuse qu’ont connue les USA depuis les années 1970 vise les populations pauvres. Or, noirs et hispaniques, parce qu’ils sont surreprésentés parmi ces populations, seraient mécaniquement aussi surreprésentés parmi les condamnés du système pénal américain. Le problème, c’est que l’analyse des courbes de l’incarcération aux USA durant la période concernée ne cadre pas avec cette explication : on voit très vite progresser les incarcérations de jeunes noirs dans toute la période qui va jusqu’aux années 2000, avant une relative accalmie et une subite augmentation de l’incarcération des hispaniques. Il semble donc bien qu’il faille totalement inverser le raisonnement : l’État pénal américain est un dispositif pour maintenir la population noire dans la pauvreté ; et même pour aggraver cette pauvreté. La seconde mauvaise explication est plus perverse parce qu’elle donne un fondement à une espèce de sens commun raciste. Elle part de l’image d’Épinal que la société américaine s’auto-attribue : aux États-Unis, l’ascension sociale est toujours possible pour celle ou celui qui travaille courageusement. Donc, la réussite ou l’échec social de chacun est l’indice de son caractère. Il suffit de déplacer ce préjugé de l’individu au groupe social pour aboutir à une pure conception raciste : « par extension, l’échec d’un groupe racial à s’élever façonne très négativement l’image du groupe entier. »229 On l’a compris, si la population noire est surreprésentée parmi les individus incarcérés ou sous contrôle judiciaire, c’est que le “caractère” des afro-américains ne les porte pas à “s’en sortir” et qu’il condamne un grand nombre d’entre eux à rester pauvres. On reconnaîtra sans doute sous les sophismes qui conduisent à présenter un préjugé – historiquement formé et transmis – comme le constat d’une “nature”, une variation sur le thème de l’individu entrepreneur de lui-même dont Foucault nous dit qu’il est le principe organisateur de la pensée néolibérale : celui qui échoue n’est pas suffisamment entreprenant et responsable de lui-même. De là à penser que ce qui vaut pour l’individu mérite d’être étendu au groupe racial, il n’y a qu’un pas. Et ce pas peut être vite franchi. L’ethnicisation de la criminalité obéit donc à la logique inverse de la logique déterministe vu précédemment : il ne s’agit plus de dire que les noirs sont plus condamnés que les autres parce qu’ils sont plus pauvres, mais de dire qu’ils le sont parce ’ils ne font pas le nécessaire pour réussir dans une société qui serait angéliquement ouverte à l’ascension de tous ceux qui veulent réussir. 229. Ibid., p. 25. 160
Une fois ces deux raisonnements caricaturaux écartés, on est confronté à une explication simpliste et superficiellement arithmétique en vertu de laquelle il y aurait une augmentation constante de la criminalité qui aboutit mécaniquement à une inflation des incarcérations et à la nécessité d’inventer des dispositifs de contrôle sans cesse plus répressifs : « Comment s’explique cette hyperinflation carcérale? La première réponse, celle de l’idéologie dominante et de la recherche officielle, est de dire qu’elle est liée au crime. Mais la courbe de la criminalité a stagné de 1973 à 1993 avant de chuter fortement, au moment même où l’emprisonnement s’envolait. »230 Là encore on le voit, l’explication à prétention scientifique ne tient pas la route : il n’existe aucun lien entre le taux de criminalité et le taux d’incarcération. Ce qui, d’ailleurs, peut aussi ouvrir la porte à un discours parfaitement démagogique : si la criminalité régresse – ce qui est le cas aux USA depuis 1991 – c’est justement grâce à la politique pénale répressive qui a été mise en place. Le problème, c’est que l’on peut constater que les infléchissements respectifs de la courbe de l’incarcération et de celle de la criminalité (à la hausse pour l’une, à la baisse pour l’autre) ne se produisent pas, dans le temps, sur la même période. Il semble assez évident qu’il y a déconnexion entre ces deux courbes. Alors, que nous reste-t-il comme explication plausible? Nous devons nous ranger à l’idée que l’État pénal américain est un dispositif destiné à perpétuer la ségrégation raciale en la faisant passer par d’autres chemins que ceux qu’elle empruntait tant que l’égalité des droits n’était pas formellement acquise : « alors que la proportion des Noirs dans chaque “cohorte” de criminels est allée en diminuant pendant vingt ans, leur part de la population carcérale, elle, n’a cessé d’augmenter »231. En fait, il s’agit de continuer à faire exister et fonctionner la ségrégation sous le régime de l’égalité des droits : « La dure réalité est que, pour des raisons presque sans rapport avec les véritables caractéristiques de la criminalité, le système judiciaire américain est devenu un système de contrôle social unique dans l’histoire mondiale. L’ampleur de ce système pourrait faire croire qu’il touche la plupart des Américains, mais ses cibles principales sont essentiellement définies sur une base raciale. »232 Michelle Alexander explique que, paradoxalement, c’est l’indifférence à la couleur de peau qui permet à la ségrégation de se perpétuer : comment imaginer que l’État pénal est un système de ségrégation raciale puisqu’il vise à poursuivre et neutraliser la criminalité dans 230. Loïc Wacquant, « Le corps, le ghetto et l’État pénal. Entretien réalisé par Susana Durao », op. cit.. 231. Ibid. 232. Michelle Alexander, Incarcération de masse..., op. cit., p. 19. 161
une nation qui ne prête plus d’attention à l’ethnie et à la couleur de peau? D’un dispositif explicitement fondé sur la prise en compte de la couleur de peau, la ségrégation est passée à un dispositif qui exclut massivement toujours les mêmes (les afro-américains – auxquels, cependant, viennent progressivement s’agréger les hispaniques) mais sans avoir besoin de se focaliser explicitement sur la couleur de peau. On pourrait presque dire qu’on passe d’un dispositif brutalement disciplinaire à un dispositif plus “biopolitique” de gestion des populations. Et cela s’accompagne, sans surprise, d’un changement d’objectif du dispositif : le premier ambitionne de mettre à disposition de la production une main d’œuvre docile et peu coûteuse. Le second vise à isoler et marginaliser une population devenue inutile dont la production n’a plus besoin. C’est ce que formule très clairement Michelle Alexander pour résumer le chapitre 5 de son livre : « Le chapitre 5 explore également certaines différences entre l’esclavage, Jim Crow233 et l’incarcération de masse : tandis que cette dernière est conçue pour entasser une population considérée comme superflue, non nécessaire au fonctionnement de la nouvelle économie globale, les systèmes de contrôle antérieurs étaient destinés à exploiter et contrôler la force de travail noire. »234 Cela suggère au moins deux pistes de réflexion et d’analyse : la première et la plus vaste, c’est celle qui porte sur l’émergence contemporaine, dans les sociétés développées et “démocratiques” de cette idée qu’il existe une partie de la population qui est devenue « superflue ». Que faire de ce superflu? C’est manifestement une question souterraine du néolibéralisme. Et c’est probablement une des caractéristiques des formations sociales contemporaines dont Foucault avait eu l’intuition lorsqu’il affirmait brutalement que la gouvernementalité biopolitique s’accompagnait nécessairement du racisme235. La seconde piste porte sur la mise au jour des mécanismes de ce contrôle social qui relègue des populations au rang d’une humanité désormais en trop parce que sa force de travail est devenue inutile. C’est ce que disait Loïc Wacquant en 2008 : « il faut sortir du schéma “crime et châtiment” et repenser la prison comme une institution politique, une composante centrale de l’État. Et l’on découvre alors que le surgissement de l’État pénal est le résultat d’une politique de pénalisation de la misère qui répond à la montée de l’insécurité salariale et de l’effondrement du 233. Il s’agit du système de ségrégation qui a sévi dans les États du Sud des États-Unis jusqu’aux année
s 1960. 234. Michelle Alexander, Incarcération de masse..., op. cit., p. 30. 235. IFDS. À ce sujet, voir toute la fin du cours du 17 mars 1976, pp. 226-234. Nous y reviendrons dans la partie IV. 162 ghetto comme mécanisme de contrôle d’une population doublement marginalisée – au double plan matériel et symbolique.236 L’interprétation du sociologue vise donc à mettre en regard la montée de « l’insécurité salariale » avec celle de l’incarcération. On part donc bien de l’observation selon laquelle le néolibéralisme modifie le salariat parce qu’il constate que la force de travail d’une partie de la population n’est désormais plus suffisamment nécessaire pour qu’on la discipline ou qu’on la protège. Il convient alors de la gérer autrement, c’est-à-dire de la marginaliser et de la parquer dans des espaces de relégation. Bien entendu, la prison est un des premiers outils dont l’État peut alors user pour “traiter” cette population superflue. On a donc un mécanisme qui, simultanément, précarise le travail, crée l’insécurité sociale, génère la misère et invente l’incarcération de masse. Nous sommes donc très clairement confrontés à une régression de l’État social corrélative à un développement de l’État pénal. Dans une telle logique, on a à la fois une marginalisation matérielle (l’enfermement dans la prison, dans le ghetto, dans la misère) et une marginalisation symbolique (le racisme). Mais quels sont les mécanismes qui font fonctionner l’État pénal américain, ce « système de contrôle social unique dans l’histoire »? Pour les déterminer, il faut commencer par bien reconnaître que la machine à punir dont il est question ne se réduit pas aux seuls murs de la prison. La population qui est visée par la police, la justice et la prison est mise sous contrôle à l’intérieur comme à l’extérieur des lieux d’enfermement : « une fois libérés, les exdétenus pénètrent dans un monde occulte de discrimination légale et d’exclusion sociale permanente »237. C’est ce qui fait dire à Michelle Alexander que « le système d’incarcération de masse est fondé sur la marque de la prison, pas sur le temps passé en prison »238. L’État pénal américain a plusieurs facettes : D’abord l’incarcération en prison, bien entendu. Mais aussi, ensuite toute une gamme de discriminations : à l’emploi, aux contrats de location, aux aides au logement, aux demandes de prêts, aux inscriptions scolaires, aux allocations diverses. En effet, sur tous les formulaires, il y a toujours une « case à cocher » qui signale que vous avez déjà été condamné. Le lynchage n’existe plus mais cela ne met pas à l’abri de violences parfois extrêmes puisque la police doit contrôler et pourchasser tous ceux qui “ont l’air” de criminels. Vous êtes aussi prisonnier de la dette carcérale (que nous avons évoquée précédemment).
236. Loïc Wacquant, « Le corps, le ghetto et l’État pénal. Entretien réalisé par Susana Durao », op. cit.. 237. Ibid. p. 24. 238. Ibid., p. 26. 163
On vous retire le droit de vote (durant l’incarcération, mais aussi après, voire à vie dans quelques États). Si vous ajoutez à cet arsenal répressif la réforme Clinton de la protection sociale de 1996, vous entrez dans une boucle infernale : il faut travailler pour bénéficier d’allocations. Or, votre condamnation vous exclut presque à coup sûr du marché du travail et vous cantonne à l’économie souterraine. De plus, la loi TANF (Temporary Assistance for Needy Family Program) interdit toute aide sociale sur fonds fédéraux aux condamnés pour faits de drogue239. Or, la majorité des condamnations sont des condamnations pour faits de drogue, surtout dans la population noire ou hispanique. Il apparaît donc que le “modèle américain” dont parlaient Bourdieu et Loïc Wacquant dans Le Monde diplomatique s’organise autour du thème de l’insécurité. Mais l’insécurité que l’incarcération de masse prétend combattre n’est que le rideau de fumée qui cache l’insécurité sociale. Et cette insécurité sociale n’est pas la conséquence des caprices de quelques “souverains” dissimulés dans des lieux de pouvoir. C’est le corrélat nécessaire du fait qu’une partie – de plus en plus significative – de la population mondiale devient superflue parce que sa force de travail ne peut plus trouver à se vendre dans l’économie globalisée contemporaine. Le capitalisme peut se passer de plus en plus de bras. Et il ne s’agit pas seulement de viser ceux qui résident en des territoires éloignés et qui vivent hors de cette économie globalisée – bien que, quand ceux-là quittent ces territoires, il convienne de les stopper, de les ignorer, de les enfermer, de les laisser se noyer, de les traiter comme une menace pour “la” sécurité. Il s’agit aussi de gérer ceux, de plus en plus nombreux, qui deviennent inutiles à l’intérieur même des pays développés où règnent la marchandise et son fétichisme. À ces derniers on peut même commencer à appliquer une logique d’incarcération en criminalisant les mouvements sociaux – des mouvements sociaux qui sont eux-mêmes mécaniquement alimentés par cette insécurité sociale “de l’intérieur”.
L’insécurité salariale
Cette explosion de l’incarcération aux États-Unis durant les décennies 1980 et 1990 s’est accompagnée d’une pression très forte sur le monde du travail. On peut même affirmer que le mécanisme général de toute cette évolution peut se résumer ainsi : le capitalisme peut – et même doit – se passer de plus en plus de bras. Et cela a deux conséquences : 239. Ibid. Sur tous ces aspects « techniques » des mécanismes de l’État pénal américain, voir pp. 201-228. 164 1/ Les employeurs vont pouvoir exercer une forte pression sur le travail : réduction des salaires, réduction du droit du travail, réduction du (peu de) pouvoir des syndicats. 2/ Pour la frange de la population appauvrie qui est encore en surplus au regard de cette compression du travail, il va y avoir l’enfermement. Par conséquent, il s’agit bien d’un double phénomène : appauvrissement ’un côté et enfermement punitif de la pauvreté de l’autre. Le néolibéralisme fabrique la misère qu’il punit. Il faut commencer par le début, c’est-à-dire le sort fait aux travailleurs dans le “modèle” américain. L’usage du mot “modèle” est ici à prendre au pied de la lettre. En effet, à la fin des années 1990, le prêchi-prêcha néolibéral ne cessait de vanter le modèle américain, notamment en faisant valoir un taux de chômage environ trois fois plus faible que celui de la France, par exemple. C’est donc toute l’Europe qui était invitée à prendre le chemin ouvert par les États-Unis ; et c’est effectivement ce chemin qu’elle a emprunté. Tout s’est donc passé comme si l’on pouvait considérer la société américaine, à la fin du XXe siècle, comme une sorte de « laboratoire social »240 du néolibéralisme (et pas seulement de laboratoire pénal dans le traitement de la misère). Qu’en est-il du taux de chômage aux États-Unis à la fin du XXe siècle (un taux tant et tant utilisé pour stigmatiser la fabrique à chômeurs que serait la protection sociale en Europe)? La comparaison des taux des différents pays européens avec celui des USA ne permet pas de dégager une conclusion claire puisque l’Autriche, la Suisse, les Pays-Bas ou la Norvège présente un taux inférieur à celui des États-Unis. Et si l’on pousse un peu l’analyse, on constate que le taux affiché par les USA est obtenu grâce à des modifications décisives de la structure du monde du travail : en 1999, huit millions d’américains occupent deux emplois ou plus. Cela signifie que les emplois précaires se sont multipliés. De plus, de nombreux emplois à temps partiel sont dissimulés : Wal-Mart, Starbucks ou Mc-Donald estiment qu’un emploi à temps plein est de 20 heures ou 28 heures. Cela permettait d’afficher des statistiques reluisantes qui faisaient pâlir d’envie les États qui n’ont pas encore suffisamment détruit leurs propres systèmes de protection sociale. Mais la conséquence de cette précarisation, c’est que, en 1999, « un travailleur sur quatre exerce un emploi atypique »241. On peut aussi s’apercevoir que le taux de chômage est positivement affecté par... l’emprisonnement massif d’une partie 240.
Rick Fantasia, « Dictature sur le prolétariat. Stratégies de répression et travail aux États-Unis », Actes de la recherche en sciences sociales, 2001/3, n° 138, p. 3. 241. Ibid., p. 4. Pour toutes les données chiffrées, l’article de Rick Fantasia dresse une synthèse saisissante, et fort utile, de l’état du monde du travail après deux décennies d’expériment
ation sociale néolibérale. 165 de la jeunesse (noire et hispanique) : ces jeunes disparaissent des statistiques pendant qu’ils sont en cellule. Mais qu’en est-il des salaires? La réponse est simple : même si la situation a été légèrement corrigée après 1995, et parce que « le revenu hebdomadaire moyen de 80 % des Américains, jusque-là ajusté à l’inflation, a chuté de 18 % entre 1973 et 1995 », il faut admettre que les travailleurs ont vu leur sort se dégrader significativement. Dans le détail, seuls les cadres dirigeants, les professions libérales et les techniciens ont vu leur revenu augmenter. Plus spectaculaire encore : « Pendant cette période la proportion des travailleurs dont le salaire se situait au seuil de pauvreté s’est élevée dans tous les secteurs et a été particulièrement importante chez les Hispaniques et les Noirs. Entre 1973 et 1999 le pourcentage d’Hispaniques de sexe masculin gagnant un salaire égal au seuil de pauvreté s’éleva de 25,1 % à 40,3 % ; le pourcentage de Noirs de 24,8 % à 29,5 % ; et le pourcentage de Blancs de 10,7 % à 16,1 %. »242 On constate aussi que moins le niveau de formation est élevé, plus la baisse du revenu salarial est importante. On est donc très clairement dans une logique inégalitaire qui appauvrit les plus pauvres. On voit aussi que l’assurance maladie patronale (si importante dans un pays où le système de santé privé coûte très cher) est en régression forte. Idem pour le financement des retraites par les employeurs. Sur la même période, les États-Unis sont le seul pays de l’OCDE à avoir vu leur temps de travail annuel augmenter. Le tableau est un peu fastidieux à dresser, mais tout converge dans la même direction : appauvrissement et précarisation du salariat. Des réalités que le “modèle” va transmettre aux pays européens. Il n’est par conséquent pas si difficile que cela de comprendre la dynamique néolibérale : dans le cadre d’un monde “ouvert”, la concurrence a d’abord la vertu d’exercer une pression à la baisse sur le coût du travail. Dans les pays développés, elle permet alors de pouvoir se passer de la force de travail de certains et de rémunérer celle des autres à bas prix ; elle autorise aussi la généralisation d’une précarité de plus en plus asphyxiante. Que faire alors de la frange misérable de population dont ce processus rend l’existence superflue? Que faire de ceux dont la force de travail est peu ou pas du tout “employable”? Réponse : la criminaliser et l’emprisonner. Tout se passe donc comme si l’on assistait dans le capitalisme à une sorte d’importation des “exclus” : pendant longtemps ils existaient uniquement en périphérie, loin des pays développés, c’est-à-dire loin du centre. Désormais, ils sont aussi au 242. Ibid., p. 5. 166 cœur des métropoles impérialistes. On est toujours dans un impérialisme d’exclusion243, mais les exclus sont désormais aussi au cœur des pays développés, ils sont autant au centre qu’en périphérie. Ce sont les exclus de l’intérieur. Quant aux exclus de la périphérie, ils ne sont, bien évidemment, absolument pas les bienvenus dans les pays du centre : ces derniers ont “leur” misère à gérer, c’est-à-dire leurs humains superflus à traiter. Ils ne peuvent accepter d’accueillir toute la misère du monde. La “leur” leur suffit amplement pour peupler leurs ghettos, leurs quartiers de relégation ou leurs prisons. C’est la forme contemporaine que le néolibéralisme donne à ce que Kurz décrit comme la caractéristique du capitalisme, à savoir « la constitution d’un espace d’inclusion excluante et d’exclusion incluante qui [...] se présente comme l’espace de la consomption, aux fins de l’économie d’entreprise, d’énergie abstraitement humaine »244.
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Editorial: Alternative Regenerative Medicine for Diabetes: Beyond the Stem Cell Approach
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Abbreviations: T1DM, type 1 diabetes mellitus; T2D, type 2 diabetes; AHST, autologous non-myeloablative hematopoietic
stem-cell transplantation; IDAA1c, insulin dose adjusted A1c; MC, microvascular complications; ECM, extracellular matrix;
iPS, Induced pluripotent stem cells; OB/RM, organ bioengineering and regenerative medicine. INTRODUCTION Despite increasing awareness of the problem, diabetes still presents a challenge not only for patients
but also for clinicians and other professionals who have to diagnose, monitor, and treat it knowing
its many facets. In the last few decades, diabetes has become rampant in the world with unstoppable
growth everywhere but especially in developed countries, coupled and supported by an escalating
global epidemic of overweight and obesity - (globesity) which is taking over many parts of the world
(1). Insulin daily use is the most widespread treatment for type 1 diabetes mellitus (T1DM) while
oral hypoglycemics are the first choice in the treatment of type 2 diabetes (T2D). Although these
measures have revolutionized diabetes management, they remain sub-optimal treatments. To date,
beta cell replacement through pancreas or islets transplantation is considered the only definitive
treatment bringing diabetic patients to independence from exogenous insulin administration and
increasing both survival and quality of life. However, this solution has several limitations notably the
shortage of organs or the need for chronic immunosuppressive therapies that are not without risk. EDITORIAL
published: 09 July 2021
doi: 10.3389/fendo.2021.648763 published: 09 July 2021
doi: 10.3389/fendo.2021.648763 Editorial on the Research Topic Alternative Regenerative Medicine for Diabetes: Beyond the Stem Cell Approach Edited and reviewed by:
Hans Ulrich Häring,
Tübingen University Hospital,
Germany
*Correspondence:
Andrea Peloso
Andrea.Peloso@hcuge.ch Edited and reviewed by:
Hans Ulrich Häring,
Tübingen University Hospital,
Germany In this context, dynamic developments in the field of regenerative medicine open up new
horizons and bring new hopes in the treatment of diabetes. *Correspondence:
Andrea Peloso
Andrea.Peloso@hcuge.ch This Research Topic draws together two original articles (Penaforte-Saboia, Montenegro et al.;
Penaforte-Saboia, Couri et al.), one review article (Peloso et al.) and one commentary article
(Cobianchi et al.) in order to explore potential alternative strategies for T1DM. Specialty section:
This article was submitted to
Clinical Diabetes,
a section of the journal
Frontiers in Endocrinology
Received: 01 January 2021
Accepted: 21 May 2021
Published: 09 July 2021 Specialty section:
This article was submitted to
Clinical Diabetes,
a section of the journal
Frontiers in Endocrinology As one of the leading groups in autologous non-myeloablative hematopoietic stem-cell
transplantation (AHST) in T1DM, Penaforte-Saboia et al. assessed the long-term frequency of
microvascular complications, beta cell function, and glycemic control in a group of 24 T1DM
patients after AHST. Results were associated with data obtained from 144 aged and gender-matched
diabetic patients, treated with conventional therapy (CT). In their analysis, the occurrence of
microvascular manifestations in transplanted versus CT patients, was lower while a higher residual
beta cell function and better glycemic control compared to the CT group was detected. Received: 01 January 2021
Accepted: 21 May 2021
Published: 09 July 2021 Editorial: Alternative Regenerative
Medicine for Diabetes: Beyond the
Stem Cell Approach Department of Visceral and Transplantation Surgery, University of Geneva Hospitals, Geneva, Switzerland Keywords: regenerative medicine, pancreas bioengineering, diabetes, type 1 diabetes mellitus, organ
bioengineering and regeneration ACKNOWLEDGMENTS I thank all the authors who have made a contribution to this
Research Topic with their articles. We would also like to thank all
the reviewers who kindly agreed to review the manuscripts
submitted to this Research Topic and provided important
remarks to enhance the publications’ quality. Conflict of Interest: The author declares that the research was conducted in the
absence of any commercial or financial relationships that could be construed as a
potential conflict of interest. Citation: Peloso A (2021) Editorial: Alternative
Regenerative Medicine for Diabetes:
Beyond the Stem Cell Approach. Front. Endocrinol. 12:648763. doi: 10.3389/fendo.2021.648763 July 2021 | Volume 12 | Article 648763 Frontiers in Endocrinology | www.frontiersin.org 1 Editorial: Regenerative Medecine for Diabetes Therapy Peloso Peloso The same group investigated, in a cross-sectional study, the
correlations between insulin dose-adjusted A1c (IDAA1c)
(intended as an easy and fast alternative to evaluate pancreatic
b-cell function) and microvascular complications (MC),
including diabetic retinopathy, neuropathy, and nephropathy
(Penaforte-Saboia et al.). The authors report that, in a
representative Brazilian population of T1DM patients, those
with IDAA1c ≤9 presented a lower frequency of MC, as well
as fewer episodes of hypoglycemia, in the month prior to the
analysis. To our knowledge this is one of the first articles
correlating IDAA1c with diabetic microvascular complications
and hypoglycemic events in patients with T1DM, which includes
a large number of patients undergoing immunomodulatory
therapy with autologous non-myeloablative hematopoietic
stem-cell transplantation. In total, 144 diabetic patients
received conventional therapy (CT) and 24 patients were
treated with autologous nonmyeloablative hematopoietic stem-
cell transplantation (AHST). These two groups were further
subdivided according to their IDAA1c values (30 patients had
IDAA1c ≤9; 138 had IDAA1c >9). Then, the prevalence of MC
and hypoglycemia were compared between the groups. Finally in
this representative population of T1DM patients, patients with
IDAA1c ≤9 presented a lower frequency of MC, as well as fewer
episodes of hypoglycemia, in the month prior to the analysis. been reported as a scaffold solution for organ bioengineering and
regenerative medicine (OB/RM) approaches for the treatment of
juvenile (type 1) diabetes mellitus. In particular, ECM-derived
scaffolds have been extensively explored, showing their potential
to be merged to a beta cell source to address the shortage of
transplantable organs. This cell-on-scaffold technology stems
from the ability to remove cells from a tissue, leaving the ECM
almost intact in terms of tridimensional architecture and
biological cues. Finally the commentary offered by Cobianchi et al. covers one
of the most promising approaches for beta-cell replacement: the
3D cell aggregate system. This system has been used by Lebreton
et al. to manufacture insulin-producing engineered organoids
(2). The commentary highlights the importance of the use of
dissociated islands and human amniotic epithelial cells (hAECs);
a breakthrough in this field. AUTHOR CONTRIBUTIONS In our review (Peloso et al.), we provided an overview of the
existing knowledge of current experimental strategies in the
treatment of diabetes covered by the umbrella of regeneration. In particular, stem cell reprogramming, extracellular matrix
(ECM) scaffolds, and insulin-like organoid technology have the
potential to overcome the actual limitations related to pancreas
transplantation. Through an unceasing deep understanding of
beta cell and pancreatic development, stem cell research
continues to garner interest as a future cure of T1DM. Induced
pluripotent stem cells (iPS) are demonstrated to be a potential
unprecedented beta cell source via their capacity to differentiate
into all major somatic cell lineages (Peloso et al.). In the last
decade, several natural- and synthetic-derived polymers have AP edited and revised the manuscript. The author confirms
being the sole contributor of this work and has approved it
for publication. Citation: In conclusion, the articles in this Research Topic not only
provide important insights concerning T1DM (Penaforte-
Saboia, Montenegro et al.; Penaforte-Saboia, Couri et al.), but
also explore new strategies for beta cell replacement (Peloso et al.;
Cobianchi et al.) (2). Conflict of Interest: The author declares that the research was conducted in the
absence of any commercial or financial relationships that could be construed as a
potential conflict of interest. July 2021 | Volume 12 | Article 648763 Frontiers in Endocrinology | www.frontiersin.org 2. Lebreton F, Lavallard V, Bellofatto K, Bonnet R, Wassmer CH, Perez L, et al.
Insulin-Producing Organoids Engineered From Islet and Amniotic Epithelial
Cells to Treat Diabetes. Nat Commun (2019) 10(1):4491. doi: 10.1038/s41467-
019-12472-3 REFERENCES 1. World Health Organization. Global Strategy on Diet, Physical Activity and Health. WHO Library Cataloguing-in-Publication Data World Health Organization. Global strategy on diet, physical activity and health. 1. Diet 2. Exercise 3. Health
promotion 4. National health programs 5. International cooperation I. Available
at: www.who.int/activities/controlling-the-global-obesity-epidemic. Copyright © 2021 Peloso. This is an open-access article distributed under the terms of
the Creative Commons Attribution License (CC BY). The use, distribution or
reproduction in other forums is permitted, provided the original author(s) and the
copyright owner(s) are credited and that the original publication in this journal is
cited, in accordance with accepted academic practice. No use, distribution or
reproduction is permitted which does not comply with these terms. 2. Lebreton F, Lavallard V, Bellofatto K, Bonnet R, Wassmer CH, Perez L, et al. Insulin-Producing Organoids Engineered From Islet and Amniotic Epithelial
Cells to Treat Diabetes. Nat Commun (2019) 10(1):4491. doi: 10.1038/s41467-
019-12472-3 July 2021 | Volume 12 | Article 648763 Frontiers in Endocrinology | www.frontiersin.org
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Insights into the embryo/endosperm interface in Zea Mays and Arabidopsis thaliana. Life Sciences [q-bio]. ENS de Lyon, 2019. English. ⟨NNT : 2019LYSEN015⟩. ⟨tel-04435039⟩
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(Kawashima and Goldberg, 2010; Randolph, 1936). In Arabidopsis, the suspensor also plays critical roles in transporting auxin to and from the apical part of the embryo, and thus in embryonic patterning (Friml et al., 2003; Robert et al., 2013) However, in maize, the function of the suspensor remains unclear. The apical cell divides to give rise to the embryo proper. In the embryo proper the cells in contact with the surface of the embryo, called the protoderm give rise to the epidermis (Figure 2C). Subsequently, the inner tissues give rise to the precursors of vasculature and ground tissues. The three main plant cell types are thus determined in the early embryo proper (Esau, 1977; Hove et al., 2015; Jürgens, 1992). In Arabidopsis, the upper cells of the ground tissue expressed the stem cell marker gene and will give rise to the shoot apical meristem (SAM), while the root apical meristem is determined from the upper cell of the suspensor (RAM) (Hove et al., 2015). In both species, the overall radial symmetry of the embryo proper shifts to a bilateral symmetry during the transition stage (Jürgens, 1992; Jürgens et al., 1991; Van Lammeren, 1987). From this stage onwards the embryogenesis of Arabidopsis and maize, appears to diverge. In Arabidopsis two cotyledons emerge at the apical pole of the embryo from each side of the SAM (Figure 2D) (Hove et al., 2015; Jürgens, 1992; Lau et al., 2012). In maize, the embryo proper takes on a shield-like shaped. The precursors of the SAM are specified on the adaxial face of this structure, while the RAM precursors arise within the embryo (Zimmermann and Werr, 2005, 2007). Both the RAM and the SAM are then surrounded by protective organs, the coleorhiza and the coleoptile respectively. The abaxial face of the embryo differentiates into the scutellum, often considered as the single cotyledon of maize. Unlike the situation in Arabidopsis where the SAM and RAM remain largely inactive before the germination, the maize SAM generates several embryonic leaves prior to seed maturity (Figure 2B) (Randolph, 1936). The morphogenetic phase of embryogenesis require a huge network of genes involved in the specification of each territory (Hove et al., 2015; Palovaara et al., 2017). This phase is followed by a phase of growth and nutrient accumulation. The embryo grows at the expense of the surrounding endosperm. In Arabidopsis, the embryo replaces almost all of the endosperm and occupies, at maturity, most of the volume of the seed. The mature embryo consists of two large cotyledons and a hypocotyl (Baud et al., 2002) (Figure 1A). In contrast the mature maize 29 embryo occupies only around a third of the kernel (Figure 1B). In both species the mature embryo dehydrates and enters into slowed down long term life at the end of seed development. Germination occurs only when favourable environmental conditions are encountered. However whereas during the germination of Arabidopsis all the embryonic tissues emerge from the seed, in maize, the scutellum remains inside the seed where it acts almost like a haustorium, absorbing nutrients from the hydrolysing endosperm. I-2-D) The embryo and endosperm engage in extensive crosstalk during development
Endosperm is often considered as a reserve tissue for the embryo. This is, of course, true, particularly in the case of maize during germination when a significant flux of nutrients flows from the endosperm to the scutellum furnishing the energy for the growth of the young seedling. The endosperm probably also provides huge quantities of nutrients to the embryo during seed development, although these processes are surprisingly under-studied. However the embryo and endosperm are also strongly interdependent for developmental processes. Many studies of mutants highlight this interdependence in both Arabidopsis and maize. One striking example is the study of genetic translocation, especially in maize. As the embryo and endosperm have the same genetic alleles, a mutation in one compartment is usually also present in the other. However, translocations can produce seeds in which one compartment is genetically different to the other. For many mutants affecting maize kernel development, a mutation in one of the two compartments lead to a non-autonomous effect on the other (Chang and Neuffer, 1994; Mimura et al., 2018; Neuffer and Sheridan, 1980). For example the SHOHAI1 (SHAI1) putative transcription factor, involved in both embryo and endosperm patterning, has a reduced mutant phenotype in the embryo when SHAI1 is not mutated in the endosperm (Mimura et al., 2018). Mutants of genes expressed in only one compartment can also give indications about the inter-dependence of the compartments. In Arabidopsis, an excellent example of the effects of endosperm defects on the embryo is shown in the zhoupi (zou) mutant. ZOU encodes a transcription factor expressed specifically in the endosperm, but mutants show strong phenotypes for both endosperm and embryo. The endosperm is persistent due to a pleiotropic syndrome that includes a lack of cell wall modification leading to a lack of endosperm degradation. Consequently, the embryo remains very small and appears morphologically delayed, although it apparently matures normally at the metabolic and physiological level, as it undergoes desiccation and retains viability at germination. This demonstrates the strong effect 30 of endosperm on embryo development (Fourquin et al., 2016; Kondou et al., 2008; Yang et al., 2008). Furthermore, the absence of the embryo affect significantly the dynamic and patterning of the endosperm, although it can start to develop quite normally (Ohad et al., 1996). For example in maize, embryoless mutants do not develop a normal ESR, and in Arabidopsis when embryos die at early stages, endosperm degradation does not occur (Fourquin et al., 2016; Ogawa et al., 2015; Opsahl erstad et al., 1997). These examples indicate that crosstalk between the embryo and the endosperm underlies the harmonious development of both tissues, and thus seed viability. Although relatively little is known about the molecular mechanisms underlying embryo-endosperm crosstalk, we can assume that much of it takes place at the interface between the two tissues (Widiez et al., 2017). Our current knowledge of this interface will be described in next section. II) THE ARABIDOPSIS AND MAIZE EMBRYO/ENDOSPERM INTERFACE II-1) Definition of embryo/endosperm interface II-1-A) Delimitation in space
According to Oxford English dictionary, an interface is “a point where two systems, subjects, organizations, etc. meet and interact’’. In both Arabidopsis and maize, there are no symplastic connections between embryo and endosperm and consequently, so the embryo and endosperm are symplastically separate compartments (Schel et al., 1984; Stadler et al., 2005). As a consequence, the embryo/endosperm interface is composed of an apoplast separating both compartments. We also consider also that the layer of cells on either side of this apoplast are components of the interface, as well as the molecules crossing the interface.
II-1-B) Delimitation in time
The embryo and endosperm are generated by the double fertilization event, and the interface between the egg cell and the central cell will consequently not be addressed. In mature seeds, at least one layer of endosperm cells is maintained around the embryo, meaning that an intact embryo/endosperm interface remains present until germination. The embryo/endosperm 31 interface should be lost at germination when the embryo germinates, at least in Arabidopsis. However the post germination fate of the interface elements will also be discussed. II-2) Structure of the interface II-2-A) Cells on both side II-2-A-a) The epidermis and the suspensor on the embryo side
During the first steps of embryogenesis all embryonic cells are in contact with the endosperm but cell divisions rapidly lead to the isolation of inner embryonic cells from the endosperm. The cells at the surface of the embryo proper, which remain in contact with the endosperm, form a specialized cell layer, the protoderm, which ultimately gives rise to the epidermis (Jürgens, 1992; Randolph, 1936). Protodermal identity may be the “basal” cell fate of the embryo-proper, since epidermal markers are expressed from the two cell stage onwards, at least in Arabidopsis, although this may not be the case in maize (Abe et al., 2003; Ingram et al., 2000; Lu et al., 1996). However, the protoderm is officially recognised as a cell layer at the 16 cell stage in Arabidopsis and in the 6 DAP embryo in maize (Hove et al., 2015; Javelle et al., 2011). From these stages onwards, protoderm cells divide predominantly anticlinally to give rise to other protoderm cells which form a single cell layer. One of the defining features of these cell is the formation of the embryonic cuticle, which will be described later (Delude et al., 2016). In Arabidopsis, two redundant transcription factors are considered as the master regulators of the epidermal cell fate, Arabidopsis thaliana MERISTEM LAYER1 (ATML1) and PROTODERMAL FACTOR2 (PDF2) (Abe et al., 2003). Both are expressed before protoderm formation sensu stricto but their expression becomes restricted to this layer when internal cells appear (Hove et al., 2015). The double mutant atml1 pdf2 is lethal because the embryo arrests at the globular stage, highlighting the key role of the epidermis in early development (Ogawa al., 2015; San-Bento et al., 2014). Other genes that are involved in embryo epidermis cell fate or maintenance include the receptor kinase encoding CRINKLY 4 (CR4) gene in maize, and its orthologue ACR4 in Arabidopsis, the PHYTOCALPAIN-encoding DEFECTIVE KERNEL1 (DEK1) gene in both species and the receptor kinase-encoding ABNORMAL LEAF SHAPE 2 (ALE2) gene in Arabidopsis (Becraft et al., 1996, 2002; Johnson et al., 2005; Tanaka et al., 2007; Watanabe et al., 2004). From early stages of embryo development to the germination, the epidermis constitutes a specialized single cell layer defining the interface of the embryo with 32 the surrounding endosperm. The suspensor forms a different kind of interface. At the difference of the epidermis, cells neither adopt a pavement morphology nor are surrounded by cuticle. These differences suggest an alternative functionality of this interface that will be discussed later (§ II-3). Interestingly, not all the suspensor is in contact with the endosperm, as the most basal cell is in contact with the maternal tissues (Bowman, 1994; Robert et al., 2018; Van Lammeren, 1987). In Arabidopsis, the suspensor cell specification is under the control mitogen-activated protein (MAP) kinase kinase kinase gene YODA (YDA) which controls the suspensor identity via the regulation of several kinases and the WUSCHEL-RELATED HOMEOBOX 8 and 9 (WOX8 and WOX9) transcription factors (Lukowitz et al., 2004; Musielak and Bayer, 2014). YDA is activated by the membrane associated kinase SHORT SUSPENSOR (SSP) and putatively by exogenous peptides that will be described later (§ II-3-A) (Bayer et al., 2017; Costa et al., 2014). Suspensor patterning is also dependant of the plant hormone auxin (Bayer et al., 2017; Friml et al., 2003; Robert et al., 2018) Auxin comes first from the maternal tissues, before being produced by the scutellum and transported by PIN7 towards the embryo proper, where it influences cell patterning. At the early globular stage, a switch in the auxin flux appears at the is transported to the suspensor, where it is proposed to regulate suspensor development (Robert et al., 2013, 2018). The suspensor is thought to be mainly involved in early seed development because it undergoes cell death at later stages. In maize, suspensor degenerates from 14 DAP and in Arabidopsis at the bent-cotyledon stage (Blanvillain et al., 2011; Giuliani et al., 2002). II-2-A-b) The endosperm cells at the embryo interface
In contrast with the situation in the embryo, the endosperm cells at the interface show extremely dynamic behaviour during seed development. The endosperm surrounding the embryo consists first of the coenocyte. In maize, following cellularization, the endosperm cells near to the embryo differentiate into ESR cells. As mentioned above (§ I-1-C-b), ESR cells are small with a big nucleus and a dense cytoplasm rich in endoplasmic reticulum (OpsahlFerstad et al., 1997). The ESR is a transient tissue, from which the embryo emerges by elongation and which, as a result, becomes restricted to the basal pole of the embryo before degenerating and disappearing at around 15 DAP (Figure 2A,B). Unlike for other endosperm tissues in maize the genetic control of ESR specification, differentiation is still unclear. However ESR degradation appears to be regulated by ZmZOU, the single ortholog of ZOU in maize (Grimault et al., 2015). In 33 addition to their characteristic morphology, ESR cells are characterized by the expression of specific genes and thus appear to have their own transcriptional program (Li et al., 2014; OpsahlFerstad et al., 1997; Zhan et al., 2015). In Arabidopsis it is unclear whether a direct equivalent of ESR exists and the cell surrounding the embryo do not appear to have a particular morphology. Nonetheless the endosperm in contact with the embryo expresses a specific set of genes such as those encoding the ZOU transcription factor and the AtSUC5 sucrose transporter (Baud et al., 2005; Yang et al., 2008). Unlike the situation in maize, the transcriptionally defined zone surrounding the embryo moves as the embryo grows inside the endosperm and is therefore not restricted to a group cells. Endosperm cells in contact with the embryo degenerate progressively during seed development and are replaced by the growing embryo (Fourquin et al., 2016). In both Arabidopsis and maize the growth of the embryo leads to the appearance of new interfaces. In maize, after around 7 days, the embryo emerges from the ESR and makes contact with the starchy endosperm at the abaxial side and with the aleurone (a zone called the scutellar aleurone in barley), at the adaxial side (Figure 2B) (Jestin et al., 2008). Little is known about the endosperm cells that are adjacent to the scutellum, although cytological analyses have revealed some starchy endosperm characteristics (Van Lammeren, 1987). Cells were shown to collapse, leading to an accumulation of cell walls at the embryo interface, suggesting that cell death may occur and the embryo-endosperm interface penetrates into the endosperm as the embryo expands. However classical cytological studies (Evans blue staining studies) failed to reveal classical cell death characteristics, suggesting that cell death may be very localized to the interface with the embryo (Giuliani et al., 2002; Young and Gallie, 2000a, 2000b; Young et al., 1997). At the adaxial side, the scutellar aleurone consists of a single cell layer separating the embryo from the pericarp. The relationship between the scutellar aleurone and the embryo remains largely undiscovered. In the first chapter (section I-1) of this thesis the transcriptomic profile of these two interfaces will be characterized with a particular focus on the starchy endosperm cells adjacent to the scutellum. As in maize, the aleurone-like layer of the Arabidopsis endosperm makes the interface with the embryo, albeit relatively late in development. At maturity, the embryo is surrounded by the single layer of aleurone-like cell (Brown et al., 1999). II-2-B) Apoplastic components II-2-B-a) The cell wall
The cell wall between the egg cell and the central cell is formed during embryo sac cellularization (Russell, 1993). In maize, the egg cell shares a cell wall with central cell but also with the two synergids and is fused at one point with the embryo sac cell wall (Russell, 1993; Van Lammeren, 1987). Thus, after fertilization, the embryo is not entirely surrounded by endosperm and a connexion with the maternal tissues is maintained at the base of the suspensor. The cell wall between the egg cell and the central cell is discontinuous and large area of plasma membranes remain free between the two cells (Huang and Russell, 1992). Following the double fertilisation, cell wall material is massively produced by the egg cell. Evidence for this comes from in vitro studies of the fertilisation of maize protoplasts. Around 30s after fusion of the sperm cell with the egg cell, cell wall components are secreted, probably from vesicles stored under the plasma membrane. These components are sensitive to cellulases but not to pectinase, suggesting the presence of cellulosic material. The rapid cell wall production has been proposed to be a mechanism to avoid polyspermy (Kranz et al., 1995). As a result of this cell wall deposition, the cell wall of the young zygote becomes continuous, according to transmission electron microscopy (TEM) (Van Lammeren, 1987). The contribution of the endosperm post fertilisation is poorly documented, as is the composition of the embryo/endosperm cell wall. The embryo and endosperm are symplastically separated, indicating that the cell wall is devoid of plasmodesmata (Schel et al., 1984). As the embryo and endosperm stick together at young stages, the cell wall is considered as classical with two primary cell walls on either side stuck together by a middle lamellae, although little if any evidence exists to support this supposition. This putative middle lamella is supposed to be removed or remodelled during early development to be replaced by other apoplastic structures, although again no evidence for this has yet been reported. The situation in Arabidopsis is likely similar to that in maize, although very few ultrastructural studies of very early embryogenesis have been carried out in this species.
II-2-B-b) The embryonic cuticle
In the embryo/endosperm cell wall, TEM pictures reveal the appearance of an electron dense deposit during seed development. In Arabidopsis, this deposit appear at globular stage and forms a continuous layer by the early heart stage, while in maize a continuous layer is observed 9 DAP on the surface of the emerging scutellum (Creff et al., 2019; Schel et al., 1984). 35 This structure is called the embryonic cuticle. In Arabidopsis this cuticle is the precursor of that covering emerging organs post germination. In maize, little is known about embryonic cuticle establishment and its genetic control. A recent study showed that cuticle deposition on the coleoptile and first leaves is controlled by the MYC transcription factor FUSED LEAVES 1 (FDL1), but no study has been made of the scutellum cuticle (La Rocca et al., 2015). Embryonic cuticle has been studied in more detail in Arabidopsis. From the heart stage onwards, the whole embryo surface is surrounded by a continuous layer of cuticle, including the root tip, but with the exception of the suspensor (Berhin et al., 2019; Creff et al., 2019; Szczuka and Szczuka, 2003). The lipidic nature of the embryonic cuticle was showed by staining of the growing embryo surface with the lipophilic dye Auramine O (Rodkiewicz et al., 1994; Szczuka and Rodkiewicz, 1996; Szczuka and Szczuka, 2003). However it should be borne in mind that Auramine O also stains aromatic cell wall polymers (Considine and Knox, 1979; Nishikawa et al., 2005; Pesquet et al., 2005). Cuticle components are thought to secreted by the epidermis of the embryo, as highlighted by the expression of cutin biosynthesis genes in the embryonic epidermis from early stages onwards, and by the active secretion of cutinsomes that cross the cell wall and fuse with the cuticle (Creff et al., 2019; Stępiński et al., 2017). However, the fact that the embryonic cuticle is hidden inside the seed and is extremely thin makes investigation of its structure and composition extremely challenging. As a result cotyledon cuticles have been mainly used as a proxy. However this approach may be flawed as the cuticle is thought to be modified rapidly post germination, notably through the biosynthesis of cuticular waxes (Bassel et al., 2008; Todd et al., 1999; Winter et al., 2007). Genetic studies of mutants with defective cotyledon cuticles have led to the identification of several genes involved in embryonic cuticle establishment. Globally these genes encode proteins that fall into three classes: Enzymes required for cuticle biosynthesis, proteins (transcription factors and Receptor-Like Kinases) involved in specifying epidermal cell fate, and proteins involved in cuticle integrity control (Figure 3A). The first category is well known for post germination plant cuticles but remains poorly documented in the case of the embryonic cuticle (Delude et al., 2016). A recent study by the host team show that gpat4gpat8 double mutant, which lacks two glycerol-3-phosphate acetyltransferases, involved in the biosynthesis of monoacylglycerol cutin precursor, lacks various cutin components in newly emerged cotyledons and has a thinner and slightly defective (permeable) cotyledon cuticle (Creff et al., 2019; Jayawardhane et al., 2018; Li et al., 2007). Loss of function of proteins involved in epidermal specification/differentiation such as ATML1, PDF2, ALE2 or 36
A B Figure 3: (A) Scheme of the three genetic pathways controlling embryonic cuticle establishment. (B) Scheme of the signalling pathways controlling embryonic cuticle integrity and sheath production and deposition. 37 ACR4 also show defects in the cuticle but these are likely linked to a problems in cell identity, and highlight the very tight link between epidermal cell identity and cuticle biosynthesis (SanBento et al., 2014; Tanaka et al., 2007; Watanabe et al., 2004). Genes of the third category do not appear to directly control epidermal identity or cuticle biosynthesis and their function appears only to be needed for embryonic cuticle formation, but not for the formation of the cuticle of post germination organs (Creff et al., 2019; San-Bento et al., 2014). The first mutant of this class to be described was abnormal leaf shape 1 (ale1), which is defective for a gene encoding a predicted apoplastic protease of the subtilase family. Ale1 mutants have a discontinuous cuticle layer during the young stages of embryo development (Creff et al., 2019; Tanaka et al., 2001). Surprisingly ALE1 is expressed in the embryo surrounding region of the endosperm and appears to have a non-cell autonomous effect on the embryo (Tanaka et al., 2001; Yang et al., 2008). Another endosperm-specific gene was also found to be involved in this process: that encoding the ZOU transcription factor. As described before (§ I-1-D), zou mutants have a strong phenotype, and the most striking defect is the persistence of the endosperm and consequently, the production of a dwarf embryo. However, the embryo surface of zou mutants is also strongly affected, especially the embryonic cuticle (Kondou et al., 2008; Yang et al., 2008). Interestingly, ZOU controls ALE1 expression in the endosperm (Yang et al., 2008). Loss of function of another transcription factor, INDUCER OF CBF EXPRESSION 1 (ICE1) leads to a similar seed mutant phenotype to that seen in zou mutants. ICE1 make heterodimers with ZOU and is necessary for ZOU activity in the endosperm (Denay et al., 2014). However, as the embryonic cuticle is secreted by the epidermis, all these three endosperm genes must have a non-cell autonomous effect on the embryo. On the embryo side, loss of function of the two leucine rich repeat receptor like kinases (LRR-RKs), GASSHO1 and 2 (GSO 1 and 2), both expressed in the epidermis, leads to a double mutant phenotype with strong cuticle defects. GSO1 and GSO2 appear to act as two redundant paralogs since the two single mutants do not present any cuticle defects. (Creff et al., 2019; Tsuwamoto et al., 2008). The gso1 gso2 double mutant has a discontinuous cuticle but is not impaired in cutin biosynthesis or epidermal identity (Creff et al., 2019; San-Bento et al., 2014). The Mitogen-Activated Protein Kinase MPK6 has been proposed to act downstream of GSO1 and GSO2, and is phosphorylated in developing seeds in a partially GSO1/GSO2 dependent manner(Creff et al., 2019). TWISTED SEED1 (TWS1), an unknown protein whose loss of function phenotype strongly resembles that of gso1 gso2 mutants, and which is expressed in the embryo, has also been proposed to play a role downstream of GSO1 GSO2 signalling (Fiume 38 et al., 2016). GSO1 and GSO2 act in the same pathway as ZOU and ALE1 as demonstrated by epistasis analysis and the presence of overlapping transcriptomic changes in the respective mutants (Creff et al., 2019; Xing et al., 2013; Yang et al., 2008). The presence of a subtilase protease, a member of a protein family known to process apoplastic signalling peptides, in the endosperm, and of two LRR-RKs in the embryonic epidermis, suggests the presence of a molecular dialogue between the endosperm and the embryo. This dialogue presumably involves the production of a peptide, the identity of which was unknown at the start of my PhD. One plausible hypothesis is that this putative peptide is processed by ALE1 in the apoplast and then binds to the GSO receptors in the epidermis, triggering the closure of cuticle gaps (Figure 3B). Interestingly, two recent studies showed that GSO1, also known as SCHENGEN 3 (SGN3), as involved in Casparian strip formation in the root endodermis. In this context GSO1 binds the CASPARIAN STRIP INTEGRITY FACTOR 1 and 2 (CIF1 and 2) peptides which are produced in the stele (Figure 4). (Doblas et al., 2017; Nakayama et al., 2017). Gso1 (sgn3) and cif1 cif2 mutants have a patchy Casparian strip that is highly reminiscent of the patchy embryonic cuticle observed in the gso1 gso2 double mutant (Creff et al., 2019). Treatment of roots with active (processed) CIF1 and 2 peptides restores a continuous Casparian strip in cif1 cif2 but not in gso1 (sgn3) mutants, supporting the theory that perception of CIF1/2 peptides diffusing throughout the gaps of the Casparian strip by GSO1 mediates gap filling in the strip to form a continuous apoplastic structure (Doblas et al., 2017). Processed CIF peptides contain a sulphated tyrosine residue at the second position. Sulfation is catalysed by the TYROSYLPROTEIN SULFOTRANSFERASE (TPST), the unique tyrosine sulfotransferase encoded by the Arabidopsis genome (Komori et al., 2009). Tpst mutants also produce patchy Casparian strips, indicating that the sulfation is necessary for peptide efficiency (Doblas et al., 2017; Nakayama et al., 2017) (Figure 4). In addition to peptide sulfation, it is predicted that CIF1 and CIF2 propeptides are cleaved at the N-terminus of the active peptide (the active peptide being situated at the C-terminal extremity of the propeptide). However the peptidase involved has not yet been identified. At this point it is important to note that there are many similarities between the Casparian strip and embryonic cuticle: both are apoplastic structures produced do novo during plant development, and which must be intact before being functionally deployed (for example at germination for the embryo cuticle). The GSO1 receptor is involved in ensuring the integrity of both barriers, and loss of function of GSO1 (root) or GSO1 and GSO2 (embryo) gives mutant phenotypes that are similar, with gaps in the barrier. Importantly, in each case, pathway
39 stele endodermis cortex Figure 4: Casparian strip formation in the endodermis under the control of the SGSO1/SGN3 pathway. Upper panel represents the situation before the establishment of the apoplastic barrier and lower panel after. 40 components are expressed in spatially distinct patterns. So for example in the root the CIF1 and CIF2 peptides are produced specifically in the steel (on the inside of the barrier), whereas in the seed the subtilase ALE1 is produced only in the endosperm (on the outside of the barrier) (Doblas et al., 2017; Nakayama et al., 2017; Tanaka et al., 2001; Yang et al., 2008
). In both cases the G
SO
receptors are localised just adjacent to the
barrier
(Creff et al., 2019; Pfister et al., 2014; Tsuwamoto et al., 2008). Nonetheless, it appears that the peptides involved are not the same, since cif1 cif2 mutants have not been reported to have an embryonic cuticle phenotype (Nakayama et al., 2017). We postulate that GSO signalling pathway has been reused to ensure the integrity of two distinct apoplastic barriers but that the peptides involved differ. In chapter 3 of this thesis, the pathway controlling embryonic cuticle integrity will be investigated in Arabidopsis, with a focus first on TPST and then on the putative peptides binding the GSO receptors. II-2-B-c) Apoplastic deposits from the endosperm
In both Arabidopsis and maize, an intense secretion activity from the endosperm to the embryo/endosperm interface has been shown (Moussu et al., 2017; Van Lammeren, 1987). In maize, these secretions of unknown material were revealed by TEM analyses, 9 DAP (Van Lammeren, 1987). Later in development, a layer of fibrous material is deposited at the surface of the scutellum (Corona-Carrillo et al., 2014; Enriquez-Arredondo et al., 2005). This layer contains neutral lipids, phenolic molecules and callose like carbohydrates (Corona-Carrillo et al., 2014). However, the endosperm origin of this layer is not well established and the fibrous layer is probably a mix of embryo-derived deposits such as the embryonic cuticle and endosperm-derived deposits (Van Lammeren, 1987). Interestingly some crushed cell walls have been found in this layer, suggesting at least a partial contribution of the endosperm (Bourdu and Bousser, 1991). In Arabidopsis, a recent study by the host team characterized the endosperm deposits at the embryo/endosperm interface (Moussu et al., 2017). An amorphous material that is recognised strongly and specifically by two anti-EXTENSIN antibodies is produced in the endosperm cells surrounding the embryo and then deposited at the embryo/endosperm interface as the endosperm starts to break down (at around the early torpedo stage) (Smallwood et al., 1994, 1995). This material forms a continuous layer of around 200nm in thickness, on the outer (endosperm) surface of the embryonic cuticle, and this structure has been named the embryo sheath. Due to its strong labelling by anti-EXTENSIN antibodies, the embryo sheath is thought to be strongly enriched in EXTENSINS. Some genes were shown to be necessary for embryo 41 sheath formation and can classified in two categories. The first are the GSO1 GSO2 and ALE1 genes involved in the previously described cuticle integrity pathway. In gso1 gso2 and ale1 mutants, the sheath material is produced in endosperm cells but is not deposited at the embryo/endosperm interface and accumulates in the endosperm. Thus the sheath is not formed. The second class of genes appears to be involved in the production of sheath materials, and loss of function of these genes do not produce any materials in the endosperm that are recognised by anti-EXTENSIN antibodies. Two genes fall into this category: that encoding the transcription factor ZOU, which has many pleiotropic effects during seed development as previously described, and a gene encoding a cysteine rich peptide named KERBEROS (KRS). The expression of KRS in seeds is specific to the endosperm immediately surrounding the embryo, and is entirely dependent upon ZOU. Interestingly however, krs mutants do not show the pleiotropic phenotypes seen in zou mutants and KRS therefore seems to a specific regulator of sheath production. However the mechanism via which KRS production leads to sheath production and deposition is still unknown. Sheath composition and structure remains poorly understood for the moment. Moreover, the fate of the sheath post germination has not been investigated. In the second chapter of this thesis, I will specifically address these questions. II-3) What are the functions of the embryo/endosperm interface? II-3-A) Embryo/endosperm communication
The interdependent developmental programmes of the embryo and endosperm suggest a strong coordination of the development of these two compartments during seed growth. This implies the emission of a signals from one compartment, propagating to the adjacent compartment to trigger a developmental response. The symplastic separation of the embryo from the endosperm limits the potential communication mechanisms to those involving the apoplast (Schel et al., 1984; Stadler et al., 2005). Apoplastic communication can be split into two main types: mechanical and molecular. Mechanical communication is based on the detection of changes in mechanical forces (translated to strain), which can be caused by variations in turgor pressure or cell wall stiffness for example. Mechanical communication is thought to be particularly important for embryo/endosperm communication. Indeed, in Arabidopsis, when the embryo has a reduced growth, the expression of the classical cell death markers PLANT ASPARTIC PROTEASE A3 (PASPA3) and BIFUNCTIONAL NUCLEASE 1 (BFN1) in the endosperm is 42 strongly retarded and the endosperm does not degrade, suggesting that pressure imposed on the endosperm by embryo growth is required for inducing endosperm cell death (Fourquin et al., 2016). However, the molecular basis for mechanical communication between the embryo and the endosperm, and in particular how mechanical cues in the endosperm are interpreted at the molecular level, remains largely unknown. Chemical communication across the apoplast is traditionally classified as either peptide-based or hormone-based signalling, depending on the biochemical nature of the messenger. Hormones are small molecules of diverse chemical composition. They can be gases, amino acid derivates, nucleotide derivates, etc... The first plant hormone to be described was auxin, a hormone that has subsequently been shown to have an incredible range of physiological effects and to regulate a bewildering array of developmental processes (Darwin, 1881; Tivendale and Cohen, 2015). Seed development, like most other developmental programmes in the plant, is strongly influenced by auxin (Figueiredo and Köhler, 2018; Locascio et al., 2014). In the embryo, auxin is required for the early stages of development as it plays a critical role in patterning the embryonic apical/basal axis (Friml et al., 2003; Robert et al., 2013). This role is the best documented in Arabidopsis (Friml et al., 2003; Jeong et al., 2016; Robert et al., 2013; Smit and Weijers, 2015). Auxin is also massively produced in the endosperm immediately after fertilization, where it triggers endosperm proliferation and differentiation (see our review Doll et al., 2017 in annexe 1) (Batista et al., 2019; Chen et al., 2014; Figueiredo and Köhler, 2018; Figueiredo et al., 2015; Liao et al., 2015). In addition, auxin from the endosperm is thought to be transported to the seed coat just after fertilization where it triggers seed coat development, thus coordinating the development of the fertilized zygotic tissues with that of surrounding maternal tissues (Figueiredo et al., 2016). In addition to auxin, many other hormones are involved in various processes during seed development, including gibberellins, abscisic acid, ethylene and brassinosteroids (Locascio et al., 2014). However, the role of these hormones in embryo/endosperm and growth coordination is poorly understood, partly because of the difficulty of separating their cell autonomous and non-cell autonomous effects. In peptide signalling, the mobile molecule is a peptide, a small protein generally less than one hundred amino acids long. Signalling peptides are generally separated into two categories, small peptides and the cysteine rich peptides. Although signalling peptides are involved in many developmental process, only very few have been proposed to be involved in embryo-endosperm communication (Ingram and Gutierrez-Marcos, 2015; Matsubayashi, 2014). During early seed development in Arabidopsis, three cysteine-rich peptides named EMBRYO SURROUNDING 43 FACTORS (ESF1.1-3) have been found to be mainly expressed in endosperm and have an effect on the development of the suspensor, by controlling suspensor patterning via the SSP and YODA pathway. Exogenous application of ESF1 peptides to developing seeds in vitro has a positive effect on suspensor elongation, showing their capacity to diffusion through seed tissues. In addition, inhibition of ESF1 production specifically in the embryo by RNAi does not lead to a suspensor phenotype, suggesting an non-cell autonomous effect of the peptides (Costa et al., 2014). However, this non-cell autonomous effect is still controversial since transcriptomic data show an expression of ESF1 in the embryo (Bayer et al., 2017). A similar controversy surrounds CLAVATA3/ESR-RELATED 8 (CLE8). CLE8 is a small peptide of the CLE family and has a role in suspensor patterning, through the regulation of WOX8 expression. CLE8 is not expressed in the suspensor suggesting a non-cell autonomous effect, but is expressed in both embryo proper and endosperm. A third case of putative embryo/endosperm peptide-mediated signalling pathway is the previously described GSO1 GSO2 pathway, involved in embryonic cuticle integrity, for which the peptide implicated is still unknown (Xing et al., 2013) (Figure 3B). In maize, no signalling peptide with a confirmed role in embryo/endosperm communication has been described for the moment. However, many peptides are expressed in the tissues at the embryo/endosperm interface, and especially in the ESR (see our review Doll et al., 2017 in annexe )(Zhan et al., 2015). ESR6, a peptide expressed in the ESR was described has having an anti-microbial activity and was proposed to play a role in protecting the embryo from pathogens (Balandín et al., 2005). Among the most strongly expressed genes in the ESR are three CLE peptides. These are amongst the first CLE discovered and are the source of the “E” in the family name “CLE” (Bonello et al., 2000; Cock and McCormick, 2001; OpsahlFerstad et al., 1997). These three secreted peptides have, for the moment, no known function. In the first chapter (section I-2) of this thesis, a particular emphasis will be put on signalling proteins expressed in the ESR cells of maize, using with a reverse genetic approach.
II-3-B) A chemical filter allowing controlled exchanges II-3-B-a) The diffusion of signalling molecules
As discussed in the last subsection, embryo/endosperm communication is likely to require the diffusion of molecules across the embryo/endosperm interface. For example, during the early stages of maize kernel development, the auxin required by the embryo is thought to come from 44 the endosperm and to diffuse into the tip of the embryo proper before being redistributed within the embryo (Chen et al., 2014; Forestan and Varotto, 2012; Forestan et al., 2010)(see our review Doll et al., 2017 annexe 1). In addition to signalling molecules, many other molecules must be taken up by the embryo during embryonic growth. II-3-B-b) The diffusion of nutrients
The embryos of both maize and Arabidopsis not only develop new organs during seed development, but also store large amounts of energy in the form of carbohydrates, lipids and proteins. Embryos thus require various kind of organic and inorganic nutrients for their development and maturation, including diverse ions, sugars and amino acids, for which they are dependent on surrounding tissues. However, the embryo is symplastically isolated, meaning that the nutrient flow transits through the apoplast (Stadler et al., 2005; Van Lammeren, 1987). Two pathways have been suggested to supply embryo needs (Figure 5). The first is via the suspensor, which is in contact with the maternal tissues at its base. Maternal nutrients could reach directly the embryo without crossing the endosperm. However, as the suspensor is mainly surrounded by endosperm, and expresses numerous genes encoding nutrient transporters it has been proposed to predominantly act as an organ that takes up nutrients from the endosperm (Kawashima and Goldberg, 2010). The suspensor is relatively large compared to the embryo proper during the first stages of embryogenesis, and has a much larger surface area. However as the embryo expands this relationship inverses and the suspensor degenerates (Blanvillain et al., 2011; Giuliani et al., 2002). As a consequence of this degeneration, the suspensor pathway is probably replaced by a second pathway via which the embryo can take up nutrients from the endosperm although, surprisingly, almost nothing is known about this process (Figure 5) (Karmann et al., 2018; Lafon-Placette and Köhler, 2014). Apoplastic nutrient transport is facilitated by transmembrane transporters that export molecules from source tissues and import them in the sink tissue. Many transmembrane transporters have been shown to be expressed on both side of embryo/endosperm interface in Arabidopsis (Figure 5). The bidirectional sugar transporters SWEET 11 and 15, as well as the sugar and biotin transporter AtSUC5 are expressed in the endosperm cells surrounding the embryo and are involved in the embryo filling (Baud et al., 2005; Chen et al., 2015; Pommerrenig et al., 2013). Some amino acid transporters are also expressed in the endosperm, but not specifically in the cells surrounding the embryo. These include AtCAT6, UMAMIT25 and 28. However, the role of these proteins in embryo filling has not yet been demonstrated (Besnard et al., 2018; Hammes et al., 2006; Müller et al., 2015). At the embryo side, sugars could be taken up by AtSUC3 or SWEET12 which are
45 Figure 5: Genes encoding sugar and amino acid transporters expressed on either side of the embryo/endosperm interface in Arabidopsis. The arrows represent the putative main pathways for nutrient loading to the embryo. In purple via the suspensor and in red via the epidermis. 46 expressed in the suspensor at early developmental stages (Chen et al., 2015; Stadler et al., 2005).
At
later
stages
, AtSUC5 is express
ed
in
the epidermis of the embryo proper, and the
AMINO ACID
TRANSPORTER (AAP
1) gene is expressed in the whole embryo proper (Pommerrenig et al., 2013; Sanders et al., 2009). For AAP1, a role in amino acid uptake by the embryo has been demonstrated, strengthening the evidence for a second transport pathway from the endosperm (Sanders et al., 2009). Other nutrients, such as iron, phosphate and nitrate are also transported to the embryo, although detailed studies of transporters localisation are globally lacking (Curie and Mari, 2017; Tsay et al., 2007; Vogiatzaki et al., 2017). In maize very little is known about the nutrient flow transiting through the endosperm/embryo interface or about the transporters expressed at the interface, despite the evident potential agronomic interest. Transporter-encoding genes expressed at the embryo/endosperm interface will be investigated further using transcriptomic approaches in the first chapter of this thesis. II-3-B-c) The diffusion barrier, a filtering role?
Although nutrients and small molecules diffuse into the embryo from the endosperm, the permeability of the interface is thought to be controlled by apoplastic barriers. In maize the diffusion of small aqueous and ethanol dyes at the embryo/endosperm interface is reduced at maturity compared to the diffusion inside either the endosperm or the scutellum (CoronaCarrillo et al., 2014). In Arabidopsis, a diffusion barrier (the embryonic cuticle) is built up around the embryo proper during seed development (Creff et al., 2019). Consistent with this, the permeability of the embryo surface, revealed by the penetration of a small hydrophilic dye called toluidine blue, reduces during embryo development suggesting a decrease in barrier permeability (Creff et al., 2019) (Figure 6). Interestingly, permeability is retained at the root pole of the embryo, despite the presence of a cuticle like barrier, making it possible that nutrients could diffuse into the embryo in this zone (Berhin et al., 2019). Moreover, for the suspensor, which is devoid of cuticle, permeability is thought to be very high (Kawashima and Goldberg, 2010; Szczuka and Szczuka, 2003). Mutants defective in cuticle integrity show higher embryonic permeability to toluidine blue, suggesting a key role of the cuticle in limiting the diffusion at the surface of the embryo proper (Creff et al., 2019; Delude et al., 2016; Moussu et al., 2013). As permeability to toluidine blue is only very weakly affected in krs, and likely only after germination, the contribution of the sheath to embryo impermeability to small hydrophilic molecules appears not to be very important. However, permeability was only assessed with toluidine blue and the permeability to other molecules could be different (Tanaka 47 A B Toluidine blue
Epidermal cells Seedling Intact cuticle Defective cuticle
Figure 6: A) Toluidine blue penetration in dissected embryos at different developmental stages compared to unstained embryos (bottom line). (Taken from Creff et al., 2019, figure made by the candidate). Scale bars = 100μm. B) Scheme representing the toluidine blue staining assay used for revealing defects in the cuticle. 48 et al., 2004). Diffusion to the embryo could be restricted by the retention of molecules in the apoplast, which could be particularly relevant for the embryo sheath since EXTENSINS are thought to be highly electronegative. Limiting diffusion at the embryo surface would lead to the formation of chemically separated compartments. One hypothesis regarding the role of this isolation is that it could protect the embryo from cell wall degrading enzymes expressed in the endosperm, or, alternatively from molecules produced during endosperm cell death (Moussu, 2016). More globally, these barriers could control the mobility of molecules diffusing into the embryo. Interestingly in maize, the diffusion barrier may not only be restricted to apoplastic structures but could involve an entire tissue. The maize ESR has been propose to limit auxin diffusion from the surrounding endosperm and to protect the embryo from high auxin concentrations by actively taking up and sequestering auxin (Chen et al., 2014)(see our review Doll et al., 2017 in annexe 1).
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Secrecy Outage Analysis for Downlink Transmissions in the Presence of Randomly Located Eavesdroppers
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Gaojie Chen
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To cite this version: Gaojie Chen, Justin P Coon, Marco Di Renzo. Secrecy Outage Analysis for Downlink Transmissions
in the Presence of Randomly Located Eavesdroppers. IEEE Transactions on Information Forensics
and Security, 2017, 12 (5), pp.1195 - 1206. 10.1109/TIFS.2017.2656462. hal-01880018 Distributed under a Creative Commons Attribution 4.0 International License Secrecy Outage Analysis for Downlink
Transmissions in the Presence of
Randomly Located Eavesdroppers e Chen, Member, IEEE, Justin P. Coon, Senior Member, IEEE, and Marco Di Renzo, Se at a rate close to the intended channel capacity so that only
the intended receiver can successfully decode the data. This
is the principle of physical layer security, where the level
of security is quantified by the secrecy capacity, i.e., the
difference in channel capacities corresponding to the intended
data transmission and EDs. at a rate close to the intended channel capacity so that only
the intended receiver can successfully decode the data. This
is the principle of physical layer security, where the level
of security is quantified by the secrecy capacity, i.e., the
difference in channel capacities corresponding to the intended
data transmission and EDs. Abstract—We analyze the secrecy outage probability in the
downlink for wireless networks with spatially (Poisson) distrib-
uted eavesdroppers (EDs) under the assumption that the base
station employs transmit antenna selection (TAS) to enhance
secrecy performance. We compare the cases, where the receiving
user equipment (UE) operates in half-duplex (HD) mode and
full-duplex (FD) mode. In the latter case, the UE simultaneously
receives the intended downlink message and transmits a jam-
ming signal to strengthen secrecy. We investigate two models
of (semi)passive eavesdropping: 1) EDs act independently and
2) EDs collude to intercept the transmitted message. For both
of these models, we obtain expressions for the secrecy outage
probability in the downlink for the HD and FD UE operation. The
expressions for the HD systems have very accurate approximate
or exact forms in terms of elementary and/or special functions
for all path loss exponents. Those related to the FD systems
have exact integral forms for general path loss exponents, while
exact closed forms are given for specific exponents. A closed-form
approximation is also derived for the FD case with colluding
EDs. The resulting analysis shows that the reduction in the
secrecy outage probability is logarithmic in the number of
antennas used for TAS and identifies conditions, under which
HD operation should be used instead of FD jamming at the UE. These performance trends and exact relations between system
parameters can be used to develop adaptive power allocation
and duplex operation methods in practice. Examples of such
techniques are alluded to herein. HAL Id: hal-01880018
https://hal.science/hal-01880018v1
Submitted on 16 Jul 2020 L’archive ouverte pluridisciplinaire HAL, est
destinée au dépôt et à la diffusion de documents
scientifiques de niveau recherche, publiés ou non,
émanant des établissements d’enseignement et de
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publics ou privés. HAL is a multi-disciplinary open access
archive for the deposit and dissemination of sci-
entific research documents, whether they are pub-
lished or not. The documents may come from
teaching and research institutions in France or
abroad, or from public or private research centers. Distributed under a Creative Commons Attribution 4.0 International License 1195 IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 Manuscript received August 9, 2016; revised November 4, 2016; accepted
December 27, 2016. Date of publication January 20, 2017; date of current
version February 22, 2017. This work was supported by EPSRC under
Grant EP/N002350/1 (“Spatially Embedded Networks”). The associate editor
coordinating the review of this manuscript and approving it for publication
was Dr. Liang Xiao. Secrecy Outage Analysis for Downlink
Transmissions in the Presence of
Randomly Located Eavesdroppers Recently, many works have considered information theo-
retic security (ITS) over wireless channels, including coop-
erative relay and jammer networks [2], [3], buffer-added
relay networks [4], multiple-input multiple-output communi-
cations (MIMO) [5], [6], full-duplex networks [7], cognitive
radio networks [8], and distributed beamforming methods [9]. However, all of these works not only assumed a small number
of nodes, but also assumed the locations of EDs are known. It is impossible to obtain the location of EDs in practice. For
this reason, in 2008, Haenggi provided a powerful method to
model the random location distribution of nodes in wireless
networks [10], [11]. The impact of random ED locations on secrecy performance
has been investigated [12]–[16]. The location distribution of
EDs can be modeled as a Poisson point process (PPP) or a
binomial point process (BPP). In [12], the locations of multiple
legitimate pairs and EDs were represented as independent two-
dimensional PPPs, and the average secrecy throughput in such
a wireless network was studied. The MIMO transmission with
beamforming was considered later in [13] and [14] to enhance
secrecy performance. Index Terms—Physical layer security, stochastic geometry,
secrecy outage probability, antenna selection, full-duplex. Manuscript received August 9, 2016; revised November 4, 2016; accepted
December 27, 2016. Date of publication January 20, 2017; date of current
version February 22, 2017. This work was supported by EPSRC under
Grant EP/N002350/1 (“Spatially Embedded Networks”). The associate editor
coordinating the review of this manuscript and approving it for publication
was Dr. Liang Xiao.
G. Chen and J. P. Coon are with the Department of Engineering
Science,
University
of
Oxford,
Oxford
OX1
3PJ,
U.K.
(e-mail:
gaojie.chen@eng.ox.ac.uk; justin.coon@eng.ox.ac.uk).
M. Di Renzo is with the Laboratory of Signals and Systems, University of
Paris-Sud, 91400 Orsay, France (e-mail: marco.direnzo@lss.supelec.fr).
Color versions of one or more of the figures in this paper are available
online at http://ieeexplore.ieee.org.
Digital Object Identifier 10.1109/TIFS.2017.2656462 This work is licensed under a Creative Commons Attribution 3.0 License. For more information, see http://creativecom er a Creative Commons Attribution 3.0 License. For more information, see http://creativecommons.org/licenses/by/3.0/ G. Chen and J. P. Coon are with the Department of Engineering
Science,
University
of
Oxford,
Oxford
OX1
3PJ,
U.K.
(e-mail:
gaojie.chen@eng.ox.ac.uk; justin.coon@eng.ox.ac.uk). Digital Object Identifier 10.1109/TIFS.2017.2656462 This work is licensed under a Creative Commons Attribution 3.0 License. For more information, see http://creativecommons.org/licenses/by/3.0/ A. System Model We consider a secure transmission from the BS to one
legitimate UE.1 The BS is equipped with K antennas, which it
uses to perform TAS in order to maximize the instantaneous
signal-to-noise ratio (SNR) at the UE. The UE is equipped
with a hyper-duplex antenna, which can easily switch between
HD and FD modes. Without loss of generality, we locate the
BS at the origin in R2 and locate the UE at a fixed point a
distance dBU along the positive x-axis (see Fig. 1). distributed EDs. In order to keep the complexity relatively
low at the base station (BS), we consider transmit antenna
selection (TAS) rather than beamforming. Furthermore, we
compare the cases where the receiving user equipment (UE)
operates in half-duplex (HD) mode and full-duplex (FD) mode. In the latter case, the UE simultaneously receives the intended
downlink message and transmits a jamming signal to disrupt
eavesdropping devices [7]. We also treat the case when EDs act
independently as well as the scenario when they collude. The
analytical framework that we present in this paper allows us to
make a fair comparison of these four system models (HD/FD
and independent/colluding EDs) and thus to draw conclusions
about the relative merits and drawbacks of using the secrecy
enhancement techniques of TAS and FD jamming under given
system parameterizations. The contributions of the paper are
summarized as follows. We assume EDs are randomly dispersed in a region in the
neighbourhood of the BS and the UE. To this end, we model
the EDs as a PPP , which has intensity ρE in the closed disk
of radius R, which we denote by V, centred at the origin and
zero intensity in R2 \ V (Fig. 1). Each ED is equipped with
a single antenna, but we consider both the scenarios in which
EDs attempt to intercept the downlink signal independently as
well as the case when EDs collude to decode the transmitted
message. g
All channels are assumed to undergo path loss and indepen-
dent Rayleigh fading effects. Hence, the coefficient modeling
the channel between nodes i and j can be decomposed
as gij = hij d−α/2
ij
, where α and dij denote the path loss
exponent and the distance between the two nodes, respec-
tively.2 The fading coefficient hij is modeled as a complex
Gaussian random variable with unit variance (i.e., Rayleigh
fading is assumed). TABLE I
NOTATION AND SYMBOLS USED IN THE PAPER TABLE I Fig. 1. The wireless network model with randomly located EDs and fixed
BS and UE. 1If there are several users in the target cell, only one user is targeted through
user scheduling (e.g. random user selection).
2In what follows, we set the subscripts i and j to be elements in the
set {B,U, E} in order to denote transmissions from the BS, UE and EDs,
respectively. For example, gU E1 denotes the channel coefficient between the
UE and the first ED in . I. INTRODUCTION P P
HYSICAL layer security, based on Shannon theory using
channel coding to achieve secure transmission, has been
frequently considered in academia since Wyner’s seminal
work [1]. Due to the broadcast nature of wireless commu-
nications, both the intended receiver and eavesdroppers (EDs)
may receive data from the source. But if the capacity of
the intended data transmission channel is higher than that
of the eavesdropping channel, the data can be transmitted Cooperation is of paramount importance to enhance the
capacity and reduce the outage of communication systems
subjected to fading and unknown topologies [17]. As a result,
cooperation schemes have been widely applied to enhance
communication between legitimate users in a physical layer
secrecy context [2], [3]. However, relatively little attention
has been given to the impact of colluding or cooperative
EDs in random spatial networks. Notably, [18] investigated
achievable secrecy rates by using the so-called intrinsically
secure graph formalism, taking into account the effects of ED
collusion. Additionally, based on a beamforming technique,
the MIMO secrecy connectivity between devices operating
in the presence of Rayleigh fading and colluding EDs was
analysed in [19]. However, in that work, the complexity of
the system is high due to the use of multiple antennas with
beamforming, which may render the system unsuitable for
some practical applications. In this paper, we analyze the secrecy outage probability in
the downlink for wireless networks with randomly (Poisson) IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 1196 Fig. 1. The wireless network model with randomly located EDs and fixed
BS and UE. TABLE I
NOTATION AND SYMBOLS USED IN THE PAPER 2In what follows, we set the subscripts i and j to be elements in the
set {B,U, E} in order to denote transmissions from the BS, UE and EDs,
respectively. For example, gU E1 denotes the channel coefficient between the
UE and the first ED in . 1If there are several users in the target cell, only one user is targeted through
user scheduling (e.g. random user selection). B. Secrecy Performance and F(·) is an operator that takes different forms depending
on whether EDs act independently or whether they collude. In the former case, we have We define downlink secrecy performance using classical
wireless wiretap theory. We assume the channel state informa-
tion (CSI) between the BS and the UE is known by the BS.3
Therefore, by employing the TAS principle, the BS is able
to send a zero-mean symbol xs with E[|xs|2] = 1 to the UE
by selecting the kth antenna (corresponding to the maximum
instantaneous downlink SNR) in a given time slot. F(·) = max
e∈ (·)
(8) (8) Therefore, by employing the TAS principle, the BS is able
to send a zero-mean symbol xs with E[|xs|2] = 1 to the UE
by selecting the kth antenna (corresponding to the maximum
instantaneous downlink SNR) in a given time slot. so that we ensure we consider the strongest ED channel,
whereas in the case of colluding eavesdroppers, the operator
is given by so that we ensure we consider the strongest ED channel,
whereas in the case of colluding eavesdroppers, the operator
is given by is given by g
In general, the received signal at the UE can be written
as In general, the received signal at the UE can be written
as F(·) =
e∈
(·)
(9) (9) as yBkU =
PBgBkU xs + ϖ
PU gUUx j + nU
(1) (1) since all EDs are capable of combining their signals in
an optimal manner to decode the message. Based on these
formulae, the secrecy outage probability can be defined as [20] where PB is the average transmit power at the BS and
nU denotes zero-mean complex Gaussian noise with vari-
ance σ 2
n . The coefficient gUU corresponds to the residual
self-interference channel for the case where FD jamming is
employed and x j denotes the zero-mean jamming signal which
has power E[|x j|2] = 1. The average transmit power of the
FD UE is PU. Eq. (1) can be applied to model systems with
both HD and FD UEs by adjusting the parameter ϖ. In the
HD case, ϖ = 0, whereas in the FD case, ϖ = 1. B. Secrecy Performance Pso = P([CBU −CBE∗]+ < ϵ) ≃P
γBU
γBE∗
< β
(10) (10) where [x]+ = max(0, x), P(·) denotes the probability operator,
ϵ denotes the target secrecy rate, β = 2ϵ denotes the target
secrecy SNR ratio.4 4The approximation in (10) is a standard assumption for systems operating
in the high SNR region. In this paper, this condition implies PB is sufficiently
large and/or R is sufficiently small. CHEN et al.: SECRECY OUTAGE ANALYSIS FOR DOWNLINK TRANSMISSIONS IN THE PRESENCE OF RANDOMLY LOCATED EDs
1197 CHEN et al.: SECRECY OUTAGE ANALYSIS FOR DOWNLINK TRANSMISSIONS IN THE PRESENCE OF RANDOMLY LOCATED EDs
1197 1197 III. SECRECY OUTAGE PROBABILITY FOR
INDEPENDENTLY ACTING EAVESDROPPERS At the same time that the UE receives the message from
the BS, the EDs in the set receive a copy of the transmitted
signal. The received signal at ED Ee can be written as Here, we analyse the secrecy outage probability of the
downlink for HD and FD UEs under the assumption that EDs
act independently of one another. The EDs cannot share their
received signals in this case, so secrecy outage is dictated by
the ED with highest channel capacity. Hence, F(·) is defined
by (8). We begin by considering an HD UE, then proceed with
a treatment of the problem for an FD UE. yBkEe =
PBgBkEexs + ϖ
PU gU Eex j + nEe
(2) (2) where nEe is the Gaussian noise (with variance σ 2
n ) at the ED. We are interested in quantifying the secrecy outage
probability in the downlink. To this end, we require expres-
sions for the BS-UE and BS-ED channel capacities. Based
on the models described above, the capacity of the BS-UE
channel can be written as A. System Model Therefore, the corresponding channel
gains |gij|2 are independently exponentially distributed with
mean value λij , and the average channel power is given by
λij = E[|gij|2] = d−α
ij , where E[·] denotes the expectation
operation. We assume that the channels are quasi-static, so
that the channel coefficients remain unchanged during several
packet transmissions but independently vary from coherence
time interval to another. • We propose TAS at the BS and FD jamming at the
receiver to enhance secrecy performance in the presence
of randomly located EDs. • We obtain expressions for the secrecy outage probability
in the downlink for HD and FD receivers operating in the
presence of independent and colluding EDs. The expres-
sions for HD systems have very accurate approximate
or exact forms in terms of elementary and/or special
functions for all path loss exponents. Those related to the
FD systems have exact integral forms; exact closed forms
are given for certain path loss exponents and closed-form
approximations are also derived. The remainder of the paper is organized as follows. Section II presents the system model and problem formulation. Sections III and IV given an analysis of the secrecy outage
probability for the cases where EDs act independently and
when they collude, respectively. Section V gives numerical
simulations in order to verify the analysis. Finally, section VI
concludes the paper. The notation and symbols used in the
paper are listed in Table I. A. Half Duplex UE Beginning with the right-hand side of (10), the secrecy
outage probability can be evaluated to yield the result stated
in the following proposition. CBU = log2(1 + γBU)
(3) (3) Proposition 1: For large R, the downlink secrecy outage
probability for an HD UE is, to a good approximation, given
by where where γBU =
PB max
k∈{1...K}
|hBkU |2
dα
BU
ϖ PU|gUU|2 + σ 2n
(4) P(H)
so
≃1 −
K
k=1
(−1)k+1Ck
K
√pq
2
p+2q−3
2
π
p+2q
2
−1
× G p+2q,0
0,p+2q
a2q
k b p
p p4qq2q
−
0, 1
p, ..., p−1
p , 1
2q , 2
2q , ..., 1
(11) (4) and the max operation results from the TAS scheme at the BS. For the BS-ED channel, the capacity is given by and the max operation results from the TAS scheme at the BS. For the BS-ED channel, the capacity is given by CBE∗= log2(1 + γBE∗)
(5) CBE∗= log2(1 + γBE∗) (5) (5) (5)
(11) where where Gm,n
s,t
z
u1, . . . , us
v1, . . . , vt
is the Meijer G function, Ck
K = where γBE∗= F
⎛
⎜⎝
PB|hB∗Ee |2
dα
BEe
ϖ PU |hU Ee |2
dα
U Ee
+ σ 2n
⎞
⎟⎠
(6)
B∗= arg
max
k∈{1...K}
|h BkU|2
dα
BU
(7) γBE∗= F
⎛
⎜⎝
PB|hB∗Ee |2
dα
BEe
ϖ PU |hU Ee |2
dα
U Ee
+ σ 2n
⎞
⎟⎠
1,
,
t
K!/((K −k)!k!) is the binomial coefficient, ak = kdα
BU, b =
πρE (1 + 2/α)β2/α, p, q ∈Z+ so that α = p/q is a positive
rational number, and (x) =
∞
0
tx−1et dt is the standard
gamma function. (6) (6) with Proof: See Appendix I. g
Proof: See Appendix I. B. Full Duplex UE In the case where FD jamming is employed by the UE,
the jamming signal will affect both the EDs and the UE. Thus, a self-interference cancellation scheme must be applied
at the UE. Here, we assume the self-interference cancellation
scheme is not perfect, and thus residual interference will
remain. Also, we are interested in the worst-case secrecy
performance. Thus, in this section, we assume the EDs
are interference limited (from the UE’s jamming signal). Mathematically, we set σ 2
n = 0. A similar approach was taken
in [21]–[23]. Now, beginning with the right-hand side of (10),
the secrecy outage probability can be evaluated to yield the
result stated in the following proposition. P(H)
so
≃1 −2
K
k=1
(−1)k+1Ck
K
akb K1
2
akb
(12) However, for other values of α, the expression given in
the proposition is the most compact, accessible form. Note
that the expression given in Proposition 1 is independent
of R. This is because the R-dependent terms in the secrecy
outage probability expression decay exponentially with Rα. (See Appendix I for details.) Proposition 2: The downlink secrecy outage probability for
an FD UE located in the presence of independently acting EDs
is upper bounded by P(F)
so
≤1 −e−ρE π R2
K
k=1
(−1)k+1kCk
K
×
∞
0
PU
dα
BU (1 + λUU) + kxλUU
( PU
dα
BU + kxλUU)2
× exp
ρE R2
x
β ; α, dBU
R
−kdα
BU
PU
x
dx
(14)
where P(F)
so
≤1 −e−ρE π R2
K
k=1
(−1)k+1kCk
K For fixed dBU, ρE, β, and α, the secrecy outage probability
solely depends on the available number of BS antennas K. It is not a function of the transmit power PB. This is perfectly
intuitive since an increase in PB would yield a proportional
increase in both the UE SNR and the ED SNR. Thus, in order
to satisfy a given secrecy requirement, one must increase the
number of antennas used in the TAS procedure. With large-
scale antenna systems and massive MIMO making headlines
in the research community in recent years, it is prudent to ask
how the secrecy outage probability scales with the number of
antennas used for selection. Since the BS-ED channels are not
considered in the selection process, it is clear that the secrecy
outage probability decreased monotonically with increasing K. Proof: See Appendix I. B∗= arg
max
k∈{1...K}
|h BkU|2
dα
BU
(7) Eq. (11) provides an explicit, relation between the secrecy
outage probability and various system parameters. A number (7) 3This can be achieved by feeding back CSI from the UE to the BS directly or
through channel reciprocity in the case of time-division duplex transmissions. IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 1198 The actual benefit brought by TAS in the context of enhancing
secrecy performance is explored further in Section V through
numerical simulations. of interesting points can be noted from this expression. First,
this is the most complete analysis of the HD UE case reported
in the literature in that any rational path loss exponent is
accounted for in this expression. Indeed, since the path loss
exponent is an experimentally estimated parameter, it is, by
definition, rational in practice due to finite precision measure-
ment equipment. Although the outage probability is given in
terms of the Meijer G function, it can be easily evaluated
using numerical software such as Mathematica or Maple for
any given inputs. It should be noted that for the special case
of α = 2, (11) reduces to the following expression written in
terms of first order modified Bessel functions of the second
kind: Proof: See Appendix III. Proof: See Appendix III. Lemma 1: The downlink secrecy outage probability for an
HD UE located in the presence of independently acting EDs
is lower bounded by The bound stated above can be evaluated for given sets of
parameters by using standard numerical integration techniques
or software. Note that the semi-infinite integral is guaranteed to
converge since
(y; α, δ) is finite for y ∈[0, ∞). For the case
where α = 2, the bound simplifies somewhat since
(y; α, δ)
evaluates to P(H)
so
> πρEd2
BUβ2/α (1 + 2/α)
e (ln K)2/α
1 + O
1
(ln K)2/α
(13) P(H)
so
> πρEd2
BUβ2/α (1 + 2/α)
e (ln K)2/α
1 + O
1
(ln K)2/α
(13) (y; 2, δ)
=
πy
(y + 1)3
(y + 1)(ψ(y, δ) −δ2)
+ δ2(y −1) ln
2δ2y
δ2(y −1) + (y + 1)(ψ(y, δ) + y + 1)
(16) as K →∞. Proof: See Appendix II. B. Full Duplex UE But how fast does this occur? The following lemma provides
some insight to this question.
0
( PU
dα
BU + kxλUU)2
× exp
ρE R2
x
β ; α, dBU
R
−kdα
BU
PU
x
dx
(14) where where
(y; α, δ) =
2π
0
1
0
yzα+1
yzα + (z2 + δ2 −2zδ cos θ)α/2 dz dθ
(15) (15) (15) and λUU
= E[|gUU|2] is the average gain of the self-
interference channel at the FD UE. and λUU
= E[|gUU|2] is the average gain of the self-
interference channel at the FD UE. B. Full Duplex UE degrees of freedom that can be considered at a system level
when determining the best configuration for achieving a target
secrecy outage probability. For example, the UE may locally
determine that it should reduce PU to conserve battery power,
which implies the BS should increase the number of antennas
used for TAS. Further analysis of the trade-off between these
parameters and the effect this has on system performance are
presented in Section V. When FD jamming is utilized by the UE, we assume self-
interference cancellation is employed by the UE and consider
the interference limited regime for EDs (i.e., σ 2
n
= 0 at
each ED). Following from the right-hand side of (10), the
secrecy outage probability in this scenario can be evaluated to
yield the tight bound stated in the following proposition. Proposition 4: The downlink secrecy outage probability for
an FD UE located in the presence of colluding EDs is bounded
by A. Half Duplex UE By using the right-hand side of (10) the secrecy outage
probability can written exactly as in Proposition 3. (22) Proof: See Appendix V. Proof: See Appendix V. Proposition 3: The downlink secrecy outage probability for
an HD UE located in the presence of colluding EDs is given
by Eq. (21) can be evaluated for given sets of parameters by
using standard numerical integration techniques or software. Eq. (21) can be evaluated for given sets of parameters by
using standard numerical integration techniques or software. However, it is useful to have an approximation of this expres-
sion that does not require numerical integration. We give such
an approximation for α = 2 in the following lemma, and
we validate the approximation in the next section through an
extensive simulation study. P(H)
so
= 1 −
K
k=1
Ck
K (−1)k+1
× exp
−π R2ρE F
1, 2
α ; 1 + 2
α ; −
Rα
kβdα
BU
(18 Lemma 2: For α = 2, the downlink secrecy outage proba-
bility for an FD UE located in the presence of colluding EDs
operating in the interference limited regime is approximated
by (18) where F(a, b; c; z) denotes the Gaussian hypergeometric
function. by P(F)
so
≃1 +
K
k=1
Ck
K (−1)k exp
−ρE
πϱ2
−π PU
2kβ
(ϱ/dBU)2 −ln(1 −(ϱ/dBU)2)
+ (β; dBU, R, A0)
(23) Proof: See Appendix IV. Proof: See Appendix IV. Eq. (18) provides an explicit, exact relation between the
secrecy outage probability and various system parameters. For
α = 2, this expression simplifies readily to P(H)
so
= 1 −
K
k=1
Ck
K (−1)k+1
1 +
R2
βd2
BUk
−πρE βd2
BU k
. (23) (19)
where ϱ ∈(0, R), A0 = 2kβd2
BU/PU, and (β; dBU, R, A0)
is given as (24) at the bottom of the next page. (19) (19) For α = 4, (18) can be expressed as For α = 4, (18) can be expressed as For α = 4, (18) can be expressed as Proof: See Appendix V. P(H)
so
= 1 −
K
k=1
Ck
K (−1)k+1
× exp
−πρE RdBU
βk tan−1
R
dBU
√βk
. (20) IV. SECRECY OUTAGE PROBABILITY FOR
COLLUDING EDs P(F)
so
≤1 +
K
k=1
Ck
K (−1)k
× exp
−ρE
R
0
2π
0
Ak(r, θ)eAk(r,θ)E1
(Ak(r, θ))r dθ dr
,
(21) Here, we analyse the secrecy outage probability in the
downlink for HD and FD UEs with the assumption that
EDs collude with each other. In contrast to independently
acting EDs, colluding EDs can share their eavesdropping
information; therefore, all the eavesdropping information can
be combined in an effort to decode the downlink message. Under the assumption that optimal combining can be achieved
by the EDs, F(·) is defined by (9). We first consider an HD
UE, then a treatment of the problem for an FD UE will be
provided. (21) where E1(x) =
∞
x
e−t
t dt denotes the exponential integral and Ak(r, θ) = 2kβ
PU
dα
BU
⎛
⎝
r
r2 + d2
BU −2rdBUcos(θ)
⎞
⎠
−α
. CHEN et al.: SECRECY OUTAGE ANALYSIS FOR DOWNLINK TRANSMISSIONS IN THE PRESENCE OF RANDOMLY LOCATED EDs CHEN et al.: SECRECY OUTAGE ANALYSIS FOR DOWNLINK TRANSMISSIONS IN THE PRESENCE OF RANDOMLY LOCAT 1199 as K →∞. Proof: See Appendix II. This result implies that, for large numbers of antennas,
secrecy performance improves slowly with increasing K. From a system design perspective, this is a very important
result. It suggests that even systems with large numbers of
antennas (e.g., massive MIMO systems with a TAS-based
secrecy enhancement mode) should exploit only a small subset
of independent spatial paths to perform selection. Such an
approach would allow the remaining elements to serve other
UEs on separate channels. The total number of transmit chains
(i.e., up-conversion and power amplification circuitry) required
would be the number of UEs served in a single channel use. where ψ(y, δ) =
δ4 + 2δ2(y −1) + (y + 1)2. (17) (17) For fixed dBU, ρE, λUU, β, and α, the secrecy outage prob-
ability depends on the available number of BS antennas K, but
also on the UE jamming signal power PU. This provides two V. SIMULATIONS RESULTS In this section, simulation results (based on the left-hand
side of (10)) are given to verify the above analysis. In the
simulations, we assume the noise variance σ 2
n
= 1, the
transmission-power-to-noise ratio PB/σ 2
n = 50 dB, and the
target secrecy SNR β = 1. The simulation results are obtained
by averaging over 105 independent Monte Carlo trials. More-
over, the single-antenna scheme (K = 1) is our benchmark
and has been considered in this section. (20) Other values of the path loss exponent do admit closed
form expressions by eq. (18). To avoid the redundant
discussion, we have not mentioned another pathloss exponents
here. IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 1200 Fig. 2. Theoretical (T.R.) vs simulated (S.R.) secrecy outage probabilities
for the HD UE in the presence of different densities of EDs, where α = 4,
dBU = 10 m and R = 100 m. TABLE II
EFFECTS OF PARAMETERS INCREASES ON SECRECY OUTAGE PROB-
ABILITY. UPWARD (DOWNWARD) ARROWS SIGNIFY AN INCREASE
(DECREASE). HORIZONTAL DASHES DENOTE LITTLE
TO
NO
CHANGE. AN ARROW FOLLOWED BY A DASH SIGNIFIES CON-
VERGENCE TO A POSITIVE, FINITE VALUE. ARROWS FOL-
LOWED BY PARENTHETICAL EXPRESSIONS DENOTE THE
TREND OF INCREASE/DECREASE (EITHER LOGA-
RITHMIC OR A POWER LAW) TABLE II
EFFECTS OF PARAMETERS INCREASES ON SECRECY OUTAGE PROB-
ABILITY. UPWARD (DOWNWARD) ARROWS SIGNIFY AN INCREASE
(DECREASE). HORIZONTAL DASHES DENOTE LITTLE
TO
NO
CHANGE. AN ARROW FOLLOWED BY A DASH SIGNIFIES CON-
VERGENCE TO A POSITIVE, FINITE VALUE. ARROWS FOL-
LOWED BY PARENTHETICAL EXPRESSIONS DENOTE THE
TREND OF INCREASE/DECREASE (EITHER LOGA-
RITHMIC OR A POWER LAW) TABLE II Fig. 2. Theoretical (T.R.) vs simulated (S.R.) secrecy outage probabilities
for the HD UE in the presence of different densities of EDs, where α = 4,
dBU = 10 m and R = 100 m. Firstly, Table II gives an overview of how different system
parameters affect secrecy outage for the four cases discussed
in the previous sections, where IE and CE denote indepen-
dent and colluding eavesdropper case, respectively, ↗, ↘
and −denote increasing, decreasing and unchanging trends,
respectively. It is clear that the secrecy outage probability for
each of the four cases increases with increasing ED density,
target SNR β, and BS-UE distance dBU. V. SIMULATIONS RESULTS On the contrary,
the secrecy outage probability decreases with the number of
transmission antennas K. With the increasing of α, the secrecy
outage probability in the HD case decreases slowly while the
secrecy outage probability in the FD case increases steadily
until it converges to a finite value (more details in Fig. 7). Note that the secrecy outage probability is independent of
the transmit power-to-noise ratio PB/σ 2
n for the BS. Finally,
the transmit power-to-noise ratio PU/σ 2
n for the UE and the
residual self-interference channel gain (λUU) only affects the
FD case which is shown in Figs. 5 and 6, respectively. Fig. 3. The comparison of secrecy outage probabilities for HD UEs with
different numbers of antennas (K), where ρE = 0.005 m−2, α = 2 dBU =
5 m and R = 50 m. Fig. 3. The comparison of secrecy outage probabilities for HD UEs with
different numbers of antennas (K), where ρE = 0.005 m−2, α = 2 dBU =
5 m and R = 50 m. Fig. 2
verifies
the
secrecy
outage
probabilities
for
the HD UE for independent EDs (11) and colluding EDs (18),
respectively. Here we let dBU = 10 m, R = 100 m and
α = 4. Both the simulated results (S.R.) and theoretical
results (T.R.) are presented, which are shown to perfectly
match. Furthermore, it is clear from these results that the
secrecy outage probability slowly decreases as the number
of transmit antennas increases for both cases, which has
been predicted by Lemma 2. The secrecy outage proba-
bility for independent EDs is always smaller than that for
the colluding case, because of the shared eavesdropping
information. Fig. 3 compares secrecy outage probabilities for HD
UEs
with
different
numbers
of
antennas
(K),
where
ρE = 0.005 m−2, α = 2 dBU = 5 m and R = 50 m. It is clear to see that when the number of antennas ranges
from 1 to 15, there exists a significant secrecy performance
gain. However, with increasing numbers of antennas after 15,
secrecy performance improves slowly with 1/ln(K), which
has been confirmed by Lemma 2. From a system design
perspective, this is a very important result. V. SIMULATIONS RESULTS Therefore, we should increase the jamming power PU accord-
ing to the theoretical expressions given in Propositions 3 and 5
so that the secrecy outage probability can be reduced. This
information can be employed in practice to switch between
HD and FD modes given the bandwidth constraints of the
system with different path loss exponents. Since the available
system bandwidth of modern communication links can change
based on channel quality and the prescribed quality of service,
this observation could be of great importance in future cellular
networks [24]. The comparison between the T.R. and S.R. of secrecy outage
probabilities for the FD UE is shown in Fig. 4, where we
let λUU = 0 dB, dBU = 5 m, R = 50 m, ϱ = 15 and
α = 2. Again, the theoretical results generated with the
help of (14) for independent EDs and (21) for colluding
EDs are well matched to the simulation results. And the
approximation results (A.R.) (23) for colluding EDs were
confirmed by simulation results as well. Moreover, it is clear
that the secrecy outage probability decreases exponentially
quickly as the density of EDs decreases, as predicted by
Propositions 3 and 5. Fig. 5 shows the comparison between the T.R. and S.R. of secrecy outage probabilities versus different transmission
power-to-noise ratio for the FD UE in the presence of inde-
pendent and colluding EDs, where λUU = 0 dB, dBU = 5 m,
R = 50 m, ρE = 0.005 m−2 and α = 2. For these system
parameters, the average number of EDs located in the vicinity
of the BS (i.e., the circle of radius R centered at the BS)
is approximately 39. Hence, these parameters provide a view
of performance in a fairly hostile environment. We can see
that the T.R. of independent (14) and colluding (21) EDs are
well matched to the S.R. Then it is clear that the secrecy
outage probability linearly decreases asymptotically on the
log-log scale as the transmission power-to-noise ratio at the
UE increases for both cases. Furthermore, when the required
secrecy outage probability is 0.05, if the number of antennas
increases from 1 to 5, almost 10 dB SNR can be saved
for both cases. The above figures verified the analysis in
Section III and IV. In order to maintain clarity of presentation,
only the simulation results are shown in the following figures. Fig. 5According to the simulation results, accurate results were obtained for ϱ
close to one. V. SIMULATIONS RESULTS secrecy outage probabilities for the FD UE with different
transmission power-to-noise ratios at the UE, where dBU = 5 m, R = 50 m
and ρE = 0.005 m−2. Fig. 4. T.R. vs S.R. and approximation results (A.R.) secrecy outage
probabilities for the FD UE in the presence of different densities of EDs
where α = 2, dBU = 5 m, R = 50 m and ϱ = 1 for A.R. Fig. 4. T.R. vs S.R. and approximation results (A.R.) secrecy outage
probabilities for the FD UE in the presence of different densities of EDs,
where α = 2, dBU = 5 m, R = 50 m and ϱ = 1 for A.R. Fig. 5. T.R. vs S.R. secrecy outage probabilities for the FD UE with different
transmission power-to-noise ratios at the UE, where dBU = 5 m, R = 50 m
and ρE = 0.005 m−2. MIMO systems with a TAS-based secrecy enhancement mode)
should exploit only a small subset of independent spatial paths
to perform selection. FD scheme. Fig. 6 compares the secrecy outage probabilities
of independent (Fig. 6(a)) and colluding (Fig. 6(b)) EDs for the
HD and FD modes with respect to different λUU and α, where
dBU = 10 m, R = 50 m, K = 5 and ρE = 0.001 m−2. Hence,
in this example, we consider a more secure environment with
an average of about eight EDs located in the vicinity of the BS. It is clearly shown in the figures that as the residual self-
interference increases, the secrecy outage probability of the
FD case is adversely affected. Obviously, there is no self-
interference for the HD scheme; hence, the performance is
constant for all λUU in this figure. Of more interest is the
observation that the secrecy outage probabilities of the HD
mode are always less than for the FD mode when λUU is less
than about 11.5 dB and 10 dB for independent and colluding
cases, respectively, when PU/σ 2
U = 60 dB. Furthermore, an
important point shown in Fig. 6 is that when the path loss
exponent α increases, the enhancement of secrecy performance
by using the FD scheme will be limited due to the significant
attenuation of the jamming signal from the FD UE to the EDs. V. SIMULATIONS RESULTS It suggests that
even systems with large numbers of antennas (e.g., massive (β; dBU, R, A0) = −A0π
ϱ4R4 (4R4ϱ4d2
BU(A0 + 1/4)(ln(ϱ)2 + 2 ln(A0dBU) ln(R/ϱ) −ln(R)2)
+ R4ϱ4 ln ϱ ((A0 + 1)ϱ2 −8(A0κ −(9/4)A2
0 + (1/4)κ + 1/4)d2
BU −d4
BU A0/ϱ4)
−R4ϱ4 ln R ((A0 + 1)R2 −8(A0κ −(9/4)A2
0 + (1/4)κ + 1/4)d2
BU −d4
BU A0)
+ (R2 −ϱ2)(ϱ2(R2(A0 + 1)ϱ2 + d4
BU A0)R2 ln(A0) + ϱ2(R2(A0 + 1)ϱ2 + d4
BU A0)R2 ln(dBU)
+ R4(−A2
0 + (κ + 1)A0 + κ + 3/2)ϱ4 + A0((κ −9A0)R2 −(1/2)d2
BU A0)d4
BUϱ2 −(1/2)R2d6
BU A2
0)). (24) (β; dBU, R, A0) = −A0π
ϱ4R4 (4R4ϱ4d2
BU(A0 + 1/4)(ln(ϱ)2 + 2 ln(A0dBU) ln(R/ϱ) −ln(R)2)
+ R4ϱ4 ln ϱ ((A0 + 1)ϱ2 −8(A0κ −(9/4)A2
0 + (1/4)κ + 1/4)d2
BU −d4
BU A0/ϱ4)
−R4ϱ4 ln R ((A0 + 1)R2 −8(A0κ −(9/4)A2
0 + (1/4)κ + 1/4)d2
BU −d4
BU A0)
+ (R2 −ϱ2)(ϱ2(R2(A0 + 1)ϱ2 + d4
BU A0)R2 ln(A0) + ϱ2(R2(A0 + 1)ϱ2 + d4
BU A0)R2 ln(dBU)
+ R4(−A2
0 + (κ + 1)A0 + κ + 3/2)ϱ4 + A0((κ −9A0)R2 −(1/2)d2
BU A0)d4
BUϱ2 −(1/2)R2d6
BU A2
0)). (24) (24) CHEN et al.: SECRECY OUTAGE ANALYSIS FOR DOWNLINK TRANSMISSIONS IN THE PRESENCE OF RANDOMLY LOCATED EDs 1201 Fig. 4. T.R. vs S.R. and approximation results (A.R.) secrecy outage
probabilities for the FD UE in the presence of different densities of EDs,
where α = 2, dBU = 5 m, R = 50 m and ϱ = 1 for A.R. Fig. 5. T.R. vs S.R. secrecy outage probabilities for the FD UE with different
transmission power-to-noise ratios at the UE, where dBU = 5 m, R = 50 m
and ρE = 0.005 m−2. Fig. 4. T.R. vs S.R. and approximation results (A.R.) secrecy outage
probabilities for the FD UE in the presence of different densities of EDs,
where α = 2, dBU = 5 m, R = 50 m and ϱ = 1 for A.R. Fig. 5. T.R. vs S.R. secrecy outage probabilities for the FD UE with different
transmission power-to-noise ratios at the UE, where dBU = 5 m, R = 50 m
and ρE = 0.005 m−2. Fig. 5. T.R. vs S.R. V. SIMULATIONS RESULTS 7 shows the comparison of secrecy outage probabilities
versus different path loss exponents for the HD and FD UE
cases operating in the presence of independent and colluding
EDs, where λUU
= 0 dB, dBU
= 5 m, R = 50 m,
ρE = 0.001 m−2 and K = 1 and 5. In this example, there
are on average about eight eavesdroppers in the vicinity of the
network. We can see that the secrecy outage probability for
HD UE with independent and colluding EDs slightly decreases
until reaching a flat tail with an increasing path loss exponent. On the contrary, the secrecy outage probability for the FD According to [24], radio transmissions always encounter
a bandwidth constraint that limits maximum self-interference
cancellation. Therefore, it is useful to consider how residual
self-interference affects the secrecy outage performance of the IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 1202 Fig. 6. The comparison of secrecy outage probabilities for FD and HD UEs with different residual self-interference channel gains, where dBU = 10 m,
R = 50 m and ρE = 0.001 m−2. (a) Independent EDs. (b) Colluding EDs. Fig. 6. The comparison of secrecy outage probabilities for FD and HD UEs with different residual self-interference channel gains, where dBU = 10 m,
R = 50 m and ρE = 0.001 m−2. (a) Independent EDs. (b) Colluding EDs. Fig. 6. The comparison of secrecy outage probabilities for FD and HD UEs with different residual self-interference
R = 50 m and ρE = 0.001 m−2. (a) Independent EDs. (b) Colluding EDs. showed how secrecy performance can be enhanced by TAS and
FD communications. Our results provide useful insight and
analytical tools that can be used to develop adaptive system
solutions (examples were briefly discussed for hybrid HD/FD
UE operation) as well as a solid basis for further study. Fig. 7. The comparison of secrecy outage probabilities for FD and HD UEs
with different pathloss exponents, where ρE = 0.001 m−2, λUU = 0 dB,
dBU = 5 m and R = 50 m. APPENDIX I We assume all channels are independent and identically
distributed (i.i.d.); consequently, the cumulative distribution
function (CDF) and probability density function (PDF) of γBU
in (4) with ϖ = 0 are given by FγBU (x) =
1 −e−xdα
BU
K
=
K
k=0
Ck
K (−1)ke−kxdα
BU ,
fγBU (x) =
K
k=1
Ck
K (−1)k+1kdα
BUe−kxdα
BU ,
(25) FγBU (x) =
1 −e−xdα
BU
K
=
K
k=0
Ck
K (−1)ke−kxdα
BU ,
K fγBU (x) =
K
k=1
Ck
K (−1)k+1kdα
BUe−kxdα
BU ,
(25) (25) Fig. 7. The comparison of secrecy outage probabilities for FD and HD UEs
with different pathloss exponents, where ρE = 0.001 m−2, λUU = 0 dB,
dBU = 5 m and R = 50 m. respectively, where Ck
K = K!/[k!(K −k)!] is the binomial
coefficient. Then, the CDF of γBE∗in (6) with ϖ = 0 can be
calculated as respectively, where Ck
K = K!/[k!(K −k)!] is the binomial
coefficient. Then, the CDF of γBE∗in (6) with ϖ = 0 can be
calculated as case increases to this saturation point. The reason is that when
the UE’s transmission power fixed, the power of the jamming
signal from the FD UE is attenuated significantly for large α. Furthermore, it is clear that the secrecy outage probability for
colluding EDs is always higher than for independent EDs. FγBE∗(y) = P
max
e∈
|h B∗Ee|2
dα
BEe
< y
(a)
= E
e∈
P
|h B∗Ee|2 < ydα
BEe |
= E
e∈
1 −e−ydα
BEe
(b)
= exp
−ρE
2π
0
R
0
r
e−yrα
dr dθ
(c)
= exp
−2πρE
αy
2
α
2
α
−
2
α , yRα
(d)
= exp
−2πρE
αy
2
α
2
α
×
1 + 2πρE
αy
2
α
O(R2−α y2/α−1e−yRα)
,
(26) FγBE∗(y) = P
max
e∈
|h B∗Ee|2
dα
BEe
< y
VI. CONCLUSION In this paper, we studied a method of enhancing secrecy
performance in wireless networks with randomly located inde-
pendent and colluding EDs, which relies on the use of TAS
at the base station and an FD jamming scheme at the UE. For both of these models, we obtained expressions for the
secrecy outage probability in the downlink for HD and FD
UE operation. The expressions for HD systems have very
accurate approximate or exact forms in terms of elementary
and/or special functions for all path loss exponents. Those
related to the FD systems have very accurate approximate or
exact integral forms for general path loss exponents, while
exact closed forms are given for specific exponents. These
results have been confirmed by simulated simulations which CHEN et al.: SECRECY OUTAGE ANALYSIS FOR DOWNLINK TRANSMISSIONS IN THE PRESENCE OF RANDOMLY LOCATED EDs 1203 SNR and the ED SNR and follows the calculations presented
in Appendix I. Since the integrand is nonnegative on the
interval [0, ∞), we have the simple relations where (·) and (·, ·) denote the gamma and upper incomplete
gamma function, respectively, and where eq. (a) follows from
the independence of R.V.s {|h B∗Ee|2; Ee ∈}; eq. (b) holds
for the probability generating functional lemma [25]; eq. (c)
holds by using [26, eq. (3.326.4)]; eq. (d) follows from
the asymptotic expansion of the incomplete gamma func-
tion (R →∞) [27]. P(H)
so
> b1c1
∞
ln K
a
(1 −e−ax)K e−b1/xc1
x1+c1
dx
> b1c1
1 −1
K
K ∞
ln K
a
e−b1/xc1
x1+c1
dx
=
1 −1
K
K
1 −exp
−ac1b1
(ln K)c1
(32) According to the definition of secrecy outage probability
in (10), (25) and (26), we can obtain an approximation of the
secrecy outage probability as follows (32) P(H)
so
= 1 −
∞
0
fγBU (x)FγBE∗
x
β
dx
= 1 −
K
k=1
Ck
K (−1)k+1kdα
BU
×
∞
0
e−kxdα
BU e
−
2πρE
α
x
β
2/α ( 2
α )
dx. where the equality results from the substitution u = 1/xc1. Letting K grow large, the final line of the equation given above
becomes where the equality results from the substitution u = 1/xc1. VI. CONCLUSION Letting K grow large, the final line of the equation given above
becomes e−1
1 + O
1
K
1 −
1 −
ac1b1
(ln K)c1 + O
1
(ln K)2c1
(33) (27) and the result stated in the lemma follows. and the result stated in the lemma follows. We let We let APPENDIX III The CDF and PDF of X =
X1
X2+1 are given by FX(x) =
∞
0
Fx1(x(x2 + 1)) fx2(x2) dx2
=
K
k=0
Ck
K (−1)k
PU
dα
BU e−
kxdα
BU
PU
PU
dα
BU + kxλUU
(35) FX(x) =
∞
0
Fx1(x(x2 + 1)) fx2(x2) dx2 =
K
k=0
Ck
K (−1)k
PU
dα
BU e−
kxdα
BU
PU
PU
dα
BU + kxλUU
(35) (35) ×
n=0
s + 1 + n
2q
ds
=
√pq
a2
p+2q−3
2
π
p+2q
2
−1
× G p+2q,0
0,p+2q
a2q
k b p
p p4qq2q
−
0, 1
p, ..., p−1
p , 1
2q , 2
2q , ..., 1
,
(30)
and
fX(x)
=
K
k=1
Ck
K (−1)k+1 (PU + kxλUUdα
BU + PUλUU)ke−
kxdα
BU
PU
dα
BU( PU
dα
BU + kxλUU)2
. (36)
Th
l tti
Y
|hB∗Ee|2
/ |hU Ee |2
it i
ibl
t (36)
(30) Then letting Y = max
e∈
|hB∗Ee|2
dα
BEe
/ |hU Ee |2
dα
U Ee
, it is possible to
show that the CDF of Y can be written as (37), shown at the
top of the next page,
where where G(·) denotes Meijer’s G furcation, u > 0 and (a) holds
from the multiplication theorem [27]. where G(·) denotes Meijer’s G furcation, u > 0 and (a) holds
from the multiplication theorem [27]. APPENDIX III I =
∞
0
e−axe−b
xc dx =
∞
0
ue−aue−
b1/c
x
c du
u
(28) (28) According to (10), (4) and (6) with ϖ
= 1, we let
X1 = PU
max
k∈(1...K)(|h BkU|2) and X2 = |hUU|2. Then after
self-interference cancellation, the average channel gain of the
residual self-interference can be denoted as λUU. Therefore,
the CDF of X1 and the PDF of X2 can be written as According to (10), (4) and (6) with ϖ
= 1, we let
X1 = PU
max
k∈(1...K)(|h BkU|2) and X2 = |hUU|2. Then after where a = kdα
BU, b =
2πρE
α
( 2q
p )β2q/p and c = 2q/p. By using the Mellin convolution theorem, we can get the
Mellin transform as where a = kdα
BU, b =
2πρE
α
( 2q
p )β2q/p and c = 2q/p. By using the Mellin convolution theorem, we can get the
Mellin transform as (
)
self-interference cancellation, the average channel gain of the
residual self-interference can be denoted as λUU. Therefore,
the CDF of X1 and the PDF of X2 can be written as M[I; s] =
p
2qas+1
ps
2q
(1 + s). APPENDIX III (29) (29) FX1(x1) =
K
k=0
Ck
K (−1)ke−
kx1dα
BU
PU
fX2(x2) = 1/λUUe−x2/λUU , FX1(x1) =
K
k=0
Ck
K (−1)ke−
kx1dα
BU
PU
fX2(x2) = 1/λUUe−x2/λUU ,
(34) Then the inverse transform can be written as fX2(x2) = 1/λUUe−x2/λUU ,
(34) (34) I =
p
2πia
u+i∞
u−i∞
(ps)
2q(s + 1
2q )
(a2qb p)−s ds
(a)
=
√pq
a2
p+2q−3
2
π
p+2q
2
−1
1
2πi
×
u+i∞
u−i∞
a2qb p
p p4qq2q
−s p−1
n=0
s + n
p
×
2q−1
n=0
s + 1 + n
2q
ds
=
√pq
a2
p+2q−3
2
π
p+2q
2
−1
× G p+2q,0
0,p+2q
a2q
k b p
p p4qq2q
−
0, 1
p, ..., p−1
p , 1
2q , 2
2q , ..., 1
,
(30 I =
p
2πia
u+i∞
u−i∞
(ps)
2q(s + 1
2q )
(a2qb p)−s ds
(a)
=
√pq
a2
p+2q−3
2
π
p+2q
2
−1
1
2πi
×
u+i∞
u−i∞
a2qb p
p p4qq2q
−s p−1
n=0
s + n
p
×
2q−1
n=0
s + 1 + n
2q
ds
=
√pq
a2
p+2q−3
2
π
p+2q
2
−1
× G p+2q,0
0,p+2q
a2q
k b p
p p4qq2q
−
0, 1
p, ..., p−1
p , 1
2q , 2
2q , ..., 1
,
(30)
fX2(x2) = 1/λUUe−x2/λUU ,
respectively. The CDF and PDF of X =
X1
X2+1 are gi
FX(x) =
∞
0
Fx1(x(x2 + 1)) fx2(x2) dx2
=
K
k=0
Ck
K (−1)k
PU
dα
BU e−
kxdα
BU
PU
PU
dα
BU + kxλUU
and
fX(x)
=
K
k=1
Ck
K (−1)k+1 (PU + kxλUUdα
BU + PUλUU)ke
dα
BU( PU
dα
BU + kxλUU)2
Then letting Y
max
|hB∗Ee|2
/ |hU Ee |2
it is po respectively. The CDF and PDF of X =
X1
X2+1 are given by respectively. IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 Then by using (36) and (37), the secrecy outage
probability of the FD UE can be written as E
e−sZ
|s=kβdα
BU
= E
e∈
e−kβdα
BU |hB∗Ee |2d−α
BEe
= E
e∈
E|hB∗Ee|2
e−kβdα
BU |hB∗Ee|2d−α
BEe
(a)
= E
e∈
∞
0
e−kβdα
BU td−α
BEee−t dt
= E
e∈
1
1 + kβ(dBU/dBEe)α
(b)
= exp
−ρE
2π
0
R
0
1 −
1
1 + kβ(dBU/r)α
r dr dθ
= exp
−π R2ρE F
1, 2
α ; 1 + 2
α ; −
Rα
kβdα
BU
,
(42) E
e−sZ
|s=kβdα
BU
= E
e∈
e−kβdα
BU |hB∗Ee |2d−α
BEe
= E
e∈
E|hB∗Ee|2
e−kβdα
BU |hB∗Ee|2d−α
BEe
(a)
= E
e∈
∞
0
e−kβdα
BU td−α
BEee−t dt
= E
e∈
1
1 + kβ(dBU/dBEe)α
(b)
= exp
−ρE
2π
0
R
0
1 −
1
1 + kβ(dBU/r)α
r dr dθ
= exp
−π R2ρE F
1, 2
α ; 1 + 2
α ; −
Rα
kβdα
BU
,
(42) P(F)
so
≤1 −
∞
0
fX(x)FY
x
β
dx,
(40) (40) which has been shown in Proposition 3. which has been shown in Proposition 3. IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 1204 FY (y) = P
⎛
⎜⎝max
e∈
⎛
⎜⎝
|hB∗Ee|2
dα
BEe
|hU Ee |2
dα
U Ee
⎞
⎟⎠< y
⎞
⎟⎠= E
P
max
e∈
|h B∗Ee|2d−α
BEe
|hU Ee|2d−α
U Ee
< y |
(a)
= E
e∈
P
|h B∗Ee|2
|hU Ee|2 < y
dα
BEe
dα
U Ee
|
(b)
= E
e∈
ydα
BEe
ydα
BEe + dα
U Ee
(c)
= exp
−ρE
R
0
2π
0
r(y;r, θ) dθ dr
. (37) (37) E
e−sZ
|s= 2kβ
PU dα
BU = E
⎡
⎢⎣
e∈
e
−2kβ
PU dα
BU
td−α
BEe
d2
BEe +d2
BU −2dBEe dBU cos(θ)
−α
⎤
⎥⎦
= E
⎡
⎢⎣
e∈
Et
⎡
⎢⎣e
−2kβ
PU dα
BU
td−α
BEe
d2
BEe +d2
BU −2dBEe dBU cos(θ)
−α
⎤
⎥⎦
⎤
⎥⎦
(a)
= E
⎡
⎢⎣
e∈
∞
0
e
−2kβ
PU dα
BU t
dBEe
d2
BEe +d2
BU −2dBEe dBU cos(θ)
−α
1
(1 + t)2 dt
⎤
⎥⎦
(b)
= exp
−ρE
R
0
2π
0
AeAE1 (A)r dθ dr
(c)
≃exp
−ρE
ϱ
0
2π
0
(1 −1/A)r dr dθ +
R
ϱ
2π
0
A(A + 1) (A −ln(A) −κ)r dr dθ
(d)
= exp
⎛
⎜⎝−ρE
⎛
⎜⎝πϱ2 −π PU
2kβ
⎛
⎜⎝ln
⎛
⎜⎝
1
1 −
ϱ
dBU
2
⎞
⎟⎠+
ϱ
dBU
2
⎞
⎟⎠+ (β; dBU, R, A0)
⎞
⎟⎠
⎞
⎟⎠
. APPENDIX II (y;r, θ) = 1 −
yrα
yrα + (
r2 + d2
BU −2rdBUcosθ)α
,
(38) (38) We begin with the following basic integral definition of the
secrecy outage probability for this case P(H)
so
= b1c1
∞
0
(1 −e−ax)K e−b1/xc1
x1+c1
dx
(31) and (a) follows from the independence of |hB∗Ee|2
|hU Ee |2 ; Ee ∈, (b)
holds since the CDF (31) Fν(ν) = P
|h B∗Ee|2
|hU Ee|2 < ν
=
ν
ν + dα
U E/dα
BE
,
(39) where a = βdα
BU, b1 = c1πρE (2/α) and c1 = 2/α. This
expression can easily be derived from the definitions of the UE (39) 1204
IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017
FY (y) = P
⎛
⎜⎝max
e∈
⎛
⎜⎝
|hB∗Ee|2
dα
BEe
|hU Ee |2
dα
U Ee
⎞
⎟⎠< y
⎞
⎟⎠= E
P
max
e∈
|h B∗Ee|2d−α
BEe
|hU Ee|2d−α
U Ee
< y |
(a)
= E
e∈
P
|h B∗Ee|2
|hU Ee|2 < y
dα
BEe
dα
U Ee
|
(b)
= E
e∈
ydα
BEe
ydα
BEe + dα
U Ee
(c)
= exp
−ρE
R
0
2π
0
r(y;r, θ) dθ dr
. (37)
E
e−sZ
|s= 2kβ
PU dα
BU = E
⎡
⎢⎣
e∈
e
−2kβ
PU dα
BU
td−α
BEe
d2
BEe +d2
BU −2dBEe dBU cos(θ)
−α
⎤
⎥⎦
= E
⎡
⎢⎣
e∈
Et
⎡
⎢⎣e
−2kβ
PU dα
BU
td−α
BEe
d2
BEe +d2
BU −2dBEe dBU cos(θ)
−α
⎤
⎥⎦
⎤
⎥⎦
(a)
= E
⎡
⎢⎣
e∈
∞
0
e
−2kβ
PU dα
BU t
dBEe
d2
BEe +d2
BU −2dBEe dBU cos(θ)
−α
1
(1 + t)2 dt
⎤
⎥⎦
(b)
= exp
−ρE
R
0
2π
0
AeAE1 (A)r dθ dr
(c)
≃exp
−ρE
ϱ
0
2π
0
(1 −1/A)r dr dθ +
R
ϱ
2π
0
A(A + 1) (A −ln(A) −κ)r dr dθ
(d)
= exp
⎛
⎜⎝−ρE
⎛
⎜⎝πϱ2 −π PU
2kβ
⎛
⎜⎝ln
⎛
⎜⎝
1
1 −
ϱ
dBU
2
⎞
⎟⎠+
ϱ
dBU
2
⎞
⎟⎠+ (β; dBU, R, A0)
⎞
⎟⎠
⎞
⎟⎠
. IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 (44) E
e−sZ
|s= 2kβ
PU dα
BU = E
⎡
⎢⎣
e∈
e
−2kβ
PU dα
BU
td−α
BEe
d2
BEe +d2
BU −2dBEe dBU cos(θ)
−α
⎤
⎥⎦
= E
⎡
⎢⎣
e∈
Et
⎡
⎢⎣e
−2kβ
PU dα
BU
td−α
BEe
d2
BEe +d2
BU −2dBEe dBU cos(θ)
−α
⎤
⎥⎦
⎤
⎥⎦
(a)
= E
⎡
⎢⎣
e∈
∞
0
e
−2kβ
PU dα
BU t
dBEe
d2
BEe +d2
BU −2dBEe dBU cos(θ)
−α
1
(1 + t)2 dt
⎤
⎥⎦
(b)
= exp
−ρE
R
0
2π
0
AeAE1 (A)r dθ dr
(c)
≃exp
−ρE
ϱ
0
2π
0
(1 −1/A)r dr dθ +
R
ϱ
2π
0
A(A + 1) (A −ln(A) −κ)r dr dθ
(d)
= exp
⎛
⎜⎝−ρE
⎛
⎜⎝πϱ2 −π PU
2kβ
⎛
⎜⎝ln
⎛
⎜⎝
1
1 −
ϱ
dBU
2
⎞
⎟⎠+
ϱ
dBU
2
⎞
⎟⎠+ (β; dBU, R, A0)
⎞
⎟⎠
⎞
⎟⎠
. (44) (44) where Z = ! e∈
|hB∗Ee|2
dα
BEe
and E "e−sZ# |s=kβdα
BU is given by where Z = ! e∈
|hB∗Ee|2
dα
BEe
and E "e−sZ# |s=kβdα
BU is given by
E
e−sZ
|s=kβdα
BU
= E
e∈
e−kβdα
BU |hB∗Ee |2d−α
BEe
= E
e∈
E|hB∗Ee|2
e−kβdα
BU |hB∗Ee|2d−α
BEe
(a)
= E
e∈
∞
0
e−kβdα
BU td−α
BEee−t dt
= E
e∈
1
1 + kβ(dBU/dBEe)α
(b)
= exp
−ρE
2π
0
R
0
1 −
1
1 + kβ(dBU/r)α
r dr dθ
= exp
−π R2ρE F
1, 2
α ; 1 + 2
α ; −
Rα
kβdα
BU
,
(42)
where, for brevity and ease of exposition, we let t = |h B∗Ee|2
in (a) and the PDF of t is e−t, F(a, b; c; z) denotes the
Gaussian hypergeometric function, and (b) holds for the prob-
ability generating functional lemma [25]. and (c) holds for the probability generating functional
lemma [25]. APPENDIX II (44)
and (c) holds for the probability generating functional
lemma [25]. Then by using (36) and (37), the secrecy outage
probability of the FD UE can be written as
where Z = ! e∈
|hB∗Ee|2
dα
BEe
and E "e−sZ# |s=kβdα
BU is given by
E
e−sZ
| APPENDIX V [6] H. Weingarten, T. Liu, S. Shamai (Shitz), Y. Steinberg, and P. Viswanath,
“The capacity region of the degraded multiple-input multiple-output
compound broadcast channel,” IEEE Trans. Inf. Theory, vol. 55, no. 11,
pp. 5011–5023, Nov. 2009. According to the definition of secrecy outage probability
in (10), (4) and (6) with ϖ = 1, modeling the residual
self-interference as AWGN noise [28], [29] and ignoring the
noise at ED as in [21]–[23], we can obtain the secrecy outage
probability as follows [7] G. Chen, Y. Gong, P. Xiao, and J. A. Chambers, “Physical layer network
security in the full-duplex relay system,” IEEE Trans. Inf. Forensics
Security, vol. 10, no. 3, pp. 574–583, Mar. 2015. [8] G. Chen, Y. Gong, P. Xiao, and J. A. Chambers, “Dual antenna selection
in secure cognitive radio networks,” IEEE Trans. Veh. Technol., vol. 65,
no. 10, pp. 7993–8002, Oct. 2016. P(F)
so
≤P
⎛
⎜⎜⎜⎜⎜⎜⎜⎝
PB
2
max
k∈(1...K)
|hBkU |2
dα
BU
! e∈
⎛
⎝
PB
|hB∗Ee |2
dα
BEe
PU
|hU Ee |2
dα
U Ee
⎞
⎠
< β
⎞
⎟⎟⎟⎟⎟⎟⎟⎠
= P
⎛
⎜⎝max
k∈(1...K)
|h BkU|2
dα
BU
< 2β
PU
e∈
⎛
⎜⎝
|hB∗Ee|2
dα
BEe
|hU Ee |2
dα
U Ee
⎞
⎟⎠
⎞
⎟⎠
= 1 +
K
k=1
Ck
K (−1)k
∞
0
e−2kβ
PU zdα
BU fZ(z)dz
= 1 +
K
k=1
Ck
K (−1)kE
e−sZ
|s= 2kβ
PU dα
BU ,
(43)
where Z = ! e∈
⎛
⎝
|hB∗Ee |2
dα
BEe
|hU Ee |2
dα
U Ee
⎞
⎠and E "e−sZ# |s= 2kβ
PU dα
BU can be [9] L. Dong, Z. Han, A. P. Petropulu, and H. V. Poor, “Amplify-and-forward
based cooperation for secure wireless communications,” in Proc. IEEE
Int. Conf. Acoust., Speech, Signal Process., Taipei, Taiwan, Apr. 2009,
pp. 2613–2616. [10] M. Haenggi, “The secrecy graph and some of its properties,” in Proc. IEEE Int. Symp. Inf. Theory, Toronto, Canada, Jul. 2008, pp. 539–543. [11] P. C. Pinto, J. Barros, and M. Z. Win, “Physical-layer security in
stochastic wireless networks,” in Proc. 11th IEEE Singapore Int. Conf. Cuommun. Syst., Singapore, Nov. 2008, pp. 974–979. [12] X. Zhou, R. K. Ganti, J. G. Andrews, and A. Hjørungnes, “On
the throughput cost of physical layer security in decentralized wire-
less networks,” IEEE Trans. Wireless Commun., vol. 10, no. 8,
pp. 2764–2775, Aug. 2011. pp
g
[13] G. APPENDIX V Geraci, S. Singh, J. G. Andrews, J. Yuan, and I. B. Collings,
“Secrecy rates in broadcast channels with confidential messages and
external eavesdroppers,” IEEE Trans. Wireless Commun., vol. 13, no. 5,
pp. 2931–2943, May 2014. [14] T. X. Zheng, H. M. Wang, and Q. Yin, “On transmission secrecy outage
of a multi-antenna system with randomly located eavesdroppers,” IEEE
Commun. Lett., vol. 18, no. 8, pp. 1299–1302, Aug. 2014. pp
g
[15] M. Ghogho and A. Swami, “Physical-layer secrecy of MIMO commu-
nications in the presence of a poisson random field of eavesdroppers,”
in Proc. IEEE ICC, Jun. 2011, pp. 1–5. pp
[16] T. X. Zheng, H. M. Wang, J. Yuan, D. Towsley, and M. H. Lee, “Multi-
antenna transmission with artificial noise against randomly distributed
eavesdroppers,” IEEE Trans. Commun., vol. 63, no. 11, pp. 4347–4362,
Nov. 2015. ⎝
U Ee
⎠
obtained as (44) shown at the top of the previous page. ⎝
U Ee
⎠
obtained as (44) shown at the top of the previous page. For
brevity
and
ease
of
exposition,
we
let
t
=
|hB∗Ee|2
|hU E |2
in
(a)
and
the
PDF
of
t
is
1/(1 + t)2, [17] G. Chen, Y. Gong, and J. Chambers, “Study of relay selection in a multi-
cell cognitive network,” IEEE Wireless Commun. Lett., vol. 2, no. 4,
pp. 435–438, Aug. 2013. |hU Ee |
A
=
2kβ
PU dα
BU
r
r2+d2
BU −2rdBU cos(θ)
−α
, A0
=
2kβ
PU d2
BU, [18] P. C. Pinto, J. Barros, and M. Z. Win, “Secure communication in sto-
chastic wireless networks—part II: Maximum rate and collusion,” IEEE
Trans. Inf. Forensics Security, vol. 7, no. 1, pp. 139–147, Feb. 2012. [19] X. Zhou, R. K. Ganti, and J. G. Andrews, “Secure wireless network
connectivity with multi-antenna transmission,” IEEE Trans. Wireless
Commun., vol. 10, no. 2, pp. 425–430, Feb. 2011. (β; dBU, R, A0) is given as (24) and (b) holds for the proba-
bility generating functional lemma [25]. In (c), the first double
integral can be approximately obtained by using asymptotic
(divergent) series [30] and the second double integral can be
approximated by using the Taylor series [31], and (d) holds
when α = 2. [20] P. C. Pinto, J. Barros, and M. Z. Win, “Secure communication in
stochastic wireless networks—Part I: Connectivity,” IEEE Trans. Inf. Forensics Security, vol. 7, pp. 125–138, Feb. 2012. y
pp
[21] X. Zhang, X. APPENDIX V Zhou, and M. R. McKay, “Enhancing secrecy with multi-
antenna transmission in wireless ad hoc networks,” IEEE Trans. Inf. Forensics Security, vol. 8, no. 11, pp. 1802–1814, Nov. 2013. [22] C. Wang, H. M. Wang, X. G. Xia, and C. Liu, “Uncoordinated
jammer selection for securing SIMOME wiretap channels: A stochastic
geometry approach,” IEEE Trans. Wireless Commun., vol. 14, no. 5,
pp. 2596–2612, May 2015. ACKNOWLEDGEMENTS The authors wish to thank Prof. C. Dettmann, Dr. K. Koufos
and Dr. D. Simmons for their input. They also would like
to thank the anonymous reviewers and the editor for their
constructive comments. [23] X. Zhang, X. Zhou, and M. R. McKay, “On the design of artificial-
noise-aided secure multi-antenna transmission in slow fading channels,”
IEEE Trans. Veh. Technol., vol. 62, no. 5, pp. 2170–2181, Jun. 2013. [24] S. Hong et al., “Applications of self-interference cancellation in 5G and
beyond,” IEEE Commun. Mag., vol. 52, no. 2, pp. 114–121, Feb. 2014. CHEN et al.: SECRECY OUTAGE ANALYSIS FOR DOWNLINK TRANSMISSIONS IN THE PRESENCE OF RANDOMLY LOCATED EDs 1205 APPENDIX IV According to the definition of secrecy outage probabil-
ity (10), (4) and (6) with ϖ = 0, we can obtain the secrecy
outage probability as followed P(H)
so
= P
⎛
⎜⎜⎝
max
k∈(1...K)
|hBkU |2
dα
BU
! e∈
|hB∗Ee |2
dα
BEe
< β
⎞
⎟⎟⎠
= P
max
k∈(1...K)
|h BkU|2
dα
BU
< β
e∈
|h B∗Ee|2
dα
BEe
=
K
k=0
Ck
K (−1)k
∞
0
e−kβzdα
BU fZ(z) dz
=
K
k=0
Ck
K (−1)kE
e−sZ
|s=kβdα
BU
(4 P(H)
so
= P
⎛
⎜⎜⎝
max
k∈(1...K)
|hBkU |2
dα
BU
! e∈
|hB∗Ee |2
dα
BEe
< β
⎞
⎟⎟⎠
= P
max
k∈(1...K)
|h BkU|2
dα
BU
< β
e∈
|h B∗Ee|2
dα
BEe
=
K
k=0
Ck
K (−1)k
∞
0
e−kβzdα
BU fZ(z) dz
=
K
k=0
Ck
K (−1)kE
e−sZ
|s=kβdα
BU
(41 (42) where, for brevity and ease of exposition, we let t = |h B∗Ee|2
in (a) and the PDF of t is e−t, F(a, b; c; z) denotes the
Gaussian hypergeometric function, and (b) holds for the prob-
ability generating functional lemma [25]. (41) IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 1206 Gaojie Chen (S’09–M’12) received the B. Eng. and B. Ec. degree in electrical information engi-
neering and international economics and trade from
the Northwest University, Xi’an, China, in 2006,
and the M.Sc. (Distinction) and Ph.D. degrees in
electrical and electronic engineering from Lough-
borough University, Loughborough, U.K., in 2008
and 2012, respectively. From 2008 to 2009, he was
a Software Engineering with DTmobile, Beijing,
China, and from 2012 to 2013 a Research Associate
with the School of Electronic, Electrical and Sys-
tems Engineering, Loughborough University, Loughborough, U.K. He was
a Research Fellow with the 5GIC, the Faculty of Engineering and Physical
Sciences, University of Surrey, U.K., from 2014 to 2015. He is currently a
Research Associate with the Department of Engineering Science, University of
Oxford, U.K. His current research interests include information theory, wire-
less communications, cooperative communications, cognitive radio, secrecy
communication, and random geometric networks. Marco
Di
Renzo
(S’05–AM’07–M’09–SM’14)
received the Laurea degree (cum laude) and the
Ph.D. degree in electrical engineering from the Uni-
versity of L’Aquila, Italy, in 2003 and 2007, respec-
tively, and the D.Sc. degree (Habilitation diriger
des recherches) from the University of Paris-Sud,
France, in 2013. Since 2010, he has been a CNRS
Associate Professor (Charg de Recherche Titulaire
CNRS) with the Laboratory of Signals and Sys-
tems, Paris-Saclay University-CNRS, CentraleSu-
plec, University of Paris-Sud, Paris, France. He is
the Project Coordinator of two EU-funded multi-partner projects (ETN-
5Gwireless and ETN-5Gaura). His research interests include wireless com-
munications, communication theory, and stochastic geometry. He currently
serves as an Editor of the IEEE COMMUNICATIONS LETTERS and IEEE
TRANSACTIONS ON COMMUNICATIONS. He is currently a Distinguished
Lecturer of Communications Society and the IEEE Vehicular Technology
Society. He was a recipient of several research distinctions, which include
the 2013 Network of Excellence NEWCOM Best Paper Award, the 2013
IEEE-COMSOC Best Young Researcher Award for Europe, Middle East and
Africa (EMEA Region), the 2015 IEEE Jack Neubauer Memorial Best System
Paper Award, the 2015 Distinguished Visiting Fellow of the Royal Academy
of Engineering, U.K., the 2015–2018 CNRS Award for Excellence in Research
and in Advising Doctoral Students, the 2016 MSCA Global Fellowship, and
six Best Paper Awards at IEEE conferences. Justin P. Coon (S’02–M’05–SM’10) received the
B.Sc. degree (Hons.) in electrical engineering from
the Calhoun Honours College, Clemson Univer-
sity, USA, and the Ph.D. REFERENCES [25] M. Haenggi, Stochastic Geometry for Wireless Networks. Cambridge,
U.K.: Cambridge Univ. Press, 2012. [1] A. D. Wyner, “The wire-tap channel,” Bell Syst. Tech. J., vol. 54, no. 8,
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Products. Amsterdam, The Netherlands: Elsevier, 2007. [2] E. Tekin and A. Yener, “The general Gaussian multiple access and
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IEEE Trans. Inf. Theory, vol. 54, no. 6, pp. 2735–2751, Jun. 2008. [27] M. Abramowitz and I. A. Stegun, Handbook of Mathematical Functions
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[3] P. Popovski and O. Simeone, “Wireless secrecy in cellular systems with
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Scientists and Engineers. New York, NY, USA: McGraw-Hill, 1978. IEEE TRANSACTIONS ON INFORMATION FORENSICS AND SECURITY, VOL. 12, NO. 5, MAY 2017 degree in communica-
tions from the University of Bristol, U.K., in 2000
and 2005, respectively. In 2004, he joined Toshiba
Research Europe Ltd., as a Research Engineer with
Bristol-based Telecommunications Research Labora-
tory (TRL), where he was involved on a broad range
of communication technologies and theories, includ-
ing single and multicarrier modulation techniques,
estimation and detection, diversity methods, system performance analysis, and
networks. He held the position of Research Manager from 2010 to 2013, where
he led all theoretical and applied research on the physical layer at TRL. He
was a Visiting Fellow with the School of Mathematics, University of Bristol,
from 2010 to 2012, where he held a Reader position with the Department
of Electrical and Electronic Engineering from 2012 to 2013. He joined the
University of Oxford in 2013, where he is currently an Associate Professor
with the Department of Engineering Science and a Tutorial Fellow of the
Oriel College. He has authored over 100 papers in leading international journals and
conferences, and is a named inventor on over 30 patents. He is the Technical
Manager of the EU FP7 project DIWINE. His research interests include
communication theory, information theory, and network theory. He was a
recipient of TRL’s Distinguished Research Award for block-spread CDMA,
aspects of which have been adopted as mandatory features in the 3GPP
LTE Rel-8 standard. He was also a co-recipient of two best paper awards
presented at the ISWCS 13 and the EuCNC 14. He has served as an Editor
of IEEE TRANSACTIONS ON WIRELESS COMMUNICATIONS from 2007 to
2013, IEEE TRANSACTIONS ON VEHICULAR TECHNOLOGY from 2013 to
2016 (received the award for Outstanding Contribution in 2014), and IEEE
WIRELESS COMMUNICATIONS LETTERS in 2016.
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Mitochondria Regulate the Differentiation of Stem Cells from Human Exfoliated Deciduous Teeth
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Hiromichi Kato
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English
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Spoken
| 8,149
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Mitochondria Regulate the Differentiation of Stem Cells from Human Exfoliated
Deciduous Teeth Hiroki Kato1*, Thanh Thi Mai Pham1, Haruyoshi Yamaza1, Keiji Masuda1, Yuta Hirofuji1, Xu Han1,
Hiroshi Sato1, Tomoaki Taguchi2, and Kazuaki Nonaka1** Hiroki Kato1*, Thanh Thi Mai Pham1, Haruyoshi Yamaza1, Keiji Masuda1, Yuta Hirofuji1, Xu
Hiroshi Sato1, Tomoaki Taguchi2, and Kazuaki Nonaka1** 1Section of Oral Medicine for Child, Division of Oral Health, Growth & Development, Faculty of Dental
Science, Kyushu University, Maidashi 3-1-1, Higashi-Ku, Fukuoka 812-8582, Japan, 2Department of
Pediatric Surgery, Reproductive and Developmental Medicine, Graduate School of Medical Sciences,
Kyushu University, Maidashi 3-1-1, Higashi-Ku, Fukuoka 812-8582, Japan ABSTRACT. Stem cells from human exfoliated deciduous teeth (SHED) are isolated from the dental pulp tissue
of primary teeth and can differentiate into neuronal cells. Although SHED are a desirable type of stem cells for
transplantation therapy and for the study of neurological diseases, a large part of the neuronal differentiation
machinery of SHED remains unclear. Recent studies have suggested that mitochondrial activity is involved in the
differentiation of stem cells. In the present work, we investigated the neuronal differentiation machinery of SHED
by focusing on mitochondrial activity. During neuronal differentiation of SHED, we observed increased
mitochondrial membrane potential, increased mitochondrial DNA, and elongated mitochondria. Furthermore, to
examine the demand for mitochondrial activity in neuronal differentiation, we then differentiated SHED into
neuronal cells in the presence of rotenone, an inhibitor of mitochondrial respiratory chain complex I, and
carbonyl cyanide m-chlorophenyl hydrazone (CCCP), a mitochondrial uncoupler, and found that neuronal
differentiation was inhibited by treatment with rotenone and CCCP. These results indicated that increased
mitochondrial activity was crucial for the neuronal differentiation of SHED. Key words:
mitochondria, differentiation, stem cells, dental pulp, exfoliated deciduous teeth Copyright: ©2017 The Author(s). This is an open access article distributed under the terms of the Creative Commons
BY (Attribution) License (https://creativecommons.org/licenses/by/4.0/legalcode), which permits the unrestricted
distribution, reproduction and use of the article provided the original source and authors are credited. Copyright: ©2017 The Author(s). This is an open access article distributed under the terms of the Creative Commons
BY (Attribution) License (https://creativecommons.org/licenses/by/4.0/legalcode), which permits the unrestricted
distribution, reproduction and use of the article provided the original source and authors are credited. Copyright: ©2017 The Author(s). This is an open access article distributed under the terms of the Creative Commons
BY (Attribution) License (https://creativecommons.org/licenses/by/4.0/legalcode), which permits the unrestricted
distribution, reproduction and use of the article provided the original source and authors are credited. Copyright: ©2017 The Author(s). This is an open access article distributed under the terms of the Creative Commons
BY (Attribution) License (https://creativecommons.org/licenses/by/4.0/legalcode), which permits the unrestricted
distribution, reproduction and use of the article provided the original source and authors are credited. CELL STRUCTURE AND FUNCTION 42: 105–116 (2017) *To whom correspondence should be addressed: Hiroki Kato, Section of
Oral Medicine for Child, Division of Oral Health, Growth & Develop-
ment, Faculty of Dental Science, Kyushu University, Maidashi 3-1-1,
Higashi-Ku, Fukuoka 812-8582, Japan. *To whom correspondence should be addressed: Hiroki Kato, Section of
Oral Medicine for Child, Division of Oral Health, Growth & Develop-
ment, Faculty of Dental Science, Kyushu University, Maidashi 3-1-1,
Higashi-Ku, Fukuoka 812-8582, Japan.
Tel: +81–92–642–6402, Fax: +81–92–642–6488
E-mail: kato@dent.kyushu-u.ac.jp
**To whom correspondence should be addressed: Kazuaki Nonaka, Sec-
tion of Oral Medicine for Child, Division of Oral Health, Growth &
Development, Faculty of Dental Science, Kyushu University, Maidashi
3-1-1, Higashi-Ku, Fukuoka 812-8582, Japan.
Tel: +81–92–642–6402, Fax: +81–92–642–6488
E-mail: nonaka@dent.kyushu-u.ac.jp Isolation and culture of SHED An exfoliated deciduous tooth was collected from a 4-year-old
child at Pediatric & Special Needs Dentistry, Kyushu University
Hospital, with the permission of the child’s parents. The isolation
and culture of SHED were performed as described previously
(Miura et al., 2003) with small modifications. The dental pulp was
gently isolated from a residual crown and treated with 3 mg/mL
collagenase I (Washington, NJ, USA) and 4 mg/mL dispase II
(Wako Pure Chemical Industries, Osaka, Japan) in phosphate-
buffered saline (PBS) containing 2 mM CaCl2 for 60 min at 37°C. Next, the undigested tissues and cell aggregates were removed by
filtration through a 70-μm cell strainer (Corning, NY, USA). Single-cell suspensions were then seeded into a 6-well plate
(Corning) and cultured in alpha modification of Eagle’s medium
(α-MEM; Sigma-Aldrich, MO, USA) supplemented with 15%
fetal bovine serum (FBS; Sigma-Aldrich), 100 μM L-ascorbic acid
2-phosphate (Wako Pure Chemical Industries), 2 mM L-glutamine
(Life Technologies, NY, USA), 250 μg/mL fungizone (Life Tech-
nologies), 100 U/mL penicillin, and 100 μg/mL streptomycin
(Life Technologies) at 37°C in an atmosphere containing 5% CO2. SHED at passages 5–10 were used in this study. For sphere forma-
tion, 2×105 SHED were cultured in 6-well plates for suspension Neural differentiation of SHED The differentiation of SHED into neuronal cells was performed as
described previously (Miura et al., 2003). To differentiate SHED
into neuronal cells, SHED were cultured in neurobasal A medium
(Life Technologies) supplemented with 2% B27 supplement (Life
Technologies), 40 ng/mL basic fibroblast growth factor (bFGF;
Peprotech, NJ, USA), and 20 ng/mL epidermal growth factor
(EGF; Sigma-Aldrich) for 2 or 10 days at 37°C in an atmosphere
containing 5% CO2. The medium was changed after 5 days. In this study, to examine the demand for mitochondrial
activity in the neuronal differentiation of SHED, we ana-
lyzed the mitochondrial membrane potential (MMP), mito-
chondrial mass, mtDNA contents, and mitochondrial
morphology during neuronal differentiation. We also exam-
ined the effects of rotenone, a mitochondrial inhibitor, and
carbonyl cyanide 3-chlorophenylhydrazone (CCCP), a
mitochondrial uncoupler, on the neuronal differentiation of
SHED. Our findings will provide a platform for further
studies focusing on the use of SHED in clinical research
and drug discovery focusing on mitochondrial activity. Immunocytochemistry Cells cultured on coverslips were fixed with 4% paraformalde-
hyde in 0.1 M sodium phosphate buffer (pH 7.4) for 10 min at
room temperature and then permeabilized with 0.1% Triton X-100
in PBS for 5 min. For the detection of N-methyl-D-aspartate
receptor subunit 1 (NMDAR1), the permeabilization step was
skipped. The cells were blocked with 2% bovine serum albumin
(BSA; Wako Pure Chemical Industries) in PBS for 20 min and
then incubated with the following primary antibodies: anti-
STRO-1 (Millipore, CA, USA), anti-Nestin (Millipore), anti-β-
tubulin III (Sigma-Aldrich), anti-microtubule-associated protein
2 (MAP2; Sigma-Aldrich), anti-Tom20 (Santa Cruz Biotechnol-
ogy, CA, USA), and anti-NMDAR1 (Millipore). After 90 min, the
cells were incubated with Alexa Fluor-conjugated secondary anti-
bodies (Life Technologies) in the dark for 60 min. After staining
with antibodies, the nuclei were stained with 1 μg/mL 4',6-
diamidino-2-phenylindole
dihydrochloride
(DAPI;
Dojindo,
Kumamoto, Japan). The cells were then mounted using ProLong
Diamond (Life Technologies). Fluorescent images were captured
using an Axio Imager M2 (Zeiss, Oberkochen, Germany) equip-
ped with ApoTome2 (Zeiss). The length of the cells was measured
and analyzed using MetaMorph (Molecular Devices, CA, USA). To evaluate the cell numbers for 10 days after neuronal differen-
tiation, 5 images of the cells stained with anti-β-tubulin III and
DAPI were randomly taken from each of the 5 experiments. The
cell numbers were then analyzed using MetaMorph (Molecular
Devices). Ethics statement Experiments using human samples were reviewed and approved
by the Kyushu University Institutional Review Board for Human
Genome/Gene Research and were conducted in accordance with
the Declaration of Helsinki. Introduction ation capacity; SHED also show multidifferentiation poten-
tial and can differentiate into neuronal, osteogenic,
adipogenic, and hepatogenic cells (Ma et al., 2012; Miura
et al., 2003; Yamaza et al., 2015). SHED have been shown
to be able to differentiate into neurons, astrocytes, and oli-
godendrocytes (Jarmalavičiūtė et al., 2013; Miura et al.,
2003; Nourbakhsh et al., 2011; Sakai et al., 2012). The
dopaminergic differentiation of SHED has been reported
previously, and the engraftment of dopaminergic neurons
obtained from SHED in the brain has been shown to
improve the production of dopamine and the abnormal
behaviors of Parkinsonian rats (Fujii et al., 2015; Wang et
al., 2010). Different protocols for the neuronal differentia-
tion of SHED have been established. Therefore, SHED are
promising stem cells for transplantation therapy, clinical
research, and drug discovery for neurological diseases. However, the mechanisms underlying the neuronal differ-
entiation of SHED remain unclear. Stem cells from human exfoliated deciduous teeth (SHED)
are isolated from the dental pulp tissue of primary teeth
(Miura et al., 2003). Exfoliated deciduous teeth are usually
discarded as medical waste; therefore, SHED can be easily
collected from exfoliated deciduous teeth with cooperation
and agreement from the patient. SHED express embryonic
stem cell markers Oct-4 and Nanog, the mesenchymal stem
cell marker stromal precursor antigen-1 (STRO-1), and the
neuronal stem cell marker Nestin and possess high prolifer- Mitochondria play a central role in ATP production, cel-
lular Ca2+ buffering, and apoptosis. Additionally, mitochon-
drial DNA (mtDNA) mutations induce aberrations in
oxidative phosphorylation (Keogh and Chinnery, 2015), 105 H. Kato et al. culture (Greiner Bio-one, Frickenhausen, Germany). After 24 h,
spheres were observed under an IX41 microscope (Olympus,
Tokyo, Japan) equipped with a digital camera NEX-5T (Sony,
Tokyo, Japan) with an NY-1S adaptor (MeCan Imaging, Saitama,
Japan). and embryonic stem cells (ESCs) harboring mtDNA muta-
tions have been reported to show reduced neuronal differ-
entiation (Kirby et al., 2009). Inhibition of mitochondrial
complex III, a complex required for oxidative phosphoryla-
tion, blocks the neuronal differentiation of ESCs (Pereira et
al., 2013). Furthermore, inhibition of mitochondrial com-
plex III results in high expression of Oct-4 during the neu-
ronal differentiation of ESCs (Pereira et al., 2013). These
results suggest that mitochondrial activity is involved in the
neuronal differentiation of stem cells. Measurement of mitochondrial membrane potential JC-1 is a cationic dye that accumulates in the mitochondria
depending on the MMP and forms aggregates. JC-1 shows a fluo-
rescence emission shift from green (monomer; 527 nm) to red
(aggregate; 590 nm) after forming aggregates (Reers et al., 1991). The MMP can therefore be measured by determining the ratio of
red to green fluorescent signals. SHED were incubated with 1 μM
JC-1 (Life Technologies) for 10 min and then washed three times
with PBS. The cells were then treated with Tryple Express (Life
Technologies) to detach them from the culture plates and collected
in tubes. Next, the cells were centrifuged at 800×g for 5 min, and
the collected cells were resuspended in PBS. JC-1 aggregates and
monomers were measured using a FACSCalibur (BD Bioscience,
CA, USA). Populations of 10,000 cells were analyzed for each
sample, and JC-1 signals were detected using the FL2 and FL1
channels for aggregates and monomers, respectively. Geometric
means of red and green fluorescence were measured using Cell
Quest software (BD Bioscience), and the ratio of red/green fluo-
rescence was calculated. Measurement of living cell numbers and the
percentages of living cells A total of 5×104 SHED were seeded into 6-well plates (Corning). After 24 h, SHED were treated with rotenone (Sigma-Aldrich; 1–
1,000 nM) and CCCP (Wako Pure Chemical Industries; 2.5–50
μM) for 2 days. The cells were then collected by trypsinization
and stained with 0.4% trypan blue solution (Wako Pure Chemical
Industries). The cells were counted with a hemocytometer, and
living and dead cells were distinguished by trypan blue staining. To calculate the percentage of living cells, the living cells were
divided by the total cell count (living cells and dead cells). To observe JC-1 aggregates and monomers using confocal
microscopy, SHED were cultured on glass-bottomed dishes (ibidi,
Munich, Germany). SHED were then incubated with 1 μM JC-1
(Life Technologies) for 10 min, and the culture medium was
changed. Fluorescent images of JC-1 aggregates and monomers
were acquired using a C2 confocal microscope (Nikon, Tokyo,
Japan). Western blotting The cells were washed three times with PBS and then lysed with
sample buffer (62.5 mM Tris-HCl buffer [pH 6.8] containing 2%
sodium dodecyl sulfate [SDS], 5% β-mercaptoethanol, and 10%
glycerol). The cell lysates were incubated for 95°C and analyzed
by SDS-polyacrylamide gel electrophoresis (PAGE). Subse-
quently, immunoblotting was performed using anti-β-tubulin III
(Sigma-Aldrich), anti-Nestin (Millipore), anti-Tom20 (Santa Cruz 106 Mitochondria Are Involved in SHED Differentiation genes were analyzed using the comparative the threshold cycle
(CT) method by normalizing to 18S rRNA. Biotechnology), and anti-heat shock protein 90 (HSP90; Santa
Cruz Biotechnology) antibodies. The immunoreactive bands were
detected using ECL Prime (GE Healthcare, Buckinghamshire,
UK) and analyzed using LAS-1000 pro (Fuji Film, Tokyo, Japan)
and Image Gauge software (Fuji Film). RNA extraction and quantitative real-time polymerase
chain reaction (PCR) A total of 5×105 SHED were seeded into 6-well plates (Corning). After 24 h, SHED were treated with DMSO, 100 nM rotenone
(Sigma-Aldrich), or 100 nM rotenone with 5 mM N-acetyl-L-
cysteine (NAC; Sigma-Aldrich) for 2 days. Then, the cells were
incubated with 5 μM MitoSOX Red (Life Technologies) for 10
min and washed three times with PBS. The cells were then treated
with Tryple Express (Life Technologies) to detach them from the
culture plates and collected in tubes. Next, the cells were centri-
fuged at 800×g for 5 min, and the collected cells were resuspen-
ded in PBS. The fluorescent signal of MitoSOX Red was
measured using a FACSCalibur (BD Bioscience). Total RNA was extracted from the cells using an RNAeasy Mini
Kit (Qiagen, Hilden, Germany). First-strand cDNA was synthe-
sized using a SuperScript III First Strand Synthesis System (Life
Technologies). The sequences of primer sets used in this study
were as follows: β-tubulin III, 5'-CTCAGGGGCCTTTGGA-
CATC-3' (β-tubulin III forward) and 5'-CAGGCAGTCGCAG-
TTTTCAC-3' (β-tubulin III reverse); postsynaptic density protein
95 (PSD95), 5'-TCGGTGACGACCCATCCAT-3' (PSD95 for-
ward) and 5'-GCACGTCCACTTCATTTACAAAC-3' (PSD95
reverse); gamma-aminobutyric acid (GABA) receptor α2, 5'-
GCCAATCAATCGGAAAGGAGAC-3' (GABA receptor α2
forward) and 5'-TTCCCATCCCAAGCCCATCCTC-3' (GABA
receptor α2 reverse); and 18S rRNA, 5'-CGGCTACCACATC-
CAAGGAA-3' (18S rRNA forward) and 5'-GCTGGAATTACC-
GCGGCT-3' (18S rRNA reverse). Real-time quantitative PCR
was performed using GoTaq qPCR Master Mix (Promega, WI,
USA) and analyzed with StepOnePlus Real-Time PCR Systems
(Life Technologies). The relative expression levels of the target Measurement of mtDNA amount by quantitative real-
time PCR mtDNA and nuclear DNA were extracted from the cells using a
QIAamp DNA mini kit (Qiagen). Mitochondrial tRNALEU(UUR) was
detected using the following primer set: 5'-CACCCAAGAA-
CAGGGTTTGT-3' (tRNA F3212) and 5'-TGGCCATGGGTA-
TGTTGTTA-3' (tRNA R3319) (Venegas and Halberg, 2012). Nuclear β-2-microglobulin (β2M) was detected using the primer
set 5'-TGCTGTCTCCATGTTTGATGTATCT-3' (β2M F594) and
5'-TCTCTGCTCCCCACCTCTAAGT-3' (β2M R679) (Venegas
and Halberg, 2012). Real-time PCR was performed as described
for mRNA measurement. Analysis of the relative amount of
mtDNA was performed as described previously (Venegas and
Halberg, 2012). Differentiation of SHED into neuronal cells Next, we examined the neuronal differentiation capacity of
the cultured SHED. SHED were cultured in neuronal differ-
entiation medium (Miura et al., 2003) for 10 days. Undif-
ferentiated and neuronally differentiated SHED were
stained with antibodies against the neuronal cell marker
proteins. SHED expressed β-tubulin III and MAP2 both
before (Fig. 2A, B) and after neuronal differentiation (Fig. 2C, D). After neuronal differentiation, elongation of the Statistical analysis Statistical analyses were performed with Wilcoxon tests using
JMP software (SAS Institute, NC, USA). Differences with P val-
ues of less than 0.05 were considered significant. 107 H. Kato et al. Results cells and neurite-like structures were observed (Fig. 2C, D;
indicated using white arrows). Moreover, the expression of
NMDAR1 was observed in neurite-like structures as puncta
after neuronal differentiation (Fig. 2E and F). Isolation and characterization of SHED Isolation and characterization of SHED To analyze the neuronal differentiation of SHED, SHED
were isolated from a primary tooth and were then cultured
until passage 5. The isolated SHED showed a fibroblast-
like morphology (Fig. 1A). To evaluate the stem cell prop-
erties of these SHED, we examined sphere formation
ability (Pastrana et al., 2011) and stem cell marker expres-
sion. SHED formed spheres when cultured under nonadher-
ent conditions (Fig. 1B). SHED expressed STRO-1 (a
mesenchymal stem cell marker, Fig. 1C). Moreover,
although there were differences in the expression levels
among the different cells (Fig. 1E; white arrow indicates
strong expression), all SHED expressed Nestin (a neuronal
stem cell marker, Fig. 1D). These results indicated that
SHED isolated in this study possessed stem cell character-
istics, as described previously (Miura et al., 2003). Next, the expression of the neuronal stem cell and neuro-
nal cell marker proteins β-tubulin III and Nestin were
examined using western blotting. After neuronal differen-
tiation, the expression levels of β-tubulin III and Nestin
were increased in SHED (Fig. 3A–C). In addition, real-time
PCR analysis showed that the expression levels of the neu-
ronal cell markers β-tubulin III (Fig. 3D), PSD95 (Fig. 3E),
and GABA receptor α2 (Fig. 3F) were increased after neu-
ronal differentiation of SHED. These increases in neuronal
cell markers indicated that SHED differentiated into neuro-
nal cells. In addition, Nestin is expressed during the early
stages of neuronal differentiation, e.g., neuronal stem cells,
progenitors, and immature neuronal cells (Von Bohlen Und
Halbach, 2007). Thus, the increase in Nestin expression
after neuronal differentiation suggested that neuronally dif-
ferentiated SHED in this study were in the early stage of
neuronal differentiation and were still progressing towards
neuronal linage differentiation. Increased mitochondrial activity during neuronal
differentiation of SHED To analyze mitochondrial activity during the neuronal dif-
ferentiation of SHED, MMP was measured by staining with
JC-1. Undifferentiated and neuronally differentiated SHED
were incubated with JC-1, and the JC-1 aggregates (red) Fig. 1. Isolation and characterization of SHED from human exfoliated deciduous teeth. (A) Isolated SHED were observed under a phase-contrast
microscope. Scale bar=50 μm. (B) SHED were cultured on 6-well plates for suspension cell culture for 24 h. Scale bar=50 μm. (C) SHED were stained
with DAPI and anti-STRO-1 antibodies. Scale bar=25 μm. (D) SHED were stained with DAPI and anti-Nestin antibodies. Scale bar=100 μm. (E) Details
of the boxed region in (D) are shown. Scale bar=25 μm. Fig. 1. Isolation and characterization of SHED from human exfoliated deciduous teeth. (A) Isolated SHED were observed under a phase-contrast
microscope. Scale bar=50 μm. (B) SHED were cultured on 6-well plates for suspension cell culture for 24 h. Scale bar=50 μm. (C) SHED were stained
with DAPI and anti-STRO-1 antibodies. Scale bar=25 μm. (D) SHED were stained with DAPI and anti-Nestin antibodies. Scale bar=100 μm. (E) Details
of the boxed region in (D) are shown. Scale bar=25 μm. 108 Mitochondria Are Involved in SHED Differentiation Fig. 2. Neuronal differentiation of SHED. (A, B) Undifferentiated SHED were stained with anti-β-tubulin III (A) and anti-MAP2 (B) antibodies and
counterstained with DAPI. Scale bar=50 μm. (C–F) SHED were differentiated into neuronal cells for 10 days and were then stained with anti-β-tubulin III
(C), anti-MAP2 (D), and anti-NMDAR1 (E) antibodies and counterstained with DAPI. Neurite-like structures are indicated by white arrows. Scale bar=50
μm. (F) Details of the boxed region in (E) are shown. Scale bar=10 μm. Fig. 2. Neuronal differentiation of SHED. (A, B) Undifferentiated SHED were stained with anti-β-tubulin III (A) and anti-MAP2 (B) antibodies and
counterstained with DAPI. Scale bar=50 μm. (C–F) SHED were differentiated into neuronal cells for 10 days and were then stained with anti-β-tubulin III
(C), anti-MAP2 (D), and anti-NMDAR1 (E) antibodies and counterstained with DAPI. Neurite-like structures are indicated by white arrows. Scale bar=50
μm. (F) Details of the boxed region in (E) are shown. Scale bar=10 μm. mitochondrial mass, we measured the amount of Tom20,
which is often used to measure mitochondrial mass as a
mitochondrial marker protein (Bueno et al., 2015; Burbulla
et al., 2014). There were no significant differences in the
amount of Tom20 before and after neuronal differentiation
(Fig. Increased mitochondrial activity during neuronal
differentiation of SHED (A) Total cell lysates were analyzed by SDS-PAGE with subsequent immunoblotting using
anti-β-tubulin III (upper panel), anti-Nestin (middle panel) and anti-HSP90 (lower panel; as an internal control) antibodies. (B, C) The band intensities of
(A) were quantified. The relative expression of β-tubulin III (B) and Nestin (C) protein was normalized to HSP90 protein levels. The means±SEMs from
four experiments are shown in the graph. *P<0.05. (D–F) The mRNA expression levels of the neuronal cell markers β-tubulin III (D), PSD95 (E), and
GABA receptor α2 (F) were measured using real-time PCR. The relative expression of neuronal cell markers was normalized to the expression of 18S
rRNA. The means±SEMs from four experiments are shown in the graph. *P<0.05. Fig. 3. Expression of neuronal markers during neuronal differentiation of SHED. Total cell lysates and RNA were prepared from undifferentiated SHED
and SHED cultured for neuronal differentiation for 10 days. (A) Total cell lysates were analyzed by SDS-PAGE with subsequent immunoblotting using
anti-β-tubulin III (upper panel), anti-Nestin (middle panel) and anti-HSP90 (lower panel; as an internal control) antibodies. (B, C) The band intensities of
(A) were quantified. The relative expression of β-tubulin III (B) and Nestin (C) protein was normalized to HSP90 protein levels. The means±SEMs from
four experiments are shown in the graph. *P<0.05. (D–F) The mRNA expression levels of the neuronal cell markers β-tubulin III (D), PSD95 (E), and
GABA receptor α2 (F) were measured using real-time PCR. The relative expression of neuronal cell markers was normalized to the expression of 18S
rRNA. The means±SEMs from four experiments are shown in the graph. *P<0.05. Increased mitochondrial activity during neuronal
differentiation of SHED 5C), suggesting that mitochondrial mass was not
changed during the neuronal differentiation of SHED. Moreover, we examined morphology of mitochondria dur-
ing neuronal differentiation of SHED (Fig. 5D–F). After
neuronal differentiation, elongated mitochondria were
observed in neurite-like structures (Fig. 5E, F; indicated
using white arrows). These results indicated that increased
MMP and mtDNA and elongated mitochondria were
involved in the neuronal differentiation of SHED. and monomers (green) were detected by confocal micro-
scopy. After neuronal differentiation for 2 days, the JC-1
aggregates (red) increased when compare with that before
neuronal differentiation (Fig. 4A and B, left panel). Fur-
thermore, we analyzed the ratio of JC-1 aggregates/mono-
mers using flow cytometry. The ratio of JC-1 aggregates/
monomers was increased by 1.87-fold after neuronal differ-
entiation for 2 days (Fig. 4C–E). Moreover, after neuronal
differentiation for 10 days, the ratio of JC-1 aggregates/
monomers was increased by 2.88-fold compared with that
before neuronal differentiation (Fig. 4F). Next, we examined the factors involved in mitochondrial
activity, mtDNA contents, and mitochondrial mass and
morphology during neuronal differentiation of SHED. After neuronal differentiation for 2 or 10 days, the amount
of mtDNA was increased (Fig. 5A, B). Next, to evaluate 109 H. Kato et al. Fig. 3. Expression of neuronal markers during neuronal differentiation of SHED. Total cell lysates and RNA were prepared from undifferentiated SHED
and SHED cultured for neuronal differentiation for 10 days. (A) Total cell lysates were analyzed by SDS-PAGE with subsequent immunoblotting using
anti-β-tubulin III (upper panel), anti-Nestin (middle panel) and anti-HSP90 (lower panel; as an internal control) antibodies. (B, C) The band intensities of
(A) were quantified. The relative expression of β-tubulin III (B) and Nestin (C) protein was normalized to HSP90 protein levels. The means±SEMs from
four experiments are shown in the graph. *P<0.05. (D–F) The mRNA expression levels of the neuronal cell markers β-tubulin III (D), PSD95 (E), and
GABA receptor α2 (F) were measured using real-time PCR. The relative expression of neuronal cell markers was normalized to the expression of 18S
rRNA. The means±SEMs from four experiments are shown in the graph. *P<0.05. Fig. 3. Expression of neuronal markers during neuronal differentiation of SHED. Total cell lysates and RNA were prepared from undifferentiated SHED
and SHED cultured for neuronal differentiation for 10 days. Inhibition of neuronal differentiation by inhibition of
mitochondrial activity was then measured using JC-1. Notably, 100 nM rotenone
and 7.5 μM CCCP markedly reduced MMP during the neu-
ronal differentiation of SHED (Fig. 6E, F). In contrast, 5
μM CCCP did not reduced the MMP (Fig. 6F). Thus, these
data showed that 100 nM rotenone and 7.5 μM CCCP
could be used without substantially reducing cell growth
and viability; accordingly, we used 100 nM rotenone and
7.5 μM CCCP for further experiments. To evaluate the involvement of mitochondrial activity in
neuronal differentiation, SHED were treated with rotenone,
a mitochondrial complex I inhibitor, and CCCP, a mito-
chondrial uncoupler, during their neuronal induction. To
examine the effects of rotenone and CCCP without reduc-
ing cell growth and viability, we examined the
concentration-dependent toxicities of rotenone and CCCP
in SHED. SHED were cultured in the presence or absence
(DMSO) of rotenone or CCCP for 2 days, and the numbers
and percentages of living cells were then measured. In the
presence of 1,000 nM rotenone and more than 7.5 μM
CCCP, the number of living cells was significantly reduced
(Fig. 6A, C). In addition, 1,000 nM rotenone and 50 μM
CCCP significantly reduced the percentages of living cells
(Fig. 6B, D). Next, we evaluated the inhibitory effects of
rotenone and CCCP on the MMP under neuronal differen-
tiation conditions. SHED were cultured in neuronal differ-
entiation medium in the presence or absence of 100 nM
rotenone and 5 and 7.5 μM CCCP for 2 days, and the MMP SHED were differentiated into neuronal cells in the pres-
ence or absence (DMSO) of 100 nM rotenone and 7.5 μM
CCCP for 10 days. The generation of neurite-like structures
was reduced in the presence of rotenone and CCCP com-
pared with that observed in the absence of these chemicals
(Fig. 7A–C; white arrows). Furthermore, measurement of
cell length revealed that the elongation of cells was inhibi-
ted by rotenone and CCCP (Fig. 7E), and the expression of
β-tubulin III and Nestin was reduced in the presence of
rotenone and CCCP (Fig. 7F–H). Because treatment with
7.5 μM CCCP resulted in reduction of number of cells after
2 days of culture (Fig. 6C), we measured the cell numbers
after neuronal differentiation for 10 days in the presence or
absence (DMSO) of 7.5 μM CCCP. Although the cell num- 110 Mitochondria Are Involved in SHED Differentiation Fig. 4. Inhibition of neuronal differentiation by inhibition of
mitochondrial activity Increased mitochondrial membrane potential during the neuronal differentiation of SHED. (A, B) Undifferentiated SHED (A) and SHED cultured
for neuronal differentiation for 2 days (B) were incubated with JC-1. JC-1 red (aggregates; left panel) and green (monomers; middle panel) were observed
with confocal microscopy. Merged images are shown in the right panel. Scale bar=20 μm. (C–E) Undifferentiated SHED (C) and SHED cultured for
neuronal differentiation for 2 days (D) were used for measurement of MMP by JC-1 staining. JC-1 red (left panel) and green (right panel) were analyzed
by flow cytometry. The relative differences in the ratio of red/green fluorescence were calculated, and the ratio of red/green fluorescence in the
undifferentiated SHED was set as 1. The means±SEMs from four experiments are shown in the graph (E). *P<0.05. (F) SHED were differentiated into
neuronal cells for 10 days, and the MMP was measured as described above. The means±SEMs from four experiments are shown in the graph. *P<0.05. Fig. 4. Increased mitochondrial membrane potential during the neuronal differentiation of SHED. (A, B) Undifferentiated SHED (A) and SHED cultured
for neuronal differentiation for 2 days (B) were incubated with JC-1. JC-1 red (aggregates; left panel) and green (monomers; middle panel) were observed
with confocal microscopy. Merged images are shown in the right panel. Scale bar=20 μm. (C–E) Undifferentiated SHED (C) and SHED cultured for
neuronal differentiation for 2 days (D) were used for measurement of MMP by JC-1 staining. JC-1 red (left panel) and green (right panel) were analyzed
by flow cytometry. The relative differences in the ratio of red/green fluorescence were calculated, and the ratio of red/green fluorescence in the
undifferentiated SHED was set as 1. The means±SEMs from four experiments are shown in the graph (E). *P<0.05. (F) SHED were differentiated into
neuronal cells for 10 days, and the MMP was measured as described above. The means±SEMs from four experiments are shown in the graph. *P<0.05. Rotenone increases the production of mtROS (Cadenas
et al., 1977), and mtROS production reduces the neuronal
differentiation of ESCs (Pereira et al., 2013). mtROS pro-
duction increased when SHED were cultured with rotenone
(Fig. 7I). To prevent the effects of mtROS on neuronal dif-
ferentiation, we examined the role of the antioxidant NAC. NAC significantly reduced mtROS production increased by
rotenone (Fig. 7I). Inhibition of neuronal differentiation by inhibition of
mitochondrial activity However, when SHED were differenti-
ated into neuronal cells with rotenone and NAC, the reduc- bers showed reduction after treatment with 7.5 μM CCCP
(Fig. 6G; 73.0%; compared with DMSO control), similar to
our observation after 2 days of culture (71.7%; compared
with DMSO and 7.5 μM CCCP treatment in Fig. 6C), no
significant difference was observed in the cell numbers
between the DMSO and 7.5 μM CCCP treated cells
(P=0.12). Accordingly, these results showed that inhibition
of mitochondrial activity by rotenone and CCCP reduced
the neuronal differentiation of SHED. 111 H. Kato et al. Fig. 5. Increased mtDNA and elongation of mitochondria during the neuronal differentiation of SHED. (A, B) mtDNA and nuclear DNA were isolated
from SHED differentiated for 2 (A) or 10 days (B). The relative amount of mtDNA was analyzed by real-time PCR. The means±SEMs from four
experiments are shown in the graph. *P<0.05. (C) Total cell lysates were prepared from undifferentiated SHED and SHED cultured for neuronal
differentiation for 10 days. Then, the lysates were analyzed by western blotting using anti-Tom20 antibodies. The relative expression of Tom20 was
normalized to HSP90 protein levels. The means±SEMs from four experiments are shown in the graph. NS, not significant. (D, E) Undifferentiated SHED
(D) and SHED cultured for neuronal differentiation for 10 days (E) were stained with anti-Tom20 (left panel) and anti-β-tubulin III (middle panel)
antibodies and counterstained with DAPI. Merged images are shown in the right panel. Scale bar=20 μm. (F) Detailed mitochondrial morphology of the
boxed region in (E) is shown. Scale bar=20 μm. Fig. 5. Increased mtDNA and elongation of mitochondria during the neuronal differentiation of SHED. (A, B) mtDNA and nuclear DNA were isolated
from SHED differentiated for 2 (A) or 10 days (B). The relative amount of mtDNA was analyzed by real-time PCR. The means±SEMs from four
experiments are shown in the graph. *P<0.05. (C) Total cell lysates were prepared from undifferentiated SHED and SHED cultured for neuronal
differentiation for 10 days. Then, the lysates were analyzed by western blotting using anti-Tom20 antibodies. The relative expression of Tom20 was
normalized to HSP90 protein levels. The means±SEMs from four experiments are shown in the graph. NS, not significant. (D, E) Undifferentiated SHED
(D) and SHED cultured for neuronal differentiation for 10 days (E) were stained with anti-Tom20 (left panel) and anti-β-tubulin III (middle panel)
antibodies and counterstained with DAPI. Inhibition of neuronal differentiation by inhibition of
mitochondrial activity Merged images are shown in the right panel. Scale bar=20 μm. (F) Detailed mitochondrial morphology of the
boxed region in (E) is shown. Scale bar=20 μm. chondria in the neuronal differentiation of SHED. The
amount of mtDNA was increased during neuronal differen-
tiation. mtDNA contains 37 genes, 13 of which are
involved in mitochondrial oxidative phosphorylation
(Anderson et al., 1981). The amount of mtDNA is correla-
ted with mitochondrial respiratory activity and changes
depending on energy requirements (D’Erchia et al., 2015). Our results showed that the mass of the mitochondria did
not change during neuronal differentiation, suggesting that
mtDNA content in each mitochondrion was increased and
that mitochondrial function was increased during neuronal tion of neurite-like structures and elongation of the cells
were observed, similar to rotenone treatment alone (Fig. 7D, E). This result indicated that the inhibitory effects of
rotenone on neuronal induction of SHED were not caused
by mtROS, but were instead caused by reduction of mito-
chondrial activity. Discussion In this study, we demonstrated the involvement of mito- 112 Mitochondria Are Involved in SHED Differentiation Fig. 6. Concentration-dependent toxicities of rotenone and CCCP on SHED. (A–D) SHED were treated with rotenone (1–1,000 nM) and CCCP (2.5–50
μM) for 2 days. The cells were then stained with trypan blue solution to distinguish live and dead cells. The number of living cells is shown in (A, C), and
the percentage of living cells is shown in (B, D). The means±SEMs from four experiments are shown in the graph. *P<0.05. (E, F) SHED were
differentiated into neuronal cells for 2 days in the presence or absence of 100 nM rotenone (E) and 5 or 7.5 μM CCCP (F), and the MMP was then
measured as described in Fig. 4. The means±SEM from four experiments are shown in the graph. *P<0.05. NS, not significant. (G) The cell numbers were
measured after neuronal differentiation for 10 days in the presence or absence of 7.5 μM CCCP. The relative differences in the cell numbers were
calculated, and the DMSO control was set as 100%. Results are shown as the mean±SEM of five experiments. NS, not significant. Fig. 6. Concentration-dependent toxicities of rotenone and CCCP on SHED. (A–D) SHED were treated with rotenone (1–1,000 nM) and CCCP (2.5–50
μM) for 2 days. The cells were then stained with trypan blue solution to distinguish live and dead cells. The number of living cells is shown in (A, C), and
the percentage of living cells is shown in (B, D). The means±SEMs from four experiments are shown in the graph. *P<0.05. (E, F) SHED were
differentiated into neuronal cells for 2 days in the presence or absence of 100 nM rotenone (E) and 5 or 7.5 μM CCCP (F), and the MMP was then
measured as described in Fig. 4. The means±SEM from four experiments are shown in the graph. *P<0.05. NS, not significant. (G) The cell numbers were
measured after neuronal differentiation for 10 days in the presence or absence of 7.5 μM CCCP. The relative differences in the cell numbers were
calculated, and the DMSO control was set as 100%. Results are shown as the mean±SEM of five experiments. NS, not significant. differentiation of SHED. neurites are important for neuronal development of SHED. p
p
We showed that mitochondrial activity was increased
during the neuronal differentiation of SHED and that inhib-
ition of mitochondrial activity delayed neuronal differentia-
tion. Our results were similar to the results of previous
studies of the neuronal differentiation of ESCs. Similar to
SHED, ESCs also exhibit increased mitochondrial activity
during differentiation (Pereira et al., 2013). Furthermore,
the inhibition of mitochondrial activity also reduces the
neuronal differentiation of ESCs (Pereira et al., 2013). Interestingly, to maintain their stemness, ESCs harbor a
limited number of mitochondria and reduce their mitochon-
drial activity before differentiation (Armstrong et al., 2010;
Chen et al., 2012). The mitochondrion is a major site of
ROS generation due to oxidative phosphorylation, and high
levels of ROS damage both mitochondrial and genomic Mitochondrial morphology is highly diversified and is
correlated with mitochondrial activity. Elongated mitochon-
dria possess high mitochondrial respiratory capacity, and
the opposite is also true (Picard et al., 2013). In the present
study, elongated mitochondria were observed in neurite-
like structures after neuronal differentiation of SHED. Mitochondrial localization in axons and local ATP produc-
tion were required for the events of neuronal outgrowth,
including growth corn motility and cytoskeletal assembly
(Bernstein and Bamburg, 2003; Morris and Hollenbeck,
1993; Ruthel and Hollenbeck, 2003; Vaarmann et al.,
2016). In addition, elongated mitochondria have been
observed in the neurites of cortical neurons during neuronal
differentiation (Cho et al., 2014). Thus, our results sugges-
ted that localization and elongation of mitochondrial in 113 H. Kato et al. ory effect of rotenone and CCCP on neuronal differentiation of SHED. (A–D) SHED were differentiated into neuron
absence (DMSO; A) of 100 nM rotenone (B), 7.5 μM CCCP (C), and 100 nM rotenone with 5 mM NAC (D). The c
anti-β-tubulin III antibodies. Neurite-like structures are indicated with white arrows. Scale bar=100 μm. (E) The cell l
dot plots show the lengths of 250 cells from five experiments, and the black bars show the means. *P<0.01. NS, not
e prepared and analyzed by SDS-PAGE with subsequent immunoblotting using anti-β-tubulin III (upper panel), anti-N
(lower panel) antibodies. (G, H) Band intensities of (F) were quantified. The relative expression levels of β-tubulin I
rmalized to HSP90 protein levels, and the means±SEMs from four experiments are shown in the graph. *P<0.05. differentiation of SHED. Accordingly, we speculate that SHED may also
reduce their mitochondrial activity before differentiation to
avoid DNA damage from high levels of ROS in order to
maintaining their stemness. Armstrong, L., Tilgner, K., Saretzki, G., Atkinson, S.P., Stojkovic, M.,
Moreno, R., Przyborski, S., and Lako, M. 2010. Human induced pluri-
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2013). In contrast, lactate levels are increased during this
period (Prigione et al., 2010). In addition, inhibition of
mitochondrial activity by rotenone enhances the reprogram-
ming of somatic cells into iPSCs (Son et al., 2013), and the
inhibition of mitochondrial activity by antimycine A
increases Oct-4 expression (Pereira et al., 2013). Taken
together, these data indicate that low levels of mitochon-
drial activity tend to maintain stem cells in the undifferenti-
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heteroplasmic mutation 3380G>A in ND1 of mtDNA. Neuromuscul. Disord., 18: 553–556. Acknowledgments. We thank Drs. Y. Ninomiya, H. Nakanishi, T. Yamaza, and K. Sanematsu and all members of the Pediatric & Special
Needs Dentistry at Kyushu University Hospital for the valuable
suggestions, technical support, and materials. differentiation of SHED. We appreciate the technical
assistance provided by the Research Support Center, Research Center for
Human Disease Modeling, Kyushu University Graduate School of
Medical Sciences. This work was supported by JSPS KAKENHI (grant
numbers 25670877 and 16K15839). The authors deny any conflicts of
interest related to this study. Jarmalavičiūtė,
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R393-406. Yamaza, T., Alatas, F.S., Yuniartha, R., Yamaza, H., Fujiyoshi, J.K.,
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hepatogenic capacity and therapeutic potential of stem cells from
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8618–8624. 116
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Notes on the Birds of the Cariboo District, British Columbia
|
Allan Brooks
|
English
|
Spoken
| 3,251
| 8,296
|
ee
Brooks,
Birds
of
the
Cariboo
District,
B. ee
Brooks,
Birds
of
the
Cariboo
District,
B. 277 Plate X. DEsIRING to study the fauna of the northern interior of British
Columbia,
I
spent
fifteen
months,
from
June,
1g00,
to
October,
1go1, in the Cariboo district, a large portion of the time being
devoted
entirely
to
collecting. The
first
eleven
months
were
spent in the heavily timbered country in the northern portion of
the
district,
Quesnelle
Mouth,
Willow
River,
and
the
mountains
southeast
of
Barkerville. -From
May,
rgor,
till
the
following
October I made my headquarters at the 158-Mile House, in the
southwestern corner of the district, just north of the 52d parallel. Excursions were made from this point to the Chilcotin plateau,
Lac la Hache and Horsefly River. The country around Quesnelle, on both banks of the Fraser,
and north to Fort George, is entirely covered with forest, mostly
coniferous, — spruce, balsam, Murray pine and Douglas fir, with
a good deal of birch and poplar. The altitude of the Fraser at
Quesnelle is 1600 feet. The mountains in this region are mostly low, level plateaus, but
towards Barkerville they merge into the Cariboo Range, rugged and
snow-capped, with timber line at about 5500 feet. Towards the southern portion of this range the climate becomes
more humid and the valleys, such as the upper Horsefly, possess a
forest growth very similar to that of the coast region, — hemlock,
cedar, Douglas fir, yew, etc., with a heavy growth of underbrush,
red dogwood, devil’s club, etc. The
southwestern
corner
of
the
Cariboo
district,
like
the
Lilloet district to the southwest, is diversified with a good deal of
open
and
partially
timbered
country;
the
158-Mile
House
is
situated
on
a
plateau
of
about
3000
feet
altitude
(Carpenter’s
Mountain). Here
there
is
a
good
deal
of
natural
prairie,
with
numerous lakes and ponds, and scattered groves of timber and
brush, the fauna and flora having many of the characteristics of
the plains to the east of the Rockies. 278 rat ks,
Birds
of
the
Cariboo
District,
B. The whole district has a very cold winter climate and a moder-
ately warm summer. Mosquitoes and blackflies swarm, and _ bird-
nesting in the swamps and woods is generally anything but a
pleasure. 1. Colymbus
holbellii. Horpa:ii’s
GREBE. 1. Colymbus
holbellii. Horpa:ii’s
GREBE. 2. Colymbus auritus. HoRNED GREBE. 2. Colymbus auritus. HoRNED GREBE. Both these grebes were abundant, breeding on nearly every pond and
lake. Plate X. ‘The larger species wages incessant war upon the smaller one, the
larger birds diving and coming up beneath the smaller ones time and
again to the terror of the poor little fellows, who often desert their nests
in consequence. q
3. Podilymbus podiceps. _PIED-BILLED GREBE.— Rare. 3. Podilymbus podiceps. _PIED BILLED GREBE. Rare. I kept a good lookout for the Western Grebe (<&chmophorus occi-
dentalis), but never saw one, not even during migrations. Their line of
migration is probably straight eastward from southern British Columbia,
where they are common. 4. Gavia
imber. Loon.—
Abundant;
the
only
species
of
loon
observed. 5. Larus philadelphia. BONAPARTE’s GuLL.— The only gull observed
during the breeding season. Breeds in the netghborhood of Quesnelle
Lake. 6. Merganser
americanus. AMERICAN
MERGANSER.—
Breeding
on the streams and the larger lakes but absent from the smaller lakes
that are devoid of fish. 7. Lophodytes cucullatus. HooprEp MERGANSER.— Scarce. 7. Lophodytes cucullatus. HooprEp MERGANSER.— Scarce. b
h 8. Anas boschas. MALLARD. 8. Anas boschas. MALLARD. 8. Anas boschas. MALLARD. g. Mareca americana. BALDPATE. g. Mareca americana. BALDPATE. 10. Nettion carolinensis. GREEN-WINGED TEAL. These three species are all abundant breeders. ‘ ese three species are all abundant breeders. ‘tr. Querquedula discors. BLUE-WwINGED TEAL. 12. Spatula clypeata. SHOVELLER. 12. Spatula clypeata. SHOVELLER. 13. Dafila acuta. PINTAIL. These three ducks are rather scarce breeders in the neighborhood of
158-Mile
House. = These three ducks are rather scarce breeders in the neighborhood of
158-Mile
House. = These three ducks are rather scarce breeders in the neighborhood of
158-Mile
House. = I did not observe the Gadwall, the Cinnamon Teal, nor the Redhead,
which are probably not found north of Lac la Hache. 14. Aythya
vallisneria. CANVAs-BACK.—Common
breeder. The
nests are bulky platforms of reeds, similar to a Coot’s, found generally
on small swampy ponds, away from the larger lakes, where the males
associate in flocks. Eggs were taken from 21st of May to 6th June. 14. Aythya
vallisneria. CANVAs-BACK.—Common
breeder. The
nests are bulky platforms of reeds, similar to a Coot’s, found generally
on small swampy ponds, away from the larger lakes, where the males
associate in flocks. Eggs were taken from 21st of May to 6th June. 14. Aythya
vallisneria. CANVAs-BACK.—Common
breeder. The
nests are bulky platforms of reeds, similar to a Coot’s, found generally
on small swampy ponds, away from the larger lakes, where the males
associate in flocks. 1.
Charitonetta
albeola.
2.
Aythya
v
3.
Erismatura
jam Plate X. Eggs were taken from 21st of May to 6th June. 1s. Aythya
marila. Scaup
Ducx.—Observed
only
during
its
migrations. 1s. Aythya
marila. Scaup
Ducx.—Observed
only
during
its
migrations. g
16. Aythya affinis. Lress—ER Scaup Duck.— Abundant, breeding
much later than the Canvas-back or Ring-necked. The nests were g
16. Aythya affinis. Lress—ER Scaup Duck.— Abundant, breeding
much later than the Canvas-back or Ring-necked. The nests were Wiis
ING,
WO,
OR Pica OX. Pica OX. Allan
Wrocks
-
a5)
YOUNG DUCKS. 1. Charitonetta
albeola. 4. Glaucionetta
islandica. 2. Aythya
vallisneria. Aythya
collaris. un
3. Erismatura
jamaicensis. 6. Aythya
affinis. Allan
Wrocks
-
5) Allan
Wrocks
- 4. Glaucionetta
islandica. hya
collaris. un
Aythya
affinis. oks,
Birds
of
the
Cartboo
District,
B. 4 279 usually in coarse grass, with a waterway, generally a muskrat’s runway,
connecting with the nearest open water. Clutches varied from seven to
eleven eggs each. First eggs taken on 21st June. . 17. Aythya
collaris. RING-NECKED
Duck.—A_
rather
scarce
breeder. I was able to take only one set of eggs, evidently a second
laying, as there was no down. This was on the 27th June. The nest
was in a tussock of grass, in eight inches of water; it was composed of
coarse green grass and arched over with the drooping blades of the
tussock. The nine eggs contained small embryos. 17. Aythya
collaris. RING-NECKED
Duck.—A_
rather
scarce
breeder. I was able to take only one set of eggs, evidently a second
laying, as there was no down. This was on the 27th June. The nest
was in a tussock of grass, in eight inches of water; it was composed of
coarse green grass and arched over with the drooping blades of the
tussock. The nine eggs contained small embryos. Young broods of this species were observed before the Lesser Scaups
(A. affinis) had started to lay. The young in down are very light colored,
resembling the young of the Canvasback and Redhead, and quite dif-
ferent from the dusky, unspotted young of the Lesser Scaup. (See Pl. X.) 18. Clangula clangula americana. AMERICAN GOLDEN-EYE.— Com-
mon during migrations, but not observed during the breeding season. 19. Clangula islandica. BARROW’s GoLDEN-EYE.—A_ rather scarce
breeder in the neighborhood of 158-Mile House, but common in
La Hfiche Valley. Plate X. One set of eggs was taken from a hole in a dead
Douglas fir, fifty feet from the ground, probably the deserted nest of
a flying squirrel. The tree stood about four hundred yards from the
nearest water. The eggs (seven) at this date (17th June) contained large
embryos. I saw another nesting hole but was unable to reach it. The
female brought fourteen young ones out from this. 20. Charitonetta albeola. Burrire-1ieEAp.— Almost every ‘lake has
one or more pairs of these charming little ducks. Unlike Barrow’s
Goldeneye, the nests were always in trees close to, or but a short distance
away from water. These nests were invariably the deserted nesting holes
of flickers, and in most cases had been used several years in succession by
the ducks. The holes were in aspen trees, from five to twenty feet from
the ground, and the entrance was not more than three and a quarter
inches in diameter. The number of eggs ranged from two to nine, eight
being the average; in color they resemble old ivory, without any tinge
of green. I have several times seen the eggs of this duck described as
“dusky green,” but these have evidently been the eggs of some species
of Teal. The female Bufflehead is a very close sitter, never leaving the
nest until the hole was sawed out, and in most cases I had to lift the bird
and throw her up in the air, when she would make a bee-line for the
nearest lake, where her mate would be slowly swimming up and down
unconscious of the violation of his home. In many cases the eggs had
fine cracks, evidently made by the compression of the bird’s body when
entering the small aperture. 21. Hareldahyemalis. OLp-squAWw.— Common on the larger lakes, but
by June they had all gone North, with the exception of a single female
which remained on a small lake near the 158-Mile House throughout the
summer. 21. Hareldahyemalis. OLp-squAWw.— Common on the larger lakes, but
by June they had all gone North, with the exception of a single female
which remained on a small lake near the 158-Mile House throughout the
summer. 22. Histrionicus histrionicus. HARLEQUIN Duck. — A scarce summer
resident on some of the mountain streams. 22. Histrionicus histrionicus. HARLEQUIN Duck. — A scarce summer
resident on some of the mountain streams. All were taken at Quesnelle during migrations. 39. Calidris arenaria. SANDERLING. Plate X. [3a% 280 ks,
Birds
of
the
Cariboo
District,
B. 23. Oidemia deglandi. WHITE-wINGEpD ScoTER. — Numbers of these
scoters remained on the larger lakes near the 158-Mile House throughout
the summer, and to all appearances they were paired and breeding, yet I
never found a nest, nor saw any broods of young. 24. Oidemia
perspicillata. Surr
Scorer.—
Seen
throughout
the
summer but does not breed. 25. Erismatura
jamaicensis. Ruppy
Duck.—
A
common
breeder. While watching the curious antics of the males, through a binocular at
very close range, I was struck with the peculiar formation of the head,
there being distinct elevations over each eye resembling those of a frog. These were evidently caused by inflation from the inside of the skin. Young when first hatched are, as might be expected, very large, and dive
for their food, unlike all other young ducks, which take their food from
the surface for several weeks. 26. Branta
canadensis. CANADA
Goosr.—Common. This
is
the
only goose that breeds in Cariboo, Chilcotin, etc. I failed to find any
evidence of the breeding of Hutchins’s Goose, and all residents whom I
questioned asserted positively that they had never seen any of the smaller
geese breeding, though a few may remain through the summer, as they
do in southern British Columbia, evidently non-breeding birds. Many sets of the eggs of Canada Geese are taken and set under hens ;
these often produce undersized birds, which has led to the statement, so
often made, of the breeding of Hutchins’s Goose in British Columbia. Many sets of the eggs of Canada Geese are taken and set under hens ;
these often produce undersized birds, which has led to the statement, so
often made, of the breeding of Hutchins’s Goose in British Columbia. 27. Olor
buccinator. TRUMPETER
SWAN.—Swans
of
this
species
breed in northern Chilcotin. 27. Olor
buccinator. TRUMPETER
SWAN.—Swans
of
this
species
breed in northern Chilcotin. 28. Botaurus
lentiginosus. AMERICAN
BITTERN. southern portion of the district. Breeds in the 28. Botaurus
lentiginosus. AMERICAN
BITTERN. southern portion of the district. Breeds in the p
29. Grus mexicana. SANDHILL CRANE. — Breeds in suitable localities. The smaller species (G. canadensis) passes through on migrations only. 29. Grus mexicana. SANDHILL CRANE. — Breeds in suitable localities. The smaller species (G. canadensis) passes through on migrations only. o. Rallus virginianus. VIRGINIA RAIL. o. Rallus virginianus. VIRGINIA RAIL. 1. Porzana carolina. SORA. 36. Tringa minutilla. LEAST SANDPIPER.
a) 35. Tringa maculata. PECTORAL SANDPIPER. Plate X. OO
GW 2. Fulica americana. AMERICAN Coot. OO
GW 2. Fulica americana. AMERICAN Coot. OO
GW 2. Fulica americana. AMERICAN Coot. OO
GW All three are common and breed. 33. Phalaropus lobatus. NORTHERN PHALAROPE. — This phalarope
may breed in northern Chilcotin, though I could find no evidence of its
doing so near the 158 Mile House. g
34. Gallinago delicata. WuLson’s SNipE.— Common summer resi-
dent. 34. Gallinago delicata. WuLson’s SNipE.— Common summer resi-
dent. 35. Tringa maculata. PECTORAL SANDPIPER. 35. Tringa maculata. PECTORAL SANDPIPER. 36. Tringa minutilla. LEAST SANDPIPER. a) 36. Tringa minutilla. LEAST SANDPIPER. a) )
These two sandpipers were frequently seen throughout the summe),
but were evidently non-breeding birds. 37. Tringabairdii. BAIRD’s
SANDPIPER. 38. Ereunetes pusillus. SEMIPALMATED SANDPIPER. 38. Ereunetes pusillus. SEMIPALMATED SANDPIPER. 39. Calidris arenaria. SANDERLING. 39. Calidris arenaria. SANDERLING. All were taken at Quesnelle during migrations. i
ei 281 oks,
Birds
of
the
Cariboo
District,
B. 40. Totanus melanoleucus. GREATER YELLOW-LEGS.— Breeding in
many localities but all ettorts to find the nest were unsuccessful, owing to
the extreme watchfulness of the male bird, which kept constant watch
from the extreme summit of some tall spruce. Young were first observed
on 15th June. 40. Totanus melanoleucus. GREATER YELLOW-LEGS.— Breeding in
many localities but all ettorts to find the nest were unsuccessful, owing to
the extreme watchfulness of the male bird, which kept constant watch
from the extreme summit of some tall spruce. Young were first observed
on 15th June. 41. Helodromas solitarius cinnamomeus. WESTERN SOLITARY SAND-
PIPER.— I think the Solitary Sandpiper breeds in the district, as I took
young with the down still adhering to their plumage. y
g
g
p
g
42. Bartramia longicauda. BARTRAMIAN SANDPIPER. — Frequently
seen on both spring and autumn migrations. Mr. Sidney Williams took
one specimen at Quesnelle and I shot another at the 158-Mile House. p
43. Actitis macularia. SpoTreED SANDPIPER.— Common _ breeder. 44
Numenius
longirostris
LONG-BILLED
CuURLEW —
Breeding
in p
43. Actitis macularia. SpoTreED SANDPIPER.— Common _ breeder. 43. Actitis macularia. SpoTreED SANDPIPER.— Common _ breeder. 44. Numenius longirostris. LONG-BILLED CuURLEW.— Breeding in
the La Hache valley, but not observed in the Cariboo district proper. 44. Numenius longirostris. LONG-BILLED CuURLEW.— Breeding in
the La Hache valley, but not observed in the Cariboo district proper. 45. Squatarola squatarola. BLACK-BELLIED PLOVER. 46. Charadrius dominicus. AMERICAN GOLDEN PLOVER.— Seen only
during fall migrations. g
g
47. Plate X. &gialitis vocifera. KiILLDEER.— Common, breeds. 47. &gialitis vocifera. KiILLDEER.— Common, breeds. 48. Dendragapus
obscurus
richardsonii. RICHARDSON’S
GROUSE. —Common
in
partially
wooded
country
in
La
Hache
valley,
Chilcotin, and at Soda Creek, and again on the summits of the mountains
of the Cariboo Range, but not in the intervening heavily wooded country. All those secured showed faint traces of a terminal tail bar. 49. Camnachites franklinii. FRANKLIN’s GRousr.— Abundant in all
suitable localities. “Towards the northern portion of the district many
show an approach to typical caxadensts. pp
yp
50. Bonasa umbellus togata. CANADIAN RUFFED GROUSE. g
51. Bonasa umbellus umbelloides. GrRAy RuUFFED GROUSE. y
Most of the Ruffed Grouse of the district are intermediate between
these two races, but ultra-typical examples of each were taken. 52. Lagopus
leucurus. WHITE-TAILED
PTARMIGAN.—The
only
Ptarmigan observed. I could find no record of ZL. rufestris, although
it occurs further south. 53. Pedizcetes
phasianellus
columbianus. CoLUMBIAN
SHARP-
TAILED
GRousE.—
Abundant
at
158-Mile
House;
scarce
at
Ques-
nelle. Those taken at the latter locality show a close approach to
typical phasianellus. yp
p
54. Zenaidura macroura. MourninG Dove.— Scarce; in the south-
ern portion of the district only. 55. Nyctala
tengmalmi
=
richardsoni. RICHARDSON’S
OwL.—
Quesnelle. Q
56. Glaucidium
gnoma
californicum. CALIFORNIA
PyGmMy
OwL. — Taken as far north as Willow River. Q
56. Glaucidium
gnoma
californicum. CALIFORNIA
PyGmMy
OwL. — Taken as far north as Willow River. 57. Dryobates
villosus
leucomelas. NoRTHERN
Harry
Woop-
PECKER.— Common at Quesnelle and in the mountains. 57. Dryobates
villosus
leucomelas. NoRTHERN
Harry
Woop-
PECKER.— Common at Quesnelle and in the mountains. 58. Dryobates
villosus
hyloscopus. CABANIS’S
WOODPECKER. — Breeding at 158-Mile House. 58. Dryobates
villosus
hyloscopus. CABANIS’S
WOODPECKER. — Breeding at 158-Mile House. cae cae cae ks,
Birds
of
the
Cariboo
District,
B. 282 59. Dryobates pubescens homorus, BATCHELDER’S WoOODPECKER. —Scarce; not observed during the winter months. 59. Dryobates pubescens homorus, BATCHELDER’S WoOODPECKER. —Scarce; not observed during the winter months. 60. Picoides
arcticus. ArRcTic
THREE-TOED
WoOODPECKER.—
Not
uncommon. As far as could be judged without actual comparison, all
the specimens taken were fully as large as eastern birds. The subspecies
lately described by Mr. Outram Bangs occurs in the Okanagan district to
the southward. 60. Picoides
arcticus. ArRcTic
THREE-TOED
WoOODPECKER.—
Not
uncommon. As far as could be judged without actual comparison, all
the specimens taken were fully as large as eastern birds. The subspecies
lately described by Mr. Plate X. Outram Bangs occurs in the Okanagan district to
the southward. 61. Picoides americanus.— Taken from Willow River to Clinton;
breeds throughout this region. 62. Sphyrapicus
varius
nuchalis. RED-NAPED
SAPSUCKER.—
Common summer resident. 63. Ceophleus
pileatus. PILEATED
WoopprecKER.—
Resident;
observed as far north as Willow River. 64. Colaptes auratus. FLICKER. p
65. Colaptes cafer collaris. RED-SHAFTED FLICKER. 65. Co aptes
ca e
co a s. S
C
. In the neighborhood of the 158-Mile House both species occur and
interbreed. From one nest hole I took seven nestlings, which varied
from
typical
cafer
collars
to
nearly
typical
auratus. ; 66. Cypseloides niger. BLAcK Swirr.— Observed in the southern
portion of the district. p
67. Selasphorus
alleni. ALLEN’s
HUMMINGBIRD.—
Breeding
near
158-Mile House. 68. Stellula calliope. CaALLiopE HUMMINGBIRD.— Breeding in the
mountains west of Clinton in the Lilloet district. 69. Empidonax
traillii
alnorum. ALDER
FLYCATCHER.—
Breeding
birds taken at Quesnelle were closer to alvorum than to typical
trailliz; the latter is the species breeding in the southern portion of the
district. 70. Otocoris
alpestris
leucolema. PaLttip
HoRNED
LARK.—
Breeding above timber line near Barkerville. The spotted young are
darker in coloration than would be expected from the color of the adult. 71. Otocoris
alpestris
merrilliii
Dusky
HorNep
LArk.—
Breeding
on Chilcotin plateau. p
72. Xanthocephalus
xanthocephalus. YELLOW-HEADED
BLACKBIRD. — Noticed only at 158-Mile House, as a straggler. 73. Carpodacus cassini. CAssrIN’s PURPLE FINCH.— Summer resi-
dent at Soda Creek, and probably also at Quesnelle. 74. Leucosticte tephrocotis. GRAY-cROWNED LeEucosTicTE.— Breed-
ing above timber line near Barkerville. The young were fully fledged
the last week in July. 75. Leucosticte
tephrocotis
littoralis. HEPBURN’s
LEUCOSTICTE.—
After identifying the typical species as the species breeding in the dis-
trict, I was surprised to find /¢ttoral/s the common winter visitant around
Quesnelle, where no ¢ephrocotis were then seen. p
76. Acanthis
hornemannii
exilipes. Hoary
Rerppori.—I
took
one nearly typical example at Quesnelle, and also have several taken by
Mr. Sidney Williams at that place. Smithsonian Libraries and Archives Smithsonian Libraries and Archives This file was generated 1 April 2024 at 12:39 UTC Brooks, Allan. 1903. "Notes on the Birds of the Cariboo District, British
Columbia." The Auk 20, 277–284. https://doi.org/10.2307/4069791. View This Item Online: https://www.biodiversitylibrary.org/item/54527
DOI: https://doi.org/10.2307/4069791 Permalink: https://www.biodiversitylibrary.org/partpdf/89916 Copyright & Reuse Copyright Status: Public domain. The BHL considers that this work is no longer under
copyright protection. This document was created from content at the Biodiversity Heritage Library, the world's
largest open access digital library for biodiversity literature and archives. Visit BHL at
https://www.biodiversitylibrary.org. This file was generated 1 April 2024 at 12:39 UTC
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Skeletal Muscle Health and Cognitive Function: A Narrative Review
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Sophia X. Sui
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Citation: Sui, S.X.; Williams, L.J.;
Holloway-Kew, K.L.; Hyde, N.K.;
Pasco, J.A. Skeletal Muscle Health
and Cognitive Function: A Narrative
Review. Int. J. Mol. Sci. 2021, 22, 255. https://doi.org/10.3390/
ijms22010255 Citation: Sui, S.X.; Williams, L.J.;
Holloway-Kew, K.L.; Hyde, N.K.;
Pasco, J.A. Skeletal Muscle Health
and Cognitive Function: A Narrative
Review. Int. J. Mol. Sci. 2021, 22, 255. https://doi.org/10.3390/
ijms22010255 Keywords: skeletal muscle health; sarcopenia; cognitive function; dementia; cognitive decline;
cognitive impairment; vitamin D; inflammation; oxidative stress; lifestyle risk factors Received: 29 October 2020
Accepted: 22 December 2020
Published: 29 December 2020 Received: 29 October 2020
Accepted: 22 December 2020
Published: 29 December 2020 Received: 29 October 2020
Accepted: 22 December 2020
Published: 29 December 2020 Received: 29 October 2020
Accepted: 22 December 2020
Published: 29 December 2020 Emerging evidence indicates that sarcopenia is associated with an increased likelihood
of cognitive impairment and hence the development of dementia. This review of the
literature summarises current knowledge about potential links between sarcopenia and
dementia, risk factors and underlying biological mechanisms. Publisher’s Note: MDPI stays neu-
tral with regard to jurisdictional clai-
ms in published maps and institutio-
nal affiliations. International Journal of
Molecular Sciences International Journal of
Molecular Sciences International Journal of
Molecular Sciences International Journal of
Molecular Sciences Review
Skeletal Muscle Health and Cognitive Function:
A Narrative Review na J. Williams 1
, Kara L. Holloway-Kew 1
, Natalie K. Hyde 1
and Julie A. Pasco 1,2,3,4 Sophia X. Sui 1,*
, Lana J. Williams 1
, Kara L. Holloway-Kew 1
, Natalie K. Hyde 1
and J 1
IMPACT—The Institute for Mental and Physical Health and Clinical Translation, Deakin University,
Barwon Health, Geelong, VIC 3220, Australia; l.williams@deakin.edu.au (L.J.W.);
k.holloway@deakin.edu.au (K.L.H.-K.); natalie.hyde@deakin.edu.au (N.K.H.);
julie.pasco@deakin.edu.au (J.A.P.) 1
IMPACT—The Institute for Mental and Physical Health and Clinical Translation, Deakin University,
Barwon Health, Geelong, VIC 3220, Australia; l.williams@deakin.edu.au (L.J.W.);
k.holloway@deakin.edu.au (K.L.H.-K.); natalie.hyde@deakin.edu.au (N.K.H.);
julie.pasco@deakin.edu.au (J.A.P.) 2
Department of Medicine-Western Health, The University of Melbourne, St Albans, VIC 3021, Australia
3
Department of Epidemiology and Preventive Medicine, Monash University, Melbourne, VIC 3181, Australia
4
Barwon Health, University Hospital Geelong, Geelong, VIC 3220, Australia
*
Correspondence: ssui@deakin.edu.au or suixinad@gmail.com; Tel.: +61-3-42153306; Fax: +61-3-42153491 4
Barwon Health, University Hospital Geelong, Geelong, VIC 3220, Australia Abstract: Sarcopenia is the loss of skeletal muscle mass and function with advancing age. It involves
both complex genetic and modifiable risk factors, such as lack of exercise, malnutrition and reduced
neurological drive. Cognitive decline refers to diminished or impaired mental and/or intellectual
functioning. Contracting skeletal muscle is a major source of neurotrophic factors, including brain-
derived neurotrophic factor, which regulate synapses in the brain. Furthermore, skeletal muscle
activity has important immune and redox effects that modify brain function and reduce muscle
catabolism. The identification of common risk factors and underlying mechanisms for sarcopenia and
cognition may allow the development of targeted interventions that slow or reverse sarcopenia and
also certain forms of cognitive decline. However, the links between cognition and skeletal muscle
have not been elucidated fully. This review provides a critical appraisal of the literature on the
relationship between skeletal muscle health and cognition. The literature suggests that sarcopenia
and cognitive decline share pathophysiological pathways. Ageing plays a role in both skeletal muscle
deterioration and cognitive decline. Furthermore, lifestyle risk factors, such as physical inactivity,
poor diet and smoking, are common to both disorders, so their potential role in the muscle–brain
relationship warrants investigation. 1. Dementia Dementia is a condition that encompasses many progressive and acquired neurocog-
nitive disorders [1]. Its core feature is a loss of intellectual abilities severe enough to disturb
social and occupational functioning. Dementia affects multiple cognitive domains, such
as executive function, complex attention, learning and memory, language and perceptual
motor and social cognition [2]. Worldwide, approximately 50 million people have dementia,
and this number is projected to reach 82 million by 2030; an estimated 5–8% of people aged
60+ years have dementia [3]. Copyright: © 2020 by the authors. Li-
censee MDPI, Basel,
Switzerland. This article is an open access article
distributed under the terms and con-
ditions of the Creative Commons At-
tribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/). Alzheimer’s disease (AD) is a cause of dementia, accounting for 50–70% of dementia
cases. AD is a progressive, degenerative disorder involving loss of memory, thinking
and language skills and behavioural changes [1]. Globally, nearly 44 million people were
affected by Alzheimer’s disease in 2016 [4]. One in nine people aged 65+ years has AD, https://www.mdpi.com/journal/ijms Int. J. Mol. Sci. 2021, 22, 255. https://doi.org/10.3390/ijms22010255 Int. J. Mol. Sci. 2021, 22, 255 2 of 22 and its prevalence increases with age. In 2019, 3% of people aged 65–74 years had AD, 17%
of people aged 75–84 years and 32% of those aged 85+ years [4]. and its prevalence increases with age. In 2019, 3% of people aged 65–74 years had AD, 17%
of people aged 75–84 years and 32% of those aged 85+ years [4]. Mild cognitive impairment (MCI) is a pre-dementia stage—a condition between
normal, age-related cognitive decline and dementia [5]. Cognitive impairment, or cognitive
decline, is associated with increased mortality and substantially reduces quality of life [6–8]. The prevalence of cognitive impairment varies depending on the criteria and participants’
demographic characteristics; however, it increases with age [9,10] and appears to be higher
in men [11]. People with MCI are usually able to perform normal activities and compensate for
small changes with minimal difficulty. MCI is categorised either as an amnestic MCI
subtype in which memory is impaired or as a non-amnestic MCI subtype [6]. Most studies
measure the prevalence of MCI in the general population as 16–20% [6]. However, its
prevalence varies considerably with the criteria, definitions and assessment instruments
applied [6]. The few existing measurements of MCI incidence rates range from 5 to 168
per 1000 person-years [6]. 2. Sarcopenia Sarcopenia is the age-related deterioration of skeletal muscle and a key component
of physical frailty, including poor physical function and muscle strength. Prevalence
estimates vary from 9.9% to 40.4% due to differences in demographics across geographical
areas [14,15] and also to the use of different criteria for identifying cases. Moreover, the
application of different case definitions to the same population can cause sarcopenia
prevalence estimates to vary by 40% [14,16–21]. Data from the Geelong Osteoporosis
Study (GOS) recently described the consequences of applying different criteria and cut-
off points in a homogeneous sample to assess prevalence estimates for the Australian
population [22]. The main finding was that, across several definitions, the prevalence
of sarcopenia increased with increasing age; however, the varied criteria and cut-off
points resulted in inconsistent findings regarding case ascertainment for sarcopenia. The
varied levels of agreement between the definitions highlight the need to carefully interpret
studies that report sarcopenia prevalence in the context of the population studied and the
criteria employed. Sarcopenia components such as muscle strength and mass decline with age. The aver-
age decline from peak muscle strength by age 40 years is 16.0%, and 40.9% for people aged
60+ years [23]. Muscle mass decreases by approximately 3–8% per decade after 30 years of
age and accelerates after age 60 years [24]. There appears to be a sex difference in the rate
of decline in muscle mass and strength, with women experiencing a greater rate of decline
after menopause [25]. Information from the GOS recently reported the first normative
data for handgrip strength and muscle quality and described the relationship with age,
anthropometry and measures of body composition in women [26]. This population-based
study contributes to the evidence base by providing geographic-specific data for assessing
muscle weakness in conditions such as sarcopenia and frailty. Similar data are needed
for men. Some studies that have investigated the overlap between frailty and sarcopenia have
demonstrated that people with frailty are likely to have sarcopenia, but not all people
with sarcopenia are frail [27]. For example, a cross-sectional study in Singapore examined
participants attending an outpatient clinic where frailty was assessed using the Edmonton
Frail Scale [28] and sarcopenia was assessed using the rapid five item (SARC-F) question-
naire [29]. Of 115 patients aged 65+ years, 44.3% of patients had sarcopenia, 31 (27.0%) were
frail, and 27 (23.5%) were both frail and sarcopenic. 1. Dementia Impaired cognitive function predicts dementia in later life [6,12],
and multi-domain MCI confers a higher risk of progressing to dementia than single-
domain MCI [6]. There are currently no pharmacological or psychological therapies for
dementia [13]; however, MCI is potentially recoverable, and sufferers do not necessarily
develop dementia [2]. Intervention at the MCI stage may enable modification of the
trajectory towards dementia [2]. 3. Sarcopenia and Cognitive Function While the literature describes bi-directional associations between physical and mental
decline, little is known about skeletal muscle deficits as risk factors for poor cognitive per-
formance. Although results are equivocal, emerging evidence indicates that sarcopenia is
associated with an increased likelihood of cognitive impairment. A systematic review and
meta-analysis found that sarcopenia was associated with cognitive dysfunction (adjusted
odds ratio 2.2, 95% CI 1.2–4.2) [31]. This association was not modified by ethnicity, sex
or assessment instruments [31]. However, this meta-analysis only located cross-sectional
studies [31]; prospective studies are warranted to identify potential causal relationships. A recent cross-sectional study [32] examined the association between sarcopenia and cog-
nitive impairment in 201 older Korean women. Cognitive function was assessed using
the Mini-Mental State Examination (MMSE), while sarcopenia was identified based on the
Asian Working Group for Sarcopenia’s definition. This study reported that women with
pre-sarcopenia and sarcopenia were two and five times more likely to have cognitive im-
pairment, respectively, than women who were non-sarcopenic. In contrast, a French study
involving 3025 women aged 75+ years, which examined associations between cognitive
impairment and six operative sarcopenia definitions [33], found that sarcopenia was not
associated with cognitive impairment, regardless of the definition applied. Furthermore,
researchers in the United States of America (USA) reported that sarcopenia was not as-
sociated with cognitive functioning in adults aged 60–69 years, but it was for those aged
70+ years [34]. These contradictory results may be due to the components of sarcopenia
assessed, which might play different roles in linking physical and mental decline. Th
f
i
i ll
l
h
d
i
d
ll The components of sarcopenia, especially muscle strength and gait speed, are well
documented as being associated with cognitive function. However, how muscle mass
contributes independently of the other components of sarcopenia remains unclear. Muscle
mass is associated with muscle strength, possibly in a non-linear way [33], but the latter may
be a better predictor of cognitive decline [35]. For example, a cross-sectional study of 223 US
adults aged 40+ (mean age 68.1 years, 35% male), drawn from the general community,
examined the effect of sarcopenia on physical and cognitive functioning [36]. 2. Sarcopenia Of those with frailty, 87.1% also had Int. J. Mol. Sci. 2021, 22, 255 3 of 22 sarcopenia, whereas 47.1% of sarcopenic patients met the criteria for frailty. A study [30]
in the Netherlands included data from 227 participants (aged 65+ years) in community
care settings. Sarcopenia was identified using the algorithm from the European Working
Group on Sarcopenia in Older People 2010, while physical frailty was assessed by using
the Fried criteria and the International Association of Nutrition and Aging proposed frailty
scale. Sarcopenia was identified in 23.3% and physical frailty in 8.4–9.3% of the participants. The risk of having sarcopenia increased with age in those without frailty; however, the
older people with frailty were 60% more likely to have sarcopenia. It should be noted that
the criteria for identifying sarcopenia and frailty were applied differently; however, the
findings were consistent. 4. Muscle Mass and Cognitive Function In a cross-sectional study, Nourbashemi and colleagues (2002) [38] examined whether
lean mass was associated with cognitive deficits among 7105 women. Lean mass was
measured using dual-energy X-ray absorptiometry (DXA) and general cognitive function
using the short portable mental status questionnaire (SPMSQ). The authors found that
women in the lowest quartile of lean mass had a 1.43-times higher risk of general cognitive
impairment than those in the highest quartile of lean mass. However, the authors pointed
out that this study had several limitations, notably using SPSMQ to screen for cognitive
impairment without clinical assessment of dementia, not assessing potential confounders
(e.g., physical activity) and its single-sex sample [38]. Similarly, in a German cross-sectional
study of the relationship between cognitive function, body composition and nutrition,
involving 4095 hospitalised patients (71.3% female), cognitive function was evaluated
using MMSE and lean mass using BIA [39]. Results showed that a 5.9% loss of lean mass
was associated with an increase in score from 2.1 to 3.0, indicating cognitive deterioration. A cross-sectional study of 51 healthy and community-dwelling older men in the United
Kingdom (UK) reported conflicting results [40]. The study measured muscle volume
in the neck area [41], general cognitive function using MMSE and cognitive domains of
memory and executive function using Rey’s auditory–verbal declarative memory test
and the controlled word association test, respectively [40]. It is noteworthy that muscle
volume is rarely reported in the literature and is different from muscle mass. An estimate of
prior general cognitive ability was also assessed using Benton’s visual retention test (a test
for visual memory) and the national adult reading test. This study found no association
between neck muscle volume and cognitive abilities [40] and found that total muscle
volume was negatively associated with estimated prior cognitive ability [40]. The results
suggested that individuals with lower cognitive abilities are more likely to have larger
muscle size as they age [40]. The authors speculated that this finding may have been due
to men with lower cognitive function being more likely to engage in manual work. In a prospective study of sarcopenia as a risk factor for cognitive impairment, 297 par-
ticipants aged 65+ years without cognitive impairment at baseline were followed over a
period of five years [42]. 3. Sarcopenia and Cognitive Function It measured
muscle strength using a handheld dynamometer and lean mass (a surrogate measure of
muscle mass) using bioelectrical impedance analysis (BIA); participants exhibiting low
lean mass were classified as pre-sarcopenic, those with low lean mass and muscle strength
were considered sarcopenic, and those with high lean mass and low muscle strength were
categorised as non-sarcopenic [36]. The Montreal Cognitive Assessment was used to assess
visuospatial function, executive function, attention, language, memory and orientation;
the Ascertaining Dementia questionnaire was used to identify changes in memory and
problem-solving ability [36]. Individuals with sarcopenia were six times more likely to
have cognitive impairment than the non-sarcopenic group (healthy controls) [36]. After
adjusting for confounders, there was a threefold greater risk of cognitive impairment for
the sarcopenia group [36]. Thus, muscle strength, rather than muscle mass, appeared
to drive the relationship between sarcopenia and cognitive impairment, suggesting that
interventions designed to improve muscle strength may also reduce cognitive decline in
middle-aged and elderly people [36]. This study was limited by its cross-sectional design Int. J. Mol. Sci. 2021, 22, 255 4 of 22 and measurement of lean mass without adjusting for body mass index [36]. Data from the
GOS recently investigated components of sarcopenia individually in relation to cognitive
function [37]. The main finding was that muscle strength and gait speed, rather than
muscle mass, are better indicators of poor cognitive function, especially in the domains of
information processing (psychomotor function), visual attention and overall performance,
even after accounting for differences in age, education status and physical activity. g
g
p y
y
Further research is warranted to identify why studies have produced conflicting
conclusions, which components of sarcopenia are associated with cognitive impairment
and whether they are risk factors for cognitive decline. The following sections present a
critical appraisal of the evidence for the relationship between muscle mass and function
(components of sarcopenia) and cognitive function. 5. Muscle Strength and Cognitive Function Cross-sectional studies have consistently demonstrated an association between muscle
strength and cognitive function. A study of 3025 French women aged 75+ years, recruited
from the community, measured general cognitive function by SPMSQ and muscle strength
by handgrip strength (HGS) [33]. Lower HGS was associated with cognitive impairment
(OR 1.81, 95% CI 1.33–2.46) [33]. Similarly, in a study of Japanese adults aged 85 years
(90 men, 117 women), recruited from the community, those with higher MMSE scores had
greater mean HGS in both the right hand (21.8 ± 7.1 vs. 19.3 ± 5.8 kg, p = 0.009) and the
left (20.6 ± 6.7 vs. 17.9 ± 5.5 kg, p = 0.003) and greater isometric leg extensor strength
(22.7 ± 8.7 vs. 20.7 ± 9.1 kg, p = 0.18) [47]. This association persisted after adjustment
for confounders [47]. These two studies, however, included mainly cognitively normal
individuals, suggesting possible sample bias. Moreover, while the studies mentioned above
suggest a positive association between muscle strength and general cognitive function,
they offer no insight as to which domains of cognitive function are most closely related to
muscle strength. g
The association between muscle strength and specific cognitive domains has been
examined using neuropsychological batteries. For instance, in a study involving 1799 US
participants aged 60+ years, knee extensor isokinetic strength was measured using a kinetic
communicator isokinetic dynamometer and visual spatial and motor speed processing
using the digit symbol substitution test (DSST) [48]. The participants were divided into four
groups by quartiles of quadriceps strength. DSST scores were greater in higher quadriceps
strength groups, indicating that muscle strength is associated with speed of processing and
visual–spatial processing [48]. In a study in the Netherlands of 555 participants at ages
85 (35% men) and 89 years (29% men), the adjusted scores of cognitive tests were categorised
into tertiles; HGS was associated with scores in tests for processing speed and memory
for both age groups, but not with attention at age 89 years [49]. These studies suggest
that muscle strength is associated with certain cognitive domains and less related to other
domains in older adults. However, specific cognitive domains are described poorly in the
current literature. While cross-sectional studies support an association between muscle strength and
cognitive function in general and specific domains, some longitudinal studies indicate a
bi-directional association between poor muscle strength and poor cognitive function. 4. Muscle Mass and Cognitive Function Mean lean mass did not differ between groups (normal cognitive
function, MCI and dementia); thus, no significant association between lean mass and the
risk of developing cognitive impairment was detected. However, the authors suggested
that this lack of association may have been due to insufficient statistical power [40,42]. There may be sex differences in the association between muscle mass and cognitive
function, as muscle mass varies substantially between men and women [33]. However, no
evidence has confirmed this hypothesis. For example, in a recent longitudinal study in
Hong Kong, 2737 cognitively healthy men and women aged 65+ years were followed over
four years [43]. Lean mass was measured using DXA and general cognitive function using
MMSE. Lower lean mass was associated with a higher risk of general cognitive decline in
men; however, this association was not sustained after adjusting for confounders, and no
relationship between lean mass and general cognitive decline was found in women. The Int. J. Mol. Sci. 2021, 22, 255 5 of 22 sample was not randomly selected, which could have led to a bias towards healthier older
adults, both physically and cognitively. Intervention studies of muscle mass and changes in cognitive function have produced
unconvincing results. For example, Lauque et al., 2004 [44] conducted a randomised trial
of the influence of oral nutritional supplements on physical and mental status, including
body composition and cognitive function, in patients with AD aged 65+ years, over three
months. Forty-six patients were treated with nutritional supplements for three months and
45 received their usual care; lean mass increased in the nutrition supplement group, but no
change in cognitive function was detected. Some studies have examined body composition in patients with AD and dementia. A cross-sectional study of US adults aged 60+ years identified as cognitively normal
(n = 70) or with early-stage AD (n = 70) had body composition measured by DXA and
cognitive function by a standardised psychometric battery [45]. Lean mass was lower in the
patients with AD, after controlling for sex [45]. In contrast, in another cross-sectional study
involving 1462 women aged 75 years or older in France, in which lean mass was again
measured using DXA but cognitive function was evaluated by SPMSQ or MMSE, lean
mass was not associated with dementia [46]. These cross-sectional studies demonstrate
the limited evidence for an association between muscle mass, cognitive impairment and
dementia. 5. Muscle Strength and Cognitive Function In a
study of 2160 non-institutionalised Mexican Americans (57.5% women) aged 65+ years,
HGS at baseline was associated with greater cognitive decline (MMSE; β estimate = 1.28, Int. J. Mol. Sci. 2021, 22, 255 6 of 22 se = 0.16; p = 0.0001) over a period of six years [50]. In 2381 Mexican American men
and women without disabilities aged 65+ years, a decline in HGS was observed over
seven years for participants with poor global cognitive function (measured by MMSE)
compared to those with good cognitive function [51]. The Women’s Health Initiative
Memory Study (WHIMS) in the US, involving 1793 women aged 65–80 years, measured
reciprocal changes in general cognitive function (MMSE scores) and HGS over six years [52]. Another longitudinal study reported associations between reduced HGS decline and better
attention, memory and processing speed at baseline; however, lower baseline HGS was not
associated with an accelerated decline in the measured cognitive domains [49]. Further
longitudinal studies are required to determine if a bi-directional relationship exists. g
q
p
The relationship between muscle strength and cognitive function has been examined
in intervention studies. A randomised study in Austria assessed the effect of structured
strength training on cognitive function in 42 men and women (mean age 86.8 years) with
cognitive impairment and frailty and found that muscle strength in the muscle training
group increased over 10 weeks compared to the control group [53]. Even though a linear
relationship was observed between muscle strength and MMSE scores in the muscle train-
ing group, the mean MMSE scores of the training and control groups did not differ [53]. The short intervention period may have contributed to this finding. Other studies have
investigated whether muscle strength intervention improves cognitive function in specific
domains. A Brazilian study involved 62 older adults aged 65–75 years undertaking 24-week
resistance training at two intensities to examine the impact of muscle strength training
on cognitive function [54]. Participants were randomly assigned to control, experimen-
tal moderate- and experimental high-intensity training groups [54]. Cognitive function
was tested using the Wechsler adult intelligence scale, third edition (WAIS-III), to assess
specific cognitive domains, such as central executive, short-term memory and long-term
memory [54]. The training groups showed improvement on neuropsychological tests,
such as the forward digit span and immediate recall tests, indicating that the intervention
improved cognitive function [54]. Similarly, Berryman et al. 5. Muscle Strength and Cognitive Function studied the effect of eight
weeks of aerobic strength training on the executive function of 47 healthy adults (mean
age 70.7 ± 5.6 years) and reported increased muscle strength and improved executive
function [55]. These studies indicate that improving muscle strength improves cognitive
function, at least in some domains. While general cognitive function has been assessed using global cognitive tests, such
as MMSE and its versions, these tests have little ability to identify subtle differences
within healthy populations [56]. Further research must incorporate measures of general
and specific cognitive function to clarify the relationships between muscle strength and
cognitive function. g
A cross-sectional study involving 1038 Korean men and women with severe cognitive
impairment, aged 65+ years, reported that each 8-kg decrease in HGS was associated with
a 59% increased likelihood of dementia (adjusted OR 1.59; 95% CI 1.19–2.14) [57]. Similarly,
the Religious Orders Study, involving 877 US men and women without dementia, reported
that each 1 kg deficit in baseline HGS conferred a 1.5% greater risk of developing AD over
5.7 years (adjusted hazard ratio 0.986; 95% CI 0.973–0.998) [58]. 6. Physical Performance and Cognitive Function Gait speed has been associated with health outcomes and lifespan [59] and is consid-
ered to indicate higher-level cognitive functioning, because it is involved in complicated
cognitive functions such as attention, memory, motor function and perception. However,
most studies perceive gait performance and cognitive function as independent constructs. Several cross-sectional studies suggest that gait speed is associated with cognitive function. A study of 4000 Chinese men and women from the community examined the associ-
ation between cognitive function and physical performance, using a 6-m walk speed
test and chair stand test. These authors found that the cognitive impairment group had
poorer performance in gait speed tests than the non-cognitively impaired control group Int. J. Mol. Sci. 2021, 22, 255 7 of 22 (0.89 ± 0.024 vs. 1.02 ± 0.004 m/s in men and 0.85 ± 0.009 vs. 0.93 ± 0.005 m/s in
women, both p < 0.001) and chair stand tests (13.99 ± 0.05 s vs. 12.57 ± 0.09 s in men and
14.45 ± 0.27 s vs. 13.07 ± 0.12 s in women, both p < 0.001) [60]. Additionally, a US study
involving 44 men and women with amnestic MCI (mean age = 79.3 ± 4.7 years), 62 with
non-amnestic MCI (mean age 81.8 ± 6.2 years) and 295 healthy individuals (mean age
81.8 ± 6.2 years) compared gait performance across the groups [61]. Tests of gait perfor-
mance were conducted using computer-based analyses of gait ability that included pace,
rhythm and variability. Cognitive function was assessed using a neuropsychological test
battery for general cognitive function and specific cognitive domains, including memory,
executive function, attention and language [61]. The results showed that gait, even if
measured in different ways, was worse in participants with MCI than in the controls. Gait speed has been associated with specific cognitive domains, such as executive
function, visuospatial ability and psychomotor function [62], and poor gait performance
with executive function impairment. For example, Coppin et al., 2006 [63] reported slower
gait speed in participants with poor executive function than those with high executive
function. Several specific cognitive domains were not associated with gait in other studies. For instance, Martin et al. [62] reported that gait measures were not associated with memory,
perhaps due to test sensitivity or lack of study power, but possibly because gait is only
associated with specific domains of cognitive function [64]. 7. Muscle Quality, Muscle Density and Cognitive Function Muscle quality is defined as the force generated by each volumetric unit of muscle
tissue and may reflect the amount of contractile protein, fat infiltration (myosteatosis),
aerobic capacity and other physiological properties of the muscle [74,75]. However, as there
is no standardised protocol for quantifying muscle quality, few studies have examined
low muscle quality in association with cognitive function. One US study examined the
relationship between muscle quality (isokinetic strength in relation to leg muscle mass)
and cognitive function in adults aged 60+ years, finding a positive relationship in both
sexes [76]. Bone mineral density and lean mass are correlated [77]. A UK study investigated
cognitive impairment and bone using peripheral quantitative computed tomography
(pQCT), and reported that gait speed, but not bone density/structure, was associated with
MCI [78]. A similar Italian study suggested that low bone mineral density is an early
marker of cognitive decline in the elderly [79]. These studies used MMSE to screen for
cognitive impairment, which examines global cognitive function. Data from the GOS [80] recently examined pQCT-derived muscle density (a surrogate
measure of muscle quality) and specific domains of cognitive function. This study included
281 men (aged 60–95 years) whose radial and tibial muscle density were measured using
pQCT and body fat and appendicular lean mass were measured using DXA. Cognitive
function was assessed in different domains using CogState Brief Battery. Regression
analyses revealed that muscle density was associated with psychomotor function and
visual learning, independent of education and physical activity. While there was some
evidence that serum Tumour Necrosis Factor alpha (TNF-α) attenuated the association
between radial muscle density and psychomotor function, there was little other evidence
that inflammation and adiposity explained the association between muscle density and
cognitive function, at least with the parameters investigated. The age-related associations
did not occur simultaneously between muscle density and all cognitive domains that were
tested. 8. Potential Mechanisms
8.1. Vitamin D Circulating vitamin D helps to sustain both skeletal muscle and brain function. A
systematic literature review and meta-analysis of 13 randomised controlled studies of the
effect of supplemental vitamin D on muscle strength, gait and balance in older adults
concluded that it improves muscle strength and balance [81]. There is likely to be no ideal
level for supplemental vitamin D, which will depend on the background vitamin D status,
from dietary sources and exposure to ultraviolet radiation (UV), as a threshold effect is
likely. Vitamin D deficiency also plays a role in brain function deficits. Another systematic
literature review and meta-analysis (37 studies) found that low vitamin D is associated
with poor cognitive function and a higher risk of developing dementia [82]. More research
is needed to determine if low vitamin D concurrently influences both sarcopenia and
cognitive decline. 6. Physical Performance and Cognitive Function Some longitudinal studies show that reduced gait speed in ageing predicts declines in
general cognitive function and several specific domains. A US study of 204 healthy older
adults (58% female) found that gait speed declined by 0.02 m/s/year for up to 12 years
prior to the onset of MCI, as assessed using standardised neurologic examinations [65]. Similarly, in the Health, Ageing and Body Composition (Health ABC) Study, involving
2776 men and women aged 75–85 years, DSST scores underwent the largest declines in
participants in the lowest quartile of gait speed, indicating that gait speed predicts decline
in attention and psychomotor speed in the elderly [66]. Conversely, several studies have demonstrated that general and specific cognitive
decline predict gait speed decline. In the Health ABC Study, assessment of 2349 men and
women (mean age 75.6 years) over three years showed that lower global cognitive function
and executive function were associated with greater gait speed decline [67]. Executive
cognitive deficits during ageing were found to account for gait slowing [68]. Lower general
cognitive function, verbal memory and executive function were associated with greater
gait speed decline each year [69]. More specifically, Soumare et al., 2009 reported that
poorer verbal fluency and slower psychomotor speed were associated with larger declines
in gait speed [70]. Several studies have investigated whether fast gait speed predicts cognitive decline. In an Italian longitudinal and population-based study involving 660 older adults aged
65+ years, followed over three years, usual gait speed, fast speed and speed during
“walking while talking” were measured at baseline and follow-up; cognitive function
was assessed using MMSE at baseline and follow-up and adjusted for confounders [71]. Only fast gait speed predicted general cognitive decline [71]. Several other studies have
examined bi-directional associations between muscle function and cognitive function. For
example, change in general cognitive function was associated with change in physical
performance, including gait speed; however, baseline physical performance, including gait
speed, was not associated with cognitive change [52]. A 2004 systematic review included seven studies comparing gait in dementia and
healthy older adults and reported that the former had shortened step length and lower
walking speed [72]. A more recent review of the association between gait and cognitive
function in epidemiological and neuropsychological studies of gait disorders and dementia
revealed that gait disorders were associated with moderate and severe dementia, but not
mild dementia [73]. Int. J. Mol. Sci. 6. Physical Performance and Cognitive Function 2021, 22, 255 8 of 22 8.2. Inflammation and Oxidative Stress Fat infiltration into skeletal muscle occurs with ageing and is a characteristic of
sarcopenic obesity, whereby sarcopenia exists in the face of excessive body fat accumulation. As obesity is an inflammatory state [83,84] and inflammation has detrimental effects on
both muscle [85] and brain [76], it is plausible that inflammation, or indeed adiposity itself,
might play a role in linking low muscle density with poor cognitive function. Systemic, chronic, low-grade inflammation in ageing, termed inflamm-ageing [86], is a
major risk factor for both mental and physical disease [87,88]. Inflamm-ageing contributes
to changes in skeletal muscle properties [89] and sarcopenia [85] and mediates AD and
cognitive deficits in the elderly [85,90]. Biomarkers of inflammation, such as interleukin 6 Int. J. Mol. Sci. 2021, 22, 255 9 of 22 (IL-6) and TNF-α, have been associated with muscle atrophy in both human and animal
studies [85,91,92] and age-related cognitive decline in humans [90,93–95]. (IL-6) and TNF-α, have been associated with muscle atrophy in both human and animal
studies [85,91,92] and age-related cognitive decline in humans [90,93–95]. Considerable evidence demonstrates a strong association between inflammation and
risk of cognitive decline. For example, a cross-sectional US study involving 269 participants
(mean age 67 years) found that IL-6 was negatively associated with MMSE [96]. Cross-
sectional studies in Germany (n = 369) [97] and Italy (n = 744) [98], involving participants
aged 65+ years, found negative associations between inflammatory markers (C-reactive
protein, CRP and IL-6) and performance in specific cognitive domains. The association
between inflammation and cognitive function has been confirmed through longitudinal
studies. In the Health ABC study of black and white men and women aged 70–79 years
over two years (n = 3013) and eight years (n = 2509) [99], general cognitive function was
measured using a modified MMSE. Individuals with higher levels of IL-6 and CRP had a
24% increased risk of cognitive decline at two-year follow-up and similar results at eight
years [99,100]. Other longitudinal studies in the Netherlands found that CRP and IL-6
were negatively associated with memory [101,102], learning [102], attention [101], cognitive
speed [101] and language [102]. A meta-analysis that included 1098 patients with AD and
1094 controls in 27 studies reported oxidative damage in peripheral blood vessels during
early-stage AD [103]. A US cross-sectional study of the pathophysiological links between sarcopenia and
dementia involved 445 women and 442 men aged 60+ years [76]. 8.3. Vitamin D, Exercise and Inflammation Vitamin D inhibits inflammation through regulating the production of inflammatory
cytokines and inhibiting the proliferation of proinflammatory cells [104], which could be
beneficial to skeletal muscle heath and brain health [105]. A Iranian study found that
vitamin D3 supplementation reduced the mRNA expression levels of IL-17A and IL-6, but
increased the level of IL-10 [106]. This study focused on patients with multiple sclerosis. Exercise through muscle contractions induces myokines (e.g., IL-6, brain-derived
neurotrophic factor, BDNF). Recovery from IL-6 peak following exercise can dampen
inflammation and oxidative burst activity; the anti-inflammatory effect of BDNF may
contribute to this recovery. It is well known that chronic exercise downregulates systemic
inflammation and is an effective management strategy for insulin resistance [107]. Thus,
exercise-induced downregulation of IL-6 and the anti-inflammatory effect of BDNF may
be related pathways. Thus, dietary, UV-associated or supplementary vitamin D combined
with exercise may affect both brain and skeletal muscle health. For example, a study in the
USA suggested that supplemental vitamin D combined with intense exercise may enhance
the recovery of muscle strength in adults who suffer a muscle damage event [108]. 9. Common Lifestyle Risk Factors Lifestyle factors are likely to mediate the association between sarcopenia and cognitive
impairment. Physical inactivity, poor diet and smoking are examples of poor health
behaviours common to both disorders. 8.2. Inflammation and Oxidative Stress Muscle quality was
assessed using isokinetic strength per unit muscle mass, and cognitive function using the
WAIS-III digit symbol [76]. This study found that high-sensitivity CRP was associated with
both cognitive impairment and poor muscle quality in women [76]. 9.1. Physical Inactivity A 2014 systematic review of 47 cohort studies highlighted that physical activity was
associated with various cognitive outcomes [109]. Over 87% of the studies and 100% of
the cross-sectional studies demonstrated a relationship between physical inactivity and
cognitive impairment, including AD [109]. Resistance training is a recognised means of
improving skeletal muscle mass and function [110]. Contracting skeletal muscles produce
cytokines [111], which affect lipid and glucose metabolism. Int. J. Mol. Sci. 2021, 22, 255 10 of 22 10 of 22 9.3. Smoking Tobacco smoke generates free radicals, causing lipid peroxidation, oxidation of protein
and other tissue damage in smokers [121]. Nicotine may influence the link between
sarcopenia and cognitive impairment, but other toxic components of cigarettes cannot
be ignored. A meta-analysis of smoking as an independent risk factor for sarcopenia included
22,515 participants across 12 studies [122]. A fixed effect model was used to estimate
sarcopenia risk for men (OR 1.12, 95% CI 1.03–1.21) and a random effect model for women
(OR 1.21 95% CI 0.92–1.59). These sex-specific results are difficult to compare because of the
different methods employed [122]. Another study found that lifelong cigarette smoking
was associated with higher prevalence of sarcopenia in older adults [123]. Reviews of epidemiological and clinical studies provide inconsistent evidence as
to whether smoking is a risk factor for cognitive impairment. Many epidemiological
studies have found that smoking is negatively associated with AD [124]. An early study
found that patients with AD and/or Parkinson’s disease are more likely to be lifelong non-
smokers than those without. However, other studies have detected a positive independent
association between smoking and AD. Note that a survivor effect may exist in large-scale
longitudinal studies: non-smokers may be more likely to suffer from AD simply due to
surviving to an age when it is more common. 9.2. Poor Diet Adequate nutrition is essential for maintaining skeletal muscle, and sarcopenia is
associated with inadequate caloric and protein intakes. Prospective studies describe the
protective effect of dietary protein for sarcopenia [112] and positive associations between
protein intake and muscle mass [113]. Furthermore, muscle mass losses are considerably
reduced in older individuals with high protein intake [114]. Malnutrition is substantially
associated with frailty and sarcopenia in hospitalised older patients [115] and in geriatric
rehabilitation patients [116]. It is well documented that poor diet promotes cognitive
decline, including AD [117,118]. Consumption of fish (a source of long-chain omega-3
fatty acids) slows cognitive decline in older people without AD, but this study does not
suggest that it can treat dementia [119]. A US longitudinal study [120] examined the
relationship between poor diet and cognitive decline in participants with a mean age of
48 years. Diet quality was assessed using a self-reported questionnaire, and cognitive
function with a neurocognitive test battery. Poorer diet was associated with reduced
attention and cognitive flexibility, visuospatial ability and perceptual speed. The lean mass was lower
in the patients with
Alzheimer’s disease,
after controlling for sex 9.4. Alcohol Consumption Alcohol consumption is associated with body composition changes, including muscle
autophagy, as ethanol inhibits protein synthesis in muscles [125]. However, the current
evidence is controversial. A meta-analysis that included 214 research articles involving
a total of 13,155 participants found no evidence to support alcohol intake as a risk factor
for sarcopenia [126]. However, a cross-sectional study conducted by Daskalopoulou et al.,
2020 [127] that investigated the risk factors of sarcopenia and sarcopenic obesity in low- and
middle-income countries reported that people who consume moderate levels of alcohol
(1–14 units per week for women and 1–21 units per week for men) were more likely to
have sarcopenic obesity (OR 1.76, 95% CI 1.21–2.57), but not sarcopenia, compared with
people who were in the no drinking or heavy drinking group. Prenatal alcohol exposure is a risk factor for children’s cognitive development [128]. In
contrast,
a systematic literature review examined 27 cohort studies (published
2007–2018) [129]. Interestingly, this review reported that moderate alcohol consump-
tion was a protective factor for better cognition in women but no difference was found in
men. Data from the GOS found that high alcohol consumption (>20 g/day) was associated
with greater tibial muscle density but not with any domains of cognitive function. Higher Int. J. Mol. Sci. 2021, 22, 255 11 of 22 11 of 22 levels of alcohol consumption did not contribute to or explain the relationships between
muscle density and cognitive function [80]. levels of alcohol consumption did not contribute to or explain the relationships between
muscle density and cognitive function [80]. 10. Summary and Conclusions The main body of text in this review included and critically examined 30 key research
articles in terms of cross-sectional studies, longitudinal studies and clinical trials that
demonstrated relationships between skeletal muscle and cognitive function (Tables 1–3). Current knowledge about the relationship suffers from a lack of data for cognitive function
in specific domains. Research to date has almost exclusively assessed associations between
overall cognition and skeletal muscle parameters. Previous studies have shown that
different methodologies for assessing cognitive performance may contribute to equivocal
results; future studies may investigate biomarkers and utilise neuroimaging techniques for
screening MCI. Table 1. Summary of studies that investigated association between skeletal muscle mass (or lean mass) and cognitive function. Table 1. Summary of studies that investigated association between skeletal muscle mass (or lean mass) and cognitive function. Author, Year;
Country/Region;
Follow-Up Period
Participant
Characteristics
Muscle Mass
Measurements
Cognitive Function
Measurements
Results
Cross-Sectional Studies
1. Nourbashemi
et al., 2002 [38];
France
7105
community-dwelling
women aged
75+ years
DXA (lean mass)
SPMSQ (focus on
orientation, memory; using
to identify cognitive
impairment in this study)
Women in the lowest
quartile of lean mass had
a 1.43-times higher risk
of general cognitive
impairment compared
with those in the highest
quartile of lean mass
2. Wirth et al., 2011
[39]; Germany
4095 (71.3% female);
hospitalised patients
BIA (lean mass)
MMSE (general cognition)
5.9% loss of lean mass
was associated with an
increased score from 2.1
to 3.0, indicating
cognitive deterioration
3. Kilgour et al.,
2013 [40]; UK
51
community-dwelling
older men mean aged
73.8 ± 1.5 years
CT (muscle volume)
MMSE (global cognition);
Rey’s auditory–verbal
declarative memory test
(memory);
the controlled word
association test (executive
function);
Benton’s visual retention
test; the national adult
reading test
No association between
neck muscle volume and
cognitive abilities; the
total muscle volume was
negatively associated
with estimated prior
cognitive ability;
individuals with lower
cognitive abilities were
more likely to have larger
muscle size as they aged
4. Burns et al.,
2010 [45]; USA
Cognitively normal
(n = 70) or with
early-stage
Alzheimer’s disease
(n = 70);
aged 60+ years
DXA (lean mass)
A standardised
psychometric battery
(Logical Memory, Free and
Cued Selective Reminding
Task, Boston Naming,
Verbal Fluency, Digit Span
Forward and Backward,
Letter–Number
Sequencing, Stroop
Color-Word Test and Block
Design MMSE
(global cognition)
The lean mass was lower
in the patients with
Alzheimer’s disease,
after controlling for sex
5. No association between
neck muscle volume and
cognitive abilities; the
total muscle volume was
negatively associated
with estimated prior
cognitive ability;
individuals with lower
cognitive abilities were
more likely to have larger
muscle size as they aged Lean mass was not
associated with dementia Mean lean mass was not
different between groups
(normal cognitive
function, mild cognitive
impairment and
dementia); thus, no
significant associations
between lean mass and
the risk of developing
cognitive impairment
were detected 10. Summary and Conclusions Author, Year;
Country/Region;
Follow-Up Period
Participant
Characteristics
Muscle Mass
Measurements
Cognitive Function
Measurements
Results
6. Sui et al., 2020
[37]; Australia
292 men aged
60+ years;
population based
DXA (lean mass)
CogState Brief
Battery (psychomotor
function, visual
identification/attention,
visual learning and
working memory
No association was
detected between lean
mass and
cognitive function
7. Sui et al.,2020
[80]; Australia
281 men aged
60+ years;
population based
pQCT (muscle
density)
CogState Brief
Battery (psychomotor
function, visual
identification/attention,
visual learning and
working memory
Muscle density was
associated with cognitive
function in the
psychomotor function
and visual learning
Longitudinal Studies Table 1. Cont. 292 men aged
60+ years;
population based 281 men aged
60+ years;
population based 8. Moon et al., 2016
[42]; South Korean; 5
years follow-up
297 community-
dwelling men and
women without
cognitive impairment
at baseline; aged
65+ years
DXA (lean mass)
Korean version of the
Consortium to Establish a
Registry for Alzheimer’s
Disease Clinical
Assessment Battery;
Korean version of the Mini
International
Neuropsychiatric
Interview; International
Working Group on MCI;
DSM-IV;
Final diagnosis of MCI,
dementia was determined
by a panel of research
neuropsychiatrists
M
d
f were detected Auyeung et al.,
11 [43]; Hong Kong;
ur years follow-up
2737 cognitively
healthy men and
women from the
community; aged
65+ years
DXA (lean mass)
MMSE (general cognition)
Lower lean mass was
associated with a highe
risk of general cognitiv
decline in men; howeve
this association was no
sustained after adjustin
for confounders and no
relationship between lea
mass and general
cognitive decline was
found in women 9. Auyeung et al.,
2011 [43]; Hong Kong;
four years follow-up
2737 cognitively
healthy men and
women from the
community; aged
65+ years
DXA (lean mass)
MMSE (general cognition)
g
risk of general cogni
decline in men; howe
this association was
sustained after adjus
for confounders and
relationship between
mass and general
cognitive decline w 2737 cognitively
healthy men and
women from the
community; aged
65+ years 9. Auyeung et al.,
2011 [43]; Hong Kong;
four years follow-up Intervention Studies 10. Cassilhas et al.,
2007 [54];
Brazil; 24 weeks
62 older adults aged
from 65 to 75 years. Lower lean mass was
associated with a higher
risk of general cognitive
decline in men; however,
this association was not
sustained after adjusting
for confounders and no
relationship between lean
mass and general
cognitive decline was
found in women 10. Summary and Conclusions Abellan van Kan
et al., 2012 [46];
France
1462
community-dwelling
women aged
75+ years
DXA (lean mass)
SPMSQ or
MMSE (general cognition)
Lean mass was not
associated with dementia No association between
neck muscle volume and
cognitive abilities; the
total muscle volume was
negatively associated
with estimated prior
cognitive ability;
individuals with lower
cognitive abilities were
more likely to have larger
muscle size as they aged Cognitively normal
(n = 70) or with
early-stage
Alzheimer’s disease
(n = 70);
aged 60+ years 12 of 22 Int. J. Mol. Sci. 2021, 22, 255 Table 1. Cont. Author, Year;
Country/Region;
Follow-Up Period
Participant
Characteristics
Muscle Mass
Measurements
Cognitive Function
Measurements
Results
6. Sui et al., 2020
[37]; Australia
292 men aged
60+ years;
population based
DXA (lean mass)
CogState Brief
Battery (psychomotor
function, visual
identification/attention,
visual learning and
working memory
No association was
detected between lean
mass and
cognitive function
7. Sui et al.,2020
[80]; Australia
281 men aged
60+ years;
population based
pQCT (muscle
density)
CogState Brief
Battery (psychomotor
function, visual
identification/attention,
visual learning and
working memory
Muscle density was
associated with cognitive
function in the
psychomotor function
and visual learning
Longitudinal Studies
8. Moon et al., 2016
[42]; South Korean; 5
years follow-up
297 community-
dwelling men and
women without
cognitive impairment
at baseline; aged
65+ years
DXA (lean mass)
Korean version of the
Consortium to Establish a
Registry for Alzheimer’s
Disease Clinical
Assessment Battery;
Korean version of the Mini
International
Neuropsychiatric
Interview; International
Working Group on MCI;
DSM-IV;
Final diagnosis of MCI,
dementia was determined
by a panel of research
neuropsychiatrists
Mean lean mass was not
different between groups
(normal cognitive
function, mild cognitive
impairment and
dementia); thus, no
significant associations
between lean mass and
the risk of developing
cognitive impairment
were detected
9. Auyeung et al.,
2011 [43]; Hong Kong;
four years follow-up
2737 cognitively
healthy men and
women from the
community; aged
65+ years
DXA (lean mass)
MMSE (general cognition)
Lower lean mass was
associated with a higher
risk of general cognitive
decline in men; however,
this association was not
sustained after adjusting
for confounders and no
relationship between lean
mass and general
cognitive decline was
found in women
Intervention Studies
62 older adults aged
from 65 to 75 years. Participants were
randomly assigned to
three groups control
WAIS III (central executive
and short term memor )
The training groups
reported impro ement in Table 1. Cont. 10. Summary and Conclusions Participants were
randomly assigned to
three groups: control,
experimental
moderate- and
experimental
high-intensity
training (six exercises
including chest press,
leg press, vertical
traction, abdominal
crunch, leg curl and
lower back)
Whole-body
plethysmograph
(lean mass) WAIS III (central executive
and short-term memory);
WSM-R (visual modality of
short-term memory);
Toulouse–Pieron’s
concentration attention test
(attention); Rey–Osterrieth
complex figure (long-term
episodic memory) The training groups
reported improvement in
neuropsychological tests,
such as the forward digit
span and immediate
recall tests, indicating
that the
intervention improved
cognitive function Whole-body
plethysmography
(lean mass) 13 of 22 Int. J. Mol. Sci. 2021, 22, 255 Table 1. Cont. Table 1. Cont. Author, Year;
Country/Region;
Follow-Up Period
Participant
Characteristics
Muscle Mass
Measurements
Cognitive Function
Measurements
Results
11. Lauque et al.,
2004 [44]; France;
3 months
Patients with
Alzheimer’s disease
aged ≥65 years;
46 patients were
treated with
nutritional
supplements for three
months and
45 received their
usual care as a
control group
DXA (lean mass)
MMSE (general cognition)
Lean mass increased in
the nutrition supplement
group; however, no
change in cognitive
function was detected
BIA: bioelectrical impedance analysis; DSM-IV: Diagnostic and Statistical Manual of Mental Disorders, fourth edition; DXA: dual-energy
X-ray absorptiometry; MMSE: Mini-Mental State Examination; pQCT: peripheral quantitative computed tomography; SPMSQ: short
portable mental status questionnaire. Table 2. Summary of studies that investigated association between skeletal muscle strength and cognitive function. Author, Year;
Country/Region;
Follow-Up Period
Participant
Characteristics
Muscle Strength
Measurements
Cognitive Function
Measurements
Results
Cross-Sectional Studies
1. Abellan van
Kan et al., 2012 [33];
France
3025 community-
dwelling women
aged 75+ years
HGS dynamometry
SPMSQ (used to
identify cognitive
impairment)
Lower HGS was associated with
cognitive impairment
2. Takata et al.,
2008 [47]; Japan
Community-
dwelling participants
aged 85 years (90
men, 117 women)
HGS dynamometry
MMSE (global
cognition)
Those with higher MMSE scores
were more likely to have greater
HGS for both the right hand
(21.8 ± 7.1 vs. 19.3 ± 5.8 kg,
p = 0.009) and the left hand
(20.6 ± 6.7 vs. 17.9 ± 5.5 kg,
p = 0.003) and greater isometric leg
extensor strength (22.7 ± 8.7 vs. 20.7 ± 9.1 kg, p = 0.18). This
association persisted after
adjustment for confounders
3. 10. Summary and Conclusions Chen et al.,
2015 [48]; USA
1799 population-
based men and
women aged
60+ years
Isokinetic strength
dynamometry
DSST (measuring the
visuospatial and
motor speed of
processing,
represented a
sensitive measure of
frontal lobe executive
functions)
The DSST scores were greater in
higher quadriceps strength groups,
indicating that muscle strength
was associated with speed of
processing and
visual–spatial processing
4. Shin et al.,
2012 [57]; Korea
1038 men and women
aged 65+ years from
Sit-to-stand score;
HGS dynamometry
Dementia (identified
using the Korean
version of GMS
Each 8-kg decrease in HGS was
associated with a 59% increased
likelihood of dementia (adjusted Table 2. Summary of studies that investigated association between skeletal muscle strength and cognitive function. Author, Year;
Country/Region;
Follow-Up Period
Participant
Characteristics
Muscle Strength
Measurements
Cognitive Function
Measurements
Results
Cross-Sectional Studies
1. Abellan van
Kan et al., 2012 [33];
France
3025 community-
dwelling women
aged 75+ years
HGS dynamometry
SPMSQ (used to
identify cognitive
impairment)
Lower HGS was associated with
cognitive impairment
2. Takata et al.,
2008 [47]; Japan
Community-
dwelling participants
aged 85 years (90
men, 117 women)
HGS dynamometry
MMSE (global
cognition)
Those with higher MMSE scores
were more likely to have greater
HGS for both the right hand
(21.8 ± 7.1 vs. 19.3 ± 5.8 kg,
p = 0.009) and the left hand
(20.6 ± 6.7 vs. 17.9 ± 5.5 kg,
p = 0.003) and greater isometric leg
extensor strength (22.7 ± 8.7 vs. 20.7 ± 9.1 kg, p = 0.18). This
association persisted after
adjustment for confounders
3. Chen et al.,
2015 [48]; USA
1799 population-
based men and
women aged
60+ years
Isokinetic strength
dynamometry
DSST (measuring the
visuospatial and
motor speed of
processing,
represented a
sensitive measure of
frontal lobe executive
functions)
The DSST scores were greater in
higher quadriceps strength groups,
indicating that muscle strength
was associated with speed of
processing and
visual–spatial processing
4. Shin et al.,
2012 [57]; Korea
1038 men and women
aged 65+ years from
the community
Sit-to-stand score;
HGS dynamometry
Dementia (identified
using the Korean
version of GMS
B3-K; CSID-K;
CERAD)
Each 8-kg decrease in HGS was
associated with a 59% increased
likelihood of dementia (adjusted
OR 1.59; 95% CI 1.19–2.14) Each 8-kg decrease in HGS was
associated with a 59% increased
likelihood of dementia (adjusted
OR 1.59; 95% CI 1.19–2.14) 14 of 22 Int. J. Mol. Sci. 2021, 22, 255 Table 2. Cont. Table 2. Cont. 10. Summary and Conclusions Table 2. Cont. Author, Year;
Country/Region;
Follow-Up Period
Participant
Characteristics
Muscle Strength
Measurements
Cognitive Function
Measurements
Results
5. Sui et al.,
2020 [37]; Australia
292 men aged
60+ years; population
based
HGS dynamometry
CogState Brief Battery
(psychomotor
function, visual iden-
tification/attention,
visual learning and
working memory
for every 1 kg increase in handgrip
strength, scores for psychomotor
function were 0.003 (log10
milliseconds) lower and for overall
cognitive function 0.02 (unitless)
higher (both indicating better
function). Longitudinal Studies
6. Taekema et al.,
2012 [49];
Netherlands
555 population-based
participants at all
ages, 85 years at base
line (35% men) and 89
(29% men) years at
follow-up
HGS dynamometry
Neuropsychological
test battery (for
assessing global
cognitive
performance,
attention, processing
speed and memory)
HGS was associated with scores in
tests for processing speed and
memory for both age groups, but
was not associated with attention
at age 89 years
7. Buchman et al.,
2007 [58]; USA;
5 years follow-up
877 men and women
without dementia
HGS dynamometry
Dementia (Mini
Mental State
Examination; Health
Interview Survey)
Each 1-kg deficit in baseline HGS
conferred a 1.5% greater risk of
developing AD over 5.7 years
(adjusted hazard ratio 0.986; 95%
CI 0.973–0.998)
8. Alfaro-Acha
et al., 2006 [50];
USA; 6 years
follow-up
2160
non-institutionalised
Mexican
Americans (57.5%
women) aged
65+ years
HGS dynamometry
MMSE (measuring
cognitive decline)
HGS at baseline was associated
with greater cognitive decline, as
assessed by MMSE
(β estimate = 1.28,
se = 0.16; p = 0.0001) over a
period of six years
9. Raji et al., 2005
[51]; USA; 7 years
follow-up
2381 Mexican
American men and
women aged
65+ years, without
disabilities
HGS dynamometry
MMSE (measuring
cognitive decline)
A decline in HGS was observed
over a period of seven years for
participants with poor global
cognitive function (measured by
MMSE) compared with those with
good cognitive function
10. Atkinson et al.,
2010 [52]; USA;
6 years follow-up
1793 women aged
65–80 years
HGS dynamometry
MMSE (measuring
cognitive decline)
Reciprocal changes in general
cognitive function (MMSE scores)
and HGS over a follow-up period
of 6 years
Intervention Studies
11. 10. Summary and Conclusions Dorner et al.,
2007 [53]; Austria;
10-week trial
42 long-term care
facility residents
(men and women,
mean age of 86.8
years) with cognitive
impairment and
frailty; intervention
through involved a
structured strength
and balance training
Increased muscle
Strength
MMSE (measuring
cognitive decline)
Muscle strength in the muscle
training group increased
compared with the control group
over a period of ten weeks [53]. Even though a linear relationship
was observed between increasing
muscle strength and improved
MMSE scores in the muscle
training group, a difference was
not detected in mean MMSE
scores between the training and Author, Year;
Country/Region;
Follow-Up Period
Participant
Characteristics
Muscle Strength
Measurements
Cognitive Function
Measurements Longitudinal Studies Reciprocal changes in general
cognitive function (MMSE scores)
and HGS over a follow-up period
of 6 years Muscle strength in the muscle
training group increased
compared with the control group
over a period of ten weeks [53]. Even though a linear relationship
was observed between increasing
muscle strength and improved
MMSE scores in the muscle
training group, a difference was
not detected in mean MMSE
scores between the training and
control groups 15 of 22 Int. J. Mol. Sci. 2021, 22, 255 Table 2. Cont. Table 2. Cont. Author, Year;
Country/Region;
Follow-Up Period
Participant
Characteristics
Muscle Strength
Measurements
Cognitive Function
Measurements
Results
12. Cassilhas et al.,
2007 [54]; Brazil;
24 weeks
62 older adults aged
from 65 to 75 years. Participants were
randomly assigned to
three groups: control,
experimental
moderate- and
experimental
high-intensity
training (six exercises
including chest press,
leg press, vertical
traction, abdominal
crunch, leg curl and
lower back)
1 RM test
WAIS III (central
executive and
short-term memory);
WSM-R (visual
modality of
short-term memory);
Toulouse–Pieron’s
concentration
attention test
(attention);
Rey–Osterrieth
complex figure
(long-term episodic
memory)
The training groups reported
improvement in
neuropsychological tests, such as
the forward digit span and
immediate recall tests, indicating
that the intervention improved
cognitive function
13. The training groups reported
improvement in
neuropsychological tests, such as
the forward digit span and
immediate recall tests, indicating
that the intervention improved
cognitive function For every 1 m/s increase
in gait speed, scores for
psychomotor function
were 0.12 lower, attention
0.08 lower and overall
cognitive function 0.49
higher (all better
function) Reported slower gait
speed in participants
with poor executive
function than those with
high executive function Gait was worse in
participants with MCI
than in the controls 10. Summary and Conclusions Berryman
et al., 2014 [55];
eight weeks
47 healthy older
adults (mean age
70.7 ± 5.6 years);
compared the effects
of three interventions:
strength training
Isokinetic strength
dynamometer
Generation cognition
(MMSE); executive
functions, memory,
processing speed
Intervention increased muscle
strength and improved executive
function
CERAD: Consortium to Establish a Registry of Alzheimer’s Disease; CSID-K: Community Screening Interview for Dementia; DSM-IV:
Diagnostic and Statistical Manual of Mental Disorders, fourth edition; DSST: digit symbol substitution test; GMS B3-K: Korean version of
Geriatric Mental State Schedule B3; HGS: handgrip strength; MMSE: Mini-Mental State Examination; SPMSQ: short portable mental status
questionnaire; WSM-R: Wechsler Adult Intelligence Scale III (WAIS III); Wechsler Memory Scale-Revised. Summary of studies that investigated associations between physical performance and cognitive function. Table 3. Summary of studies that investigated associations between physical performance a Cross-Sectional Studies 6-m walk speed
test and chair
stand test 16 of 22 Int. J. Mol. Sci. 2021, 22, 255 16 of 22 Table 3. Cont. Table 3. Cont. Author, Year,
Country/Region,
STUDY Type
Participant
Characteristics
Physical
Performance
Measurements
Cognitive Function
Measurements
Results
2. Verghese
et al., 2008 [61];
USA
44 men and women
with amnestic MCI
(mean age = 79.3 ± 4.7
years), 62 with
non-amnestic MCI
(mean age 81.8 ± 6.2
years) and 295 healthy
individuals (mean age
81.8 ± 6.2 years)
Computer-
based analyses
of gait ability
that included
pace, rhythm
and variability
Blessed Information-Memory-
Concentration test (General
cognition); FCSRT (verbal
memory); DSST, TMT-B, LFT
(executive function); TMT-A and
Digit Span
forwards-attention); Boston
Naming Test (language)
Gait was worse in
participants with MCI
than in the controls
3. Coppin et al.,
2006 [63]; USA
37
community-dwelling
individuals aged
65+ years
Complex
walking tasks;
reference
walking tasks
TMT (executive function); MMSE
(general cognition)
Reported slower gait
speed in participants
with poor executive
function than those with
high executive function
4. Martin et al.,
2013 [62];
Australia
422 older people aged
60–85 years
GAITRite
walkway
COWAT, Category Fluency,
Victoria Stroop test, WAIS-III
(Executive function/attention);
WAIS-III (Processing speed); Rey
Complex Figure copy task
(Visuospatial ability); Hopkins
Verbal Learning Test—revised,
generating scores for total
immediate recall, delayed
recall and recognition memory
and a delayed
reproduction after 20 min of the
Rey Complex Figure (memory)
Gait measures were not
associated with memory
5. Gait measures were not
associated with memory 10. Summary and Conclusions Deshpande
et al., 2009 [71];
Italy; three years
follow-up
Population-based study
involving 660 older
adults aged 65+ years
Walking while
talking task
MMSE (general cognition)
Only fast gait speed
predicted general
cognitive decline
EXIT 15: 15-item Executive Interview; TMT: Block Design Test, Trail-Making Test; CDR: Clinical Dementia Rating Scale; CLOX 1: Clock-
Drawing Task; CSI-D: Community Screening Instrument of Dementia; COWAT: Controlled Word Association Test; DSST: Digit Symbol
Substitution Test; FCSRT: Free and Cued Selective Reminding Test; LFT: Letter Fluency Test; Modified Mini-Mental Status Examination
(3MS); Wechsler Adult Intelligence Scale—Third Edition (WAIS-III). Table 3. Cont. Author, Year,
Country/Region,
STUDY Type
Participant
Characteristics
Physical
Performance
Measurements
Cognitive Function
Measurements
Results
8. Atkinson
et al., 2007 [67];
3+ years
follow-up
2349 men and women
(mean age 75.6 years)
20-m usual
walking speed
3MS (general cognition); ECF;
CLOX; 1EXIT (Executive function)
Lower global cognitive
function and executive
function were associated
with greater gait
speed decline
9. Deshpande
et al., 2009 [71];
Italy; three years
follow-up
Population-based study
involving 660 older
adults aged 65+ years
Walking while
talking task
MMSE (general cognition)
Only fast gait speed
predicted general
cognitive decline
EXIT 15: 15-item Executive Interview; TMT: Block Design Test, Trail-Making Test; CDR: Clinical Dementia Rating Scale; CLOX 1: Clock-
Drawing Task; CSI-D: Community Screening Instrument of Dementia; COWAT: Controlled Word Association Test; DSST: Digit Symbol
Substitution Test; FCSRT: Free and Cued Selective Reminding Test; LFT: Letter Fluency Test; Modified Mini-Mental Status Examination
(3MS); Wechsler Adult Intelligence Scale—Third Edition (WAIS-III). Table 3. Cont. EXIT 15: 15-item Executive Interview; TMT: Block Design Test, Trail-Making Test; CDR: Clinical Dementia Rating Scale; CLOX 1: Clock-
Drawing Task; CSI-D: Community Screening Instrument of Dementia; COWAT: Controlled Word Association Test; DSST: Digit Symbol
Substitution Test; FCSRT: Free and Cued Selective Reminding Test; LFT: Letter Fluency Test; Modified Mini-Mental Status Examination
(3MS); Wechsler Adult Intelligence Scale—Third Edition (WAIS-III). EXIT 15: 15-item Executive Interview; TMT: Block Design Test, Trail-Making Test; CDR: Clinical Dementia Rating Scale; CLOX 1: Clock-
Drawing Task; CSI-D: Community Screening Instrument of Dementia; COWAT: Controlled Word Association Test; DSST: Digit Symbol
Substitution Test; FCSRT: Free and Cued Selective Reminding Test; LFT: Letter Fluency Test; Modified Mini-Mental Status Examination
(3MS); Wechsler Adult Intelligence Scale—Third Edition (WAIS-III). Epidemiological evidence for links between physical and neurological health is needed. 10. Summary and Conclusions Sui et al.,
2020 [37];
Australia
292 men aged 60+
years; population based
4-m walk speed
test
CogState Brief Battery
(psychomotor function, visual
identification/attention, visual
learning and working memory
For every 1 m/s increase
in gait speed, scores for
psychomotor function
were 0.12 lower, attention
0.08 lower and overall
cognitive function 0.49
higher (all better
function)
Longitudinal Studies
6. Buracchio
et al., 2010 [65];
USA; 20 years
follow-up
204 healthy older
adults (58% female)
aged 65+
9.14-m waking
test
MMSE; CDR (identifying
dementia)
Gait speed declined by
0.02 m/s/year for up to
12 years prior to the onset
of MCI, as assessed using
standardised neurologic
examinations
7. Inzitari et al.,
2007 [66];
5 years follow-up
2776 men and women
aged 75–85 years
6-m walking
speed test
DSST (attention and psychomotor
speed)
Gait speed predicts
decline in attention and
psychomotor speed in
the elderly Complex
walking tasks;
reference
walking tasks 4. Martin et al.,
2013 [62];
Australia
422 older people aged
60–85 years
GAITRite
walkway
COWAT, Category Fluency,
Victoria Stroop test, WAIS-III
(Executive function/attention);
WAIS-III (Processing speed); Rey
Complex Figure copy task
(Visuospatial ability); Hopkins
Verbal Learning Test—revised,
generating scores for total
immediate recall, delayed
recall and recognition memory
and a delayed
reproduction after 20 min of the
Rey Complex Figure (memory) CogState Brief Battery
(psychomotor function, visual
identification/attention, visual
learning and working memory CogState Brief Battery
(psychomotor function, visual
identification/attention, visual
learning and working memory Longitudinal Studies MMSE; CDR (identifying
dementia) DSST (attention and psychomotor
speed) 17 of 22 Int. J. Mol. Sci. 2021, 22, 255 Table 3. Cont. Author, Year,
Country/Region,
STUDY Type
Participant
Characteristics
Physical
Performance
Measurements
Cognitive Function
Measurements
Results
8. Atkinson
et al., 2007 [67];
3+ years
follow-up
2349 men and women
(mean age 75.6 years)
20-m usual
walking speed
3MS (general cognition); ECF;
CLOX; 1EXIT (Executive function)
Lower global cognitive
function and executive
function were associated
with greater gait
speed decline
9. 10. Summary and Conclusions The significance of this review is to provide a rationale for conducting further studies and
targeted intervention trials, with a focus of improving the physical and cognitive health of
the ageing population. Author Contributions: Concept and design: S.X.S., L.J.W., J.A.P.; Drafting of the manuscript: S.X.S.;
S.X.S., L.J.W., K.L.H.-K., N.K.H. and J.A.P. critically appraised the manuscript and approved the final
version. All authors have read and agreed to the published version of the manuscript. Funding: S.X.S. was supported by a Deakin Postgraduate Scholarship in conjunction with the
Geelong Medical and Hospital Benefits Association; L.J.W. by an NHMRC Career Development
Fellowship (1064272) and an NHMRC Investigator grant (1174060); K.L.H.-K. by an Alfred Deakin
Postdoctoral Research Fellowship; N.K.H. by a Dean’s Research Postdoctoral Fellowship (Deakin
University). N.K.H. has received funding from the Norman Beischer Foundation. J.A.P. has received
speaker fees from Amgen, Eli Lilly and Sanofi-Aventis and funding from the NHMRC, Barwon Health,
Deakin University, the BUPA Foundation, Osteoporosis Australia, Australia and New Zealand Bone
and Mineral Society, the Geelong Community Foundation, Western Alliance, Amgen and the Norman
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Sphingosine-1-phosphate/S1P Receptors Signaling Modulates Cell Migration in Human Bone Marrow-Derived Mesenchymal Stem Cells
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Yaxian Kong
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Hindawi Publishing Corporation
Mediators of Inflammation
Volume 2014, Article ID 565369, 11 pages
http://dx.doi.org/10.1155/2014/565369 Hindawi Publishing Corporation
Mediators of Inflammation
Volume 2014, Article ID 565369, 11 pages
http://dx.doi.org/10.1155/2014/565369 Hindawi Publishing Corporation
Mediators of Inflammation
Volume 2014, Article ID 565369, 11 pages
http://dx.doi.org/10.1155/2014/565369 Hindawi Publishing Corporation
Mediators of Inflammation
Volume 2014, Article ID 565369, 11 pages
http://dx.doi.org/10.1155/2014/565369 Yaxian Kong,1,2 Hong Wang,3 Tao Lin,1,2 and Shuling Wang3
1 Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China
2 Beijing Key Laboratory of Emerging Infectious Diseases, Jingshundongjie 8, Beijing 100015, China
3 Stem Cell Research Center, Peking University, Beijing 100191, China Yaxian Kong,1,2 Hong Wang,3 Tao Lin,1,2 and Shuling Wang3
1 Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China
2 Beijing Key Laboratory of Emerging Infectious Diseases, Jingshundongjie 8, Beijing 100015, China
3 Stem Cell Research Center, Peking University, Beijing 100191, China Correspondence should be addressed to Yaxian Kong; kongyaxian@gmail.com Received 30 April 2014; Accepted 12 July 2014; Published 23 July 2014 Academic Editor: Dezheng Zhao Academic Editor: Dezheng Zhao Copyright © 2014 Yaxian Kong et al. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The recruitment of bone marrow-derived mesenchymal stem cells (BMSCs) to damaged tissues and sites of inflammation is
an essential step for clinical therapy. However, the signals regulating the motility of these cells are still not fully understood. Sphingosine-1-phosphate (S1P), a bioactive sphingolipid metabolite, is known to have a variety of biological effects on various
cells. Here, we investigated the roles of S1P and S1P receptors (S1PRs) in migration of human BMSCs. We found that S1P exerted a
powerful migratory action on human BMSCs. Moreover, by employing RNA interference technology and pharmacological tools,
we demonstrated that S1PR1 and S1PR3 are responsible for S1P-induced migration of human BMSCs. In contrast, S1PR2 mediates
the inhibition of migration. Additionally, we explored the downstream signaling pathway of the S1P/S1PRs axis and found that
activation of S1PR1 or S1PR3 increased migration of human BMSCs through a G𝑖/extracellular regulated protein kinases 1/2-
(ERK1/2-) dependent pathway, whereas activation of S1PR2 decreased migration through the Rho/Rho-associated protein kinase
(ROCK) pathway. In conclusion, we reveal that the S1P/S1PRs signaling axis regulates the migration of human BMSCs via a dual-
directional mechanism. Thus, selective modulation of S1PR’s activity on human BMSCs may provide an effective approach to
immunotherapy or tissue regeneration. 1. Introduction Previous studies
by us and other researchers have demonstrated that S1P
strongly stimulated BM cells migration and induced morpho-
logical rearrangements in mice [19–21]. However, the effects
of S1P signaling on human BMSCs and the mechanisms
that regulate their chemotactic behavior are incompletely
understood.f 2.3. Flow Cytometry Analysis. The surface markers of human
BMSCs were analyzed by flow cytometry. Cells were har-
vested by trypsinization, washed once with PBS, and resus-
pended in PBS containing 2% FBS. The cells were incu-
bated with the conjugated antibodies for 30 min on ice. The following antibodies against human antigens were used:
phycoerythrin- (PE-) conjugated anti-CD34, anti-CD45,
anti-CD73, anti-CD105, and anti-CD166 and fluorescein
isothiocyanate- (FITC-) conjugated anti-CD14 and anti-
CD44. Corresponding isotype-matched control monoclonal
antibodies were used in all flow cytometric staining proce-
dures. Flow cytometric analysis was performed using a FACS
Calibur (BD Biosciences, San Diego, CA, USA). In the present study, we characterized the different
effects of S1P receptors on S1P-mediated migration of human
BMSCs and investigated the downstream signaling pathway
in this process. Understanding the role of S1P/S1PRs on
human BMSC migration will promote their effective use for
immunotherapy or tissue regeneration. 2.4. Western Blot Analysis. To monitor the activation of
ERK1/2, cells were exposed to S1P (1 𝜇M) for various times
(2–120 min). Where indicated, cells were pretreated with
S1PR antagonists for 1 h before S1P stimulation (1 𝜇M, 2 min). Cells were lysed in lysis buffer (50 mM Tris-Cl pH 8.0,
150 mM NaCl, 1% Nonidet P-40, 0.1% sodium dodecyl sul-
fate (SDS), 1% Triton X-100, 1 mM sodium orthovanadate,
1 mM phenylmethylsulfonyl fluoride (PMSF), and 10 𝜇g/mL
aprotinin/leupeptin) containing protease inhibitors. Total
protein concentration was determined by Bradford assay. Equal amounts (30 mg) of total protein were separated by 10%
SDS-polyacrylamide gels (SDS-PAGE). For analysis of S1PR1-
3 expression, cells were lysed in lysis buffer as described
before. Protein samples (100 mg) were separated by 10% SDS-
PAGE. Proteins were later transferred from polyacrylamide
gel to methanol-soaked Immobilon polyvinylidene difluoride
membranes (Millipore, Bedford, MA). Primary antibody
incubation was performed overnight at 4∘C. The membranes
were then washed three times and incubated with appropriate
HRP-conjugated secondary antibodies (1 : 1000, Santa Cruz)
at room temperature for 1 h. The membranes were then
washed three times and the signals were visualized using an
enhanced chemiluminescence (ECL Plus) assay kit (Perkin
Elmer, Boston, MA, USA). 1. Introduction The bands were quantified using
GeneSnap software (Perkin Elmer). 1. Introduction and clinical studies, the engraftment of BMSCs into damaged
tissues via migration to suppress immune responses or
enhance tissue repair/regeneration is a crucial process for
clinical therapy [13]. For BMSCs to migrate to and target a
specific tissue, they require the right combination of signaling
molecules from the injured tissue and the corresponding
receptors on BMSCs [13, 14].l Mesenchymal stem cells (MSCs) have been shown to affect
both innate and adaptive immune response [1–3]. They have
been identified and isolated from multiple tissues, including
adipose tissue, umbilical cord, bone marrow, muscle, and fetal
liver [4]. Increasing evidence suggests that bone marrow-
derived mesenchymal stem cells (BMSCs) have therapeu-
tic potential due to their immunosuppressive property in
many immunological disorders, including graft-versus-host
disease [5], Crohn’s disease [6], and the prevention of organ
transplantation rejection [7, 8]. Furthermore, many studies
have demonstrated that BMSCs play a critical role in injury
healing. BMSC transplantation is also regarded as a useful
therapeutic strategy in acute tissue injuries of the lung, heart,
liver, and kidney [9–12]. Of note, in all of these preclinical Among the chemokines and inflammatory mediators
known to trigger potent cellular chemotaxis, sphingosine-1-
phosphate (S1P), which is a bioactive sphingolipid metabolite,
is an ideal candidate [15]. S1P is reported to have a variety
of biological effects on cells, including modulation of motil-
ity, proliferation, differentiation, apoptosis, survival, neurite
retraction, angiogenesis, and regulation of immune function
[16, 17]. S1P can act as both an intracellular second messenger
and a ligand for a family of G protein-coupled receptors 2 Mediators of Inflammation 2.3. Flow Cytometry Analysis. The surface markers of human
BMSCs were analyzed by flow cytometry. Cells were har-
vested by trypsinization, washed once with PBS, and resus-
pended in PBS containing 2% FBS. The cells were incu-
bated with the conjugated antibodies for 30 min on ice. The following antibodies against human antigens were used:
phycoerythrin- (PE-) conjugated anti-CD34, anti-CD45,
anti-CD73, anti-CD105, and anti-CD166 and fluorescein
isothiocyanate- (FITC-) conjugated anti-CD14 and anti-
CD44. Corresponding isotype-matched control monoclonal
antibodies were used in all flow cytometric staining proce-
dures. Flow cytometric analysis was performed using a FACS
Calibur (BD Biosciences, San Diego, CA, USA). referred to as S1P receptor types 1–5 (S1P1–5) [18]. The distinct
response of each cell line to S1P varies depending on its
S1PR expression pattern. S1PR1, S1PR2, and S1PR3 specifically
contribute to S1P-induced cell motility [15]. 2. Material and Methods 2.1. Chemicals and Reagents. Minimum essential medium 𝛼
(MEM 𝛼), penicillin, streptomycin, L-glutamine, and trypsin
were purchased from GIBCO (Grand Island, NY, USA). Fetal bovine serum was from Biochrom (Berlin, Germany). Fluorescence-conjugated monoclonal antibodies for CD44,
CD105, CD166, CD73, CD14, CD34, and CD45 were from BD
Pharmingen (San Diego, CA, USA). Rabbit polyclonal anti-
bodies to S1PR1–3, 𝛼-tubulin, phosphorylated-extracellular
regulated protein kinases 1/2 (p-ERK1/2), and total ERK1/2
were from Santa Cruz Biotechnology (San Diego, CA, USA). PCR reagents and probes used for real-time PCR were from
Applied Biosystems (Foster City, CA, USA). S1P and dihydro-
S1P (H2S1P) were from Biomol (Tebu, France). S1PR1 agonist
(SEW2871), S1PR1 antagonist (W146), S1PR2 antagonist (JTE-
013), and S1PR3 antagonist (CAY10444) were from Cayman
Chemical (Ann Arbor, MI, USA). MEK inhibitor U0126 and
Rho kinase inhibitor Y27632 were from Calbiochem (San
Diego, CA, USA). Bovine serum albumin (BSA) and other
common reagents were from Sigma (St. Louis, MO, USA). 2.2. Isolation of Human BMSCs and Peripheral Blood
Mononuclear Cells (PBMCs). Human BMSCs were obtained
from three healthy donors as described previously [22]. Briefly, bone marrow cells were resuspended in modified
Eagle’s medium of alpha containing 10% fetal bovine serum,
100 U/mL penicillin, 100 𝜇g/mL streptomycin, and 2 mM L-
glutamine and plated in a flask at a density of 3 × 105 cells/mL. Nonadherent cells were discarded after cultivation for 48 h. The adherent cells were washed twice and cultured for 10 to
14 days until cell clones formed. Venous blood was obtained
from three healthy donors and was collected directly into
BD-Vacutainer CPT tubes (BD Biosciences). PBMCs were
isolated by centrifugation according to the manufacturer’s
instructions. The viability of PBMCs was determined by
trypan blue staining and was found to be >90%. Permissions
to use human tissue and blood were granted by Ethical
Committee of Peking University Health Science Center and
Beijing Ditan Hospital, Capital Medical University. 2.5. Migration Assay. In vitro migration was evaluated using
a transwell chamber assay (Millipore, Billerica, MA, USA)
as previously described [21]. In brief, BMSCs were serum-
starved overnight. Where indicated, cells were transfected
with S1PR1-3 siRNA 48 h prior to S1P stimulation (1 𝜇M,
5 min) or pretreated for 1 h with W146 (S1PR1 antagonist),
JTE-013 (S1PR2 antagonist) or CAY10444 (S1PR3 antagonist). Then, 4 × 104 cells were seeded to the upper chamber. 2. Material and Methods Transient transfection of siRNA (40 nM) was per-
formed using Lipofectamine RNAiMAX (Invitrogen, Carls-
bad, CA, USA) as recommended by the manufacturer. Con-
trol cells were treated with 40 nM RNAi Negative Control
Duplexes (scramble siRNA). After 48 h, the transfected cells
were used to perform migration assays. 3.3. S1PR1 and S1PR3 Mediate Promotion of Migration in
Human BMSCs. S1P has been reported to either promote
or inhibit cellular migration, depending on the cell type
examined, via different receptors. Therefore, a series of
techniques were employed to explore the unique effects of
S1P receptors on the migration of human BMSCs. First, we
used siRNA technology to knock down S1PR1 and S1PR3
expression in human BMSCs. To validate this approach, the
mRNA levels of S1PR1 and S1PR3 in cells treated with siRNA
were measured at 48 h after transfection. Human BMSCs
transfected with siRNA targeting S1PR1 or S1PR3 showed
a marked reduction in S1PR1 or S1PR3, whereas the two
siRNAs did not alter the expression of other S1PRs, which
confirmed their specificity (Figures 3(a) and 3(c)). Silencing
of S1PR1 or S1PR3 expression by siRNA effectively attenuated
the migratory effect induced by S1P (Figures 3(d) and 3(f)). Moreover, transfection with a combination of S1PR1 and
S1PR3 siRNA completely abrogated S1P-mediated migration
(Figure 3(f)). 2.8. RhoA Activation Assay. RhoA activation was assessed
using the Rho Assay Reagent (Millipore) according to the
manufacturer’s procedure and as described previously [19]. Briefly, human BMSCs were grown to approximately 80%
confluence on 100 mm dishes and serum-starved in MEM 𝛼
for 12 h and then treated with or without the above-described
S1PRs antagonists before they were stimulated with 1 𝜇M S1P. Immediately after stimulation for the indicated time, cells
were rinsed with cold PBS, lysed in 300 𝜇L of lysis buffer
(25 mM HEPES, pH 7.5, 150 mM NaCl, 1% NP-40, 10 mM
MgCl2, 1 mM EDTA, 2% glycerol, 25 mM NaF, 1 mM sodium
orthovanadate, 1 mM phenylmethylsulfonyl fluoride (PMSF),
and 10 𝜇g/mL aprotinin/leupeptin), and then briefly cen-
trifuged to remove cell debris. Cell lysates were pulled down
by incubation at 4∘C with 20 𝜇g of recombinant Rhotekin-
binding domain bound to glutathione-agarose beads for 1 h. Following washing, bound Rho was eluted by SDS sample
buffer. The eluted samples and the total cell lysates were then
subjected to western blot analysis with RhoA antibody to
detect active and total RhoA, respectively. 2. Material and Methods Various concentrations of S1P, H2S1P, or SEW2871 (S1PR1
agonist) were added to the lower chamber. The chambers
were incubated for 4 h at 37∘C in 5% CO2. Migrated cells,
which adhered to the lower face of the porous membrane,
were fixed with methanol at 4∘C for 1 h and stained with
hematein staining solution. Unmigrated cells on the upper
membrane surface were removed with a cotton swab. The
migration was quantified by analyzing at least six random
fields per filter for each independent experiment. 3 Mediators of Inflammation detectable in human BMSCs in mRNA and protein level
(Figures 1(b) and 1(c)). detectable in human BMSCs in mRNA and protein level
(Figures 1(b) and 1(c)). 2.6. Real-Time PCR. Total RNA was extracted from cells
using the RNeasy kit (Qiagen, Hilden, Germany). cDNA
was synthesized from the total RNA sample using High
Capacity cDNA Reverse Transcription Kits (Applied Biosys-
tems). Real-time PCR was performed in an ABI Prism
7500 sequence detection system (Applied Biosystems). The
expression of the gene of interest was calculated relative to
the levels of GAPDH mRNA (Δct). The expression levels are
presented as “fold change” relative to values obtained with the
control (set as “onefold”). 3.2. S1P Induces Human BMSC Migration through Cell Surface
Receptors. To investigate the chemotaxis of human BMSCs
in response to various concentrations of S1P, we used the
transwell assay and found that low concentrations of S1P
(1–10 nM) exerted a strong dose-dependent migration effect
(Figures 2(a)–2(c)). Meanwhile, higher concentrations of S1P
were less effective and even inhibitory (Figures 2(b) and 2(c)). f
Since S1P can act as both an intracellular second messen-
ger and a ligand for a family of G protein-coupled receptors,
it was of interest to test whether S1P triggers the migration
of human BMSCs via the receptors or not. Therefore, we per-
formed the same experiments using the structural analogue
of S1P, H2S1P, which is only able to mediate its effects through
surface-bound S1PRs [23]. As expected, H2S1P completely
mimicked the induced migration activity of S1P on human
BMSCs (Figure 2(b)), which suggested that S1P induced these
actions via activation of membrane S1PRs. 2.7. RNA Interference. The siRNA sequence targeting specif-
ically human S1PR1–3 was synthesized by Ambion (Grand
Island, NY, USA). BMSCs at 40–50% confluency were pre-
pared. 2. Material and Methods g
To verify this notion, selective S1PRs agonists and/or
antagonists were employed. Human BMSCs displayed a
marked migratory response towards SEW2871, a selective
S1PR1 agonist, in a dose-dependent manner (Figure 4(a)). Moreover, S1P-induced migration of human BMSCs was
completely abrogated by the S1PR1 antagonist W146 or the
S1PR3 antagonist CAY10444, in a dose-dependent manner
(Figures 4(b) and 4(d)). These results indicated that S1PR1
and S1PR3 both contributed to the S1P-induced migration of
human BMSCs. 2.9. Statistical Analysis. Data are expressed as means ± SD
and were analyzed by Student’s t-test when appropriate. 𝑃<
0.05 was considered statistically significant. 3. Results 3.1. Human Bone Marrow-Derived Stem Cells Express S1PR1,
S1PR2, and S1PR3. In line with previous studies, human
BMSCs were confirmed to express CD44, CD105, CD166,
CD73, and lack expression of CD14, CD45, and CD34
(Figure 1(a)). Since S1PR1–3 are the S1P cell surface recep-
tor subtypes that are specifically involved in S1P-mediated
biological activities; we investigated whether these recep-
tors are expressed in human BMSCs. Real-time PCR and
western blot analysis indicated that these receptors were 3.4. S1PR2 Mediates Inhibition of Migration in Human BMSCs. We also used siRNA and pharmacological reagent to evaluate
the effect of S1PR2 on cell migration. The reduced expression
of S1PR2 mRNA in cells treated with S1PR2 siRNA validated
the approach (Figure 3(b)). The siRNA against S1PR2 and
the S1PR2 antagonist JTE-013 both potently enhanced the
S1P-induced migration of BMSCs (Figures 3(e) and 4(c)). In 4 Mediators of Inflammation Mediators of Inflamma
CD44
CD105
CD166
CD14
CD34
CD45
CD73
Counts
Counts
Counts
CD44 PE
CD105 PE
CD166 PE
CD73 PE
CD14 FITC
CD34 FITC
CD45 FITC
0
150
100
101
102
103
104
0
100
101
102
103
104
150
0
100
101
102
103
104
150
Counts
0
150
100
101
102
103
104
Counts
0
150
100
101
102
103
104
Counts
0
150
100
101
102
103
104
Counts
0
150
100
101
102
103
104
(a)
BMSC
PBMC
0.1
1
10
100
Relative mRNA expression
(fold over basal)
S1PR1
S1PR2
S1PR3
(b)
S1PR1
S1PR2
S1PR3
S1P-Rs
𝛼-Tubulin
(c)
gure 1: Expression of S1PRs in BMSCs. (a) The identification of BMSCs was performed by flow cytometry analysis. (b) Real-time
alysis for expression of S1PR1–3 in BMSCs. Human PBMCs as a positive control. (c) Western blot analysis for expression of S1PR1
MSCs. reement with many previous reports, these data demon-
ate that S1PR2 negatively regulated migration mediated by
3.5. S1P-Induced Migration Is Gi Dependent, and the ER
Pathway Is Involved in This Process. 3. Results Since receptors for CD44
CD105
CD166
CD14
CD34
CD45
CD73
Counts
Counts
Counts
CD44 PE
CD105 PE
CD166 PE
CD73 PE
CD14 FITC
CD34 FITC
CD45 FITC
0
150
100
101
102
103
104
0
100
101
102
103
104
150
0
100
101
102
103
104
150
Counts
0
150
100
101
102
103
104
Counts
0
150
100
101
102
103
104
Counts
0
150
100
101
102
103
104
Counts
0
150
100
101
102
103
104
( ) Counts Counts CD14 FITC CD45
CD45 FITC
Counts
0
150
100
101
102
103
104 (a)
BMSC
PBMC
0.1
1
10
100
Relative mRNA expression
(fold over basal)
S1PR1
S1PR2
S1PR3
(b)
S1PR1
S1PR2
S1PR3
S1P-Rs
𝛼-Tubulin
(c)
Figure 1: Expression of S1PRs in BMSCs. (a) The identification of BMSCs was performed by flow cytometry analysis. (b) Real-time P
analysis for expression of S1PR1–3 in BMSCs. Human PBMCs as a positive control. (c) Western blot analysis for expression of S1PR1–
BMSCs. (a) (c) (b) Figure 1: Expression of S1PRs in BMSCs. (a) The identification of BMSCs was performed by flow cytometry analysis. (b) Real-time PCR
analysis for expression of S1PR1–3 in BMSCs. Human PBMCs as a positive control. (c) Western blot analysis for expression of S1PR1–3 in
BMSCs. 3.5. S1P-Induced Migration Is Gi Dependent, and the ERK1/2
Pathway Is Involved in This Process. Since receptors for S1P
are coupled to PTX-sensitive G𝑖as well as PTX-insensitive G
proteins such as G𝑞and/or G12/13 proteins [24], we performed
experiments with cells pretreated with PTX. PTX completely
blocked migration mediated by S1P (Figure 5(a)), which agreement with many previous reports, these data demon-
strate that S1PR2 negatively regulated migration mediated by
S1P in human BMSCs. Taken together, these results demonstrate that S1PR1
and S1PR3, but not S1PR2, are responsible for S1P-induced
migration of human BMSCs. 5 5 Mediators of Inflammation BSA
S1P
(a)
0
0.1
1
10
50
100 1000
0
20
40
60
80
100
Migrated cells/field
S1P
H2S1P
(b)
0
0.1
1
10
50
100
1000
0
5
10
15
20
(nM)
Migration index
(fold over basal)
S1P
H2S1P
(c)
Figure 2: S1P-induced migration of human BMSCs via cell surface receptor. (a) The representative images of serum-starved BMSC migration
stimulated with BSA or 1 nM S1P for 4 h. 3. Results (b)-(c) Serum-starved BMSCs were allowed to migrate for 4 h in the presence of varying
concentrations of S1P and H2S1P, as indicated. Migrated cells in a random fields (b) or migration index (fold over basal, (c)) shown were
counted in 10 random fields per filter for each condition. Data are presented as the mean ± SD. ∗𝑃< 0.05, compared with control. BSA
S1P
(a) S1P S1P 0
0.1
1
10
50
100 1000
0
20
40
60
80
100
Migrated cells/field
S1P
H2S1P
(b) 0
0.1
1
10
50
100
1000
0
5
10
15
20
(nM)
Migration index
(fold over basal) (b) (c) Figure 2: S1P-induced migration of human BMSCs via cell surface receptor. (a) The representative images of serum-starved BMSC migration
stimulated with BSA or 1 nM S1P for 4 h. (b)-(c) Serum-starved BMSCs were allowed to migrate for 4 h in the presence of varying
concentrations of S1P and H2S1P, as indicated. Migrated cells in a random fields (b) or migration index (fold over basal, (c)) shown were
counted in 10 random fields per filter for each condition. Data are presented as the mean ± SD. ∗𝑃< 0.05, compared with control. S1PR2 antagonist JTE-013 did not exert a similar effect
(Figure 5(e)). suggested that S1P-induced migration in human BMSCs was
G𝑖protein dependent. Taken together, these data indicate that S1P induces
human BMSC migration through a G𝑖/ERK-dependent path-
way via S1PR1 and S1PR3. Signaling through ERK1/2 activation has been shown to
mediate S1P-induced migration [19, 25, 26]. Indeed, we found
that S1P induced rapid and transient phosphorylation of
ERK1/2 in a time-dependent manner (Figure 5(b)), indicat-
ing the activation of the ERK1/2 pathway. Treatment of these
cells with U0126, a specific inhibitor of ERK1/2 phosphoryla-
tion, strongly blocked S1P-mediated migration (Figure 5(c)). Previous studies reported the involvement of PTX-sensitive
G𝑖proteins in S1P receptors signal transduction [23, 24] and
that 𝛽𝛾subunits from G𝑖proteins can induce activation of
ERK1/2 [27]. This prompted us to investigate the effect of
PTX on S1P-induced ERK1/2 activation in human BMSCs. As shown in Figure 5(d), pretreatment with PTX blocked the
activation of ERK. 3.6. Rho/Rho-Associated Protein Kinase (ROCK) Pathway
Participates in S1PR2-Mediated Inhibition of Human BMSC
Migration. Pull-down assay showed that RhoA was rapidly
and consistently activated by S1P (Figure 6(a)). It has been
shown that activated GTP-bound Rho activates several
downstream signaling pathways, among which ROCK is a
prominent player [28]. 3. Results Therefore, we tested the effects of a
ROCK inhibitor, Y27632, on BMSC migration and found
that cells treated with Y27632 were considerably augmented
in S1P-induced migration of BMSCs (Figure 6(b)). Taking
our previous data involving S1PR2 into consideration, we
further investigated whether S1PR2, which couples to G12/13,
correlated with the activation of RhoA. As Figure 6(a) shows,
S1PR2 antagonist JTE-013 treatment of cells completely
blocked S1P-mediated activation of Rho, whereas S1PR1
antagonist W146 and S1PR3 antagonist CAY10444 did not 3.6. Rho/Rho-Associated Protein Kinase (ROCK) Pathway
Participates in S1PR2-Mediated Inhibition of Human BMSC
Migration. Pull-down assay showed that RhoA was rapidly
and consistently activated by S1P (Figure 6(a)). It has been
shown that activated GTP-bound Rho activates several
downstream signaling pathways, among which ROCK is a
prominent player [28]. Therefore, we tested the effects of a
ROCK inhibitor, Y27632, on BMSC migration and found
that cells treated with Y27632 were considerably augmented
in S1P-induced migration of BMSCs (Figure 6(b)). Taking
our previous data involving S1PR2 into consideration, we
further investigated whether S1PR2, which couples to G12/13,
correlated with the activation of RhoA. As Figure 6(a) shows,
S1PR2 antagonist JTE-013 treatment of cells completely
blocked S1P-mediated activation of Rho, whereas S1PR1
antagonist W146 and S1PR3 antagonist CAY10444 did not Furthermore, the S1PR1 is known to couple to G𝑖, whereas
S1PR2 and S1PR3 couple to G12/13, G𝑞, and G𝑖[24]. Thus, to
investigate whether blocking of S1PR1, S1PR2, or S1PR3 could
antagonize the phosphorylation of ERK1/2, S1P receptors
antagonists were applied to human BMSCs. 3. Results S1PR1 antagonist
W146 and S1PR3 antagonist CAY10444 treatment completely
abrogated S1P-mediated ERK1/2 phosphorylation, whereas 6 Mediators of Inflammation 6 0
0.2
0.4
0.6
0.8
1.0
1.2
S1PR1
Relative mRNA expression
(fold over basal)
S1PR2
S1PR3
NS siRNA
S1PR1 siRNA
∗
(a)
0
0.2
0.4
0.6
0.8
1.0
1.2
Relative mRNA expression
(fold over basal)
S1PR1
S1PR2
S1PR3
NS siRNA
S1PR2 siRNA
∗
(b)
Relative mRNA expression
(fold over basal)
0
0.2
0.4
0.6
0.8
1.0
1.2
S1PR1
S1PR2
S1PR3
NS siRNA
S1PR3 siRNA
∗
(c)
0
1
2
3
4
5
6
BSA
S1P
NS siRNA
S1PR1 siRNA
Migration index
(fold over control)
∗
(d)
0
2
4
6
8
10
12
Migration index
(fold over control)
NS siRNA
S1PR2 siRNA
BSA
S1P
∗
(e)
0
1
2
3
4
5
6
Control siRNA
S1PR3 siRNA
Migration index
(fold over control)
BSA
S1P
S1PR1+ S1PR3 siRNA
∗
∗
(f)
Figure 3: The effect of silencing S1PR expression on S1P-induced migration in human BMSCs. (a)–(c) Cells were transfected with control
siRNA or with siRNA targeted against S1PR1 (a), S1PR2 (b), or S1PR3 (c) for 48 h. Then S1PR1, S1PR2, or S1PR3 mRNA was evaluated by
real-time RT-PCR. (d)–(f) Effect of silencing S1PR1, S1PR2, or S1PR3 expression on human BMSCs migration in response to S1P. Data are
presented as the mean ± SD. ∗𝑃< 0.05, compared with control siRNA. 0
0.2
0.4
0.6
0.8
1.0
1.2
S1PR1
Relative mRNA expression
(fold over basal)
S1PR2
S1PR3
∗
0
0.2
0.4
0.6
0.8
1.0
1.2
Relative mRNA expression
(fold over basal)
S1PR1
S1PR2
S1PR3
∗
Relative mRNA expression
(fold over basal)
0
0.2
0.4
0.6
0.8
1.0
1.2
S1PR1
S1PR2
S1PR3
∗ 0
1
2
3
4
5
6
BSA
S1P
NS siRNA
S1PR1 siRNA
Migration index
(fold over control)
∗
(d)
0
2
4
6
8
10
12
Migration index
(fold over control)
NS siRNA
S1PR2 siRNA
BSA
S1P
∗
(e)
0
1
2
3
4
5
6
Migration index
(fold over control)
Figure 3: The effect of silencing S1PR expression on S1P-induced migration in human BMSCs. (a
siRNA or with siRNA targeted against S1PR1 (a) S1PR2 (b) or S1PR3 (c) for 48 h Then S1PR1 S Control siRNA
S1PR3 siRNA
S1PR1+ S1PR3 siRNA
(f) NS siRNA
S1PR2 siRNA
(e) (d) (f) (e) Figure 3: The effect of silencing S1PR expression on S1P-induced migration in human BMSCs. 3. Results (a)–(c) Cells were transfected with control
siRNA or with siRNA targeted against S1PR1 (a), S1PR2 (b), or S1PR3 (c) for 48 h. Then S1PR1, S1PR2, or S1PR3 mRNA was evaluated by
real-time RT-PCR. (d)–(f) Effect of silencing S1PR1, S1PR2, or S1PR3 expression on human BMSCs migration in response to S1P. Data are
presented as the mean ± SD. ∗𝑃< 0.05, compared with control siRNA. have such an effect. These data suggest that S1P activates Rho
via S1PR2 and that downstream ROCK activity is required for
the inhibition of migration. effect regulated by the S1P/S1PRs axis which has implications
for the initiation of immunotherapy or tissue regeneration. Our results suggest that S1PR1 and S1PR3 play an important
role in S1P-induced migration of human BMSCs, whereas
S1PR2 negatively regulates S1P-induced migration in human
BMSCs. Furthermore, G𝑖-dependent activation of ERK1/2 as
well as the Rho/ROCK pathway is involved in this process. 4. Discussion 0
1
2
3
4
5
10
50
100
Control
Migration index
(fold over basal)
∗
∗
∗
Sew2871 (𝜇M)
0
1
2
3
4
5
BSA
S1PR1
Migration index
(fold over control)
∗
∗
No W146
1𝜇M W146
10𝜇M W146 (b)
0
1
2
3
4
5
BSA
S1PR3
Migration index
(fold over control)
∗
∗ (a) No CAY10444
1𝜇M CAY10444
5𝜇M CAY10444
(d) (d) (c) Figure 4: The effect of S1PRs agonist or antagonist on S1P-induced human BMSCs migration. (a) Serum-starved cells were allowed to migrate
for 4 h in the presence of the indicated concentration of SEW2871, an S1PR1 agonist. (b)–(d) Serum-starved cells were pretreated for 1 h with
or without the S1PR1 antagonist W146 (b), S1PR2 antagonist JTE-013 (c), and S1PR3 antagonist CAY10444 (d). Pretreated cells were then
allowed to migrate in the presence of S1P. Data are presented as the mean ± SD. ∗𝑃< 0.05, compared with control. to G12/13 leads to inhibition of cell migration [24]. In this
study, S1P-induced migration of BMSCs via S1PR1 and S1PR3
was observed to be sensitive to PTX, thereby implicating
G𝑖-linked signaling pathways. Indeed, the S1PR1 is known
to couple to G𝑖, whereas S1PR2 and S1PR3 couple to G𝑖,
G𝑞, and G12/13 [24, 39]. S1PR1 is a G𝑖-coupled receptor that
stimulates migration in mouse embryonic fibroblasts [33]
and lymphocytes [34, 40]. In contrast, S1PR2, coupled more
strongly to G12/13, is known to inhibit migration in other
cell types, like vascular smooth muscle cells [41] and glioma
cells [38]. However, which G proteins are involved in S1PR3-
mediated cell motility remained unknown. As discussed,
S1PR3 exhibited stimulatory effects in human myofibroblast
[36] and LX-2 cells [35] but had no effects in lymphocytes
[34, 40]. Since S1PR3 couples to members of the G𝑖, G𝑞, and
G12/13 families, such differences are probably attributed to
the varying affinities for the subsets of G proteins found in
different cell types. In the present work, we observed that chemotaxis through S1PR2 [37]. There are some possible
explanations for these contradictory results. First, the gelatin
migration assay which was used in the human lung fibrob-
last studies shows an invasive effect, but not chemotaxis
activity. Thus, it is unsurprising that the invasion assay
displayed opposite results to the transwell assay by our group
and others. 4. Discussion S1P is a bioactive lipid released by many cells during inflam-
mation and following injury which can regulate different
cellular functions [16]. Moreover, interaction between S1P
and its five specific G protein-coupled receptors is known
to regulate many physiological and pathophysiological pro-
cesses including cancer, inflammation, diabetes, and several
immune disorders [18]. One of the profound activities of
extracellular S1P is to mediate cell migration, which is the
focus of this study. S1P-triggered migration of MSCs to sites
of injured tissues is required for cell therapy and tissue
reconstitution. In the present study, we described a potent Along with previous studies in other cell types, S1P was
demonstrated to serve as a good candidate for the induction
of human BMSC motility, and S1PR1–3 were shown to be
specifically involved in this action [29–34]. By employing
pharmacologic tools and RNA interference technology, we
further identified S1PR1/3-mediated stimulatory and S1PR2-
mediated inhibitory signaling in S1P-induced migration of
human BMSCs, which was similar to the phenomenon in
LX-2 cells [35] and human myofibroblasts [36]. However,
conflicting results were found in studies on human lung
fibroblasts which indicated that S1P potentiates fibroblast 7 Mediators of Inflammation 7 Mediators of Inflammation
7
0
1
2
3
4
5
10
50
100
Control
Migration index
(fold over basal)
∗
∗
∗
Sew2871 (𝜇M)
(a)
0
1
2
3
4
5
BSA
S1PR1
Migration index
(fold over control)
∗
∗
No W146
1𝜇M W146
10𝜇M W146
(b)
Migration index
(fold over control)
BSA
S1P
0
2
4
6
8
10
12
∗
No JTE013
1𝜇M JTE013
10𝜇M JTE013
(c)
0
1
2
3
4
5
BSA
S1PR3
Migration index
(fold over control)
∗
∗
No CAY10444
1𝜇M CAY10444
5𝜇M CAY10444
(d)
igure 4: The effect of S1PRs agonist or antagonist on S1P-induced human BMSCs migration. (a) Serum-starved cells were allowed to migrate
or 4 h in the presence of the indicated concentration of SEW2871, an S1PR1 agonist. (b)–(d) Serum-starved cells were pretreated for 1 h with
r without the S1PR1 antagonist W146 (b), S1PR2 antagonist JTE-013 (c), and S1PR3 antagonist CAY10444 (d). Pretreated cells were then
llowed to migrate in the presence of S1P. Data are presented as the mean ± SD. ∗𝑃< 0.05, compared with control. 4. Discussion In addition, S1PR2 was reported to decrease
glioma cell motility but enhance invasion through inducing
cell interaction with the extracellular matrix and matrix
degradation in glioma cell [38]. Therefore, S1PR2-mediated
interaction with matrix might play an important role in these
contradictory studies. With regards to the role of S1PR2
in invasion versus chemotaxis, more detailed mechanisms
should be studied.hf The varied cellular effects of S1P were mainly attributed
to the coupling of these receptors to different G proteins
and their differential signaling cascades. In particular, S1P
receptor coupling to G𝑖leads to cell migration, while coupling 8
Mediators of Inflammation
0
1
2
3
4
5
6
7
No PTX
Migration index
(fold over control)
BSA
S1P
2.5 ng/mL PTX
5.0ng/mL PTX
∗
∗
(a)
P-Erk
Total-Erk
(min)
120
60
30
10
5
2
0
(b)
BSA
S1P
Migration index
(fold over control)
No U0126
0
1
2
3
4
5
1𝜇M U0126
10𝜇M U0126
∗
∗
(c)
PTX
S1P
P-Erk
Total-Erk
+
+
+
−
(d)
ctrl
DMSO
ctrl
DMSO W146
JTE
CAY
S1P
P-Erk
Total-Erk
(e)
Figure 5: Involvement of the PTX-sensitive ERK1/2 pathway in S1P-induced migration of human BMSCs. (a), (c) Serum-starved BMSCs
were pretreated with the indicated concentration of PTX (a) or U0126 (c) and then were allowed to migrate for 4 h in the presence of 1 nM
S1P. (b) The effect of S1P on the activation of ERK1/2 MAPK. Cells were stimulated with S1P (1 𝜇M) for the indicated time and cell lysates were
analyzed by western blot. (d)-(e): The effect of S1PRs antagonists (S1PR1 antagonist W146, S1PR2 antagonist JTE-013, and S1PR3 antagonist
CAY1444) and PTX on S1P-induced activation of ERK1/2. Cells were pretreated with S1PRs antagonists (d) or pretreated with 5 ng/mL PTX
(e). The pretreated cells were stimulated with 1 𝜇M S1P for 2 min, and cell lysates were analyzed by western blot. Data are presented as the
mean ± SD. ∗𝑃< 0.05, compared with control. Mediators of Inflammation 8 No U0126
1𝜇M U0126
10𝜇M U0126
(c) (e) (e) (d) (c) Figure 5: Involvement of the PTX-sensitive ERK1/2 pathway in S1P-induced migration of human BMSCs. (a), (c) Serum-starved BMSCs
were pretreated with the indicated concentration of PTX (a) or U0126 (c) and then were allowed to migrate for 4 h in the presence of 1 nM
S1P. (b) The effect of S1P on the activation of ERK1/2 MAPK. 4. Discussion Cells were stimulated with S1P (1 𝜇M) for the indicated time and cell lysates were
analyzed by western blot. (d)-(e): The effect of S1PRs antagonists (S1PR1 antagonist W146, S1PR2 antagonist JTE-013, and S1PR3 antagonist
CAY1444) and PTX on S1P-induced activation of ERK1/2. Cells were pretreated with S1PRs antagonists (d) or pretreated with 5 ng/mL PTX
(e). The pretreated cells were stimulated with 1 𝜇M S1P for 2 min, and cell lysates were analyzed by western blot. Data are presented as the
mean ± SD. ∗𝑃< 0.05, compared with control. contributed to chemotaxis towards S1P in human BMSCs. Our finding that ERK activation is a result of S1P signaling
through S1PR1 and S1PR3 is in agreement with previous
studies in other cells. For example, NADPH oxidase activity
and intracellular H2O2 levels increase in NIH3T3 fibroblasts
as a result of activated ERK caused by S1P signaling [46]. However, there are also reports of S1PR3 exerting no effects
on ERK1/2 activation, but instead inducing the activation of
Akt [44]. Furthermore, S1PR2, a receptor often exhibiting
inhibitory effects on migration, can lead to the activation of
ERK1/2 in primary rat hepatocytes [45]. The reason for such
discrepancies is not clear yet, although different cell types
used in these studies may be one of the causes. S1PR3 stimulated migration as well as S1PR1, which implies
that S1PR3 coupled more effectively to G𝑖than G𝑞, and
G12/13 in human BMSCs. Thus, a further insight into the
mechanistic details of S1P/S1PRs/G protein signaling axis is
necessary to understand human BMSCs migration.h y
g
The MAPK/ERK pathway has been widely demonstrated
to play an essential role in cell survival, proliferation, and
migration [25, 26, 42]. Indeed, S1P-mediated cell motility
was regulated by the MAPK signal transduction pathway in
different cell types. In particular, ERK1/2 activation was often
associated with an S1PR1-G𝑖-dependent pathway [26, 43, 44]. Additionally, S1PR2 and S1PR3 were also observed to induce
rapid and reversible S1P-mediated ERK phosphorylation [45–
48]. However, which S1P receptor subtype leads to activation
of ERK and cell migration in human BMSCs remained
unknown. Here, we found that both S1PR1 and S1PR3,
but not S1PR2, were required for activation of ERK and In addition, S1PR2 coupling with G12/13 was demon-
strated to result in activation of Rho with subsequent inhi-
bition of cell motility. Conflict of Interests The authors declare that there is no conflict of interests
regarding the publication of this paper. Figure 6: The essential role of the Rho pathway in S1PR2-mediated
suppression of human BMSCs migration. (a) Left: cells were stim-
ulated with S1P (1 𝜇M) for the indicated time. Right: cells were
pretreated with S1PRs antagonists (S1PR1 antagonist W146, S1PR2
antagonist JTE-013, and S1PR3 antagonist CAY1444) and then were
stimulated with 1 𝜇M S1P for 5 min. RhoA activities were determined
by a pull-down assay. (b) Serum-starved BMSCs were pretreated
with the indicated concentration of Y27632 and then were allowed
to migrate for 4 h in the presence of 1 nM S1P. Acknowledgments Therefore, migration mediated
by S1P is a complicated process which requires a different
activation status of the actin cytoskeleton, including gradient Authors’ Contribution Yaxian Kong and Hong Wang contributed equally to this
work. 4. Discussion In accordance with previous findings,
our study suggested that Rho/ROCK activity was required 9 Mediators of Inflammation S1P sensing, polarization, and orientation versus subsequent
migration at different stages of activation. Further studies are
expected to elucidate more mechanistic details involved in the
varied migration of human BMSCs. (min)
120
60
30
10
5
2
0
GTP-RhoA
Total-RhoA
S1P
W146
CAY
JTE
(a)
Migration index
(fold over control)
0
2
4
6
8
10
BSA
S1P
No Y27632
0.1 𝜇M Y27632
1𝜇M Y27632
∗
∗
(b)
Figure 6: The essential role of the Rho pathway in S1PR2-mediated
suppression of human BMSCs migration. (a) Left: cells were stim-
ulated with S1P (1 𝜇M) for the indicated time. Right: cells were
pretreated with S1PRs antagonists (S1PR1 antagonist W146, S1PR2
antagonist JTE-013, and S1PR3 antagonist CAY1444) and then were
stimulated with 1 𝜇M S1P for 5 min. RhoA activities were determined
by a pull-down assay. (b) Serum-starved BMSCs were pretreated
with the indicated concentration of Y27632 and then were allowed
to migrate for 4 h in the presence of 1 nM S1P. (min)
120
60
30
10
5
2
0
GTP-RhoA
Total-RhoA
W146
CAY
JTE
(a) g
In summary, our present study demonstrates that activa-
tion of S1PR1 or S1PR3 increases migration of human BMSCs
by activating of MAPK/ERK pathway via G𝑖protein. In con-
trast, activation of S1PR2 decreases migration via Rho activa-
tion. These results suggest that the coupling of S1P receptors
to various heterotrimeric G proteins and, consequently,
distinct downstream signaling pathways lead to downstream
pathological phenomena. It is noted that S1PR1/3 and S1PR2
as well as the MAPK/ERK and Rho/ROCK pathways help
to balance migration of human BMSCs mediated by S1P. Importantly, our results unravel an important part of the
steps that lead to the activation of directional migration in
BMSCs, and they may allow us to better develop BMSCs as
an improved cellular therapy for immunological disorders or
tissue regeneration. (a) Acknowledgments The authors thank Professor Lingsong Li for his scientific
advice. This work was supported through the Grants 863
Program (2009DFB30300) from the Ministry of Science and
Technology of China. for inhibiting S1P-induced migration in human BMSCs. In
contrast, gelatin migration assay used in some previous
studies exhibited the activation of Rho induced migration
in endothelial cells and bone marrow cells [19, 49]. Meriane
et al. have implied that matrix metalloproteinase-mediated
migration stimulated by S1P in the gelatin migration assay
was associated with an increase in Rho activity and actin
stress fibers [19], whereas the transwell assay displayed
enhanced chemotactic migration corresponding to reduced
Rho activity and stress fibers in bone marrow stromal cells
[50]. As S1P-triggered Rho activation is mainly mediated
by S1PR2, the role for Rho in migration is probably corre-
sponding to S1PR2 [24]. Similar to that S1PR2 exerts opposite
migration effects, Rho-mediated stimulatory invasive activ-
ity, and inhibitory chemotactic effect. In particular, mediation
of stress fibers may confound the interpretation of these
experiments. Interestingly, during chemotaxis, an increase
in stress fiber formation might not be required when S1P
is acting as a chemoattractant. In contrast, actin stress fiber
formation is necessary in invasive migration which involves
matrix degradation [19, 50]. Therefore, migration mediated
by S1P is a complicated process which requires a different
activation status of the actin cytoskeleton, including gradient for inhibiting S1P-induced migration in human BMSCs. In
contrast, gelatin migration assay used in some previous
studies exhibited the activation of Rho induced migration
in endothelial cells and bone marrow cells [19, 49]. Meriane
et al. have implied that matrix metalloproteinase-mediated
migration stimulated by S1P in the gelatin migration assay
was associated with an increase in Rho activity and actin
stress fibers [19], whereas the transwell assay displayed
enhanced chemotactic migration corresponding to reduced
Rho activity and stress fibers in bone marrow stromal cells
[50]. As S1P-triggered Rho activation is mainly mediated
by S1PR2, the role for Rho in migration is probably corre-
sponding to S1PR2 [24]. Similar to that S1PR2 exerts opposite
migration effects, Rho-mediated stimulatory invasive activ-
ity, and inhibitory chemotactic effect. In particular, mediation
of stress fibers may confound the interpretation of these
experiments. Interestingly, during chemotaxis, an increase
in stress fiber formation might not be required when S1P
is acting as a chemoattractant. In contrast, actin stress fiber
formation is necessary in invasive migration which involves
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General rights
Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners
and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. Bond University
Research Repository
Reliability of kettlebell swing one and five repetition maximum
Ross, James A; Keogh, Justin W L; Lorenzen, Christian
Published in:
PEERJ
DOI:
10.7717/peerj.14370
Licence:
CC BY
Link to output in Bond University research repository.
Recommended citation(APA):
Ross, J. A., Keogh, J. W. L., & Lorenzen, C. (2022). Reliability of kettlebell swing one and five repetition
maximum. PEERJ, 10, Article e14370. https://doi.org/10.7717/peerj.14370 Bond University
Research Repository Reliability of kettlebell swing one and five repetition maximum Reliability of kettlebell swing one and five
repetition maximum James A. Ross1, Justin W. L. Keogh2,3,4 and Christian Lorenzen1 1 School of Behavioural and Health Sciences, Australian Catholic University, Melbourne, Victoria,
Australia 2 Faculty of Health Sciences and Medicine, Institute of Health & Sport, Bond University, Gold
Coast, Queensland, Australia 3 Manipal Academy of Higher Education Mangalore, Kasturba Medical College, Manipal,
Karnataka, India 4 Sports Performance Research Centre New Zealand, AUT University, Auckland, New Zealand Background: Research into the kettlebell swing has increased in the last decade. There has been a paucity of literature assessing an individual’s ability to perform the
kettlebell swing exercise. The purpose of this study was to determine the test-retest
reliability of the one and five repetition maximum (1RM and 5RM) kettlebell swing. Materials & Methods: Twenty four recreational resistance-trained participants
performed an isometric mid-thigh pull (IMTP) and two familiarization sessions
followed by three test sessions for each RM load approximately one week apart, using
a custom-built plate-loaded kettlebell. On each test occasion, subjects completed a
series of warm-up sets followed by 3–4 progressively heavier kettlebell swings to a
standardized height until 1RM or 5RM was reached. Test-retest reliability was
calculated using the intra-class correlation (ICC) and typical error was represented as
the coefficient of variation (CV%) with 90% confidence limits (90% CL). The smallest
worthwhile change (SWC%) representing the smallest change of practical importance, was calculated as 0.2 × between-subject standard deviation. The relationship of kettlebell swing performance and maximum strength was
determined by Pearson correlation with ±90% CL between the absolute peak force
recorded during IMTP and 1RM or 5RM. Submitted 8 July 2022
Accepted 19 October 2022
Published 21 November 2022
Corresponding author
James A. Ross,
james.ross33@yahoo.com
Academic editor
Gerald Mangine
Additional Information and
Declarations can be found on
page 9
DOI 10.7717/peerj.14370
Copyright
2022 Ross et al. Distributed under Results: Results demonstrated a high test-retest reliability for both the 1RM
(ICC = 0.97, 90% CL [0.95–0.99]; CV = 2.7%, 90% CL [2.2–3.7%]) and 5RM
(ICC = 0.98, 90% CL [0.96–0.99]; CV = 2.4%, 90% CL [1.9–3.3%]), respectively. The CV% was lower than the SWC for both the 1RM (SWC = 2.8%, 90% CL
[1.9–3.5]) and 5RM (SWC = 2.9%, 90% CL [1.9–3.6]) kettlebell swing. The correlation between IMTP absolute peak force and the 1RM (r = 0.69, 90% CL
0.43–0.83) was large and very large for the 5RM (r = 0.75, 90% CL [0.55–0.87]). Subjects Kinesiology, Biomechanics, Sports Medicine
Keywords Resistance training, Strength training, Exercise, 1RM, 5RM, Test-retest Reliability of kettlebell swing one and five repetition maximum Link to output in Bond University research repository. Recommended citation(APA):
Ross, J. A., Keogh, J. W. L., & Lorenzen, C. (2022). Reliability of kettlebell swing one and five repetition
maximum. PEERJ, 10, Article e14370. https://doi.org/10.7717/peerj.14370 Recommended citation(APA):
Ross, J. A., Keogh, J. W. L., & Lorenzen, C. (2022). Reliability of kettlebell swing one and five repetition
maximum. PEERJ, 10, Article e14370. https://doi.org/10.7717/peerj.14370 General rights
Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners
and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. For more information, or if you believe that this document breaches copyright, please contact the Bond University rese
coordinator. Download date: 25 Oct 2024 Download date: 25 Oct 2024 How to cite this article Ross JA, Keogh JWL, Lorenzen C. 2022. Reliability of kettlebell swing one and five repetition maximum.
PeerJ 10:e14370 DOI 10.7717/peerj.14370 INTRODUCTION The use of kettlebells as resistance training implements has gained increasing popularity
(Meigh et al., 2019). The increased popularity of kettlebells is likely due to their versatility
in allowing an extensive range of exercises to be performed in order to provide a stimulus
for improving a range of muscular strength, power and endurance qualities (Lake &
Lauder, 2012a, 2012b; Wade et al., 2017; Wesley & Kivi, 2017). This increase in popularity
coincided with the commercial range of kettlebells increasing from 4–48 kg (Tsatsouline,
2006) to 2–94 kg (Meigh et al., 2019). The most commonly researched exercise is the
kettlebell swing (Meigh et al., 2019), which is commonly performed with heavier kettlebells
and has been the topic of future research suggestions (Lake & Lauder, 2012b). However,
despite its popularity, little data exists on assessment protocols for this movement. In contrast to kettlebell training, valid and reliable protocols for the assessment for a
range of strength and power movements such as the squat, bench press, and power clean
are well established (McGuigan & Winchester, 2008; Sheppard et al., 2008). These protocols
often involve the determination of the maximum load that can be lifted for a specified
number of repetitions which is known as the repetition maximum (RM) (Grgic et al., 2020;
Reynolds, Gordon & Robergs, 2006). In major compound lifts, RM load is commonly
assessed between 1RM and 5RM but can be conducted at any RM (McMaster et al., 2014). The kettlebell swing has been prescribed with 8–12RM (Lyons et al., 2017) and a 20RM
load (Sørensen et al., 2021). Additionally, less common methods have also been used such
as: percentage of isometric strength (Maulit et al., 2017), loads based on body weight (Lake
& Lauder, 2012a; Levine et al., 2020), expert opinion (Farrar, Mayhew & Koch, 2010), peak
power (Kartages et al., 2019) and Rating of Perceived Exertion (Meigh et al., 2022). These
methods can be used to guide training prescription, however, assessing RM also provides a
method of assessing performance change within a chosen exercise following training
interventions (McMaster et al., 2014). However, in order to do this effectively, the
reliability of RM loads in specific exercises should first be established. Reliability refers to the consistency of results between consecutive tests or performances
(Comfort & McMahon, 2015; Cormack et al., 2008; Hopkins, 2000). Reliability of kettlebell swing one and five
repetition maximum Conclusions: These results demonstrate the stability of 1RM and 5RM kettlebell
swing performance after two familiarization sessions. Practitioners can be confident
that changes in kettlebell swing 1RM and 5RM performance of >3.6 kg represent a
practically important difference, which is the upper limit of the 90% CL. Results: Results demonstrated a high test-retest reliability for both the 1RM
(ICC = 0.97, 90% CL [0.95–0.99]; CV = 2.7%, 90% CL [2.2–3.7%]) and 5RM
(ICC = 0.98, 90% CL [0.96–0.99]; CV = 2.4%, 90% CL [1.9–3.3%]), respectively. The CV% was lower than the SWC for both the 1RM (SWC = 2.8%, 90% CL
[1.9–3.5]) and 5RM (SWC = 2.9%, 90% CL [1.9–3.6]) kettlebell swing. Distributed under
Creative Commons CC-BY 4.0 Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 Participants Twenty four male and three female recreationally resistance-trained participants were
recruited from local gyms with at least one year of resistance and kettlebell training
experience. Twenty four males (age: 31 ± 5 years; training age: 9 ± 5 years; height: 183 ±
7 cm; body mass: 91 ± 13 kg) completed the first 5RM session, however, data collection was
then impeded by the COVID-19 pandemic state restrictions, limiting participation. Therefore, a total of 23 males completed the first 1RM session, and 21 and 20 males
completed all three of the 1RM and 5RM sessions, respectively. Females were excluded
from analysis due to low statistical power. All participants were free of any injury that
would impact performance and provided written, informed consent prior to testing. Ethical approval was granted by the Australian Catholic University Human Research
Ethics Committee (2018-265E). Trial conditions A within-subject test-retest design was used to determine the reliability of the kettlebell
swing 1RM and 5RM in recreationally resistance-trained participants. Participants
attended an isometric strength assessment, two familiarization sessions, and then three
separate sessions for each of the 1RM and 5RM assessments, combining for a total of nine
sessions. The order of the participant’s attendance for the 1RM and 5RM was randomly
assigned. The 1RM and 5RM sessions took place 72 h apart and there was 1 week between
repeat trials of the 1RM and 5RM. The participants could train normally after each RM
session to avoid detraining and minimize any training interference. INTRODUCTION This concept is
important in strength training environments as it provides quantification of the “noise” in
a test (Hopkins, 2000). Noise is the result of both biological and technical variation and can
therefore be used to determine whether any change in performance can be considered
practically important based on whether the inherent noise in the test has been exceeded
(Appleby, Newton & Cormack, 2019). Reliability values, in the form of the Typical Error (TE) and coefficient of variation
(CV%) for common resistance training exercises at a range of RM loads have been
well established (Comfort & McMahon, 2015; Cormack et al., 2008; Hopkins, 2000). For example, values for the bench press and squat are reported as <1 CV% (Seo et al.,
2012), which are somewhat lower than for movements requiring higher skill levels, such as
the power clean 4.8 CV% (Sheppard et al., 2008). Further, the squat correlates nearly
perfectly to isometric strength (r = 0.97) (Mcguigan et al., 2010), yet the kettlebell swings
relationship to isometric strength is unknown. Whilst reliability values are well known for Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 2/13 a range of resistance training exercises, no data exists for the kettlebell swing. Therefore,
the primary aim of this research is to establish the reliability of the 1RM and 5RM kettlebell
swing, and the secondary aim is to determine their relationship to isometric strength. MATERIALS AND METHODS Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 Procedures All sessions included a warm-up involving 5 min of stationary cycling at a self-selected
pace. Isometric strength assessment involved an isometric mid-thigh pull (IMTP). During
the IMTP participants stood on portable force plates (PASCO PS-2142; PASCO 147
scientific, Roseville, CA, USA) and secured their grip with straps upon an immovable bar. When the participants were in the correct position the instruction to pull “hard and fast”
was given (McGuigan & Winchester, 2008). The participants performed three trials of 5 s
with 3 min rest (McGuigan & Winchester, 2008). During the five seconds vertical ground
reaction force was recorded at 1,000 hz. The kettlebell familiarization sessions involved
progressively heavier sets of the kettlebell swing starting with a set of 8–12 repetitions and
finishing with either 4RM or 8RM for the 1RM and 5RM familiarization sessions,
respectively. An estimated 1RM was calculated from the heaviest set of four repetitions
during the warm-up for the 1RM familiarization trial using the following formula:
Estimated 1RM ¼
Weight lifted
1:02780:0278repetitions performed (Brzycki, 1993). Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 3/13 Figure 1 Protocols. Familiarization, 1RM and 5RM assessments and warm-up protocols. Full-size
DOI: 10.7717/peerj.14370/fig-1 Figure 1 Protocols. Familiarization, 1RM and 5RM assessments and warm-up protocols. Full-size
DOI: 10.7717/peerj.14370/fig-1 Figure 1 Protocols. Familiarization, 1RM and 5RM assessments and warm-up protocols. Full-size
DOI: 10.7717/peerj.14370/fig-1 The protocol for the reliability trial sessions is outlined in Fig. 1. Following the 5 min
of stationary cycling, the warm-up sets for the 1RM sessions included 50–60% 1RM for
10 repetitions, 70–80% 1RM for five repetitions, 90% 1RM for 1–3 repetitions from
estimated 1RM. The warm-up for the 5RM sessions involved 40–50% 1RM for 10
repetitions, 60–70% 1RM for five repetitions and 80% 1RM for 1–3 repetitions from
estimated 1RM. Participants completed the 1RM and 5RM using the hip hinge swing style with
self-selected ankle dorsiflexion to accommodate the maximum effort. The kettlebell was
swung with the intention of making contact with a marker set to standing acromion process
height to ensure swing height consistency, (see Fig. 2). The swing trials started in the deadlift
finish position, the first and second repetitions were considered as ‘build up’ swings. The build-up swings allowed increased momentum and displacement to impose a similar
eccentric phase as other repetitions. The third swing was considered the first repetition
attempt for the 1RM and 5RM. Procedures A total of 5 min of rest was allowed between the RM attempts. The criterion for successful kettlebell swing attempts was that the plates of the kettlebell
needed to contact the foam marker, illustrated in Fig. 2. Three to four trials of one or five
repetitions were performed, with the load incrementally increased by 2.5–10 kg after each
successful attempt, this was repeated until 1RM or 5RM was reached (Sheppard et al., 2008). The trial was disregarded and retested with the same load if there was a loss of balance. Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 Instrumentation An adjustable foam measuring marker was used as a target for the 1RM and 5RM
assessments utilizing a plate-loaded kettlebell. The foam marker was free to move up and
down the vertical beam and was secured with spring-loaded clamps. It was set to the top of
the acromion process height and adjusted to accommodate the plate radius. The plate- Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 4/13 Figure 2 Kettlebell swing assessment. Illustrates the starting position, followed by two build up swings
and a successful 1RM kettlebell swing. Full-size
DOI: 10.7717/peerj.14370/fig-2 Figure 2 Kettlebell swing assessment. Illustrates the starting position, followed by two build up swings
and a successful 1RM kettlebell swing. Full-size
DOI: 10.7717/peerj.14370/fig-2 loaded kettlebell had a 35 mm handle with a shaft that was 280 mm long and a diameter of
50 mm. The plate-loaded kettlebell was loaded with 1, 2.5, 5 and 10 kg steel plates (MA1,
Dandenong South, Australia), when loaded on the shaft these plates (Fig. 2) added 12, 12,
14 and 15 mm respectively to the length of the loaded plate stack. These plates were not
calibrated so the kettlebell was weighed on a portable force plate (PASCO PS-2142;
PASCO scientific, Roseville, CA, USA), after the successful 1RM and 5RM trials. loaded kettlebell had a 35 mm handle with a shaft that was 280 mm long and a diameter of
50 mm. The plate-loaded kettlebell was loaded with 1, 2.5, 5 and 10 kg steel plates (MA1,
Dandenong South, Australia), when loaded on the shaft these plates (Fig. 2) added 12, 12,
14 and 15 mm respectively to the length of the loaded plate stack. These plates were not
calibrated so the kettlebell was weighed on a portable force plate (PASCO PS-2142;
PASCO scientific, Roseville, CA, USA), after the successful 1RM and 5RM trials. Statistical analyses A priori power analysis to determine the minimum sample size (n) was performed with R
(version 4.0.2 (2020-06-22) package ‘ICC.Sample.Size’ (Zou, 2012) (input parameters:
(p = 0.90, p0 = 0.73, k = 3, alpha = 0.05, tails = 2, power = 0.80)). Twenty participants were
required in order to detect excellent reliability (ICC ≥0.90). The test-retest reliability
statistical analyses included a consecutive pairwise intraclass correlation coefficient (ICC)
(3, 1) and typical error represented as the coefficient of variation (CV%) ±90% confidence
limits (CL) and difference in mean using a customized Excel spreadsheet (Hopkins, 2017). The ICC was classified by the following scale: ‘poor’ ICC = <0.5, ‘moderate’
ICC = 0.5–0.75, ‘good’ ICC = 0.75–0.9, and ‘excellent’ ICC = >0.9 (Koo & Li, 2016). The smallest worthwhile change (SWC) was determined as 0.2 × between-participants SD
± 90% CL representing the smallest practically important change (Appleby, Cormack &
Newton, 2019; Appleby, Newton & Cormack, 2019; Tofari et al., 2015). If the CV% was
smaller than the SWC, the 1RM and 5RM tests were considered sensitive enough to detect
the SWC. The relationship of kettlebell swing performance and maximum strength was
determined by Pearson correlation with ±90% CL (Hopkins, 2017). The highest absolute
peak force value for the IMTP and the first trial of either the 1RM or 5RM were analyzed. The Pearson correlation coefficient was classified by the following scale: ‘trivial’ r < 0.1,
‘small’ r = 0.1–0.3, ‘moderate’ r = 0.3–0.5, ‘larger’ r = 0.5–0.7, ‘very large’ r = 0.7–0.9, ‘nearly
perfect’ r = 0.9–0.1, ‘perfect’ r = 1 (Hopkins, 2014). A priori power analysis to determine the minimum sample size (n) was performed with R
(version 4.0.2 (2020-06-22) package ‘ICC.Sample.Size’ (Zou, 2012) (input parameters:
(p = 0.90, p0 = 0.73, k = 3, alpha = 0.05, tails = 2, power = 0.80)). Twenty participants were
required in order to detect excellent reliability (ICC ≥0.90). The test-retest reliability
statistical analyses included a consecutive pairwise intraclass correlation coefficient (ICC)
(3, 1) and typical error represented as the coefficient of variation (CV%) ±90% confidence
limits (CL) and difference in mean using a customized Excel spreadsheet (Hopkins, 2017). The ICC was classified by the following scale: ‘poor’ ICC = <0.5, ‘moderate’ RESULTS The mean ± SD loads for each trial of the 1RM and 5RM are displayed in Table 1. Table 2
contains the CV%, ICC and SWC for the comparison of trial 1 vs trial 2 and trial 2 vs trial 3. The CV% values and ICC’s of pairwise comparisons were similar in both the 1RM and
5RM conditions. In all cases the CV% was less than the SWC. The mean ± SD of the IMTP Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 Table 1 The 1RM and 5RM kettlebell swing mean and difference in the mean (kg). Intensity
Trial 1 (kg) (±SD)
Trial 2 (kg) (±SD)
Trial 3 (kg) (±SD)
Mean difference trial
2–1 (kg) (±90% CL)
Mean difference trial
3–2 (kg) (±90% CL)
1RM
73.93 ± 10.52
75.53 ± 11.23
76.50 ± 10.43
1.60 [0.72–2.48]
0.97 [−0.07 to 2.01]
5RM
67.49 ± 10.01
70.67 ± 9.72
70.61 ± 9.66
3.19 [2.34–4.04]
−0.07 [−0.83 to 0.70]
Note:
SD, standard deviation; CL = confidence limits. Table 2 Test-retest ICC, CV% and SWC% for 1RM and 5RM kettlebell swing. Intensity
ICC (±90% CL)
CV% (±90% CL)
SWC% (±90% CL)
ICC (±90% CL)
CV% (±90% CL)
SWC% (±90% CL)
Trial 2–1
Trial 3–2
1RM
0.98 [0.96–0.99]
2.4 [1.9–3.3]
2.8 [1.9–3.5]
0.97 [0.94–0.99]
2.7 [2.2–3.7]
2.8 [1.9–3.5]
5RM
0.98 [0.95–0.99]
2.4 [1.9–3.3]
2.9 [1.9–3.6]
0.98 [0.96–0.99]
2.1 [1.7–2.9]
2.8 [1.9–3.5]
Note:
ICC, intraclass correlation coefficient; CV, coefficient of variation; SWC%, smallest worthwhile change, calculated as a 0.2 times the between-subject pure SD and
represented as a percentage. , coefficient of variation; SWC%, smallest worthwhile change, calculated as a 0.2 times the between-subject pure SD and absolute peak force was 3,555 ± 647N and there was a large (r = 0.69, 90% CL [0.43–0.83])
and very large (r = 0.75, 90% CL [0.55–0.87]) correlation between maximal isometric
strength and the 1RM and 5RM swing, respectively. absolute peak force was 3,555 ± 647N and there was a large (r = 0.69, 90% CL [0.43–0.83])
and very large (r = 0.75, 90% CL [0.55–0.87]) correlation between maximal isometric
strength and the 1RM and 5RM swing, respectively. Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 DISCUSSION The primary aim of this research was to quantify the reliability of the 1RM and 5RM
kettlebell swing. The results suggest that both the 1RM and 5RM kettlebell swing possess
acceptable inter-day reliability following two familiarization trials. Furthermore, the CV%
of the 1RM and 5RM kettlebell swing is less than the SWC, demonstrating that any
variation in 1RM or 5RM performance greater than the SWC represents a practically
important change. The assessment of the maximum load that can be lifted for a specified number of
repetitions is commonplace in strength and conditioning practice (Lawton, Cronin &
McGuigan, 2014). Pre and post-intervention RM assessment is commonly used to
determine changes in the performance of a specific exercise. However, the capacity to use
this information to determine whether a practically important change has occurred from
one test to the next is only possible when the variation or “noise” from both biological and
technical sources has been quantified (Currell & Jeukendrup, 2008). This data is available
for many common protocols across a variety of repetition ranges, including the squat,
bench press and power clean (Comfort & McMahon, 2015; Grgic et al., 2020; Seo et al.,
2012). In general, the “noise” appears to increase with increasing complexity of movement,
but this increase is only slight (Grgic et al., 2020; Sheppard et al., 2008). Whilst the kettlebell
swing may appear to be a somewhat more complex movement to assess than other
multi-joint exercises, the results of the current research demonstrates similar error
(ICC = 0.97, 90% CL [0.94–0.99] and CV = 2.7%, 90% CL [2.1–2.7%]) to that typically
observed in both single joint exercises and more complex tasks such as the power clean
(Comfort & McMahon, 2015; Grgic et al., 2020; Lawton, Cronin & McGuigan, 2014;
McCurdy et al., 2008). Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 6/13 In addition to exercise complexity, factors such as training status, age, body–region, and
sex, have been examined for their impact on reliability and in general, it appears that these
factors have minimal impact (Grgic et al., 2020). The findings of this research are in
agreement with previous work in terms of the impact of familiarization, as reliability
remained largely unchanged when the difference in trial one vs trial two and trial two vs
trial three performance is considered (Banyard, Nosaka & Haff, 2017; Grgic et al., 2020;
Ritti-Dias et al., 2011; Seo et al., 2012). DISCUSSION This suggests that a single familiarization trial is
sufficient to reduce the impact of a substantial learning effect (Bridgeman et al., 2016; do
Nascimento et al., 2017). Furthermore, the reliability values for both the 1RM and 5RM
(ICC = 0.97–0.98, CV = 2.1–2.7%) swing are towards the top of the range reported for
different exercises in a similar population (ICC = 0.64–0.99, CV = 0.5–7.8%) (Grgic et al.,
2020). An interesting finding from the current research is the similarity in reliability of the
1RM and 5RM kettlebell swing. The number of repetitions in RM tests has been shown to
have little difference in the reliability between a 1RM power clean (CV = 4.8%) and a 5RM
leg press (CV = 2.2–4.7%) (Gail & Künzell, 2014; Lawton, Cronin & McGuigan, 2014). Additionally, other work has shown reliability to be relatively stable across the number of
repetitions in an RM test (Gail & Künzell, 2014; Lattari et al., 2020; McCurdy et al., 2004;
Santos et al., 2019). The current results suggest that unless a 1RM value is specifically
required, a 5RM test may be a viable option as although the test is still maximal, the
absolute load will be lower and this may be important in certain populations (e.g., lower
training age, participants with lower kettlebell swing skill). A useful aspect of determining test-retest reliability is that it allows calculation of the
smallest worthwhile change (SWC) (Appleby, Cormack & Newton, 2019; Appleby, Newton
& Cormack, 2019). The SWC represents the smallest change in performance that is likely
to be of practical importance for athletic performance (Appleby, Cormack & Newton, 2019;
Appleby, Newton & Cormack, 2019). There are numerous methods that have been
proposed for calculating the SWC, including as a fraction (commonly 0.2) of the between
participant SD (Buchheit, 2016; Datson et al., 2021). Using this method, the TE/CV% of the
1RM and 5RM kettlebell swing is less than the SWC. As a result, for a change in
performance to be considered practically useful it must not only exceed the TE/CV% but in
this case also exceed the SWC (Appleby, Cormack & Newton, 2019; Appleby, Newton &
Cormack, 2019). The findings of this work are similar to those of previous work examining
a range of exercises such as the bench press, squat and arm curls where the TE/CV% was
less than the SWC (do Nascimento et al., 2017). Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 DISCUSSION An arguably more relevant aspect is the
signal-to-noise ratio (Crowcroft et al., 2017; Ryan, Kempton & Coutts, 2020). Whilst the
custom kettlebell device used in this research allowed small increases in load across
repetitions (2.5 kg), and therefore a likely relatively precise determination of 1RM and
5RM values, commercially available kettlebell increments are much larger (typically 8 kg or
up to 12 kg for loads >48 kg). In this case, the load increments in commercially available
kettlebells can be considered the “signal” and the CV% the “noise”. The fact that the load
increments often exceed the SWC means that any change in kettlebell 1RM or 5RM swing Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 7/13 performance observed using commercially available kettlebells, represents a practically
meaningful performance change. The use of a smallest increment of 2.5 kg for RM testing in the current study was based
on previous work (Sheppard et al., 2008), but has potentially resulted in a small
underestimation of the true 1RM and 5RM value. However, given the increments in
commercially available kettlebells typically exceed this amount, it does not impact the
finding that progression in RM performance from one kettlebell load to the next represents
a practically important change. Additionally, with a plate-loaded kettlebell as used in this study, the center of mass
(COM) changes with each load increment resulting in an overload from both the increased
mass and increased distance of the kettlebell COM from the fulcrum (Serway & Jewett,
1998). As both these factors will independently increase the required muscular force to
complete the kettlebell swing, they should be quantified. The kettlebell COM can be
calculated with the following equation where; m = mass and x = meters: COM = (m1x1 +
m2x2 + m3x3)/(m1 + m2 + m3) (Serway & Jewett, 1998). Further, commercially available
plate loaded kettlebells can be loaded with either a distal to proximal configuration, or a
proximal to distal configuration from the handle. Care should be taken to avoid the use of
interchangeable plate loading order. It is possible that RM loads were a function of changes
to both the mass and its distribution and therefore that the results may have been altered
with a kettlebell with different mass distribution. In contrast to a kettlebell swing with a plate-loaded kettlebell, 1RM barbell exercise
assessments may be more suitable for testing maximum strength. DISCUSSION Despite the 1RM being
considered the gold standard method of field-based testing for maximal strength, the
kettlebell swings relationship to maximal isometric strength is lower than other exercises
currently used for this purpose. For example, the barbell squat (r = 0.86–0.97) (Bazyler,
Beckham & Sato, 2015; Mcguigan et al., 2010), deadlift (r = 0.88) (De Witt et al., 2018),
snatch (r = 0.83), and the clean and jerk (r = 0.84) (Beckham et al., 2013), all have a
stronger relationship with isometric strength compared to the values reported for the 1RM
and 5RM kettlebell swing in the present study. Therefore, these other exercises offer better
validity for field-based tests to assess maximum strength. stronger relationship with isometric strength compared to the values reported for the 1RM
and 5RM kettlebell swing in the present study. Therefore, these other exercises offer better
validity for field-based tests to assess maximum strength. Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 Funding No funding was provided for this research. The funders had no role in study design, data
collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests Justin Keogh is an Academic Editor for PeerJ. The authors declare no other competing
interests associated with the publication of this article. Author Contributions James A. Ross conceived and designed the experiments, performed the experiments,
analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the
article, and approved the final draft. Justin W. L. Keogh conceived and designed the experiments, authored or reviewed drafts
of the article, and approved the final draft. Christian Lorenzen conceived and designed the experiments, analyzed the data,
authored or reviewed drafts of the article, and approved the final draft. ACKNOWLEDGEMENTS The authors would like to thank Associate Professor Stuart Cormack for his contribution
to the manuscript and the participants for their participation. CONCLUSIONS This research demonstrates that both the 1RM and 5RM kettlebell swings possess excellent
reliability. Critically, the SWC is less than the TE/CV% in both tests. Furthermore, both
these values are lower than commonly available kettlebell increments. Further research
using smaller increments during RM testing may allow more precise estimation of
reliability and the SWC. Practitioners can be confident that assessment of 1RM and 5RM
kettlebell performance following two familiarization sessions is not prone to large error. Due to the fact that commercially available kettlebell increments generally exceed the CV%
and SWC demonstrated in this research (i.e., signal > noise) (Appleby, Newton & Cormack,
2019), changes in performance based on an increase or decrease in 1RM or 5RM kettlebell
swing performance can be considered practically meaningful. Future research could
determine the effect of plate configuration upon biomechanical, physiological and Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 8/13 perceptual characteristics of the swing as well as the resulting adaptations such training
may reduce. Additionally, accurate knowledge of RM values may allow more precise
training prescription (e.g., % based loads), which could better elucidate the kettlebell
swing’s optimal training zones for strength and power adaptation. PRACTICAL APPLICATION Barbell exercises have a stronger relationship with isometric strength and are therefore a
better assessment of maximum strength. In contrast, the 1RM or 5RM swing is best used to
assess swing performance pre and post-training intervention. Practitioners may also wish
to consider plate loaded rather than fixed load kettlebells to allow more precise RM
determination. Further, if using a plate loaded kettlebell, plates of the same mass should
have the same width to ensure that the COM progressions are standardized. Plate loaded
kettlebells should be loaded in the same way, either distal to proximal from the handle or
proximal to distal from the handle. Finally, a 5RM kettlebell swing may represent a useful
alternative to a 1RM for lesser trained individuals. Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 Data Availability The following information was supplied regarding data availability: The raw data is available as a Supplemental File. The raw data is available as a Supplemental File. Supplemental Information Supplemental information for this article can be found online at http://dx.doi.org/10.7717/
peerj.14370#supplemental-information. Supplemental information for this article can be found online at http://dx.doi.org/10.7717/
peerj.14370#supplemental-information. Ross et al. (2022), PeerJ, DOI 10.7717/peerj.14370 Human Ethics The following information was supplied relating to ethical approvals (i.e., approving body
and any reference numbers): The following information was supplied relating to ethical approvals (i.e., approving body
and any reference numbers): Ethical approval was granted by the Australian Catholic University Human Research
Ethics Committee (2018-265E). Ethical approval was granted by the Australian Catholic University Human Research
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Genome-wide SNP analysis of three moose subspecies at the southern range limit in the contiguous United States
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Keywords Alces alces andersoni · Alces alces americana · Alces alces shirasi · Genotyping by sequencing (GBS) ·
Wildlife management · Single nucleotide polymorphism (SNP) Keywords Alces alces andersoni · Alces alces americana · Alces alces shirasi · Genotyping by sequencing (GBS) ·
Wildlife management · Single nucleotide polymorphism (SNP) Genome‑wide SNP analysis of three moose subspecies at the southern
range limit in the contiguous United States ason A. Ferrante1 · Chase H. Smith2 · Laura M. Thompson3 · Margaret E. Hunter1 Received: 14 April 2021 / Accepted: 16 September 2021 / Published online: 8 October 2021
This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2021 Received: 14 April 2021 / Accepted: 16 September 2021 / Published online: 8 October 2021
This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2021 * Jason A. Ferrante
jferrante@usgs.gov Conservation Genetics (2022) 23:109–121
https://doi.org/10.1007/s10592-021-01402-w Conservation Genetics (2022) 23:109–121
https://doi.org/10.1007/s10592-021-01402-w RESEARCH ARTICLE Jason A. Ferrante and Chase H. Smith contributed equally to this
work. 3
National Climate Adaptation Science Center, U.S. Geological
Survey, 12201 Sunrise Valley Drive, Mail Stop 516, Reston,
VA 20192, USA 1
Wetland and Aquatic Research Center, U.S. Geological
Survey, 7920 North West 71st Street, Gainesville, FL 32653,
USA Abstract Genome-wide evaluations of genetic diversity and population structure are important for informing management and conser-
vation of trailing-edge populations. North American moose (Alces alces) are declining along portions of the southern edge
of their range due to disease, species interactions, and marginal habitat, all of which may be exacerbated by climate change. We employed a genotyping by sequencing (GBS) approach in an effort to collect baseline information on the genetic vari-
ation of moose inhabiting the species’ southern range periphery in the contiguous United States. We identified 1920 single
nucleotide polymorphisms (SNPs) from 155 moose representing three subspecies from five states: A. a. americana (New
Hampshire), A. a. andersoni (Minnesota), and A. a. shirasi (Idaho, Montana, and Wyoming). Molecular analyses supported
three geographically isolated clusters, congruent with currently recognized subspecies. Additionally, while moderately low
genetic diversity was observed, there was little evidence of inbreeding. Results also indicated > 20% shared ancestry pro-
portions between A. a. shirasi samples from northern Montana and A. a. andersoni samples from Minnesota, indicating a
putative hybrid zone warranting further investigation. GBS has proven to be a simple and effective method for genome-wide
SNP discovery in moose and provides robust data for informing herd management and conservation priorities. With increas-
ing disease, predation, and climate related pressure on range edge moose populations in the United States, the use of SNP
data to identify gene flow between subspecies may prove a powerful tool for moose management and recovery, particularly
if hybrid moose are more able to adapt. Introduction Bowyer 2004). In North America, four subspecies have been
described based on biogeography and morphology (Peterson
1955; Hall 1981), with three of the subspecies’ boundaries
extending south into the contiguous United States (U.S.): A. a. shirasi (Shiras moose), A. a. andersoni (Western moose),
and A. a. americana (Eastern moose), and the fourth sub-
species, A. a. gigas (Alaskan moose), inhabiting Alaska and
northwestern Canada (Fig. 1). With an overall estimated
population of around one million in North America, A. alces
are not considered to be a species of concern by the United
States government (Timmermann and Rodgers 2017). There-
fore, management has predominantly been state or regionally
focused in an effort to maintain sustainable populations as
a local natural resource (Wattles and DeStefano 2011; Tim-
mermann and Rodgers 2017). Moose (Alces alces) are the sole extant members of the Alces
genus and are widely distributed among subarctic regions
of the northern hemisphere (Geist 1998; Hundertmark and Jason A. Ferrante and Chase H. Smith contributed equally to this
work. 1
Wetland and Aquatic Research Center, U.S. Geological
Survey, 7920 North West 71st Street, Gainesville, FL 32653,
USA 2
Department of Integrative Biology, University of Texas,
Austin, TX 78712, USA While some moose subpopulations are expanding, others
(notably along the southern periphery) are in steep decline
due to habitat loss and reduced forage quality (Timmermann 3
National Climate Adaptation Science Center, U.S. Geological
Survey, 12201 Sunrise Valley Drive, Mail Stop 516, Reston,
VA 20192, USA (0121 110 Conservation Genetics (2022) 23:109–121 and Rodgers 2017; Schrempp et al. 2019), increased preda-
tion (Mech and Fieberg 2014; Timmermann and Rodgers
2017), and increased prevalence of disease (Samuel 2007;
Lankester 2010; Ditmer et al. 2020; Ellingwood et al. 2020). In response, some local managers and state governments
have instituted regulations on hunting and other activities,
Fig. 1 Map depicting the range of moose (Alces alces) in the United
States and Canada with colors differentiating subspecies (adapted
from Jensen et al. 2018). Inset boxes indicate sampling sites for each
subspecies in our study. Northernmost samples in Montana (colored
orange) represent a subgroup of A. a. shirasi samples herein referred
to as NMT Fig. 1 Map depicting the range of moose (Alces alces) in the United
States and Canada with colors differentiating subspecies (adapted
from Jensen et al. 2018). Inset boxes indicate sampling sites for each subspecies in our study. Introduction Northernmost samples in Montana (colored
orange) represent a subgroup of A. a. shirasi samples herein referred
to as NMT and Rodgers 2017; Schrempp et al. 2019), increased preda-
tion (Mech and Fieberg 2014; Timmermann and Rodgers
2017), and increased prevalence of disease (Samuel 2007; Lankester 2010; Ditmer et al. 2020; Ellingwood et al. 2020). In response, some local managers and state governments
have instituted regulations on hunting and other activities, 1 3 3 Conservation Genetics (2022) 23:109–121 111 including an instance of ceasing permits for radio collar
telemetry-based population studies in Minnesota over con-
cerns that “unintended and unanticipated mortality of moose
occurred during the collaring process” (Minnesota, Execu-
tive Office of the Governor [Mark Dayton] 2015; Phillips
2021). As management authorities seek to reduce the rate of
decline, understanding the genetic makeup of these periph-
eral moose populations is key to prioritizing conservation
actions at the edge of the species’ range. including an instance of ceasing permits for radio collar
telemetry-based population studies in Minnesota over con-
cerns that “unintended and unanticipated mortality of moose
occurred during the collaring process” (Minnesota, Execu-
tive Office of the Governor [Mark Dayton] 2015; Phillips
2021). As management authorities seek to reduce the rate of
decline, understanding the genetic makeup of these periph-
eral moose populations is key to prioritizing conservation
actions at the edge of the species’ range. investigation of genetic diversity and structuring of popula-
tions (Davey et al. 2011; Kjeldsen et al. 2016; Roffler et al. 2016; Yang et al. 2016; Mérot et al. 2020). Despite its many benefits, the use of SNP methods to
investigate population genetic variation on moose has, to
date, focused primarily on a small number of populations in
Scandinavia (Nichols and Spong 2017; Blåhed et al. 2019). In the United States, Kalbfleisch et al. (2018) also analyzed
moose SNPs, but their study was limited to four individuals
across three subspecies (A. a. gigas, andersoni, and shirasi). For our study, we performed a SNP-based analysis to assess
current subspecies designations and inter-subspecies genetic
structure for the three recognized moose subspecies found
in the contiguous United States (A. a. americana, ander-
soni, and shirasi). Our opportunistic sampling targeted the
southernmost range of each subspecies, establishing base-
line SNP profiles for moose populations most susceptible to
environmental change. Sample collection We obtained a subset of moose tissue and blood samples
collected between 2009 and 2017 from Idaho (ID), Mon-
tana (MT), Minnesota (MN), New Hampshire (NH), and
Wyoming (WY) (Fig. 1). Samples included fresh blood
preserved in Tempus™ Blood RNA Tubes (ThermoFisher,
Waltham, MA), archived whole blood banked in – 80 °C
freezers, and tissue banked in – 20 °C freezers or desic-
cant (Ferrante et al. 2021). All samples had been collected
for research, during capture for radio collaring efforts or
health assessments, carcass recovery, or provided by hunt-
ers to the state agencies, and stored at the collaborator’s
respective facilities (Table 1). Samples from A. a. shirasi
were obtained from multiple states along almost the entire
latitudinal range of the subspecies in the United States. Sam-
ples from A. a. andersoni were obtained from the northwest
and northeast regions of Minnesota, where the subspecies
population in the United States is primarily located. Alces
a. americana had less representative sampling, consisting
solely of a collection effort encompassing a small region in
New Hampshire. Since the 1990’s, genetic studies of moose have relied
on cytogenic, mitochondrial DNA (mtDNA), and micro-
satellite markers to investigate genetic structure and sub-
species designations (Boeskorov 1997; Hundertmark et al. 2002a, 2003; Hundertmark and Bowyer 2004; DeCesare
et al. 2020), though genome-wide analyses are rare (e.g.,
Kalbfleisch et al. 2018). While mtDNA and microsatellites
have been useful for elucidating subspecies boundaries,
the use of genomic techniques provides increased capa-
bility for local-scale analysis that can better inform state
and regional conservation and management planning (e.g.,
Funk et al. 2012, 2019; Coates et al. 2018). In particular,
genotyping-by-sequencing (GBS) approaches (e.g., restric-
tion enzyme-based sequencing approaches) are increasingly
used in ecological and evolutionary research (Kjeldsen et al. 2016; Foote and Morin 2016; Ba et al. 2017; Cammen et al. 2018). These methods yield thousands of molecular mark-
ers to better resolve genetic differences, without the need of
a published genome, thus enabling a more comprehensive Introduction Herein, we describe our preliminary
findings on the genetic diversity and structuring of these
three subspecies with the goal of informing management
and recovery efforts at regional and state scales. Range edge populations are often characterized by
reduced effective population sizes (Piry et al. 1999; Bouzat
2010) and increased mating of related individuals (Neaves
et al. 2015). The subsequent reduction in genetic diversity
may reduce the fitness of individuals and the evolutionary
potential of a species, thereby increasing the probability
of population extinction (Bouzat 2010; Bijlsma and Loe-
schcke 2012; Mimura et al. 2017). Low genetic diversity is
a known trait among the North American moose populations
and likely reminiscent of a founder effect (Hundertmark and
Bowyer 2004). The North American moose population is
estimated to have been founded 11 to 14 Ka as a small popu-
lation of moose entered the Americas via the Beringian land
bridge (Guthrie 1995; Hundertmark et al. 2002b; Meiri et al. 2014). Subgroups of moose are hypothesized to then have
dispersed throughout North America via rare, long-distance
(leptokurtic) dispersal events (Hundertmark et al. 2003),
extending now into many northern U.S. states. Moose in
North America may have maintained inherently low levels of
genetic diversity since their founding, making it important to
take into account their demographic history when evaluating
levels of inbreeding and genetic health. Investigating cur-
rent genetic diversity among range edge moose populations
will provide a baseline for evaluating the impacts of climate
change on disease prevalence and habitat quality. Genetic analyses isolations from whole blood collected in Tempus tubes fol-
lowing Ferrante et al. (2018). DNA quantity was assessed
by spectrometry using an Epoch™ microplate spectropho-
tometer (BioTek, Winooski, VT) and integrity was ensured
by gel electrophoresis using a 0.8% agarose gel stained
with ethidium bromide. If there was no visible indication of
fragmentation, 1 µl of each sample was incubated in a Cut-
Smart® restriction enzyme buffer (New England BioLabs,
Ipswitch, MA) at 37 °C for 2 h and visualized by gel electro-
phoresis as a final quality control check prior to sequencing. We visualized the distribution of genetic variation among
SNPs in the moose subspecies using principal coordinates
analysis (PCoA) in the R package ‘adegenet’ v 2.1.1 (Jom-
bart 2008; Jombart and Ahmed 2011). We also used model-
based approaches to identify patterns of genetic structure
via the Bayesian clustering algorithm STRUCTURE v 2.3.4
(Pritchard et al. 2000) and the non-negative matrix factori-
zation algorithm TESS3 (Caye et al. 2016). For STRUCTU
RE, we ran 20 iterations where K, the number of clusters (or
populations), ranged from 1 to 7 (following lower rep test
runs to K = 10 classifying structure ~ K = 2 or 3), with an
initial burn-in period of 250,000 reps followed by 1 million
Markov chain Monte Carlo (MCMC) repetitions. The STRU
CTURE analysis was carried out on the USGS Yeti super-
computer (Falgout and Gordon 2015). We identified the
most appropriate K value in STRUCTURE HARVESTER,
which characterizes the data by the set of allele frequencies
at each SNP and maximizing Hardy–Weinberg equilibrium
within clusters (François and Durand 2010), by considering
the ΔK method (Evanno et al. 2005), the least negative mean
log likelihood (Ln P(D)), and the known geographic distri-
bution of the moose sampled (Pritchard et al. 2000; Earl and
vonHoldt 2012). Results were plotted using STRUCTURE
Plot v2.0 (Ramasamy et al. 2014) and it was deemed nota-
ble when samples had a q-value, or the proportion of SNPs
shared, was less than 0.8 within their cluster (> 20% shared
with another cluster). TESS3 was used to integrate collec-
tion localities with genotypic data. The program computes
ancestry proportions distributed over geographic space and
is useful in addition to STRUCTURE for identifying popu-
lation structure (Caye et al. 2016). TESS3 analyses were
performed in the R package ‘tess3r’ v 1.1.0 (Caye et al. Genetic analyses 2018) with the inclusion of geographic coordinates for each
sample, and we modeled K = 1–7. Cross-validation criterion
was used to select the most likely K and ancestry proportions
were mapped on an ancestry matrix. P i
i
F
l
l
l
d
i
h R
k DNA extraction and quality control DNA was extracted from tissue or whole blood using
DNeasy kits (Qiagen, Germantown, MD) following manu-
facturers protocols, or phenol–chloroform isoamyl alcohol 1 3 Conservation Genetics (2022) 23:109–121 112 Table 1 Moose (Alces alces
subspp) genotyping by
sequencing sample numbers (n)
and collection information
Samples were provided in various formats from collections obtained over a range of years
TEM: whole blood in Tempus™ Blood RNA Tubes
Subspecies
State
Sample type
Year(s) collected
n
A. a. shirasi
Idaho
Tissue (frozen), TEM
2017
10
Wyoming
TEM
2017
28
Montana
tissue (desiccated), TEM
2016–2017
30
A. a. andersoni
Minnesota
tissue (frozen), TEM
2009, 2011, 2013–2017
38
A. a. americana
New Hampshire
whole blood (frozen), TEM
2014–2017
53
Total
159 Samples were provided in various formats from collections obtained over a range of years
TEM: whole blood in Tempus™ Blood RNA Tubes Genotyping by sequencing between subspecies, states, and any other notable (> 20%
shared ancestry proportions) clusters or subclusters that
emerged following the STRUCTURE or TESS3 analyses. FST values of 0 to 0.05 and 0.05 to 0.15 were considered to
be of low to moderate differentiation, respectively, whereas
FST values > 0.15 were considered distinctly differentiated
(Hartl and Clark 1997). Estimates of genetic diversity were
conducted using the R package ‘diveRsity’ v 1.9.90 (Keenan
et al. 2013). Specifically, rarefied allelic richness (AR) was
used to compare the allelic richness among groups of sam-
ples of differing sizes, and observed heterozygosity (HO),
expected heterozygosity (HE), and the inbreeding coefficient
(FIS) were calculated. For AR and FIS, 999 bootstrap repli-
cates were used with a critical value of 0.05. region within A. a. shirasi displayed between 22 and 49%
shared ancestry proportions with the A. a. andersoni sam-
ples and were congruent to the observed group separated
from the A. a. shirasi cluster in the PCoA analysis (Fig. 2). No other moose samples displayed > 20% shared ancestry
with another cluster (Fig. 3B). TESS3 analyses supported a
K = 3 clustering (cross-validation plot; Fig. 4 inset), closely
aligning with the results from the PCoA and STRUCTURE
analyses. All samples were assigned within their designated
subspecies, and again, a high proportion of shared ancestry
(> 20%) was observed between the NMT samples and the A. a. andersoni moose (Fig. 4, dotted rectangle). The remaining
moose samples from MT were most similarly clustered with
the ID and WY samples. The FST values were low between A. a. andersoni and A. a. americana (FST = 0.129), while A. a. americana and A. a. shirasi were more strongly differentiated (FST = 0.265) show-
ing increasing differentiation with geographic distance west
to east (Table 3a). Among states, the largest differentiation
(FST = 0.302) was between WY and NH (Table 3b). Within
A. a. shirasi, the only subspecies sampled among multiple
states, moose samples displayed little differentiation (FST
range = 0.028 to 0.054) (Table 3b). As indicated by the
results from the STRUCTURE, PCoA, and TESS3, the NMT
A. a. shirasi subgroup was deemed notable and assessed as
an independent cluster from the remaining Montana moose
samples from the southern part of the state (hereafter SMT). Considered independently, A. a. Table 2 The number of moose
(Alces alces subspp.) samples
and SNPs remaining after each
of the sequenced filtering steps
for data quality control Genotyping by sequencing shirasi samples from NMT
displayed higher FST values between WY and ID than when
combined with all Montana moose samples, increasing from
FST = 0.054 to 0.123 with WY and FST = 0.028 to 0.09 with
ID. Both of these instances resulted in a change of desig-
nation from little to moderate differentiation between the
groups (Table 3b). The remaining SMT moose samples did
not display such a change in FST values, indicating low dif-
ferentiation between the subpopulations (FST < 0.05). Meas-
ures of genetic diversity among subspecies were similar,
with AR ranging from 1.801 to 1.901, and observed (HO) and
expected (HE) heterozygosity ranging from 0.229 to 0.268
and 0.253 to 0.268, respectively. The inbreeding coefficient
(FIS) was also low overall (range 0.010 to 0.099) among the Genotyping by sequencing The DArTseq GBS approach, developed and implemented
by Diversity Arrays Technologies (DArT; www.diversitya
rrays.com, Bruce, Australia) is useful for genetic mapping
and genome wide diversity analyses of species for which
no reference genome is available, as was the case with A. alces. Genotyping by sequencing was conducted by DArT
using ~ 600 ng of moose DNA. Samples were digested with
restriction enzymes PstI and SphI following Wenzl et al. (2004) prior to 75 base pair single end sequencing on an
Illumina HiSeq 2500 (San Diego, CA). Quality filtering and
SNP reporting was conducted using the proprietary software
DartSoft14 (Diversity Arrays Technology, Bruce, Australia)
pipeline.i Additional data filtering steps were performed in RStudio
(RStudio Team 2019) using the R package ‘dartR’’ v 1.1.11
(Gruber et al. 2018; Gruber and Georges 2019) following
similar methods as previous studies (Georges et al. 2018;
Smith et al. 2021). In the following order, SNPs or individu-
als were removed when: (1) the SNP marker reproducibility
(the proportion of alleles that give a repeatable result at a
locus) was below 1.00, (2) there was more than one SNP
per locus (i.e., the SNP with the highest degree of poly-
morphism was retained), (3) the locus had greater than 10%
missing data, (4) individuals were missing greater than 10%
of data, (5) the minimum allele frequency was less than 0.05,
and (6) the loci deviated from Hardy–Weinberg equilibrium
(HWE) (p = 0.0001). Pairwise FST values were calculated using the R pack-
age ‘StAMPP’ v 1.5.1 (Pembleton et al. 2013) and 999
bootstrap replicates to estimate population differentiation 1 3 3 113 Conservation Genetics (2022) 23:109–121 between subspecies, states, and any other notable (> 20%
shared ancestry proportions) clusters or subclusters that
emerged following the STRUCTURE or TESS3 analyses. FST values of 0 to 0.05 and 0.05 to 0.15 were considered to
be of low to moderate differentiation, respectively, whereas
FST values > 0.15 were considered distinctly differentiated
(Hartl and Clark 1997). Estimates of genetic diversity were
conducted using the R package ‘diveRsity’ v 1.9.90 (Keenan
et al. 2013). Specifically, rarefied allelic richness (AR) was
used to compare the allelic richness among groups of sam-
ples of differing sizes, and observed heterozygosity (HO),
expected heterozygosity (HE), and the inbreeding coefficient
(FIS) were calculated. For AR and FIS, 999 bootstrap repli-
cates were used with a critical value of 0.05. Results DArTseq analysis identified 9,780 SNPs across 159 moose
samples after quality filtering (Table 1). Our additional fil-
tering (Ferrante et al. 2021) yielded 1,920 SNPs from 155
moose (ID: n = 10; WY: n = 25; MT: n = 29; MN: n = 38;
NH: n = 53) which were found to be adequate for down-
stream analysis (Table 2). The PCoA depicted three clusters
congruent with the three taxonomically identified subspecies
(Fig. 2). The first two principal components (of 32 total)
explained 25% of the variation (20% and 5%, respectively),
with the 3rd and remaining components explaining 2.5% or
less each (not shown). Notably, a subgroup of moose sam-
ples (n = 12) identified as collected from northern MT (here-
after NMT) were observed separated from the main cluster
of A. a. shirasi moose in the direction of the A. a. andersoni
cluster (Fig. 1).i The STRUCTURE HARVESTER analysis best classified
the moose population structure as having K = 2 or K = 3 clus-
ters. The ΔK value peaked at K = 2 (Fig. 3A) with one clus-
ter representing A. a. shirasi and the other A. a. andersoni
and A. a. americana (Fig. 3B). The log probability of the
data plateaued at K = 3 (Fig. 3A) and groupings aligned with
existing subspecies designations and PCoA results (Figs. 2,
3B). In the K = 3 structure, all moose samples from the NMT 1 3
The number excluded by each filtering step is denoted in parentheses (-n)
Filtering step
Samples
SNPs
DArT quality control
159
9780
Filter by marker reproducibility (= 1.00)
159
6257 (− 3523)
Remove secondaries (i.e., 1 SNP per locus)
159
5985 (− 272)
Filter loci with significant missing data (< 90%)
159
5280 (− 705)
Filter out individuals with significant missing data (< 90%)
155 (− 4)
5111 (− 169)
Filter by MAF (< 0.05)
155
1920 (− 3191)
Filter out loci deviating from HWE
155
1920 114 Conservation Genetics (2022) 23:109–121 subspecies (Table 3a). When assessed by state, AR, HO, HE,
and FIS were moderately low throughout (Table 3c). support of previous microsatellite and mtDNA approaches. Although our sampling design was limited to opportunis-
Fig. 2 Principal coordinates
analysis (PCoA) results of
moose (Alces alces subspp.)
from the five states sampled
using 1,920 genomic SNP
loci. The first two coordinates
explained 25% of the varia-
tion. Three main clusters are
indicated. Discussion Our study produced the first SNP-based estimates of genetic
diversity for moose, which identified low baseline values for
all three subspecies. Genome-wide evaluations of genetic
variation are powerful approaches which can inform sub-
species designations, provide a basis for understanding pro-
cesses driving molecular diversification, and inform effec-
tive management and conservation strategies. Our GBS
approach produced nearly 2,000 SNP loci for the genomic
evaluation of U.S. moose without the need of a reference
genome, and provided an expansive genomic evaluation in Results Moose samples in
the dotted rectangle are from
the northern Montana (NMT)
sampling area
PCoA Axis 1 (20%)
PCoA Axis 2 (5%)
-10
-
5
5
0
-2
2
0
4
6
Idaho
Minnesota
Montana
New Hampshire
Wyoming
A. a. andersoni
A. a. americana
A. a. shirasi
NMT A. a. andersoni PCoA Axis 2 (5%)
0 A. a. shirasi A. a. americana subspecies (Table 3a). When assessed by state, AR, HO, HE,
and FIS were moderately low throughout (Table 3c). support of previous microsatellite and mtDNA approaches. Although our sampling design was limited to opportunis-
tic live sampling and archived samples, our initial survey
resulted in robust estimations of genetic diversity, patterns
of genetic structure, and a putative hybrid zone between two
moose subspecies—all of which will be useful in conserva-
tion and management planning, as discussed below. Fig. 2 Principal coordinates
analysis (PCoA) results of
moose (Alces alces subspp.)
from the five states sampled
using 1,920 genomic SNP
loci. The first two coordinates
explained 25% of the varia-
tion. Three main clusters are
indicated. Moose samples in
the dotted rectangle are from
the northern Montana (NMT)
sampling area Patterns of genetic variation in moose The primary goal of this study was to perform a SNP-based
analysis of North American moose at the southern range
edge of their distribution, where regional subpopulation
loss has been occurring. The results of our analyses sup-
port current subspecies designations (Figs. 2, 3, 4) and
added insight to existing diversity estimates. For instance, 1 3 3 115 Conservation Genetics (2022) 23:109–121 K=2
Idaho
Wyoming
Montana
Minnesota
New Hampshire
A. a. shirasi
A. a. andersoni
A. a. americana
A
B
K=3
Clu[West]
Clu[East]
Clu[Central]
NMT
Fig. 3 Results of the Bayesian clustering analysis using the program
STRUCTURE for 1,920 SNPs from moose (Alces alces subspp)
samples collected across five states (ID, WY, MT, MN, and NH). A
STRUCTURE HARVESTER graph displaying mean LnP(K) values
(right y-axis), and the Delta K values (left y-axis). The x-axis shows
the associated K cluster value. K = 2 and K = 3 indicated as likely
number of clusters in sample population. B STRUCTURE plots
showing the proportion of individual moose membership (y-axis) in
each cluster represented by a different color. Support was found for
both the K = 2 and K = 3 clustering results with the sample subspecies
designations aligning well, suggesting some level of genetic differ-
entiation observed at this resolution. A. a. shirasi samples with > 0.2
ancestry proportion shared with A. a. andersoni from area in northern
Montana labeled as NMT A A K=2
Idaho
Wyoming
Montana
Minnesota
New Hampshire
A. a. shirasi
A. a. andersoni
A. a. americana
B
K=3
Clu[West]
Clu[East]
Clu[Central]
NMT B Clu[Central] Clu[East] Fig. 3 Results of the Bayesian clustering analysis using the program
STRUCTURE for 1,920 SNPs from moose (Alces alces subspp)
samples collected across five states (ID, WY, MT, MN, and NH). A
STRUCTURE HARVESTER graph displaying mean LnP(K) values
(right y-axis), and the Delta K values (left y-axis). The x-axis shows
the associated K cluster value. K = 2 and K = 3 indicated as likely
number of clusters in sample population. B STRUCTURE plots showing the proportion of individual moose membership (y-axis) in
each cluster represented by a different color. Support was found for
both the K = 2 and K = 3 clustering results with the sample subspecies
designations aligning well, suggesting some level of genetic differ-
entiation observed at this resolution. A. a. shirasi samples with > 0.2
ancestry proportion shared with A. a. Patterns of genetic variation in moose The northern Montana (NMT)
samples are identified by the
dotted rectangle and correlate
with the PCoA and STRUCTU
RE subgroup observed. Idaho
(ID)
A. a. andersoni
A. a. americana
A. a. shirasi
Minnesota
New Hampshire
ID
Montana
Wyoming
0.0
0.8
0.6
0.4
0.2
1.0
Ancestry Proporons
NMT Conservation Genetics (2022) 23:109–121 116 Fig. 4 Moose (Alces alces sub-
spp.) ancestry matrix from the
TESS3 analysis showing K = 3
populations based on cross
validation score showing three
ancestral populations as most
likely estimate (plot inset). Indi-
vidual sample values are plotted
as vertical bars organized from
west to east based on their cap-
ture location. Colors represent
ancestry proportions of each
genetically differentiated cluster. The northern Montana (NMT)
samples are identified by the
dotted rectangle and correlate
with the PCoA and STRUCTU
RE subgroup observed. Idaho
(ID) A. a. andersoni
A. a. americana
A. a. shirasi
Minnesota
New Hampshire
ID
Montana
Wyoming
0.0
0.8
0.6
0.4
0.2
1.0
Ancestry Proporons
NMT 2006; Yıldırım et al. 2018; Teixeira and Huber 2021). In the
case of moose in the contiguous U.S., low genetic diversity
estimates are likely indicative to the demographic history
of North American moose or sampling design rather than
inbreeding. SNP-based estimates depicted lower heterozygo-
sity and allelic richness when compared to values calculated
using microsatellites in other portions of the North American
range, including Alaska (Schmidt et al. 2009; Wilson et al. 2015; DeCesare et al. 2020), western Canada (DeCesare
et al. 2020), and eastern Canada (Wilson et al. 2003). How-
ever, many of these studies focused on populations more lati-
tudinally centered in each subspecies’ range. Investigations
of genetic patterns in species populations, including in North
American ungulates, have generally found genetic diversity
to be lower at the margins and greater towards the center of
a species’ range (Eckert et al. 2008; Thompson et al. 2019). This is often due to isolation by distance or inbreeding as a
result of multiple biotic and abiotic factors (i.e. fragmented
habitats, low gene flow) working against them such as frag-
mented habitats, low gene flow, etc. (Kawecki 2008; Hun-
dertmark 2009). Estimates of genetic diversity in moose
appear to follow a similar trend; microsatellite-based esti-
mates from moose sampled closer to their range-edges were
more comparable to our SNP data estimates (DeCesare et al. 2020). Patterns of genetic variation in moose andersoni from area in northern
Montana labeled as NMT limits of each subspecies’ putative boundaries. Therefore,
future research targeting these areas would be beneficial for
delineating potential contact zones (see below discussion
on hybrid zones). However, within trailing edge subpopula-
tions, our SNP data did allow us to make some interesting
comparisons. our results show that genetic divergence (FST) is variable
between the three subspecies, with increasing differentiation
between subspecies correlating with geographic distance. Additionally, all subspecies displayed moderately low lev-
els of genetic diversity (i.e., AR, HO), very little to no evi-
dence of inbreeding (FIS), low loss of heterozygosity, and
little non-random mating (Table 3a). Our limited sampling
allowed for some intra-subspecies analysis, specifically for
A. a. shirasi, although our samples likely do not represent
the full SNP diversity within each subspecies. We therefore
caution over-interpretation to the level of entire subspecies
populations, as our study was limited by varied sampling
areas, and our ability to collect samples did not extend to the It is intriguing that the subspecies sampled within the
smallest geographic footprint (A. a. americana) had the low-
est inbreeding value (FIS = 0.01), suggesting that DNA was
collected from unrelated individuals. Northern regions of
NH are characterized by moderate moose densities asso-
ciated with favorable habitats (NH Fish and Game Com-
mission 2015) and it is, therefore, possible that sampling 1 3 116
Conservation Genetics (2022) 23:109–121
was less biased (i.e., less likely sampling of kin) than might
be expected given the small geographic footprint. Conse-
quently, our results suggest that relatively localized sub-
populations, including the one we surveyed in NH, may
not necessarily be at risk of negative effects like inbreeding
depression often seen in small, isolated subpopulations. It
should be noted that SNP assessments have been shown to
miss detecting inbreeding (FIS) that otherwise was appar-
ent via microsatellite analysis (Zimmerman et al. 2020). As
such, we feel our findings warrant more sampling from this
region to test these results. Additionally, this result may not
apply to all moose subpopulations in the contiguous United
States. For example, we see cases of geographically isolated
moose having higher inbreeding values as expected, such as
the A. a. Patterns of genetic variation in moose andersoni population on Michigan’s Isle Royale
(not included in this study) whose FIS values have doubled
between 1960–1965 (FIS = 0.08) and 2000–2005 (FIS = 0.16)
due to lack of gene flow from neighboring populations (Sat-
tler et al. 2017). A critical aspect of conservation biology is to establish
patterns of genetic diversity across geographic ranges of
species (Allendorf et al. 2013). Low genetic diversity can
affect the health of a species or population and may carry
a risk of extinction, but such outcomes may be overstated
as a certainty in conservation genetics (Holderegger et al. 2006; Yıldırım et al. 2018; Teixeira and Huber 2021). In the
case of moose in the contiguous U.S., low genetic diversity
estimates are likely indicative to the demographic history
of North American moose or sampling design rather than
inbreeding. SNP-based estimates depicted lower heterozygo-
sity and allelic richness when compared to values calculated
using microsatellites in other portions of the North American
range, including Alaska (Schmidt et al. 2009; Wilson et al. 2015; DeCesare et al. 2020), western Canada (DeCesare
et al. 2020), and eastern Canada (Wilson et al. 2003). How-
ever, many of these studies focused on populations more lati-
tudinally centered in each subspecies’ range. Investigations
of genetic patterns in species populations, including in North
American ungulates, have generally found genetic diversity
to be lower at the margins and greater towards the center of
a species’ range (Eckert et al. 2008; Thompson et al. 2019). This is often due to isolation by distance or inbreeding as a
result of multiple biotic and abiotic factors (i.e. fragmented
habitats, low gene flow) working against them such as frag-
mented habitats, low gene flow, etc. (Kawecki 2008; Hun-
dertmark 2009). Estimates of genetic diversity in moose
appear to follow a similar trend; microsatellite-based esti-
mates from moose sampled closer to their range-edges were
more comparable to our SNP data estimates (DeCesare et al. 2020). Alternatively, microsatellite-based methods may have
Fig. 4 Moose (Alces alces sub-
spp.) ancestry matrix from the
TESS3 analysis showing K = 3
populations based on cross
validation score showing three
ancestral populations as most
likely estimate (plot inset). Indi-
vidual sample values are plotted
as vertical bars organized from
west to east based on their cap-
ture location. Colors represent
ancestry proportions of each
genetically differentiated cluster. Patterns of genetic variation in moose Alternatively, microsatellite-based methods may have was less biased (i.e., less likely sampling of kin) than might
be expected given the small geographic footprint. Conse-
quently, our results suggest that relatively localized sub-
populations, including the one we surveyed in NH, may
not necessarily be at risk of negative effects like inbreeding
depression often seen in small, isolated subpopulations. It
should be noted that SNP assessments have been shown to
miss detecting inbreeding (FIS) that otherwise was appar-
ent via microsatellite analysis (Zimmerman et al. 2020). As
such, we feel our findings warrant more sampling from this
region to test these results. Additionally, this result may not
apply to all moose subpopulations in the contiguous United
States. For example, we see cases of geographically isolated
moose having higher inbreeding values as expected, such as
the A. a. andersoni population on Michigan’s Isle Royale
(not included in this study) whose FIS values have doubled
between 1960–1965 (FIS = 0.08) and 2000–2005 (FIS = 0.16)
due to lack of gene flow from neighboring populations (Sat-
tler et al. 2017). A critical aspect of conservation biology is to establish
patterns of genetic diversity across geographic ranges of
species (Allendorf et al. 2013). Low genetic diversity can
affect the health of a species or population and may carry
a risk of extinction, but such outcomes may be overstated
as a certainty in conservation genetics (Holderegger et al. 1 3 3 117 Conservation Genetics (2022) 23:109–121 groups, as well as including allelic richness (AR), observed heterozy-
gosity (HO), expected heterozygosity (HE), and inbreeding coefficient
(FIS) for (a) subspecies and (c) states Table 3 Population genetic statistics calculated using 1920 SNPs
from 155 moose (Alces alces subspp.) comparing proportional
genetic variance (FST) for (a) subspecies and (b) states and sub- a Value indicates distinct differentiation (> 0.15)
(a)
Subspecies
FST Amer
FST Ander
AR
HO
HE
FIS
A. a. americana
–
1.801
0.255
0.256
0.010
A. a. andersoni
0.129
–
1.901
0.268
0.286
0.058
A. a. Patterns of genetic variation in moose shirasi
0.265
0.154
1.833
0.229
0.253
0.099
Mean
1.845
0.259
0.265
0.056
(b)
State
FST NH
FST MN
FST WY
FST SMT
FST NMT
FST MT (All)
New Hampshire (NH)
–
0.242a
Minnesota (MN)
0.129
–
0.127
Wyoming (WY)
0.302a
0.193a
–
0.054
S Montana (SMT)
0.275a
0.160a
0.042
–
–
N Montana (NMT)
0.229a
0.111
0.123
0.079
–
–
Idaho (ID)
0.284a
0.166a
0.029
0.020
0.090
0.028
(c)
State
AR
HO
HE
FIS
New Hampshire
1.729
0.255
0.256
0.010
Minnesota
1.821
0.268
0.286
0.058
Montana
1.766
0.236
0.260
0.091
Wyoming
1.643
0.224
0.226
0.007
Idaho
1.661
0.220
0.226
0.023 pressure in the middle of the state locally extirpated moose,
dividing the moose into northern and southern subpopula-
tions that persisted through the twentieth century, even as
moose populations recovered. This human-mediated vicari-
ance event, paired with recent contact with populations of A. a. andersoni across central/western Canada, may explain the
genetic distinctiveness of NMT moose. Our findings align
with that study in the evidence for potential contact zones
between A. a. shirasi in NMT and A. a. andersoni in south
central Canada (DeCesare et al. 2020). SNP analysis of addi-
tional samples from central Canada and the Canadian Rock-
ies may provide the geographic coverage needed to resolve
the demographic history of these two populations, including
patterns of connectivity, interbreeding, and directionality of
gene flow. inflated estimates of genetic diversity throughout the range
of North American moose. The higher mutation rate and
multi-allelic nature of microsatellites could lead to overes-
timates of genetic diversity (Landegren et al. 1998; Zimmer-
man et al. 2020). Recent studies support the use of SNPs as
compared to microsatellite data for estimates of population-
level diversity due to their increased ability to differentiate
between clusters (Zimmerman et al. 2020). Further assess-
ment of moose throughout their subspecies distribution with
SNPs will be important to characterize genetic variation for
each subspecies across their range and to test whether micro-
satellite-based methods are overestimating genetic diversity. Conclusion The application of SNP analysis methods to targeted study
areas across moose species or subspecies ranges provides the
type of robust data needed to inform herd management and
conservation priorities. With increasing pressure on moose
populations in the United States, the ability to understand
genetic adaptation mechanisms is becoming even more
important. Specifically, the use of SNP data increases the
power of future research on moose demographics includ-
ing the leptokurtic (rare, long-distance) dispersal hypoth-
esis (Hundertmark et al. 2003; MacLeod et al. 2013) and
identification of gene regions under selection. We envision
further SNP analyses of this type including A. a. americana
samples from a broader geographic range, moose samples
from each subspecies range within southern Canada, and A. a. gigas samples from Alaska, thus allowing for a more fully
resolved demographic characterization of the current North
American moose population with specific benefit to local
subspecies populations. Contemporary moose populations are facing warming
average temperatures which affects their forage quality and
increases their exposure to diseases and parasitic infesta-
tions. This is evidenced by increased meningeal worm (Pare-
laphostrongylus tenuis) infestation through the expansion
of white-tailed deer into more northerly habitats, and sea-
sonal changes that support higher winter tick (Dermacentor
albipictus) survival (Murray et al. 2006; Lankester 2010;
Carstensen et al. 2019; Ellingwood et al. 2020). Genetic
variation plays a significant role in facilitating resilience
to a changing environment, and evolutionary adaptation is
an important process for improving an individual’s fitness
under such selective forces (Hendry et al. 2008). Many of the population genetic assessments on moose,
including our study, have focused on using neutral genetic
variation to resolve demography of populations over time
(e.g., phylogenetic relationships, mutation, gene flow). However, considering the anthropogenic and natural threats
moose currently face, future studies concerned with moose
genetic diversity might benefit from the investigation of
functional genetic diversity, or the measure of diversity of
potentially adaptive markers, by using RNA-seq and whole-
genome resequencing (WGR) methods (Brodie et al. 2021). This could be accomplished using the recently sequenced
European moose genome (Dussex et al. 2020), or opti-
mally, by first sequencing the nuclear genome of the North
American moose, and then looking to see which SNPs/genes
appear most impacted. Understanding both the genomic and
transcriptomic characteristics of successful moose subpopu-
lations, particularly in the face of increasing stressors, may
highlight which genes or genomic regions are influencing
their adaptive capacity. Future directions and implications on conservation Our analysis indicated that A. a. shirasi samples from NMT
share a notable proportion of SNPs with A. a. andersoni
samples (Figs. 2, 3B, 4), potentially indicating recent or
ongoing gene flow, divergence within A. a. shirasi, or clinal
variation (sensu Chafin et al. 2021) between the two subspe-
cies. DeCesare et al. (2020) recently characterized the last
century of moose populations in Montana using microsatel-
lite and mtDNA markers, and found evidence that hunting Due in part to their relatively large populations in North
America, moose have not been granted U.S. federal protec-
tion, even as a number of local populations have rapidly
decreased. Conservation efforts currently rely on state or
interstate management decisions, or on individual manage-
ment zones (e.g., parks, conservation areas, or protected 1 3 1 Conservation Genetics (2022) 23:109–121 118 moose subspecies may have similar implications for man-
agement in the context of a changing environment, particu-
larly if hybrid moose are more able to adapt. The integra-
tion of high-resolution genomic data from SNP analyses
into these studies can have a high value to conservation and
management planning for moose (Funk et al. 2019), as rein-
troduction or population augmentation can be restricted to
genotypes that maximize survivability in specific environ-
ments. Based on the results of such research, conservation
efforts may benefit from a shift to include special protection
for overlapping habitat in the future. lands). The results of our study provide baseline subspecies
genetic SNP diversity data for monitoring moose genetic
health at a local level. This may aid in long-term monitoring
of existing genetic diversity, assessing the effects of envi-
ronmental change on local moose populations, and possibly
inform human-mediated actions to improve genetic diver-
sity for future subpopulations. Essentially, states or regional
management entities can use this expanded genomic data to
coordinate management at the molecular level.i Our findings suggest edge subpopulations of A. a. ander-
soni and A. a. shirasi in northern Montana likely have had
opportunities for gene flow. The implications of overlapping
subspecies boundaries for local management could benefit
from multi-state management considerations for certain pop-
ulations. This includes consideration of movement corridors
between populations and decisions related to hunting limits
and land use. Such ecological passageways, where admixture
may occur, could be valuable for conservation, and poten-
tially support the adaptive capacity of moose in the face of
climate change (Rosvold et al. Future directions and implications on conservation 2013; Hendricks et al. 2019). Declarations Caye K, Jay F, Michel O, François O (2018) Fast inference of indi-
vidual admixture coefficients using geographic data. Ann Appl
Stat 12:586–608. https://doi.org/10.1214/17-AOAS1106i Conflict of interest: The authors declare that they have no competing
interests. Chafin TK, Zbinden ZD, Douglas MR et al (2021) Spatial population
genetics in heavily managed species: separating patterns of his-
torical translocation from contemporary gene flow in white-tailed
deer. Evol Appl 14:1673–1689. https://doi.org/10.1111/eva.13233 Research involving human and animal rights All samples collected
from live animals were obtained following ASM guidelines (Sikes
2016) under each collaborator’s respective IACUC approved methods
and permitting (Minnesota: IACUC UMN protocol #1601-33318A;
Montana: FWP12-2012; New Hampshire: IACUC UNH #130805;
Wyoming: IACUC #20150228MK00144-01, -02; ID: N/A). Research involving human and animal rights All samples collected
from live animals were obtained following ASM guidelines (Sikes
2016) under each collaborator’s respective IACUC approved methods
and permitting (Minnesota: IACUC UMN protocol #1601-33318A;
Montana: FWP12-2012; New Hampshire: IACUC UNH #130805;
Wyoming: IACUC #20150228MK00144-01, -02; ID: N/A). Coates DJ, Byrne M, Moritz C (2018) Genetic diversity and conserva-
tion units: dealing with the species-population continuum in the
age of genomics. Front Ecol Evol 6:165. https://doi.org/10.3389/
fevo.2018.00165 Open Access This article is licensed under a Creative Commons Attri-
bution 4.0 International License, which permits use, sharing, adapta-
tion, distribution and reproduction in any medium or format, as long
as you give appropriate credit to the original author(s) and the source,
provide a link to the Creative Commons licence, and indicate if changes
were made. The images or other third party material in this article are
included in the article's Creative Commons licence, unless indicated
otherwise in a credit line to the material. If material is not included in
the article's Creative Commons licence and your intended use is not
permitted by statutory regulation or exceeds the permitted use, you will
need to obtain permission directly from the copyright holder. To view a
copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Davey JW, Hohenlohe PA, Etter PD et al (2011) Genome-wide genetic
marker discovery and genotyping using next-generation sequenc-
ing. Nat Rev Genet 12:499–510. https://doi.org/10.1038/nrg3012 DeCesare NJ, Weckworth BV, Pilgrim KL et al (2020) Phylogeogra-
phy of moose in western North America. J Mammal 101:10–23. https://doi.org/10.1093/jmammal/gyz163 Ditmer MA, McGraw AM, Cornicelli L et al (2020) Using movement
ecology to investigate meningeal worm risk in moose, Alces alces. J Mammal 101:589–603. Conclusion Moreover, identifying gene flow
between subspecies and the potential resultant admixture of Acknowledgements This research was supported by the United States
Geological Survey National Climate Adaptation Science Center. This
study was dependent on samples provided by generous collaborators,
who we thank, from: (New Hampshire) P. Pekins and D. Ellingwood,
(Minnesota) T. Wolf, M. Carstensen, S. Moore, and E. Hildebrand,
(Idaho) M. Drew and C. Bleke, (Wyoming) M. Kauffman, A. May, and
B. Robb, and (Montana) N. DeCesare and M. Schwartz. We thank our
lab technicians E. Benzie, M. Giles and B. Caton for their invaluable
assistance, as well as the staff and technicians of our collaborators. We
are grateful to the USGS YETI team for their assistance. JAF thanks
MDF, MLT, EMF, the international Alces community and remembers
Dr. Vince Crichton. All authors are grateful for the thoughtful and thor-
ough review by the Journal’s reviewers who helped improve this work. Any use of trade, firm, or product names is for descriptive purposes
only and does not imply endorsement by the United States government. Author contributions Jason A. Ferrante and Chase H. Smith contrib-
uted equally to this manuscript. Laura M. Thompson, Jason A. Ferrante
and Margaret E. Hunter were responsible for the conception and design
of this study. Jason A. Ferrante and Chase H. Smith were responsible
for execution and analysis. The first draft of the manuscript was written
by Jason A. Ferrante and all authors contributed to the manuscript. All
authors read and approved the final manuscript. 1 3 3 119 Conservation Genetics (2022) 23:109–121 Funding This research was funded by the USGS National Climate
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functional and evolutionary diversity worldwide. Proc Natl
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1921849118 Data availability The data presented in this manuscript will be avail-
able from the USGS ScienceBase landing page (Ferrante et al. 2021):
https://www.sciencebase.gov/catalog/item/601864e4d34edf5c66f0
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servation units to promote recovery of two threatened freshwater 1 3 1 3
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3. Objectives Multiple genetic and environmental factors interact
resulting in loss of the upper motor neuron in the motor
cortex and the lower motor neurons cell bodies in the brain
stem and spinal cord [4, 5]. Pattern of onset could be spinal,
truncal, or bulbar. The clinical features of MND include
limb weakness, respiratory impairment, dysphagia, fatigue,
sleep disorders, pain, psychosocial distress, communication
deficits, cognitive impairment, and spasticity. Death occurs
secondary to respiratory failure 2 to 4 years after disease
onset on average; however survival of patients up to a decade
has been reported [6]. There is currently no cure for MND;
hence management is focused on symptomatic treatment,
rehabilitative care, and palliative care. The disease exerts a
huge psychological and economic burden on the patient and
caregivers. This review aims to objectively evaluate the role of the
multidisciplinary support care available to patients with
MND, the evidence basis for intervention modalities, and
highlight areas for future research. The benefit(s) of inter-
vention measures are assessed on their impact on outcome
measures such as survival, quality of life (QOL), decreased
hospitalization, improved 3 disability, and cost effectiveness. orrespondence should be addressed to U. E. Williams; williamsuduak@yahoo.co.uk Correspondence should be addressed to U. E. Williams; williamsuduak@yahoo.co.uk Received 20 June 2014; Revised 29 September 2014; Accepted 28 October 2014; Published 18 November 2014
Academic Editor: Anabela C Pinto Received 20 June 2014; Revised 29 September 2014; Accepted 28 October 2014; Published 18 November 201 Academic Editor: Anabela C. Pinto Copyright © 2014 U. E. Williams et al. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Motor neuron disease is a neurodegenerative disease characterized by loss of upper motor neuron in the motor cortex and lower
motor neurons in the brain stem and spinal cord. Death occurs 2–4 years after the onset of the disease. A complex interplay of
cellular processes such as mitochondrial dysfunction, oxidative stress, excitotoxicity, and impaired axonal transport are proposed
pathogenetic processes underlying neuronal cell loss. Currently evidence exists for the use of riluzole as a disease modifying drug;
multidisciplinary team care approach to patient management; noninvasive ventilation for respiratory management; botulinum toxin
B for sialorrhoea treatment; palliative care throughout the course of the disease; and Modafinil use for fatigue treatment. Further
research is needed in management of dysphagia, bronchial secretion, pseudobulbar affect, spasticity, cramps, insomnia, cognitive
impairment, and communication in motor neuron disease. 2. Review Strategy Evidence for this review was obtained from a search of the
Cochrane data base, PUBMED, guidelines of National Insti-
tute for Clinical Excellence (NICE), American Academy of
Neurology (AAN), and European Federation of Neurological
Societies (EFNS); and peer-reviewed journal articles. MND
diagnosis is based on the El Escorial diagnostic criteria [4, 5]. Motor neuron disease (MND) also referred to as amyotrophic
lateral sclerosis (ALS) is a fatal neurodegenerative condition
with an annual incidence of about 1.5 per 100,000 [1] and a
United Kingdom (UK) prevalence of 4–6/100,000 [2]. There
is a slight male preponderance with a male to female ratio of
3 : 2. It could occur at any age but the peak age of occurrence
is between 50 and 75 years [3]. Hindawi Publishing Corporation
Journal of Neurodegenerative Diseases
Volume 2014, Article ID 435164, 10 pages
http://dx.doi.org/10.1155/2014/435164 Hindawi Publishing Corporation
Journal of Neurodegenerative Diseases
Volume 2014, Article ID 435164, 10 pages
http://dx.doi.org/10.1155/2014/435164 Hindawi Publishing Corporation
Journal of Neurodegenerative Diseases
Volume 2014, Article ID 435164, 10 pages
http://dx.doi.org/10.1155/2014/435164 U. E. Williams, E. E. Philip-Ephraim, and S. K. Oparah Internal Medicine Department, University of Calabar, Calabar, Cross River State 540242, Nigeria Journal of Neurodegenerative Diseases The low riluzole and noninvasive
ventilation (NIV) use has been suggested as the reason why
there was no difference in survival observed in this study
[12]. Another Italian study [17], reviewing 221 participants in
a MDC setting, noted an improved median survival (𝑃=
0.008), decreased hospitalization (1.2 admission frequency
versus 3.3, 𝑃= 0.003), and decreased duration of hospital
stay (5.8 versus 12.4 days, 𝑃= 0.001) in the MDC cohort. Riluzole is the only registered DMT for MND which
slows the disease progression but does not stop the under-
lying disorder. The mechanism of action of riluzole involves
blocking of presynaptic glutamate release [24]. Four trials
[25–28] provided the evidence base for riluzole as a DMT. The first controlled trial involving riluzole [25] reported a
modest increase survival among treated patients compared
to controls receiving placebo. The same group undertook a
subsequent study [26] to address some of the issues raised in
the pilot study and confirmed that riluzole is well tolerated
and it also extends survival of MND patients. The meta-
analysis of these studies showed that irrespective of the
patient selection, 100 mg prolongs median survival in people
with MND by 2-3 months [29]. Minor reversible adverse
reactions reported were nausea, asthenia, fatigue, and an
increase in liver enzymes. A recent population-based study
[16] found a 6-month overall survival benefit that was signif-
icant in bulbar onset and elderly patients, but not in patients
with a limb-onset disease. No additional benefits were noted
when coadministered with add-on such as Vitamin E [30] or
Gabapentin [31]. A group [18] retrospectively reviewed hospital notes of
162 patients seen between 1998 and 2002 in GNC and 255
others managed under MDC care between 2006 and 2010
in a tertiary hospital. Median survival from diagnosis was
19 months for MDC and 11 months for GNC (Hazard ratio
0.51, 95% confidence interval 0.41–0.64). They also analyzed
the relationship between MDC and survival independent of
riluzole, NIV, and PEG use. Although this study selected
patients from multiple neurologists in the region, a rigorous
methodology was used to ensure proper patient selection
and matching. Other factors that are being suggested as
contributing to an improved outcome in MDC setting include
better symptomatic support, access to aids, and prompt
treatment of respiratory challenges [14]. Journal of Neurodegenerative Diseases 2 Table 1: Genetics of MND. Type of MND
Genetic mutation
Familial MND
(i) SOD1 gene [100]. (ii) TDP-43 [101–103]. (iii) Alsin (ALS2) [104, 105]. (iv) Senataxin (ALS4) [106]. (v) Vesicle associated membrane-protein
(VAPB, ALS8) [107]. (vi) Angiogenin [108, 109]. (vii) Mutation in the p150 subunit of dynactin
(DCTN1) [110, 111]. Sporadic MND
Genetic mutations linked to greater
susceptibility to sporadic MND include
(1) apolipoprotein E4, [112]
(2) decreased expression of excitatory amino
acid transporter-2 protein [113, 114],
(3) alterations in the vascular endothelial
growth factor (VEGF) gene [115]. Table 1: Genetics of MND. such as multiple sclerosis [7], stroke [8], acquired brain injury
[9], and MND. MDC is defined as any care delivered by two
or more disciplines [10], involving a neurologist and other
allied disciplines such MND nurse, chest physiologist, and
occupational therapist. Other personnel needed as part of the
MDC team for MND care includes occupational therapists,
physiotherapists, social workers, counselors, speech and lan-
guage therapist, and religious leaders. Care is administered
24 hours daily in a hospital or on outpatient basis or in the
patients’ home or community, but effort must be effectively
coordinated to avoid overlapping or missing care due to the
large number of care providers involved in the management
of the patient and their family. MDC is important in enabling
care specialist to undertake proper assessment of patients and
addressing the concerns of patients and family [11, 12]. An Irish prospective population-based cohort study [13]
compared 344 patients in MDC to patients in general
neurology care (GNC) and found 7.5-month longer survival
in the MDC cohort (𝑃< 0.004). Another cross-sectional
study involving 208 participants with MND [14] observed an
improved QOL in patients with MND who attended MND
clinic 6–12 weekly compared to participants who attended a
6-monthly GNC. In a subsequent report [15] observed no dif-
ference in healthcare cost between MDC and GNC settings. interplay between complex cellular processes acting syner-
gistically is accumulating [21, 22]. Mutation in super oxide
dismutase (SOD1) and TAR-DNA binding protein (TDP-
43) among others are strong genetic risk factors [21, 23]. A
summary of the genetic and pathophysiologic processes in
MND is summarized in Tables 1 and 2. g
In an Italian study involving 126 ALS patients [16], no
difference in the median survival time between MDC care
and a GNC cohort was reported (17.6 months versus 18
months; 𝑃
=
0.76). 4. Evidence for Multidisciplinary Care (MDC)
Strategies and Modalities 4.1. Care Setting. MDC approach is the main stay for the
management of patients with chronic neurological conditions Journal of Neurodegenerative Diseases 2 Journal of Neurodegenerative Diseases Both AAN and
EFNS recommend MDC care setting for patients with MND,
with the current EFNS guidelines recognizing the benefit
of MDC approach in improving survival, reducing medical
complications, and improving the quality of life of patients
and their caregivers [19, 20]. Recombinant human insulin-like growth factor-1 (rhIGF-
1) has been proposed as a MDT in MND following its ability
to promote spinal motor neuron survival after excitatory
amino acid induced death in animal models [32, 33]. A
Cochrane review [34] of the benefit of rhIGF-1 on disease
progression using 3 studies involving 799 MND patients
observed low quality evidence of improved QOL scores at 9
months, with no impact on survival. A meta-analysis of 2 well
randomized trials using ciliary neurotrophic factor (CNTF)
as a DMT in 1300 MND patients showed no significant 4.2. Neuroprotective Treatment and Disease-Modifying Ther-
apy (DMT). The exact molecular pathway leading to the loss
of motor neurons in MND remains unclear, but evidence of 3 Journal of Neurodegenerative Diseases Journal of Neurodegenerative Diseases 3 Table 2: Pathophysiological processes in MND. Table 2: Pathophysiological processes in MND. Pathophysiologic process
Comments
Excitotoxicity
Excessive postsynaptic glutamate induced stimulation of glutamate receptors such as NMDA & AMPA
→massive calcium influx →nitric acid formation and neuronal death [21, 116]. Oxidative stress
Fibroblast culture from MND patients shows increased sensitivity to oxidative damage. Accumulation of
free oxygen species →cell death. SOD1 is an antioxidant enzyme [116]. Mitochondrial defect
Abnormalities of mitochondrial morphology and biochemistry have been reported in sporadic MND
patients, in SOD1 transgenic mice, and in cellular models [117, 118]. Impaired axonal transport
The relatively long length of motor neuron depends on effective transport systems. Evidence of
abnormalities in this transport system has been reported in transgenic mice [119–121]. Neurofilament aggregation
Abnormal accumulation of neurofilament commonly occurs in many neurodegenerative diseases
including MND [122, 123]. Protein aggregation
Intracellular inclusions have been observed in MND. The evidence is still unclear if these proteins are
toxic or beneficial to the cell [21, 22]. Inflammatory dysfunction
Evidence suggests the possibility of an inflammatory process [23]. Deficits in neurotrophic
factors and dysfunction of
signaling pathway
Deficits in levels of neurotrophic factors, e.g., IGF-1, have been reported in MND [124–126]. Apoptosis
The final process in MND leading to neuronal death is said to closely resemble apoptosis, and markers of
apoptosis have been detected in the later stages of the disease and animal models [127–129]. Journal of Neurodegenerative Diseases difference between it and placebo, unlike findings reported
in animal models which were favorable [35].h In a subsequent randomized control trial (RCT) [46]
involving 22 MND patients on NIV and 19 on standard
care, a 48-day longer mean survival in the NIV group was
observed compared to the standard care group (𝑃= 0.0062)
at 12 months. In the subgroup with good to moderate bulbar
function survival was 205 days longer (𝑃= 0.0059). The
strength of this study was the computer-based randomized
allocation of patients to respiratory support and the even
matching of patients and controls in terms of demographic
characteristic and functional ability. This study has docu-
mented convincingly that NIV prolongs median survival. A
review by NICE has further confirmed the cost effectiveness
of NIV used for MND patients [44]. NIV improves gas
exchange, alleviates symptoms of carbon dioxide retention,
and improved QOL [47]. The exact mechanism of action
of NIV is unknown, but it may be related to the reversing
of chronic respiratory fatigue, reversing of hypercapnia,
resolution of atelectasis, and decreasing of the rate of decline
of FVC [48]. Claustrophobia, anxiety, excessive salivation,
nasal bridge soreness, and abdominal bloating are some of
the problems associated with NIV use in MND patients [41].hf The proposed involvement of free radical accumulation
and oxidative stress in MND has informed the trial of
antioxidants in MND. A meta-analysis of 10 studies involving
1015 MND patients [36] reported weak evidence for the
efficacy of antioxidants in MND. Some of the antioxidants
which have had positive effect in animal studies are vitamins
C and E [37], selegiline [38], N-acetylcysteine [39], and
dehydroepiandrosterone [40]. 4.3. Symptomatic Management It is not encouraged in the USA or
Europe but it is the most commonly used respiratory support
in Japan [52, 53]. risk of aspiration due to sedation and its being unsuitable for
patients with moderate to severe respiratory dysfunction are
some of the drawbacks of PEG [48, 72]. RIG requires local anaesthesia for its insertion; thus it
is observed to have higher success and lower complication
rate [68]. It can be used even when FVC is <50%. The
disadvantage of RIG is that it is not as securely fixed as
PEG and the small tube used can easily be blocked. PIG
is a new fluoroscopic technique that has a better long-term
clinical outcome in terms of success and complication rates. It requires only local anaesthesia or minimal sedation, and the
tube rarely blocks or migrates [70].h There is no evidence in literature regarding which inser-
tion method is superior in any given circumstance [73]. The
need for robust randomized prospective trials is encouraged
by both AAN and EFNS [19, 64]. Three recent studies [74–76]
could not demonstrate any significant difference in survival
between PEG and RIG. Another study observed significant
difference (𝑃= 0.004) between survival in PEG and RIG
patients in a subgroup with respiratory failure [68]. The
median survival after gastrostomy was 140% higher in the
RIG group compared to PEG patients. It is believed that
suggestions of PEG being better are just tentative conclusions. [77]. Respiration can also be assisted using an electrical stim-
ulation of the diaphragm to produce contractions (diaphrag-
matic pacing). It is a procedure originally meant for patients
with cervical spine injury [54], but still experimental in
MND. Four electrodes are placed on the motor roots of the
phrenic nerves on the abdominal surface of the diaphragm. Thus it is only effective if the diaphragm still retains some
innervation [41]. The diaphragmatic pacing in patients with
respiratory muscle weakness due to motor neurone diseases-
tudy (DIPALS) is an ongoing RCT assessing the efficacy of
diaphragmatic pacing among MND patients in some UK
hospitals. Home parenteral nutrition (HPN) using a central venous
catheter is considered as an alternative to long-term nutri-
tional support for MND patients with dysphagia when
gastrostomy is contraindicated due to severe respiratory
distress. Though it is more expensive than gastrostomy, recent
evidence that HPN can be well tolerated and can improve
nutritional status is patient with MND [78]. 4.3. Symptomatic Management 4.3.1. Respiratory Management. Respiratory impairment is
the leading cause of death in MND. Denervation weakness of
respiratory muscles results in ineffective cough, retention of
secretion, and hypoventilation, and it is an important deter-
minant of QOL [41–43]. Its proper management can improve
survival and QOL [44]. Onset of respiratory impairment is
marked by sleep disordered breathing (SDB) which results
in early morning headaches, nonrefreshing sleep, daytime
somnolence, dyspnea, orthopnea, poor concentration, and
fatigue [41]. Assisted ventilation in MND can be provided
using invasive technique via tracheostomy or NIV using face
or nasal mask. The effectiveness of NIV can be enhanced by the use
of telemonitoring of NIV. Pinto et al. [49] evaluated home
telemonitoring of NIV in ALS patients and observed that
there was improved survival and lower office and emergency
room visit or admissions among patients who had telemoni-
toring compared with the control group which who had their
compliance and ventilator parameter settings assessed only
during clinic visits. They concluded that telemonitoring help
reduce health care cost and improve survival and functional
status. In a study involving 22 subjects with MND [45], a 2-
monthly assessment of QOL using Short Form Health Survey
(SF-36), chronic respiratory disease questionnaire, sleeps
apnoea quality of life index, and respiratory function and a
4-monthly polysomnography showed improvement in sleep
related problems and mental health that was maintained for
252 to 458 days. Overall survival was significant between
NIV patients and those on standard care. Moderate to severe
bulbar weakness was associated with a lower improvement in
QOL. Invasive ventilation or tracheostomy ventilation (TV)
can also be used to deliver air into the lungs and to clear
secretions. It is used in patients with severe bulbar dys-
function who cannot tolerate NIV or in patients previously 4 Journal of Neurodegenerative Diseases using NIV whose respiratory function has deteriorated to a
point where NIV is not tolerated [41]. A retrospective chart
review combined with prospective evaluation of QOL and
degree of depression [50] obtained a survival rate of 65% by
1 year and 45% by 2 years after tracheostomy. Survival was
significantly shorter in patients older than 60 years with a
hazard ratio of dying of 2.1 (95% confidence interval, 1.1–3.9). While TV allows for suction of secretion and avoids facemask
and claustrophobia, it predisposes to recurrent infection, tra-
cheostomy site infection, bleeding, and tracheaoesophageal
fistula formation [51]. 4.3. Symptomatic Management Hypercaloric
enteric nutrition has been proposed as a factor that can
improve survival because mild obesity is associated with
improved survival [79]. Wills et al. [79] evaluated the safety
and tolerability of high carbohydrate hypercaloric diet in
patients with MND receiving enteric nutrition and observed
that when compared to patients receiving isocaloric tube-fed
diet or high fat hypercaloric diet, there was lower adverse
effect or serious adverse event in the high-carbohydrate
hypercaloric group. They were of the view that high carbo-
hydrate hypercaloric enteric nutrition is safe and tolerable in
patients with MND. Apart from hypoventilation, respiratory weakness also
impairs cough [55]. Insufficient cough results in recurrent
chest infection, which is the leading cause of hospitalization
in MND [56]. The strength of the patients’ cough is assessed
with a peak flow meter and reported as suboptimal if the peak
cough-flow (PCF) is less than 270/min [57]. Cough can be
augmented using intensive physiotherapy and manoeuvres
like tussive squeeze and mechanical in-exsufflator (MI-E). Evidence for MI-E is weak, but it has been suggested that it
could be effective in MND patients for cough management
[58, 59]. 4.3.2. Nutritional Management. Dysphagia may occur in
MND due to loss of coordination, weakness of muscles
of mastication, weakness of tongue, and impaired swallow-
ing. This can be complicated by weight loss, distressing
choking, prolonged effortful meal times, frequent aspiration,
and increase risk of chest infection [60–62]. Malnutrition
positively correlates with a shortened survival rate and is
a poor prognostic factor [63]. Management of dysphagia at
the early stage involves changes to food texture and teaching
of swallowing techniques. In the later stages feeding can be
through nasogastric tube (NGT) or gastrostomy insertion
[64–66]. Current principles of care for dysphagia in MND
are based on consensus and expert opinion rather than
randomized controlled trials [67]. 4.3.3. Other Symptomatic Management Sialorrhoea. Excessive salivation is common in MND as bul-
bar dysfunction worsens and can be embarrassing or result
in aspiration. Amitriptyline, Atropine, Botulin toxin type-B
(BTX-B), and external irradiation of the salivary gland have
all been tried in the control of sialorrhoea. A double-blind
control trial BTX-B injected into the parotid and mandibular
gland of 20 patients with refractory sialorrhoea achieved 82%
reported improvement compared to 38% among those who
received placebo at 2 months (𝑃< 0.05) [80]. Costa et
al. [81] also evaluated the efficacy and safety of BTX-B in
the treatment of sialorrhoea in patients with a bulbar onset
MND in an open-labeled prospective study that involved
the injection of BTX-B into the parotid and submandibular
glands. They observed that most patients reported a better Guidelines of both the AAN and EFNS recommend the
use of gastrostomy in MND [19, 20]. Methods of gastros-
tomy insertion include percutaneous endoscopic gastros-
tomy (PEG), radiologically inserted gastrostomy (RIG), and
Per-oral image-guided gastrostomy (PIG) [68, 69]. In PEG
which is the most commonly used method a wide bore-tube
is fixed under endoscopic guidance after sedation to reduce
the risk of migration and blockade [70, 71]. The increased 5 Journal of Neurodegenerative Diseases of weakness. Physical therapy can be combined with one or
more of the antispasticity drugs [92]. quality of life while on treatment and a mean reduction of
symptom severity of 70%. The most commonly reported side
effects include viscous saliva, local pain, chewing weakness,
and respiratory infection. Insomnia and Fatigue. Insomnia is common in the final
stages of MND probably due to cramps, pain, and respiratory
impairment [93]. Amitriptyline and Zolpidem are some of the
medications used in practice without being tested. Fatigue is
potentially debilitating and may be of central or peripheral
origin. An open-labeled trial confirmed the effectiveness of
Modafinil in the treatment of fatigue in MND [94, 95]. Bronchial Secretion. Bulbar impairment results in poor clear-
ance of tenacious sputum. Mucus accumulation is a poor
prognostic factor in patients on NIV [82]. No RCT exists for
any of the treatment approaches in MND. EFNS recommends
the use of mucolytics such as N-acetylcysteine when there is
sufficient cough flow. Cognitive Impairment. MND is associated with a frontotem-
poral type of dementia and it is associated with a negative
impact on survival [96]. Cognitive impairment has been
demonstrated in 20–50% of patients with MND. 4.3.3. Other Symptomatic Management A number
of screening instruments are available for assessing cognitive
impairment in MND, but EFNS recommends the use of tools
that can assesses verbal fluency as a major component of any
test instrument [20]. Pseudobulbar Affect. This is observed in up to 50% of MND
patients [83]. Yawning, weeping, and laughing are the char-
acteristic presentation. Pseudobulbar affect has a negative
impact on QOL. Small placebo controlled trials and case
series have observed the effectiveness of selective serotonin
reuptake inhibitors (SSRI) and tricyclic antidepressants in
controlling this symptom [84, 85]. Pioro et al. [86] evalu-
ated the treatment of pseudobulbar affect in patients with
multiple sclerosis and MND in a randomized-controlled trial
using 30/10 mg and 20/10 mg Dextromethorphan plus ultra-
low-dose quinidine. They reported that dextromethorphan
plus ultra-low-dose quinidine is effective in reducing the
frequency and severity of symptoms and improving patients’
quality of life especially at the dose of 30/10 mg combi-
nation when compared with placebo. EFNS recommends
Citalopram (SSRI) and Amitriptyline (TCA) for treatment of
troublesome cases of pseudobulbar affect [20]. A fixed dose
combination of dextromethorphan/quinidine (30 mg/30 mg)
is the recommended treatment option of AAN [87]. Communication. It is important for effective social interac-
tions. Subtle changes may be seen as word finding difficulties,
spelling difficulty, and decreased verbal output. Language
impairment results in difficulties in clinical management
and decreases QOL of patients and caregivers [97]. EFNS
recommends 3–6 monthly assessment; full neuropsychology
test; and use of communication aids like computerized speech
synthesizer [20]. Palliative Care. This is a holistic management of patients with
terminal disease aimed at optimizing the QOL of patients
and their caregivers. Palliative care in MND should ideally
start at the point of diagnosis and continue throughout the
entire history of the disease. It is needed at the different stages
of the disease such as at the time of diagnosis to address
issues relating to breaking of the bad news to patient and their
caregivers; during crises points for example, introduction of
NIV or PEG; and terminal stage when the patients’ condition
deteriorates. The application of the palliative care techniques
should be organized to meet the need of individual patients as
the reaction of patients and their families to the evolution of
the disease varies between patients. Journal of Neurodegenerative Diseases 6 6 talking about death and knowing that one’s family is prepared
for their death among others are issues which are important
for palliative care. They also largely accepted pain and symp-
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Neurology, vol. 65, no. 8, pp. 1264–1267, 2005. MND is a fatal syndrome with short disease course and low
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no. 1, pp. 27–34, 2009. 5. Areas Needing Further Research in MND Further research is needed in examining referral bias to
MDC and other factors, which may influence effectiveness of
MDC clinic outcomes such as frequency of visit. Biomarker
for early diagnosis should be evaluated as this aids in
research into DMT. Better evidence is still needed in the best
parenteral feeding modality and the most optimal time of
initiating/withdrawing it; impact of cough-assisting devices;
nutritional management techniques on the QOL; evaluation
of language dysfunction and its management treatment; the
cost effectiveness of different treatment modalities; approach
to the management of terminal care; and evidence for
advance directives, impact of disease, and effect of disease on
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“Effect of a multidisciplinary amyotrophic lateral sclerosis
(ALS) clinic on ALS survival: a population based study, 1996–
2000,” Journal of Neurology, Neurosurgery and Psychiatry, vol. 74, no. 9, pp. 1258–1261, 2003. Conflict of Interests [16] S. Zoccolella, E. Beghi, G. Palagano et al., “Riluzole and amy-
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of care such as physical (symptom control); psychological
(effect of disease on patients); social (impact of disease on
family and care-givers); and spiritual (questions bordering
on meaning of life and the fear of dying). The approach
should integrate both clinic and community-based care from
onset of disease and continue even after the patient has died
[11, 98]. EFNS guidelines [20] recommend early palliative
care referral with discussion covering aspects of end-of-life,
advance directive, and naming of health-care proxy. Cramps. It is usually troublesome particularly at night. A
RCT failed to support the efficacy of tetracannabinoid in
treating moderate to severe cramp [88]. A small open-
labeled pilot study confirmed that levetiracetam is useful
for treatment of cramps in MND patients [89]. Modalities
such as massage, physical exercise, hydrotherapy, heated
pools, and drugs like carbamazepine, diazepam, phenytoin,
and verapamil have all been tried without any conclusive
evidence. EFNS recommends levetiracetam, quinine sulfate,
and physical therapies for management of cramp in MND. Spasticity. Physical therapy is the main treatment modality for
spasticity that has its usefulness established from random-
ized controlled trial in literature [20, 90]. Physical therapy
methods in use include therapeutic exercise, stretching,
positioning, casting, and biofeedback. Other interventions
with no controlled trial evidence include heat/cold ther-
apy, hydrotherapy, ultrasound, electrical stimulation, and
chemodenervation and rarely surgery can be used [20]. Intrathecal Baclofen is the drug of choice in intractable cases
[20, 91]. Drugs such as Dantrolene, Tetrazepam, and Tizani-
dine have not been tested in MND in clinical practice but
are recommended by EFNS. Nonpharmacological treatment
modalities should first be deployed before pharmacological
interventions are introduced if symptoms do not improve. These drugs should be used with caution in MND patients
as they can cause depression of respiration and worsening Steinhansen et al. [99] evaluated a group of seriously ill
patients, recently bereaved patient relatives, physicians, and
other healthcare providers in a study to determine the factors
considered to be important at end-of-life. All respondents
agreed that naming a decision maker, maintaining one’s
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NNT/NL : 2022AIXM0094/012ED352 DES/IRESNE/DER/SPESI/LP2E 2022-0007-DO PhD THESIS Soutenue à Aix-Mars
eille Université le
30/03/2022
par Augusto Di Chicco Characterization of a stilbene organic scintillator for use as a broadband neutron spectrometer in mixed radiations fields Discipline Physique et Sciences de la Matière Composition du jury
Stefan OBERSTEDT Physicist at the European Commission JRC Spécialité Instrumentation Sara POZZI École doctorale Ecole doctorale 352 – Physique et Sciences de la Matière Xavier LEDOUX Laboratoire/Partenaires de recherche CEA / DES / IRESNE / DER / SPESI / LP2E (Laboratoire des programmes expérimentaux et d'essais en sûreté) IRSN / PSE-SANTE / SDOS / LMDN (Laboratoire de micro-irradiation, de métrologie et de dosimétrie des neutrons) Rapporteur Rapporteuse Professor at University of Michigan Examinateur Research Engineer at GANIL, CEA Paolo MUTTI Examinateur Head of Scientific Computing Division at ILL Mossadek TALBY Président du jury Professor at Aix-Marselle University Brian STOUT Directeur de thèse Professor at Aix-Marselle University Alix SARDET Encadrante/Invitée Research Engineer at CEA (Cadarache) Michael PETIT Encadrant/Invitée Researcher at IRSN (Cadarache) Robert JACQMIN Research Director at CEA (Cadarache) Encadrant/Invitée
Cette page est laissée blanche intentionnellement AFFIDAVIT I, undersigned, Augusto DI CHICCO, hereby declare that the work presented in this manuscript is my own work, carried out under the scientific direction of Alix SARDET, Michael PETIT, Robert JACQMIN and Brian STOUT, in accordance with the principles of honesty, integrity and responsibility inherent to the research mission. The research work and the writing of this manuscript have been carried out in compliance with both the French national charter for Research Integrity and the Aix-Marseille University charter on the against plagiarism. This work has not been submitted previously either in this country or in another country in the same or in a similar version to any other examination body. Place Aix-en-provence, date 30 mars 2022
Cette
œuvre est mise
à
disposition selon les termes de la Licenc
e Creative Commons Attribution Pas d'Utilisation Commerciale Pas de Modification 4.0
International. i Cette page est laissée blanche intentionnellement ii
“It
does not depend
, then, merely
upon what
one
se
es, but
what
one sees depends upon
how
one sees; all observation is not just
a
receiving, a discovering, but also
a
bring
ing
forth
” (Søren Kierkegaard) iii Cette page est laissée blanche intentionnellement iv
LIST OF PUBLICATIONS AND PARTECIPATION IN CONFERENCES
1) List of publications made within the framework of the thesis project: 1. “Investigation of the neutron-gamma ray discrimination performance at low neutron energy of a solution-grown stilbene scintillator”, Augusto DI CHICCO (CEA,AMU), Michael PETIT (IRSN), Robert JACQMIN (CEA), Vincent GRESSIER (IRSN), Brian STOUT (AMU). EPJ Web of Conferences 225, 04013 (2020) 2) List of submited publications: 1. “Gamma-response characterization of a solution-grown stilbene scintillators in the 59 keV 4.44.MeV energy range; an alternative low-resolution gamma spectrometer”, Augusto DI CHICCO (CEA,AMU), Alix SARDET (CEA), Michael PETIT (IRSN), Robert JACQMIN (CEA), Vincent GRESSIER (IRSN), Brian STOUT (AMU). Target Journal: Nuclear Instruments and Methods in Physics Research Section A. (2022) 2. “Extended time-of-flight measurements down to 100 keV at the AMANDE facility with a stilbene scintillator”, Michael PETIT (IRSN), Augusto DI CHICCO (CEA,AMU), Robert JACQMIN (CEA), Alix SARDET (CEA), Brian STOUT (AMU), Richard Babut (IRSN). For NEUDOS14 (Neutron and Ion Dosimetry Syposium), Krakow (Poland) April 2022 3) Participation in conferences and summer schools during the thesis period: a. ANIMMA (Advancements in Nuclear Instrumentation Measurement Methods and their Applications) 2019, Portorož (Slovenia); b. SANDA (SUPPLYING ACCURATE NUCLEAR DATA FOR ENERGY AND NON-ENERGY APPLICATIONS) General meeting on Zoom le 9/02/2021: “Progress on the commissioning of a compact broad-band fast neutron spectrometer”. v ABSTRACT
Many nuclear applications require a precise characterization of the produced neutron field. However, this neutron production is accompanied by a photonic field. The detectors used need to cover a large energy range (several tens of MeV) with a good detection efficiency, an energy resolution lower than 10%, but also to have a relatively stable response to variations in the measurement environment, while being insensitive to gamma radiation or able to discriminate it from neutrons. Crystalline organic scintillators can meet most of the above criteria but, until recently, they could not be produced in large volumes. The development of crystal growth techniques has allowed the production of a 25.4 × 25.4 mm3 stilbene single crystal, which is able, as shown in previous work, to measure neutrons down to 565 keV and is suitable for the neutron characterization of irradiation locations in research reactors. This thesis work follows up on this work, with the primary objective of establishing the neutron response matrix of the detector to determine its suitability as a broadband neutron spectrometer in mixed radiation fields. Firstly, the photon response of the detector between 0.059 MeV and 4.5 MeV has been determined using two methods. The first one is based on a series of direct irradiations with gamma sources (classical procedure) and the second one with the same setup but adding a coincidence measurement to select only part of the response (technique requiring long measurements). The two techniques gave consistent calibration results. With the energy resolution parameters defined, the photon response matrix was calculated by simulation and tested on gamma ray sources using the GRAVEL and MAXED codes. Despite a difficulty to compute the absolute intensities, the detector is able to identify the main gamma peaks of the spectrum and can be used as a lowresolution gamma-ray spectrometer. The neutron characterization of the detector in the energy range 0.5 MeV and 17 MeV was carried out with three different neutron field production facilities. The first two, the AMANDE facility (IRSN/France) and the PTB (Germany) Tandetron, produce monoenergetic neutron beams. The measurements carried out at AMANDE allowed to study the main characteristics of the neutron response of the stilbene and the monoenergetic neutron fields of the PTB were used for comparative purposes. These measurements allowed us to study the discrimination capabilities between neutrons and gamma rays, to determine the exploitable energy range and to build stochastic models using the MCNPX-PoliMi and Geant4 codes. The PTB cyclotron produces a polyenergetic neutron field, up to 16.5 MeV for this study, thus covering the energy range inaccessible with monoenergetic beams and allowing the constitution of the complete experimental response matrix of the detector in a single measurement. The experimental response matrix allowed unfolding mono and polyenergetic spectra with results consistent with the expected spectra. However, the normalization in absolute could not be calculated due to a lack of time. In addition, different attempts were made to simulate the anisotropy of the stilbene. The first results obtained are encouraging. To complete this work, the experimental response matrix should be finalized, as well as the understanding and modeling of the anisotropy phenomenon. Nevertheless, it is possible to conclude that stilbene can be used as a broadband neutron spectrometer down to 0.7 MeV.
Keywords: organic scintillator, stilbene, neutron spectrometry, PSD, time of flight, MCNP, Geant4, response matrix, unfolding vi
RESUME
De nombreuses applications nucléaires nécessitent une caractérisation précise du champ neutronique produit. Cependant, cette production de neutrons s’accompagne d’un champ photonique. Les détecteurs utilisés doivent couvrir une large gamme en énergie (plusieurs dizaines de MeV) avec une bonne efficacité de détection, une résolution en énergie inférieure à 10%, mais aussi avoir une réponse relativement stable vis-à-vis des variations de l’environnement de mesure, tout en étant insensible au rayonnement gamma ou capable de le discriminer des neutrons. Les scintillateurs organiques cristallins permettent de répondre à l’essentiel des critères cités mais, jusqu’à récemment, ils ne pouvaient être produits en grands volumes. Le développement de techniques de croissance des cristaux a permis la production d’un monocristal de stilbène de 25,4 × 25,4 mm3 dont des travaux antérieurs ont prouvé qu’il permet de mesurer des neutrons jusqu’à 565 keV, au moins, et qu’il est adapté à la caractérisation neutronique d’emplacements d’irradiation dans des réacteurs de recherche. Ce travail de thèse fait suite à ces travaux, avec comme objectif premier l’établissement de la matrice de réponse neutrons du détecteur afin de déterminer son adéquation comme spectromètre neutronique à large bande dans des champs de rayonnement mixtes. Dans un premier temps, la réponse aux photons du détecteur entre 0,059 MeV et 4,5 MeV a été établie au moyen de deux méthodes. La première est basée sur une série d’irradiation directes avec des sources gamma (procédure classique) et la seconde avec le même dispositif mais en sélectionnant une partie de la réponse en imposant une coïncidence (technique nécessitant des mesures longues). Les deux techniques ont donné des résultats d’étalonnage concordants. Les paramètres de résolution en énergie définis, la matrice de réponse aux photons a été calculée par simulation puis testée sur des sources gamma en utilisant les codes GRAVEL et MAXED. Malgré une difficulté pour calculer les intensités en absolue, le détecteur est capable d’identifier les principales raies gamma du spectre et peut être utilisé comme un spectromètre gamma à faible résolution. La caractérisation neutronique du détecteur dans la gamme d’énergie 0.5 MeV et 17 MeV a été réalisée avec trois installations de production de champs neutroniques différentes. Les deux premières, l’installation AMANDE (IRSN/France) et de le Tandetron de la PTB (Allemagne), produisent des faisceaux de neutrons monoénergétiques. Les mesures effectuées sur AMANDE ont permis d'étudier les principales caractéristiques de la réponse neutronique du stilbène et les champs de neutrons monoénergétique de la PTB ont été utilisés dans un but comparatif. Ces mesures nous ont permis d'étudier les capacités de discrimination entre les neutrons et les rayons gamma, la gamme d'énergie exploitable et de construire des modèles stochastiques en utilisant les codes MCNPX-PoliMi et Geant4. Le cyclotron de la PTB permet de produire un champ neutronique polyénergétique (jusqu’à 16,5 MeV pour cette étude), couvrant ainsi la gamme en énergie inaccessible au moyens de faisceaux monoénergétiques et permettant la constitution de la matrice de réponse expérimentale complète du détecteur en une unique mesure. La matrice de réponse expérimentale a permis de déconvoluer les spectres mono et polyénergétiques avec des résultats cohérent avec les spectres attendus. Cependant la normalisation en absolue n’a pu être calculée par manque de temps. Par ailleurs, différentes tentatives ont été effectuées afin de simuler l’anisotropie du stilbène. Les premiers résultats obtenus sont encourageants. Pour finaliser ce travail, la matrice de réponse expérimentale devra être finalisée, de même que la compréhension et la modélisation du phénomène d’anisotropie. Néanmoins, il est possible de conclure que le stilbène peut être utilisé comme spectromètre neutronique à large bande jusqu'à 0.7 MeV.
Mots clés : scintillateur
organi
que, stilbène, spectrométr neutr
onique
, PSD, temps de vol, MCNP, Geant4, matrice de réponse, déconvolution vii
RIASSUNTO
Molte applicazioni nucleari necessitano di una caratterizzazione precisa del campo neutronico prodotto. Tuttavia, questa produzione di neutroni è accompagnata da un campo di fotoni. I detector utilizzati devono coprire un'ampia gamma di energia (diverse decine di MeV) con una buona efficienza di rilevamento, una risoluzione energetica inferiore al 10%, ma devono anche avere una risposta relativamente stabile alle variazioni dell'ambiente di misura, pur essendo insensibili alla radiazione gamma o capaci di discriminarla dai neutroni. Gli scintillatori organici cristallini soddisfano la maggior parte di questi criteri ma, fino a poco tempo fa, non potevano essere prodotti in grandi volumi. Lo sviluppo delle tecniche di crescita del cristallo ha permesso la produzione di un cristallo singolo di stilbene di 25,4 × 25,4 mm3, che ha dimostrato in lavori precedenti di essere in grado di misurare neutroni fino a 565 keV ed adatto alla caratterizzazione neutronica nei luoghi di irradiazione dei reattori di ricerca. Questo lavoro di tesi segue questo lavoro, con l'obiettivo primario di stabilire la matrice di risposta neutronica del rivelatore per determinare la sua idoneità come spettrometro neutronico a banda larga in campi di radiazione mista. In una prima fase, la risposta fotonica del rivelatore tra 0,059 MeV e 4,5 MeV è stata stabilita per mezzo di due metodi. La prima si basa su una serie di irradiazioni dirette con fonti gamma (procedura classica) e la seconda con lo stesso dispositivo ma selezionando una parte della risposta imponendo una coincidenza (tecnica che richiede lunghe misurazioni). Entrambe le tecniche hanno dato risultati di calibrazione coerenti. Con i parametri di risoluzione energetica definiti, la matrice di risposta fotonica è stata calcolata per simulazione e poi testata su sorgenti di raggi gamma usando i codici GRAVEL e MAXED. Nonostante la difficoltà nel calcolare le intensità assolute, il rivelatore è in grado di identificare i principali picchi dei raggi gamma nello spettro e può essere utilizzato come uno spettrometro gamma a bassa risoluzione. La caratterizzazione neutronica del detector nell'intervallo di energia compreso tra 0,5 MeV e 17 MeV è stata effettuata con tre diversi impianti di produzione di campi neutronici. I primi due, l'impianto AMANDE (IRSN/Francia) e il Tandetron della PTB (Germania), producono fasci di neutroni monoenergetici. Le misure effettuate ad AMANDE ci hanno permesso di studiare le principali caratteristiche della risposta neutronica dello stilbene e i campi neutronici monoenergetici del PTB sono stati utilizzati a scopo comparativo. Queste misure ci hanno permesso di studiare le capacità di discriminazione tra neutroni e raggi gamma, l'intervallo di energia sfruttabile e di costruire modelli stocastici utilizzando i codici MCNPX-PoliMi e Geant4. Il ciclotrone della PTB permette la produzione di un campo neutronico polienergetico (fino a 16,5 MeV per questo studio), coprendo così la gamma di energia inaccessibile per mezzo di fasci monoenergetici e permettendo la costituzione della completa matrice di risposta sperimentale del rivelatore in una sola misura. La matrice di risposta sperimentale ha permesso la deconvoluzione degli spettri mono e polienergetici con risultati coerenti con gli spettri previsti. Tuttavia, per mancanza di tempo, non è stato possibile calcolare la normalizzazione in termini assoluti. Inoltre, sono stati fatti vari tentativi per simulare l'anisotropia dello stilbene. I primi risultati ottenuti sono incoraggianti. Per finalizzare questo lavoro, la matrice di risposta sperimentale dovrà essere completata, così come la comprensione e la modellizzazione del fenomeno dell'anisotropia. Tuttavia, si può concludere che lo stilbene può essere utilizzato come spettrometro di neutroni a banda larga fino a 0,7 MeV. Parole chiave: scintillatore organico, stilbene, spettrometria neutronica, PSD, tempo di volo, MCNP, Geant4, matrice di risposta, deconvoluzione
viii RESUME ETENDU 1. INTRODUCTION
La spectroscopie neutronique rapide présente un intérêt particulier dans de nombreux domaines scientifiques, parmi lesquels la détection de matériaux dangereux pour la sécurité nationale, l'imagerie neutronique, le contrôle des neutrons produits par la fusion, les applications médicales telles que la tomographie neutronique, et la caractérisation des neutrons produits par le rayonnement cosmique pour la protection des astronautes dans l'espace. Ces nombreuses applications nucléaires nécessitent une caractérisation précise du champ neutronique produit. Cependant, cette production de neutrons s’accompagne d’un champ photonique. Le spectromètre neutronique idéal doit avoir les propriétés suivantes : 1. Petite taille (< Ø60 mm) afin de pouvoir être utilisé dans des environnements où l'espace est restreint (par exemple à l'intérieur d'un réacteur nucléaire). 2. Couvrir une gamme d'énergie aussi large que possible (entre 0,1 jusqu’à 20 MeV). 3. Insensibilité aux rayons gamma ou capacité de séparer/discriminer les contributions produites par ces particules dans la gamme d'énergie exploitable. 4. Bonne résolution en énergie (< 10%) sur toute la gamme afin de distinguer les différents groupes d'énergie des photons et des neutrons. 5. Haute efficacité de détection afin de limiter le temps de mesure dans les environnements à faible fluence neutronique. 6. Stable et peu affectée par les variations de fluence neutronique, de température, etc., pendant les opérations d'acquisition de données. Ce travail de thèse vise à la caractérisation neutronique et photonique d'un scintillateur organique basé sur un cristal Ø25.4 x 25.4 mm3 de stilbène, afin de démontrer son utilisation comme spectromètre à large bande dans des champs de rayonnement mixtes. Pour atteindre cet objectif, il est nécessaire d'établir les matrices de réponse du stilbène afin qu'elles puissent être utilisées dans le processus de déconvolution pour essayer de calculer les spectres d'énergie qui ont généré la réponse du stilbène.
2. LES SCINTILLATEURS ORGANIQUES APPLIQUES A LA SPECTROMETRIE NEUTRONIQUE RAPIDE
Les scintillateurs organiques sont constitués de composés aromatiques (anneaux de carbone avec de l'hydrogène), capables de produire de la lumière dans le spectre visible lorsqu'ils interagissent avec un rayonnement ionisant. Les particules chargées de recul, produites par les neutrons et les rayons gamma, interagissent avec les électrons des molécules environnantes, les amenant à un état excité. Pour revenir à leur état fondamental, les molécules vont émettre des photons dans le spectre visible, d'où le nom de "scintillateur". Afin d'extraire un signal lisible et analysable, la lumière produite doit être convertie en un signal électrique. Cette opération est réalisée à l'aide d'un photomultiplicateur (PMT), directement attaché au scintillateur. Une photocathode convertit la lumière en photoélectrons, qui sont ensuite amplifiés par plusieurs étages de dynodes avant de sortir du circuit sous forme de signal électrique à la sortie de l'anode du PMT. Ce signal électrique sera lu et analysé par un système d'acquisition afin d'extrapoler les informations sur la particule qui a généré le signal. La lumière produite dans les scintillateurs organiques dépend de l'énergie déposée par les particules chargées et de leur pouvoir d'ionisation. Dans le cas des électrons (produit par les rayons gamma), ils présentent une production de lumière linéaire pour des énergies supérieures à 0,125 MeV. Pour cette raison, l'unité de mesure pour quantifier la lumière est le MeV équivalent électron (MeVee), 1 MeVee correspond à la lumière produite par le dépôt d'énergie d'un électron de 1 MeV. Dans le cas d'ions plus lourds (protons, particules alpha, etc.) ix produits par des interactions neutroniques, la lumière produite est non linéaire en fonction de l'énergie déposée par la particule. Pour cette raison, afin de paramétrer correctement la réponse des particules de recul produites par les neutrons, il est nécessaire d'utiliser des formules semi-empiriques qui prennent en compte les effets de la non-linéarité. La Figure 1 montre la paramétrisation de la quantité de lumière pour les électrons (réponse linéaire) et les protons (réponse non linéaire) d'un scintillateur liquide organique.
Figure 1 : Quantité de lumière produite par un scintillateur liquide en fonction de l'énergie déposée par la particule incidente : électrons (triangles noirs pleins) et protons (cercles noirs pleins). La forme du signal produit par les rayons gamma et les neutrons est différente en raison des différents mécanismes d'excitation et de désexcitation de molécules organiques. Par conséquent, il est possible d'analyser cette forme pour identifier le type de particule de recul et donc la particule incidente. L'une des méthodes les plus utilisées pour mettre en œuvre la discrimination de la forme des impulsions, appelé en anglais Pulse Shape Discrimination (PSD) est la méthode de comparaison des charges (charge comarison method : CCM). Une autre façon d'aider à discriminer les neutrons des rayons gamma est l'utilisation de la technique du temps de vol (TOF). En effet, l'identification des particules incidentes est rendue possible par la différence de vitesse entre les deux types de particules. Les rayons gamma, étant une radiation électromagnétique, se déplacent dans l'air entre le point source et le détecteur à la vitesse de la lumière, alors que la vitesse des neutrons dépend de leur énergie cinétique. Cependant, au moins deux détecteurs sont nécessaires pour utiliser cette technique. L'opérateur mathématique qui relie le flux incident à la réponse du détecteur générée par le flux est appelé matrice de réponse. La matrice de réponse pour une particule donnée contient toutes les réponses normalisées du détecteur dans la gamme d'énergie exploitable. Traditionnellement, la matrice de réponse est construite à l'aide de simulations de Monte Carlo, mais grâce à la technique du temps de vol, il est également possible de la mesurer (uniquement dans le cas des neutrons). La Figure 2 montre une représentation graphique de la matrice de réponse neutronique d'un scintillateur organique liquide EJ-301. La matrice de réponse est fondamentale pour l'opération de déconvolution des réponses expérimentales d'un détecteur, c'est-à-dire l'opération qui permet de calculer le spectre 'énergie incidente qui a généré la réponse dans le dispositif. x Figure 2 : Représentation graphique de la matrice de réponse aux neutrons d'un scintillateur organique liquide EJ-301.
Parmi les différents types de scintillateurs organiques, les scintillateurs cristallins, malgré des coûts de production plus élevés et les effets de l'anisotropie de la réponse neutronique, présentent divers avantages tels qu'une plus grande quantité de lumière produite, excellente capacité de discrimination des neutrons et des rayons gamma, une plus grande résistance aux fluences neutroniques élevées et aux températures élevées (entre 77 et 107 °C). Grâce à la nouvelle technique de croissance en solution, il a été possible de réduire les coûts de production des scintillateurs organiques basés sur des molécules de stilbène. Pour ces raisons, L. Dioni a choisi un cristal de stilbène cylindrique de Ø25.4 x 25.4 mm3 comme candidat pour être utilisé comme spectromètre à neutrons rapides pour le réacteur expérimental MASURCA. L. Dioni a eu l'occasion de prouver que le stilbène capable de mesurer des neutrons jusqu’à 565 keV et qu’il est adapté à la caractérisation neutronique d’emplacements d’irradiation dans des réacteurs de recherche. Ce travail de thèse fait suite aux travaux de L. Dioni, avec comme objectif la caractérisation de la réponse photonique et principalement de la réponse neutronique pour établir la matrice de réponse aux neutrons du détecteur afin de déterminer son adéquation comme spectromètre neutronique à large bande dans des champs de rayonnement mixtes. Dans ce travail de thèse, les données expérimentales ont été acquises à l'aide du système d'acquisition numérique CAEN DT5730, tandis que le programme ROOT développé par le CERN de Genève a été choisi pour l'analyse des données. 3. CARACTERISATION PHOTONIQUE DU STILBENE
Pour la caractérisation photonique du stilbène, deux méthodes d'étalonnage de l'énergie ont été réalisées à l'aide de sources gamma couvrant une gamme d'énergie comprise entre 0,059 et 4,5 MeV. La première procédure est basée sur une série d'irradiations directes du stilbène avec des sources gamma. Grâce à ces mesures, il a été possible d'établir la résolution énergétique du dispositif afin de les intégrer dans les simulations MCNPX. La procédure d'étalonnage a été vérifiée en comparant les spectres expérimentaux à ceux obtenus à partir des simulations. Les résultats ont montré que la réponse aux rayons gamma du stilbène est linéaire uniquement dans la plage d'énergie comprise entre 0,3 et 2 MeV. Alors que pour les énergies inférieures à 0,3.MeV et supérieures à 2 MeV, la réponse n'est plus linéaire.
xi La
seconde
méthode d'étalonnage est basée sur la méthode des coïncidences, en utilisant un second cristal de Ø50.8 x 50.8 mm3 de stilbène pour sélectionner les rayons gamma qui sont diffusés dans la trajectoire qui coupe les deux détecteurs, c'est-à-dire à 90° pour la configuration expérimentale mise en œuvre. Cette technique nécessite beaucoup plus de temps que les mesures directes pour obtenir de bonnes statistiques dans les spectres expérimentaux, mais permet un étalonnage plus
rapide
et
dépend moins
de la comparaison
avec les spectres obtenus par simulation.
La comparaison entre les deux méthodes d'étalonnage a montré que les mesures directes permettent un étalonnage en énergie fiable et rapide et peuvent être utilisées en routine sans introduire de biais significatif. Grâce à la validation de la résolution énergétique du stilbène, il a été possible construire la matrice de réponse aux rayons gamma du stilbène dans la gamme d'énergie comprise entre 0,01 et 7,3 MeV, voir la Figure 3. Figure 3 : Matrice de réponse aux rayons gamma du stilbène entre 0,01 et 7,3 MeV d'énergie avec 1024 bins entre 0 et 8 MeVee. La matrice de réponse photonique a été testée avec les deux codes de déconvolution choisis pour ce travail, l'algorithme itératif GRAVEL (basé sur la méthode des moindres carrés non linéaires) et MAXED (basé sur la théorie bayésienne couplée à la méthode de l'entropie). Les deux codes ont donné des résultats cohérents avec les pics théoriques des sources testées, étant capables de reconnaître les pics principaux d'une source complexe comme celle du 152Eu (voir la Figure 4). Figure 4 : Spectres déconvolué du 152Eu avec GRAVEL (ligne pleine bleue) et MAXED (ligne pleine rouge), comparés aux pics gamma théoriques (ligne pointillée noire). xii 4. CARACTERISATION NEUTRONIQUE DU STILBENE
La caractérisation de la réponse neutronique du stilbène a été réalisée à l'aide de trois installations différentes de production de champs neutroniques. Les deux premières, l’installation AMANDE (IRSN/France) et de le Tandetron de la PTB (Allemagne), produisent des faisceaux de neutrons monoénergétiques. Les mesures effectuées sur AMANDE ont permis d'étudier la réponse neutronique du stilbène dans la gamme d'énergie comprise entre 0,481 et 17 MeV, avec l'absence des énergies comprises entre 5 et 13,713 MeV, en utilisant également pour certaines énergies la technique du temps de vol. Ces mesures ont permis d'étudier la gamme d'énergie exploitable et les capacités discriminantes du stilbène, qui est capable de séparer correctement et sans difficulté les contributions des neutrons et des rayons gamma. En outre, ces mesures ont permis la paramétrisation de la réponse neutronique (pour les protons de recul et les particules alpha) afin de pouvoir construire la réponse neutronique du stilbène à l'aide des codes Monte Carlo MCNPX-Polimi (avant) et Geant4 (pour une plus grande précision des données). La comparaison des spectres expérimentaux avec ceux obtenus à partir des simulations a montré que les simulations présentent deux types d’écarts. La première concerne le fait que la forme du spectre simulé ne présente pas la même non-linéarité que le spectre expérimental, cet effet pourrait être lié à l'anisotropie de la réponse neutronique du stilbène. La seconde concerne les contributions des particules alpha produites par les neutrons à haute énergie, qui pourraient être liées à des données incorrectes des sections efficaces utilisées pour simuler la réponse du stilbène aux neutrons (JENDL.4.0u). Les champs neutroniques monoénergétiques du Tandet de la PTB ont été utilisés uniquement dans un but comparatif. Le troisième champ neutronique est un spectre polyénergétique (énergie maximale de 16,5 MeV) produit par le cyclotron de la PTB, qui est capable de couvrir une gamme d'énergie inaccessible aux réactions classiques de production de neutrons monoénergétiques, il est communément appelé le spectre blanc. En utilisant la technique du temps de vol, il a été possible d'obtenir une paramétrisation précise et complète de la réponse donnée par les protons de recul et les particules alpha. Cependant, comme le montre la Figure 5, il a été constaté que la réponse des protons de recul présente une tendance non linéaire à la fois aux basses énergies (entre 0 et 2,5 MeV) et aux hautes énergies (entre 10 et 16,5 MeV).
Figure 5 : Comparaison entre la quantité de lumière produite par le stilbène à la PTB (carrés noirs vides) et à AMANDE (carrés noirs pleins).
xiii En utilisant des mesures effectuées avec le spectre blanc de la PTB en changeant l'orientation du cristal, on a essayé d'introduire un terme d'anisotropie dans la paramétrisation de la sortie de lumière. Les simulations Geant4 réalisées avec ce terme correctif sont en meilleur accord avec les données expérimentales, même si des déviations flagrantes persistent. Ces déviations sont probablement causées par une paramétrisation incomplète de l'anisotropie (échantillonnage angulaire limité). Néanmoins, ce résultat prouve que l'anisotropie pourrait être modélisée avec succès, à condition de disposer d'un ensemble de données ultérieures de mesures angulaires. Cependant, l'objectif principal des mesures avec le spectre blanc de la PTB était de mesurer la matrice de réponse neutronique du stilbène. La matrice de réponse mesurée couvre une gamme d'énergie comprise entre 0,69 (énergie minimale détectable du stilbène) et 16,51 MeV, voir la Figure 6. Une deuxième matrice de réponse a été construite à l'aide de Geant4, qui couvre une gamme d'énergie comprise entre 0,01 et 17,02 MeV. Cependant, la matrice de réponse simulée, contrairement à la matrice expérimentale, ne tient pas compte des effets réels de l'anisotropie des neutrons et des contributions apportées par la production de particules alpha de recul.
Figure 6 : Matrice de réponse neutronique expérimentale du Stilbène dans la gamme d'énergie comprise entre 0,67 et 16,51 MeV avec 1204 bins entre 0 et 9,12 MeVee.
Les matrices de réponse expérimentales et simulées ont été utilisées pour déconvoluer les spectres neutroniques monoénergétiques et polyénergétiques (AmBe, spectre neutronique blanc de la PTB). Pour les premiers, les spectres sont relativement simples et le code de déconvolution GRAVEL a permis de déterminer avec une bonne précision le spectre incident. Pour les spectres polyénergétiques plus complexes, les résultats ont montré que les deux matrices de réponse sont capables de calculer la forme des spectres incidents. Dans le cas de la source AmBe, les pics principaux de la source ont été correctement identifiés malgré des intensités relatives différentes par rapport à la ISO 8529, voir la Figure 7. L'origine de ces écartes peut se trouver dans l'imprécision du code de déconvolution GRAVEL. Pour cette raison, plus d'efforts devraient être concentrés sur le code MAXED (ou d'autres) qui semble plus optimal pour la déconvolution des spectres. xiv Figure 7 : Comparaison entre les spectres déconvolués de la source AmBe en utilisant la matrice de réponse (RM) de Geant4 (ligne plaine rouge) et la matrice de réponse expérimentale (ligne plaine noire) avec le spectre de référence ISO-8529 (ligne plaine bleue).
5. CONCLUSION GENERALE
L'objectif principal de cette thèse était de caractériser la réponse d'un scintillateur organique basé sur un cristal de stilbène cylindrique Ø25.4 x 25.4 mm3, afin d'évaluer ses performances en tant que spectromètre neutronique à large bande dans des champs de rayonnement mixtes. À cette fin, les réponses photoniques et neutroniques du détecteur ont été établies et utilisées comme paramètres d'entrée, en essayant de déployer des spectres bien connus. Globalement, l'objectif principal a été atteint. Cependant, les divergences entre les fluences calculées et estimées pour les rayons gamma et les neutrons doivent être résolues afin d'utiliser le stilbène comme spectromètre. En outre, des mesures supplémentaires sont nécessaires pour compléter les études de l'anisotropie neutronique du stilbène afin de les intégrer correctement dans les simulations de Geant4. xv ACKNOWLEDGEMENTS
“Gratus animus virtus est non solum maxima sed etiam mater virtutum omnium” (Marco Tullio Cicerone)
xvi INDEX Affi
davi
t...................................................................................................................................................................i List of publications and partecipation in conferences........................................................................................... v Abstract................................................................................................................................................................. vi Résumé................................................................................................................................................................. vii Riassunto............................................................................................................................................................. viii Résumé étendu...................................................................................................................................................... ix Acknowledgements............................................................................................................................................. xvi List of figures....................................................................................................................................................... xxi List of Tables.....................................................................................................................................................xxviii Glossary.............................................................................................................................................................. xxx
1 2 Introduction....................
................
................................................
................................................................1
1.1 Context of the thesis project..................................................................................................................2 1.2 Fast neutron spectrometry needs..........................................................................
................................
3 1.3 Thesis objectives.....................
................
................................
................................................................
5 1.4 Manuscript content
................
................
................................................................................................
6
Organic
scint
illators
applied
to
fast neutro
n spectr
ometry............
................................
................
................
9 2.1 Neutral particles interactions with matter.......................................................................................... 10 Gamma Rays.................................................................................................................................... 10 Neutrons.......................................................................................................................................... 13 2.2 Organic scintillators............................................................................................................................. 16 Scintillation theory...........................................................................................................
................
17 Quantification of the produced light...............................................................................
................
19 Discrimination between neutrons and gamma rays....................................................................... 21 2.2.3.1 Charge Comparison Method................................................................................................... 21 2.2.3.2 Improving neutron-gamma discrimination using the time of flight technique....................... 23 The detector’s response matrix....................................................................................................... 24 Theory of the anisotropy in crystalline organic scintillators........................................................... 26 2.3 Motivations for choosing a Ø25.4 x 25.4 mm3 stilbene crystal................................
........................... 28 xvii Scientific background on stilbene crystals....................................................................................... 28 Technological choices...................................................................................................................... 30 First performance assessment......................................................................................................... 31 2.4 3 Chapter conclusions............................................................................................................................
37 Data analysis framework and Experimental facilities..................
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................................
................
40
3.1 Data acquisition
...................................................................................................................................
41 The CAEN DT5730 digital acquisition system.................................................................................. 41 The DPP-PSD firmware and the CoMPASS program
........................................................................ 42 3.1.2.1 Input settings........................................................................................................................... 43 3.1.2.2 Discriminator
settings
.............................................................................................................. 44
3.1.
2.3
QDC settings
............................................................................................................................ 44 3.1.2.4
Time
Selection and On
board
coincidence............................................................................... 45 3.1.2.5 Acquisition settings.................................................................................................................. 45 3.2 Data analysis using the ROOT framework........................................................................................... 46 3.3 Experimental facilities.........................................................................................................................
48
AM
ANDE Facility at IRSN.................
................................................................................
................ 48
PIAF
at
the
PTB................................................................................................................................ 50 3.4 4 3.3.2.1 Monoenergetic neutron.......................................................................................................... 50 3.3.2.2 White neutron spectrum.........................................................................................................
51 Chapter conclusions............................................................................................................................ 53 Gamma response of the stilbene detector.................................................................................................. 55
4.1 Experimental and simulation tools...................................................................................................... 56 Gamma sources............................................................................................................................... 56 Direct measurements...................................................................................................................... 57 4.1.2.1
Experimental
setup.................................................................................................................. 57 4.1.2.2 MCNP model............................................................................................................................ 58 Coincidence measurements............................................................................................................ 59 4.1.3.1 Experimental setup.................................................................................................................. 59 4.1.3.2 MCNP model............................................................................................................................ 61 4.2 Energy resolution calculation.............................................................................................................. 62 4.3 Energy calibration validation............................................................................................................... 64 Direct measurements...................................................................................................................... 64 Co
incidence
measurements............
................................................................
................................ 69 Performance comparison of the calibration methods.................................................................... 74
xviii
5 4.4
Gamma ray intrinsic
efficiency............................................................................................................ 75 4.5 Construction of the gamma rays response matrix with MCNPX......................................................... 76
4.6 Chapter conclusions............................................................................................................................ Direct gamma ray MCNPX input............................................................................................................ 166 B. Coincidence gamma ray MCNPX-Polimi input....................................................................................... 167 C. Geant4 inputs for the stilbene neutron simulation............................................................................... 169 D. Geant4 general particle source input for 5 MeV AMANDE simulation................................................. 192 E. Code for the construction of the neutron PID simulated with Geant4.................................................
195 Appendix B
......................................................................................................................................................... 200 A. Results on anisotropy of recoil alpha particles obtained at PTB........................................................... 200 B. Neutron response in the energy range between 0.1 to 1.2 MeV......................................................... 204 xx
LIST OF
FIGURES
Figure 1: Energy domain of interest
for the SPECTRAL
project
and associated neutron detectors......................4 Figure 2: Ideal energy distribution spectrum produced by the photoelectric effect induced by a gamma ray having an incident energy Eγ.............................................................................................................................. 10 Figure 3: Graphic representation of Compton scattering where an incident photon (red wave) after a collision changes direction θ (green wave) and deposits part of its energy to a recoil electron Ee (yellow circle)......... 10 Figure 4: Ideal energy distribution spectrum produced by the Compton scattering induced by a gamma ray having an incident energy Eγ.............................................................................................................................. 11 Figure 5: Ideal energy distribution spectrum produced by the pair production reaction induced by a gamma ray having an incident energy Eγ. In this graph, the two 511 keV gammas resulting from positron annihilation are not detected........................................................................................................................................................ 11 Figure 6: Predominance of the three types of gamma ray interactions as a function of their incident energy and the atomic number of the target material. In the case of organic scintillators Compton effect is dominant as they are compounds of hydrogen Z=1 and carbon Z=6...................................................................................... 12 Figure 7: (Left) Ideal energy distribution spectrum produced by a gamma ray having an incident energy Eγ ≫ 2m0c2. (Right) ideal energy distribution obtained from a detector operating with (solid black line) and without (dashed black line) the coincidence mode [3]..................................................................................................... 12 Figure 8: Hydrogen and carbon neutron cross section from the nuclear library JEFF-3.3 [8]. The cross section of 12 C(n,n’)3α comes from the CENDL-3.2 library [9] as it has not been evaluated in JEFF-3.3............................ 13 Figure 9: Neutron elastic scattering in the laboratory system............................................................................ 14 Figure 10: Isotropic ideal energy distribution probability P(Er) of a proton recoil particle generated by an incident neutro
n En............................................................................................................................................
14 Figure 11: Proton energy distribution probabilities influenced by bias factors such as: scattering on carbon atoms (a), nonlinearity (b) and energy resolution (c)......................................................................................... 15 Figure 12: Schematic representation of the experimental configuration of a scintillator plus its PMT, connected to the high-voltage power supply and acquisition system.................................................................................. 16 Figure 13: Perrin-Jablonski diagram showing the various processes involved in the light production.............. 18 Figure 14: Graphical representation of the signal produced by a PMT attached to an organic scintillator with its components: excitation (full red line), prompt fluorescence (solid blue line), delayed fluorescence (dashed blue line), phosphorescence (full green line) and their sum (full black line).............................................................. 18 Figure 15: Shape of the signal generated by a stilbene crystal for different incident particles [23].................. 19 Figure 16: Light-output produced by a liquid LAB scintillator in function of the energy deposited by the incident particle: electrons (full black triangles) and protons (full black circles). Proton data have been fitted using equation (9) [32].................................................................................................................................................. 20 Figure 17: Example of a digitized pulse generated by the stilbene crystal with the two integrals: long gate Q L (red full line) and short gate QS (blue full line).................................................................................................... 21 Figure 18: Schematic representation of a two-dimensional histogram PSD vs QL (a). Results obtained with a deuterated stilbene subjected to a source of 252Cf. [33] (b)............................................................................... 22 Figure 19: Scheme of the PSD spectrum with the various variables used in the FOM calculation..................... 22 Figure 20: Comparison of calculated FOM for different types of organic scintillators [45]................................ 23 Figure 21: TOF spectrum generated by a 252Cf source with (red full line) and without (blue full line) a PSD selection to extract the neutron contribution [54]............................................................................................. 24 Figure 22: Graphical representation of the neutron response matrix of an EJ-301 liquid organic scintillator [61, p. 309].................................................................................................................................................................. 25 Figure 23: Graphical representation of crystal structure arrangements for various organic molecules according to the a (full red line), b (full green line) and c (full blue line) crystal axes [77]..................................................
26 xxi
Figure 24: Example of light-outputs produced by a 1 cm3 cubic trans
-stilbene crystal according its axes: a (red full line), b (blue full line) and c’ (green full line) [84].........................................................................................
27 Figure 25: Microscopic picture showing two stilbene crystals obtained with the solution-grown (left) and the melt-grown technique [96] (The arrow points to the tip of a vicinal sector typical for single crystals grown from solution by the dislocation mechanism).............................................................................................................. 28 Figure 26: Comparison between anthracene’s and stilbene’s response for electrons (a) and proton recoils (b) [100]..................................................................................................................................................................... 28 Figure 27: Stilbene spectra produced by a 60Co source (a) and its energy resolution (b) after been irradiated to different neutron fluences [104]......................................................................................................................... 29 Figure 28: Variation of the relative light produced by a stilbene detector in function of the temperature when it is irradiated with a 137Cs source [35]................................................................................................................... 29 Figure 29: Comparison of the light-output produced by a 137Cs source of different organic scintillators in function of their length (a) and comparison of the FOM of two stilbene crystals grown by different methods (b) [96]. 30 Figure 30: 25.4 mm × 25.4 mm3 cylindrical solution-grown stilbene detector with its PMT.............................. 31 Figure 31: Two-dimension PSD vs PID histogram of the stilbene (a) and the BC501A (b) subjected to 5 MeV neutrons [2]......................................................................................................................................................... 32 Figure 32: Comparison between calibrated PIDs subjected to 5 MeV neutrons in function of the crystal orientation: 0° (blue full line), -90° (orange full line) and 90° (yellow full line) [2]............................................. 32 Figure 33: Two-dimension histogram PSD vs PID for 565 keV neutrons (a) and its corresponding uncalibrated neutron PID after gamma contribution subtraction (b) [2]................................................................................. 33 Figure 34: Proton recoil light-output of a Ø1 x 1 cm3 melt-grown stilbene [2]................................................... 33 Figure 35: Comparison of unfolded PIDs measured at the LVR-15 reactor center channel for -930 V (blue solid line), -1000 V (red solid line) and -1100 V (yellow solid line) PMT’s volatages with reference simulation (dashed black line) [2]....................................................................................................................................................... 34 Figure 36: Section of the LR-0 nuclear reactor with B0, B1, and B2 measurement positions [2]...................... 35 Figure 37: Comparison between simulated (SIM) and experimental unfolded stilbene neutron spectra for measurement positions at B0 (a), B1 (b), and B2 (c) of the LR-0 reactor [2]...................................................... 36
Figure 38: DT5730 front face (154×50×164 mm3), the 8 MCX input channels are marked with Ch-i, on the right there is the B-type USB port PC connection........................................................................................................ 41
Figure 39: Functional block diagram of the DPP-PSD firmware [123]................................................................. 42 Figure 40: Graphical representation of a digitized signal with the DPP-PSD parameters using the Leading-edge discrimination mode. Si corresponds to the signal samples [123]...................................................................... 43 Figure 41: Graphical representation of two detectors operating in coincidence mode..................................... 45 Figure 42: Example of data set in CSV format acquired with Wave mode......................................................... 46 Figure 43: Example of primary data visualization using the code developed with ROOT. The data visualization formats are PSD Vs PID (a), PSD Vs TOF (b), PID Vs TOF (c) and total PID (d). If the TOF is not available, only (a) and (d) are shown................................................................................................................................................
47 Figure 44: Example of PID rejection on 2 MeV neutron applying a cut only on the PSD (a) and on the PSD plus TOF (b)................................................................................................................................................................. 47 Figure 45: AMANDE experimental hall................................................................................................................ 48 Figure 46: Differential cross sections for the monoenergetic neutron production in function of the neutron energy at 0° degree. The numbers marked on the curves indicate the energy of the incident charged particles [126]..................................................................................................................................................................... 49 Figure 47: PTB’s monoenergetic neutrons experimental hall............................................................................. 50 Figure 48: The PTB cyclotron experimental hall [127]........................................................................................ 51 Figure 49: Spectral neutron yield per unit beam charge on the Be target at 0° for proton energies of (a) 17.24 MeV, (b) 19.08 MeV and (c) 20.97 MeV [129].................................................................................................... 52 xxii Figure 50: Gamma uncalibrated PIDs (normalized on the total counts number) for: (a) 22Na and 137Cs, (b) 207Bi and 60Co, (c) 241Am and (d) 12C*........................................................................................................................... 57 Figure 51: Direct measurement experimental set-up simulated with MCNPX-Polimi. Representation of the plane parallel to XY plane (above) and to ZY (down).................................................................................................... 58 Figure 52: Picture of the coincidence measurements experimental set-up....................................................... 59 Figure 53: Comparison between the normalized experimental PID with (w: red full line) and without (w/o: blue full line) the coincidence method for: 207Bi (a), 22Na (b), 60Co (c) and 137Cs (d) sources...................................... 60 Figure 54: Coincidence experimental set-up simulated with MCNPX-Polimi. Representation of the plane
parallel to XY plane (above) and to ZY (down).................................................................................................................
61 Figure 55: LMAX and L1/2 positions on the 137Cs PID....................................................................................... 62 Figure 56: Comparison between the energy resolution (a) and FWHM (b) curves for stilbene (black full line) and BC501A (black dotted line) scintillators. For the energy resolution, the experimental points were obtained using equation (19) and fitted according to equation (3). For the FWHM, points were obtained through FWHM(E) = R ∙ L(E) and the lines are the result of the fitting on these points through equation (21)............................... 63 Figure 57: The normalized and smoothed PID given by a 137Cs (full black square) source and its first order derivate (empty black square), the intersection between the experimental spectrum and the dotted line perpendicular to the lowest point of the Gaussian distribution indicates the CE position evaluation.............. Comparison between 241Am simulated (dashed black line) and experimental calibrated spectra using LMean (red full line) and LLow (blue full line)................................................................................................................ 67
Figure 62: Comparison of the energy calibration curves in the energy range between 0.5 MeV and 4.5 MeV (Left). Comparison between 12C* simulated (dashed black line) and experimental spectra calibrated using LMean (red full line) and LHigh (blue full line) (Right)..................................................................................... 68 Figure 63: Comparison of the stilbene calibration curves obtained without (no Coinc, blue dashed line) and with (Coinc, red full line) the coincidence method of LMean (a) and LLow (b)........................................................ 69 Figure 64: Comparison of the 207Bi experimental spectrum in coincidence mode with the simulated spectra without (a) and whith the stilbene’s energy resolution (b) obtained with MCNPX-Polimi................................ 70 Figure 65: Comparison of the 207Bi experimental spectrum in coincidence mode with the simulated broadened spectra obtained with MCNPX-PoliMi taking into consideration only the events of single scattering.............. 71 Figure 66: Comparison between simulated (black dashed line) and experimental (blue solid line) spectra obtained with the coincidence method for: 22Na (a), 137Cs (b), 60Co (c) and 152Eu (d) sources........................... 73 Figure 67: Comparison between the simulated spectrum and the experimental spectrum calibrated with LMean of the coincidence method (w: blue full line) and without (w/o: red full line) the coincidence method for the 60 Co (a), 137Cs (b), 22Na (c) and 207Bi (d) sources................................................................................................... 74 Figure 68: Calculated gamma ray efficiency curves, according to equation (24), for Ø25.4 × 25.4 mm stilbene with several light-output thresholds. Experimental efficiencies were obtained with a source of 137Cs (black solid squares)................................................. .............................................................................................................. 75 Figure 69: Stilbene gamma ray response matrix between 0.01 and 7.3 MeV energy with 1024 bins between 0 and 8 MeVee....................................................................................................................................................... 76 Figure 70: Photo of the experimental setup for the study of monoenergetic neutrons produced by AMANDE. .............................................................
................
................................................................................................
81 Figure 71: Photo of the experimental setup for the study of monoenergetic neutrons produced at the PTB. 83 xxiii Figure 72: Experimental results obtained with 17 MeV (1) and 5 MeV (2) monoenergetic neutrons. Twodimensional histograms show PID vs PSD (a) and the in the PIDs (b) are shown the effects of cuts applied only on the PSD (blue solid line).................................................................................................................................. 84 Figure 73: Comparison of experimental PIDs for 5 MeV neutrons for direct measurement (solid black line), measurement with shadow cone (solid red line), and measurement without the scattered neutrons (solid blue line)...................................................................................................................................................................... 85 Figure 74: Normalized PIDs for neutrons of: 1.5 MeV (orange full line), 1.004 MeV (blue full line), 0.805 MeV (green full line), 0.688 MeV (red full line), 0.565 MeV (violet full line) and 0.481 MeV (black full line)............ 85 Figure 75: (Left) Calculated FOM as a function of the measured light output for the stilbene. (Right) Comparison of the FOM of two stilbene crystals grown by different methods [96].............................................................. 86 Figure 76: Schematic representation of time of flight applied in AMANDE........................................................ 86 Figure 77: Time of flight distributions for neutrons having 5 MeV (full blue line) and 3.492 MeV (red full line) kinetic energies.................................................................................................................................................... 87 Figure 78: Experimental results obtained with 17 MeV (1) and 5 MeV (2) monoenergetic neutrons. Twodimensional histograms show TOF vs PSD (a) distributions. On the PIDs (b) are shown the effects of cuts applied only on the PSD (blue full line) and PSD plus TOF (red full line)......................................................................... 88 Figure 79: 17 MeV neutron PID (black cross) and its first order derivate (blue cross). The intersection between the experimental spectrum and the red dotted line perpendicular
to
the
lowest point
of the
Gaussian distribution indicat
es
the PE position..................................................................................................................
89 Figure 80: Light-output produced by the stilbene in function of the energy deposited by the protons (full black squares). Proton data have been fitted using equation (12) (full blue line) and compared with results from other stilbene crystals (full black line) [54] (full red line) [84] (full green line) [144] (full yellow line) [145]............ 90 Figure 81: Comparison between normalized experimental spectra and simulated spectra broadened with a constant value aver the all spectrum for neutron of 17 MeV (a), 5 MeV (b), 2.5 MeV (c) and 0.805 MeV (d). 91 Figure 82: Neutron energy resolution data fitted with equation (3)..................................................................
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tel-04602906-HADJ%20KALI_2004-12-08.txt_2
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Open Science
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Application de la simulation moléculaire pour le calcul des équilibres liquide-vapeur des nitriles et pour la prédiction des azéotropes. Génie des procédés. Institut National Polytechnique (Toulouse), 2004. Français. ⟨NNT : 2004INPT024G⟩. ⟨tel-04602906⟩
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L’ensemble isobare isotherme NPT
L’ensemble NPT est largement utilisé dans les simulations de type Monte Carlo notamment parce qu’il permet une comparaison directe avec des résultats issus d’expérimentations réelles qui sont fréquemment réalisées à température et pression contrôlées. La fonction de partition ∆NPT de cet ensemble est donnée par l’expression : (
( )) ∆ NPT = ∑∑ exp − β E j + PV = ∑ exp(− β PV ) × Q NVT j V (II-28)
V Notons que la quantité qui apparaît dans l’exponentielle de cette fonction, lorsqu’on évalue la moyenne, donne l’enthalpie thermodynamique H = E + PV. Dans cet ensemble, le volume V rejoint la liste des quantités microscopiques (les coordonnées et les vitesses) décrivant les états quantiques du système. Le potentiel thermodynamique associé à l’ensemble NPT est l’énergie libre G. En effet, on peut montrer facilement que :
G
=
−kBT
ln ∆ NPT
= H −
T S iv. (II-29)
L’ensemble grand canonique μVT
Pour quelques applications, il est commode de travailler dans un ensemble où les systèmes peuvent échanger non seulement de l’énergie mais aussi de la matière (exemple au chapitre IV). Un ensemble similaire est appelé ensemble grand canonique qui correspond à un nombre important de systèmes ouverts de volume constant, en équilibre interne et pouvant échanger de l’énergie et de la matière (les molécules) avec le milieu extérieur. Pour décrire l’état thermodynamique d’un tel ensemble, il est pratique d’utiliser comme variables caractéristiques indépendantes la température T, le volume V et les potentiels chimiques μ1, μ2,... des constituants 1, 2,...du système. La fonction de partition est donnée alors par la relation : ( ( )) Q μVT = ∑∑ exp − β E j − μN = ∑ exp(βμN ) × Q NVT j N (II-30) N Le potentiel thermodynamique associé à cet ensemble est le produit -PV étant donné que : − PV = −kBT ln Q μVT (II-31) Il est possible de construire beaucoup d’autres ensembles, mais ils ne sont pas tous intéressants pour la simulation moléculaire. En effet, considérons l’exemple de l’ensemble statistique μPT où les paramètres constants sont toutes des variables intensives ; dans ce cas, les quantités extensives correspondantes sont sujettes à des fluctuations, ceci signifie que la taille des systèmes peut augmenter sans limite. D’autre part, le vecteur potentiel chimique μ, la pression P et la température T du système sont reliés par une équation d’état, et donc, même si l’expression de cette équation était connue, ces trois variables ne sont pas II-10 indépendantes. Pour toutes ces raisons, la simulation dans un ensemble μPT ou d’autres ensembles présentant les mêmes caractéristiques n’est pas pratique. C’est pourquoi, la majorité des ensembles usuels en simulation moléculaire nécessite la spécification, d’au moins, une variable extensive (généralement le volume V ou le nombre de particule du système N) afin de pouvoir contrôler la taille du système étudié. Notons enfin que les couples typiques de variables intensives et leurs extensives conjugués sont : (T, E), (P, V) et (μ, N) ce qui nous permet de retrouver, par combinaison directe, le choix des variables caractéristiques des ensembles classiques. c. Relation entre équation d’état et thermodynamique statistique
La base théorique sur laquelle repose toute la thermodynamique est la mécanique statistique. Par conséquent, la mécanique statistique a été utilisée comme point de départ pour le développement de plusieurs équations d’état succinctement résumées comme étant de la forme f(P,V,T) = 0. En effet, comme nous l’avons décrit un peu plus loin, c’est dans la fonction de partition d’un ensemble donné que se concentrent toutes les informations concernant le système étudié. De ce fait, lorsqu’on fixe la température T, le volume du système V et le nombre de particules N comme variables caractéristiques indépendantes (c'est-à-dire lorsqu’on travaille dans l’ensemble canonique), toutes les informations décrivant le système sont contenues dans la fonction de partition canonique ayant pour expression : Q NVT = ∑ e − E j ( N,V ) kB T (II-19) j Le potentiel thermodynamique caractéristique de cet ensemble est l’énergie libre d’Helmholtz, dont on rappelle l’expression : F (N, V, T ) = −k B T ln Q NVT (II-26) Ainsi, une fois la fonction de partition QNVT ou, similairement, l’énergie libre du système F connue en fonction des variables caractéristiques N, V et T, toutes les autres propriétés thermodynamiques seront accessibles [ Sandler, 1994 ] ; en particulier, l’équation d’état sera donnée par la relation : ∂ ln Q NVT ∂F = − P (N,V, T ) = k B T ∂V T, N T,N ∂V (II-32) D’autre part, l’expression de la fonction de partition peut être précisée en séparant les contributions cinétique et potentielle à l’énergie du système et en remplaçant les sommations discrètes par des intégrales, ce qui permet d’écrire la fonction de partition sous la forme : Q NVT = q i (T )N Z NVT N! (II-33) II-11 Où q i (T ) représente la fonction de partition moléculaire qui traduit les contributions vibratoire, rotationnelle, électronique et nucléaire. Pour de petites molécules, q i (T ) n’est fonction que de la température et donc n’affecte pas l’équation d’état volumétrique. L’intégrale de configuration ZNVT, Z NVT = ∫ L∫ e −U (r1, r2,Kr N ) kT dr1dr 2 L dr N (II-34) est le facteur de Boltzmann de l’énergie d’interaction U intégrée sur toutes les positions des particules. Si nous considérons un système idéal ou les particules n’interagissent pas entre elles, U prendra la valeur 0 et on obtiendra immédiatement Z = V N, ce qui, introduit dans l’équation (II-32), donne l’équation d’état d’un gaz parfait :
P ∂ qi (T )N N ∂ ln V ∂ ln Q ln V = N = = = NV k BT ∂V ! V N V ∂ ∂ T, N T,N T, N (II-
35) En multipliant cette équation par le nombre d’Avogadro NA, on obtient l’expression classique de l’équation d’état d’un gaz parfait : PV =
nRT
ou
P
=
ρ k BT
équation d’état du gaz parfait (II-36) Où ρ est la densité moléculaire. La constante des gaz parfaits R est égale au produit de la constante de Boltzmann kB=1,38066.10-23 J/K et du nombre d’Avogadro NA=6,023.10+23 entitées. Il s’agit d’une illustration remarquable de la façon dont une hypothèse microscopique très simple (pas d’énergie d’interaction) permet de dériver une équation macroscopique importante. Ce comportement est approché par tous les gaz réels aux faibles densités. A de plus fortes densités, les particules se rapprochent et interagissent de façon non négligeable. Dans ce cas, l’intégrale de configuration ZNVT n’est plus seulement égale à VN. Deux approches peuvent alors être employées pour caractériser le comportement non idéal des fluides loin de la zone des faibles densités
: -
L’é
quation
d’état
du
viriel décrit les déviations du comportement idéal par un développement limité en série de Taylor de la densité moléculaire : P = ρ + B(T )ρ 2 + C (T )ρ 3 +... k BT équation du viriel (II-37)
Dans cette équation, B(T) est le second coefficient du viriel et est associé aux interactions entre paire de molécules, C(T) est le troisième coefficient du viriel et est associé aux interactions entre triplets de molécules, etc. - A l’exception des cas de gaz rare ou présentant une infime déviation par rapport au comportement idéal, l’intégrale de configuration des fluides ne peut être évaluée exactement. II-12 Du fait que la mécanique statistique ne fournie pas de solution exacte pour les fluides réels, plusieurs méthodes d’approximations et des modèles d’interaction sont employés dont la théorie des états correspondants, la théorie de perturbation et d’autres méthodes d’approximation de la fonction de partition comme la théorie de Van der Waals généralisée. Ils permettent alors de dériver des équations d’états spécifiques, par exemple l’équation de Van der Waals.
B. Simulation moléculaire : Techniques d’échantillonnage 1. Introduction
La simulation moléculaire est basée sur des concepts de thermodynamique statistique et de mécanique moléculaire. La qualité des résultats de simulation moléculaire dépend du type de système modèle employé, de l’utilisation de modèles de mécanique moléculaire décrivant avec précision les interactions inter et intramoléculaires et aussi de la qualité de l’échantillonnage statistique des configurations du système modèle. L’échantillonnage de l’ensemble statistique des configurations du système est réalisé principalement au moyen de deux techniques : la Dynamique Moléculaire et la méthode de Monte Carlo. Ces deux techniques sont présentées dans ce sous-chapitre. 2. La méthode de Monte Carlo a. Introduction
Une fois que la modélisation relative à un système physique donné a été choisie, la deuxième étape du travail consiste à déterminer les propriétés statistiques du modèle en effectuant une simulation. Si l'on s'intéresse aux propriétés statiques du modèle, nous avons vu au sous-chapitre précédent que le calcul de la fonction de partition du système se ramène au calcul d'une intégrale ou d'une somme discrète de configurations de la forme : Z = ∑ exp(− βU i ) (II-38) i Où i est un indice parcourant l'ensemble des configurations accessibles au système. En prenant N=10 points pour chaque coordonnée d'espace et avec M=100 particules évoluant dans un espace à D=3 dimensions, le nombre de configurations accessibles est alors égal à ND.M=10300, ce qui rend impossible le calcul complet de la somme dans l'équation (II-38). Il est donc nécessaire de disposer de méthodes spécifiques pour évaluer les intégrales multidimensionnelles. Une technique utilisée est la méthode dite de Monte Carlo avec un algorithme d' « échantillonnage d'importance ». L’expression « méthode Monte Carlo » est très générale et concerne toutes les méthodes stochastiques basées sur l’utilisation des nombres aléatoires et des probabilités statistiques pour la résolution des problèmes. Elles sont utilisées dans des domaines aussi variés que l’économie, la physique nucléaire ou la régulation de la circulation dans le trafic. II-13 L’utilisation de la méthode Monte Carlo pour modéliser les problèmes de physique nous permet d’étudier des systèmes complexes en générant aléatoirement un grand nombre de configurations parmi l’infinité des configurations que peut occuper un système. b
. Les générateurs de nombres aléatoires
Les générateurs de nombres aléatoires sont au cœur de toutes les simulations de Monte Carlo, non seulement pour générer de nouvelles configurations mais aussi pour décider de l’acceptation ou du rejet de n’importe quel mouvement effectué. On les retrouve également dans l’initialisation des simulations de Dynamique Moléculaire présentées après pour attribuer les vitesses initiales aux différentes molécules du système. Le prototype d'un générateur de nombres aléatoires doit fournir des nombres issus d'une distribution uniforme. Cependant, pour que ces nombres soient vraiment aléatoires, il est nécessaire de satisfaire une infinité de critères : la moyenne, la variance, mais aussi tous les moments de la distribution doivent être ceux d'une distribution uniforme. De plus, les suites de nombres doivent être sans corrélation entre elles. Comme les nombres sont représentés par un nombre fini d'octets en informatique, les générateurs sont forcément périodiques. Un critère nécessaire mais non suffisant est d'avoir une période très élevée. Dans les premiers temps de l'informatique, les générateurs utilisaient une représentation des nombres sur 8 bits, impliquant des périodes très courtes et des résultats systématiquement faux pour les simulations Monte Carlo. C’est pourquoi, lors des simulations nécessitant un grand nombre de tirages, il est indispensable de s'assurer que la période du générateur reste très supérieure au nombre de tirages. Mais d'autres qualités sont nécessaires, comme l'absence de corrélation entre les séquences de nombres ; l'initialisation correcte du générateur reste un point encore trop souvent négligé. Il existe deux grands types d'algorithme pour obtenir des générateurs de nombres aléatoires. Le premier type est basé sur la congruence linéaire : x n +1 = (ax n + c ) × mod(m ) (II-39) mod étant la fonction modulo. Ce type de relation génère une suite « pseudo-aléatoire » de nombres entiers compris entre 0 et (m-1). m donne la période du générateur. Par conséquent, il suffit de diviser xn+1 par m pour obtenir des nombres aléatoires compris entre 0 et 1, puisque xn+1 ne peut excéder la valeur de m. Parmi les générateurs utilisant cette relation, on trouve les fonctions randu d'IBM, ranf du Cray, drand48 sur les machines Unix, ran de Numerical Recipes (Knuth), etc. Les périodes de ces suites de nombres vont de 229 (randu IBM) à 248 (ranf). La deuxième classe de générateurs est basée sur le déplacement de registre à travers l'opération « ou exclusif ». Un exemple est donné par le générateur de Kirkpatrick et Stoll.
II-14 x n = x n −103 ⊕ x n − 250 (II-40)
Sa période est grande, 2250, mais il nécessite de stocker 250 mots. Le générateur avec la période la plus grande est sans doute celui de Matsumoto et Nishimura connu sous le nom MT19937 (Mersenne Twister generator). Sa période est de 106000! Il utilise 624 mots par générateur et il est équidistribué dans 623 dimensions! c.
Exemple illustratif
Pour illustrer l’application d’une méthode d’échantillonnage basée sur des nombres aléatoires dans le but de calculer une propriété macroscopique, prenons l’exemple du calcul de la valeur de π. La Figure II-1.a représente un cercle unitaire inscrit sur un carré. On pourrait examiner ce problème en terme du cercle complet et du carré, mais il est plus facile de traiter le problème en considérant uniquement le quart, comme le montre la Figure II-1.b.
y 1 1 0 1 x Figure II-1 : Calcul de π par la méthode Monte Carlo.
Une personne tente de placer un grand nombre de fois un objet dans le quart du cercle. A chaque tentative, les coordonnées (x,y) de l’objet sont déterminées par deux nombres aléatoires indépendants choisis à partir d’une distribution uniforme entre 0 et 1. La distance r entre chaque objet et l’origine est calculée par la relation : r = x 2 + y 2. Si cette distance est inférieure ou égale à 1, l’objet a atterri sur la zone hachurée et la tentative est comptée, sinon, l’essai est rejeté. Si on note ‘n’ le nombre d’essais inscrits à l’intérieur du quart de cercle sur ‘N’ tentatives exécutée, la valeur de π peut être calculée par l’expression : π= 4 × la surface du quart de cercle hachurée 4 × n = la surface du carré N (II-41) L’application d’une technique de Monte Carlo faisant appel à un générateur de nombres aléatoires pour cet exemple consiste à la génération de ‘2N’ nombres aléatoires compris entre 0 et 1 à l’intérieur d’une distribution uniforme qui seront affectés au couple de coordonnées (x,y). II-15
Cet exemple est facile à comprendre ; en simulation moléculaire, par contre, nous sommes face à des problèmes d’évaluation de moyennes et d’intégrales beaucoup plus complexes pour un nombre très élevé de configurations. C’est pourquoi l’application des méthodes de Monte Carlo pour la simulation moléculaire a fait l’objet d’un travail particulier aboutissant à des techniques spécifiques (l’algorithme de Métropolis, les biais statistiques,...) dont nous présentons, ci-dessous, les principes théoriques.
d. Echantillonnage uniforme
Tout d'abord considérons l'exemple le plus simple, celui d'une intégrale unidimensionnelle : b I = ∫ f (x )dx (II-42) a Cette intégrale peut être réécrite sous la forme : I = (b − a ) f (x ) Où (II-43) f (x ) représente la moyenne de la fonction sur l’intervalle [a,b]. En choisissant aléatoirement et uniformément N points le long de l'intervalle [a,b], et en calculant la valeur de la fonction pour chacun de ces points, on obtient une estimation de l'intégrale par l'expression : I≈ (b − a ) N f ( x ) ∑ N (II-44) i i =1 La Figure II-2 illustre le principe de l’échantillonnage uniforme.
f f a b Echantillonnage uniforme état f a b Echantillonnage aléatoire état a b état Echantillonnage préférentiel
Figure II-2. Illustration des échantillonnages uniforme, aléatoire et préférentiel
Pour appliquer cette approche à l’estimation de la valeur de π, il suffit de considérer l’équation du cercle dans le domaine de travail, c’est à dire pour x compris entre a = 0 et b = 1 : ( f ( x) = 1 − x 2 ) 1/ 2 (II-45) Une estimation de la valeur de π par cette fonction donne la valeur de 3,14169, pour 107 tentatives, soit quatre chiffres significatifs. II-16 Considérons l’exemple d’évaluation de l’intégrale de configuration ZNVT pour un système de N molécules disposées dans un cube d’arête L [ Allen et Tildesley, 1987 ]. Z NVT = ∫ exp( − βU )dr (II-46) Pour approcher cette intégrale par cette méthode à l’aide d’une distribution uniforme, on procède selon les étapes suivantes : - On génère 3N nombres aléatoires dans le domaine –L/2 et +L/2. - Pris par triplet, ces nombres spécifient les coordonnées des N molécules à l’intérieur du cube. - On calcule l’énergie du système grâce à un champ de force qui est fonction des positions des différentes particules dans la configuration obtenue. - Cette procédure est répétée Nmax fois (Nmax >> 106), ce qui permet d’estimer l’intégrale de configuration par la relation : Z NVT = V N max ∑ exp( − βU ( n )) N max n =1 (II-47) Hélas, cette évaluation de l’intégrale de configuration n’est pas envisageable pour le calcul des propriétés thermodynamiques d’un système moins idéal vu le très grand nombre de configurations d’un système qui ont une probabilité d’existence ou encore un facteur de Boltzmann quasi nul à cause d’une énergie trop élevée du fait du recouvrement énergétique entre les particules. Ces configurations peu probables sont innombrables et donc longues à calculer et pourtant n’interviennent pas ou peu dans le calcul des grandeurs moyennes. La convergence de cette méthode peut être estimée en calculant la variance σ 2 de la somme I. Or, on a :
σ2 = ( 1 N N ∑∑ f (x i ) − f (x i ) N 2 i =1 j =1 ) ( f (x ) − f (x ) ) j j (
II
-
48) Les points étant choisis indépendamment, les termes croisés qui qualifient la corrélation s'annulent, et on obtient
σ2 = ( 1 N 2 f (x i )2 − f (x i ) ∑ N i =1 )
(II-49) La dépendance en 1/N donne une convergence a priori assez lente, mais il n'y a pas de modifications simples pour obtenir une convergence plus rapide. On peut à l'inverse modifier de façon importante l'écart type. Pour cela, il paraît clair que si la fonction f(x) ne prend des valeurs significatives que sur des petites régions de l’intervalle [a,b], il est inutile de calculer la fonction en des points où sa valeur est très faible.
II
e
. Sample mean integration : échantillonage aléatoire
Si par contre, on travaille avec une distribution aléatoire non uniforme (Figure II-2) avec un poids ρ(x), l'intégrale se réécrit : f (x ) ρ (x )dx ( ) ρ x a b I =∫ (II-50) Si ρ(x) est toujours positif, on peut définir du = ρ (x ) dx avec u (a ) = a et u (b ) = b, et on a alors une estimation de l'intégrale qui est donnée par f (x (u )) du ρ (x (u )) a (II-51) (b − a ) N f (x (u i )) ∑ N i =1
ρ (x (u i )) (II-
52)
b I =∫ I≈
Avec la distribution de poids ρ(x), de manière similaire, la variance de l'estimation devient alors 1 N
f (x (u i )) σ = ∑ N i =1 ρ (x (u i )) 2 2 f (x (u i )) − ρ (x (u i )) 2
(II-53) En choisissant la fonction de poids ρ(x) proportionnelle à la fonction f(x), la variance s'annule. Cette merveilleuse astuce n'est possible que dans un problème à une dimension. En dimension supérieure, le changement de variables dans une intégrale multiple fait intervenir la valeur absolue d'un jacobien et on ne peut donc pas trouver de manière intuitive le changement de variable à effectuer pour obtenir une fonction de poids satisfaisante. Pour des intégrations unidimensionnelles simples, la technique de Monte Carlo n’est pas compétitive avec les méthodes numériques de type Simpson par exemple bien connue des mathématiciens pour calculer des intégrales simples. Cependant, pour les intégrales de thermodynamique statistique, de dimension 6N pour un système à N particules, la méthode « sample mean integration » avec un choix approprié de la fonction de distribution de densité ρ(x) est la seule à fournir une solution acceptable. Une solution au problème d’évaluation de l’intégrale de configuration ZNVT, introduite dans le paragraphe précédent, consiste à ne générer que les configurations les plus stables et qui participent donc le plus à l’évaluation des grandeurs thermodynamiques moyennes. C’est la stratégie adoptée par la méthode « échantillonnage préférentiel » ( importance simpling ) introduite par Metropolis [ Metropolis et al., 1953 ] (Figure II-2) et qui est employée pour la prédiction des équilibres liquide - vapeur.
II-18 f.
Chaîne de Markov pour échantillonner le système à l'équilibre
Reprenons notre problème de mécanique statistique : nous sommes intéressés le plus souvent par le calcul de la moyenne d'une grandeur A et non pas directement par la fonction de partition. Cette moyenne s’écrit dans le cas de l’ensemble canonique sous la forme : A = ∑ A exp(− βU ) i i i (II-54) Z NVT Où i est un indice parcourant l’ensemble des configurations accessibles au système. On rappelle, d’autre part, que : pi = exp(− β U i ) Z NVT (II-55) définit la probabilité d'avoir la configuration i à l'équilibre (pi est toujours positif et ∑ p = 1 ). Si l'on i i était capable de générer des configurations avec un même poids peq, la moyenne de A serait estimée par : A ≈ 1 N ∑ Ai N i (II-56) Où N est le nombre de points calculés ; on serait alors ramené au calcul de la section précédente. L'astuce imaginée par Metropolis, Rosenbluth et Teller en 1953 consiste à générer une dynamique stochastique Markovienne stationnaire, entre configurations successives, qui converge vers la distribution d'équilibre peq. Avant d'expliciter ce point, nous allons introduire quelques définitions. Considérant l'ensemble des configurations i, on introduit un temps t prenant les valeurs discrètes associées au comptage des itérations dans la simulation. Ce temps n'a pas de relation directe avec le temps réel du système. On appelle p (i, t ) la probabilité du système d'être dans la configuration i au temps t. Reprenons maintenant les termes de la dynamique choisie : dynamique stochastique signifie que le passage d'une configuration à une autre est le choix d'une procédure aléatoire et Markovien signifie que la probabilité d'aller vers une configuration j à l'instant t+1, sachant que le système était dans la configuration i à l'instant t, ne dépend pas des configurations du système pour des instants antérieurs (mémoire limitée à l'instant t); cette probabilité conditionnelle est notée w(i → j ). L'équation d'évolution du système est alors donnée par l'équation maîtresse suivante :
p(i, t + 1) = p(i, t ) + ∑ (w( j → i ) p( j, t ) − w(i → j ) p(i, t )) (II-57) j II-19
Cette équation traduit le bilan suivant : à l'instant t+1, la probabilité du système d'être dans l'état i est égale à celle de l'instant précédent, augmentée par la possibilité que le système qui se trouve dans n'importe quelle autre configuration puisse aller dans l'état i et diminuée par la possibilité que le système qui se trouvait dans l'état i puisse aller vers n'importe quelle autre configuration. A l’instant initial t = 0, le système est placé dans une configuration initiale i0, ce qui signifie que, quel que soit le choix de cette configuration, le système ne satisfait pas P(i ) = ρ i qui est la condition recherchée. ρ i étant la densité de probabilité de la configuration i. Afin que le système converge vers l'équilibre, avec l'équation d'évolution (II-57), on obtient l'ensemble des conditions suivantes :
∑ w( j → i ) ρ = ρ ∑ w(i → j ) j (II-58) i j j
Une solution simple de ce système d'équations est donnée par : w( j → i )ρ j = w(i → j )ρi micro-réversibilité (II-59) Cette relation, (II-59), est connue sous le nom de micro-réversibilité ou de bilan détaillé. Elle exprime le fait que, dans l'état stationnaire, (ou état d'équilibre si le processus n'a pas engendré une brisure d'ergodicité), la probabilité que le système puisse aller d'un état d'équilibre i vers un état j est la même que celle d'aller d'un état d'équilibre j vers un état i. Ajoutons que cette condition n'est qu'une condition suffisante, car nous n'avons pas prouvé simultanément que la solution du système d'équations (II-58) est unique et que l'équation (II-59) est la meilleure solution. Pour des raisons pratiques, la quasi-totalité des algorithmes de Monte Carlo reposent sur cette solution. L'équation (II-59) peut être aisément réécrite sous la forme : w(i → j ) ρ j = = exp − β U j − U i w( j → i ) ρi ( ( )) (II-60) Cela implique que les inconnues w(i → j ) que l'on cherche à déterminer ne dépendent pas de l’intégrale de configuration ZNVT si difficile à évaluer, mais uniquement du facteur de Boltzmann relié à l’énergie de chaque état qui peut être calculée. Algorithme de Metropolis
Le choix du processus Markovien stationnaire pour satisfaire l'ensemble des équations (II-59) est l'une des solutions. Afin d'obtenir des solutions des équations (II-59) ou, en d'autres termes d'obtenir la matrice de transition (w(i → j )), notons que le processus stochastique élémentaire dans un algorithme de Monte Carlo est la succession de deux étapes : II-20 - A partir d'une configuration i, on tire au hasard une configuration j, avec une probabilité « a priori » d’essayer d’aller de l’état i à l’état j α (i → j ). - Cette nouvelle configuration est acceptée avec une probabilité d’acceptation π (i → j ). Ainsi, on a w(i → j ) = α (i → j ) ⋅ π (i →
j
) (II-61) Dans l'algorithme original de Metropolis (et dans la plupart des algorithmes Monte Carlo), on choisit une fonction symmétrique α (i → j ) = α ( j → i ). Dans ce cas, les équations (II-60) s’expriment comme : π (i → j )
= exp(− β (U j − U i )) π ( j → i)
(II-62) La solution choisie par Metropolis et al. [ 1953 ] est :
π (i → j ) = 1 si U j ≤ Ui π (i → j ) = exp(− β (U j − U i )) si U j > Ui (II-63)
Ainsi, dans la méthode de Metropolis la nouvelle configuration j est toujours acceptée si son énergie est plus faible que celle de l’état original i. Sinon, c’est à dire si l’énergie de cette nouvelle configuration est plus grande, le déplacement est accepté suivant l’équation (II-63) en comparant le facteur de Boltzmann ( ) relatif à la transition exp(− β∆U i→ j ) (avec : ∆U i→ j = U j − U i ) à un nombre aléatoire compris entre 0 et 1 : si le facteur de Boltzmann est supérieur au nombre aléatoire, la nouvelle configuration est acceptée, sinon, le déplacement est refusé et le système restera à son état initial. Ceci revient à dire qu’on accepte n’importe quel déplacement avec la probabilité : π (i → j ) = min (1, exp(− β∆U i→ j )) (II-64) exp(− β U ) Rejeter ξ1 Toujours accepter Accepter ξ2 0 ∆U (i → j ) ∆U
Figure II-3 : Critère d’acceptation de la méthode de Metropolis II-21
La Figure II-3 illustre le critère d’acceptation d’une transition de l’état i à l’état j par la méthode de Metropolis où ξi est un nombre aléatoire compris entre 0 et 1. Cependant, le calcul d'une moyenne ne peut commencer que lorsque le système a atteint l'équilibre, c'est à dire quand p ≈ peq. Ainsi dans une simulation de Monte Carlo, il y a généralement deux périodes : la première, où partant d'une configuration initiale, on réalise une dynamique afin d'amener le système près de l'équilibre ; la seconde période, où le système évolue au voisinage de l'équilibre, et où le calcul des moyennes est réalisé. En l'absence de critère précis, la durée de la première période n'est pas facilement prévisible. Une des techniques consisterait à suivre l'évolution de l’énergie instantanée du système et à considérer que l'équilibre est atteint lorsque l'énergie se stabilise autour d'une valeur quasi-stationnaire.
3. Dynamique moléculaire a. Introduction
La limitation intrinsèque de la simulation de Monte Carlo provient du fait qu'on ne considère pas la dynamique ``réelle'' du système. Pour des systèmes continus définis à partir d'un Hamiltonien classique, il est possible de résoudre les équations du mouvement de la mécanique newtonienne pour l'ensemble des particules. Cette méthode offre le moyen de calculer précisément à partir de corrélations temporelles les propriétés dynamiques du système à l'équilibre, grandeurs qui sont accessibles expérimentalement par diffusion de la lumière ou des neutrons. Cette méthode permet aussi de calculer à partir de corrélations spatiales les grandeurs statiques d'équilibre comme dans une simulation Monte Carlo afin d'être comparées directement à l'expérience. Pour des systèmes atomiques en dehors de la région critique, cela est suffisant, mais ça se révèle limitant pour des édifices moléculaires plus complexes (molécules biologiques). En se basant sur un potentiel de Lennard-Jones pour décrire les interactions énergétiques, le temps obtenu à partir d'une analyse dimensionnelle des paramètres microscopiques du système est : τ =σ m ε (II-65) m étant la masse d'une particule, σ son diamètre, et ε l'échelle d'énergie du potentiel d'interaction. Le temps τ représente le temps pour un atome de se déplacer sur une distance égale à sa taille avec une vitesse égale à la vitesse moyenne dans le fluide. Par exemple, pour l'argon, on a σ=3 Å, m=6.63×10-23 kg et ε=1.64×10-20J, ce qui donne τ=2.8×10-14s. Pour une intégration numérique des équations du mouvement, le pas d'intégration du temps doit rester une fraction du temps τ, typiquement ∆t = 10-15 s, voire plus petite. Le nombre de pas que l'on peut réaliser en dynamique est typiquement de l'ordre de 105 à 107, ce qui nous amène à pouvoir suivre un phénomène au maximum sur un intervalle de temps qui va II-22 jusqu'à 10-8 s. Pour de nombreux systèmes atomiques, les temps de relaxation des phénomènes sont très inférieurs à 10-8 s et la Dynamique Moléculaire est un bon outil d’étude.
. Équations du mouvement
Au cours d’une simulation de Dynamique Moléculaire, on détermine l’évolution du système au cours du temps. Soit Xobs une propriété macroscopique d’un système de N particules que l’on souhaite étudier. A un instant t donné, le système est dans un état correspondant à un point de l’espace des phases caractérisé par 6N coordonnées : 3N coordonnées de position et 3N coordonnées de quantité de mouvement. La valeur instantanée X(t) de Xobs peut être exprimée en fonction de ces 6N coordonnées. Le principe de la Dynamique Moléculaire est de résoudre numériquement les équations de mouvement du système étudié. On obtient alors une trajectoire dans l’espace des phases. Pour chaque point de cette trajectoire, il est possible de calculer la valeur de X(t) et la grandeur Xobs mesurable expérimentalement peut être assimilée à la moyenne temporelle de X(t) : X obs = X (t ) = lim τ →∞ 1 τ τ ∫0 X (t )dt (II-1) τ étant le temps d’observation. La Dynamique Moléculaire est une technique utilisée pour résoudre l’équation classique de mouvement de newton pour un système de N particules soumises à un champ de potentiel. Pour une particule de masse mi constante se déplaçant avec une vitesse vi et soumise à des forces fi, l’équation simplifiée s’écrit : dv i ∑ f = m dt i (II-66) i Les forces fi agissant sur la particule de masse mi sont égales à la dérivée du potentiel U(r) agissant sur la particule. Dans la direction x : f x (r ) = − ∂U ( r ) x dU (r ) = − ∂x r dr (II-67) σ 12 σ 6 r − r , la Ainsi, soumise à un potentiel de Lennard-Jones 12-6 de la forme : U VdW = 4ε ⋅ 12 6 48 ⋅ ε ⋅ x σ 1 σ ⋅ − force sera égale à f x,VdW = r 2 r r2 Pour simuler un milieu infini, on utilise dans une simulation des conditions aux limites périodiques. Le calcul de la force interagissant entre deux particules i et j se fait souvent entre l'image de j la plus proche de i (voir le paragraphe sur les techniques de calcul). II-23
Comme dans le cas d’une simulation de Monte Carlo, le calcul de la force agissant sur la particule i nécessite a priori le calcul de (N-1) forces élémentaires provenant des autres particules. L'utilisation d'un potentiel tronqué dans l’espace permet alors de limiter le calcul de la force aux particules entourant la particule i dans une sphère dont le rayon est celui de la troncature du potentiel.
c. Discrétisation. Algorithme de Verlet
Pour intégrer numériquement des équations différentielles, il est nécessaire de les discrétiser en temps. Une grande variété de choix est possible a priori, mais comme nous allons le voir par la suite, il est important que l'énergie du système soit conservée au cours du temps (l'ensemble statistique est ici l’ensemble microcanonique, NVE). L'algorithme proposé par Verlet est historiquement l'un des premiers introduit et il reste encore l'un des plus utilisés actuellement. Pour des raisons de simplicité, nous considérons un système constitué de N particules identiques et nous appelons r, un vecteur à 3N composantes: r = (r1, r2,..., rN), où ri désigne le vecteur position de la particule i. L'équation d'évolution du système peut s'écrire formellement comme : m d 2r = f (r (t )) dt 2 (II-68) En faisant un développement de Taylor, on a :
r (t + ∆t ) = r (t ) + v(t )∆t + 3 f (r (t )) (∆t )2 + d 3r (∆t )3 + O (∆t )4 2m dt ) (II-69) 3 f (r (t )) (∆t )2 − d 3r (∆t )3 + O (∆t )4 2m dt ) (II-70) ( et de manière similaire, r (t − ∆t ) = r (t ) − v(t )∆t + ( En
sommant ces
deux équations, on obtient : r (t + ∆t ) + r (t − ∆t ) = 2r (t ) + f (r (t )) (∆t )2 + O (∆t )4 2m ( ) (II-71)
Le calcul de la nouvelle position est donc effectué avec une précision de l'ordre de (∆t)4. Cet algorithme a l’avantage de ne pas utiliser les vitesses des particules pour calculer les nouvelles positions, vitesses qui requiert de calculer la dérivée des trajectoires r(t). On peut toutefois déterminer les vitesses a posteriori de la manière suivante :
v(t ) = r (t + ∆t ) − r (t − ∆t ) 2 + O (∆t ) 2 ∆t ( ) (II-
La qualité d'une simulation de Dynamique Moléculaire est liée à la qualité de l'algorithme utilisé et à ses propriétés. La rapidité de l'exécution du programme peut aussi être déterminante. Notons que l'essentiel II-24 du temps de calcul dans une Dynamique Moléculaire est consommé dans le calcul des forces, ce qui signifie que le coût du calcul des nouvelles positions est marginal. La précision du calcul pour l'algorithme de Verlet est approximativement donnée par ∆t 4 × N t (II-73) Où Nt est le nombre de pas d’intégration de la simulation. Le temps maximal écoulé dans la simulation est donné par ∆t × N t. Il pourrait sembler intéressant d'utiliser un algorithme faisant intervenir des dérivées des coordonnées à des ordres plus élevés mais dans ce cas, la quantité d'information à garder en mémoire augmente rapidement. Ainsi, l’algorithme de Verlet est particulièrement compact, ne nécessitant que 9 vecteurs de N éléments (r x, r y, r z, r& x, r& y, r& z, et les nouvelles forces Fx, Fy, Fz), là où un algorithme type Gear basé sur des équations analogues à l’équation (II-69) en nécessite 15 (r x, r y, r z, r& x, r& y, r& z, &r& x, &r& y, &r& z, &r&& x, &r&& y, &r&& z et les nouvelles accélérations (3N) et nouvelles forces (3N)). Les algorithmes d'ordres plus élevés, comme celui de Gear, ont tendance à fournir une dynamique aux temps courts de meilleure qualité, mais l'énergie totale du système tend à ne pas rester constante aux temps longs. L'algorithme de Verlet possède au contraire la vertu de conduire à une faible dérive de l’énergie totale aux temps longs. Or étudier le plus longtemps la dynamique du système est précisément ce que l’on cherche avec la Dynamique Moléculaire. Une symétrie particulièrement importante, contenue dans les équations de Newton du système, est la symétrie par renversement du temps. Il est important de noter que l'algorithme de Verlet satisfait cette symétrie. En effet, en changeant ∆t → −∆t, l'équation (II-71) reste inchangée. La conséquence de cette propriété est que si, à un instant t de la simulation, on inverse le cours du temps, la trajectoire de la Dynamique Moléculaire revient sur ses pas. En pratique, les erreurs d'arrondi accumulés dans la simulation limitent la réversibilité quand le nombre de pas d’intégration devient important. En revanche, on peut, en utilisant cette propriété de l'algorithme, tester l'importance des erreurs d'arrondi, en inversant le temps dans la simulation pour des temps de plus en plus grands. Signalons un algorithme, apparenté à celui de Verlet, connu sous le nom d'algorithme « Leap frog » (algorithme saute-mouton) basé sur le principe suivant : les vitesses sont calculées pour des intervalles de temps demi-entiers et les positions sont obtenues pour les intervalles de temps entiers. Si on définit les vitesses pour les temps t + ∆t 2 et t − ∆t 2 : v(t +
∆
t 2) =
r (t + ∆t ) − r (t ) ∆t (II-74) v(t − ∆t 2 ) = r (t ) − r (t − ∆t ) ∆t (II
-75) II-25 On obtient :
r
(
t + ∆t ) = r (t ) + v(t + ∆t 2 ) × ∆t (II-76)
Et, de manière similaire, r (t
− ∆t
) = r (t ) − v(t + ∆t 2 ) × ∆t (II-77) En utilisant l'équation (II-71), on obtient la relation : v(t + ∆t 2) = v(t − ∆t 2 ) + f (t ) ∆t + O (∆t )3 2m ( ) (II-78) Puisqu'il est aussi basé sur l'équation (II-71), cet algorithme est identique à l'algorithme de Verlet en ce qui concerne le calcul des trajectoires (les valeurs des vitesses aux temps demi-entiers n'apparaissent que comme des intermédiaires de calcul). Il peut être toutefois différent pour le calcul des grandeurs thermodynamiques car la moyenne de l'énergie potentielle peut être calculée aux temps entiers (elle fait intervenir les positions), tandis que la moyenne de l'énergie cinétique est calculée aux temps demi-entiers (elle fait intervenir les vitesses). L’initialisation de la simulation en Dynamique Moléculaire se fait en fixant la position de départ des molécules et en leur donnant une vitesse initiale. Les équations de Newton impliquent ensuite une conservation de l’énergie au cours du temps. C'est donc naturellement dans l’ensemble micocanonique (c'est-à-dire à nombre de molécule N, à volume V et énergie E constants) que s’effectue une simulation en Dynamique Moléculaire.
d. Thermostats T et P
Toutefois, l’ensemble microcanonique n’est pas toujours représentatif des systèmes que l’on souhaite modéliser. On peut avoir besoin de travailler non plus à volume constant mais à pression constante. De plus expérimentalement, les systèmes sont plutôt en contact avec un thermostat et donc à température fixée plutôt qu’à énergie fixée. En effet, si l’on part d’un système qui n’est pas à l’équilibre et que l’on fait une simulation dans l’ensemble microcanonique, la stabilisation du système va s’accompagner d’un changement important de la température. Une façon simple de réguler la température du système est de multiplier, après chaque pas de calcul, les vitesses par un pas correctif T Tc, où T désigne la température thermodynamique que l’on souhaite imposer au système et Tc la température cinétique calculée à partir des vitesses. Tc (t ) = mi vi (t ) i =1 k B N f N ∑ 2 (II-79) Nf étant le nombre de degré de liberté, égal à (3N-3) pour un système de N particules avec une énergie totale constante.
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A Case of Unilateral Coccidioidal Chorioretinitis in a Patient with HIV-Associated Meningoencephalitis
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Christopher B. Toomey
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English
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Spoken
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Hindawi
Case Reports in Ophthalmological Medicine
Volume 2019, Article ID 1475628, 3 pages
https://doi.org/10.1155/2019/1475628 Hindawi
Case Reports in Ophthalmological Medicine
Volume 2019, Article ID 1475628, 3 pages
https://doi.org/10.1155/2019/1475628 1. Introduction Coccidioidomycosis is a disease caused most frequently by
Coccidioides immitis, a dimorphic fungus commonly found in
the lower Sonoran Desert ecozone of the Western hemisphere
[1]. Te disease, which typically manifests as a self-limiting
community-acquired pneumonia, was initially described in
the late 1800s and thought to be of protozoan etiology (and
hence the etymology of Coccidia) but was later identified as
a dimorphic fungus [2]. Endemic areas include Arizona, New
Mexico, West Texas, parts of Central America, Argentina,
Northwest Mexico, and the San Joaquin Valley in California
[1]. Populations at greater risk include pregnant women and
those with immunosuppressed conditions [3]. Symptomatic
patients usually present with 1–3 weeks afer exposure with
flu-like symptoms including night sweats, cough, myalgias,
and rash [4]. Christopher Toomey,1 Andrew Gross,2 Jeffrey Lee,1 and Doran B. Spencer
1
1Viterbi Department of Ophthalmology, Shiley Eye Institute, University of California at San Diego, San Diego, CA, USA
2McGovern Medical School, University of Texas at Houston Health Science Center, Houston, TX, USA Christopher Toomey,1 Andrew Gross,2 Jeffrey Lee,1 and Doran B. Spencer
1
1Viterbi Department of Ophthalmology, Shiley Eye Institute, University of California at San Diego, San Diego, CA, USA
2McGovern Medical School, University of Texas at Houston Health Science Center, Houston, TX, USA Correspondence should be addressed to Doran B. Spencer; dbspencer@ucsd.edu Received 8 July 2019; Accepted 29 August 2019; Published 7 October 2019 Received 8 July 2019; Accepted 29 August 2019; Published 7 October 2019 Academic Editor: Claudio Campa Copyright © 2019 Christopher Toomey et al. Tis is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Intraocular coccidioidomycosis is a rare condition, with the most commonly reported presentation being an idiopathic iritis in
patients who live in or have traveled thorough endemic areas. A paucity of reports exists describing the chorioretinal manifestations of
coccidioidomycosis. Here we report a case of unilateral coccidioidal chorioretinitis and meningoencephalitis in an AIDS patient that
led to near complete unilateral loss of vision. A 48-year-old Hispanic female with poorly controlled HIV/AIDS in southern California
presented with a three-week history of headache, nausea, vomiting, right eye blurry vision, and a one-day history of subjective
fever. Examination of the right eye revealed vitritis and several large chorioretinal lesions scattered throughout the periphery and
macula with optic disc pallor. Serum coccidioidomycoses complement fixation (CF) was positive (titers of 1 : 256). Neuroimaging
revealed a new area of enhancement in the lef anterior frontal lobe consistent with meningoencephalitis. Te patient was treated
with intravenous fluconazole and intravitreal voriconazole with resolution of systemic symptoms and vitritis but persistence of
unilateral, severe chorioretinal scarring and vision loss. In conclusion, in spite of the rarity of intraocular coccidioidomycosis, one
must carry a degree of suspicion for this vision- and life-threatening condition as a potential etiology of chorioretinitis in individuals
with pertinent risk factors. endemic areas [5, 6]. A paucity of reports exists describing the
chorioretinal manifestations of coccidioidomycosis. We report
an unusual, unilateral case of coccidioidal chorioretinitis with
panuveitis and meningoencephalitis in a patient with AIDS
that led to near complete, unilateral loss of vision. 2. Case Presentation A 48-year-old Hispanic female presented to a HIV clinic in
southern California with a three-week history of headache,
right eye blurry vision, dizziness, myalgia, nausea, and vom-
iting, and a one-day history of subjective fever. Te past med-
ical history was significant for human immunodeficiency virus
(HIV) for 13 years (CD4 103, viral load 877 two weeks prior)
with chronic associated cytomegalovirus (CMV) viremia,
mycobacterium avium-intracellulare infection and toxoplas-
mosis encephalitis. Te patient’s medications included abaca-
vir, dolutegravir, and lamivudine, as well as azithromycin,
ethambutol, pyrimethamine, sulfadiazine, and valganciclovir Uveitis secondary to coccidioidomycosis is rare, and the
few reports of it most frequently describe an idiopathic, bilat-
eral iritis in patients who live or have traveled thorough Case Reports in Ophthalmological Medicine 2 Figure 1: Fundus photograph with a yellow arrow marking an old, atrophic choroidal scar and blue lines delineating multiple white/off-
yellow exudates around the macular and extending into the peripheral retina. Pallor of the optic disc is notable as well as minor vitreous haze. Figure 2: Axial T1 with contrast (lef) and axial T2 FLAIR (right) magnetic resonance imaging (MRIs) demonstrating a peripherally enhancing
5 mm lesion in the lef frontal lobe surrounded by moderate confluent vasogenic edema, marked by red arrows. Figure 1: Fundus photograph with a yellow arrow marking an old, atrophic choroidal scar and blue lines delineating multiple white/off-
yellow exudates around the macular and extending into the peripheral retina. Pallor of the optic disc is notable as well as minor vitreous haze. Figure 1: Fundus photograph with a yellow arrow marking an old, atrophic choroidal scar and blue lines delineating multiple white/off-
yellow exudates around the macular and extending into the peripheral retina. Pallor of the optic disc is notable as well as minor vitreous haze. Figure 2: Axial T1 with contrast (lef) and axial T2 FLAIR (right) magnetic resonance imaging (MRIs) demonstrating a peripherally enhancing
5 mm lesion in the lef frontal lobe surrounded by moderate confluent vasogenic edema, marked by red arrows. Figure 2: Axial T1 with contrast (lef) and axial T2 FLAIR (right) magnetic resonance imaging (MRIs) demonstrating a peripherally enhancing
5 mm lesion in the lef frontal lobe surrounded by moderate confluent vasogenic edema, marked by red arrows. for opportunistic infections. Her temperature was 98.1°F, she
had no abnormal vital signs, no recent weight change, no neu-
rological symptoms and a normal systemic physical exam. were negative. References [1] F. S Fisher, M. W Bultman, S. M Johnson, D. Pappagianis, and
E. Zaborsky, “Coccidioides niches and habitat parameters in the
Southwestern United States: a matter of scale,” Annals of the
New York Academy of Sciences, vol. 1111, no. 1, pp. 47–72, 2007. [2] W. Ophuls, “Further observations on a pathogenic mould
formerly described as a protozoon (Coccidioides immitis,
Coccidioides pyogenes),” Journal of Experimental Medicine,
vol. 6, no. 4–6, pp. 443–485, 1905. [3] C. D. Odio, B. E. Marciano, J. N. Galgiani, and S. M. Holland,
“Risk factors for disseminated coccidioidomycosis, United
States,” Emerging Infectious Diseases, vol. 23, no. 2, 2017. [4] J. N. Galgiani, N. M. Ampel, J. E. Blair et al., “Coccidioidomycosis,”
Clinical Infectious Diseases, vol. 41, no. 9, pp. 1217–1223, 2005. [5] H. T. Rodenbiker and J. P. Ganley, “Ocular coccidioidomycosis,”
Survey of Ophthalmology, vol. 24, no. 5, pp. 263–290, 1980. Te patient in this report presented with severe monocular
vision loss with significant chorioretinal lesions and optic atro-
phy along with significant vitritis and a positive serum coc-
cidioidomycoses CF. CSF coccidioidomycoses CF was
negative, however, CSF testing has a low sensitivity in patients
with coccidioidomycoses meningitis [13]. Our case report
highlights the severity of coccidioidal chorioretinitis that can
be associated with meningoencephalitis in a patient with HIV/
AIDS, including in a unilateral fashion. Given the fundus
appearance of old and new lesions, our clinical impression is
that the chorioretinal involvement was on-going for many
months and the patient presented for ophthalmologic evalu-
ation once her central vision was affected. Tis supports the
notion that ocular involvement of coccidioidomycosis may be
an insidious process and highlights the essential role of routine
ophthalmologic evaluation in AIDS patients in aiding in the
diagnosis of systemic disease. [6] R. S. Moorthy, N. A. Rao, Y. Sidikaro, and R. Y. Foos,
“Coccidioidomycosis Iridocyclitis,” Ophthalmology, vol. 101,
no. 12, pp. 1923–1928, 1994. [7] R. D. Schrier, W. R. Freeman, C. A. Wiley, and J. A. McCutchan,
“Immune predispositions for cytomegalovirus retinitis in
AIDS. Te HNRC group,” Journal of Clinical Investigation,
vol. 95, no. 4, pp. 1741–1746, 1995. [8] K. A. Zakka, R. Y. Foos, and W J. Brown, “Intraocular
coccidioidomycosis,” Survey of Ophthalmology, vol. 22, no. 5,
pp. 313–321, 1978. [9] D. V. Vasconcelos-Santos, J. I. Lim, and N. A. Rao, “Chronic
coccidioidomycosis endophthalmitis without concomitant
systemic involvement: a clinicopathological case report,”
Ophthalmology, vol. 117, no. 9, pp. 1839–1842, 2010. Conflicts of Interest vitreous sampling. During her hospital course her systemic
symptoms resolved. Follow-up examinations with serial
fundus examinations showed progressive improvement in
posterior vitritis, however, her visual acuity and retinal lesions
remained stable throughout her hospital course. On follow-up
examination at 3 and 5 months afer discharge, her vision had
decreased to light perception with no evidence of active lesions
or vitritis and stable chorioretinal lesions. No conflicting relationships exist for any author. Authors’ Contributions CBT, AG, JL, and DBS were responsible for data collection,
analysis, and writing/preparation of the manuscript. Tis work was supported by the NIH NEI P30 P30EY0225789
Vision Research Core. Tis case report describes the clinical manifestations of a
patient with an unusual presentation of unilateral coccidioidal
panuveitis with chorioretinitis; to our knowledge, this is
unique in the literature. Few clinical reports exist describing
the intraocular clinical manifestations of coccidioidomycosis
[5, 6, 8–11]. Rodenbiker and Ganley reviewed a series of cases
of ocular coccidioidomycoses with only 10 reported to have a
posterior uveitis (retinitis, choroiditis, or chorioretinitis). Te
authors noted the anterior segment was ofen uninvolved and
fundus findings included a range of lesions with yellow-white
exudate ranging from <1 to 3 disc diameters [5]. Prevalence
of asymptomatic chorioretinal scars in patients with coccidi-
oidomycosis have been reported in 5 of 54 subjections (9.2%),
suggesting the prevalence of coccidioidal chorioretinitis is
higher than isolated reports suggest [12]. However, it is diffi-
cult to make assumptions on the prevalence of coccidioidal
chorioretinitis in this manner given the difficulty of linking
the chorioretinal scarring to the coccidioidomycosis. References [10] E. T. Cunningham, “Intraocular coccidioidomycosis diagnosed
by skin biopsy,” Archives of Ophthalmology, vol. 116, no. 5,
pp. 674–677, 1998. In spite of the rarity of intraocular coccidioidomycosis,
recognizing the clinical appearance of this potentially vision
and life-threatening condition is essential in individuals with
pertinent risk factors including pregnancy and immunosup-
pression, along with a history of travel or living in endemic
areas [1, 3]. [11] R. A. Shields, P. H. Tang, Z. M. Bodnar, S. J. Smith, and
A. R. Silva, “Optical coherence tomography angiography
highlights chorioretinal lesions in ocular coccidioidomycosis,”
Ophthalmic Surgery, Lasers and Imaging Retina, vol. 50, no. 3,
pp. e71–e73, 2019. [12] H. T. Rodenbiker, J. P. Ganley, J. N. Galgiani, and S. G. Axline, “Prevalence of chorioretinal scars associated with
coccidioidomycosis,” Archives of Ophthalmology, vol. 99,
no. 1, pp. 71–75, 1981. 2. Case Presentation Gram stain showed polymorphonucleated
white blood cells without organisms and cultures showed no
growth. Te encephalitis panel polymerase chain reaction was
performed on the vitreous tap sample and was negative for
Enterovirus, HSV1/2, Herpesvirus 6, Parechovirus, Varicella
Zoster and Cryptococcus neoformans/gatti, E-coli K1,
Haemophilus influenza, Listeria monocytogenes, Neisseria men-
ingitides, Strep. agalactiae, Strep. pneumoniae. Te vitreous
PCR test was positive for Cytomegalovirus. CT head showed
a new area of enhancement in the lef anterior frontal lobe
(Figure 2). Lumbar puncture (LP) with CSF gram stains, fun-
gal cultures, and coccidioides CF were negative. Ophthalmic examination revealed visual acuity of hand
motion at 1 foot in the right eye (OD) and 20/20 in the lef eye
(OS), pupil constriction of 3 > 3 mm OD and 3 > 2 mm OS, and
a positive right-sided relative afferent pupillary defect (RAPD). Intraocular pressures were within normal limits and extraoc-
ular movements were full in both eyes. Confrontational visual
fields and color plates were unable to be obtained OD and
within normal limits OS. Anterior segment examination was
within normal limits on bedside exam. Dilated fundus exam
OD revealed 1+ vitreous cell/haze, several large chorioretinal
lesions scattered throughout the periphery and involving the
macula with 2+ optic disc pallor. OS was within normal limits
(Figure 1). g
g
Based on the symptoms, positive serum coccidioidomycoses
complement fixation and fundus appearance the patient was
diagnosed with coccidioidal chorioretinitis with panuveitis. Te vitreous PCR positive test was attributed to her long-
standing CMV viremia secondary to sample contamination
from the vitreous cells and trauma of the intravitreal biopsy
and not deemed consistent with CMV retinitis given the
absence of suspicious lesions. Additionally, her CD4 count of
103 places her outside of the highest risk strata of CMV
retinitis, which occurs most commonly at a CD4 count of <50
[7]. Te patient was treated with intravenous fluconazole and
intravitreal voriconazole was preemptively given during the Due to the immunosuppressive state on presentation, as
well as systemic symptoms, an extensive infectious work-up
was performed as an inpatient with the Infectious Disease
service and a vitreous tap was performed of the right eye. Serum coccidioidomycoses complement fixation (CF) was
found to be positive (titers of 1 : 256). Serum syphilis enzyme
immunoantibody, rapid plasma reagin, Borrelia burgdorferi
antibody, CMV IgG/IgM, Varicella IgG, HSV type 1/2 IgM Case Reports in Ophthalmological Medicine 3 3. Discussion Tis work was supported by the NIH NEI P30 P30EY0225789
Vision Research Core. Tis work was supported by the NIH NEI P30 P30EY0225789
Vision Research Core. Ethical Approval Te authors have no ethical conflicts to disclose. Tis research
has been approved by the UCSD committee on human
research. [13] G. Mathisen, A. Shelub, J. Truong, and C. Wigen, “Coccidioidal
meningitis,” Medicine, vol. 89, no. 5, pp. 251–284, 2010.
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A mobilidade na aprendizagem: uma nova dimensão para a aprendizagem de língua estrangeira mediada por telefone celular
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Lucía Silveira Alda
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Portuguese
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Spoken
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http://periodicos.letras.ufmg.br/index.php/textolivre
Ano: 2014 – Volume: 7 – Número: 1 http://periodicos.letras.ufmg.br/index.php/textolivre
Ano: 2014 – Volume: 7 – Número: 1 A MOBILIDADE NA APRENDIZAGEM: UMA NOVA DIMENSÃO PARA A
APRENDIZAGEM DE LÍNGUA ESTRANGEIRA MEDIADA POR TELEFONE CELULAR Lucía Silveira Alda/Universidade Católica de Pelotas RESUMO: As tecnologias digitais estão tornando-se cada vez mais presentes nos ambientes
sociais, modificando a maneira como nos comportamos e, consequentemente, como aprendemos. À
medida que o ser humano evolui, criam-se novos hábitos e, com estes, necessitamos enfrentar novos
desafios. A integração das novas tecnologias ao nosso cotidiano e o acesso facilitado à informação
provocam mudanças tecnológicas que influenciam novos paradigmas educacionais emergentes,
como a aprendizagem móvel. Dessa forma, este trabalho objetiva esclarecer o que é e como
funciona esse modelo de aprendizagem. Para isso, fez-se uma revisão da literatura da área,
revisitando diversos conceitos e verificando a sua evolução, a fim de refletir sobre possíveis
caminhos, destacando as principais potencialidades ao se trabalhar com a aprendizagem móvel. São
ainda apontadas sugestões de aplicação da aprendizagem móvel utilizando um dos principais
dispositivos tecnológicos atuais, o telefone celular, como ferramenta de aprendizagem de línguas. ALAVRAS-CHAVE: Aprendizagem móvel. Aprendizagem de línguas. Telefone celular PALAVRAS-CHAVE: Aprendizagem móvel. Aprendizagem de línguas. Telefone celular. ABSTRACT: Digital technologies are becoming increasingly present in social environments,
changing the way we behave and consequently, the way we learn. As humans evolve, new habits are
created and, with these, we need to face new challenges. The integration of new technologies to our
everyday life and the easy access to information cause technological changes that influence new
emerging educational paradigms, such as mobile learning. Thus, this study aims to clarify what is
this learning model and how it works. For this, the literature of this area was reviewed, revisiting
many concepts and verifying its evolution to reflect on possible ways, highlighting the main
capabilities when working with mobile learning. Still, some suggestions are pointed for
implementing mobile learning using one of the main current technological devices, the mobile
phone, as a language learning tool. KEYWORDS: Mobile learning. Language Learning. Mobile Phone. KEYWORDS: Mobile learning. Language Learning. Mobile Phone. INTRODUÇÃO A necessidade de comunicação é inerente ao ser humano. A troca de informações, o
registro dos fatos, a expressão das ideias e das emoções contribuíram, desde os tempos mais
remotos, para a evolução dos formatos e surgimento de inúmeras plataformas relacionadas à
comunicação. A partir do século 20, os avanços tecnológicos popularizaram o acesso à informação,
contribuindo para a disseminação das tecnologias de informação e comunicação (TIC). No campo
da aprendizagem de línguas estrangeiras, a tecnologia sempre buscou facilitar a prática humana,
expor o aluno a um contexto estrangeiro e transpor as barreiras geográficas, além de fornecer
suporte à aprendizagem. Dessa forma, não podemos ignorar esses avanços tecnológicos, já que,
quando explorados corretamente, podem auxiliar bastante no processo de aprendizagem de uma 98 http://periodicos.letras.ufmg.br/index.php/textolivre
Ano: 2014 – Volume: 7 – Número: 1 http://periodicos.letras.ufmg.br/index.php/textolivre
Ano: 2014 – Volume: 7 – Número: 1 língua estrangeira. Além disso, as tecnologias também tentam acompanhar as necessidades humanas: hoje,
o ser humano é móvel e requer ferramentas capazes de acompanhar o seu cotidiano. Esse fator
influenciou na evolução tecnológica, que possibilitou desenvolver ferramentas mais leves, menores
e muito mais eficientes, além de cada vez mais acessíveis. A tecnologia móvel não é exceção na
vida da maioria dos alunos e, a todo o momento, seja dentro ou fora do ambiente escolar, o acesso a
ela é cada vez maior e mais comum. Ferramentas como o telefone celular contribuem para que
estejamos cada vez mais conectados, com a possibilidade de comunicação com outras pessoas em
qualquer momento, onde quer que estejamos; agora é natural, principalmente para as novas
gerações, acessar informação, tirar fotografias, compartilhar momentos com amigos, colegas ou
com o resto do mundo. Os avanços da tecnologia móvel e sem fio e dos dispositivos computacionais portáteis,
aliados à interação entre pessoas distantes geograficamente e fisicamente, proporcionam novos
conceitos e processos de aprendizagem. A aprendizagem móvel surge como uma nova possibilidade
educacional, potencializada pela massificação do uso de tecnologias móveis, podendo eliminar
algumas limitações de aprendizagem que existem dentro do ambiente escolar, proporcionando o
acesso a materiais de ensino independentemente do local e do tempo. Apesar de tudo, é evidente
também que, ao lado de tantas potencialidades, torna-se necessário refletir sobre prováveis desafios
a serem enfrentados por professores de língua estrangeira. Logo, devemos discutir os novos
caminhos que se abrem e refletir sobre novas propostas de ensino. INTRODUÇÃO Assim, este artigo objetiva
apresentar os principais conceitos sobre aprendizagem móvel, fazendo uma revisão da literatura da
área, e expor como esta se apresenta em contexto de aprendizagem de línguas e suas possibilidades
de uso através de dispositivos móveis, como o telefone celular, a fim de discutir suas vantagens,
limitações e alcances metodológicos. 1 CONCEITOS EM APRENDIZAGEM MÓVEL A noção de aprendizagem móvel, fundamentalmente denominada m-learning ou mobile
learning, ainda está sendo instituída. As pesquisas sobre esse paradigma de aprendizagem ainda são
emergentes e, portanto, há uma grande diversidade de conceitos entre os pesquisadores da área; no
entanto, é possível sumarizar suas semelhanças. O conceito de aprendizagem móvel surge em um
contexto onde se procura explorar de que maneira as tecnologias móveis podem auxiliar na
aprendizagem e explicar a relação complexa entre os desenvolvimentos tecnológicos e seus
potenciais para educação e aprendizagem. A aprendizagem móvel é um processo que se dá através
de múltiplos contextos entre pessoas e tecnologias móveis, interativas e pessoais (SHARPLES;
TAYLOR; VAVOULA, 2007). Para Kukulska-Hulme e Traxler (2005, p. 42), a aprendizagem móvel pode ser espontânea, portátil, pessoal, situada; pode ser informal, conveniente, onipresente e
perturbadora. Nos aproxima mais de uma aprendizagem ‘a qualquer hora, em qualquer
lugar’, mas ainda é muito cedo para prever como nosso entendimento de ensino e
aprendizagem irá evoluir como consequência. Por sua vez, Chabra e Figueiredo (2002) percebem a aprendizagem móvel como uma
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Ano: 2014 – Volume: 7 – Número: 1 qualquer aparelho, enfatizando o grande fator de destaque deste paradigma educacional: a
mobilidade. Isso acontece devido a esses processos de aprendizagem ocorrerem, necessariamente,
apoiados pelo uso de tecnologias portáteis, móveis e sem fio (SCHLEMMER; SACCOL;
BARBOSA; REINHARD, 2007; TRAXLER, 2009). Dessa forma, a aprendizagem móvel pode ser
inferida como qualquer aparelho, enfatizando o grande fator de destaque deste paradigma educacional: a
mobilidade. Isso acontece devido a esses processos de aprendizagem ocorrerem, necessariamente,
apoiados pelo uso de tecnologias portáteis, móveis e sem fio (SCHLEMMER; SACCOL;
BARBOSA; REINHARD, 2007; TRAXLER, 2009). Dessa forma, a aprendizagem móvel pode ser
inferida como a intersecção entre computação móvel e e-learning: recursos acessíveis onde quer que você
esteja, forte capacidade de pesquisa, rica interação, forte apoio para uma aprendizagem
eficaz e baseada na performance. Aprendizagem independente da localização no tempo ou
espaço (QUINN, 2000, p. 1). Os resultados de um estudo desenvolvido por Attewell (2005) indicam que a
aprendizagem móvel permite uma aprendizagem personalizada, em qualquer hora e lugar. Esta pode
ser utilizada para adicionar variedade às aulas e remover um pouco da formalidade, proporcionando
maior ludicidade. 2 MOBILIDADE: UMA NOVA DIMENSÃO DE APRENDIZAGEM A capacidade técnica dos novos dispositivos móveis e a qualidade de ser móvel
adicionam uma nova dimensão à aprendizagem e prática de línguas estrangeiras. Klopfer, Squire e
Jenkins (2002) identificaram cinco propriedades de affordancei únicas para a educação e todas
plenamente compreendidas pela aprendizagem móvel: a (1) portabilidade – o tamanho e o peso, que
permitem carregar e mover o dispositivo para todo lado; a (2) interatividade social – troca de dados
e colaboração com outros alunos; a (3) sensibilidade ao contexto – interação de acordo com o
ambiente, local e hora, incluindo dados reais ou simulados; a (4) conectividade – ambiente em rede
e compartilhamento de informações entre dispositivos e alunos; e a (5) individualidade – atividades
personalizadas. Ribeiro, Ferreira e Carneiro (2011) apresentam pressupostos nos quais assentam
vantagens ao se adotar a aprendizagem móvel. Primeiramente, a possibilidade de interação entre
professor-aluno-aluno. Logo, a portabilidade e a colaboração (a primeira permite a mobilidade, e o
segundo o trabalho em conjunto de uma tarefa, mesmo estando os sujeitos em locais distantes um
do outro). Ainda, a promoção do compromisso dos alunos, dada a grande aceitação de dispositivos
móveis, aumentando assim a motivação. Por fim, a melhoria da autonomia e flexibilidade. Outra
vantagem da aprendizagem móvel é o acesso à informação através dos dispositivos móveis, como
também a comodidade e a rapidez, que permitem uma maior interação, tanto entre membros de um
grupo de estudantes, como de forma direta com o professor, em tempo real. A tecnologia está em constante evolução e cada vez mais adaptada ao nosso cotidiano e
às nossas necessidades. Os avanços tecnológicos possibilitam, hoje, ferramentas mais leves,
menores e muito mais eficientes, além de cada vez mais acessíveis, e a mobilidade que esses novos
dispositivos proporcionam propõe uma nova dimensão de aprendizagem na área de línguas
estrangeiras: Hoje, a tecnologia é dinâmica, móvel, miniaturizada em circuitos integrados. O que cabia
na sala então, hoje pode ser levado no bolso da camisa, incluindo não apenas áudio, mas
também texto escrito e vídeo, a um custo tão baixo e com um benefício tão alto que ficou
bem mais difícil rejeitá-la (LEFFA, 2009, p. 15). O avanço acelerado das novas tecnologias no nosso cotidiano, aliado à mobilidade e
facilidade de acesso, faz com que muitas ferramentas sejam cada vez mais naturais, tornando-se
“invisíveis”; as ferramentas móveis estão ficando tão incorporadas às nossas vidas, que é impossível
ignorá-las. Leffa (2009, p. 1 CONCEITOS EM APRENDIZAGEM MÓVEL Também mostra-se eficiente para auxiliar na alfabetização e ensino de línguas,
ajudando alunos e professores a reconhecer e desenvolver as competências básicas. Além disso, a
aprendizagem móvel facilita experiências individuais e colaborativas e pode ajudar a aumentar a
autoconfiança e autoestima do aluno, permitindo uma experiência de aprendizagem personalizada. A aprendizagem móvel pode ajudar, ainda, a combater a resistência existente ao uso das TIC,
fornecendo uma ponte entre o ensino de línguas e as tecnologias móveis. O conceito de ubiquidade também é associado ao conceito de m-learning, em que u-
learning, ou aprendizagem ubíqua, apresenta-se como um desenvolvimento de teorias que procuram
reunir os conceitos de e-learning e m-learning. De acordo com Clarey (2007), a aprendizagem ubíqua (u-learning) utiliza aparelhos inteligentes para proporcionar às
pessoas a informação correta no tempo e de maneira certa – é a qualquer hora, em qualquer
lugar (literalmente), de qualquer jeito. [...] é a convergência colaborativa e informal de e-
learning e m-learning. É uma aprendizagem “usável” com uma característica social que
embute a aprendizagem no nosso trabalho e/ou vida. Segundo Ribeiro, Ferreira e Carneiro (2011), a aprendizagem móvel pode ainda ser
compreendida como uma evolução de e-learning: e-learning é estático; nele, as atividades são
realizadas em ambientes de aprendizagem acessíveis por redes de computadores ou espaços
computadorizados fechados e privados, e a internet é restrita; em contrapartida, a aprendizagem
móvel acontece em qualquer hora e lugar, utilizando dispositivos móveis e sem fio para promover a
comunicação e a interação, as mensagens são instantâneas e não há limites geográficos, a internet é
acessível, sem fio ou 3G. E-learning baseia sua intervenção em documentos (textos e gráficos) e aulas virtuais,
enquanto que no m-learning há maior comunicação verbal e as aulas são em contexto real;
[...] enquanto que com o e-learning a comunicação não é instantânea (e-mail e websites), no
caso do m-learning a comunicação é imediata quer por e-mail quer por SMS. No e-
learning, é necessário calendarizar tarefas e no caso do m-learning, o processo é
espontâneo (RIBEIRO; FERREIRA; CARNEIRO, 2011, p. 10). A aprendizagem móvel busca propiciar a aprendizagem independentemente do tempo
ou espaço, utilizando ferramentas portáteis que possibilitem ao aluno fazer escolhas quanto à sua 100 http://periodicos.letras.ufmg.br/index.php/textolivre
Ano: 2014 – Volume: 7 – Número: 1 maneira de aprender, caracterizando-se como uma aprendizagem particularmente flexível e
adaptável e, por isso, oferece diversas possibilidades de aplicação. 2 MOBILIDADE: UMA NOVA DIMENSÃO DE APRENDIZAGEM 13) afirma que “o destino de qualquer instrumento, tecnológico ou
psicológico, é a invisibilidade”. Logo, ao dominar um instrumento de aprendizagem, o interesse
volta-se ao objetivo que se pretende atingir através daquela ferramenta, e não à ferramenta em si,
tornando-a invisível. Reforçando ainda essa ideia, Weiser (1991) declara que esse
“desaparecimento” da ferramenta, a invisibilidade, é uma consequência fundamental da psicologia
humana, e não da tecnologia. Sempre que alguém aprende algo suficientemente bem, deixa de ter
consciência disso; isto é, a ferramenta torna-se tão natural na vida dos sujeitos, que é utilizada quase
sem pensar. Essas noções de invisibilidade equiparam-se ao conceito de naturalização de Bax 101 http://periodicos.letras.ufmg.br/index.php/textolivre
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Ano: 2014 – Volume: 7 – Número: 1 (2003), que afirma que o uso desses dispositivos tecnológicos será tão natural quanto as demais
tecnologias existentes no ambiente educacional, como o quadro e a caneta, por exemplo. Toda essa mudança tecnológica influencia as novas gerações que surgem, e cada vez faz
mais sentido aproximar as tecnologias móveis dos ambientes de aprendizagem. Elas estão
transformando os hábitos das pessoas, a maneira como trabalham, ensinam e aprendem. Segundo
Prensky (2001), os nossos alunos mudaram radicalmente. Os alunos de hoje não são mais as
pessoas para as quais o nosso sistema educacional foi projetado para ensinar; alguns professores
supõem que os alunos são os mesmos de sempre, e que os mesmos métodos que funcionaram para
os professores quando estes eram alunos irão funcionar para os seus alunos hoje. Porém, essa
suposição não é mais válida, os alunos hoje são diferentes, e, por isso, a era tecnológica necessita de
um sistema educacional reformulado voltado para esses novos alunos, os “nativos digitaisii”: Eles passaram a vida inteira cercados por e utilizando computadores, videogames,
reprodutores de música digital, câmeras de vídeo, celulares, e todos os outros brinquedos e
ferramentas da era digital. [...] Jogos de computador, e-mail, internet, celulares e mensagens
instantâneas são partes integrais de suas vidas (PRENSKY, 2001, p. 1). Essa nova geração está acostumada a dividir a sua atenção entre diferentes tarefas ao
mesmo tempo, utilizando diferentes tipos de tecnologias e inserida em diferentes tipos de contexto;
o conteúdo acessado e produzido pelos nativos digitais não se limita a textos, abrange também
imagens, sons, vídeos e multimídias. Saccol, Schlemmer e Barbosa (2011, p. 2 MOBILIDADE: UMA NOVA DIMENSÃO DE APRENDIZAGEM 21) complementam
que “para essa nova geração, a educação tradicional, centrada no professor, desenvolvida de forma
linear, fundamentalmente baseada em texto e excessivamente expositiva, não faz sentido. A nova
geração está acostumada a agir em vez de passivamente assistir”. É evidente também que, ao lado de tantas potencialidades, torna-se necessário refletir
sobre prováveis desafios que podem vir a ser enfrentados por professores de língua estrangeira,
entre os quais destacam-se a necessidade de letramento digital e a resistência ao uso de novas
tecnologias. Não basta apenas ter a ferramenta, pois a ferramenta por si só não é suficiente. É
necessário repensar a questão metodológica: Um dos pontos mais frágeis identificados por diferentes pesquisadores, no que se refere a
essas modalidades educacionais, é a questão didático-pedagógica. Não basta ter acesso a
novas tecnologias que possam ser utilizadas de forma combinada; é preciso, sobretudo,
saber como utilizá-las para propiciar a aprendizagem dos sujeitos (SACCOL;
SCHLEMMER; BARBOSA, 2011, p. 2). Existem inúmeras possibilidades de aprendizagem utilizando ferramentas móveis,
porém os educadores devem repensar e redesenhar as atividades de forma que sejam eficientes para
um determinado sujeito nesse contexto tecnológico. Segundo Ribeiro, Ferreira e Carneiro (2011, p. 4), “as tecnologias móveis aliadas a uma metodologia adequada ao meio podem contribuir para uma
prática pedagógica focada no desenvolvimento de competências numa perspectiva de diálogo e
cooperação”. 3 (ALGUMAS) POSSIBILIDADES DO TELEFONE CELULAR A partir dessa incontestável disseminação de inúmeras ferramentas tecnológicas móveis 102 http://periodicos.letras.ufmg.br/index.php/textolivre
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Ano: 2014 – Volume: 7 – Número: 1 e da sua disponibilidade no mercado, o telefone celular destaca-se pelas suas taxas de difusão,
popularização e acesso, podendo ser utilizado para a aprendizagem e prática de idiomas em
qualquer situação, lugar e hora. Como a grande maioria das tecnologias, a tecnologia de telefonia
celular também evoluiu rapidamente. Além da evolução dos padrões e tecnologias móveis, foram
sendo incorporados aos aparelhos móveis inúmeros recursos, como câmeras, mecanismos de
localização, gravadores de voz, editores de texto e diversos recursos computacionais. As estatísticas
acerca dos telefones celulares ainda corroboram a potencialidade do aparelho, demonstrando que
este tem (1) a acessibilidade praticamente universal, na medida em que atualmente a maioria da
população possui pelo menos um celular; (2) o suporte a inúmeros recursos do próprio aparelho,
como áudio, vídeo e acesso à internet; (3) a possibilidade de trabalhar as quatro habilidades da
língua (escuta, fala, leitura e escrita); e (4) a possibilidade de acesso em qualquer hora e lugar. Segundo uma pesquisa sobre os itens mais importantes do dia a dia, realizada pelo
IBOPE Mídia, o telefone celular aparece em segundo lugar no ranking de prioridades, perdendo
apenas para o aparelho de televisão. No entanto, fica à frente do computador com internet e do
rádio. Hoje, existem sete bilhões de pessoas no planeta e o número de celulares demonstra que as
taxas de difusão do telefone celular superam as da maioria das tecnologias anteriores, incluindo os
computadores pessoais. No Brasil, dados da Anatel indicam que o mês de abril de 2014 terminou
com 273,6 milhões de celulares e uma densidade de 135,2 celulares a cada 100 habitantes
(TELECO, 2014). Como mencionado anteriormente, uma das principais características do telefone celular
é a sua mobilidade e a sua indiscutível popularidade, o que lhe confere uma grande vantagem diante
de outras ferramentas como plataforma do futuro. Embora exista no mundo uma grande quantidade
de telefones celulares, estes ainda não adquiriram notabilidade no campo da educação. Essa
ferramenta permite ao aluno trabalhar com as quatro habilidades básicas de uma língua estrangeira
através das diversas funcionalidades disponíveis nos aparelhos, até mesmo nos mais simples. 3 (ALGUMAS) POSSIBILIDADES DO TELEFONE CELULAR Podemos destacar, por exemplo, a utilização de short message service (doravante SMS), um dos
recursos mais relevantes dos aparelhos celulares – nos Estados Unidos, apenas em um dia, são
recebidos duzentos trilhões de SMS, o que é mais do que um ano inteiro de cartas recebidas. Esse
serviço existe em todos os aparelhos celulares e possibilita trocar (enviar e receber) mensagens
escritas com diversas pessoas, sendo adequado para trabalhar as habilidades de leitura e escrita. Outro recurso possível existente em todos os celulares é o de voz, pois a função
primeira dessa ferramenta é a capacidade de efetuar ligações, o que exercita tanto a habilidade de
fala quanto a de escuta. Segundo Prensky (2005), até os celulares mais simples têm chips mais
poderosos e complexos do que o computador a bordo da nave espacial que pousou na lua em 1969. Em celulares mais avançados, existem os recursos de gravação de voz, execução de gravações e
músicas, entre outros. Além desses recursos, também existem outras funcionalidades, como acesso à
internet – e consequentemente, acesso a redes sociais, microblogs e inúmeras possibilidades –,
consulta a dicionários, troca de e-mails, criação e visualização de vídeos, compartilhamento de
vídeos e imagens, recursos de fotografia, leitura de arquivos, sistemas de localização por GPS e
acesso a mapas. Enfim, com apenas alguns desses recursos, é possível pensar em muitas maneiras
de aprender e ensinar uma língua estrangeira. Embora os aparelhos celulares sejam ferramentas relativamente fáceis – de fácil acesso,
comuns e populares, que não exigem grandes conhecimentos para poderem ser utilizadas, na
medida em que já estão incorporadas ao nosso cotidiano – são praticamente inutilizados no campo
educacional. Além de a maioria dos institutos escolares proibirem o uso de telefones celulares nos
seus ambientes, alguns professores acreditam que essa ferramenta atrapalha e distrai os alunos, que 103 http://periodicos.letras.ufmg.br/index.php/textolivre
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Ano: 2014 – Volume: 7 – Número: 1 estão mais interessados no aparelho telefônico do que nas próprias aulas. Mas, se existe o interesse
dos alunos, por que não explorá-lo? Sabemos que não é tão simples assim, já que tudo o que é novo
tende a despertar tanto interesse quanto medo. 3 (ALGUMAS) POSSIBILIDADES DO TELEFONE CELULAR Por isso, nossa responsabilidade como pesquisadores
e educadores é analisar os novos paradigmas educacionais emergentes, como a aprendizagem
móvel, e debater acerca de prováveis desafios a serem enfrentados, como, por exemplo, maneiras de
inserir o telefone celular de modo producente e efetivo em ambientes escolares, onde ele está
explicitamente proibido. Além disso, é necessário considerar que, assim como apresenta grandes
vantagens, a aprendizagem móvel também pode apresentar algumas vulnerabilidades em seu uso,
devido à grande variedade de plataformas, dispositivos e tecnologias, o que torna difícil unificar e
padronizar o processo. A tecnologia avança e se atualiza muito rápido, dificultando o
acompanhamento e prolongando o tempo necessário de pesquisa. Ainda, é fundamental estar consciente de que a ferramenta por si só não é suficiente. O
que faz o dispositivo eficiente na aprendizagem é o planejamento das atividades e a sua correta
aplicação do ponto de vista didático-metodológico. Existem inúmeras possibilidades de
aprendizagem com a utilização de telefones móveis, porém os educadores devem repensar e
redesenhar as atividades de forma que sejam eficientes para um determinado sujeito nesse contexto
tecnológico. CONSIDERAÇÕES FINAIS Como alunos, devemos utilizar as ferramentas em benefício da aprendizagem. Devemos
nos apropriar de novas tecnologias que visem facilitar o acesso à informação e ao
compartilhamento, que permitam a interatividade e que visem também à autonomia. Como
professores, devemos estar em contínua formação, acompanhando o desenvolvimento das
tecnologias que surgem, e também utilizá-las para nosso benefício. Devemos nos conscientizar e
conscientizar os nossos alunos das possíveis vantagens e desvantagens de utilizar certas
ferramentas. Não devemos limitar a aprendizagem ao ambiente escolar. A aprendizagem é o aqui e o
agora, independentemente de localização ou tempo. Devemos incitar a curiosidade pelo
conhecimento e a troca de informações. Hoje, a evolução das tecnologias e da sociedade exige
professores que direcionem o conhecimento com o aluno, de uma forma coletiva, e não apenas
unilateral. A aprendizagem móvel instiga exatamente isso. Com ela, podemos explorar o acesso à
informação com a comodidade que a mobilidade proporciona e a rapidez dos recursos tecnológicos,
que permitem uma maior interação. Além disso, esse novo paradigma educacional se enquadra nas
necessidades e vontades da nova geração, motivando e estimulando a experimentação e
contribuindo para alunos mais autônomos. Permite ainda a interação síncrona e assíncrona até
quando não está on-line e aumenta o contato do professor com os alunos e dos alunos entre si. Os telefones celulares estão cada vez mais acessíveis, modernos e completos. É
necessário destacar a sua portabilidade, interatividade, conectividade e individualidade, além dos
sensores ao contexto, como GPS, que agora são embutidos nos aparelhos celulares. Essa ferramenta
também apresenta diversas funcionalidades, como câmeras fotográficas e de vídeo, gravadores e
reprodutores de som, editores de texto – possibilitando a criação e edição de textos - e inúmeras
outras funções que corroboram a potencialidade da sua aplicação no ensino. Porém, para aplicar um novo possível modelo educacional, devemos estar conscientes 104 http://periodicos.letras.ufmg.br/index.php/textolivre
Ano: 2014 – Volume: 7 – Número: 1 também de certas fraquezas e desvantagens. No caso da aprendizagem móvel, a grande variedade de
plataformas, dispositivos e tecnologias dificulta a unificação e a padronização do processo. É
necessário muito tempo de pesquisa, pois a atualização tecnológica é muito rápida e de complicado
acompanhamento. Além disso, existe a acomodação por parte de alguns professores, provocando a
resistência a essas novas tecnologias. De todas as formas, devemos refletir sobre caminhos que proporcionem melhoras no
ensino e na maneira como os alunos aprendem. CONSIDERAÇÕES FINAIS É necessário levantar questionamentos e discutir
com os educadores métodos adequados e maneiras de realizar formação contínua. Se a tendência
mundial é sempre se desenvolver e evoluir, e percebemos que as novas gerações e as novas
tecnologias estão em constante acompanhamento dessa tendência, devemos também nos
desenvolver e evoluir. O telefone celular surge como uma possibilidade, entre tantas, de tornar o
ensino mais interessante e eficiente. Cabe, então, analisar essa possibilidade e decidir adotá-la – ou
não. REFERÊNCIAS ATTEWELL, J. From research and development to mobile learning: tools for education and
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Ano: 2014 – Volume: 7 – Número: 1 5, out. 2001. Disponível em <http://www.marcprensky.com/writing/Prensky%20-%20Digital
%20Natives,%20Digital%20Immigrants %20-%20Part1.pdf>. Acesso em: 10 jul. 2011. PRENSKY, M. What can you learn from a cell phone? Almost anything! Innovate, v. 1, n. 5,
Florida, 2005. Disponível em: <http://www.innovateonline.info/index.php?view=article&id=83>. Acesso em: 10 jul. 2011. QUINN, C. Mobile, Wireless, In-Your-Pocket Learning. (2000). Disponível em:
<http://learning.ericsson.net/mlearning2/project_one/resources/articles.html>. Acesso em: 24 ago. 2011. RIBEIRO, A.; FERREIRA, E. & CARNEIRO, N. Mobile Learning - As tecnologias telemáticas e a
aprendizagem. Nov. 2011. Disponível em: <http://www.slideshare.net/ElisabeteFerreira/mobile-
learning-mestradomultimedia/>. Acesso em: 03 jan. 2012. SACCOL, A.; SCHLEMMER, E. & BARBOSA, J. M-learning e u-learning: novas perspectivas da
aprendizagem móvel e ubíqua. São Paulo: Pearson Prentice Hall, 2011. SCHLEMMER, E.; SACCOL, A.; BARBOSA, J. & REINHARD, N. M-learning ou aprendizagem
com mobilidade:
casos no contexto brasileiro. (2007). Disponível em:
<http://www.abed.org.br/congresso2007/tc/552007112411PM.pdf>. Acesso em: 26 ago. 2011. SHARPLES, M.; TAYLOR, J. & VAVOULA, G. Theory of learning for the mobile age. ANDREWS, R. & HAYTHORNTHWAITE, C. (Eds.) The SAGE Handbook of E-learning
Research. Londres: SAGE, 2007, pp. 221-247. TELECO. Estatísticas de celulares no Brasil. (2014). Disponível em:
<http://www.teleco.com.br/ncel.asp>. Acesso em: 20 maio 2014. TRAXLER, J. Learning in a mobile age. i
É a qualidade de um objeto, ou de um ambiente, que permite que um indivíduo realize uma ação. ii
Termo utilizado por Prensky (2001) para designar os alunos de hoje, que são todos “falantes nat
digital dos computadores, videogames e internet. ado por Prensky (2001) para designar os alunos de hoje, que são todos “falantes nativos” da língua
omputadores, videogames e internet. i
É a qualidade de um objeto, ou de um ambiente, que permite que um indivíduo realize uma ação.
ii
Termo utilizado por Prensky (2001) para designar os alunos de hoje, que são todos “falantes nativos” da língua
digital dos computadores, videogames e internet. REFERÊNCIAS International Journal of Mobile and Blended Learning, v. 1, n. 1, jan./mar. 2009, pp. 1-12. Disponível em: <http://wlv.academia.edu/JohnTraxler/
Papers/83099/Learning_in_a_Mobile_Age>. Acesso em: 04 jan. 2012. WEISER, M. The computer for the 21st century. Scientific American Special Issue on
Communications, Computers, and Networks, set. 1991. Disponível em: <http://www.ubiq. com/hypertext/weiser/SciAmDraft3.html>. Acesso em: 27 out. 2011. 106
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Genotype by Seeding Rate Interaction in Wheat
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Alexandra Azevedo
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Genotype by Seeding Rate Interaction in Wheat
Genotype by Seeding Rate Interaction in Wheat Follow this and additional works at: https://newprairiepress.org/kaesrr Follow this and additional works at: https://newprairiepress.org/kaesrr
Part of the Agronomy and Crop Sciences Commons Follow this and additional works at: https://newprairiepress.org/kaesrr Part of the Agronomy and Crop Sciences Commons Part of the Agronomy and Crop Sciences Commons ansas Agricultural Experiment Station Research Reports
ansas Agricultural Experiment Station Research Reports ansas Agricultural Experiment Station Research Reports
ansas Agricultural Experiment Station Research Reports Volume 3
Issue 6 Kansas Field Research Article 30 This wheat is available in Kansas Agricultural Experiment Station Research Reports: https://newprairiepress.org/
kaesrr/vol3/iss6/30 This wheat is available in Kansas Agricultural Experiment Station Research Reports: https://newprairiepress.org/
kaesrr/vol3/iss6/30 Recommended Citation
Recommended Citation Azevedo, A. J.; Varela, S.; Lollato, R.; and Ciampitti, I. A. (2017) "Genotype by Seeding Rate Interaction in
Wheat," Kansas Agricultural Experiment Station Research Reports: Vol. 3: Iss. 6. https://doi.org/10.4148/
2378-5977.7447 This report is brought to you for free and open access by New
Prairie Press. It has been accepted for inclusion in Kansas
Agricultural Experiment Station Research Reports by an
authorized administrator of New Prairie Press. Copyright 2017
the Author(s). Contents of this publication may be freely
reproduced for educational purposes. All other rights reserved. Brand names appearing in this publication are for product
identification purposes only. No endorsement is intended, nor
is criticism implied of similar products not mentioned. K-State
Research and Extension is an equal opportunity provider and
employer. This wheat is available in Kansas Agricultural Experiment Station Research Reports: https://newpra
k Summary y
Genotype by seeding rate interaction can play a critical role in understanding wheat
(Triticum aestivum L.) yield potential. The objective of this study was to quantify wheat
yield response to seeding rates by contrasting genotypes (high- vs. low-tillering). One
study was planted at two locations: Ashland Bottoms (dryland and conventional till
age) and at Topeka (irrigated and no-tillage) field research stations (Kansas). The two
winter wheat varieties were sown at four different seeding rates (40, 80, 120, and 160
lb/a). Measurements consisted of stand counts, canopy coverage (estimated via imagery
collection), determination of early-season gaps in the final stand (missing plants), spac
ing between plants, and imagery collection via small-unmanned aerial vehicle systems
(sUAVS). For the first year, average yield was greater at Ashland Bottoms (79.8 bush
els per acre) than at Topeka (50.4 bushels per acre). Statistically, neither seeding rate,
variety, nor their interaction resulted in significant differences at Topeka. Seeding rate
significantly affected yields at Ashland Bottoms, with positive yield response as seeding
rate increased but plateauing at 120 lb/a. Further research is in place to test the geno
type and seeding rate interactions for providing a better understanding of the complex
ity behind the influence of these factors on wheat yields. Genotype by Seeding Rate Interaction
in Wheat A.J. Azevedo, S. Varela, R.P. Lollato, and I.A. Ciampitti Introductionh The interaction between genotype and seeding rate can play a critical role in under
standing wheat yield potential. Genotype differential ability in tillering can affect the
individual plant response to the use of above- and below-ground resources. Addition
ally, the interaction of genotype with seeding rate can affect the tillering response and
final yield considerably. The main goal of the study was to evaluate early-season plant
uniformity and quantify wheat yield response to the interaction of seeding rates by
genotypes with contrasting tillering abilities. Kansas State University Agricultural Experiment Station and Cooperative Extension Service Emergence g
Emergence was summarized at both sites and only for the four seeding rates evaluated in
this study (Figure 3), presenting an expected progression, without portraying significant
differences between treatments. Soil Test Soil test prior to planting at both sites consisted of fifteen samples with ten cores each
(0-6 inches soil depth). At Ashland, soil pH averaged 6.3, organic matter (OM) 2.3%,
Mehlich-3 soil phosphorus (P) 11.2 ppm, and potassium (K) level of 250 ppm. At To
peka, initial soil pH was 7.0, OM 2.8%, Mehlich-3 P 40 ppm, and K 155 ppm. Procedures
T The study was conducted at two sites, 1) Ashland Bottoms (39° 07’ 34” N, 96° 38’ 08”
W), Figure 1 (A), in a Wymore silty clay soil loam (fine, smectitic, mesic Aquertic Argi
udolls), in conventional tillage and preceded by soybeans (Glycine max L.); and 2) To
peka (39° 04’ 35” N, 95° 46’ 04” W), Figure 1 (B), in a Eudora silt loam and sandy loam
(coarse-silty, mixed, superactive, mesic Fluventic Hapludolls), in a no-tillage situation,
fully irrigated and also preceded by soybeans as a previous crop in the rotation scheme. Following common agronomic practices for the region, planting date was October 5
for Ashland Bottoms and October 15 for Topeka. The same procedure was followed to Kansas State University Agricultural Experiment Station and Cooperative Extension Service 1 Kansas Field Research 2017 Kansas State University Agricultural Experiment Station and Cooperative Extension Service Kansas Field Research 2017 control weeds and pests during the growing season. In both sites, nitrogen (N) fertiliza
tion was performed using urea and ammonium nitrate (UAN) (28-0-0) to reach a final
N rate of 50 lb N/a during the completion of tillering (Feekes 3 stage). Treatments
consisted of two different genotypes, WB-Cedar (high tillering) and WB4458 (low
tillering), and four seeding rates (40, 80, 120, and 160 lb/a) with a total of eight treat
ments, as shown in Table 1, and five repetitions. Plots in each location were 80 feet long
by 15 feet wide. Measurements consisted of stand count, percent green canopy coverage estimated via
digital imagery, within-row gap length (missing plants in the stand), plant biomass, and
imagery collected via small unmanned aerial vehicle systems (sUAVS). Speed and uni
formity of plant emergence were measured in three replications in each location using
a new methodology developed for this study. This methodology consisted of selecting
two middle rows and establishing eight linear feet in each plot (Figure 2), from which
all emergence measurements were collected at two-day intervals within the established
areas. Gap measurements were collected at Feekes GS 2, around four to five weeks after plant
ing in order to evaluate the occurrence and pattern of gaps in the final stand. For this
specific measurement, thirty linear feet were established in the middle of each plot. Gap
evaluations were collected from two out of all 8 treatment combinations (80 lb/a WB-
Cedar; 80 lb/a WB4458) and six replications. The gap analysis methodology consisted
of extending a measuring tape in the ground beside each individual row, counting final
stand, and determining the exact geo-position of each gap relative to the beginning of
the plot. A gap was considered and measured when the space between plants was larger
than 4 inches. Biomass Aboveground biomass was collected four times after winter dormancy in the selected
subplots in which emergence and gap analysis were performed. Biomass samples were
obtained from 8 linear feet and then the samples were oven-dried at 60°C until con
stant weight was attained. Plant dry biomass did not show critical differences between
evaluated treatments (Figure 5), but as opposed to the emergence process, larger bio
mass was accumulated at Ashland Bottoms compared to the site at Topeka. Kansas Field Research 2017 Kansas Field Research 2017 more gaps (Figures 4A and B) than the ground-truthing characterization; therefore, the
imagery method needs to be yet perfected in order to classify small plants. more gaps (Figures 4A and B) than the ground-truthing characterization; therefore, the
imagery method needs to be yet perfected in order to classify small plants. Gap Analysish p
y
The gap analysis in the wheat stand provides a better understanding of plant uniformity. This ground-truthing characterization of spacing and uniformity was later implemented
to correlate with the imagery collection data from the sUAVS. Multi-spectral imagery
data, with infrared (IR), near infrared (NIR), red, blue, and green bands, were collected,
and a spectral-band characterization was implemented to differentiate plants from soil
and previous crop residue. Gap analysis helped to relate the pattern of gaps in imagery
with the use of machinery and environmental issues. The imagery method showed many Kansas State University Agricultural Experiment Station and Cooperative Extension Service 2 2 Grain Yields Final yield for both locations did not present a statistical difference between treat
ments. Overall yield for Ashland Bottoms (79.8 bu/a) was greater than at Topeka (50.4
bu/a). Better yields at Ashland Bottoms are the reflection of better reproductive grow
ing conditions. The difference in environments might be explained, in part, due to low
temperature damage that occurred at Topeka during the late winter period. At the Ashland Bottoms site, smaller yield differences were observed for the seeding
rate factor. Notwithstanding, the Ashland Bottoms site presented superior yields and
a clear trend between treatments, seeding rate factor increased, large yield variability
was also recorded in this location. This can be graphically observed through the range
of values for the different treatments in each location (Figure 6). Significant difference
was reported neither for genotype by seeding rate interaction nor for single effect of the
factors evaluated at the Topeka. At Topeka, not only was the overall yield lower but also less yield variability was depict
ed when compared to Ashland Bottoms. Table 1. Experimental factors (genotype and seeding rate combinations) evaluated
Treatments
Factors
1
2
3
4
5
6
7
8
Genotype
WB-Cedar
WB-Cedar
WB-Cedar
WB-Cedar
WB4458
WB4458
WB4458
WB4458
Seeding rate lb/a
40
80
120
160
40
80
120
160
Planting time was during the first two weeks of October in both sites (Ashland Bottoms and Topeka, KS). Table 1. Experimental factors (genotype and seeding rate combinations) evaluated
Treatments
Factors
1
2
3
4
5
6
7
8
Genotype
WB-Cedar
WB-Cedar
WB-Cedar
WB-Cedar
WB4458
WB4458
WB4458
WB4458
Seeding rate lb/a
40
80
120
160
40
80
120
160
Planting time was during the first two weeks of October in both sites (Ashland Bottoms and Topeka, KS). Table 1. Experimental factors (genotype and seeding rate combinations) evaluated Kansas State University Agricultural Experiment Station and Cooperative Extension Service 3 Kansas Field Research 2017 Riley County, KS
Jeferson County, KS
a
b
Figure 1. Winter wheat study located at (a) Ashland Bottoms and (b) Topeka, KS. Riley County, KS
a Figure 1. Winter wheat study located at (a) Ashland Bottoms and (b) Topeka, KS. 2 rows, 8 linear ft
Figure 2. Stand count measurements collected in the two middle rows in eight linear feet
in each plot. Figure 2. Stand count measurements collected in the two middle rows in eight linear feet
in each plot. ansas State University Agricultural Experiment Station and Cooperative Extension Service Kansas State University Agricultural Experiment Station and Cooperative Extension Service Grain Yields 300
200
100
0
Plants emerged
80
60
0
40
20
40 lb/a
20
0
60
40
Days after planting
200
100
0
80 lb/a
120 lb/a
160 lb/a
Ashland Bottoms
Topeka 300
200
100
0
Plants emerged
80
60
0
40
20
40 lb/a
20
0
60
40
Days after planting
200
100
0
80 lb/a
120 lb/a
160 lb/a
Ashland Bottoms
Topeka
Figure 3. Plant emergence dynamics. Graph showing progression of emergence in an eight
square foot area. Ashland Bottoms
Topeka Figure 3. Plant emergence dynamics. Graph showing progression of emergence in an eight
square foot area. Kansas State University Agricultural Experiment Station and Cooperative Extension Service Kansas State University Agricultural Experiment Station and Cooperative Extension Service 4 Kansas Field Research 2017 A
B
Figure 4. Gap analysis, (a) comparison ground truth, (b) imagery, and (c) calibration of
imagery. Figure 4. Gap analysis, (a) comparison ground truth, (b) imagery, and (c) calibration of
imagery. Kansas State University Agricultural Experiment Station and Cooperative Extension Service Kansas State University Agricultural Experiment Station and Cooperative Extension Service 5 5 Kansas Field Research 2017
1,500
1,000
500
0
Plant biomass, lb/a
280
260
240
220
200
180
a
1,500
1,000
500
0
Plant biomass, lb/a
Days after planting
300
250
200
150
b
Figure 5. Plant biomass in lb/a at varying stages, in (a) Ashland Bottoms and (b) Topeka. Kansas Field Research 2017 1,500
1,000
500
0
Plant biomass, lb/a
280
260
240
220
200
180
a Plant biomass, lb/a 280 1,500
1,000
500
0
Plant biomass, lb/a
Days after planting
300
250
200
150
b
Figure 5. Plant biomass in lb/a at varying stages, in (a) Ashland Bottoms and (b) Topeka. Plant biomass, lb/a Days after planting Figure 5. Plant biomass in lb/a at varying stages, in (a) Ashland Bottoms and (b) Topeka. Kansas State University Agricultural Experiment Station and Cooperative Extension Service 6 6 Kansas Field Research 2017
1,500
1,000
500
0
Grain yield, bu/a
8
7
6
3
2
1
a
1,500
1,000
500
0
Grain yield, bu/a
Treatments
b
5
4
8
7
6
3
2
1
5
4
Figure 6. Grain yield, 13.5% moisture, for all systems evaluated at harvest time at (a) Ash
land Bottoms and (b) Topeka sites, KS (2015-16 growing season). Kansas Field Research 2017 1,500
1,000
500
0
Grain yield, bu/a
8
7
6
3
2
1
a
5
4 Figure 6. Grain Yields Grain yield, 13.5% moisture, for all systems evaluated at harvest time at (a) Ash
land Bottoms and (b) Topeka sites, KS (2015-16 growing season). Kansas State University Agricultural Experiment Station and Cooperative Extension Service
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Various open science
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Queratitis herpética atípica multifocal secundaria a corticoides tópicos
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Spanish
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Virgilio Galvis, Alejandro Tello, Lisi Rodríguez, Lizeth
Ardila, Angélica Pedraza-Concha, Camilo Niño
Revista de la Facultad de Ciencias de la Salud, Universidad Autónoma de Bucaramanga (Colombia)
Vol. 22. Número 1 – abril – julio 2019
revista de la facultad
de ciencias de la salud
i-ISSN 0123-7047 e-ISSN 2382-4603 https://doi.org/10.29375/issn.0123-7047
Sensation Seeking
Anticoagulantes
Obra titulada “Saline Infusion: An incident in the British Red Cross Hospital, Arc-en-Barrois, 1915”, 1915 por: Henry Tonks (1862-1937)
Cáncer Colorrectal
Imágenes de medicina clínica
Vol. 22(1):12-15, abril – julio 2019
Queratitis herpética atípica multifocal secundaria
a corticoides tópicos
Multifocal atypical herpetic keratitis secondary to topical corticosteroids
Ceratite herpética multifocal atípica secundária a corticoides tópicos
Virgilio Galvis, MD., Esp., PhD.1, Alejandro Tello, MD., Esp., PhD.1 , Lisi
Rodríguez, MD.2, Lizeth Ardila, MD.3, Angelica Pedraza-Concha, MD.4 ,
Camilo Niño, MD., Esp.5
1. Médico, Especialista en Oftalmología, Subespecialista en Facoemulsificación y Segmento Anterior,
Subespecialista en Cirugía refractiva, Doctorado en Investigación, Cirugía y especialidades Médicoquirúrgicas, Doctorado en Ciencias de la Visión, Centro Oftalmológico Virgilio Galvis, Universidad
Autónoma de Bucaramanga, Fundación Oftalmológica de Santander, Floridablanca, Santander, Colombia.
2. Médico, Universidad Autónoma de Bucaramanga, Floridablanca, Santander, Colombia
3. Médico, Médico de Bienestar Universitario, Universidad Autónoma de Bucaramanga, Bucaramanga,
Santander, Colombia
4. Médico, en Servicio Social Obligatorio, Asistente Editorial, Universidad Autónoma de Bucaramanga,
Floridablanca, Santander, Colombia
5. Médico, Especialista en Oftalmología, Subespecialista en Córnea y Segmento Anterior. Centro
oftalmológico Virgilio Galvis, Fundación Oftalmológica de Santander, Universidad Autónoma de
Bucaramanga, Floridablanca, Santander, Colombia.
Correspondencia: Angélica Pedraza Concha, Calle 157 No. 14 - 55, Floridablanca, Santander, Colombia.
E-mail: apedraza72@unab.edu.co
Cómo citar: Galvis V, Tello A, Rodríguez L, Ardila L, Pedraza-Concha A, Niño C. Queratitis herpética atípica
multifocal secundaria a corticoides tópicos. MedUNAB. 2019;22(1):12-15. doi: 10.29375/01237047.3637
INFORMACIÓN ARTÍCULO
Artículo recibido: 25 de mayo de 2019
Artículo aceptado: 10 de julio de 2019
DOI: https://doi.org/10.29375/01237047.3637
12
Virgilio Galvis, Alejandro Tello, Lisi Rodríguez, Lizeth Ardila, Angélica Pedraza-Concha, Camilo Niño
DOI: https://doi.org/10.29375/01237047.3637
Vol. 22(1):12-15, abril – julio 2019
Figura A. OI: fotografía en lámpara de hendidura con filtro de azul de cobalto. La tinción con fluoresceína permite
apreciar dos úlceras separadas de aspecto dendrítico (flechas blancas) que comprometen la media periferia, tanto
superonasal (meridiano de las 9 y 10:30 horas), como superior (meridiano de las 12 y las 2 horas). Adicionalmente,
lesión ulcerativa con compromiso estromal de aproximadamente de 3x2 mm con adelgazamiento de más del 50 %
del espesor corneal (óvalo blanco). Figura B. OI: tomografía de coherencia óptica de la córnea. Se observa área de
adelgazamiento (remanente estromal respetado de 171 μm) con quiste epitelial sobre el área de ulceración estromal.
Figura C. OI: fotografía en lámpara de hendidura con filtro de azul de cobalto. La tinción de fluoresceína permite
apreciar solo mínima captación del colorante sobre las áreas de las úlceras dendríticas epiteliales, así como cicatrización
casi completa de la ulceración estromal. Figura D. OI: tomografía de coherencia óptica de la córnea. Epitelización
completa del área adelgazada (que alcanza ahora 276 μm de espesor) con desaparición casi total del quiste epitelial.
Figure A. OS: Slit-lamp biomicroscopy photograph with Cobalt Blue Filter. Fluorescein staining reveals two separate
dendritic ulcers (white arrows) compromising the mid-periphery, both superonasal (at 9 and 10:30 o’clock), and
superior (at 12 and 2 o’clock). Additionally, an ulcerative lesion with stromal compromise of about 3x2 mm with
thinning of more than 50% of the total corneal thickness (white oval). Figure B. OS: optical coherence tomography
of the cornea. An area of thinning (respected stromal remnant of 171 μm) with epithelial cyst on the stromal ulceration
area. Figure C. OS: Slit-lamp biomicroscopy photograph with Cobalt Blue Filter. Fluorescein stain shows minimal
staining on areas of epithelial dendritic ulcers, as well as almost complete healing of the stromal ulceration. Figure
D. OS: optical coherence tomography of the cornea. Complete epithelialization of the thinned area (now reached
thickness of 276 μm) and almost complete resolution of the epithelial cyst.
Figura A. OE: fotografia com lâmpada de fenda com filtro azul cobalto. A coloração com fluoresceína revela duas
úlceras dendríticas (setas brancas) que comprometem a periferia média, tanto superonasal (meridiano das 9 e 10:30
horas), quanto superior (meridiano das 12 e das 14 horas). Além disso, lesão ulcerativa com comprometimento do
estroma de aproximadamente de 3x2 mm com afilamento de mais de 50% da espessura corneal (oval branco). Figura
B. OE: tomografia de coerência óptica (OCT) de córnea. Observa-se uma área afinada (restos de estroma respeitados
de 171 μm) com cisto epitelial sobre a área de ulceração do estroma. Figura C. OE: fotografía em lâmpada de fenda
com filtro azul cobalto. A coloração com fluoresceína permite avaliar apenas a captação mínima do corante nas
áreas das úlceras dendríticas epiteliais, bem como cicatrização quase completa da ulceração estromal. Figura D.
OE: tomografía de coerência óptica (OCT) de córnea. Epitelização completa da área afinada (que agora atinge 276
μm de espessura) com quase total desaparecimento do cisto epitelial.
13
Queratitis herpética atípica multifocal secundaria a corticoides tópicos
Hombre de 53 años, sin antecedentes de importancia,
con cuadro clínico de 30 días de evolución caracterizado
por dolor tipo punzada y sensación de cuerpo extraño en
ojo izquierdo (OI). Por indicación de un farmacéutico,
usó colirio de dexametasona, neomicina, polimixina B
(Wassertrol®, Wassser) 3 veces al día durante 15 días,
con empeoramiento de los síntomas, motivo por el cual
consultó al Centro Oftalmológico. Al examen físico, se
encontraron úlceras dendríticas epiteliales en periferia
superior y superonasal; además, otra úlcera de 2.5 mm de
diámetro con compromiso estromal, con adelgazamiento
en el área de la lesión y quistes epiteliales. El cuadro se
consideró como afectación ocular por el virus del herpes
expresada en una forma clínica de queratitis epitelial
infecciosa (morfología dentrítica múltiple) y, además,
queratitis estromal. Esta evolución atípica de la queratitis
herpética (Figuras A y B) se relaciona, posiblemente,
con el previo uso de esteroides tópicos. Se suspendió el
colirio triconjugado y se inició manejo con ganciclovir
gel oftálmico tópico 5 veces al día y aciclovir 2 g diarios
vía oral, con buena respuesta evidenciada en el control al
séptimo día (Figuras C y D).
tratamiento tópico con esteroides. Cuando se sospeche
infección por VHS, los esteroides tópicos mal indicados
pueden generar empeoramiento del cuadro clínico (1),
como en el caso expuesto.
Para el examen del detalle de las lesiones corneales,
se requieren una lámpara de hendidura y la tinción
con fluoresceína, siendo improbable que un médico no
oftalmólogo las pueda observar. Por ello, se debe basar
en la evolución clínica y en los hallazgos macroscópicos
para enfocar el diagnóstico y manejo. Entre las claves
para diferenciar la conjuntivitis de la queratitis epitelial
herpética están la presencia de secreción mucopurulenta
en la conjuntivitis bacteriana, ausencia de fotofobia, y
usualmente bilateral (ya sea bacteriana o adenoviral). En
la queratitis epitelial herpética por su parte, se presenta
secreción acuosa, hay fotofobia, usualmente unilateral
y hay disminución de la sensibilidad corneal (1).
Conflictos de interés
Los autores declaran no tener ningún conflicto de
interés.
El virus del herpes simplex (VHS) tipo I es una frecuente
causa de queratitis (1), que puede comprometer cualquier
capa de la córnea (epitelio, estroma o endotelio), aunque
su presentación epitelial es la más común (50 % - 80 %
de los casos de herpes ocular) (2).
Referencias
1. Seitz B, Heiligenhaus A. “Herpeskeratitis” Unterschiedliche Ausprägungsformen erfordern unterschiedliche Therapieansätze. Ophthalmologe.
2011;108(4): 385-97. doi: 10.1007/s00347-011-2346-5
2. Wilhelmus KR. Antiviral treatment and other
therapeutic interventions for herpes simplex virus epithelial keratitis. Cochrane Database Syst
Rev. 2015;1:CD002898. doi: 10.1002/14651858.
CD002898.pub5.
3. Thygeson P, Hogan MJ, Kimura SJ. The unfavorable
effect of topical steroid therapy on herpetic keratitis.
Trans Am Ophthalmol Soc [Internet]. 1960 [citado
20 de mayo de 2019];58:245-62. Recuperado a partir de: https://www.ncbi.nlm.nih.gov/pmc/articles/
PMC1316376/
4. Patterson A, Jones BR. The management of ocular herpes. Trans Ophthalmol Soc UK [Internet]. 1967 [citado 20 de mayo de 2019];87:59-84. Recuperado a partir
de: https://www.ncbi.nlm.nih.gov/pubmed/4892489
5. Patterson A. Management of ocular herpes simplex.
Br J Ophthalmol. 1967;51(7):494-5. doi: 10.1136/
bjo.51.7.494
6. Roozbahani M, Hammersmith KM. Management
of herpes simplex virus epithelial keratitis. Curr
Opin Ophthalmol. 2018;29(4):360-4. doi: 10.1097/
ICU.0000000000000483.
Desde hace más de 50 años se conoce la contraindicación
de los corticosteroides si existe compromiso epitelial
corneal por VHS, pues aumentan el riesgo de que la
úlcera progrese de su forma dendrítica a una geográfica,
así como la posibilidad de multifocalidad de las lesiones
(1,3-5) e, incluso, se incrementa la eventualidad de
compromiso estromal, como en el presente caso (3-7).
Ante la sospecha de conjuntivitis infecciosa, el médico
no oftalmólogo podría abordar el caso con colirios
de antibióticos tópicos. Si en 48 horas no mejoran
los síntomas o aparecen nuevos síntomas de alarma
(fotofobia, disminución de la visión o dolor) se debe
remitir al oftalmólogo. Por otra parte, el uso de esteroides
tópicos debe estar restringido, ya que puede agravar
la afectación ocular por el virus del herpes, como en
este caso. A pesar de que los esteroides tópicos pueden
ser usados para el tratamiento de diversas condiciones
que se manifiestan con ojo rojo, sus indicaciones son
muy precisas (8). Desafortunadamente, en nuestro
medio son usados inapropiadamente tanto por médicos
generales, como por los mismos pacientes siguiendo
indicaciones de personal no médico (farmacéuticos),
como en el presente caso. La recomendación es evaluar
cuidadosamente al paciente antes de comenzar un
14
Virgilio Galvis, Alejandro Tello, Lisi Rodríguez, Lizeth Ardila, Angélica Pedraza-Concha, Camilo Niño
DOI: https://doi.org/10.29375/01237047.3637
7. Prakash G, Avadhani K, Srivastava D. The three faces
of herpes simplex epithelial keratitis: a steroid-induced
situation. BMJ Case Rep. 2015;2015:bcr2014209197.
doi: 10.1136/bcr-2014-209197
8. Galvis V, Tello A, Diaz CA. Diagnóstico del ojo rojo
para el médico de atención primaria. MedUNAB [Internet]. 2008 [citado 20 de mayo de 2019];11:231-8.
Recuperado a partir de https://revistas.unab.edu.co/
index.php/medunab/article/view/66.
15
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Tumor Suppressor 4.1N/EPB41L1 is Epigenetic Silenced by Promoter Methylation and MiR-454-3p in NSCLC
|
Qin Yang
|
English
|
Spoken
| 7,988
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ORIGINAL RESEARCH
published: 20 June 2022
doi: 10.3389/fgene.2022.805960 ORIGINAL RESEARCH
published: 20 June 2022
doi: 10.3389/fgene.2022.805960 Edited by:
Biaoru Li, Edited by:
Biaoru Li,
Augusta University, United States Edited by:
Biaoru Li,
Augusta University, United States
Reviewed by:
Fei Han,
Army Medical University, China
Hongde Liu,
Southeast University, China
*Correspondence:
Jing Liu
jingliucsu@hotmail.com
Wen Zou
zouwen29w@csu.edu.cn
Hui Li
lihuiscience@163.com
†These authors contributed equally to
this work Non–small-cell lung cancer (NSCLC) is divided into three major histological types, namely, lung
adenocarcinoma (LUAD), lung squamous cell carcinoma (LUSC), and large-cell lung carcinoma
(LCLC). We previously identified that 4.1N/EPB41L1 acts as a tumor suppressor and is
reduced in NSCLC patients. In the current study, we explored the underlying epigenetic
mechanisms of 4.1N/EPB41L1 reduction in NSCLC. The 4.1N/EPB41L1 gene promoter
region was highly methylated in LUAD and LUSC patients. LUAD patients with higher
methylation level in the 4.1N/EPB41L1 gene promoter (TSS1500, cg13399773 or TSS200,
cg20993403) had a shorter overall survival time (Log-rank p = 0.02 HR = 1.509 or Log-rank p =
0.016 HR = 1.509), whereas LUSC patients with higher methylation level in the 4.1N/EPB41L1
gene promoter (TSS1500 cg13399773, TSS1500 cg07030373 or TSS200 cg20993403) had
a longer overall survival time (Log-rank p = 0.045 HR = 0.5709, Log-rank p = 0.018 HR = 0.68
or Log-rank p = 0.014 HR = 0.639, respectively). High methylation of the 4.1N/EPB41L1 gene
promoter appeared to be a relatively early event in LUAD and LUSC. DNA methyltransferase
inhibitor 5-Aza-2′-deoxycytidine restored the 4.1N/EPB41L1 expression at both the mRNA
and protein levels. MiR-454-3p was abnormally highly expressed in NSCLC and directly
targeted 4.1N/EPB41L1 mRNA. MiR-454-3p expression was significantly correlated with
4.1N/EPB41L1 expression in NSCLC patients (r = −0.63, p < 0.0001). Therefore, we
concluded that promoter hypermethylation of the 4.1N/EPB41L1 gene and abnormally high
expressed miR-454-3p work at different regulation levels but in concert to restrict 4.1N/
EPB41L1 expression in NSCLC. Taken together, this work contributes to elucidate the
underlying epigenetic disruptions of 4.1N/EPB41L1 deficiency in NSCLC. Abbreviations: 3′UTRs, 3′-untranslated regions; 5-Aza-CdR, 5-Aza-2′-deoxycytidine; HR, hazard ratio; LCLC, large-cell lung
carcinoma; LR, log-likelihood ratio; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; NSCLC, non–
small-cell lung cancer; SD, standard deviation; TBS-seq, targeted bisulfite sequencing; TSS, transcription start site. Tumor Suppressor 4.1N/EPB41L1 is
Epigenetic Silenced by Promoter
Methylation and MiR-454-3p in NSCLC
Qin Yang 1,2†, Lin Zhu 1†, Mao Ye 3,4, Bin Zhang 5, Peihe Zhan 5, Hui Li 1,3,4*, Wen Zou 6* and
Jing Liu 1* Qin Yang 1,2†, Lin Zhu 1†, Mao Ye 3,4, Bin Zhang 5, Peihe Zhan 5, Hui Li 1,3,4*, Wen Zou 6* and
Jing Liu 1* 1Molecular Biology Research Center and Center for Medical Genetics, School of Life Sciences, Central South University,
Changsha, China, 2School of Medical Laboratory, Shao Yang University, Shaoyang, China, 3Molecular Science and Biomedicine
Laboratory, College of Biology, College of Chemistry and Chemical Engineering, Collaborative Innovation Center for Chemistry
and Molecular Medicine, Hunan University, Changsha, China, 4State Key Laboratory for Chemo/Biosensing and Chemometrics,
College of Biology, College of Chemistry and Chemical Engineering, Collaborative Innovation Center for Chemistry and Molecular
Medicine, Hunan Univers ity, Changsha, China, 5Department of Histology and Embryology, Xiangya School of Medicine, Central
South University, Changsha, China, 6Department of Oncology, The Second Xiangya Hospital of Central South University, Central
South University, Changsha, China Keywords: 4.1N/EPB41L1, lung adenocarcinoma (LUAD), methylation, miR-454-3P, non–small-cell lung cancer
(NSCLC), lung squamous cell carcinoma (LUSC) Edited by:
Biaoru Li, Reviewed by:
Fei Han,
Army Medical University, China
Hongde Liu,
Southeast University, China Reviewed by:
Fei Han,
Army Medical University, China
Hongde Liu,
Southeast University, China *Correspondence:
Jing Liu
jingliucsu@hotmail.com
Wen Zou
zouwen29w@csu.edu.cn
Hui Li
lihuiscience@163.com
†These authors contributed equally to
this work †These authors contributed equally to
this work Specialty section:
This article was submitted to
Epigenomics and Epigenetics,
a section of the journal
Frontiers in Genetics Specialty section:
This article was submitted to
Epigenomics and Epigenetics,
a section of the journal
Frontiers in Genetics Received: 31 October 2021
Accepted: 08 April 2022
Published: 20 June 2022 INTRODUCTION Cell Culture
MRC5, 95C, and 95D cells were grown in DMEM medium
(Gibco, United States) and supplemented with 10% fetal
bovine serum (Gibco, United States). H460 and A549 cells
were grown in RPMI 1640 medium (Gibco, United States) and
supplemented with 10% fetal bovine serum. All the cells were
grown at 37°C in a humidified atmosphere containing 5% CO2. Lung cancer is the most commonly diagnosed cancer and the
most lethal cause of cancer mortality worldwide (Bray et al.,
2018). Non–small-cell lung cancer (NSCLC), consisting of lung
adenocarcinoma
(LUAD),
lung
squamous
cell
carcinoma
(LUSC), and large-cell lung carcinoma (LCLC) (Rodriguez-
Canales et al., 2016), represents major types of lung cancers
(80–85%) (D’Addario et al., 2010). Owing to a lack of obvious
early symptoms and early-stage diagnosis, most patients with
NSCLC are diagnosed in the advanced clinical stage—that is—III
or IV (Norouzi and Hardy, 2021). Despite recent advances in
NSCLC treatment, less than 15% of the patients eventually
survived
(Quintanal-Villalonga
and
Molina-Pinelo,
2019;
Norouzi and Hardy, 2021). treatment 5-Aza-CdR (Merck, Germany) was diluted in PBS. The cells were
seeded in a 6-well plate and treated with 0, 1, or 10 μM 5-Aza-
CdR for 48 h. 5-Aza-CdR was replaced every 24 h. Citation: Yang Q, Zhu L, Ye M, Zhang B, Zhan P,
Li H, Zou W and Liu J (2022) Tumor
Suppressor 4.1N/EPB41L1 is
Epigenetic Silenced by Promoter
Methylation and MiR-454-3p
in NSCLC. June 2022 | Volume 13 | Article 805960 Frontiers in Genetics | www.frontiersin.org 1 4.1N/EPB41L1 is Epigenetic Silenced Yang et al. Targeted Bisulfite Sequencing Targeted Bisulfite Sequencing
The cells were sent to Biomarker Acegene Corporation,
Shenzhen, China for targeted bisulfite sequencing (TBS-seq)
analysis. 4.1N/EPB41L1 promoter methylation was assessed
according to the previously published method (Gao et al.,
2014a; Gao et al., 2014b; Gao et al., 2015; Pan et al., 2018). Methylation levels are defined as the fraction of read counts of ‘C’
in the total read counts of both ‘C’ and ‘T’ for each covered C site. On the basis of such read fraction, methylated cytosine was called
using a binomial distribution as in the method described by Lister
et al. (2009), whereby a probability mass function is calculated for
each methylation context (CpG). Two-tailed Fisherʼs exact test
was used to identify cytosines that are differentially methylated
between two samples or groups. Only those CGs covered by at
least 200 reads in at least one sample were considered for testing. Our
previous
studies
suggested
that
4.1N/EPB41L1
is
abnormally low expressed and exerts anticancer effects in
NSCLC (Wang et al., 2016; Yang et al., 2016; Yang et al.,
2021). In the current study, for the first time, we focus on
identifying the underlying epigenetic disruptions of 4.1N/
EPB41L1 deficiency in NSCLC. We report that promoter
hypermethylation
and
aberrant
miR-454-3p
expression
regulate
4.1N/EPB41L1
expression
at
transcriptional
and
posttranscriptional levels, respectively, but work in concert to
restrict its expression in NSCLC. NSCLC Tissue Samples Tumor tissues and tumor-adjacent tissues were obtained from the
Second Xiangya Hospital of Central South University (Changsha,
China). The tissue samples were subjected to qPCR experiments
after approval by the Ethics Committee of the Second Xiangya
Hospital. Informed consent was obtained from all participating
subjects. Norouzi and Hardy, 2021). Gene promoter methylation and miRNA dysregulation are
typical markers of cancer epigenetics (Nebbioso et al., 2018). Gene promoter methylation most commonly occurs at the CpG
islands and regulates the gene expression at the transcriptional
level (Yang et al., 2014; Zhou et al., 2017; Arechederra et al., 2018). 5–10% of CpG islands in the promoter of genes have been
identified as cancer-specifically methylated, which should not
be methylated in normal cells (Heller et al., 2013; Olbromski et al.,
2020). The methylations of certain genes are of clinical relevance
for patients with NSCLC (Heller et al., 2013). MiRNAs are
endogenous small non-coding RNAs, which directly bind to
the 3′-untranslated regions (3′UTRs) of target mRNAs to
regulate the gene expression at the posttranscriptional level. NSCLC patients have widespread dysregulation of miRNA
expression (Du et al., 2018; Uddin and Chakraborty, 2018). It
has been well-documented that 4.1 family members 4.1N/
EPB41L1 and its homologs (4.1B/EPB41L3, 4.1G/EPB41L2, and
4.1R/EPB41) are lost in various cancers (Yang et al., 2021). However, epigenetic silencing of 4.1 family members in
cancers is still largely unknown. Loss of 4.1B/EPB41L3 is the
only case that has been linked to high promoter methylation in
cancers (Kikuchi et al., 2005; Zhang et al., 2012). No miRNAs
have been found to regulate 4.1 family members. Methylation-Based Analysis y
y
The MethSurv tool (https://biit.cs.ut.ee/methsurv/) (Modhukur
et al., 2018) was used to perform the assessment of methylation-
based analysis for the 4.1N/EPB41L1 gene in LUAD and LUSC. The raw data for LUAD and LUSC could be downloaded from the
website (https://biit.cs.ut.ee/methsurv/). Cell Transfection and Western Blot
MiR-454-3p and the control mimics were purchased from
RiboBio (Guangzhou, China) and transfected according to our
previously published protocol (Li et al., 2016). Western blot was
also performed according to our previous protocol (Yang et al.,
2016). Dual-Luciferase Reporter Gene Assay Dual Luciferase Reporter Gene Assay
The 3′UTR target sites of 4.1N/EPB41L1 mRNA were amplified
by PCR with genomic DNA from MRC5 cells. The PCR product
was cloned in the psiCHECK2 vector (Promega, United States) to
construct the wild-type plasmid (psiCHECK2-4.1N-wt). The
corresponding mutant psiCHECK2-4.1N-mut was constructed
by in vitro site-directed mutagenesis (Mut ExpressMultiS Fast
Mutagenesis
Kit,
Vazyme
Biotech,
China). Bidirectional
sequencing was applied to confirm the correct sequence of the
two constructs. For the dual-luciferase reporter gene assay, A549
and H460 cells were cultured in a 24-well plate for 24 h and
transfected with psiCHECK2-4.1N-wt or psiCHECK2-4.1N-mut
plasmids and miR-454-3p mimics or miR-negative-control using
the RiboFECT™CP transfection kit (Ribo Biotechnology, China)
and Lipofectamine 2000 (Invitrogen, United States). 48 hours
after the transfection, the Dual-Luciferase Reporter Assay System
(Promega, Madison, WI, United States) was used to measure the
luciferase activity according to the manufacturer’s protocol. Statistics All the experiments were performed in triplicate, and statistical
analyses were conducted using GraphPad Prism 5.0. The data
were presented as the mean ± standard deviation (SD). Student’s
t-tests were used to calculate the results. A p-value < 0.05 was
considered significant statistically. DNA methylation values were
represented as beta values (range from 0 to 1). Any beta value
equal to or greater than 0.6 was considered fully methylated. Any
beta value equal to or less than 0.2 was considered to be fully
unmethylated. Beta values between 0.2 and 0.6 were considered to
be partially methylated. Differential methylation for individual
CpG loci was assessed by comparing the beta values. The patients
were classified into high-methylation and low-methylation levels
based on maxstat (Modhukur et al., 2018). Cox proportional
hazards models were used to perform the survival analysis based
on methylation levels of the CpG sites. The methylation levels and
overall survival time were used as explanatory variables and
response variables, respectively, to perform overall survival
analysis. Prognostic Relevance of 4.1N/EPB41L1
Hypermethylation in LUAD and LUSC yp
y
In most cases, evaluating DNA methylation signature in the
promoter region is highly desirable and sensitive for cancer
diagnosis and prognosis. The Kaplan–Meier survival curve
showed that the higher methylation levels in the promoter
(TSS1500, cg13399773 or TSS200, cg20993403) of the 4.1N/
EPB41L1 gene were significantly associated with a shorter overall
survival time (Log-rank p = 0.02, HR = 1.509 or Log-rank p = 0.016,
HR = 1.509, respectively) for LUAD patients (Figures 3A,C). Median methylation levels of the two CpG sites (cg13399773 and
cg20993403) were high (beta>0.5) at stage I and did not essentially
change in tumors of more advanced stages (Figures 3B,D). Unlike
the LUAD, the Kaplan–Meier survival curve showed that the higher
methylation levels in the promoter (TSS1500 cg13399773, TSS1500
cg07030373, or TSS200 cg20993403) of the 4.1N/EPB41L1 gene were
significantly associated with a shorter overall survival time (Log-rank
p = 0.045 HR = 0.5709, Log-rank p = 0.018 HR = 0.68 or Log-rank
p = 0.014 HR = 0.639 respectively) for LUSC patients (Figures
4A,C,E). Median methylation levels of the three CpG sites (TSS1500
cg13399773, TSS1500 cg07030373, or TSS200 cg20993403) were
high (beta>0.5) from stage I to IV and tended to decline with tumor
progression (Figures 4B,D,F). We did not further explore the
contrasting
prognostic
relevance
of
4.1N/EPB41L1
hypermethylation
between
LUAD
and
LUSC. However,
methylation of the 4.1N/EPB41L1 gene might be an important
mechanism for tumor formation in LUAD and LUSC because
high 4.1N/EPB41L1 gene methylations were observed at stage I. Because the MethSurv tool (https://biit.cs.ut.ee/methsurv/) lacks
LCLC
data,
the
prognostic
relevance
of
4.1N/EPB41L1
hypermethylation in LCLC was unknown. RNA Extraction and qPCR Rabbit
anti-4.1N
antibody
was
purchased
from
ATLAS
(Bromma,
Sweden). Rabbit
anti-GAPDH
antibody
was
purchased
from
Santa
Cruz
(Santa
Cruz
Biotechnology,
United States). Total RNA was isolated using the RNeasy kit (QIAGEN,
United States). cDNA was synthesized using the RevertAid H
Minus First-Strand cDNA Synthesis Kit (Thermo Scientific, June 2022 | Volume 13 | Article 805960 Frontiers in Genetics | www.frontiersin.org 2 4.1N/EPB41L1 is Epigenetic Silenced Yang et al. CpG islands in the 4.1N/EPB41L1 gene promoter (2,000 bp
upstream to 1,000 bp downstream of the transcription start
site, Figure 1A). NSCLC predominantly encompasses the
LUAD
(40%
prevalence)
and
LUSC
subtypes
(25%
prevalence). The MethSurv tool (https://biit.cs.ut.ee/methsurv/)
(Modhukur et al., 2018) was used to perform the assessment of
methylation-based analysis for the 4.1N/EPB41L1 gene promoter
region (TSS200 and TSS1500) in LUAD and LUSC. The heat map
showed that high methylation of the 4.1N/EPB41L1 gene was
prevalent in both LUAD (Figure 1B) and LUSC (Figure 1C). Because the MethSurv tool (https://biit.cs.ut.ee/methsurv/) lacks
LCLC data, we investigated the methylation of the 4.1N/EPB41L1
gene in LCLC cells (95C, 95D, and H460) and normal lung
fibroblast cells (MRC5). TBS-seq results showed that promoter
methylation of 4.1N/EPB41L1 was significantly higher in LCLC
cells (95C, 95D, and H460) than in normal lung cells (MRC5) (p <
0.001) (Figures 1D,E). To further validate the role of methylation
in 4.1N/EPB41L1 gene repression, we treated the LCLC cells (95C
and H460) and LUAD cells (A549) with DNA methyltransferase
inhibitor 5-Aza-CdR. After demethylation treatment, the 4.1N/
EPB41L1 gene was restored both at mRNA (Figures 2A–C) and
protein levels (Figures 2D–F). Taken together, these results
indicated that 4.1N/EPB41L1 gene methylation is a cause of
decreased 4.1N/EPB41L1 expression in NSCLC patients. United States). Stem-loop RT primers (RiboBio, China) were
used in reverse transcription for miR-454-3p. qPCR was
performed using the One-Step qRT-PCR SYBR® Green Kit
(Vazyme Biotech, China). The sequences of primers targeting
4.1N/EPB41L1 and miR-454-3p were used as described earlier
(Wang et al., 2010) and designed by Vazyme Biotech (Nanjing,
China). U6 small nuclear RNA was used as an internal control for
miR-454-3p analysis. Hypermethylation of the 4.1N/EPB41L1
Gene in NSCLC Aberrant hypermethylations in the promoter region of genes are
considered a major reason for gene silencing in cancer (Lamy
et al., 2001). The CpG island methylation prediction using the
CpGPNP program (http://forensicdna.kr/cpgpnp/) showed four June 2022 | Volume 13 | Article 805960 Frontiers in Genetics | www.frontiersin.org 3 4.1N/EPB41L1 is Epigenetic Silenced Yang et al. Yang et al. FIGURE 1 | Promoter methylation of the 4.1N/EPB41L1 gene in NSCLC. (A) In the upper panel, CpG island prediction of the 4.1N/EPB41L1 gene promoter was
ntegrated using the CpGPNP program. The red line represented GC content, the green line represents CpG O/E value, and the yellow box represented four predicte
ocations of CpG island (range from 82 to 179, 265–361, 2412–2467, and 2542–3002, sequence length >100 bp, GC content >50%, O/E value >0.6). The lower pan
represented the structure of the 4.1N/EPB41L1 gene promoter (range from TSS -2000 to TSS 1000). Coordinate values of abscissas in the lower and upper panels
were correlated. TSS -2000 in the lower panel corresponded to 0 in the upper panel. The TSS site in the lower panel corresponded to 2000 in the upper panel. TSS 100
n the lower panel corresponded to 3000 in the upper panel. (B,C) Heatmap depicting the CpG methylation level of the 4.1N/EPB41L1 gene promoter in LUAD and LUS
patients. Rows and columns represented the CpGs and the patients, respectively. (D) Heatmap depicting the methylation levels of the 4.1N/EPB41L1 gene promoter
normal lung fibroblast cells MRC5 and LCLC cells (95C, 95D, and H460). The row label was the methylation site. The number of row labels corresponded to th
coordinate value of abscissa in the upper panel of Panel 1A. Methylation levels were represented as beta values and shown as a continuous variable from blue to red. A
(Continued FIGURE 1 | Promoter methylation of the 4.1N/EPB41L1 gene in NSCLC. (A) In the upper panel, CpG island prediction of the 4.1N/EPB41L1 gene promoter was
integrated using the CpGPNP program. The red line represented GC content, the green line represents CpG O/E value, and the yellow box represented four predicted
locations of CpG island (range from 82 to 179, 265–361, 2412–2467, and 2542–3002, sequence length >100 bp, GC content >50%, O/E value >0.6). The lower panel
represented the structure of the 4.1N/EPB41L1 gene promoter (range from TSS -2000 to TSS 1000). Coordinate values of abscissas in the lower and upper panels
were correlated. Hypermethylation of the 4.1N/EPB41L1
Gene in NSCLC TSS -2000 in the lower panel corresponded to 0 in the upper panel. The TSS site in the lower panel corresponded to 2000 in the upper panel. TSS 1000
in the lower panel corresponded to 3000 in the upper panel. (B,C) Heatmap depicting the CpG methylation level of the 4.1N/EPB41L1 gene promoter in LUAD and LUSC
patients. Rows and columns represented the CpGs and the patients, respectively. (D) Heatmap depicting the methylation levels of the 4.1N/EPB41L1 gene promoter in
normal lung fibroblast cells MRC5 and LCLC cells (95C, 95D, and H460). The row label was the methylation site. The number of row labels corresponded to the
coordinate value of abscissa in the upper panel of Panel 1A. Methylation levels were represented as beta values and shown as a continuous variable from blue to red. Any
(Continued) Frontiers in Genetics | www.frontiersin.org June 2022 | Volume 13 | Article 805960 4.1N/EPB41L1 is Epigenetic Silenced Yang et al. FIGURE 1 | beta value equal to or greater than 0.6 was considered fully methylated. Any beta value equal to or less than 0.2 was considered to be fully unmethylated. Beta values between 0.2 and 0.6 were considered to be partially methylated. (E) Boxplots indicating the methylation differences between normal lung fibroblast cells
MRC5 and LCLC cells (95C, 95D, and H460). Median methylation levels (show by a thick black line). ***p < 0.001. 4.1N/EPB41L1 mRNA is a Direct Target of
miR-454-3p abolished by the mutation in the miR-454-3p-binding region of
the 4.1N/EPB41L1 mRNA 3′UTR in H460 and A549 cells
(Figures 5G,H). The abovementioned results signified that
abnormally highly expressed miR-454-3p is another epigenetic
cause of 4.1N/EPB41L1 decreasing in NSCLC patients. p
We previously described that the 95D cells had remarkably lower
4.1N/EPB41L1 expression than the 95C cells (Yang et al., 2016). Unexpectedly, although the methylation level of the 95D promoter
was higher than that of 95C, there was no significant difference
between
the
two
homologous
NSCLC
subclones
95C/95D
(Figure 5A). Over half of all protein-encoding genes are
regulated by miRNAs (Turchinovich et al., 2012). Aberrantly
high-expressed oncomiRNA silencing cancer suppressing genes
are frequently found in NSCLC. Therefore, we investigated the
potential miRNAs regulating 4.1N/EPB41L1 expression. The
miRNA-target gene database TargetScan (miRBase:www.mirbase. org) was used to predict the potential miRNAs regulating 4.1N/
EPB41L1, and miR-454-3p was suggested as a potential miRNA
(Figure 5B). qPCR results showed that the miR-454-3p was
expressed significantly lower in the 95D cells than in 95C cells
(Figure 5C). After overexpressing the miR-454-3p in A549 cells
(Figure 5D), the expression level of protein 4.1N/EPB41L1 was
downregulated (Figure 5E). Abnormally High-Expressed miR-454-3p
Decreases 4.1N/EPB41L1 in NSCLC ec eases
/
SC C
TCGA data were used to extract RNA transcript levels of the miR-
454-3p. We found that the miR-454-3p was significantly higher in
the LUAD (Figure 6A) and LUSC (Figure 6C) tissues in
comparison to the adjacent tissues, whereas the 4.1N/EPB41L1
mRNA was significantly lower in LUAD (Figure 6B) and LUSC
(Figure 6D) tissues than the corresponding adjacent tissues. Then, we used qPCR to measure the miR-454-3p and 4.1N/
EPB41L1 mRNA expressions in 37 NSCLC tissues and 31 tumor-
adjacent tissues. We found that the expression patterns are
consistent with TCGA data (Figures 6E,F). Moreover, the
association between miR-454-3p and 4.1N/EPB41L1 mRNA
was validated by Spearman’s coefficient analysis. The MiR-
454-3p
expression
level
showed
a
significantly
negative
correlation with 4.1N/EPB41L1 mRNA (r = −0.63, p < 0.0001,
Figure 6G). Expression patterns of miR-454-3p and 4.1N/
EPB41L1 mRNA showed that abnormally high expression of
miR-454-3p negatively regulates 4.1N/EPB41L1 in NSCLC. To further examine if 4.1N/EPB41L1 was a target gene of miR-
454-3p, the dual-luciferase activity assay was applied in A549 and
H460 cells. The predicted binding sites of miR-454-3p and 4.1N/
EPB41L1 mRNA and mutant sequences containing four mutated
nucleotides are shown in Figure 5F. MiR-454-3p significantly
suppressed the luciferase activity, and this suppressive effect was FIGURE 2 | 4.1N/EPB41L1 was restored by methyltransferase inhibitor 5-Aza-CdR in NSCLC cells. (A–C) qPCR was performed to evaluate expressions of 4.1N/
EPB41L1 mRNA in 95C, H460, and A549 cells after 0 μM or 1 μM 5-Aza-CdR treatments for 48 h (D–F) 95C, H460, and A549 cells were treated with 0, 1 or 10 μM 5-
Aza-CdR for 48 h, and then the protein was detected by Western blot. The data were presented as the mean ± SD, *p < 0.05. FIGURE 2 | 4.1N/EPB41L1 was restored by methyltransferase inhibitor 5-Aza-CdR in NSCLC cells. (A–C) qPCR was performed to evaluate expressions of 4.1N/
EPB41L1 mRNA in 95C, H460, and A549 cells after 0 μM or 1 μM 5-Aza-CdR treatments for 48 h (D–F) 95C, H460, and A549 cells were treated with 0, 1 or 10 μM 5-
Aza-CdR for 48 h, and then the protein was detected by Western blot. The data were presented as the mean ± SD, *p < 0.05. June 2022 | Volume 13 | Article 805960 5 Frontiers in Genetics | www.frontiersin.org Frontiers in Genetics | www.frontiersin.org 4.1N/EPB41L1 is Epigenetic Silenced Yang et al. Yang et al. Abnormally High-Expressed miR-454-3p
Decreases 4.1N/EPB41L1 in NSCLC FIGURE 3 | Methylation levels and prognosis of the 4.1N/EPB41L1 promoter in patients with LUAD. (A) Kaplan–Meier curves for cg13399773-4.1N showing
survival in lower (beta <0.611, shown in blue) and higher methylation groups (beta > 0.611, shown in red) dichotomized by the maxstat method. (C) Kaplan–Meier curves
for cg20993403-4.1N showing survival in lower (beta <0.59, shown in blue) and higher methylation groups (beta > 0.59, shown in red) dichotomized by the maxstat
method. (B,D) Violin plots showing the methylation levels of cg13399773-4.1N and cg20993403-4.1N among stage I–IV LUAD patients. Median methylation levels
(show by a thick black line) and interquartile range were summarized by the boxplot within each violin plot. HR: hazard ratio; LR: log-likelihood ratio; LUAD: lung
adenocarcinoma. Methylation levels were represented as beta values. Any beta value equal to or greater than 0.6 was considered fully methylated. Any beta value equal
to or less than 0.2 was considered to be fully unmethylated. Beta values between 0.2 and 0.6 were considered to be partially methylated. FIGURE 3 | Methylation levels and prognosis of the 4.1N/EPB41L1 promoter in patients with LUAD. (A) Kaplan–Meier curves for cg13399773-4.1N showing
survival in lower (beta <0.611, shown in blue) and higher methylation groups (beta > 0.611, shown in red) dichotomized by the maxstat method. (C) Kaplan–Meier curves
for cg20993403-4.1N showing survival in lower (beta <0.59, shown in blue) and higher methylation groups (beta > 0.59, shown in red) dichotomized by the maxstat
method. (B,D) Violin plots showing the methylation levels of cg13399773-4.1N and cg20993403-4.1N among stage I–IV LUAD patients. Median methylation levels
(show by a thick black line) and interquartile range were summarized by the boxplot within each violin plot. HR: hazard ratio; LR: log-likelihood ratio; LUAD: lung
adenocarcinoma. Methylation levels were represented as beta values. Any beta value equal to or greater than 0.6 was considered fully methylated. Any beta value equal
to or less than 0.2 was considered to be fully unmethylated. Beta values between 0.2 and 0.6 were considered to be partially methylated. DISCUSSIONS et al., 2005; Zhang et al., 2012), breast cancer (Heller et al., 2007), renal
clear cell carcinoma (Yamada et al., 2006), and prostate cancer (Schulz
et al., 2007; Schulz et al., 2010). Thus, we decided to explore the
underlying epigenetic disruptions of 4.1N/EPB41L1 deficiency in
NSCLC. Similarly,
we
found
that
high
4.1N/EPB41L1
gene
methylation was prevalent in LUAD and LUSC. 4.1B/EPB41L3
gene methylation is a potential indicator for poor prognosis in
NSCLC patients, especially in LUAD patients (Kikuchi et al., 2005). We found that a higher methylation level of 4.1N/EPB41L1 gene CpG
sites (cg13399773 and cg20993403) was potential predictive markers of
shorter overall survival in LUAD patients. It is acceptable that patients
with higher promoter methylation within the 4.1N/EPB41L1 gene have
a shorter overall survival time because the methylation inhibits tumor
suppressor 4.1N/EPB41L1 expression at the transcriptional level. However, it is a different matter for LUSC, which remains to be DNA methylation in the promoter region results in gene repression,
which is one of the most well-defined epigenetic hallmarks. We
previously
revealed
that
downregulated
4.1N/EPB41L1
exerts
antitumor effects by activating the classical Wnt pathway and
C-MYC expression in NSCLC (Wang et al., 2016; Yang et al.,
2016; Yang et al., 2021). The Wnt pathway disruption driven by
methylation of promoter regions plays a key driving role in the high
CpG island methylated phenotype LUAD subtype. This subtype is also
significantly correlated with the overexpressed MYC gene (Cancer
Genome Atlas Research, 2014; Duruisseaux and Esteller, 2018). 4.1N/
EPB41L1 has many similar biological characteristics to its homologous
4.1B/EPB41L3. High promoter methylation of the 4.1B/EPB41L3
gene–induced gene-silencing frequently occurs in NSCLC (Kikuchi June 2022 | Volume 13 | Article 805960 Frontiers in Genetics | www.frontiersin.org 6 4.1N/EPB41L1 is Epigenetic Silenced Yang et al. Yang et al. FIGURE 4 | Methylation levels and prognosis of the 4.1N/EPB41L1 promoter in patients with LUSC. (A) Kaplan–Meier curves for cg13399773-4.1N showing
survival in lower (beta <0.675, shown in blue) and higher methylation groups (beta > 0.675, shown in red) dichotomized by the maxstat method. (C) Kaplan–Meier curves
for cg07030373-4.1N showing survival in lower (beta <0.796, shown in blue) and higher methylation groups (beta > 0.796, shown in red) dichotomized by the maxstat
method. (E) Kaplan–Meier curves for cg20993403-4.1N showing survival in lower (beta <0.611, shown in blue) and higher methylation groups (beta > 0.611,
shown in red) dichotomized by the maxstat method. DISCUSSIONS (B,D,F) Violin plots showing the methylation levels of cg13399773-4.1N, cg07030373-4.1N, and cg20993403-
4.1N among stage I–IV LUAD patients. Median methylation levels (show by a thick black line) and interquartile range were summarized by the boxplot within each violin
plot. HR: hazard ratio; LR: log-likelihood ratio; LUSC: lung squamous cell carcinoma. Methylation levels were represented as beta values. Any beta value equal to or
greater than 0.6 was considered fully methylated. Any beta value equal to or less than 0.2 was considered to be fully unmethylated. Beta values between 0.2 and 0.6
were considered to be partially methylated. FIGURE 4 | Methylation levels and prognosis of the 4.1N/EPB41L1 promoter in patients with LUSC. (A) Kaplan–Meier curves for cg13399773-4.1N showing
survival in lower (beta <0.675, shown in blue) and higher methylation groups (beta > 0.675, shown in red) dichotomized by the maxstat method. (C) Kaplan–Meier curves
for cg07030373-4.1N showing survival in lower (beta <0.796, shown in blue) and higher methylation groups (beta > 0.796, shown in red) dichotomized by the maxstat
method. (E) Kaplan–Meier curves for cg20993403-4.1N showing survival in lower (beta <0.611, shown in blue) and higher methylation groups (beta > 0.611,
shown in red) dichotomized by the maxstat method. (B,D,F) Violin plots showing the methylation levels of cg13399773-4.1N, cg07030373-4.1N, and cg20993403-
4.1N among stage I–IV LUAD patients. Median methylation levels (show by a thick black line) and interquartile range were summarized by the boxplot within each violin
plot. HR: hazard ratio; LR: log-likelihood ratio; LUSC: lung squamous cell carcinoma. Methylation levels were represented as beta values. Any beta value equal to or
greater than 0.6 was considered fully methylated. Any beta value equal to or less than 0.2 was considered to be fully unmethylated. Beta values between 0.2 and 0.6
were considered to be partially methylated. elucidated in the future. The 5-year survival rate is less than 15% for
NSCLC patients, but the rate can increase to 63% with the early stage of
initial diagnosis (van Rens et al., 2000; Geng et al., 2017), thus
demonstrating the value of the early diagnosis of NSCLC. 4.1B/
EPB41L3 gene promoter methylation is regarded as an early event
in renal clear cell carcinoma (Yamada et al., 2006). In this study, high
4.1N/EPB41L1 gene methylation was also observed to be a relatively
early event in LUAD and LUSC patients, indicating its valuable role in
tumorigenesis and potential as an early detection marker. DISCUSSIONS qPCR results of our sample set and the TCGA database
together showed that the miR-454-3p was upregulated in the
NSCLC tumor tissue, which contrasted with the results from
another independent study (Jin et al., 2019). Interestingly, tumor
expression of miR-454-5p in NSCLC is reported as upregulated
(Zhu et al., 2016), but another report suggests its expression to be
downregulated (Liu et al., 2018). It is not rare when various
clinical specimens are evaluated; the expression levels of some
miRNAs are different. Because of the complexity of NSCLC Frontiers in Genetics | www.frontiersin.org June 2022 | Volume 13 | Article 805960 7 4.1N/EPB41L1 is Epigenetic Silenced Yang et al. Yang et al. FIGURE 5 | MiR-454-3p directly bound to 3′UTR of 4.1N/EPB41L1 mRNA. (A) There was no significant difference in methylation levels of the 4.1N/EPB41L1 gene
promoter between the two homologous NSCLC subclones 95C and 95D. (B) MiR-454-3p was a predicted binding partner of 4.1N/EPB41L1. (C,D) After transfecting
A549 cells with miR-454-3p or negative control mimics, the expression levels of miR-454-3p and protein 4.1N/EPB41L1 were detected using qPCR and Western blot,
respectively. (E) miR-454-3p expression was significantly lower in 95C cells than 95D cells. (F) Schematic representation of the predicted binding sites of miR-454-
3p and 4.1N/EPB41L1 mRNA and the mutated sequences in potential binding sites. (G,H) A549 or H460 cells were co-transfected with miR-454-3p or control mimics
and wild-type (psiCHECK2-4.1N-wt) or mutant-type (psiCHECK2-4.1N-mut) plasmids. 48 h later, dual-luciferase activity was measured. The data were presented as the
mean ± SD. *p < 0.05 GADPH and U6 were used as loading control of 4.1N/EPB41L1 mRNA and miR-454-3p, respectively. FIGURE 5 | MiR-454-3p directly bound to 3′UTR of 4.1N/EPB41L1 mRNA. (A) There was no significant difference in methylation levels of the 4.1N/EPB41L1 gene
promoter between the two homologous NSCLC subclones 95C and 95D. (B) MiR-454-3p was a predicted binding partner of 4.1N/EPB41L1. (C,D) After transfecting
A549 cells with miR-454-3p or negative control mimics, the expression levels of miR-454-3p and protein 4.1N/EPB41L1 were detected using qPCR and Western blot,
respectively. (E) miR-454-3p expression was significantly lower in 95C cells than 95D cells. (F) Schematic representation of the predicted binding sites of miR-454-
3p and 4.1N/EPB41L1 mRNA and the mutated sequences in potential binding sites. (G,H) A549 or H460 cells were co-transfected with miR-454-3p or control mimics
and wild-type (psiCHECK2-4.1N-wt) or mutant-type (psiCHECK2-4.1N-mut) plasmids. 48 h later, dual-luciferase activity was measured. DISCUSSIONS The data were presented as the
mean ± SD. *p < 0.05 GADPH and U6 were used as loading control of 4.1N/EPB41L1 mRNA and miR-454-3p, respectively. breast cancer (Ren et al., 2019; Wang et al., 2020), and colon
cancer (Li et al., 2018). Moreover, an integrative bioinformatics
analysis indicates that miR-454-3p is a biomarker for diagnosing
some cancers, including the LUAD (Botling et al., 2013), which
needs further confirmation studies in the future. development, for these miRNAs, the specimen integration of
different NSCLC stages and histological types could impede their
expressions together (Zhong et al., 2021). In NSCLC, our study
showed aberrantly high-expressed miR-454-3p directly bound to
4.1N/EPB41L1 mRNA 3′UTR and led to the depression of tumor
suppressor
4.1N/EPB41L1
at
the
posttranscriptional
level,
uncovering the known miRNAs regulating 4.1N/EPB41L1. It
has
been
reported
that
the
miR-454-3p
also
acts
as
oncomiRNA in oral squamous cell carcinoma (Shi et al.,
2021), cervical cancer (Guo et al., 2018; Shukla et al., 2019;
Song et al., 2019; Li et al., 2021), liver cancer (Li et al., 2019), Epigenetic alterations have been demonstrated to be highly
orchestrated from the initiation step to therapy resistance step in
lung cancer (Quintanal-Villalonga and Molina-Pinelo, 2019). Although many
research
studies
have revealed
the vital
anticancer roles of 4.1 family members, the knowledge of the
underlying epigenetic mechanism behind their loss in cancers is June 2022 | Volume 13 | Article 805960 Frontiers in Genetics | www.frontiersin.org 8 4.1N/EPB41L1 is Epigenetic Silenced Yang et al. Yang et al. | Validation of expression patterns of miR-454-3p and 4.1N/EPB41L1 mRNA in NSCLC. Relative expressions of miR-454-3p a
UAD. (A,B) and LUSC (C,D) were analyzed using the data from the TCGA dataset. (E-F) Relative expression of miR-454-3p and 4
d adjacent tissues was measured by qPCR. GADPH and U6 were used as loading control of 4.1N/EPB41L1 mRNA and miR-4
between miR-454-3p and 4.1N/EPB41L1 mRNA was determined using Spearman’s coefficient analysis. The data were presented
and ***p < 0.001. FIGURE 6 | Validation of expression patterns of miR-454-3p and 4.1N/EPB41L1 mRNA in NSCLC. Relative expressions of miR-454-3p and 4.1N/EPB41L1
mRNA in LUAD. (A,B) and LUSC (C,D) were analyzed using the data from the TCGA dataset. (E-F) Relative expression of miR-454-3p and 4.1N/EPB41L1 mRNA in
NSCLC and adjacent tissues was measured by qPCR. GADPH and U6 were used as loading control of 4.1N/EPB41L1 mRNA and miR-454-3p, respectively. REFERENCES Gao, F., Xia, Y., Wang, J., Lin, Z., Ou, Y., Liu, X., et al. (2014). Integrated Analyses of
DNA Methylation and Hydroxymethylation Reveal Tumor Suppressive Roles
of ECM1, ATF5, and EOMESin Human Hepatocellular Carcinoma. Genome
Biol. 15 (12), 533. doi:10.1186/s13059-014-0533-9 Arechederra, M., Daian, F., Yim, A., Bazai, S. K., Richelme, S., Dono, R., et al. (2018). Hypermethylation of Gene Body CpG Islands Predicts High Dosage of
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Preterm Neonates. BMC Genomics 2615, 716. doi:10.1186/1471-2164-15-716 Botling, J., Edlund, K., Lohr, M., Hellwig, B., Holmberg, L., Lambe, M., et al. (2013). Biomarker Discovery in Non-small Cell Lung Cancer: Integrating Gene
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Diagnosis, Treatment and Follow-Up. Ann. Oncol. 21 (Suppl. 5), v116–v119. doi:10.1093/annonc/mdq189 Heller, G., Geradts, J., Ziegler, B., Newsham, I., Filipits, M., Markis-Ritzinger, E.-
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Frequently in Breast Cancer. Breast Cancer Res. Treat. 103 (3), 283–291. doi:10. 1007/s10549-006-9377-7 Du, X., Zhang, J., Wang, J., Lin, X., and Ding, F. (2018). Role of miRNA in Lung
Cancer-Potential Biomarkers and Therapies. Cpd 23 (39), 5997–6010. doi:10. AUTHOR CONTRIBUTIONS QY, LZ, HL, and JL contributed the central idea, performed the
experiments, analyzed the data, and wrote the initial draft of the
manuscript. The remaining authors discussed the results and
finalized this manuscript. All authors read and approved the final
manuscript. FUNDING This study was funded by the National Natural Science
Foundation of China (Grant Nos 81770107 and 82003286);
the Natural Science Foundation of Hunan Province (Grant
No. 2020JJ4560); Scientific Research Foundation of Hunan
Provincial
Education
Department
(Grant
No. 20B528);
Guidance Science and Technology Program of Shaoyang City
(Grant No. 2019ZD07); the Fellowship of China Postdoctoral
Science
Foundation
(Grant
Nos
2020M672474
and
2021T140195); and the Changsha Municipal Natural Science
Foundation
(Grant
Number
kq20140421);
Postgraduate
Innovation and Research Project of Central South University
(Grant No. 2021zzts0569). DATA AVAILABILITY STATEMENT Publicly available datasets were analyzed in this study. These data
can be found here: To investigate the 4.1N/EPB41L1 gene
methylation and its relation to NSCLC patient prognosis, TCGA
methylation data (https://cancergenome.nih.gov/) of the array
features 450k CpG sites covering the 4.1N gene promoter region
(TSS200 and TSS1500) were analyzed. The MethSurv tool (https://
biit.cs.ut.ee/methsurv/) was used to perform the assessment of
methylation-based analysis for the 4.1N/EPB41L1 gene in lung
adenocarcinoma (LUAD) and lung squamous cell carcinoma
(LUSC). TCGA data for LUAD and LUSC were used to extract
RNA transcript levels of the miR-454-3p. DISCUSSIONS (G)
Correlation between miR-454-3p and 4.1N/EPB41L1 mRNA was determined using Spearman’s coefficient analysis. The data were presented as the mean ± SD. *p < 0.05 and ***p < 0.001. FIGURE 6 | Validation of expression patterns of miR-454-3p and 4.1N/EPB41L1 mRNA in NSCLC. Relative expressions of miR-454-3p and 4.1N/EPB41L1
mRNA in LUAD. (A,B) and LUSC (C,D) were analyzed using the data from the TCGA dataset. (E-F) Relative expression of miR-454-3p and 4.1N/EPB41L1 mRNA in
NSCLC and adjacent tissues was measured by qPCR. GADPH and U6 were used as loading control of 4.1N/EPB41L1 mRNA and miR-454-3p, respectively. (G)
Correlation between miR-454-3p and 4.1N/EPB41L1 mRNA was determined using Spearman’s coefficient analysis. The data were presented as the mean ± SD. *p < 0.05 and ***p < 0.001. June 2022 | Volume 13 | Article 805960 Frontiers in Genetics | www.frontiersin.org 9 4.1N/EPB41L1 is Epigenetic Silenced Yang et al. Hospital, Central South University. The patients/participants
provided their written informed consent to participate in this
study. nearly empty. This study showed that promoter methylation and
miR-454-3p were implicated in the expression deregulation of the
4.1N/EPB41L1 at transcriptional and posttranscriptional levels,
respectively. The epigenetic abnormalities are reversible; the
application of upregulating 4.1N/EPB41L1 by targeting DNA
methylation and miR-454-3p may represent a promising
therapy for NSCLC treatment. ETHICS STATEMENT The studies involving human participants were reviewed and
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access article distributed under the terms of the Creative Commons Attribution
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provided the original author(s) and the copyright owner(s) are credited and that the
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Genomic and transcriptomic analysis of pituitary adenomas reveals the impacts of copy number variations on gene expression and clinical prognosis among prolactin-secreting subtype
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Yiyuan Chen
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ABSTRACT Pituitary adenomas (PAs) are slow growing and benign primary intracranial tumors that often cause occupying
effects or endocrine symptoms. PAs can be classified into various subtypes according to hormone secretion. Although widespread transcriptional alterations that cause aberrant hormone secretion have been characterized,
the impact of genomic variations on transcriptional alterations is unclear due to the rare occurrence of single-
nucleotide variations in PA. In this study, we performed whole-genome sequencing (WGS) on 76 PA samples
across three clinical subtypes (PRL-PAs; GH-PAs, and NFPAs); transcriptome sequencing (RNA-seq) of 54 samples
across these subtypes was also conducted. Nine normal pituitary tissues were used as controls. Common and
subtype-specific transcriptional alterations in PAs were identified. Strikingly, widespread genomic copy number
amplifications were discovered for PRL-PAs, which are causally involved in transcriptomic changes in this subtype. Moreover, we found that the high copy number variations (CNVs) in PRL-PA cause increased prolactin production,
drug resistance and proliferative capacity, potentially through key genes with copy number amplification and
transcriptional activation, such as BCAT1. This study provides insight into how genomic CNVs affect the
transcriptome and clinical outcomes of PRL-PA and sheds light on the development of potential therapeutics for
aberrantly activated targets. AGING 2021, Vol. 13, No. 1 www.aging-us.com AGING 2021, Vol. 13, No. 1 p
Genomic and transcriptomic analysis of pituitary adenomas reveals
the impacts of copy number variations on gene expression and clinical
prognosis among prolactin-secreting subtype Yiyuan Chen1, Hua Gao1, Weiyan Xie1, Jing Guo1, Qiuyue Fang1, Peng Zhao2, Chunhui Liu2, Haibo
Zhu2, Zhuang Wang3, Jichao Wang4, Songbai Gui2,5,6, Yazhuo Zhang1,2,5,6, Chuzhong Li1,2,5,6 1Department of Cell Biology, Beijing Neurosurgical Institute, Capital Medical University, Beijing 100070, China
2Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing 100070, China
3Department of Neurosurgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
4Department of Neurosurgery, People's Hospital of Xinjiang Uygur Autonomous Region, Xinjiang 830001, China
5China National Clinical Research Center for Neurological Diseases, Beijing 100070, China
6Brain Tumor Center, Beijing Institute for Brain Disorders, Beijing 100070, China Correspondence to: Chuzhong Li; email: lichuzhong@ccmu.edu.cn
Keywords: pituitary adenoma, WGS, CNV, bromocriptine resistance
Received: August 8, 2020
Accepted: September 29, 2020 Correspondence to: Chuzhong Li; email: lichuzhong@ccmu.edu.cn
Keywords: pituitary adenoma, WGS, CNV, bromocriptine resistance
Received: August 8, 2020
Accepted: September 29, 2020
Published: December 19, 2020
Copyright: © 2020 Chen et al. This is an open access article distributed under the terms of the Creative Commons Attribution
License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original
author and source are credited. Copyright: © 2020 Chen et al. This is an open access article distributed under the terms of the Creative Commons Attribution
License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original
author and source are credited. Copyright: © 2020 Chen et al. This is an open access article distributed under the terms of the Creative Commons Attribution
License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original
author and source are credited. INTRODUCTION status, and prolactin-secreting PAs (PRL-PAs, 32%-51%)
and growth hormone-secreting PAs (GH-PAs, 9%-11%)
are the two most common subtypes accounting for a large
percentage of functional PAs. Adrenocorticotropin-
secreting PAs (ACTH-PAs, 3%-6%) and thyrotropin-
secreting PAs (TSH-PAs, <1%) can cause obvious
clinical symptoms, but their incidence is lower than that
of PRL-PAs and GH-PAs [2, 5, 6]. Most gonadotropin Pituitary adenomas (PAs) are primary intracranial tumors
that occur in 0.1% of adults [1, 2]. PAs can be classified
according to the presence of abnormal hormone secretion
(functional PAs, 36%-54%; clinical nonfunctional PAs,
NFPAs, 46%-64%) [3, 4]. Functional PAs can be further
divided into subtypes according to hormone secretion AGING 1276 www.aging-us.com adenomas (GON-PAs) and pluri-hormonal PAs are
NFPAs. The majority of pituitary tumors are benign and
exhibit arrested cell cycle as well as aberrant growth
factor signaling [7, 8]. specimens clustered according to their clinical groups,
suggesting widespread transcriptomic alterations in PAs
and across PA subtypes and that transcriptional
signatures can be used for molecular classification of PA
subtypes. The similarity heatmap suggests that NFPAs
exhibit
more
dramatic
transcriptomic
alterations
(Figure 1A). Consistently, the number of DEGs between
NFPAs and other groups (3800~4200) was significantly
larger than the number of DEGs between other
comparison groups (Supplementary Figures 1, 2). This
result suggests that despite a lack of abnormal hormone
secretion, NFPAs are characterized by more widespread
gene expression alterations. Transcription level alterations have been linked to
abnormal hormone secretion in different subtypes of
PAs [9]. In ACTH-PAs, the proopiomelanocortin
(POMC), T-Box transcription factor 19 (TBX19, TPIT)
and epidermal growth factor receptor (EGFR) genes are
reportedly upregulated [10]; aberrant splicing of
estrogen-related receptor gamma (ESRRG) leads to
stronger binding to POU class 1 homeobox 1 (POU1F1,
PIT-1)
and
excessive
prolactin
secretion
[11]. Transcriptomic and epigenomic approaches have been
employed to illustrate divergent patterns of gene
expression in several subtypes of PAs [12]. However,
the global changes in gene expression in PAs are still
under investigation due to the lack of normal pituitary
tissue as a control. A total of 448 common DEGs were found to be shared
across PA subtypes compared to normal pituitary tissue
(Figure 1B). These genes are enriched with neuronal
pathways, including neural active ligand-receptor
binding and axon guidance (Figure 1C). INTRODUCTION The common
DEGs are also enriched in five growth factor signaling
pathways (Wnt, TGF-β, PI3K-AKT, Hippo, and STAT3-
JAK), all of which have been linked to PA pathogenesis
or therapy [17–21]. These genes commonly changed
across PA subtypes are also involved in cytokine
production pathways (Figure 1C), consistent with
decreased CD8+ T cell infiltration in both functional PAs
and NFPAs (Figure 1D, 1E) and suggesting suppression
of the immune response as a common etiology of PAs. Genomic analyses have identified genes with somatic
single-nucleotide variations (SNVs) in PAs, e.g.,
Ubiquitin-specific peptidase 8 (USP8) in ACTH-PAs, G
protein αs (GNAS) in GH-PAs and splicing factor 3b
subunit 1 (SF3B1) in PRL-PAs [13–16]. However, these
genomic studies suggested that PAs are associated with
low mutation burdens; in addition, the frequencies of the
three recurrent mutations were quite low, indicating a
limited impact of SNVs on widespread gene expression
alterations in PAs [12]. The impact of other genomic
alterations on transcriptomic changes, such as copy
number variations, remains to be investigated. Transcriptional signatures reveal altered pathways
across PA subtypes To identify transcriptional signatures associated with
each PA subtype, 54 transcriptomic datasets were used to
construct a weighted gene coexpression network [22]
consisting of 62 modules with a size of 30~1500 genes
(Figure 2A). Analysis of module trait association
revealed modular alterations of gene expression in each
subtype (Figure 2B and Supplementary Figure 3). The
two abnormal hormone-secretion subtypes share some
coexpression modules (e.g., MEturquoise, MEgreen and
MEdarkgreen), and the module MEturquoise genes was
enriched with pathways related to protein synthesis
(Figure 2C, 2D), indicating that abnormal macromolecule
biosynthesis may be the basis of aberrant hormone
production and secretion. Several coexpression modules
were significantly associated with the nonfunctional PA
subtype, e.g., the Module purple showed enrichment of
genes in the insulin signaling pathway (Figure 2E). As
patients with hormone-abnormal PA usually develop
insulin resistance and glucose abnormalities [23],
activated insulin signaling in NFPAs might antagonize
metabolic disorders, thus preventing excessive hormone
secretion. The high expression of genes in subtype-
correlated modules was confirmed by the heatmaps
(Figure 2F). In this study, we performed whole-genome sequencing
(WGS) and transcriptomic sequencing (RNA-seq) on
PRL-PAs, GH-PAs and NFPAs to identify subtype-
specific genomic and transcriptomic alterations. Normal pituitary tissues were also used to identify
common gene expression abnormalities in PAs. Common gene expression alterations were detected in
PAs, including genes in neuronal pathways and growth
pathways. Widespread and unrecognized genomic
copy number amplifications were identified in PRL-
PAs, contributing to specific transcriptional activation
in numerous genes and worse clinical outcomes of
PRL-PA patients. Genomic copy number amplifications causally
involved in gene transcriptional activation in PRL-PAs PAs, were identified with a low frequency of
occurrence (Supplementary Material 1). Strikingly, we
found
widespread
and
recurrent
copy
number
variations, especially amplifications, in PRL-PAs. The
high-CNV feature occurred in nearly half of the PRL-
PA cases but rarely in other PA subtypes (Figure 3A). Specific genomic copy number amplification was not
mentioned in a previous exome sequencing analysis of
PAs, possibly because of the low number of PRL-PA
patients involved. Next, we attempted to explore genomic alterations
underlying the observed transcriptomic changes in PA
patients. WGS data from 76 PA samples and paired
blood samples were analyzed. We noticed a similar low
mutation burden in these PA samples, consistent with
previous observations. Some previously reported SNVs
associated with PAs, such as GNAS and 1/11 in GH- Figure 1. The transcriptional landscape of PAs. (A) Correlation heatmap of transcriptomic similarity among 21 PRL-PAs, 11 GH-PAs, 23
NFPAs and 9 normal pituitary tissues (Normal). Pituitary adenoma subtype is indicated by the color bar above the heatmap. (B) Venn diagram
showing the intersection of DEGs among three subtypes of PAs vs. Normal. DEGs were identified by the R package DESeq2 under the cutoff
of adjusted P value < 0.05. (C) KEGG pathway enrichment analysis of 448 overlapping DEGs, the dot plot shows pathways with an adjusted P
value < 0.05. (D) The infiltration of eight subtypes of immune cell populations and two endothelial cell types in PA samples and normal
pituitary tissues was evaluated using the expression levels of cell type specific markers using the MCP-counter [38]. The abundances of each
cell types were normalized by z transformation across samples. (E) Boxplots of z score from Figure 1D showing the reduced infiltration of
CD8+ T cells was reduced across PA subtypes compared to normal pituitary tissues. Figure 1. The transcriptional landscape of PAs. (A) Correlation heatmap of transcriptomic similarity among 21 PRL-PAs, 11 GH-PAs, 23
NFPAs and 9 normal pituitary tissues (Normal). Pituitary adenoma subtype is indicated by the color bar above the heatmap. (B) Venn diagram
showing the intersection of DEGs among three subtypes of PAs vs. Normal. DEGs were identified by the R package DESeq2 under the cutoff
of adjusted P value < 0.05. (C) KEGG pathway enrichment analysis of 448 overlapping DEGs, the dot plot shows pathways with an adjusted P
value < 0.05. Transcriptional landscape of pituitary tumors We performed transcriptome sequencing (RNA-seq) on
54 PA samples (21 PRL-PAs, 11 GH-PAs, and 23
NFPAs) and 9 normal pituitary tissues (Supplementary
Table 1). Hierarchical clustering showed that these AGING 1277 www.aging-us.com Genomic copy number amplifications causally
involved in gene transcriptional activation in PRL-PAs (D) The infiltration of eight subtypes of immune cell populations and two endothelial cell types in PA samples and normal
pituitary tissues was evaluated using the expression levels of cell type specific markers using the MCP-counter [38]. The abundances of each
cell types were normalized by z transformation across samples. (E) Boxplots of z score from Figure 1D showing the reduced infiltration of
CD8+ T cells was reduced across PA subtypes compared to normal pituitary tissues. AGING 1278 www.aging-us.com Figure 2. Weighted gene correlation network analysis (WGCNA) of transcriptome data reveals subtype specific coexpression
modules. (A) WGCNA cluster dendrogram groups genes into distinct gene coexpression modules defined by dendrogram branch cutting
(B) Modules-subtype correlation heatmap of three PA subtypes and normal tissues. The cells in the heatmap were colored by the correlation
between eigengene expression and each sample group, the correlation coefficients and P values were indicated in each cell. (C–E) KEGG
pathway enrichment analysis of genes in the modules significantly associated with different PA subtypes. Adjusted P value < 0.05
(F) Expression profiles of genes in key modules associated with PA subtypes. Figure 2. Weighted gene correlation network analysis (WGCNA) of transcriptome data reveals subtype specific coexpression
modules. (A) WGCNA cluster dendrogram groups genes into distinct gene coexpression modules defined by dendrogram branch cutting. (B) Modules-subtype correlation heatmap of three PA subtypes and normal tissues. The cells in the heatmap were colored by the correlation
between eigengene expression and each sample group, the correlation coefficients and P values were indicated in each cell. (C–E) KEGG
pathway enrichment analysis of genes in the modules significantly associated with different PA subtypes. Adjusted P value < 0.05. (F) Expression profiles of genes in key modules associated with PA subtypes. AGING AGING 1279 www.aging-us.com Table 1. Clinico-pathological characteristics of high- and low- CNV PRL-PAs. Genomic copy number amplifications causally
involved in gene transcriptional activation in PRL-PAs Sample
Volume1
Invasive/non-Invasive
CNV2
Drug resistance3
Ki-67 IHC4
P906
Large
Invasive
High
Resistance
8%
P918
Large
non-Invasive
High
NA
<1%
P1133
Large
non-Invasive
High
NA
2%
P1154
Giant
Invasive
High
NA
>3%
P1037
Giant
Invasive
High
Resistance
3%
P1408
Giant
non-Invasive
High
Resistance
<1%
P1100
Large
non-Invasive
High
Resistance
<1%
P1694
Large
Invasive
High
NA
5-8%
P1711
Large
non-Invasive
High
NA
NA
P1712
Giant
Invasive
High
Resistance
6%
P1736
Giant
Invasive
High
Resistance
8%
P1809
Large
Invasive
High
Resistance
7%
P25502
Large
Invasive
High
Sensitive
3-5%
P28607
Large
Invasive
High
Resistance
1-2%
P2890
Large
non-Invasive
High
NA
2-3%
P1824
Giant
Invasive
High
Sensitive
5-10%
P25505
Large
Invasive
High
Resistance
<1%
P1821
Large
non-Invasive
High
Resistance
NA
P1070
Large
non-Invasive
Low
Sensitive
2%
P961
Giant
Invasive
Low
NA
<1%
P1483
Giant
Invasive
Low
Sensitive
<1%
P1587
Large
Invasive
Low
NA
NA
P_N6_30
Large
non-Invasive
Low
Sensitive
<1%
P1199
Large
Invasive
Low
Resistance
<1%
P1144
Large
Invasive
Low
Sensitive
<1%
P1825
Giant
Invasive
Low
NA
1%
P640829
Large
Invasive
Low
NA
NA
P29115
Large
Invasive
Low
Sensitive
2-5%
1 Tumor classification by volume, micro: <1 cm, large: 1-4 cm, giant: >4 cm. 2 According to the hierarchical clustering of CNV signatures. 3 Drug (BCT) resistance, NA: no drug therapy was administered preoperatively. 4 Ki-67 IHC staining, NA: not enough tissue. Table 1. Clinico-pathological characteristics of high- and low- CNV PRL-PAs. g
g
g
3 Drug (BCT) resistance, NA: no drug therapy was administered preoperatively. 4 Ki-67 IHC staining, NA: not enough tissue. PRL-PA specific upregulated genes was significantly
higher in the high-CNV group (Figure 3D), suggesting
a causal role of CNVs in transcriptional changes in
PRL-PA patients. The genes located in copy number amplification regions
significantly overlapped with the genes specifically
upregulated in PRL-PAs compared to other subtypes
(Figure 3B), implying a role of genomic CNV in
shaping transcriptomic alterations in PRL-PAs. The 498
overlapping
genes
were
enriched
in
pathways
downstream
of
mTOR signaling, including the
biosynthesis of amino acids, lysosome pathway,
ribosome and AMPK signaling (Figure 3C). Cyclin-
dependent kinase 6 (CDK6) was also transcriptionally
activated by copy number amplification, which suggests
the role of genomic copy number variation in the out-
of-control cell cycle and tumor progression of PAs. 1 Tumor classification by volume, micro: <1 cm, large: 1-4 cm, giant: >4 cm.
2 Genomic copy number amplifications cause high
prolactin production and activation of genes
downstream of the mTOR signaling pathway To investigate the influence of genomic copy number
amplification on the clinical outcomes of PRL-PA
patients, we used an immunohistochemistry (IHC)
staining approach to probe key genes under copy number
amplifications in PRL-PAs. The copy number-amplified
genes BCAT1 and MYC were more abundant in the
high-CNV group of patients (Figure 4A). To further
analyze the pathological impacts of genomic copy
number variation, we surveyed expression of prolactin in We further divided PRL-PA patients into high-CNV
and low-CNV groups according to the hierarchical
clustering of CNV signatures. Consistent with the
increase in copy number, the expression fold change of AGING 1280 www.aging-us.com gure 3. Copy number amplifications in PRL-PAs cause gene transcriptional activation. (A) The heatmap of CNV profiles acro
A subtypes. The sample subtype was indicated by color bar above the heatmap, samples were grouped according to the similarity of C
rofiles. The heatmap was colored by CNVs, red indicates gain of copy number and blue indicates loss of copy number. Frequent co
umber amplification was observed in PRL-PAs. (B) 498 genes with the copy number amplifications in PRL-PAs overlapped with u
gulated DEGs in the PRL-PAs compared to other subtypes. P value < 2.2e-16 by hypergeometric test. (C) KEGG pathway enrichme
nalysis of 498 up-regulated genes with both copy number amplifications and transcriptional up-regulation in PRL-PAs, the dot plot sho
athways with a P value < 0.05. (D) PRL-PAs samples were divided into two groups (high CNV and low CNV) according to the cluster Figure 3. Copy number amplifications in PRL-PAs cause gene transcriptional activation. (A) The heatmap of CNV profiles across
PA subtypes. The sample subtype was indicated by color bar above the heatmap, samples were grouped according to the similarity of CNV
profiles. The heatmap was colored by CNVs, red indicates gain of copy number and blue indicates loss of copy number. Frequent copy
number amplification was observed in PRL-PAs. (B) 498 genes with the copy number amplifications in PRL-PAs overlapped with up- Figure 3. Copy number amplifications in PRL-PAs cause gene transcriptional activation. (A) The heatmap of CNV profiles across
PA subtypes. The sample subtype was indicated by color bar above the heatmap, samples were grouped according to the similarity of CNV
profiles. The heatmap was colored by CNVs, red indicates gain of copy number and blue indicates loss of copy number. Frequent copy
number amplification was observed in PRL-PAs. Genomic copy number amplifications cause high
prolactin production and activation of genes
downstream of the mTOR signaling pathway (B) 498 genes with the copy number amplifications in PRL-PAs overlapped with up-
regulated DEGs in the PRL-PAs compared to other subtypes. P value < 2.2e-16 by hypergeometric test. (C) KEGG pathway enrichment
analysis of 498 up-regulated genes with both copy number amplifications and transcriptional up-regulation in PRL-PAs, the dot plot shows
pathways with a P value < 0.05. (D) PRL-PAs samples were divided into two groups (high CNV and low CNV) according to the clustering
results in (A). The boxplot shows the log2 expression fold-changes of PRL-PAs specific up-regulation genes relative to GH-PAs/NFPAs in the
high CNV group and low CNV group. The high CNV group exhibited transcriptional up-regulation (Median log2 fold change ~0.75) while the
low CNV group did not. AGING 1281 www.aging-us.com significant impact of genomic CNVs on the excessive
production of prolactin in PRL-PA patients. The
transcriptomic differences between the high- and low-
CNV groups also involved numerous ribosome genes
and branched-chain amino acid aminotransferase 1
(BCAT1), implicating activation of the mTOR signaling patients in the high- and low-CNV groups and found
prolactin expression to be 4-fold higher in the former
group (Figure 4B). ErbB signaling is a determinant of
prolactin production, and the downstream transcription
factor of ErbB signaling, MYC, also exhibited a 4-fold
increase in the high-CNV group. These results suggest a ure 4. Clinical relevance of genomic copy number variation in PRL-PAs. (A) Immunohistochemistry of BCAT1, MYC, Ki-67, PR
d PIT1 in high and low CNV PRL-PAs. ×100 magnification, the scale bar = 100 μm. (B) Prolactin expression levels (TPM) in PRL-PAs with hi
V, PRL-PAs with low CNV, GH-PAs and NFPAs. *P<0.05, **P<0.01, ***P<0.001 (C) Copy number and expression level (TPM) of BCAT1
ferent PA subtypes. Both copy number and expression level of BCAT1 were increased in PRL-PAs. (D) PRL-PA patients in the high CN
oup more frequently developed drug resistance. The yellow bar indicates the number of patients that are sensitive to BCT treatment, wh
e blue bar indicates number of patients that are insensitive to the same treatment. P value = 0.026, Chi-Square test. (E) High CNV group
L-PAs exhibits a higher degree of malignancy. The PA malignancy was defined by the number of positive Ki-67 foci in IHC. P value = 0.05
-Square test. Genomic copy number amplifications cause high
prolactin production and activation of genes
downstream of the mTOR signaling pathway (F) The Scatter plot shows positive correlation (spearman correlation coefficient: 0.47) of transcriptomic alterations in hig
V group relative to low CNV group (x-axis) and relapsed PAs relative to un-relapsed PAs (y-axis). Figure 4. Clinical relevance of genomic copy number variation in PRL-PAs. (A) Immunohistochemistry of BCAT1, MYC, Ki-67, PRL,
and PIT1 in high and low CNV PRL-PAs. ×100 magnification, the scale bar = 100 μm. (B) Prolactin expression levels (TPM) in PRL-PAs with high
CNV, PRL-PAs with low CNV, GH-PAs and NFPAs. *P<0.05, **P<0.01, ***P<0.001 (C) Copy number and expression level (TPM) of BCAT1 in
different PA subtypes. Both copy number and expression level of BCAT1 were increased in PRL-PAs. (D) PRL-PA patients in the high CNV
group more frequently developed drug resistance. The yellow bar indicates the number of patients that are sensitive to BCT treatment, while
the blue bar indicates number of patients that are insensitive to the same treatment. P value = 0.026, Chi-Square test. (E) High CNV group in
PRL-PAs exhibits a higher degree of malignancy. The PA malignancy was defined by the number of positive Ki-67 foci in IHC. P value = 0.059,
Chi-Square test. (F) The Scatter plot shows positive correlation (spearman correlation coefficient: 0.47) of transcriptomic alterations in high
CNV group relative to low CNV group (x-axis) and relapsed PAs relative to un-relapsed PAs (y-axis). Figure 4. Clinical relevance of genomic copy number variation in PRL-PAs. (A) Immunohistochem AGING 1282 www.aging-us.com pathway in the high-CNV group (Figure 4C). BCAT1
undergoes correlated copy number amplifications and
gene expression increments in PRL-PAs. Both BCAT1
and prolactin can activate the mTOR signaling pathway,
which leads to excessive ribosome biogenesis. alterations, as opposed to aberration of specific
hormonal pathways. We utilized WGCNA to generate coexpression networks
in PAs and identified subtype-specific modules. The
functional subtypes (GH-PAs and PRL-PAs) shared
modules enriched in growth hormone pathways; PRL-
PAs were also enriched in ribosome genes and the
mitochondrial
oxidative
phosphorylation
pathways,
suggesting a high demand of energy metabolism in the
PRL-PAs. The transcriptional alterations in nonfunctional
PAs were enriched in autophagy genes, and induction of
autophagy
might
be
beneficial
in
limiting
PA
progression. Indeed, autophagic cell death mediates the
effects of bromocriptine and temozolomide on PA [25–
27]. Nevertheless, the specific roles of autophagy in
antagonizing PA progression remain elusive. Genomic copy number amplifications are associated
with worse prognosis Elevated Ki-67 IHC staining in the PRL-PA high-CNV
group suggests a worse clinical prognosis (Figure 4A
and Table 1). Dopamine agonists (bromocriptine, BCT)
are the first choice for PRL-PA treatment (not applicable
to patients with rapid visual loss and visual field
defects), followed by surgery, which can reduce tumor
size and prolactin levels [4, 24]. However, drug
resistance arises in a subset of patients, as indicated by a
maintained prolactin level or tumor size. The frequency
of BCT resistance in patients in the high-CNV group
(BCT resistance: 10/12 v.s. BCT sensitive: 1/6; p =
0.006, chi-square test) was significantly higher than that
in the low-CNV group (Figure 4D). The Ki-67 label
index marks aggressive behavior in pituitary tumors, and
the high-CNV group (Ki-67 positive ≥3%: 10/16 vs. Ki-
67 positive < 3%: 1/8; P value = 0.02, chi-square test)
was associated with a higher Ki-67 label index (Figure
4E). These data collectively support that copy number
variations contribute to a worse prognosis in PRL-PA
patients. Consistent with previous reports, we observed a lack of
high-frequency recurrent mutations in PAs [12]. However, we do demonstrate frequent genomic copy
number amplifications in PRL-PAs. Moreover, these
genomic CNVs in PRL-PAs play a causal role in PA
etiology, including prolactin production, proliferative
ability, and drug resistance. The impact of copy number
amplifications on PA etiology might be caused by its
direct
influence
on
abnormal
gene
expression
upregulation, such as genes in ribosome biogenesis,
growth signaling and HIF signaling pathways, which
facilitate the hypoxic adaptative ability of PAs. The copy
number amplification and upregulated expression of
BCAT1 may play a central role in the activation of genes
downstream of the mTOR pathway through a feedback
loop involving prolactin [28, 29]. In addition, PRL-PA-
specific upregulation of genes caused by copy number
amplification includes chromogranin B (CHGB), which
has been linked to tumor progression in PRL-PAs [30],
further supporting the role of genomic copy number
variation in PA progression. The transcriptional signature related to relapse in PAs
was determined by comparing patients who experienced
early relapse (< 36 months) and those without any sign
of relapse after a long period (> 60 months). Gene
expression patterns in the high-CNV group in PRL-PA
patients correlated with gene expression patterns that
predict early relapse (Figure 4F), suggesting that the
genes regulated by aberrant copy number in PRL-PAs
are related to relapse. DISCUSSION Altogether,
we
investigated
the
genomic
and
transcriptomic correlation in PAs in the present study
and demonstrated for the first time that high CNVs in
PRL-PAs play an important role in tumor development
and are significantly associated with poor prognosis. We believe the findings have clinical relevance in
defining prognostic subgroups as well as implications
for developing new targeted treatments for PRL-PA. In this study, we profiled transcriptomic alterations in
three major subtypes of Pas revealing shared and
subtype-specific alterations. The clinical subtypes
of PAs were well separated according to their
transcriptional
signatures
in
clustering
analysis,
suggesting that gene expression abnormalities are an
essential part of PA etiology. Interestingly, we found
that the PA subtypes with abnormal hormonal secretion
were more similar to normal pituitary tissue samples
and that the number of DEGs between samples of
these functional subtypes and normal pituitary tissues
samples was less than that of functional subtypes. These findings suggest that the nonfunctional subtype
of PA undergoes more widespread gene expression Weighted
gene
expression
network
analysis
(WGCNA) The PA coexpression network was constructed using the
R
package
WGCNA
(v
1.69)
[22]. Biweight
midcorrelations between each gene pair were calculated
to build an adjacency matrix using the formula:
adjacency = (0.5 * (1+cor) )^power. A thresholding
power of 14 was chosen, and the resulting adjacency
matrix was converted to a topological overlap (TO)
matrix via the TOM similarity algorithm. The genes were
hierarchically clustered based on TO similarity. Modules
were assigned by the dynamic tree-cutting algorithm with
default parameters. Modules were correlated with each
group of samples to identify subtype-specific modules,
and the correlation coefficients and P values are indicated
in figures. Pathway enrichment analysis KEGG pathway enrichment for functional analysis of
gene lists was performed using the clusterprofiler
package under R software (version 3.6.0) [36]. The
significance of the enrichment analysis was defined using
a hypergeometric test, and the resulting P values were
corrected for multiple hypothesis testing with the BH
(Benjamini & Hochberg, 1995) method [37]. The final
reported enriched terms and pathways were filtered
according to adjusted P values < 0.05 or P value < 0.05. Patients samples All samples were obtained following transsphenoidal
surgery performed at Beijing Tiantan Hospital from
May 2013 to May 2017. The fresh tumor samples were AGING 1283 www.aging-us.com High-throughput sequencing A total amount of 0.6 μg genomic DNA per sample was
used as input material for DNA sample preparation. Sequencing libraries were generated using the Agilent
SureSelect
Human
All
Exon
V6
kit
(Agilent
Technologies, CA, USA) following the manufacturer’s
recommendations, and index codes were added to each
sample. Clustering of the index-coded samples was
performed using a cBot Cluster Generation System with
a HiSeq PE Cluster Kit (Illumina, San Diego, CA, USA)
according to the manufacturer’s instructions. After
cluster generation, the DNA libraries were sequenced
using the Illumina HiSeq platform, and 150-bp paired-
end reads were generated. A total amount of 2 μg RNA
per sample was used as input material for RNA sample
preparations. Sequencing libraries were generated using
NEBNext® UltraTM RNA Library Prep Kit for
Illumina® (NEB, Ispawich, USA) following the
manufacturer’s recommendations, and index codes were
added to attribute sequences to each sample. Clustering
of the index-coded samples was performed on a cBot
Cluster Generation System using TruSeq PE Cluster Kit
v3-cBot-HS (Illumina) according to the manufacturer’s
instructions. After cluster generation, the library
preparations were sequenced using the Illumina HiSeq
platform, and 125-bp/150-bp paired-end reads were
generated. Transcriptomic
analysis
and
identification
of
differentially expressed genes For RNA sequencing data, the paired-end clean reads
were aligned to the human reference genome (hg19)
using Hisat2 (v2.0.5) [31]. HTSeq (v 0.11.2) was
used to count the read numbers mapped to each gene
[32]. Hierarchical
clustering
analysis
of
all
transcriptomic samples was performed using the
pairwise similarity of each pair of samples determined
by the Spearman correlation coefficient. Analysis of
differentially expressed genes (DEGs) in each pair of
comparisons was performed using the R package
DESeq2 [33]. The P value of the differential test was
corrected by a multiple hypothesis test, and DEGs were
determined by controlling the FDR (false discovery
rate) < 0.05. Genomic analysis stored in liquid nitrogen. 28 PRL-PAs, 11 GH-PAs and
37 NFPAs from the study population (age range, 20–75
years) were diagnosed according to clinical features and
pathological findings. 9 normal pituitary glands were
obtained from a donation program. The study protocols
were approved by the Internal Review Board of Beijing
Tiantan
Hospital
affiliated
to
Capital
Medical
University and conformed to the ethical guidelines of
the Declaration of Helsinki (No. KY-2013-02). For WGS data, valid sequencing data were mapped to
the reference human genome (UCSC hg19) by Burrows-
Wheeler Aligner (BWA) software to obtain the original
mapping results stored in BAM format [34]. If one or
one paired read(s) were mapped to multiple positions,
the strategy adopted by BWA is to choose the most
likely placement. If two or more likely placements are
presented, BWA selects one randomly. SAMtools and
Picard (http://broadinstitute.github.io/picard/) were used
to sort BAM files and perform duplicate marking, local
realignment, and base quality recalibration to generate
the final BAM file. GATK (v3.4) software 28 was
employed for SNP calling. Genome regions with
significant amplification or deletion in the samples were
evaluated by GISTIC [35], and regions with high
frequencies were screened, namely, recurrent CNV
regions. The higher the GISTIC score is, the higher is
the CNV frequency in this area. FUNDING This study was financially supported by the National
Natural Science Foundation of China (81771489). ACKNOWLEDGMENTS The authors thank the laboratory technicians, data
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PMID:27941783 AGING 1287 www.aging-us.com SUPPLEMENTARY MATERIALS Supplementary Figure 1. Comparison of the number of upregulated DEGs (Red) and downregulated DEGs (Blue) in 6
pairwise DEGs analysis. DESeq2, FDR < 0.05. Supplementary Figure 1. Comparison of the number of upregulated DEGs (Red) and downregulated DEGs (Blue) in 6
pairwise DEGs analysis. DESeq2, FDR < 0.05. AGING 1288 www.aging-us.com Supplementary Figure 2. KEGG pathway enrichment analysis of genes among three PA subtypes vs. Normal. Adjusted P value <
0.05. Supplementary Figure 2. KEGG pathway enrichment analysis of genes among three PA subtypes vs. Normal. Adjusted P value <
0 05 GG pathway enrichment analysis of genes among three PA subtypes vs. Normal. Adjusted P value < AGING 1289 www.aging-us.com www.aging-us.com Supplementary Figure 3. KEGG pathway enrichment analysis of genes in the modules significantly associated with different
PA subtypes. Adjusted P value < 0.05. Supplementary Figure 3. KEGG pathway enrichment analysis of genes in the modules significantly associated with different
PA subtypes. Adjusted P value < 0.05. AGING 1290 www.aging-us.com Supplementary Table Supplementary Table 1. Clinical features of 76 PAs. Supplementary Table 1. Clinical features of 76 PAs. www.aging-us.com 1 Tumor classification by volume, micro: <1 cm, large: 1-4 cm, giant: >4 cm.
2 2 Patients were lost to follow-up: - 2 Patients were lost to follow-up: - 1 Tumor classification by volume, micro: <1 cm, large: 1-4 cm, giant: >4 cm.
2 https://doi.org/10.3892/mmr.2014.2152
PMID:24736764 Sample
Gender
Age (Y)
Clinical Type
Volume1
Invasive/non-Invasive
PFS Time2
Relapse2
Sequencing
strategy
P906
F
25
PRL-PAs
Large
Invasive
81
N
WGS+RNA-seq
P918
M
49
PRL-PAs
Large
non-Invasive
80
N
WGS+RNA-seq
P1133
F
61
PRL-PAs
Large
non-Invasive
66
N
WGS+RNA-seq
P1154
F
46
PRL-PAs
Giant
Invasive
65
N
WGS+RNA-seq
P1037
M
43
PRL-PAs
Giant
Invasive
37
Y
WGS+RNA-seq
P1408
M
45
PRL-PAs
Giant
non-Invasive
52
N
WGS
P1100
M
37
PRL-PAs
Large
non-Invasive
41
Y
WGS
P1694
F
44
PRL-PAs
Large
Invasive
39
N
WGS+RNA-seq
P1711
F
30
PRL-PAs
Large
non-Invasive
-
-
WGS+RNA-seq
P1712
M
31
PRL-PAs
Giant
Invasive
8
Y
WGS+RNA-seq
P1736
M
55
PRL-PAs
Giant
Invasive
17
Y
WGS+RNA-seq
P1809
F
20
PRL-PAs
Large
Invasive
34
N
WGS+RNA-seq
P25502
M
65
PRL-PAs
Large
Invasive
-
-
WGS+RNA-seq
P28607
F
28
PRL-PAs
Large
Invasive
-
-
WGS+RNA-seq
P2890
F
15
PRL-PAs
Large
non-Invasive
-
-
WGS+RNA-seq
P1824
M
22
PRL-PAs
Giant
Invasive
33
N
WGS+RNA-seq
P25505
M
27
PRL-PAs
Large
Invasive
-
-
WGS
P1821
F
22
PRL-PAs
Large
non-Invasive
-
-
WGS
P1070
F
50
PRL-PAs
Large
non-Invasive
71
N
WGS+RNA-seq
P961
M
42
PRL-PAs
Giant
Invasive
53
Y
WGS+RNA-seq
P1483
M
20
PRL-PAs
Giant
Invasive
49
N
WGS+RNA-seq
P1587
F
26
PRL-PAs
Large
Invasive
44
N
WGS+RNA-seq
P_N6_30
F
48
PRL-PAs
Large
non-Invasive
52
N
WGS
P1199
F
41
PRL-PAs
Large
Invasive
4
Y
WGS+RNA-seq
P1144
M
48
PRL-PAs
Large
Invasive
66
N
WGS
P1825
F
38
PRL-PAs
Giant
Invasive
33
N
WGS+RNA-seq
P640829
M
43
PRL-PAs
Large
Invasive
-
-
WGS+RNA-seq
P29115
M
37
PRL-PAs
Large
Invasive
-
-
WGS
P1087
M
46
NFPAs
Giant
Invasive
25
Y
WGS
P1169
F
57
NFPAs
Giant
Invasive
65
N
WGS
P1271
M
56
NFPAs
Large
non-Invasive
59
N
WGS+RNA-seq
P1301
F
55
NFPAs
Large
non-Invasive
14
Y
WGS
P1315
F
50
NFPAs
Large
non-Invasive
57
N
WGS
P1339
M
43
NFPAs
Large
Invasive
55
N
WGS+RNA-seq
P1356
M
47
NFPAs
Large
Invasive
36
Y
WGS+RNA-seq
P1391
M
51
NFPAs
Large
non-Invasive
52
N
WGS
P1409
M
60
NFPAs
Large
Invasive
52
N
WGS
P1423
F
66
NFPAs
Large
non-Invasive
51
N
WGS+RNA-seq
P1448
F
49
NFPAs
Giant
Invasive
44
Y
WGS+RNA-seq
P1454
F
71
NFPAs
Giant
non-Invasive
50
N
WGS+RNA-seq
P1467
F
54
NFPAs
Giant
Invasive
24
Y
WGS+RNA-seq
P1487
M
65
NFPAs
Giant
Invasive
49
N
WGS+RNA-seq
P1574
M
65
NFPAs
Giant
Invasive
3
Y
WGS+RNA-seq
P1582
M
66
NFPAs
Giant
Invasive
45
N
WGS+RNA-seq
P1594
M
67
NFPAs
Large
non-Invasive
44
N
WGS
P1605
F
44
NFPAs
Large
non-Invasive
44
N
WGS+RNA-seq
P1613
M
51
NFPAs
Large
Invasive
43
N
WGS
P1643
M
43
NFPAs
Large
non-Invasive
36
Y
WGS+RNA-seq
P_N6_15
M
49
NFPAs
Giant
Invasive
55
N
WGS+RNA-seq
P_N6_16
M
34
NFPAs
Giant
Invasive
55
N
WGS+RNA-seq
P_N6_20
M
61
NFPAs
Giant
Invasive
54
N
WGS+RNA-seq
P_N6_21
F
51
NFPAs
Large
Invasive
54
N
WGS+RNA-seq
P_N6_29
M
47
NFPAs
Large
non-Invasive
52
N
WGS+RNA-seq AGING 1291 www.aging-us.com P_N6_32
M
64
NFPAs
Giant
Invasive
52
N
WGS+RNA-seq
P_N6_34
M
55
NFPAs
Giant
Invasive
52
N
WGS+RNA-seq
P_N6_4
F
32
NFPAs
Large
Invasive
57
N
WGS+RNA-seq
P_N6_48
M
64
NFPAs
Giant
Invasive
51
N
WGS+RNA-seq
P_N6_5
F
35
NFPAs
Giant
Invasive
57
N
WGS+RNA-seq
P_N6_50
F
41
NFPAs
Large
Invasive
51
N
WGS
P_N6_56
M
45
NFPAs
Large
Invasive
51
N
WGS
P_N6_61
F
49
NFPAs
Giant
Invasive
49
N
WGS+RNA-seq
P1183
F
57
NFPAs
Giant
Invasive
66
N
WGS
P1068
M
48
NFPAs
Giant
Invasive
19
Y
WGS
P1182
M
67
NFPAs
Large
Invasive
66
N
WGS
P1195
M
35
NFPAs
Large
non-Invasive
65
N
WGS
P1284
F
21
GH-PAs
Giant
Invasive
12
Y
WGS+RNA-seq
P1298
F
69
GH-PAs
Large
non-Invasive
57
N
WGS+RNA-seq
P1332
M
57
GH-PAs
Giant
Invasive
14
N
WGS+RNA-seq
P1352
M
31
GH-PAs
Giant
Invasive
3
Y
WGS+RNA-seq
P23
F
51
GH-PAs
Giant
Invasive
4
Y
WGS+RNA-seq
P46
F
44
GH-PAs
Large
non-Invasive
51
N
WGS+RNA-seq
P1603
M
45
GH-PAs
Large
non-Invasive
44
N
WGS+RNA-seq
P1563
M
32
GH-PAs
Large
Invasive
12
Y
WGS
P1547
F
33
GH-PAs
Large
non-Invasive
46
N
WGS+RNA-seq
P1520
M
21
GH-PAs
Giant
Invasive
17
Y
WGS+RNA-seq
P1660
F
51
GH-PAs
Large
non-Invasive
41
N
WGS+RNA-seq
1 Tumor classification by volume micro: <1 cm large: 1 4 cm giant: >4 cm AGING 1292 www.aging-us.com Supplementary Material Supplementary Material Supplementary Material 1. Sense mutations of CDS in 11 pairs GH-PAs. Supplementary Material 1. Sense mutations of CDS in 11 pairs GH-PAs. Please browse Full Text version to see the data of Supplementary Material 1. AGING 1293 www.aging-us.com
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https://openalex.org/W2157406619
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Open Science
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Inhibition of β-catenin/p300 interaction proximalizes mouse embryonic lung epithelium
|
Taro Sasaki
|
English
|
Spoken
| 8,990
| 16,652
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* Correspondence: kahnm@usc.edu
1Department of Biochemistry and Molecular Biology, Keck School of
Medicine, University of Southern California, 1450 Biggy Street, Los Angeles,
CA 90033, USA
2Center for Molecular Pathways and Drug Discovery, Keck School of
Medicine, University of Southern California, 1450 Biggy Street, Los Angeles,
CA 90033, USA
Full list of author information is available at the end of the article © 2014 Sasaki and Kahn; licensee Springer. This is an Open Access article distributed under the terms of the Creative Commons
Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction
in any medium, provided the original work is properly credited. Abstract Background: Wnt/β-catenin signaling has been suggested to regulate proximal-distal determination of embryonic
lung epithelium based upon genetically modified mouse models. The previously identified and characterized small
molecule inhibitor IQ1 can pharmacologically decrease the interaction between β-catenin and its transcriptional
coactivator p300, thereby enhancing the β-catenin/CBP interaction. Inhibition of the β-catenin/p300 interaction by
IQ1 blocks the differentiation of embryonic stem cells and epicardial progenitor cells; however, whether differential
coactivator usage by β-catenin plays a role in proximal-distal determination of lung epithelium is unknown. Methods: We examined the effects of inhibiting the β-catenin/p300 interaction with IQ1 on lung branching
morphogenesis in mouse embryos in utero and mouse embryonic lung organ culture ex vivo. The phenotype of
IQ1 treated lungs was analyzed by epithelial staining, histology, quantitative PCR and in situ hybridization. Results: Inhibition of the β-catenin/p300 interaction by IQ1 disrupted the distal branching of mouse lung
epithelium both in utero and ex vivo. IQ1 proximalized lung epithelium with decreased expression of the genes
Bmp4 and Fgf10, hallmarks of distal lung determination, and increased expression of the proximal genes Sox2 and
Scgb1a1 (CC10) as shown by quantitative PCR and in situ hybridization. The disruption of branching was reversible
ex vivo as branching was reinitiated after removal of IQ1 from the media. Conclusions: The results demonstrate that the β-catenin/p300 interaction plays a critical role in proximal-distal
determination of the epithelium in mouse lung branching morphogenesis and β-catenin/p300 inhibition
pharmacologically proximalizes lung epithelium. Keywords: Lung; Branching; Differentiation; Wnt; β-catenin; p300; CBP; IQ1; ICG-001; Small molecule inhibitor fibrotic complications [3]. Regeneration is in essence a
recapitulation of development. Therefore, understanding
the signaling pathways and decision points that control
lung development is a critical goal. Lung development in-
volves branching morphogenesis. The primordial epithelial
tube undergoes repetitive branching at distal tips resulting
in a tree-like structure. Lung branching morphogenesis is
regulated by multiple signaling pathways including the FGF
pathway [4] and the Wnt signaling pathway [5]. β-catenin
is the central signaling mediator of canonical Wnt sig-
naling. Therefore, β-catenin has repeatedly been genet-
ically deleted for loss-of-function analysis or stabilized
for gain-of-function analysis in mice to study Wnt
signaling. During lung development, tissue-specific
deletion of β-catenin in the epithelium was found to
proximalize the epithelium by expanding the proximal
airway and inhibiting distal airway growth [6]. Based Inhibition of β-catenin/p300 interaction
proximalizes mouse embryonic lung epithelium Tomoyo Sasaki1 and Michael Kahn1,2,3* Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Open Access © 2014 Sasaki and Kahn; licensee Springer. This is an Open Access article distributed under the terms of the Creative Common
Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction
in any medium, provided the original work is properly credited. Compounds IQ1 and ICG-001 were synthesized in our laboratory
as previously described [7,11]. IQ1 and ICG-001 were
dissolved in dimethyl sulfoxide (DMSO) to prepare 100 mM
or 500 mM stock solutions. Equal volumes of DMSO
were used for controls. Chemical genetic approaches offer a complementary
method to genetic manipulation, to selectively modulate
the transcriptional role of β-catenin. To activate transcrip-
tion, β-catenin must translocate to the nucleus to interact
with transcription factors e.g. members of the TCF/LEF in
canonical Wnt signaling as well as various transcriptional
co-activators. We have identified and characterized a
number of small molecules that specifically inhibit the
protein-protein interaction between β-catenin and either of
its two Kat3 coactivators i.e. Creb binding protein (CBP) or
p300. Specifically inhibiting either the β-catenin/CBP
or the β-catenin/p300 interaction does not disturb
other protein-protein interactions involving β-catenin,
including the β-catenin/E-Cadherin interaction that is
critically responsible for epithelial cell-cell adhesion. Therefore, specific well-characterized small molecule
pharmacologic tools provide an advantage for loss-of-
function analysis of multifunctional proteins like β-catenin. An additional potential advantage of utilizing a pharmaco-
logic approach is the ability to temporally utilize small
molecule pharmacologic agents to manipulate cell prolif-
eration and differentiation for the purpose of regenerative
medicine. IQ1 (C21H22N4O2, molecular weight 362.42)
and ICG-001 (C33H32N4O4, molecular weight 548.63) are
specific inhibitors of the interaction between β-catenin and
the transcriptional coactivators p300 and CBP respectively. IQ1 inhibits the interaction between β-catenin and p300
indirectly via targeting the PR72/130 subunit of PP2A
and thereby blocking PP2A/Nkd complex formation [7]. On the other hand, ICG-001 directly inhibits the inter-
action between β-catenin and CBP via binding to the
amino terminus of CBP [8]. Using these small mole-
cules, we have previously demonstrated the distinct
roles that the two coactivators p300 and CBP in β-catenin
dependent transcriptional gene regulation, despite their
high degree of identity and even higher homology. Inhibition of the β-catenin/p300 interaction by IQ1
prevented the differentiation of embryonic stem cells
[7], whereas inhibition of β-catenin/CBP interaction
enhanced the differentiation of neural progenitor cells [9]
and human embryonic stem cells (hES cells) [10]. However,
the role of β-catenin differential coactivator utilization
in lung development and branching morphogenesis is
unknown Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Page 2 of 10 To investigate the role of β-catenin differential coactivator
utilization in murine lung development and branching mor-
phogenesis, we treated mouse embryonic lungs with IQ1 in
utero and ex vivo to selectively inhibit the β-catenin/p300
interaction and analyzed the phenotype by epithelial stain-
ing, histology, quantitative PCR and in situ hybridization. on this analysis, β-catenin was suggested to regulate
proximal-distal determination of the epithelium during
lung morphogenesis. However, beyond its role as a
transcriptional activator of Wnt signaling, β-catenin is also
an important protein component of adherens junction,
which is critically responsible for epithelial cell-cell adhe-
sion. Thus, genetic deletion of β-catenin in mice results in
both disruption in cell-cell adhesion and the transcrip-
tional role of β-catenin, making analysis of the phenotype
complex [5]. on this analysis, β-catenin was suggested to regulate
proximal-distal determination of the epithelium during
lung morphogenesis. However, beyond its role as a
transcriptional activator of Wnt signaling, β-catenin is also
an important protein component of adherens junction,
which is critically responsible for epithelial cell-cell adhe-
sion. Thus, genetic deletion of β-catenin in mice results in
both disruption in cell-cell adhesion and the transcrip-
tional role of β-catenin, making analysis of the phenotype
complex [5]. Mice TOPGAL mice were obtained from The Jackson Laboratory. Vaginal plug at noon was considered as day 0.5 of preg-
nancy (E0.5). For administration of IQ1 or ICG-001 to
embryos in utero, timed pregnant mice were fed IQ1
(72 or 144 mg/kg/day) or ICG-001 (220 mg/kg/day)
mixed in peanut butter in addition to their regular diet. The animal study protocol was approved by the IACUC at
the University of Southern California. Whole mount NBT/BCIP and β-galactosidase staining
Lung samples were fixed in 4% paraformaldehyde for
1 hour to overnight and washed in phosphate buffered
saline (PBS). The lungs were incubated in a solution of
NBT/BCIP Ready-to-Use Tablets (Roche Applied Science)
according to the manufacturer’s directions. After develop-
ment of color, excess staining was removed by dehydration
and rehydration through a series of methanol washes. The stained lungs were cleared in graded steps with
glycerol before photographing. Branching tips of the
stained lungs were counted under a bright-field stereo-
microscope. For histological analysis, the stained lungs
were re-fixed in 4% paraformaldehyde, embedded in
paraffin, sectioned and counterstained with Nuclear Fast
Red (Electron Microscopy Sciences). For β-galactosidase
staining, lungs were dissected, fixed in 0.5% glutaralde-
hyde with 2 mM of magnesium chloride and stained as
described previously. The images were captured using an
AxioImager Z1 and AxioVision digital image processing
software (Carl Zeiss) or SZX7 stereomicroscope and infin-
ity2 imaging system (Olympus). Background The ability to regulate proliferation and differentiation
of the lung has important consequences in regards to
both premature birth and bronchopulmonary dysplasia [1]
and diseases of the adult lung i.e. idiopathic pulmonary
fibrosis [2]. Furthermore, within the context of regen-
erative medicine, the ability to control lung develop-
ment may replace conventional lung transplantation,
which has inevitable problems associated with both a
shortage of suitable donors and immunogenicity leading
to the requirement for long term immunosuppression and Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Whole mount in situ hybridization The lung explants were fixed in 4% paraformaldehyde for
4 hours to overnight, stored in 100% methanol at −20°C,
bleached in 3% hydrogen peroxide in methanol, rehydrated
and subsequently underwent in situ hybridization as previ-
ously described. Digoxigenin-labeled anti-sense RNA probes
were synthesized from subcloned mouse gene templates
using DIG RNA Labeling Kit (Roche Applied Science). Hybridization was colorized with ALP conjugated anti-
DIG antibody (Roche Applied Science) and NBT/BCIP
Ready-to-Use Tablets (Roche Applied Science). Experiments
were independently repeated at least three times. The
plasmids including cDNA of mouse Nkx2-1 and Bmp4
were generously provided by Dr. Changgong Li. Lung organ culture Lungs were dissected from mouse embryos in cold sterile
Hanks’ Balanced Salt Solution under a stereomicroscope. The lungs were randomized by size and obvious outliers
were excluded. The lungs were placed on a polycarbonate
membrane filter (Whatman) floating on 1 ml of serum-free
BGJb medium (Invitrogen) with penicillin/streptomycin in Page 3 of 10 Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Page 3 of 10 Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Laboratories) and anti-α-SMA (1:200, Sigma). For whole
mount PECAM-1 staining, fixed lungs were dehydrated
in methanol, bleached in 3% hydrogen peroxide in in
methanol, rehydrated, blocked in 1% BSA, incubated
with anti-PECAM-1 antibody (1:200, Santa Cruz, sc-1506)
overnight at 4°C, washed in PBS, incubated with HRP-
conjugated anti-goat IgG (Santa Cruz) and colorized
with diaminobenzidine (DAB) substrate (Sigma). Laboratories) and anti-α-SMA (1:200, Sigma). For whole
mount PECAM-1 staining, fixed lungs were dehydrated
in methanol, bleached in 3% hydrogen peroxide in in
methanol, rehydrated, blocked in 1% BSA, incubated
with anti-PECAM-1 antibody (1:200, Santa Cruz, sc-1506)
overnight at 4°C, washed in PBS, incubated with HRP-
conjugated anti-goat IgG (Santa Cruz) and colorized
with diaminobenzidine (DAB) substrate (Sigma). 60 mm Center-well Organ Culture Dishes (BD Falcon). The lung explants were positioned at the gas–liquid
interface on the membrane filters. The Cultures were
kept at 37°C in 5% CO2 and media was changed daily. The β-catenin/p300 interaction during lung branching
morphogenesis To investigate the role of differential β-catenin coactivator
usage during lung branching morphogenesis, we used
the coactivator selective small molecule inhibitors of the
β-catenin/p300 [7] and β-catenin/CBP [8] interaction
that we previously identified and have utilized extensively
[2,7,9,12]. To inhibit the β-catenin/p300 interaction during
lung morphogenesis, we treated E12.5 mouse embryos with
IQ1 in utero for two days via oral administration of IQ1 to
their pregnant mothers. In the embryonic mice at day
E14.5, we found that the inhibition of the β-catenin/p300
interaction disrupted lung branching morphogenesis. IQ1
decreased the size of the lung and dramatically decreased
the number of branching tips (Figure 2A). Despite the
branching inhibition at the distal tips, IQ1 caused an
elongation of the proximal airway compared to the control
lung at E14.5 (Figure 2A). This elongation of the proximal Alkaline phosphatase activity marks murine lung
epithelium during branching morphogenesis Alkaline phosphatase activity marks murine lung
epithelium during branching morphogenesis p
g
g
p
g
Wnt signaling apparently plays a critical role during lung
branching morphogenesis. We found that murine em-
bryonic lung expresses endogenous alkaline phosphat-
ase (ALP) activity in the epithelium. The epithelium can
be selectively stained utilizing the endogenous ALP activ-
ity with the chromogenic substrate, nitro-blue tetrazolium
chloride
and
5-bromo-4-chloro-3′-indolyphosphate
p-toluidine salt (NBT/BCIP). We therefore sought to
utilize NBT/BCIP to visualize epithelial branching mor-
phogenesis in whole mount mouse lung. In the event,
we stained mouse lung at four different embryonic stages
from E11.5 to E14.5 with NBT/BCIP. We observed
blue-purple staining specifically visualizing lung epi-
thelial structures during branching morphogenesis at
all four time points (Figure 1A). Next, we sectioned the
stained lungs for histological analysis and confirmed
that the blue-purple staining was localized on the apical
surface of the lung epithelium and not in the mesenchyme
(Figure 1B and 1C). These results suggest that endogenous
ALP activity is a specific marker for embryonic mouse lung
epithelium and NBT/BCIP, chromogenic substrate of ALP,
can be used to visualize the epithelial branching structure
in whole mount mouse lung using bright-field microscopy. Quantitative PCR (qPCR) The lung explants were homogenized with TRIzol® reagent
(Invitrogen) by passing through a 25-gage needle and total
RNA was isolated according to the manufacturer’s instruc-
tions. The concentration of total RNA was measured using
a NanoDrop® spectrophotometer and reverse transcrip-
tion was performed with an iScript cDNA Synthesis Kit
(Bio-Rad). Quantitative PCR amplification was performed
using SYBR Green PCR master mix reagent (BioRad) and
a MyiQ thermal cycler (BioRad) with the following gene-
specific primers: mouse Bmp4 (CGTTACCTCAAGGGAG
TCGAGATTG, TCTTATTCTTCTTCCTGGACCGCTG),
mouse Fgf10 (TGCACATACATGAGCCCTTTGT, TTTG
CTCAGGTTAAGCCCCAG), mouse Nkx2-1 (AAATTTG
GGGGTCTTTCTGG, AGAGTGCATCCACAGGGAAG),
mouse Shh (ACTCACCCCCAATTACAACCC, TGCTCC
CGTGTTTTCCTCA), mouse Scgb1a1 (CAGCTGAAGA
GACTGGTGGAT, TGTTAGATTTTCTCCGTGAGCTT). Melt curve analysis and gel electrophoresis were used to
examine the specificity of amplified products. Data were
normalized to the reference gene, mouse Gapdh. Relative
expression levels were calculated using the 2^-ddCt
method. Data are presented as mean ± SD. Differences in
means between two experimental groups were analyzed
using two-sample, two-tailed Student’s t-test. p < 0.05 was
considered significant. Histology and immunohistochemistry Lung samples were fixed in 4% paraformaldehyde overnight,
embedded in OCTcompound or paraffin and sectioned. For
immunohistochemistry, the sections were blocked in 1%
bovine serum albumin (BSA) (Jackson ImmunoResearch)
and incubated with primary antibodies. The antibodies used
were FITC mouse anti-β-catenin (1:400, BD Transduction Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
Page 4 of 10
http://www.transrespmed.com/content/2/1/8 Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Page 4 of 10 Figure 1 Alkaline phosphatase (ALP) activity marks epithelial branching structures in embryonic mouse lung. (A) Mouse embryonic
lungs from four different embryonic stages i.e. E11.5, E12.5, E13.5 and E14.5 were stained with ALP substrate NBT/BCIP. Endogenous ALP activity
in blue-purple visualizes epithelial branching morphogenesis in the whole-mount lungs. (B) Histology of the stained E13.5 lung shows the specific
ALP activity around the airway cavities (*), but not in the mesenchyme. (C) Higher magnification shows that the ALP activity is specific to the
apical surface of the lung epithelial cells (arrows). Figure 1 Alkaline phosphatase (ALP) activity marks epithelial branching structures in embryonic mouse lung. (A) Mouse embryonic
lungs from four different embryonic stages i.e. E11.5, E12.5, E13.5 and E14.5 were stained with ALP substrate NBT/BCIP. Endogenous ALP activity
in blue-purple visualizes epithelial branching morphogenesis in the whole-mount lungs. (B) Histology of the stained E13.5 lung shows the specific
ALP activity around the airway cavities (*), but not in the mesenchyme. (C) Higher magnification shows that the ALP activity is specific to the
apical surface of the lung epithelial cells (arrows). Figure 2 Inhibition of the β-catenin/p300 interaction disrupts branching morphogenesis in mouse lung. (A) Lungs from embryonic mice
treated with the β-catenin/p300 inhibiter IQ1 (right) or DMSO control (left) in utero. Pregnant mice received IQ1 (72 mg/kg/day) orally to
expose the embryos to IQ1 in utero from E12.5 for 2 days (E12.5 + 2). The lungs of the embryos were dissected and the epithelium was stained with
the ALP substrate NBT/BCIP. IQ1 decreased the size of the lung but elongated the airways. (B) Pregnant mice received IQ1 (72 mg/kg/day for low dose,
144 mg/kg/day for high dose) or ICG-001 (220 mg/kg/day) orally to expose the embryos in utero from E9.5 for 3 days (E9.5 + 3). IQ1 inhibited lung branching
morphogenesis but ICG-001 did not. (C) H-E staining shows that IQ1 dilated the airways and decreased the density of mesenchymal cells. Inhibition of β-catenin/p300 interaction proximalizes the
lung epithelium The elongated proximal airways observed in the IQ1
treated lungs led us to investigate the hypothesis that
specific inhibition of the β-catenin/p300 interaction
proximalizes the lung epithelium. To test this hypothesis,
we analyzed the expression of both distal and proximal
genes in lung explants by quantitative PCR (qPCR). We
selected Bmp4, Fgf10 and Nkx2-1 (Ttf-1) as distal marker
genes and Sox2 and Scgb1a1 (CC10) as proximal marker
genes. The critical growth factors involved in branching
morphogenesis (Bmp4, Fgf10, Shh) were examined at 6 hours
and 24 h. As anticipated, IQ1 significantly decreased the
expression of the distal genes, Bmp4, Fgf10 and Nkx2-1
(Figure 4A). In addition, whole mount in situ hybridization
confirmed the decrease of the distal genes Bmp4 and
Nkx2-1 expressed in the distal epithelium (Figure 4B). At same time, IQ1 significantly increased the expression
of the proximal genes, Sox2 and Scgb1a1 (Figure 4A). However, IQ1 did not decrease the expression of the
widely expressed lung epithelial marker Shh, suggesting
that IQ1 did not disrupt the overall growth of the lung
epithelium (Figure 4A). Our results indicate that IQ1 decreased the expression of distal genes and increased the
expression of proximal genes in the lung explants consist-
ent with the hypothesis that inhibition of the β-catenin/
p300 interaction proximalizes lung epithelium. Histology and immunohistochemistry Next, we applied IQ1 to lung organ culture to confirm
that the disruption of branching in utero did not result
from overall poor general condition of the embryos, as
we have previously demonstrated that IQ1 also affects
cardiac and vascular system development [13]. We cultured
E12.5 lung explants at a gas–liquid interface on membrane
filters for 24 hours. We examined the effects of both the
specific small molecule β-catenin/CBP inhibitor ICG-001,
as well as the β-catenin/p300 inhibitor IQ1 in this model. We found that inhibition of the β-catenin/p300 interaction
with IQ1, but not the β-catenin/CBP interaction with
ICG-001, significantly decreased the number of distal
tips at 24 hours ex vivo (Figure 2D and 2E). The differ-
ences are even more striking after two days of culture
(Figure 3A). ICG-001 slightly slowed the growth of
explants, however we observed new formation of distal
tips in ICG-001 treated but not in IQ1 treated cultures
(Figure 2D and 3A). The effects of differential coactivator
modulation in lung organ culture confirmed that the dis-
ruption of branching morphogenesis in utero was a direct
effect on the lung and did not result from overall inhib-
ition of embryonic development. Our results suggest that
the β-catenin/p300 interaction plays a critical role during
mouse lung branching morphogenesis. Figure 3 Histological analyses of IQ1 treated lungs. (A) TOPGAL,
the Wnt signaling reporter mouse shows activation of β-catenin/LEF/
TCF reporter by expression of β-galactosidase. E12.5 lung explants of
TOPGAL mouse were treated with either DMSO (control), IQ1 (10 μM)
or ICG-001 (10 μM) for 48 hours. β-galactosidase (blue) activity is
observed in the bronchial epithelium of the proximal airway (arrows). IQ1 did not decrease β-galactosidase activity, whereas ICG-001, a
known CBP/catenin antagonist, decreased β-galactosidase activity in
the proximal airway. (B) Immunohistochemistry shows localization of
β-catenin at the cell membrane of epithelial cell (arrows) and
mesenchymal cells. (C) Whole mount PECAM-1 staining (brown)
shows capillaries around the distal tips. Four days of IQ1 treatment in
utero did not significantly disrupt formation of capillaries as judged by
PECAM staining. Histology and immunohistochemistry (D) E12.5 mouse
embryonic lungs were cultured with either DMSO, IQ1 (10 μM) or ICG-001 (10 μM) for 24 hours and stained with ALP substrate NBT/BCIP. The arrows
indicate new distal tips in DMSO and ICG-001 treated explants. IQ1 disrupted branching morphogenesis ex vivo. (E) IQ1 significantly decreased the number
of the distal tips after 24 hour culture of E12.5 lungs. Data are mean ± SD from five lungs per group. *p < 0.025 vs. control by t-test. Figure 2 Inhibition of the β-catenin/p300 interaction disrupts branching morphogenesis in mouse lung. (A) Lun Figure 2 Inhibition of the β-catenin/p300 interaction disrupts branching morphogenesis in mouse lung. (A) Lungs from embryonic mice
treated with the β-catenin/p300 inhibiter IQ1 (right) or DMSO control (left) in utero. Pregnant mice received IQ1 (72 mg/kg/day) orally to
expose the embryos to IQ1 in utero from E12.5 for 2 days (E12.5 + 2). The lungs of the embryos were dissected and the epithelium was stained with
the ALP substrate NBT/BCIP. IQ1 decreased the size of the lung but elongated the airways. (B) Pregnant mice received IQ1 (72 mg/kg/day for low dose,
144 mg/kg/day for high dose) or ICG-001 (220 mg/kg/day) orally to expose the embryos in utero from E9.5 for 3 days (E9.5 + 3). IQ1 inhibited lung branching
morphogenesis but ICG-001 did not. (C) H-E staining shows that IQ1 dilated the airways and decreased the density of mesenchymal cells. (D) E12.5 mouse
embryonic lungs were cultured with either DMSO, IQ1 (10 μM) or ICG-001 (10 μM) for 24 hours and stained with ALP substrate NBT/BCIP. The arrows
indicate new distal tips in DMSO and ICG-001 treated explants. IQ1 disrupted branching morphogenesis ex vivo. (E) IQ1 significantly decreased the number
of the distal tips after 24 hour culture of E12.5 lungs. Data are mean ± SD from five lungs per group. *p < 0.025 vs. control by t-test. Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
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http://www.transrespmed.com/content/2/1/8 Page 5 of 10 Page 5 of 10 airways suggests that the impact of IQ1 is not due simply
to growth inhibition. IQ1 also inhibited branching with
3 day treatment initiating at E9.9; however, ICG-001
did not affect branching in utero (Figure 2B). Further,
we observed that IQ1 caused airway dilation and de-
creased the density of mesenchymal cells (Figure 2C). Histology and immunohistochemistry Next, we applied IQ1 to lung organ culture to confirm
that the disruption of branching in utero did not result
from overall poor general condition of the embryos, as
we have previously demonstrated that IQ1 also affects
cardiac and vascular system development [13]. We cultured
E12.5 lung explants at a gas–liquid interface on membrane
filters for 24 hours. We examined the effects of both the
specific small molecule β-catenin/CBP inhibitor ICG-001,
as well as the β-catenin/p300 inhibitor IQ1 in this model. We found that inhibition of the β-catenin/p300 interaction
with IQ1, but not the β-catenin/CBP interaction with
ICG-001, significantly decreased the number of distal
tips at 24 hours ex vivo (Figure 2D and 2E). The differ-
ences are even more striking after two days of culture
(Figure 3A). ICG-001 slightly slowed the growth of
explants, however we observed new formation of distal
tips in ICG-001 treated but not in IQ1 treated cultures
(Figure 2D and 3A). The effects of differential coactivator
modulation in lung organ culture confirmed that the dis-
ruption of branching morphogenesis in utero was a direct
effect on the lung and did not result from overall inhib-
ition of embryonic development. Our results suggest that
the β-catenin/p300 interaction plays a critical role during
mouse lung branching morphogenesis. Figure 3 Histological analyses of IQ1 treated lungs. (A) TOPGAL,
the Wnt signaling reporter mouse shows activation of β-catenin/LEF/
TCF reporter by expression of β-galactosidase. E12.5 lung explants of
TOPGAL mouse were treated with either DMSO (control), IQ1 (10 μM)
or ICG-001 (10 μM) for 48 hours. β-galactosidase (blue) activity is
observed in the bronchial epithelium of the proximal airway (arrows). IQ1 did not decrease β-galactosidase activity, whereas ICG-001, a
known CBP/catenin antagonist, decreased β-galactosidase activity in
the proximal airway. (B) Immunohistochemistry shows localization of
β-catenin at the cell membrane of epithelial cell (arrows) and
mesenchymal cells. (C) Whole mount PECAM-1 staining (brown)
shows capillaries around the distal tips. Four days of IQ1 treatment in
utero did not significantly disrupt formation of capillaries as judged by
PECAM staining. airways suggests that the impact of IQ1 is not due simply
to growth inhibition. IQ1 also inhibited branching with
3 day treatment initiating at E9.9; however, ICG-001
did not affect branching in utero (Figure 2B). Further,
we observed that IQ1 caused airway dilation and de-
creased the density of mesenchymal cells (Figure 2C). Proximalizations of the embryonic lung selective via
inhibition of the β-catenin/p300 interaction or β-catenin
deletion are mechanistically distinct (B) Whole mount in situ
hybridization shows local expression of Bmp4 and Nkx2-1 in the distal epithelium. IQ1 decreased the expression of Bmp4 and Nkx2-1 at 24 hours. binding motifs are a hallmark of canonical Wnt target
genes as well as commonly used Wnt signaling reporters. TOPGAL transgenic mice express β-galactosidase under
the control of canonical Wnt TCF/β-catenin transcription. To evaluate the effects of specific inhibition of the
β-catenin/p300 interaction on Wnt signaling in mouse
embryonic lung, we treated lung explants from TOPGAL
reporter mice with IQ1. In the vehicle control treated mice,
we observed activated Wnt/TCF/β-catenin dependent
transcription as judged by β-galactosidase activity at the
bronchial epithelium of the proximal airway (Figure 3A,
DMSO, arrow). Selective inhibition of the β-catenin/p300
interaction with IQ1 did not decrease the β-galactosidase
activity in the proximal airways (Figure 3A, IQ1, arrow),
whereas ICG-001, a known inhibitor of β-catenin/CBP
interaction, decreased the proximal β-galactosidase
activity (Figure 3A, ICG-001, arrow). These results demon-
strate that whereas genetic deletion of β-catenin decreases
overall Wnt/TCF/β-catenin dependent signaling, on the
contrary, selective inhibition of the β-catenin/p300
interaction by IQ1 maintains rather than decreases
Wnt/β-catenin/TCF dependent reporter activity in the
proximal airways. Furthermore, selective inhibition of
the β-catenin/CBP interaction decreases proximal but binding motifs are a hallmark of canonical Wnt target
genes as well as commonly used Wnt signaling reporters. TOPGAL transgenic mice express β-galactosidase under
the control of canonical Wnt TCF/β-catenin transcription. To evaluate the effects of specific inhibition of the
β-catenin/p300 interaction on Wnt signaling in mouse
embryonic lung, we treated lung explants from TOPGAL
reporter mice with IQ1. In the vehicle control treated mice,
we observed activated Wnt/TCF/β-catenin dependent
transcription as judged by β-galactosidase activity at the
bronchial epithelium of the proximal airway (Figure 3A,
DMSO, arrow). Selective inhibition of the β-catenin/p300
interaction with IQ1 did not decrease the β-galactosidase
activity in the proximal airways (Figure 3A, IQ1, arrow),
whereas ICG-001, a known inhibitor of β-catenin/CBP
interaction, decreased the proximal β-galactosidase
activity (Figure 3A, ICG-001, arrow). These results demon-
strate that whereas genetic deletion of β-catenin decreases
overall Wnt/TCF/β-catenin dependent signaling, on the
contrary, selective inhibition of the β-catenin/p300
interaction by IQ1 maintains rather than decreases
Wnt/β-catenin/TCF dependent reporter activity in the
proximal airways. Furthermore, selective inhibition of
the β-catenin/CBP interaction decreases proximal but not distal Wnt/β-catenin/TCF dependent activity without
apparently affecting normal lung development ex vivo. Proximalizations of the embryonic lung selective via
inhibition of the β-catenin/p300 interaction or β-catenin
deletion are mechanistically distinct The proximalization resulting from IQ1 inhibition of the
β-catenin/p300 interaction phenotypically resembles those
seen utilizing genetic deletion of β-catenin [6,14]. There-
fore, we decided to compare in detail the mechanisms
involved in the pharmacologic inhibition of the β-catenin/
p300 interaction to the genetic deletion of β-catenin. β-catenin interacts with many additional transcription
partners beyond p300 and CBP. Amongst these, the
TCF/LEF family of transcription factors has been most
extensively investigated, as consensus LEF1/TCF-DNA Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
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http://www.transrespmed.com/content/2/1/8 Page 6 of 10 Page 6 of 10 Figure 4 Gene expression analyses of IQ1 treated lung explants. (A) Quantitative PCR (qPCR) analysis of the lung explants. Lung explants
were cultured with either DMSO (control), IQ1 (10 μM) or ICG-001 (10 μM) for 6 hours or 24 hours. Relative expression changes compared to
E12.5 lungs without culture (0 h) are presented. IQ1 decreased the expression of Bmp4 significantly at 6 hours and 24 hours. IQ1 increased the
expression of Sox2 and Scgb1a1. Explants groups: 6 hours treatment (0 h, n = 8, DMSO n = 8, IQ1, n = 9, ICG-001, n = 3), 24 hours treatment
(0 h, n = 6, DMSO, n = 6, IQ1, n = 6, ICG-001, n = 6). Data are presented as mean ± SD. *p < 0.05 vs. control by t-test. (B) Whole mount in situ
hybridization shows local expression of Bmp4 and Nkx2-1 in the distal epithelium. IQ1 decreased the expression of Bmp4 and Nkx2-1 at 24 hours. Figure 4 Gene expression analyses of IQ1 treated lung explants. (A) Quantitative PCR (qPCR) analysis of the lung explants. Lung explants
were cultured with either DMSO (control), IQ1 (10 μM) or ICG-001 (10 μM) for 6 hours or 24 hours. Relative expression changes compared to
E12.5 lungs without culture (0 h) are presented. IQ1 decreased the expression of Bmp4 significantly at 6 hours and 24 hours. IQ1 increased the
expression of Sox2 and Scgb1a1. Explants groups: 6 hours treatment (0 h, n = 8, DMSO n = 8, IQ1, n = 9, ICG-001, n = 3), 24 hours treatment
(0 h, n = 6, DMSO, n = 6, IQ1, n = 6, ICG-001, n = 6). Data are presented as mean ± SD. *p < 0.05 vs. control by t-test. Inhibition of the β-catenin/p300 interaction and branching
morphogenesis by IQ1 is reversible Finally, we addressed the question of whether or not
inhibition of branching morphogenesis by IQ1 disrup-
tion of the β-catenin/p300 interaction is reversible. To
examine this question, we first cultured lung explants
with IQ1 for 48 hours and either maintained the treat-
ment with IQ1 or removed the IQ1 from the culture
media. After an additional 48 hours of ex vivo culture
conditions without IQ1, new branching at the distal
tips was observed, whereas in the presence of IQ1, dis-
tal branching was still absent as anticipated (Figure 5A). Similarly, we treated E12.5 mouse embryos in utero for
one day and then stopped IQ1 feeding to the pregnant
mothers for an additional two days. In the lungs transi-
ently treated with IQ1, we found distal branching tips,
essentially equivalent to the untreated controls, although
the overall lung lobe was smaller, presumably due to the
temporary delay in development (Figure 5B). By immuno-
histochemistry of alpha smooth muscle actin (α-SMA),
a marker of myofibroblasts, we observed myofibroblasts Discussion Wnt/β-catenin signaling is critical during multiple stages
of lung development [5]. Based upon genetically modified
mouse models, Wnt/β-catenin signaling has also been im-
plicated in the regulation of the proximal-distal axis during
embryonic lung development. We now provide evidence
that interaction between β-catenin and specifically the Kat3
coactivator p300 plays a critical role in proximal-distal axis
determination during epithelial lung branching morpho-
genesis. Inhibition of the β-catenin/p300 interaction, using
the previously characterized small molecule inhibitor IQ1
[7] disrupted distal branching of murine lung epithelium in
utero (Figure 2A) and ex vivo (Figure 2D). The phenotype
of the IQ1 treated lungs involved proximalization as judged
by decreased expression of the distal genes Bmp4, Nkx2-1
(Ttf-1) and Fgf10 and increased expression of the proximal
genes and Sox2 and Scgb1a1 (CC10) (Figure 4A). Bmp4
and Nkx2-1 are expressed in the distal epithelium, Figure 5 Branching restarts on removal of IQ1. (A) E13.5 lung explants were cultured with IQ1 for 48 hours. Subsequently, the IQ1 was
removed from the culture media. After two days of the additional culture without IQ1, new branching ends were found at the distal tips (arrows). (B) E11.5 embryos were treated with IQ1 in utero for one day. The embryos were subsequently dissected at E14.5 after two days of growth in the
absence of IQ1. ALP staining of the left lobe shows fine branching equivalent to the control although the lobe is smaller. (C) E13.5 lung explants
were cultured with either DMSO (control) or IQ1 (10 μM) for 24 hours. Immunohistochemistry of α-SMA shows myofibroblasts surrounding the
proximal airway. However, myofibroblasts are absent at the distal tips in both the control and IQ1 treated lung explants (arrows). Nuclei were
stained with hematoxylin. Figure 5 Branching restarts on removal of IQ1. (A) E13.5 lung explants were cultured with IQ1 for 48 hours. Subsequently, the IQ1 was
removed from the culture media. After two days of the additional culture without IQ1, new branching ends were found at the distal tips (arrows). (B) E11.5 embryos were treated with IQ1 in utero for one day. The embryos were subsequently dissected at E14.5 after two days of growth in the
absence of IQ1. ALP staining of the left lobe shows fine branching equivalent to the control although the lobe is smaller. (C) E13.5 lung explants
were cultured with either DMSO (control) or IQ1 (10 μM) for 24 hours. Proximalizations of the embryonic lung selective via
inhibition of the β-catenin/p300 interaction or β-catenin
deletion are mechanistically distinct β-catenin is a multifunctional protein that beyond its
role as a transcriptional activator of Wnt signaling, is also
an important protein component of adherens junction,
which are important for epithelial cell-cell adhesion. To test
if the inhibition of the β-catenin/p300 interaction affected
the expression and localization of β-catenin in the lung
epithelium, we next examined β-catenin by immunohis-
tochemistry. We found β-catenin widely expressed in
both the epithelium and mesenchyme of the control treated
lungs (Figure 3B). Localization of β-catenin at the epithelial
cell membrane (Figure 3B, arrows) is consistent with its
important role in cell-cell adhesion in the lung epithelium. However, inhibition of the β-catenin/p300 interaction
by IQ1 did not change the expression or localization of
β-catenin whereas deletion of β-catenin completely
eliminates it. Genetic deletion of β-catenin also causes a disruption
in angiogenesis of peripheral vessels [6]. To examine if
inhibition of the β-catenin/p300 interaction disrupted
angiogenesis of peripheral vessels, we immunostained
whole mount lungs with an antibody specific for the
endothelial cell marker PECAM-1. Compared to the vehicle Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
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http://www.transrespmed.com/content/2/1/8 Page 7 of 10 surrounding the proximal airway but not the distal epi-
thelial tips in both the control and IQ1 treated lungs
(Figure 5C). The absence of myofibroblasts at the distal
tips of the IQ1 treated lungs implies that the distal tips
maintain their distal properties and maintain the plasticity
to undergo branching morphogenesis upon removal of
the inhibitory effects of IQ1. We further speculate that
this plasticity, under continuous IQ1 treatment fosters
the elongation of the epithelial tubes without branching
(Figure 2A). These results suggest that β-catenin/p300
inhibition by IQ1 transiently disrupts lung branching
morphogenesis but maintains the plasticity of the epithelial
cells at the distal tips for at least 24 hours. treated lungs, we found extensive PECAM-1 staining in the
capillary endothelium surrounding the distal tips of the IQ1
treated lungs (Figure 3C). This result suggests that IQ1 did
not dramatically disrupt angiogenesis of peripheral capillary
vessels whereas genetic deletion of β-catenin does. Based on these experiments, we suggest that although
phenotypically similar, the inhibitions of branching
morphogenesis via selective pharmacological inhibition
of the β-catenin/p300 interaction or genetic deletion of
β-catenin are mechanistically distinct. Discussion Immunohistochemistry of α-SMA shows myofibroblasts surrounding the
proximal airway. However, myofibroblasts are absent at the distal tips in both the control and IQ1 treated lung explants (arrows). Nuclei were
stained with hematoxylin. Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Page 8 of 10 whereas Fgf10 is expressed in the distal mesenchyme [1]. On the other hand, Sox2 is expressed in the proximal
epithelium [15] and Scgb1a1 is a proximal secretory
cell marker, expressed primarily in Clara cells [16]. This
proximalization that is induced by specific inhibition
of β-catenin/p300 dependent transcription extends our
understanding and is fully consistent with previous reports
of genetic deletion of β-catenin in the lung epithelium
and inhibition of Wnt signaling via Dkk-1 overexpres-
sion inhibiting branching lung morphogenesis [6,14,17]. These earlier investigations in conjunction with our
current study, confirm that β-catenin and in particular
its interaction with the transcriptional coactivator p300
is critical for proximal-distal determination during lung
branching morphogenesis. Our chemical genomic approach
that selectively inhibits only a subset of the transcriptional
roles of β-catenin, without interfering with its role in adhe-
rens complexes at the cell membrane, provides convincing
evidence that transcriptional function of β-catenin, not
the cell-cell adhesion function is critical for proximal-
distal determination. Interestingly, Shikama et al. had
previously demonstrated that embryonic mice with even a
single allele carrying a defective p300 Histone acetyltrans-
ferase (HAT) domain die at birth due to respiratory failure,
whereas CBP HAT defective embryos did not demonstrate
major lung developmental defects [18]. This is consistent
with our results that demonstrate that specific disrup-
tion of the β-catenin/p300 dramatically inhibits branch-
ing
lung
morphogenesis,
although
the
phenotypes
induced by IQ1 specifically blocking the β-catenin/p300
interaction and knock-in of defective p300 HAT activity
do diverge in other respects. distal lung mesenchyme in an indirect fashion [24]. Our
qPCR and in situ hybridization data suggest that expression
of both Bmp4 and Fgf10 is dependent on β-catenin/p300
transcription. Consistent with our results, both Bmp4
and Fgf10 have been previously identified as downstream
Wnt/β-catenin target genes during proximal-distal pattern-
ing in the lung [14,25]. Our result correspondingly indicates
that β-catenin/p300 transcription regulates the expression
of Bmp4 and Fgf10 during lung development. Interestingly,
inhibition of the corresponding β-catenin/CBP inter-
action with ICG-001 did not disrupt branching signifi-
cantly ex vivo (Figure 2E and 3A) and did not affect
normal lung development in utero (Figure 2B). Discussion The sig-
nificant differences between the effects of the specific
small molecule coactivator modulators IQ1 and ICG-001
further demonstrates that Wnt/β-catenin regulated gene
transcription is dramatically dependent on its choice
amongst the two coactivators, p300 and CBP and that
the β-catenin/p300 interaction is essential during lung
branching morphogenesis. This result is again fully con-
sistent with the previous results of Shikama et al. [18]. p
Activation of the Wnt/β-catenin signaling cascade is
critical during lung development and has been impli-
cated in the restoration of normal tissue structure and
function, as well as remodeling/fibrosis in a number of
organs, including the lung, suggesting that this devel-
opmental pathway can be reactivated in adult tissues
following injury [26,27]. However, exquisite control over
reactivation of these developmental pathways is critical,
as aberrant activation is associated with lung disease in-
cluding asthma and fibrosis [28,29]. Therefore, a precise
understanding of the role of differential coactivator usage
in the Wnt/β-catenin pathway during development and
repair after injury is important. In particular, the ability
to safely therapeutically modulate coactivator usage has
a number of clinical ramifications [30]. We previously
demonstrated that administration of the CBP/β-catenin
antagonist ICG-001 prevents and reverses established
lung fibrosis and dramatically improves animal survival
[2]. Similar results have been demonstrated in experi-
mental models of kidney fibrosis [31], and acute lung
injury [32]. Impaired branching of the alveolar tree in
genetically modified mice leads to respiratory failure
resulting from decreased surface for gas exchange. Overall, the lung phenotype bears similarity to respira-
tory distress syndrome and bronchopulmonary dyspla-
sia, the most common complications of prematurity in
humans [33]. Based upon our previous results and the
results of our current study, we propose that CBP/catenin
antagonists could be safely utilized to treat respiratory
distress syndrome and bronchopulmonary dysplasia in
infants. In this regard, the recently developed second
generation specific Wnt/CBP/catenin antagonist PRI-
724 (IC50 ~ 150 nM) has proven extremely safe in both We propose that one of the primary mechanisms by
which inhibition of the β-catenin/p300 interaction in-
duces lung proximalization involves the downregula-
tion of Bmp4 and Fgf10 expression. Bmp4 and Fgf10
are both essential growth factors in lung branching
morphogenesis to induce the growth of distal tips [19]. Formation of the distal respiratory epithelium and
branching of the proximal airways requires BMP4, which
is expressed at the tip of elongating epithelial buds [20]. Conclusions 6. Mucenski ML, Wert SE, Nation JM, Loudy DE, Huelsken J, Birchmeier W,
Morrisey EE, Whitsett JA: beta-Catenin is required for specification of
proximal/distal cell fate during lung morphogenesis. J Biol Chem 2003,
278:40231–40238. Our study provides strong evidence that the interaction
between β-catenin and p300 is critically important in
proximal-distal axis determination during mouse lung
branching morphogenesis. Our results are consistent and
add further refinement to multiple previous reports regard-
ing the role of β-catenin on proximal-distal determination
in the lung based upon genetic deletion of β-catenin as well
as previous reports on the differential roles of the coactiva-
tors CBP and p300 during lung development. The chemical
genomic approach utilized herein further elucidated and
clarified the specific role of the coactivator p300 in the
regulation of Wnt/β-catenin driven branching morphogen-
esis in the lung. This information, along with the ability to
safely pharmacologically manipulate differential coactivator
usage in the Wnt/β-catenin cascade provides multiple
therapeutic opportunities to investigate going forward. 7. Miyabayashi T, Teo JL, Yamamoto M, McMillan M, Nguyen C, Kahn M: Wnt/
beta-catenin/CBP signaling maintains long-term murine embryonic stem
cell pluripotency. Proc Natl Acad Sci U S A 2007, 104:5668–5673. 7. Miyabayashi T, Teo JL, Yamamoto M, McMillan M, Nguyen C, Kahn M: Wnt/
beta-catenin/CBP signaling maintains long-term murine embryonic stem
cell pluripotency. Proc Natl Acad Sci U S A 2007, 104:5668–5673. 8. Emami KH, Nguyen C, Ma H, Kim DH, Jeong KW, Eguchi M, Moon RT, Teo
JL, Oh SW, Kim HY, Moon SH, Ha JR, Kahn M: A small molecule inhibitor of
beta-catenin/CREB-binding protein transcription [corrected]. Proc Natl
Acad Sci U S A 2004, 101:12682–12687. 9. Teo JL, Ma H, Nguyen C, Lam C, Kahn M: Specific inhibition of CBP/beta-
catenin interaction rescues defects in neuronal differentiation caused by
a presenilin-1 mutation. Proc Natl Acad Sci U S A 2005, 102:12171–12176. 10. Hasegawa K, Yasuda SY, Teo JL, Nguyen C, McMillan M, Hsieh CL, Suemori
H, Nakatsuji N, Yamamoto M, Miyabayashi T, Lutzko C, Pera MF, Kahn M:
Wnt signaling orchestration with a small molecule DYRK inhibitor
provides long-term xeno-free human pluripotent cell expansion. Stem Cells Transl Med 2012, 1:18–28. 11. Eguchi M, Nguyen C, Lee SC, Kahn M: ICG-001, a novel small molecule
regulator of TCF/beta-catenin transcription. Med Chem 2005, 1:467–472. 12. Discussion Inhibition of BMP signaling in utero by overexpression
of Noggin resulted in lung proximalization [20]. Fgf10 is
expressed in the lung mesenchyme and reductions in
Fgf10 expression result in compaction of terminal sac-
cules and neonatal lethality [21,22]. In our study, inhib-
ition of the β-catenin/p300 interaction decreased the
expression of both Bmp4 and Fgf10 in the lung explants
(Figure 4A). The downregulation of Bmp4 expression
was significant already at 6 hours after IQ1 treatment,
consistent with BMP4 being a direct Wnt/β-catenin
target gene [23]. FGF10 expression although essentially
unchanged at 6 hours, was significantly decreased at
24 hours consistent with it being Wnt regulated in the Page 9 of 10 Page 9 of 10 Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Investigational New Drug (IND) enabling toxicology
studies and in a Phase Ia clinical trial [34]. In regards to re-
generative medicine, ex vivo small molecule modulation of
stem and progenitors cells offers another avenue to pursue. In that regard, Banerjee et al. demonstrated that human
embryonic stem cells (hES cells) directed to alveolar type II
(ATII) lung progenitor cells, could be differentiated into
an alveolar type I (ATI) phenotype following incubation
with the CBP/β-catenin antagonist ICG-001 [35]. One
week after acute lung injury induced by bleomycin ad-
ministration, these differentiated hES cells were able to
home to the small airways and engraft, with an accompan-
ied marked reduction in collagen deposition and tissue
damage. In the future, the potential to construct bioartificial
lungs could provide a source for organs for transplantation. Bioartificial lungs would provide a mechanism to circum-
vent the problems of donor shortage and immunogenicity. Unlike decellularized lung, synthetic bioartificial scaffolds
lack the endogenous growth factors that are required to
signal the seeded progenitor cells to essentially recapitu-
late ex vivo the lung development process [36]. Small
molecule manipulation of signaling pathways may be
used to guide proximal–distal patterning of lung pro-
genitor cells seeded on a synthetic bioartificial scaffold. Competing interests Competing interests
MK is a consultant, and equity holder in Prism Pharma, which is developing
the CBP/β-catenin antagonist PRI-724. MK is a consultant, and equity holder in Prism Pharma, which is developing
the CBP/β-catenin antagonist PRI-724. MK is a consultant, and equity holder in Prism Pharma, which is developing
the CBP/β-catenin antagonist PRI-724. p
p
g
g
15. Que J, Luo X, Schwartz RJ, Hogan BL: Multiple roles for Sox2 in the
developing and adult mouse trachea. Development 2009, 136:1899–1907. Abbreviations
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tetrazolium chloride; BCIP: 5-bromo-4-chloro-3’-indolyphosphate p-toluidine
salt; qPCR: Quantitative polymerase chain reaction; ALP: Alkaline
phosphatase; HAT: Histone acetyltransferase; hES cells: Human embryonic
stem cells. 13. Sasaki T, Hwang H, Nguyen C, Kloner RA, Kahn M: The small molecule Wnt
signaling modulator ICG-001 improves contractile function in chronically
infarcted rat myocardium. PLoS One 2013, 8:e75010. infarcted rat myocardium. PLoS One 2013, 8:e75010. 14. Shu WG, Guttentag S, Wang ZS, Andl T, Ballard P, Lu MM, Piccolo S,
Birchmeier W, Whitsett JA, Millar SE, Morrisey EE: Wnt/beta-catenin
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developing and adult mouse trachea. Development 2009, 136:1899–1907. 16. Hashimoto S, Nakano H, Singh G, Katyal S: Expression of Spred and
Sprouty in developing rat lung. Gene Expr Patterns 2002, 2:347–353. 17. De Langhe SP, Sala FG, Del Moral PM, Fairbanks TJ, Yamada KM, Warburton
D, Burns RC, Bellusci S: Dickkopf-1 (DKK1) reveals that fibronectin is a 14. Shu WG, Guttentag S, Wang ZS, Andl T, Ballard P, Lu MM, Piccolo S,
Birchmeier W, Whitsett JA, Millar SE, Morrisey EE: Wnt/beta-catenin
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Th
d This study was supported by funding from NIH 1R01 HL112638-01 and funds
from the University of Southern California. We thank Dr. Savério Bellusci for
kind comments. Author details
1 1Department of Biochemistry and Molecular Biology, Keck School of
Medicine, University of Southern California, 1450 Biggy Street, Los Angeles,
CA 90033, USA. 2Center for Molecular Pathways and Drug Discovery, Keck
School of Medicine, University of Southern California, 1450 Biggy Street, Los
Angeles, CA 90033, USA. 3Norris Comprehensive Cancer Center, Keck School
of Medicine, University of Southern California, 1450 Biggy Street, Los Angeles,
CA 90033, USA. Received: 14 May 2014 Accepted: 1 August 2014 Received: 14 May 2014 Accepted: 1 August 2014 Received: 14 May 2014 Accepted: 1 August 2014 Conclusions Schenke-Layland K, Nsair A, Van Handel B, Angelis E, Gluck JM, Votteler M,
Goldhaber JI, Mikkola HK, Kahn M, Maclellan WR: Recapitulation of the
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phosphatase; HAT: Histone acetyltransferase; hES cells: Human embryonic
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mouse embryonic lung. Dev Biol 2005, 277:316–331. major target of Wnt signaling in branching morphogenesis of the
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Oncogenic beta-catenin is required for bone morphogenetic protein 4
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Fgf10 expression in airway smooth muscle after airway epithelial injury
in mouse. PLoS One 2013, 8:e71426. 25. De Langhe SP, Carraro G, Tefft D, Li C, Xu X, Chai Y, Minoo P, Hajihosseini
MK, Drouin J, Kaartinen V, Bellusci S: Formation and differentiation of
multiple mesenchymal lineages during lung development is regulated
by beta-catenin signaling. PLoS One 2008, 3:e1516. 26. Königshoff M, Eickelberg O: WNT signaling in lung disease: a failure or a
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pathway for tissue repair. Trends Biotechnol 2009, 27:131–136. 28. Roth HM, Wadsworth SJ, Kahn M, Knight DA: The airway epithelium
in asthma: developmental issues that scar the airways for life? Pulm Pharmacol Ther 2012, 25:420–426. 29. Chilosi M, Poletti V, Zamò A, Lestani M, Montagna L, Piccoli P, Pedron S,
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Doglioni C: Aberrant Wnt/beta-catenin pathway activation in idiopathic
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TS
d
h 16. Hashimoto S, Nakano H, Singh G, Katyal S: Expression of Spred and
Sprouty in developing rat lung. Gene Expr Patterns 2002, 2:347–353. TS carried out the experiments. TS and MK designed the experiments and
drafted the manuscript. Both authors read and approved the final
manuscript. TS carried out the experiments. TS and MK designed the experiments and
drafted the manuscript. Both authors read and approved the final
manuscript. 17. De Langhe SP, Sala FG, Del Moral PM, Fairbanks TJ, Yamada KM, Warburton
D, Burns RC, Bellusci S: Dickkopf-1 (DKK1) reveals that fibronectin is a 17. De Langhe SP, Sala FG, Del Moral PM, Fairbanks TJ, Yamada KM, Warburton
D, Burns RC, Bellusci S: Dickkopf-1 (DKK1) reveals that fibronectin is a Page 10 of 10 Page 10 of 10 Sasaki and Kahn Translational Respiratory Medicine 2014, 2:8
http://www.transrespmed.com/content/2/1/8 Gottardi CJ, Königshoff M: Considerations for targeting β-catenin signaling
in fibrosis. Am J Respir Crit Care Med 2013, 187:566–568. 31. Hao S, He W, Li Y, Ding H, Hou Y, Nie J, Hou FF, Kahn M, Liu Y: Targeted
inhibition of β-catenin/CBP signaling ameliorates renal interstitial
fibrosis. J Am Soc Nephrol 2011, 22:1642–1653. 32. Zemans RL, Briones N, Campbell M, McClendon J, Young SK, Suzuki T, Yang
IV, De Langhe S, Reynolds SD, Mason RJ, Kahn M, Henson PM, Colgan SP,
Downey GP: Neutrophil transmigration triggers repair of the lung
epithelium via beta-catenin signaling. Proc Natl Acad Sci U S A 2011,
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applications to clinical practice? Ann Med 1997, 29:531–534. 34. El-Khoueiry AB, Ning Y, Yang D, Cole S, Kahn M, Zoghbi M, Berg J, Fujimori
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Henderson WR: Human embryonic stem cells differentiated to lung
lineage-specific cells ameliorate pulmonary fibrosis in a xenograft
transplant mouse model. PLoS One 2012, 7:e33165. Submit your manuscript to a
journal and benefi t from:
7 Convenient online submission
7 Rigorous peer review
7 Immediate publication on acceptance
7 Open access: articles freely available online
7 High visibility within the fi eld
7 Retaining the copyright to your article
Submit your next manuscript at 7 springeropen.com Submit your manuscript to a
journal and benefi t from:
7 Convenient online submission
7 Rigorous peer review
7 Immediate publication on acceptance
7 Open access: articles freely available online
7 High visibility within the fi eld
7 Retaining the copyright to your article
Submit your next manuscript at 7 springeropen.com 36. Lemon G, Lim ML, Ajalloueian F, Macchiarini P: The development of the
bioartificial lung. Br Med Bull 2014, 110:35–45. doi:10.1186/s40247-014-0008-1
Cite this article as: Sasaki and Kahn: Inhibition of β-catenin/p300 interaction
proximalizes mouse embryonic lung epithelium. Translational Respiratory
Medicine 2014 2:8.
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Deep state-space modeling for explainable representation, analysis, and forecasting of professional human body dynamics in dexterity understanding and computational ergonomics. Robotics [cs.RO]. Université Paris sciences et lettres, 2023. English. ⟨NNT : 2023UPSLM025⟩. ⟨tel-04250527⟩
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The percentage of robot spatial adaptation (SA) and reduction in operator’s movement (RiOM) [Papanagiotou, 2021; Olivas-Padilla, 2023] were used as key performance indicators (KPI). The KPI of robot spatial adaptation represents the ratio of the distance covered by the robot without spatial adaptation to the distance covered when the robot adjusts to the operator-specified position. The following formula is used to determine this KPI: SA(%) = ∥AHP − WP∥ − ∥PHP − WP∥ ∥PHP − WP∥ (6.7) where SA is spatial adaptation, AHP the adapted handover position, WP is the waiting point and PHP the particular handover position. Centimeters are used to measure distances. The higher the rate of adaptation, the more effort the operator had to put in during the HRC 127 Chapter 6. Computational ergonomics for task delegation in HRC scenario without the spatial adaptation of the robot. RiOM quantifies the difference in operators’ movement before and after introducing gesture recognition. This KPI is calculated as follows: RiOM(%) = ∥MwoGR∥ − ∥MwGR∥ ∥MwoGR∥ (6.8) MwoGR corresponds to the movement without gesture recognition, and MwGR is the movement with gesture recognition. This KPI measures the amount of effort reduced by the operator as a result of gesture recognition. SA is primarily used to compare the first experiment (physical interaction) with the third experiment (spatial adaptation with pose estimation), whereas RiOM is used to compare the first experiment with the second experiment (temporal adaptation with gesture recognition).
6.6.1.2 Results and discussion
The 3DCNNs were trained with the command gesture dataset for the gesture recognition module and demonstrated 98.50% accuracy in recognizing the 11 gestures. All the operators completed the collaboration procedure successfully, indicating that the accuracy of the recognition algorithm is sufficient even for users that were not part of the training dataset. The calculated KPIs for each operator are shown in Table 6.3. Note that the greater the percentage of SA, the more difficult it was for the operator to receive the cards without spatial adaptation enabled. This is because the predefined handover position was not close to where the operator would prefer to receive the cards. For RiOM, however, the larger the percentage, the better, as it indicates a greater reduction in operator’s movement when gesture recognition is utilized. The average rate of spatial adaptation and reduction in operator’s movement were 29.37% and 28.37%, respectively, among the 14 subjects. Compared to the configuration without gesture recognition, the optimized HRC configuration significantly reduced the operators’ movement. This is also demonstrated by seeing Figure 6.6a, where the operator was required to rotate his torso in order to touch the robot’s sensor, which is located outside the TV chassis. In Figure 6.6b, the robot recognizes when the operator has completed his task and can proceed to the next action in the work routine. As demonstrated by the KPIs, the proposed HRC scenario enhances ergonomics and efficiency. This is accomplished by first assigning the hazardous tasks from the original scenario to the collaborative robot. Then, integrating gesture recognition and spatial adaptation to prevent operators from performing unnecessary movements that could cause physical discomfort.
Table 6.3:
Measured
KPI
s for each operator. KPI SA(%) RiOM(%) 1 39.10 31.40 2 33.30 33.10 3 21.10 24.40 4 27.50 27.10 5 30.40 32.10 6 31.90 27.30 Operator 7 8 27.10 31.80 24.50 26.80 128 9 13.40 37.40 10 33.90 20.60 11 43.50 45.90 12 32.10 20.80 13 18.70 21.30 14 27.40 24.50
6.7
.
Conclusion of the chapter
(
a
)
MwoGR: Press robot to start (b) MwGR: The robot recognizes G7 to start
Figure 6.6: TV assembly with and without gesture recognition.
6.7 Conclusion of the chapter
This chapter presents a methodology for designing a human-robot collaboration framework that maximizes ergonomics and production efficiency in a television co-production cell. The first step was creating a system for identifying four postural risk factors. According to the risk factors detected, an ergonomic risk score is calculated based on EAWS. This system was trained using the 28 motion primitives from the ERGD dataset. The trained system successfully recognized the four risk factors using only data from a minimal set of sensors, selected in Chapter 5. Following the training of the automatic ergonomic scoring system, real professional tasks performed on a television production line were evaluated for task delegation. Two of the four evaluated tasks were assigned to a collaborative robot. Hence, the suggested HRC framework for this scenario consists of the robot grabbing the circuit board and wire from their respective containers and handing them to the human operator. The human operator then connects the wire to the circuit board, positions the board and wire on the television chassis, then drills the board into the chassis. This analysis can be applied to other professional tasks for rapid reconfigurability, just as it was done with TV assembly tasks. First, the professional tasks to be evaluated need to be recorded with the designated inertial sensors and placed according to the standards of the ISB. Next, calculate the Euler angles of each body part measured by each sensor. Then, segment the data of the tasks into four-second windows, ideally with a two-second overlap to cover the entire task. Finally, apply the automatic postural evaluation to the segmented tasks, which would indicate the motion primitives detected and the estimated EAWS score. Depending on the nature of the identified high-risk tasks, it is determined whether they should be delegated to a collaborative robot or the production cell should be redesigned to prevent the performance of hazardous movements. Thus, HRC frameworks can be ergonomically improved by implementing the task delegation process presented in this chapter and the HRC design proposed by Papanagiotou et al. [Papanagiotou, 2021; Olivas-Padilla, 2023]. First, it is identified which risky tasks should be assigned to the collaborative robot. Then, by applying gesture recognition and spatial adaptation, the robot can assist operators in reducing their range of motion so that the operators perform only safe and convenient movements. As a result, less physical effort is required from them to fulfill their professional duties. Chapter 6. Computational ergonomics for task delegation in HRC
Lastly, it is worth noting that wearables that measure working postures have the potential to decrease the incidence of WMSDs. High-frequency and easily-accessible monitoring technology can provide feedback to managers and operators on how to address exposures to ergonomic risks. In this regard, Appendix B describes automatic ergonomic evaluation applications designed to utilize MoCap data for ergonomically analyzing human movements. Future plans for this platform include implementing the proposed automatic ergonomic evaluation in this chapter and the human movement analysis methods presented so that users can apply them to analyze their own recorded movements or learn from those in Chapter 3.
Conclusions 7.1 Summary
This dissertation was primarily focused on creating methodologies for training interpretable human motion models utilizing practical and portable MoCap technologies, such as IMUs. These models could be used to generate accurate human movements and gain insight into the dynamics of human movement during the execution of a movement. The applicability of statespace models for developing a generalized motion understanding framework was investigated. Consequently, three approaches that adhere to the structure of the Gesture Operational Model were proposed. The fourth chapter presented ideas for combining state-space models and data-driven approaches to train interpretable GOM representations of human movements that enable the simulation of accurate 3D human postures. The proposed methods were able to parameterize the conditional distributions specified in the state-space models. The generated models exhibited their potential to learn human movements in a general and scalable way, as they were able to fit data distributions from reduced data sets and recorded with different subjects in different scenarios. The difference in performance between the three approaches may be influenced by their structure and number of parameters. Additional research is required to get a complete understanding of how these two factors interact. The use of either a statistical or data-driven approach for human motion representation would depend on the nature of the motion-based application. For instance, whether a single or several human movements are examined, as well as the processing power constraints. The proposed approaches can be simply implemented utilizing existing statistical and deep learning libraries. Additionally, they can be upgraded with new advancements in deep learning or incorporated into more sophisticated architectures, such as Generative Adversarial Networks (GANs) [Yoon, 2019], that compute the gradients of any differentiable architecture using automatic differentiation techniques. As described in Chapter 5, the trained models allowed the body dexterity analysis of industrial operators and skilled craftsmen. This analysis described how body joints collaborate to accomplish specified motion trajectories. With the motion representations, it was also possible to perform a selection of meaningful motion descriptors for modeling a set of human movements. This selection method could be utilized for a broad range of applications requiring the modeling of a specific set of movements using a minimal sensor configuration. Conclusions
Chapter 6 presented an application of the selection of meaningful motion descriptors while creating a methodology for automatic ergonomic analysis and task delegation in HRC frameworks. Professional tasks were assessed by first recording them with a minimal set of selected sensors. Afterward, an EAWS-based ergonomic score was automatically calculated based on the detected motion patterns. The task delegation then consisted of assigning the tasks with the most dangerous movements to the collaborative robot, while the operators were allocated the ergonomically safe or supervisory control tasks. The main scientific and technological contributions of this thesis are listed next.
7.1.1 Scientific contributions Three methods for learning of interpretable human motion models
The methods generate interpretable time-varying models of human movements to study body dexterity and create realistic motion simulations. Human movements are represented using GOM, which consists of a set of autoregressive models, each modeling a different joint motion descriptor (joint angle). The first method estimates the model’s parameters using Kalman filters (KF-RGOM) with one-shot training. The second and third methods correspond to deep state-space models. The second method utilizes a stochastic autoencoder (VAE-RGOM) to train multiple interpretable human motion representations, while the third method utilizes an autoencoder with the Luong attention mechanism (ATT-RGOM). KF-RGOM can be utilized for analyses where only small sets of human movements are available, as well as for applications that require only a few motion descriptors to be modeled. VAE-RGOM and ATT-RGOM are proposed for applications that need the analysis of multiple motion descriptors and human movements. Additionally, these methods can be used to augment data in deep learning applications. Analysis of full-body dexterity in industrial operators and expert artisans
A methodology based on trained motion representations is provided for analyzing the body dexterity of industrial operators and skilled artisans. This comprises a statistical analysis of the trained GOM representations to determine the significance of their assumptions in modeling a specific human movement. The results highlighted the key motion descriptors associated with and contributing to the whole-body movement, providing insights into how body joints collaborate to accomplish the predicted motion trajectories. In addition, identifying the most significant motion descriptors of all human movements associated with a professional task reveals the optimal set of motion descriptors for modeling and recognizing them. Finally, a procedure for computing tolerance intervals based on experts’ motion representations is provided to supplement the dexterity analysis. These tolerance intervals, which specify the acceptable range of motion for replicating a specific movement, are calculated using the parameters of multiple motion representations trained with different repetitions of the same movement.
7.1.2 Industrial and technological contributions Motion capture benchmark of industrial tasks and European historic crafts
A motion capture benchmark featuring datasets containing the full-body movements of real industrial operators and skilled craftsmen was developed. Currently, the most used datasets consist of common daily human movements. Therefore, seven new MoCap datasets of actual professional tasks performed in industry and crafts were created using an inertial based full-body MoCap suit. Methodology for improving HRC through computational ergonomics
A methodology for task delegation is developed in order to design the optimal HRC framework that maximizes ergonomics and production efficiency in a television co-production cell. Professional tasks done by human operators on the television production line are initially evaluated by recognizing postural risk factors using HMMs. According to the European Assembly Worksheet, an ergonomic risk score is calculated based on the detected risk factors. The optimal HRC configuration is then defined by delegating to the collaborative robot the hazardous tasks. Finally, the HRC scenario is enhanced by applying gesture recognition and spatial adaptation, which allow the human operator to collaborate with the robot using gestures while avoiding unnecessary movements that could cause physical strain.
7.2 Open questions and perspectives
This section concludes the dissertation by discussing the open questions and ideas regarding how the presented work could be expanded. Although the results obtained in this thesis already look promising, there is still room for improvements in the proposed frameworks and follow-up work on their implementation in real-world scenarios. Human motion representation with GOM
The GOM’s representation of human movement and the proposed estimation approaches can be further optimized. One of the advantages of GOM is its ability to easily incorporate new assumptions into its representations of human movement. In this dissertation, the mathematical model of human movements was based on kinematic measures of the body joints. Thus, it remains an open question if the accuracy of motion modeling can be improved by incorporating other types of measures, such as kinetic motion descriptors (joint torques or external forces), into the representation. Diverse applications, particularly in ergonomics, could benefit from the ability to get insight into how kinetic and kinematic measures interact to accomplish a specific movement. Previous studies have calculated joint torques for identifying the joints that accumulate the most strain during a variety of tasks [Menychtas, 2020]. Consequently, GOM representations with kinetic and kinematic measures could be utilized to create novel ergonomic monitoring systems for recognizing potential posture risks. For instance, the sensitivity analysis performed in Section 4.5 demonstrates the potential for using the estimated motion representations 133 Conclusions to analyze anomalous motion descriptor behavior. When performing an ergonomic analysis, it may be helpful to examine how the models react to shocks applied to various joint motion descriptors in order to later identify any physical strains (such as on the shoulders or lower back) or loads that may be affecting the workers’ performance during their shift. The professional tasks can then be modified to reduce the danger of injury. Parametrization of state-space models
The selection of the best architecture for parameterizing the GOM representations utilized in this thesis was not a trivial task, and there may be better default settings than those offered in this thesis. For example, the autoregressive order of the GOM models, the number of layers, units, or activation functions. In this dissertation, standard optimization algorithms were applied to determine the optimal settings for each architecture based on the dataset utilized. However, in order to avoid prolonged run times, the search for the best hyperparameter values was restricted to a specific range. As always, when using data-driven approaches, training tricks can make a huge difference in terms of the final performances of a model. Some were used in the training of VAE-RGOM and ATT-RGOM, but it would be interesting to find even more effective ones based on a deeper theoretical understanding of the learning process.
Inertial sensors for human motion capturing
Working with inertial-based MoCap data requires awareness of the limitations of using inertial sensors in real-world work environments. Inertial sensors can offer precise and reliable measurements to study human movement; however, the degree of this precision and reliability depends on the site, movements, and tools handled during the performance. In the recording for datasets GLB and APA, for example, subjects used plastic gloves or did not wear the gloves that come with the inertial suit to prevent measurement disturbances. Therefore, for implementing motion-based applications with inertial sensors, it is necessary to account for the possibility of magnetic disturbances during the recording of new datasets and to apply post-processing techniques to eliminate drifts in the measures that may influence the results of the proposed approaches. Explainable AI
This thesis is a step toward the development of explainable AI (XAI) for human motion modeling [Hagras, 2018]. Humans can easily comprehend and analyze the actions in XAI. As discussed in Chapter 2, conventional data-driven approaches and other supervised methods, such as linear or logistic regressions, can be difficult to interpret for high-dimensional motion data. These approaches do not permit the interpretation of human movements, nor do they explain the logic behind the trajectory predictions of joint motion descriptors. The work done in this thesis offers a first approach for generating interpretable human motion representations that can be used for dexterity analysis and other applications that require describing different human movements using only kinematic descriptors. Implementing human-computer interfaces that automatically apply statistical analysis to learned motion models and highlight significant motion descriptors 134 7.2. Open questions and perspectives in an intuitive and meaningful way could be the focus of future work. The intention would be to ease the user’s implementation of the proposed approaches on their own recorded movements or their understanding of movements contained in the benchmark presented in Chapter 3. Additionally, through the computation of the tolerance intervals outlined in Section 5.5, applications that provide users with real-time feedback about their capability to replicate particular movements could be designed. These interactive applications have the potential to enable novices to learn gross and fine motor skills even in the absence of an instructor by comparing their motion data to that of an expert during the instruction process. Simulating the movement of multiple individuals
Another potential application of the proposed motion representations can be their implementation in more sophisticated methods that simulate the movements of multiple individuals. Simulating human movements with the proposed motion representations implies fully utilizing the information in the MoCap data to predict future postures. Consequently, these learned representations might serve as the foundation for simulation. For example, to simulate the movement of several individuals, methods often use complex hypothetical decision rules, which frequently fail to produce realistic movements [Patterson, 2008; Rudenko, 2020]. These might be replaced by simulating from the proposed representations to generate the expected spatial distribution of the population. Appendix A General simulation results with all seven datasets Tables A.1, A.2, and A.3 present the average Mean Absolute Error (MAE), Root Mean Squared Error (RMSE), and Theil Inequality Coefficient (U1 ), respectively, achieved with each dataset and approach. All movements were generated with the respective motion representation of their class, then the MAE, RMSE, and U1 were calculated between the generated movement and the original.
Tables
A.
4
to
A
.8 illustrate the
simulation performance
based
on MAE for
each movement
within the datasets. Table A.1: Average MAE for each dataset. Dataset TVA TVP APA GLB SLW MSC ERGD KF-GOM 16.830 (σ: 15.379) 7.947 (σ: 5.278) 3.312 (σ: 2.816) 19.211 (σ: 9.981) 14.267 (σ: 7.739) 23.313 (σ: 14.514) 13.461 (σ: 8.699) KF-RGOM 6.938 (σ: 0.459) 9.867 (σ: 5.592) 10.946 (σ: 0.664) 12.916 (σ: 3.665) 9.207 (σ: 3.307) 16.002 (σ: 5.887) 13.569 (σ: 4.931) 136 VAE-RGOM 0.093 (σ: 0.016) 0.213 (σ: 0.058) 0.091 (σ: 0.019) 0.119 (σ: 0.044) 0.115 (σ: 0.041) 0.247 (σ: 0.101) 0.095 (σ: 0.052) ATT-RGOM 0.191 (σ: 0.024) 0.398 (σ: 0.085) 0.203 (σ: 0.048) 0.220 (σ: 0.066) 0.246 (σ: 0.078) 0.457 (σ: 0.126) 0.198 (σ: 0.085) Table A.2: Average RMSE for each dataset. Dataset TVA TVP APA GLB SLW MSC ERGD KF-GOM 32.438 (σ: 27.247) 15.978 (σ: 5.614) 17.814 (σ: 17.652) 42.918 (σ: 22.873) 28.787 (σ: 13.616) 51.455 (σ: 19.791) 21.732 (σ: 13.926) KF-RGOM 14.903 (σ: 1.574) 20.937 (σ: 2.391) 19.127 (σ: 10.266) 29.097 (σ: 10.373) 22.868 (σ: 10.798) 36.828 (σ: 22.618) 15.126 (σ: 11.006) VAE-RGOM 0.962 (σ: 0.430) 3.231 (σ: 1.402) 0.885 (σ: 0.147) 2.049 (σ: 1.384) 0.467 (σ: 0.287) 3.103 (σ: 2.043) 1.134 (σ: 0.758) ATT-RGOM 1.126 (σ: 0.410) 3.339 (σ: 1.389) 1.034 (σ: 0.146) 2.204 (σ: 1.388) 0.721 (σ: 0.328) 3.311 (σ: 1.980) 1.279 (σ: 0.782) Table A.3: Average U1 for each dataset. Dataset TVA TVP APA GLB SLW MSC ERGD KF-GOM 0.427 (σ: 0.368) 0.195 (σ: 0.068) 0.310 (σ: 0.192) 0.540 (σ: 0.221) 0.390 (σ: 0.341) 0.586 (σ: 0.262) 0.394 (σ: 0.281) KF-RGOM 0.384 (σ: 0.057) 0.125 (σ: 0.073) 0.210 (σ: 0.101) 0.292 (σ: 0.123) 0.201 (σ: 0.102) 0.361 (σ: 0.099) 0.2742 (σ: 0.056) VAE-RGOM 0.015 (σ: 0.005) 0.019 (σ: 0.004) 0.009 (σ: 0.003) 0.026 (σ: 0.015) 0.043 (σ: 0.016) 0.024 (σ: 0.010) 0.010 (σ: 0.003) ATT-RGOM 0.023 (σ: 0.005) 0.025 (σ: 0.003) 0.016 (σ: 0.003) 0.028 (σ: 0.015) 0.048 (σ: 0.013) 0.030 (σ: 0.009) 0.015 (σ: 0.003)
Table A.4: Mean absolute angle errors for TVA and TVP.
Dataset TVA TVP Motion TVA1 TVA2 TVA3 TVA4 TVP1 TVP2 TVP3 TVP4 TVP5 TVP6 TVP7 TVP8 TVP9 KF-GOM 25.610 37.600 2.742 1.368 2.050 4.779 5.545 2.267 12.305 15.816 16.452 4.132 8.178 KF-RGOM 7.226 6.271 6.804 7.450 3.241 5.746 5.164 9.657 13.669 15.192 21.746 6.898 7.491 137 VAE-RGOM 0.106 0.103 0.066 0.096 0.130 0.201 0.179 0.125 0.239 0.233 0.301 0.222 0.288 ATT-RGOM 0.218 0.211 0.160 0.176 0.310 0.309 0.375 0.256 0.446 0.480 0.508 0.480 0.419
Appendix A. General simulation results with all seven datasets
Table A.5: Mean absolute angle errors for APA and MSC. Dataset APA MSC Motion APA1 APA2 APA3 MSC1 MSC2 MSC3 MSC4 MSC5 MSC6 MSC7 MSC8 MSC9 MSC10 MSC11 MSC12 MSC13 KF-GOM 1.550 7.286 1.100 2 12.283 34.171 24.116 25.538 31.718 2.197 20.070 26.040 47.524 48.210 5.860 23.202 KF-RGOM 10.531 10.423 11.884 7.880 15.033 28.742 12.710 16.444 10.472 8.836 15.076 16.920 20.863 23.696 20.343 11.009 VAE-RGOM 0.089 0.114 0.069 0.139 0.156 0.323 0.183 0.204 0.230 0.132 0.273 0.432 0.288 0.453 0.160 0.236 ATT-RGOM 0.186 0.269 0.155 0.341 0.342 0.557 0.370 0.389 0.458 0.314 0.455 0.677 0.489 0.732 0.343 0.472
Table A.6: Mean absolute angle errors for SLW. Dataset SLW Motion SLW1 SLW2,1 SLW2,2 SLW2,3 SLW2,4 SLW2,5 SLW3 SLW4,1,1 SLW4,1,2 SLW4,1,3 SLW4,2,1 SLW4,2,2 SLW4,2,3 SLW4,3,1 SLW4,3,2 SLW4,3,3 KF-GOM 8.841 7.411 7.104 10.217 3.097 3.120 10.666 15.427 13.489 30.356 15.635 25.050 18.471 14.747 18.740 25.895 KF-RGOM 10.820 8.097 8.985 11.100 5.117 5.886 18.936 10.960 8.516 12.912 10.108 5.884 6.064 7.718 8.947 7.264 138 VAE-RGOM 0.097 0.069 0.068 0.099 0.067 0.118 0.092 0.217 0.185 0.130 0.154 0.098 0.133 0.123 0.081 0.111 ATT-RGOM 0.121 0.164 0.139 0.246 0.151 0.303 0.270 0.406 0.344 0.268 0.339 0.206 0.272 0.276 0.184 0.246
Table A.7: Mean absolute angle errors for GLB.
Dataset GLB Motion GLB1 GLB2 GLB3 GLB4 GLB5 GLB6 GLB7 GLB8 GLB9 GLB10 GLB11 GLB12 GLB13 GLB14 GLB15 GLB16 GLB17 GLB18 KF-GOM 36.340 2.929 21.410 11.502 24.251 17.904 5.213 28.045 21.055 25.070 32.240 7.859 18.714 33.678 9.203 7.300 15.606 27.487 KF-RGOM 18.491 10.239 19.363 9.898 11.521 9.500 10.483 16.302 16.196 15.138 15.081 11.642 15.717 12.708 8.919 6.998 7.684 16.608 139 VAE-RGOM 0.128 0.088 0.135 0.073 0.081 0.108 0.092 0.091 0.088 0.126 0.163 0.167 0.248 0.100 0.176 0.072 0.097 0.113 ATT-RGOM 0.231 0.170 0.228 0.146 0.155 0.209 0.187 0.177 0.180 0.245 0.276 0.265 0.407 0.193 0.333 0.146 0.191 0.221
Appendix A. General simulation results with all seven datasets Table A.8: Mean absolute angle errors for ERGD. Motion ERGD1 ERGD2 ERGD3 ERGD4 ERGD5 ERGD6 ERGD7 ERGD8 ERGD9 ERGD10 ERGD11 ERGD12 ERGD13 ERGD14 ERGD15 ERGD16 ERGD17 ERGD18 ERGD19 ERGD20 ERGD21 ERGD22 ERGD23 ERGD24 ERGD25 ERGD26 ERGD27 ERGD28 KF-GOM 6.516 4.378 2.633 3.332 2.759 4.750 3.657 3.090 3.063 18.234 22.209 6.586 4.952 16.796 17.635 8.923 10.190 18.354 22.967 18.294 15.333 16.639 15.077 23.038 27.285 29.911 23.421 26.899 KF-RGOM 2.825 7.014 6.187 6.329 8.543 14.041 10.544 12.626 13.229 15.835 20.697 13.920 11.037 13.568 6.105 14.033 14.444 15.565 13.936 15.073 17.560 12.833 12.473 19.642 19.341 20.268 19.252 23.024 140 VAE-RGOM 0.020 0.034 0.025 0.027 0.038 0.062 0.048 0.053 0.073 0.087 0.124 0.073 0.082 0.091 0.067 0.093 0.112 0.123 0.137 0.148 0.077 0.255 0.103 0.114 0.143 0.151 0.123 0.180 ATT-RGOM 0.039 0.077 0.057 0.065 0.084 0.162 0.109 0.124 0.190 0.192 0.256 0.181 0.200 0.221 0.167 0.219 0.227 0.261 0.308 0.329 0.155 0.262 0.207 0.241 0.283 0.308 0.259 0.362
Appendix B Web-based and Android-based applications for automated ergonomic evaluation B.1 Introduction
Manual laborers in the industry sector are often subject to critical physical strain that leads to work-related musculoskeletal disorders. Lifting, poor posture, and repetitive movements are among the causes of these disorders. In order to prevent them, several rules and methods have been established to identify ergonomic risks that workers might be exposed to during their activities. However, the ergonomic assessment through these methods is not a trivial task, and a relevant degree of theoretical knowledge on the part of the analyst is necessary. Therefore, this appendix presents a web-based and an android-based application for automatic ergonomic evaluation using MoCap data. The proposed applications use segment rotations (or joint angles) acquired from IMUs for the assessment and provide as feedback RULA scores, color visualizations, and limb angles in a simple, intuitive and meaningful way. RULA is one of the most commonly used observational methods for assessing occupational risk factors for upper-extremity musculoskeletal disorders. By automatizing RULA, an interesting perspective for extracting posture analytics for ergonomic assessment is opened, as well as the inclusion of new features that may complement it.
B.2 Automatic ergonomic evaluation module
Both applications use a module that automatically computes RULA scores based on a skeleton constructed using MoCap data. In this first version, the MoCap data is retrieved from BVH files generated either by the Notch Interfaces1 or Nansense MoCap systems. The module’s design is divided into three steps. The first step is to extract the segment rotations per 1 Notch Interfaces Inc. website: https://wearnotch.com/
Appendix B. Web-based and Android-based applications for ergonomic evaluation
Figure B.1: General scheme of the proposed RULA evaluation module. frame from the BVH file. Next,
the
RULA score
is computed by
apply
ing
thresholds
to the joint
rotation
s, followed by the
generation
of
color maps depending on the obtained scores
. Lastly, visual feedback is produced for ergonomic analysis. The main visual feedback for each application consists of three sections. The first section is composed of colored annotations based on the scores and color maps computed in the previous step. The second section is the Skeleton Sketch, which displays color annotations and skeleton drawings of various frames. The third section is comprised of the animation of the human motion data. Figure B.1 depicts the overall structure of the created module, whose primary components are described in the following subsections.
B.2.1 RULA computation
RULA is used to evaluate workers’ risk of developing upper extremity WMSDs. The evaluation considers posture, muscle use, and force applied during a task. According to RULA, the upper human body is divided into eight segments. Those segments are the trunk, the neck, two upper arms, two forearms, and two wrists. A score is assigned to each segment posture, as well as a score for exerted force and muscle activation [McAtamney, 1993]. As an example, Figure B.2 shows the thresholds defined in RULA for scoring the upper arm posture. The scores for the upper arms (SUPA ), neck (SN ), and trunk (ST ) can be from 1 to 6, the lower arms (SLA ) and wrists position (SWP ) from 1 to 4, and the legs (SL ) and wrist twist (SWT ) from 1 to 2. RULA has two other scores, Force score (SF ) and Muscle use score (SM ), where it considers external forces that the human might be exposed to and if the work posture is sustained for a long period or intermittently. After calculating all previous scores, the final RULA risk score (SRULA ) is computed from RULA’s Table C using the scores defined as score A (SA ) and score B (SB ) [McAtamney, 1993]. SA and SB can vary from 1 to 13. The SA is 142 B.2.
Automatic ergonomic evaluation module
Figure B.2: RULA scoring for the upper arm posture. Figure B.3: Location and Euler orientation of the joint angles provided by the Nansense system. obtained by the RULA’s Table A according to the scores SUPA, SLA, SWP, SWT, SF, and SM. SB is calculated from Table B using SN, ST, SL, SF, and SM [McAtamney, 1993]. SRULA can vary from 1 to 7. The highest score indicates a severe ergonomic risk, implying that the work posture must be changed immediately, and the lowest score a low risk, meaning that the work posture is acceptable and no change is needed. For automatically computing the RULA scores using MoCap data, joint angles sequences are extracted from BVH files. This initial version of the module utilizes BVH files generated from data recorded by IMUs. The BVH file format is split into two sections. The first section describes the skeleton’s hierarchy and initial posture. This section also lists the degrees of freedom and Euler orientation for each body part. The second section describes the channel data for each frame, which corresponds to the local joint angle sequences. For illustration purposes, the computation of the RULA scores using MoCap data recorded from the Nansense system is described next. Figure B.3 illustrates the joints measured and their Euler orientation in BVH files generated by the Nansense system. Next is explained the RULA score calculation that is applied to each motion data frame, with each frame representing a posture. Note that the ergonomist typically chooses one arm posture (left or right) in order to calculate the overall RULA score. Nonetheless, the module provides both scores, one based on the posture of the left arm and the other on the right arm. Also, due to the fact that RULA does not provide predefined thresholds for some scores, such as the wrist twist score, where the analyst must subjectively determine if the worker’s wrist is twisted or not, additional thresholds were introduced to achieve proper RULA scoring. These 143 Appendix B. Web-based and Android-based applications for ergonomic evaluation are detailed next, which consists of two score adjustments of the scores: SUPA, SN, and ST, and the threshold used to determine if the lower arms are twisted. The remaining thresholds are the same as the predefined by RULA [McAtamney, 1993]. SUPA is evaluated according to the rotation on the X-axis of the shoulders’ joints. In order to determine if the shoulder is raised, the rotation of the collars’ joints on the Z-axis (ZSH1 ) are examined. Since RULA does not define any angle threshold to determine if a shoulder is raised, the thresholds specified in Equation B.1 are used. Through these thresholds, SUPA is ′ modified, generating its adjusted version SUPA.
′ SUPA = SUPA + 1 if ZSH1 ≥ 10◦ SUPA if ZSH1 < 10◦ (B.1) The upper arm is indicated as abducted according to Equation B.2. The angle on the X-axis of the shoulder joint is defined as XSH2, the angle on the X-axis of the corresponding elbow ′′. (LFA or RFA) is XFA, and the final adjusted score is SUPA ′′ S
UPA
= SUPA + 1 if XSH2 ≥ 90◦ & XFA ≥ 5◦ ′ SUPA otherwise ′
(B.2) The angle on the X-axis of the elbow is used to calculate SLA, and the angle on the Z-axis of the same joint to identify whether the arm is moving across the body’s midline or outside. For SWP, the angles on the X and Z axes of the respective wrist joint are utilized to determine if the wrist is bent in a way that crosses the midline. SWT is calculated using the Y-axis angle of the corresponding elbow (YFA ). Since RULA does not establish thresholds for this situation, this score is defined as follows: SWT = 1
if −
45
◦ <
YFA
< 45
◦
2 otherwise (B.3) The neck flexion/extension, which corresponds to SN, is assessed by using the angle on the X-axis of the neck joint. To determine if the neck is twisted, the angle on the Y-axis (YN ) is used, where SN is adjusted (SN′ ) according to Equation B.4. SN′ = SN + 1 if YN ≥ 20◦ ||YN ≤ −20◦ SN otherwise (B.4) The angle on the
Z-axis of the neck joint
(
ZN
) is
utiliz
ed to
establish
w
hether or
not
the neck
is
flexed to a side;
the
threshold
defined
is
the following: SN′′ = SN + 1 if ZN ≥ 20◦ ||ZN ≤ −20◦ SN′ otherwise ′ (B.5) where SN′′ is the final RULA score of the neck region. ST is computed by analyzing the angle of the middle spine (SP2).
The
angle on the X-axis is
used
for measuring the bending, the Y-axis 144 B.2. Automatic ergonomic
evaluation
module (YSP2 ) to determine if
the trunk
is
twisted
,
and the Z-axis
(
ZSP
2 ) if there is
a
side
b
ending
. A
djustment
s
to ST for a twisted trunk
are
defined by Equation B.6 and for side
b
ending by
E
quation B
.
7
. ST + 1 if YSP2 ≥ 20◦ ||YSP2 ≤ −20◦ ST′ = (B.6) ST otherwise ST′′ = ST + 1 if ZSP2 ≥ 20◦ ||ZSP2 ≤ −20◦ ST′ otherwise ′ (B.7) where ST′ represents the first score adjustment and ST′′ is the final score for the trunk region. ′′, S ′′, and S ′′ are the scores used for these body regions in The final
adjusted
versions of SUPA N T
the
visual feedback and
final
RULA score calculation. B.2.2 Visual feedback for posture analytics B.2.2.1 Color mapping of RULA scores
Following the computation of all scores, a color mapping is performed. First, a color map is created for use in the mapping process. The number of possible values for a score is specified and denoted as n. For example, the score for the upper arm can vary from 1 to 6, so n = 6 for this score. The color maps are obtained as follows: C = [ch1, ch2, ch3 ] (B.8) Mn = [C1, C2,.., Cn ] (B.9) ch is between [0, 255] and Mn is an n-dimensional vector of RGB colors. The lower indexes in Mn are represented by green tones, the intermediate indexes by yellow tones, and the higher indexes by red tones. Next, the color mapping is done by using the following equation: CR = Mn [SR ] (B.10) ′′ ′′ ′′, S, S In Equation B.10, SR is a score of the RULA evaluation (SUPA LA WP, SWT, SN, ST, SL, SB, SA, or SRULA ), and CR is the color corresponding to SR. B.2.2.2 Graphical user interface
The main interfaces of the web-based and Android applications are shown in Figures B.4 and B.5, respectively. The applications were created using HTML, CSS, JavaScript, PHP, and Java. The Android app is compatible with operating systems 11 and up. The interface comprises four parts: the menu, the skeleton sketch, the human animation, and the score list. In the menu, it is possible to select the scores to display, configure the settings for score computation, and manipulate the skeleton sketch’s display. After the MoCap data (BVH file) has been uploaded, the evaluation is performed by computing the RULA scores and showing them in the Score List area. As seen in Figure B.4, various RULA scores may be added and
B. Web-based and Android-based applications for ergonomic evaluation
Figure B.4: User interface of the web-based application. displayed in the scores list area. A score can be selected by either selecting it on the score dropdown list or selecting the joint of the body region that is wished to assess. There is a settings menu for selecting how scores should be computed. For instance, if external forces are present or indicate the movement’s repetition. In the External Factors section of the web-based application, illustrated in Figure B.6a, the default values set for the computing of the Legs, Muscle Use, and Force scores can be modified. First, it must be indicated if the human has any support on the legs and feet while doing the movement under analysis. Then, for the computation of the Muscle use and Force scores, it is necessary to indicate if the work posture is static, repeated in certain periods, or intermittent. If it is a static posture, the module will request the duration the human spends in that posture. If the posture is intermittent or repeated over a period of time, the module will request the number of repetitions that the human performs in one minute. In addition, the load the human is subjected to during the movement can also be specified. The effect of these manually set parameters on the overall RULA score is indicated in [McAtamney, 1993]. These parameters can be similarly modified in the Android application in the section RULA computation, illustrated in Figure B.6b. The skeleton sketch and animation are organized in tabs. For these visualizations, the skeleton posture of each frame was obtained by using the segmented and initial posture offsets provided by the BVH file. The skeleton sketch illustrates the worker’s posture on different frames according to the parameters set in the visual control section. The postures shown on 146 B.2.
Automatic ergonomic evaluation module
Figure B.5: User interface of the android application. (b) (a) Figure B.6: Settings menu for adjusting manual parameters. (a) Web-based application; (b) Android application. ergonomi evaluation
Figure B.7: Skeleton sketch with color-coded scores.
the skeleton sketch match the color-coded scores, which are aligned with the timeline of the recording. The animation displays the video of the uploaded motion data as well as the buttons for pausing and playing it. When the cursor is positioned over a score annotation in the score list, its colors are placed on the background of the skeleton sketch with the end to better visualize the scoring for each selected frame. In addition, as shown in Figure B.7, the most critical joint for this score annotation is emphasized in red. When the cursor is put over the Upper Left Arm annotation, for instance, the left shoulder joint is highlighted because the angle ranges for this score assignment are with respect to this joint. In addition to the score and colors, the angles assessed during the RULA evaluation are also presented on the score annotation (note the angles displayed in the color bars in Figure B.7).
Figure B.7 illustrates the increase in ergonomic risk as the task progresses, as indicated by the Left Final RULA score.
This visualization demonstrates that when the arm is raised, the ergonomic risk of the posture increases. This event is clearly denoted by the transition from the color yellow to red and the increase in angle depicted on its color annotation (Upper Left Arm). B.3 Conclusion and future work
The developed applications are intended to be a useful tool for analysts, facilitating the evaluation of workers’ exposure to ergonomic risk factors related to WMSDs2. The proposed solution for automatic ergonomic evaluation utilizing MoCap data offers a number of advantages and presents interesting perspectives for ergonomists, factory production directors, workers, and anyone else interested in movement analysis. These applications could permit recording and storing analysis results from different executions of the same movement. From the ergonomic perspective, the applications allow comparing results and monitoring workers’ performance to detect any progress or regress. If progress can be observed in this intrapersonal performance study, the analyst may attempt to uncover aspects that bring the worker to 2 Note to reviewers: The applications have not yet been released as they are still under testing. 148
B.3. Conclusion and future work improvement.
In order to enrich the application, its compatibility with motion data recorded with other acquisition systems could be implemented. This by taking into consideration the diverse issues that might be faced (occlusions, noise, inaccurate data, etc.). In addition to the ergonomic evaluation, other human movement analyses, such as the dexterity analysis presented in Chapter 5, could be implemented in the applications. The motion representations could be learned using cloud services connected to the application.
Brenda Elizabeth Olivas-Padilla, Sotiris Manitsaris, Dimitrios Menychtas, and Alina Glushkova. "Stochastic-biomechanic modeling and recognition of human movement primitives, in industry, using wearables". Sensors, 2021. DOI: https://doi.org/10.3390/s21072497 Brenda Elizabeth Olivas-Padilla, Sotiris Manitsaris, and Alina Glushkova. "Motion Capture Benchmark of Real Industrial Tasks and Traditional Crafts for Human Movement Analysis". IEEE Access, 2023. in
Brenda Elizabeth Olivas-Padilla, Dimitris Papanagiotou, Gavriela Senteri, Sotiris Manitsaris, and Alina Glushkova. "Improving Human-Robot Collaboration in TV assembly through computational ergonomics: effective task delegation and robot adaptation". The IEEE International Conference on Systems, Man, and Cybernetics (SMC), Hawaii, USA, 2023 - Under review. Brenda Elizabeth Olivas-Padilla and Sotiris Manitsaris. "Deep state-space modeling for explainable representation, analysis, and generation of professional human movements". Pending. International conferences Agnès Aubert, Brenda Elizabeth Olivas-Padilla, Vasileios Syrris, and Sotiris Manitsaris. "Deep learning architectures applied for recognizing human motion primitives from the Ergonomic Assessment Worksheet". ICRA workshop - Unlocking the potential of human-robot collaboration for industrial applications, Xi’an, China, 2021. Brenda Elizabeth Olivas-Padilla, Dimitrios Menychtas, Alina Glushkova, and Sotiris Manitsaris. "Hidden Markov modelling and recognition of Euler-based motion patterns for automatically detecting risks factors from the European assembly worksheet". The 27th IEEE International Conference on Image Processing (ICIP), Abu Dhabi, United Arab Emirates, 2020. DOI: https://doi.org/10.1109/ICIP40778.2020.9190756 Brenda Elizabeth Olivas-Padilla, Alina Glushkova, Dimitrios Menychtas, and Sotiris Manitsaris. "Designing a web-based Automatic Ergonomic Assessment using Motion Data". Proceedings of the 12th ACM International Conference on PErvasive Technologies Related to Assistive Environments (PETRA), Rhodes, Greece, 2019. DOI: https://doi.org/10.1145/3316782.3322758 Brenda Elizabeth Olivas-Padilla, Alina Glushkova, and Sotiris Manitsaris. "Motion analysis for identification of overused body segments: the packaging task in industry 4.0 ". 17th IFIP TC 13 International Conference of Human-Computer Interaction (INTERACT), Paphos, Cyprus, 2019. Chapitre 1 L’introduction décrit le contexte de cette thèse en donnant un aperçu général de la recherche sur l’analyse du mouvement humain ainsi que de ses défis actuels. Les hypothèses et objectifs formulés sont présentés, ainsi qu’un résumé des contributions et la structure de la thèse. Cette thèse se concentre sur la modélisation analytique de la dynamique du mouvement humain. Elle explore l’estimation des paramètres du mouvement dans le but de développer une méthode généralisée de compréhension du mouvement. L’analyse est effectuée sur des patrons de mouvement globaux (corps entier) plutôt que seulement sur des patrons locaux tels que les gestes de la main ou les expressions faciales. Pour une simulation réaliste du mouvement humain, le modèle de transition approprié a été recherché, qui devrait également décrire le phénomène ou les liens entre les suppositions spatiales et temporelles. Les suppositions spatiales doivent tenir compte des interdépendances potentielles entre les articulations de la structure squelettique articulée. D’autre part, les suppositions temporelles impliquent le principe fondamental que la plupart des séries temporelles, telles que les mouvements humains, présentent intrinsèquement, qui est la dépendance entre les observations adjacentes. Par conséquent, le modèle proposé doit être une représentation suffisamment précise du système dynamique qu’est le mouvement humain pour atteindre les objectifs précédents (simulation précise du mouvement humain et compréhension de la réalisation du mouvement). Les deux hypothèses suivantes ont été formulées pour guider cette recherche : Hypothèse 1 La dynamique du mouvement humain peut être modélisée de manière analytique en tenant compte de la stochastique du mouvement et de la structure physique du corps humain. Hypothèse 2 L’association des articulations du corps
et
leur
contribution
pendant l’exécution d’un mouvement humain peuvent être apprises et représentées par des modèles interprétables.
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AttentionFire_v1.0: interpretable machine learning fire model for burned area predictions over tropics
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Correspondence: Qing Zhu (qzhu@lbl.gov) Received: 26 July 2022 – Discussion started: 11 August 2022 Revised: 20 December 2022 – Accepted: 7 January 2023 – Published: 3 February 2023 Revised: 20 December 2022 – Accepted: 7 January 2023 – Published: 3 February 2023 Abstract. African and South American (ASA) wildfires
account for more than 70 % of global burned areas and
have strong connection to local climate for sub-seasonal
to seasonal wildfire dynamics. However, representation of
the wildfire–climate relationship remains challenging due to
spatiotemporally heterogenous responses of wildfires to cli-
mate variability and human influences. Here, we developed
an interpretable machine learning (ML) fire model (Atten-
tionFire_v1.0) to resolve the complex controls of climate
and human activities on burned areas and to better predict
burned areas over ASA regions. Our ML fire model substan-
tially improved predictability of burned areas for both spatial
and temporal dynamics compared with five commonly used
machine learning models. More importantly, the model re-
vealed strong time-lagged control from climate wetness on
the burned areas. The model also predicted that, under a high-
emission future climate scenario, the recently observed de-
clines in burned area will reverse in South America in the
near future due to climate changes. Our study provides a re-
liable and interpretable fire model and highlights the impor-
tance of lagged wildfire–climate relationships in historical
and future predictions. a Li1,2, Qing Zhu1, William J. Riley1, Lei Zhao3, Li Xu4, Kunxiaojia Yuan1,2, Min Chen5, Hua
anya Gong6, and James T. Randerson4 1Climate and Ecosystem Sciences Division, Climate Sciences Department, Lawrence Berkeley National Laboratory,
Berkeley, CA, USA 1Climate and Ecosystem Sciences Division, Climate Sciences Department, Lawrence Berkeley National Laboratory,
Berkeley, CA, USA 1Climate and Ecosystem Sciences Division, Climate Sciences Department, Lawrence Berkeley National Laboratory,
B k l
CA USA 2State Key Laboratory of Information Engineering in Surveying, Mapping and Remote Sensing,
Wuhan University, Wuhan, China 2State Key Laboratory of Information Engineering in Surveying, Mapping and Remote Sensing,
Wuhan University, Wuhan, China 3Department of Civil and Environmental Engineering, University of Illinois Urbana-Champaign, Champaign, IL, USA
4Department of Earth System Science, University of California Irvine, Irvine, CA, USA 3Department of Civil and Environmental Engineering, University of Illinois Urbana-Champaign, C
4Department of Earth System Science, University of California Irvine, Irvine, CA, USA 3Department of Civil and Environmental Engineering, University of Illinois Urbana-Champaign, Champaign, IL, USA
4Department of Earth System Science, University of California Irvine, Irvine, CA, USA 5Department of Forest and Wildlife Ecology, University of Wisconsin-Madison, Madison, WI, USA
6School of Remote Sensing and Information Engineering, Wuhan University, Wuhan, China 5Department of Forest and Wildlife Ecology, University of Wisconsin-Madison, Madison, WI, USA
6School of Remote Sensing and Information Engineering, Wuhan University, Wuhan, China Department of Forest and Wildlife Ecology, University of Wisconsin Madison, Madison, WI, USA
6School of Remote Sensing and Information Engineering, Wuhan University, Wuhan, China Model description paper Model description paper Geosci. Model Dev., 16, 869–884, 2023
https://doi.org/10.5194/gmd-16-869-2023
© Author(s) 2023. This work is distributed under
the Creative Commons Attribution 4.0 License. Published by Copernicus Publications on behalf of the European Geosciences Union. AttentionFire_v1.0: interpretable machine learning fire model for
burned-area predictions over tropics Fa Li1,2, Qing Zhu1, William J. Riley1, Lei Zhao3, Li Xu4, Kunxiaojia Yuan1,2, Min Chen5, Huayi Wu2, Zhipeng Gui6,
Jianya Gong6, and James T. Randerson4 F. Li et al.: AttentionFire_v1.0 F. Li et al.: AttentionFire_v1.0 fraction, and livestock density, wildfire still plays a signifi-
cant role in mediating surface climate (Xu et al., 2020), bio-
geochemical cycles, and human health (Andela et al., 2017). Further, 21st century projections of increases in temperature,
regional drought (Dai, 2013; Taufik et al., 2017), and precip-
itation variations may outweigh these direct human impacts
and result in unprecedentedly fire-prone environments over a
large fraction of Africa (Van Der Werf et al., 2008; Andela
and Van Der Werf, 2014; Archibald et al., 2009) and South
America (Pechony and Shindell, 2010; Malhi et al., 2008). These factors highlight the need for better understanding,
prediction, and management of these critical fire regions to
minimize economic losses, human health hazards, and natu-
ral ecosystem degradation. Therefore, improved understand-
ing and accurate prediction of wildfire activity is increasingly
important for effective fire management and sustainable de-
cision making. tions and fire management policies may also affect the fire–
climate relationships (Andela et al., 2017). Therefore, strong
climate controls from wet season to dry season need to be
considered along with fuel distributions and human activities
for continental fire predictions under climate change. p
g
Accurate predictive modeling of wildfire with skillful rep-
resentation of how environmental and anthropogenic factors
modulate the burned area is still challenging. State-of-the-art
process-based fire models (e.g., the Fire Model Intercompar-
ison Project; Rabin et al., 2017) have reasonably simulated
the spatial distribution of burned areas. However, they gen-
erally do not accurately capture burned-area seasonal vari-
ation and inter-annual trends and variability (Andela et al.,
2017). Improving predictability and reducing uncertainties of
process-based models require more sophisticated representa-
tion of fire processes and parameterization, which remain a
long-term challenge (Bowman et al., 2009; Hantson et al.,
2016; Teckentrup et al., 2019). In response to this challenge,
data-driven statistical or machine learning (ML) approaches
have been developed and demonstrated to effectively cap-
ture wildfire severity and burned-area dynamics (Archibald
et al., 2009; Chen et al., 2020, 2011; Zhou et al., 2020). However, the spatially heterogenous, non-linear, and time-
lagged controls have been oversimplified, e.g., using linear
models or only considering climate variables at specific time
lags or seasons (Chen et al., 2011, 2016, 2020; Archibald
et al., 2009; Gray et al., 2018) or have been black boxed. F. Li et al.: AttentionFire_v1.0 For example, the commonly used neural network or deep-
learning models (Zhu et al., 2022; Joshi and Sukumar, 2021)
themselves are complex and built upon hidden neural lay-
ers with non-linear activation functions and thus cannot di-
rectly identify the relative importance of different drivers for
wildfires (Murdoch et al., 2019; Jain et al., 2020). A few ML
models (e.g., decision tree and random forest) provide vari-
able importance; however, such importance scores are con-
stant across the entire dataset rather than spatiotemporally
varied (S. S. C. Wang et al., 2021; Yuan et al., 2022b). While
post-hoc analyses could interpret ML models (Altmann et
al., 2010; Lundberg and Lee, 2017), inconsistent and unsta-
ble explanations can be derived with different post-hoc meth-
ods or settings (Slack et al., 2021; Molnar et al., 2020). Such
limitations impede an interpretable and reliable way to un-
derstand the critical spatiotemporal processes from wet sea-
son to dry season (Reichstein et al., 2019; Jain et al., 2020). Climate is acknowledged as one of the most dominant con-
trollers of ASA wildfires (Chen et al., 2011; Andela et al.,
2017). For example, precipitation variations contribute sub-
stantially to burned-area patterns in southern and northern
Africa (Andela and Van Der Werf, 2014; Archibald et al.,
2009) and are also closely linked to wildfire spatiotemporal
dynamics in South America (Chen et al., 2011; Van Der Werf
et al., 2008; Malhi et al., 2008). More importantly, the strong
controls of climate on wildfires often show time lags, and the
time delay can be on the order of multiple months (Van Der
Werf et al., 2008; Andela and Van Der Werf, 2014). Mean-
while, ocean dynamics (e.g., El Niño–Southern Oscillation,
ENSO) may also exert considerable influences on ASA wild-
fires through influencing wet- and wet-to-dry-season climate
and fuel conditions (Yu et al., 2020; Chen et al., 2016; An-
dela and Van Der Werf, 2014; Chen et al., 2011, 2017). The
time lags between ocean dynamics and wildfires can be even
longer than that between climate and wildfires (Chen et al.,
2020), which enables wildfire predictions ahead of fire sea-
son (Chen et al., 2011, 2016, 2020; Turco et al., 2018). The
spatiotemporal responses of wildfires to climate changes are
complicated by non-linear interactions among climate, veg-
etation, and human activities (Van Der Werf et al., 2008;
Andela et al., 2017). 1
Introduction Wildfires modify land surface characteristics, such as veg-
etation composition, soil carbon, surface runoff, and albedo,
with significant consequences for regional carbon, water, and
energy cycles (Benavides-Solorio and MacDonald, 2001;
Shvetsov et al., 2019; Randerson et al., 2006). Over African
and South American (ASA) regions, where more than 70 %
of global burned area occurs, wildfires emit ∼1.4 PgC yr−1
(∼65 % of global wildfire emissions; van Der Werf et al.,
2017) and dust and aerosols that can alter regional cli-
mate through radiative processes (Etminan et al., 2016; Ra-
manathan et al., 2001; van Der Werf et al., 2017). While
greenhouse gas emissions contribute to climate change, other
toxic species and airborne particulate matter from wildfires
lead to substantial health hazards, including elevated prema-
ture mortality (Knorr et al., 2017; Lelieveld et al., 2015). In
particular, wildfire particulate matter emissions across tropi-
cal regions have exceeded current anthropogenic sources and
are predicted to dominate future regional emissions (Knorr et
al., 2017). Although total tropical wildfire-burned area has declined
over the past few decades due to climate change and human
activities (Andela and Van Der Werf, 2014; Andela et al.,
2017), e.g., from increases in population density, cropland Published by Copernicus Publications on behalf of the European Geosciences Union. 870 2.1
AttentionFire model The weight or importance of the ith driving vari-
able wi is calculated as wi′ = tanh(Wa[hi
sum,hi
T ])
wi =
ewi ′
Pn
j=1ewj ′ . (4) (4) Finally, using the weighted sum of all driving variables, the
model generates the prediction ˆYT +1 : Finally, using the weighted sum of all driving variables, the
model generates the prediction ˆYT +1 : oi = Wo[hi
sum,hi
T ] + bo
ˆYT +1 =
n
X
i=1
oiwi,
(5) oi = Wo[hi
sum,hi
T ] + bo oi = Wo[hi
sum,hi
T ] + bo oi = Wo[hi
sum,hi
T ] + bo Given
four
categories
of
time
series,
X =
(Xl,Xs,Xf,Xc)T , where T is the length of time series, we
use Xi = (xi
1,xi
2,..., xi
T )T ∈RT , where 1 ≤i ≤n, to denote
the ith time series, and we use Xt = (x1
t ,x2
t ,..., xn
t )T ∈Rn,
where 1 ≤t ≤T , to represent the vector at time step t. xI
t ,
xs
t , xf
t , and xc
t represent the variables of ignition (e.g., pop-
ulation density), suppression (e.g., road network density),
fuel availability (e.g., living biomass), and climate (e.g.,
precipitation) at time step t. The AttentionFire model aims
to learn a nonlinear function F to map the n time series to
the observed burned area YT +1 at time step T + 1: i
o[
sum
T ]
o
ˆYT +1 =
n
X
i=1
oiwi,
(5) (5) where Wa ∈R1×2m is a learnable parameter matrix, and the
linear function with weight Wo ∈Rm and bias bo ∈R, along
with attention-calculated weight wi, produce the final pre-
diction result. The parameters of attention-based LSTM are
learned via a back-propagation algorithm by minimizing the
mean-squared error between predictions and observations
(Guo et al., 2019; Leung and Haykin, 1991). where Wa ∈R1×2m is a learnable parameter matrix, and the
linear function with weight Wo ∈Rm and bias bo ∈R, along
with attention-calculated weight wi, produce the final pre-
diction result. The parameters of attention-based LSTM are
learned via a back-propagation algorithm by minimizing the
mean-squared error between predictions and observations
(Guo et al., 2019; Leung and Haykin, 1991). ˆYT +1 = F(Xl,Xs,Xf,Xc)T ,
(1) (1) The AttentionFire model is implemented with Python un-
der Python 3 environment. 2.1
AttentionFire model (2) The AttentionFire model is based on an interpretable
attention-augmented LSTM (Liang et al., 2018; Qin et al.,
2017; Guo et al., 2019; Li et al., 2020; Vaswani et al.,
2017) framework. Like the traditional artificial neural net-
work (ANN) models, the LSTM is also built upon neu-
rons and the non-linear activation functions; specifically, the
LSTM uses the gating mechanism (i.e., forget, input, and out-
put gates) (Hochreiter and Schmidhuber, 1997; Wang and
Yuan, 2019) to filter out useless information while keeping
useful information underlying in the time series as hidden
states (Fig. 1). Relative to traditional ANN, the LSTM has
shown advantages in capturing short- and long-term depen-
dencies in input time series (Hochreiter and Schmidhuber,
1997), such as the time-lagged controls from wet-to-dry-
season climate conditions on wildfires. However, LSTM can-
not explicitly and dynamically select important drivers from
multiple driving time series to make predictions (Qin et al.,
2017; Liang et al., 2018; Guo et al., 2019; Li et al., 2020;
Vaswani et al., 2017). Further, LSTM works as a black box,
lacking interpretability to identify the relative importance of
each driver across different time steps (Guo et al., 2019; Li
et al., 2020; Liang et al., 2018). Attention mechanisms over-
come these challenges by adaptively assigning larger weights
to more important drivers and time steps (Liang et al., 2018;
Vaswani et al., 2017). Here, we use attention mechanisms to
explicitly capture controlling factors of fire predictions with
various time lags (Fig. 1). Below are detailed descriptions of
the fire model. where hi
t ∈Rm is the hidden state vector of the ith driving
series at time step t that stores the summary of the past in-
put sequence (Hochreiter and Schmidhuber, 1997); wi
t is the
calculated weight for the ith driver at time step t through at-
tention function fattn: wi
t
′ = tanh(Wphi
t)
wi
t =
ewi
t ′
PT
j=1ewj
t
′ ,
(3) (3) where Wp ∈R1×m is a parameter matrix that needs to be
learned. To further capture the relative importance of the ith driving
variable compared to other driving variables, variable atten-
tion is used for the summarized information hi
sum and hi
T . Note that hi
T is also a kind of summarized information de-
rived by the LSTM (Hochreiter and Schmidhuber, 1997; Guo
et al., 2019). 2
Methods wi
t = fattn(hi
t)
hi
sum =
T
X
t=1
wi
thi
t,
(2) wi
t = fattn(hi
t)
hi
sum =
T
X
t=1
wi
thi
t, F. Li et al.: AttentionFire_v1.0 871 Emission Database (GFED) and compared it with five other
machine-learning-based fire models. tention is applied to hi
t to calculate its corresponding weight
or importance wi
t. Third, the weighted summation hi
sum of hi
t
is obtained to represent the summarized information for the
ith driving variable: F. Li et al.: AttentionFire_v1.0 In more xeric subtropical regions, in-
creasing precipitation during the wet season can be the dom-
inant controller on increasing wildfire during the following
dry season (through regulation of fuel availability and fuel
spatial structures) (Van Der Werf et al., 2008; Littell et al.,
2009; Archibald et al., 2009). In contrast, increasing precip-
itation in more mesic regions results in excessive fuel mois-
ture, thereby becoming the main limitation of dry-season
wildfires (i.e., opposite fire trends are observed with increas-
ing precipitation in northern and southern Africa) (Van Der
Werf et al., 2008; Andela and Van Der Werf, 2014). In addi-
tion to natural processes, human activities are primary igni-
tion sources and have shaped fire patterns in the ASA regions
(Aragao et al., 2008; Archibald et al., 2009; Andela et al.,
2017). Fire-use types driven by local socio-economic condi- In this work, we developed a wildfire model (Attention-
Fire) leveraging on an interpretable long short-term mem-
ory (LSTM) framework to predict wildfire burned areas over
northern hemispheric Africa (NHAF), southern hemispheric
Africa (SHAF), and southern hemispheric South America
(SHSA) (Giglio et al., 2013). We also focused on using
the AttentionFire model to explore the dependency of simu-
lated burned area on different drivers from wet season to dry
season across different grid cells. We assessed model pre-
dictability with observed burned area from the Global Fire https://doi.org/10.5194/gmd-16-869-2023 Geosci. Model Dev., 16, 869–884, 2023 https://doi.org/10.5194/gmd-16-869-2023 2.2
Baseline models and model settings impede detailed interpretation of the variable importance,
like over space and time. The aforementioned ML models
have been commonly used in wildfire science (Jain et al.,
2020). Five other widely used machine learning (ML) models are
used as baseline models to compare with AttentionFire
model: ANN (Joshi and Sukumar, 2021; Zhu et al., 2022),
decision tree (DT) (Amatulli et al., 2006; Coffield et al.,
2019), random forest (RF) (Yu et al., 2020; Li et al., 2018;
Gray et al., 2018), gradient-boosting decision tree (GBDT)
(Coffield et al., 2019; Jain et al., 2020), and naive LSTM
(Liang et al., 2019; Natekar et al., 2021; Gui et al., 2021;
Mei and Li, 2019). The details of the baseline models se-
lected, including strengths, potential limitations, and their
applications in wildfire studies, and references are listed in
Table 1. The ANN and LSTM have shown good performance
on multiple earth science problems (Yuan et al., 2022a; Re-
ichstein et al., 2019), including wildfires (Joshi and Suku-
mar, 2021; Liang et al., 2019; Zhu et al., 2022); however,
the black-box nature of such models makes them lack inter-
pretability. The DT method provides variable importance and
is easily interpretable with its single-tree structure, but it is
prone to overfitting compared to RF and GBDT. The RF alle-
viates the overfitting through feature selection and ensemble
learning (Breiman, 2001), while the GBDT avoids overfit-
ting by constructing multiple trees with shallow depth (Ke et
al., 2017). DT, RF, and GBDT provide variable importance
scores for dominant driver inference; however, such impor-
tance scores are constant across the entire dataset and thus )
The inputs of climate- and fuel-related variables for the
first four models (non-sequence models) are variables of the
latest three months available for prediction (Yu et al., 2020),
while the corresponding inputs of naive LSTM and Atten-
tionFire models are whole-year historical time sequences
which cover dynamics from wet to dry seasons to capture
short- and long-term dependencies underlying the input se-
quence (Qin et al., 2017; Vaswani et al., 2017; Guo et al.,
2019; Li et al., 2020). The socioeconomic predictors (i.e.,
population, road density, livestock) consider only the more
recent and available statistics typically reported at a year
scale. 2.1
AttentionFire model The model is open access at https:
//doi.org/10.5281/zenodo.7416437 (Li et al., 2022b) under
Creative Commons Attribution 4.0 International license. De-
tailed code and descriptions are included in the repository,
including loading datasets, model initialization, training, pre-
dicting, saving parameters, and loading the trained model
(see more details in “Code availability” section). where ˆYT +1 is the predicted burned area at time step T + 1. where YT +1 is the predicted burned area at time step T + 1. First, the model iteratively transforms the ith driving vari-
able at time step t to a hidden state vector hi
t, where 1 ≤t ≤
T and 1 ≤i ≤n, through LSTM gate mechanisms (please re-
fer to Li et al., 2020, for the details of the gates in Fig. 1). Sec-
ond, as the importance of each time step varies, temporal at- https://doi.org/10.5194/gmd-16-869-2023 Geosci. Model Dev., 16, 869–884, 2023 872 F. Li et al.: AttentionFire_v1.0 _
Figure 1. An illustrative workflow for AttentionFire_v1.0 model prediction. Four kinds of drivers are considered: ignition related, suppression
related, fuel, and climate. The temporal attention is used to identify important time steps for each kind of driver, while the variable attention
is used to identify important drivers for final burned-area prediction. Figure 1. An illustrative workflow for AttentionFire_v1.0 model prediction. Four kinds of drivers are considered: ignition related, suppression
related, fuel, and climate. The temporal attention is used to identify important time steps for each kind of driver, while the variable attention
is used to identify important drivers for final burned-area prediction. 2.2
Baseline models and model settings For each model, we iteratively leave a 1-year dataset
(one out of all 19 years’ datasets for the period 1997–2015,
∼5 % of all datasets) out (i.e., a holdout dataset, such as the
dataset in 2015, that the model has never seen) for testing,
1 year of data (∼5 % of all datasets, such as the dataset in
2014) for validation (the model was stopped for training, and
its parameters were saved when it showed the highest per-
formance on the validation dataset to avoid overfitting dur-
ing training; Yuan et al., 2022b; Jabbar and Khan, 2015) and
use the remaining dataset (∼90 % of all datasets, such as
the dataset during 1997–2013) for model training (i.e., tun-
ing model parameters). Such an evaluation scheme quanti- https://doi.org/10.5194/gmd-16-869-2023 Geosci. Model Dev., 16, 869–884, 2023 873 F. Li et al.: AttentionFire_v1.0 Table 1. Strengths, potential limitations, and applications of selected baseline models in wildfire studies. Model (acronym)
Strengths
Potential limitations
Applications
Random forest (RF)
(Breiman, 2001)
Provides variable importance;
alleviates overfitting through feature se-
lection and ensemble learning
Constant variable importance rather than var-
ied; time-consuming when building large trees;
may not perform well on time series with lags
Gray et al. (2018);
Yu et al. (2020)
Decision tree (DT)
(Safavian and
Landgrebe, 1991)
Provides variable importance; easily in-
terpretable with its single-tree structure
Prone to overfitting; constant variable impor-
tance rather than varied; time-consuming when
building a large tree; may not perform well on
time series with lags
Amatulli et al. (2006);
Coffield et al. (2019)
Gradient-boosting
decision tree (GBDT)
(Ke et al., 2017)
Alleviates overfitting by building multi-
ple shallow trees; generally fast because
of the shallowness of each tree built
Constant variable importance rather than var-
ied; may not perform well on time series with
lags
Coffield et al. (2019);
Jain et al. (2020)
Artificial neural
network (ANN)
(Ke et al., 2017)
Shows good performance on com-
plex and non-linear problems; allevi-
ates overfitting through techniques like
dropout and regularization
Lack of interpretability; hard to know the op-
timal neural network structures for different
problems
Joshi and Sukumar (2021);
Zhu et al. 2.2
Baseline models and model settings (2022)
Long short-term mem-
ory (LSTM)
(Hochreiter and
Schmidhuber, 1997)
Shows good performance on time se-
ries predictions; alleviates overfitting
through techniques like dropout and
regularization
Lack of interpretability; may not be suitable
for non-time-series problems; vanishing gradi-
ent problem when deployed to long time series
(Li et al., 2020; Liang et al., 2018)
Liang et al. (2019);
Natekar et al. (2021) Table 1. Strengths, potential limitations, and applications of selected baseline models in wildfire studies. teractions and thus the wet-, wet-to-dry-, and onset-of-dry-
season climate in South America (Chen et al., 2011). The two
indices were significantly correlated with peak fire month
wildfires 3 to 7 months later and could predict fire season
wildfires in many regions of South America with lead times
of 3 to 5 months (Chen et al., 2011). The controls of SST
anomalies in the tropical Pacific on climate and thus on wild-
fires were also found in northern and southern Africa (An-
dela and Van Der Werf, 2014). In addition, SST anomalies in
the tropical northern and southern Atlantic could also affect
wildfires in South America (Chen et al., 2016) and Africa (Yu
et al., 2020; Chen et al., 2020). Therefore, we included ocean
indices (Table 2) and investigated their impacts on wildfire
predictions with the AttentionFire model (see Sect. 3.4). fied model performance on deducing the temporal dynamics
of fires at the annual scale, which is critical for future projec-
tions, while leveraging as much data as possible for model
training. Details of the settings for used models in the exper-
iments are listed in Table S1. Geosci. Model Dev., 16, 869–884, 2023 2.3
Datasets and experiments The satellite-based global burned-area dataset (Global Fire
Emissions Database; Giglio et al., 2013) is used as prediction
target, and datasets of various socio-environmental drivers
are used as model inputs. Population density, livestock den-
sity, road-network density, and land use are considered as an-
thropogenic factors on fire ignition and spread. Fuel variables
include fuel moisture and live- and dead-vegetation biomass. Seven meteorology variables from National Centers for Envi-
ronmental Prediction–Department of Energy (NCEP–DOE)
Reanalysis are considered, including air temperature, pre-
cipitation, surface pressure, wind speed, specific humidity,
downward shortwave radiation, and vapor pressure deficit. Details of each dataset and corresponding references are
listed in Table 2. The raw datasets were unified to the same
spatial resolution (T62 resolution: ∼210 km at the Equator)
at the monthly scale, with a covering period from 1997 to
2015. For future projection (2016–2055) of burned area with
the AttentionFire model, land use changes (Hurtt et al.,
2020), population growth, projected climate, and fuel from
five fully coupled Earth system model (ESM) simulations of
CMIP6 (O’Neill et al., 2016) under low- (SSP126) and high-
emission (SSP585) scenarios were used as the ML model
input, respectively. The reason to select 2016–2055 as the
projected period was that, during 2016–2055, the 99th per-
centiles of precipitation, temperature, and vapor pressure
deficit were within the range of corresponding historical ob-
servations, which means that the trained model has covered
the range of most projected drivers in the near future and can
alleviate extrapolation uncertainty caused by climate change. We also made a longer projection till the end of 21st century
and analyzed its longer-term trend (see Sect. 3.4). All avail-
able ESMs with outputs of historical and future (SSP126 and
SSP585) fuel availability (i.e., biomass of coarse wood de-
bris, vegetation, and litter) and climate variables (Table 2) In addition to the local socio-environmental drivers, we
also explored the impacts of ocean indices on burned-area
predictions. Chen et al. (2011) found that wildfires in South
America were closely linked to the Oceanic Niño Index
(ONI) and the Atlantic Multidecadal Oscillation (AMO) in-
dex. The ONI and AMO reflected the sea surface temperature
(SST) anomalies in the tropical Pacific and north Atlantic. The SST anomalies directly affected ocean–atmosphere in- https://doi.org/10.5194/gmd-16-869-2023 Geosci. Model Dev., 16, 869–884, 2023 F. Li et al.: AttentionFire_v1.0 874 Table 2. Input and output variables and datasets of the AttentionFire model. Table 2. 3
Results and discussions were selected, including ACCESS-ESM1-5 (Ziehn et al.,
2020), CESM2 (Danabasoglu et al., 2020), NorESM2-LM
(Seland et al., 2020), NorESM2-MM (Seland et al., 2020),
and TaiESM1 (Y. C. Wang et al., 2021). For each ESM, the
variable bias was corrected with the mostly used linear scal-
ing method (Maraun, 2016; Dangol et al., 2022; Shrestha
et al., 2017), which adjusted the bias in model simulations
based on the ratio of modeled- and observed-variable mean
value. Then the bias-corrected variables of each ESM were
used to drive the AttentionFire model for future burned-
area projection. Finally, given the uncertainty of each ESM,
the multi-model ensemble (MME) mean of projected burned
area was calculated (Li et al., 2022a) and analyzed. Details of
the bias correction method can be found in Maraun (2016). For future projections, temporally constant road and live-
stock density were used due to the lack of future data in the
two scenarios (i.e., SSP585 and SSP126), and the Attention-
Fire model was not coupled in the ESMs. Such limitations
and uncertainties were discussed in Sect. 3.5. were selected, including ACCESS-ESM1-5 (Ziehn et al.,
2020), CESM2 (Danabasoglu et al., 2020), NorESM2-LM
(Seland et al., 2020), NorESM2-MM (Seland et al., 2020),
and TaiESM1 (Y. C. Wang et al., 2021). For each ESM, the
variable bias was corrected with the mostly used linear scal-
ing method (Maraun, 2016; Dangol et al., 2022; Shrestha
et al., 2017), which adjusted the bias in model simulations
based on the ratio of modeled- and observed-variable mean
value. Then the bias-corrected variables of each ESM were
used to drive the AttentionFire model for future burned-
area projection. Finally, given the uncertainty of each ESM,
the multi-model ensemble (MME) mean of projected burned
area was calculated (Li et al., 2022a) and analyzed. Details of
the bias correction method can be found in Maraun (2016). For future projections, temporally constant road and live-
stock density were used due to the lack of future data in the
two scenarios (i.e., SSP585 and SSP126), and the Attention-
Fire model was not coupled in the ESMs. Such limitations
and uncertainties were discussed in Sect. 3.5. 2.3
Datasets and experiments Input and output variables and datasets of the AttentionFire model. Variable category
Variables (abbreviation, units)
Spatial (temporal) resolution
Dataset and reference
Wildfire
Burned area (BA, ha per month)
0.25◦(monthly)
Global Fire Emissions
Database 4
(Giglio et al., 2013)
Climate
Precipitation (RAIN, mm s−1), temperature
(TA, K), surface air pressure (PA, Pa), specific
humidity (SH, kg kg−1), downward shortwave
radiation (SW, W m−2), wind speed (WIND,
m s−1), vapor pressure deficit (VPD, hPa)
(VPD calculated according to Bolton; 1980)
∼1.9◦(monthly)
NCEP–DOE Reanalysis 2
(Kanamitsu et al., 2002)
Fuel conditions
Fuel moisture (FUELM, %), coarse wood de-
bris (CWDC, gC m−2 s−1), vegetation biomass
(VegC, gC m−2 s−1), litter biomass (LitterC,
gC m−2 s−1)
∼1.9◦(monthly)
ELM prognostic simulations
(Zhu et al., 2019)
Human activities
Population density (Popu, persons
per grid)
∼1 km (yearly)
Dobson et al. (2000)
Road density (Road, km km−2)
0.5◦(yearly)
Meijer et al. (2018)
Livestock density (LS, number of livestock per
grid)
0.5◦(yearly)
Rothman-Ostrow et al. (2020)
Land cover
Bare soil (Bare, %), Forest (Forest, %), and
Grass (Grass, %)
0.25◦(yearly)
LUH2 (Hurtt et al., 2020)
Oceanic indices
Ocean Niño Index (ONI), Atlantic Multidecadal
Oscillation (AMO) index, tropical Northern At-
lantic (TNA) index, and tropical Southern At-
lantic (TSA) index
monthly
NOAA climate indices
(Enfield et al., 1999, 2021) Geosci. Model Dev., 16, 869–884, 2023 3.1
Model predictability on burned-area
spatial-temporal dynamics The AttentionFire model accurately captured the spatial dis-
tribution and temporal variations (Figs. 2 and S1) of wildfire-
burned areas over NHAF, SHAF, and SHSA regions. The
AttentionFire model had the lowest mean absolute errors
(MAEs) between model-predicted and observed (GFED)
gridded monthly burned areas among the six ML approaches. The gridded MAEs of burned area for AttentionFire were
110, 142, and 39 Kha yr−1 in NHAF, SHAF, and SHSA re-
gions, which were respectively 6 %–66 %, 13 %–65 %, and
11 %–42% lower than the other five ML approaches in the
three regions. These results highlight the capability of the At-
tentionFire model to capture critical driving factors of burned
area across time and space. The fact that the AttentionFire model outperformed the
other five models (Fig. 2g–i) indicates the benefit of skillfully
integrating time-lagged and spatially heterogenous controls
from critical drivers on wildfires. Compared to non-sequence
models (i.e., RF, MLP, DT, and GBDT), the AttentionFire
model adaptively captured historical dependencies of wild- Geosci. Model Dev., 16, 869–884, 2023 https://doi.org/10.5194/gmd-16-869-2023 875 F. Li et al.: AttentionFire_v1.0 Figure 2. The AttentionFire model accurately captured burned-area spatial dynamics. Spatial distribution of observed and AttentionFire-
predicted fire season mean burned area (BA) with one-month lead time in northern hemispheric African (NHAF) (a–b), southern hemispheric
African (SHAF) (c–d), and southern hemispheric South American (SHSA) (e–f) regions. (g–i) Performance (in terms of mean absolute error
between predicted and observed burned area) of AttentionFire and other five baseline models, including long short-term memory (LSTM),
random forest (RF), artificial neural network (ANN), decision tree (DT), and gradient-boosting decision tree (GBDT). Figure 2. The AttentionFire model accurately captured burned-area spatial dynamics. Spatial distribution of observed and AttentionFire-
predicted fire season mean burned area (BA) with one-month lead time in northern hemispheric African (NHAF) (a–b), southern hemispheric
African (SHAF) (c–d), and southern hemispheric South American (SHSA) (e–f) regions. (g–i) Performance (in terms of mean absolute error
between predicted and observed burned area) of AttentionFire and other five baseline models, including long short-term memory (LSTM),
random forest (RF), artificial neural network (ANN), decision tree (DT), and gradient-boosting decision tree (GBDT). al., 2019; Li et al., 2020) that maximize model predictabil-
ity. Therefore, the variable weights could inform dominant
physical processes, while the temporal weights reflect the
temporal dependency structure, making it interpretable for
spatial-temporal analysis. 3.1
Model predictability on burned-area
spatial-temporal dynamics For the AttentionFire model pre-
dictions, the variable weights showed that climate wetness
exerted strong and spatially heterogenous controls on burned
areas. Specifically, precipitation (for SHAF and SHSA re-
gions) and vapor pressure deficit (VPD; for NHAF region)
played the most important roles (Fig. 3) in burned-area pre-
diction during fire seasons (defined as the four months with
the largest burned areas, Fig. S2), and the control strengths
from those climate wetness variables on fires were signifi-
cantly (one-tailed t test, p value <0.05) stronger in regions
with larger burned areas (grid cells with top 10 % burned ar-
eas) than those with smaller burned areas (grid cells with last
90 % burned areas; Fig. 4a–f). fires on climate conditions from wet to dry seasons (Van Der
Werf et al., 2008; Archibald et al., 2009; Andela and Van
Der Werf, 2014; Chen et al., 2011). A more detailed analy-
sis is provided in next section. Compared to the naive LSTM
models, the variable and temporal attention mechanisms in-
tegrated in AttentionFire has proven to be beneficial to model
performance. The spatial heterogeneity and temporal variation of wild-
fire responses to complex environmental and human fac-
tors have made wildfire predictions challenging, especially
at large spatial scales (Chen et al., 2016; Littell et al., 2016;
Andela and Van Der Werf, 2014; Chen et al., 2011; Zhou et
al., 2020). The capability of the AttentionFire model to rea-
sonably predict spatial and temporal distributions of burned
area ahead of fire season allows more time to explore and im-
plement management options, such as allocation of firefight-
ing resources, fuel clearing, or targeted burning restrictions
(Chen et al., 2011). In AttentionFire model predictions, the precipitation and
VPD explained ∼66 % to ∼80% (Fig. S3) of the annual
mean fire season wildfire-burned areas. Variations of VPD
and precipitation not only affect fire season ignition likeli-
hood and fire spread (Sedano and Randerson, 2014; Holden
et al., 2018) through fuel moisture but also regulate vegeta-
tion growth, fuel structure (e.g., fuel composition and spatial 3.2
Dominant drivers of tropical burned-area
dynamics The AttentionFire model dynamically weights variable im-
portance and highlights critical temporal windows (Qin et
al., 2017; Vaswani et al., 2017; Liang et al., 2018; Guo et https://doi.org/10.5194/gmd-16-869-2023 https://doi.org/10.5194/gmd-16-869-2023 Geosci. Model Dev., 16, 869–884, 2023 876 F. Li et al.: AttentionFire_v1.0 F. Li et al.: AttentionFire_v1.0 Figure 3. Ranked top-five important variables for fire season burned area in northern hemispheric Africa (NHAF) (a), southern hemispheric
Africa (SHAF) (b), and southern hemispheric South America (SHSA) (c). For each grid cell within each study region, there is a mean variable
weight, representing the importance of the variable for fire prediction in the grid cell. For each region, the variable weights are summed,
weighted by their corresponding mean burned areas, and normalized. Figure 3. Ranked top-five important variables for fire season burned area in northern hemispheric Africa (NHAF) (a), southern hemispheric
Africa (SHAF) (b), and southern hemispheric South America (SHSA) (c). For each grid cell within each study region, there is a mean variable
weight, representing the importance of the variable for fire prediction in the grid cell. For each region, the variable weights are summed,
weighted by their corresponding mean burned areas, and normalized. Figure 4. Spatial-temporal importance of climate wetness variables for burned-area dynamics. (a–c) Spatial importance of climate wetness
variables for fire season burned areas. (d–f) Statistical comparison of the climate wetness variable importance over regions with large and
small burned areas. (g–i) Fire season burned-area dependency on the history of the climate wetness driver over northern hemispheric African
(NHAF), southern hemispheric African (SHAF), and southern hemispheric South American (SHSA) regions. Figure 4. Spatial-temporal importance of climate wetness variables for burned-area dynamics. (a–c) Spatial importance of climate wetness
variables for fire season burned areas. (d–f) Statistical comparison of the climate wetness variable importance over regions with large and
small burned areas. (g–i) Fire season burned-area dependency on the history of the climate wetness driver over northern hemispheric African
(NHAF), southern hemispheric African (SHAF), and southern hemispheric South American (SHSA) regions. connectivity; Gale et al., 2021), and fuel availability (Mueller
et al., 2020; Littell et al., 2009, 2016; Van Der Werf et al.,
2008). The importance of these climate wetness variables
confirms the dominant roles of local water balances and air
dryness for wildfire prediction from sub-seasonal to seasonal
scales (Littell et al., 2016; Archibald et al., 2009; Chen et al.,
2011), especially in regions with large burned areas. specific, and these relationships may be useful for develop-
ing, tuning, and benchmarking wildfire models (Zhu et al.,
2022). https://doi.org/10.5194/gmd-16-869-2023 3.3
Possible usage of oceanic index for
long-leading-time predictions ,
)
(
,
)
g
Considering land use changes, population growth, and
projected climate and fuel conditions under the SSP585 high-
emission scenario, our model predicted that burned areas in
the NHAF region will continue to decline; the currently in-
creasing trend will be dampened in the SHAF region, and
the currently decreasing trend will be reversed in the SHSA
region (Fig. 6). The increasing trend in SHSA, decreasing
trend in NHAF, and dampened trend in SHAF under SSP585
were robust when projecting burned area till the end of the
21st century (Fig. S5). Over NHAF and SHSA, burned-area
trends at the grid cell level were mostly robust (Fig. 6a, c;
p<0.05) and of the same sign, thus resulting in a robust
trend at the regional scale. Under the low-emission scenario
(i.e., SSP126), the decreasing trend in NHAF disappeared
(Fig. S5a), and the increasing trend in SHSA was reduced
by ∼69 % (Fig. S5c), implying the big influences of climate
changes and socioeconomic development pathways on future
burn-area changes in the two regions. In ASA regions, large-scale variations of oceanic dynamics
can directly influence local climate (e.g., precipitation varia-
tions during wet seasons; Chen et al., 2011; Andela and Van
Der Werf, 2014) through time-lagged controls of teleconnec-
tions and indirectly influence fires during the following dry
seasons (Chen et al., 2016, 2011; Andela et al., 2017). There-
fore, we hypothesized that ocean dynamics might benefit At-
tentionFire model predictions, especially for long-leading-
time fire predictions, through providing additional informa-
tion that has not been reflected in local climate and land sur-
face conditions (Chen et al., 2016, 2011; Andela et al., 2017;
Chen et al., 2020). We compared model performance for short-term (1–4
months ahead) and long-term (5–8 months ahead) fire pre-
dictions with and without considering the four oceanic in-
dices (OIs). Relative to the MAE of short-term predictions,
the mean MAE of long-term predictions without and with
teleconnections increased by ∼34 % and ∼14 % in NHAF,
∼34 % and ∼15 % in SHAF, and ∼17 % and ∼7 % in
SHSA, respectively, indicating the decline of system pre-
dictability with longer leading time (Fig. 5). However, for
long-term predictions, including OIs could decrease the
mean MAE by ∼20 %, ∼19 %, and ∼11 % in NHAF,
SHAF, and SHSA regions, respectively, compared with the
case without oceanic indices. 3.4
Future trends of burned area over Africa and
South America Due to climate change and human activities (Andela et al.,
2017), strong but opposing trends of burned areas have been
observed in northern (decreasing) and southern (increasing)
hemispheric Africa (Andela and Van Der Werf, 2014) and
within different regions of southern hemispheric America
(Andela et al., 2017) during the recent two decades, resulting
in an overall declining burned-area trend in Africa and South
America. However, whether this decline will persist is under
debate. On the one hand, the projected increases in popula-
tion, expansion of agriculture, mechanized (fire-free) man-
agement, and fire suppression policies will likely continue to
decrease burned areas (Andela and Van Der Werf, 2014), e.g.,
human activities were regarded as one of the main drivers for
fire decline in NHAF region. On the other hand, future cli-
mate change (Dai, 2013; Taufik et al., 2017) could outweigh
human impacts and result in unprecedented fire-prone envi-
ronments in the tropics (Pechony and Shindell, 2010; Malhi
et al., 2008), e.g., fires showed strong dependency on climate
wetness in NHAF, SHAF (Andela and Van Der Werf, 2014;
Archibald et al., 2009) and SHSA (Chen et al., 2011) regions. Considering land use changes, population growth, and
projected climate and fuel conditions under the SSP585 high-
emission scenario, our model predicted that burned areas in
the NHAF region will continue to decline; the currently in-
creasing trend will be dampened in the SHAF region, and
the currently decreasing trend will be reversed in the SHSA
region (Fig. 6). The increasing trend in SHSA, decreasing
trend in NHAF, and dampened trend in SHAF under SSP585
were robust when projecting burned area till the end of the
21st century (Fig. S5). Over NHAF and SHSA, burned-area
trends at the grid cell level were mostly robust (Fig. 6a, c;
p<0.05) and of the same sign, thus resulting in a robust
trend at the regional scale. Under the low-emission scenario
(i.e., SSP126), the decreasing trend in NHAF disappeared
(Fig. S5a), and the increasing trend in SHSA was reduced
by ∼69 % (Fig. S5c), implying the big influences of climate
changes and socioeconomic development pathways on future
burn-area changes in the two regions. 3.4
Future trends of burned area over Africa and
South America i
i
h
d i
f
b
d
h Due to climate change and human activities (Andela et al.,
2017), strong but opposing trends of burned areas have been
observed in northern (decreasing) and southern (increasing)
hemispheric Africa (Andela and Van Der Werf, 2014) and
within different regions of southern hemispheric America
(Andela et al., 2017) during the recent two decades, resulting
in an overall declining burned-area trend in Africa and South
America. However, whether this decline will persist is under
debate. On the one hand, the projected increases in popula-
tion, expansion of agriculture, mechanized (fire-free) man-
agement, and fire suppression policies will likely continue to
decrease burned areas (Andela and Van Der Werf, 2014), e.g.,
human activities were regarded as one of the main drivers for
fire decline in NHAF region. On the other hand, future cli-
mate change (Dai, 2013; Taufik et al., 2017) could outweigh
human impacts and result in unprecedented fire-prone envi-
ronments in the tropics (Pechony and Shindell, 2010; Malhi
et al., 2008), e.g., fires showed strong dependency on climate
wetness in NHAF, SHAF (Andela and Van Der Werf, 2014;
Archibald et al., 2009) and SHSA (Chen et al., 2011) regions. F. Li et al.: AttentionFire_v1.0 F. Li et al.: AttentionFire_v1.0 877 mate (Fig. S4) across the three regions, the OIs did provide
additional information for long-term predictions with lower
biases (Fig. 5). The results demonstrated the potential usage
of teleconnections for long-leading-time burned-area predic-
tions (Chen et al., 2020, 2016, 2011). long time-lagged controls on dry-season burned areas (Abat-
zoglou and Kolden, 2013; Littell et al., 2016; Chen et al.,
2011; Van Der Werf et al., 2008; Archibald et al., 2009; An-
dela and Van Der Werf, 2014). Previous work has shown that when and where fires oc-
curred during the dry season can be affected by precipitation-
induced fuel availability patterns during the wet season and
during wet-to-dry transition seasons in savannah ecosystems
(Van Der Werf et al., 2008; Archibald et al., 2009; Andela
and Van Der Werf, 2014). Also, precipitation variations dur-
ing the wet season and wet-to-dry transition seasons in the
tropical forest ecosystem can affect soil recharge during the
wet season and further affect plant transpiration, local sur-
face humidity, and precipitation during the following dry sea-
son (Chen et al., 2011; Ramos da Silva et al., 2008; Malhi
et al., 2008). The exact responses of fires to short- and long-
term climate variations depend on both local wetness and fuel
conditions (e.g., fires in wetter ecosystems with enough fuel
availability can be mainly limited by the length of dry season,
while fires in drier ecosystems can be limited by fuel avail-
ability during wet season; Van Der Werf et al., 2008; An-
dela and Van Der Werf, 2014). Therefore, an effective way
of integrating the climate wetness history (i.e., AttentionFire
model) can lead to more accurate predictions of burned-area
spatial-temporal dynamics. https://doi.org/10.5194/gmd-16-869-2023 For the time lags between those dominant climate wetness
variables and fire season burned areas, our results demon-
strated that burned area over NHAF was more modulated by
relatively short-term wetness (VPD during wet-to-dry and
onset of dry season, from September to December), while
SHAF and SHSA burned areas depended more on long-term
wetness (precipitation during wet and wet-to-dry season, De-
cember to March in SHAF and November to April in SHSA)
(Fig. 4g–i). The short-term variations of climate wetness can
directly affect near-surface temperature and moisture avail-
ability, which affect fuel flammability (Littell et al., 2016;
Holden et al., 2018), while the long-term wetness (e.g., dur-
ing rainy season) can affect fuel availability, composition,
and spatial connectivity, which can result in even stronger Furthermore, we found that the emergent functional rela-
tionships between climate wetness and wildfire-burned area
were parabolic (Fig. S3): i.e., enhancement of historical pre-
cipitation or decline of historical VPD (indicating wetter con-
ditions) first increased burned area in more xeric conditions,
then suppressed burned area under more mesic conditions,
consistent with previous findings in subtropical regions (An-
dela and Van Der Werf, 2014; Van Der Werf et al., 2008). The
transition points of these emergent functional relationships
(thresholds at which the relationships reverse) were region https://doi.org/10.5194/gmd-16-869-2023 Geosci. Model Dev., 16, 869–884, 2023 3.3
Possible usage of oceanic index for
long-leading-time predictions While the mean variable im-
portance of OIs was consistently lower than that of local cli- To investigate what drives future burned-area changes
under SSP585, we iteratively surrogated each driver with
its climatology while keeping the other factors the same. Burned-area changing trends in NHAF and SHSA were
mostly affected by VPD changes because removing VPD
inter-annual changes resulted in non-significant burned-area
trends throughout the whole of the NHAF and SHSA re-
gions (Fig. 6a, c). VPD was projected to continuously in- F. Li et al.: AttentionFire_v1.0 F. Li et al.: AttentionFire_v1.0 Figure 5. Performance of AttentionFire burned-area predictions with 1–4 months leading time (short term) and with 5–8 months leading
(long term). MAE is mean absolute error. Four ocean indices which have been widely used for fire prediction over South American and
African regions were considered for long-term forecasting, including Oceanic Niño Index, Atlantic Multidecadal Oscillation index, tropical
Northern Atlantic index, and tropical Southern Atlantic index. Figure 5. Performance of AttentionFire burned-area predictions with 1–4 months leading time (short term) and with 5–8 months leading
(long term). MAE is mean absolute error. Four ocean indices which have been widely used for fire prediction over South American and
African regions were considered for long-term forecasting, including Oceanic Niño Index, Atlantic Multidecadal Oscillation index, tropical
Northern Atlantic index, and tropical Southern Atlantic index. Figure 6. Future burned-area trends under the SSP585 high-emission scenario. (a–c) Spatial distribution of fire season burned-area trends
using drivers with interannual variations; dots in (a)–(c) indicate grid cells with statistically significant changes in the trend. (d–f) Regionally
aggregated burned-area changes with historical mean subtracted. Blue and red lines respectively represent burned-area anomaly in history
and the future; the black line represents the future burned-area trend while removing the interannual variations of the dominant variable. Solid lines represent significant BA trends (p value <0.05), while dashed lines represented non-significant BA trends. Figure 6. Future burned-area trends under the SSP585 high-emission scenario. (a–c) Spatial distribution of fire season burned-area trends
using drivers with interannual variations; dots in (a)–(c) indicate grid cells with statistically significant changes in the trend. (d–f) Regionally
aggregated burned-area changes with historical mean subtracted. Blue and red lines respectively represent burned-area anomaly in history
and the future; the black line represents the future burned-area trend while removing the interannual variations of the dominant variable. Solid lines represent significant BA trends (p value <0.05), while dashed lines represented non-significant BA trends. Fig. S6) through limiting plant growth and fuel availability
(Van Der Werf et al., 2008; Andela and Van Der Werf, 2014;
Andela et al., 2017). For the SHAF, population growth and
climate changes showed stronger influences on burned-area
changes (Andela and Van Der Werf, 2014), while the hetero-
geneity of wildfire responses finally led to a non-significant
trend at the regional scale (Fig. 6). https://doi.org/10.5194/gmd-16-869-2023 https://doi.org/10.5194/gmd-16-869-2023 Geosci. Model Dev., 16, 869–884, 2023 878 3.5
Directions for future research The time-lagged controls of climate on ASA wildfires are
critical for sub-seasonal to seasonal wildfire prediction (Chen
et al., 2020; Andela and Van Der Werf, 2014; Chen et al.,
2011) but remain less well represented due to the complex
interactions among fire, climate, fuel, and human activities. Here, we deployed the interpretable AttentionFire model to
understand and predict fire dynamics in the ASA region. We revealed the dominant, spatially heterogenous, and time-
lagged controls of climate wetness on ASA wildfires. Such
climate wetness importance on ASA wildfires was consis-
tent with previous findings (Andela and Van Der Werf, 2014;
Chen et al., 2011) and was also confirmed by the other three
tree-based ML models (i.e., DT, RF, and GBDT) with vari-
able importance (e.g., precipitation and VPD were regarded
as the top-five most important variables in Fig. S7). How-
ever, differences existed across the model-identified most im-
portant drivers (Fig. 3 versus Fig. S7). The variable impor-
tance of the AttentionFire model was spatiotemporally var-
ied (Fig. 4), while tree-based-model-provided variable im-
portance was constant over the entire dataset. We showed
that the climate wetness was more (less) important in ar-
eas with large (small) burned areas, and its importance also
varied over time (Fig. 4), but the other MLs did not explic-
itly distinguish such differences. Albeit, the higher accuracy
and generally acceptable computation speed of AttentionFire
(Table S2), its memory consumption, and its model training
time could be up to 141 % and 22 times higher than the other
ML models. The implementation of LSTM in the Attention-
Fire model is a serial model instead of a parallel model;
therefore, future work could improve the model efficiency
by deploying some easy-for-parallel-computing time series
prediction frameworks, e.g., temporal convolutional attention
(Lin et al., 2021) and self attention (Mohammadi Farsani and
Pazouki, 2020; Vaswani et al., 2017). F. Li et al.: AttentionFire_v1.0 of climate wetness effects on wildfire dynamics in process-
based and machine-learning-based wildfire prediction mod-
els. al., 2022) and where some extreme wildfires were caused by
persistent drought from wet to dry seasons with multi-month
lags (Taufik et al., 2017; Littell et al., 2016), the Attention-
Fire model could potentially be useful. p
y
With the fully coupled ESMs of CMIP6, we analyzed fu-
ture burned-area changes under high- (SSP585) and low-
emission (SSP126) scenarios in the ASA region. While the
MME mean was considered, substantial uncertainty has been
found across different ESMs in history (Yuan et al., 2022a,
2021; Wu et al., 2020) and the future (Li et al., 2022a; Lauer
et al., 2020). Therefore, further work is needed to narrow
the projection uncertainty of ESMs, e.g., with constraints of
causality (Nowack et al., 2020; Li et al., 2022a) and observa-
tions (Tokarska et al., 2020; Lauer et al., 2020). Meanwhile,
for future projections, although land use and land cover
changes, population growth, and climate and fuel changes
were considered, constant livestock and road density were
adopted due to lack of data. The impacts of livestock and road
density therefore need further exploration with available data
under different future scenarios. In addition, the Attention-
Fire model is currently not coupled with the ESM; therefore,
the feedback among fires, climate, and biomass was ignored. To analyze such feedback, the AttentionFire model needs to
surrogate the original fire module and be coupled with the
ESM (Zhu et al., 2022). F. Li et al.: AttentionFire_v1.0 Our findings highlight
the importance of climate changes for understanding future
burned-area dynamics and motivate for better representation crease due to warming but had different implications over
NHAF and SHSA. Over the relatively fuel-abundant SHSA
region, increased VPD will likely increase burned area (Pear-
son r = 0.64, p value <0.05, Fig. S6) through increasing fuel
dryness and combustibility (Kelley et al., 2019; Chen et al.,
2011; Malhi et al., 2008; Van Der Werf et al., 2008). In
contrast, over the semi-arid savannah-dominated NHAF re-
gion (less fuel, compared with SHSA), higher VPD could
decrease burned area (Pearson r = −0.71, p value <0.05, Geosci. Model Dev., 16, 869–884, 2023 https://doi.org/10.5194/gmd-16-869-2023 879 Geosci. Model Dev., 16, 869–884, 2023 https://doi.org/10.5194/gmd-16-869-2023 References Abatzoglou, J. T. and Kolden, C. A.: Relationships between climate
and macroscale area burned in the western United States, Int. J. Wildland Fire, 22, 1003–1020, 2013. Data availability. We used NCEP-DOE Reanalysis Climate forc-
ings:
https://psl.noaa.gov/data/gridded/data.ncep.reanalysis2.html
(Kanamitsu
et
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and
NOAA
oceanic
index
data:
https://psl.noaa.gov/data/climateindices/list/
(NOAA,
2022). Population
density
data
are
from
https://landscan.ornl.gov/
(Dobson
et
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2000). Road
density
data
are
from
https://www.globio.info/download-grip-dataset
(Meijer
et
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2018). Livestock
density
data
could
be
found
at
https://www.fao.org/dad-is/en/ (Robinson et al., 2014). We used
LUH2 land cover change data: https://luh.umd.edu/data.shtml
(Hurtt
et
al.,
2020). The
grid-cell-level
burned
area
data
are
from
the
Global
Fire
Emissions
Database
https:
//daac.ornl.gov/VEGETATION/guides/fire_emissions_v4.html
(https://doi.org/10.3334/ORNLDAAC/1293,
Randerson
et
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ings:
https://psl.noaa.gov/data/gridded/data.ncep.reanalysis2.html
(Kanamitsu
et
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2002)
and
NOAA
oceanic
index
data:
https://psl.noaa.gov/data/climateindices/list/
(NOAA,
2022). Population
density
data
are
from
https://landscan.ornl.gov/
(Dobson
et
al.,
2000). Road
density
data
are
from
https://www.globio.info/download-grip-dataset
(Meijer
et
al.,
2018). Livestock
density
data
could
be
found
at
https://www.fao.org/dad-is/en/ (Robinson et al., 2014). We used
LUH2 land cover change data: https://luh.umd.edu/data.shtml
(Hurtt
et
al.,
2020). The
grid-cell-level
burned
area
data
are
from
the
Global
Fire
Emissions
Database
https:
//daac.ornl.gov/VEGETATION/guides/fire_emissions_v4.html
(https://doi.org/10.3334/ORNLDAAC/1293,
Randerson
et
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importance: a corrected feature importance measure, Bioinfor-
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Assessing long-term fire risk at local scale by means of de-
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https://doi.org/10.1029/2005JG000133, 2006. Andela, N. and Van Der Werf, G. R.: Recent trends in African fires
driven by cropland expansion and El Niño to La Niña transition,
Nat. Clim. Change, 4, 791–795, 2014. Andela, N., Morton, D. C., Giglio, L., Chen, Y., Van Der Werf,
G., Kasibhatla, P. S., DeFries, R., Collatz, G., Hantson, S., and
Kloster, S.: A human-driven decline in global burned area, Sci-
ence, 356, 1356–1362, 2017. Aragao, L. E. O., Malhi, Y., Barbier, N., Lima, A., Shimabukuro,
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line at: https://doi.org/10.5194/gmd-16-869-2023-supplement. Archibald, S., Roy, D. P., van Wilgen, B. W., and Scholes, R. J.:
What limits fire? An examination of drivers of burnt area in
Southern Africa, Glob. F. Li et al.: AttentionFire_v1.0 Energy under contract no. DEAC02-05CH11231 as part of their Re-
gional and Global Climate Modeling program through the Reducing
Uncertainties in Biogeochemical Interactions through Synthesis and
Computation Scientific Focus Area (RUBISCO SFA) project and as
part of the Energy Exascale Earth System Model (E3SM) project. Code availability. The source code of AttentionFire_v1.0 and all
baseline machine learning models is archived at Zenodo reposi-
tory: https://doi.org/10.5281/zenodo.7416437 (Li et al., 2022b) un-
der Creative Commons Attribution 4.0 International license, with
four zip files: data, data_preparation, model, and example. The
“data” file contains the links to all raw datasets used to drive the
model (e.g., burned areas, climate forcing). The “data_preparation”
file contains the code to preprocess the raw datasets and make
them ready for training and testing of the AttentionFire model. The
“model” file contains the Python code of the AttentionFire model. The “example” file gives a detailed example of how to use the At-
tentionFire model for burned-area predictions. Financial support. This research has been supported by the U.S. Department of Energy (grant no. DEAC02-05CH11231). Review statement. This paper was edited by Mohamed Salim and
reviewed by two anonymous referees. There is also a tutorial file “Data_Model_Tutorial” that contains
descriptions on (1) how to load the raw datasets, (2) how to prepare
the input and output datasets for the ML model, (3) how to initialize
the ML model and run the model, (4) how to train the ML model
and use the trained ML model for predictions, and (5) how to save
and load the model parameters and save the predicted results. 4
Conclusions This study developed an interpretable machine learning
model (AttentionFire_v1.0) for burned-area predictions over
African and South American regions. Compared with ob-
servations and another five widely used machine learning
baseline models, we demonstrated the effectiveness of the
AttentionFire model to capture the magnitude, spatial dis-
tribution, and temporal variation of burned areas. Attention
mechanisms enabled the interpretation of complex but criti-
cal spatial-temporal patterns (Li et al., 2020; Guo et al., 2019;
Liang et al., 2018; Vaswani et al., 2017; Qin et al., 2017), thus
uncovering the black-box relationships in machine learning
models for burned-area predictions. We demonstrated the
spatiotemporally heterogenous and strong time-lagged con-
trols from local climate wetness on burned areas. Further-
more, under the SSP585 high-emission scenario, our results
suggested that the increasing trend in burned area over south-
ern Africa will be dampened, and the declining trend in
burned area over fuel-abundant southern America will re-
verse. This study highlights the importance of the skillful
representation of spatiotemporally heterogenous and strong
time-lagged climate wetness effects on understanding wild-
fire dynamics and developing advanced early fire warning
models. This study focused on wildfire prediction in the ASA re-
gion, and we showed the performance improvement of the
AttentionFire model by representing the time-lagged con-
trols of climate on wildfires. Whether the AttentionFire
model can also outperform other ML models in other regions
may depend on the dependency strength and time lags be-
tween wildfires and climate variables. For example, in North
American boreal forests, lightning was identified as the ma-
jor driver of the interannual variability in burned areas by
influencing the number of ignitions in the dry season (Ve-
raverbeke et al., 2017). In such regions, the AttentionFire
model might not outperform other ML models due to the
lesser dominance of time-lagged controls. In regions like the
western US and India, where wildfires showed time-lagged
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Spatial heterogeneity and infection patterns on epidemic transmission disclosed by a combined contact-dependent dynamics and compartmental model
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Yuhui Zhu
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PLOS ONE PLOS ONE RESEARCH ARTICLE OPEN ACCESS Epidemics, such as COVID-19, have caused significant harm to human society worldwide. A better understanding of epidemic transmission dynamics can contribute to more efficient
prevention and control measures. Compartmental models, which assume homogeneous
mixing of the population, have been widely used in the study of epidemic transmission
dynamics, while agent-based models rely on a network definition for individuals. In this
study, we developed a real-scale contact-dependent dynamic (CDD) model and combined it
with the traditional susceptible-exposed-infectious-recovered (SEIR) compartment model. By considering individual random movement and disease spread, our simulations using the
CDD-SEIR model reveal that the distribution of agent types in the community exhibits spatial
heterogeneity. The estimated basic reproduction number R0 depends on group mobility,
increasing logarithmically in strongly heterogeneous cases and saturating in weakly hetero-
geneous conditions. Notably, R0 is approximately independent of virus virulence when
group mobility is low. We also show that transmission through small amounts of long-term
contact is possible due to short-term contact patterns. The dependence of R0 on environ-
ment and individual movement patterns implies that reduced contact time and vaccination
policies can significantly reduce the virus transmission capacity in situations where the virus
is highly transmissible (i.e., R0 is relatively large). This work provides new insights into how
individual movement patterns affect virus spreading and how to protect people more
efficiently. Citation: Zhu Y, Shen R, Dong H, Wang W (2023)
Spatial heterogeneity and infection patterns on
epidemic transmission disclosed by a combined
contact-dependent dynamics and compartmental
model. PLoS ONE 18(6): e0286558. https://doi. org/10.1371/journal.pone.0286558 Editor: Junyuan Yang, Shanxi University, CHINA
Received: January 11, 2023
Accepted: May 18, 2023
Published: June 13, 2023 Peer Review History: PLOS recognizes the
benefits of transparency in the peer review
process; therefore, we enable the publication of
all of the content of peer review and author
responses alongside final, published articles. The
editorial history of this article is available here:
https://doi.org/10.1371/journal.pone.0286558 Copyright: © 2023 Zhu et al. This is an open access
article distributed under the terms of the Creative
Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in
any medium, provided the original author and
source are credited. Data Availability Statement: All relevant data are
accessible through this DOI:(10.5061/dryad. qrfj6q5mp). Spatial heterogeneity and infection patterns
on epidemic transmission disclosed by a
combined contact-dependent dynamics and
compartmental model Youyuan Zhu1,2,3, Ruizhe Shen1,2,3, Hao DongID1,3,4*, Wei Wang2,3,5* 1 Kuang Yaming Honors School, Nanjing University, Nanjing, China, 2 Collaborative Innovation Center of
Advanced Microstructures, National Laboratory of Solid State Microstructure, Nanjing, China, 3 Department
of Physics, Nanjing University, Nanjing, China, 4 State Key Laboratory of Analytical Chemistry for Life
Science, Nanjing University, Nanjing, China, 5 Institute for Brain Sciences, Nanjing University, Nanjing, China a1111111111
a1111111111
a1111111111
a1111111111
a1111111111 * donghao@nju.edu.cn (HD); wangwei@nju.edu.cn (WW) Introduction The outbreak of COVID-19 brought a profound disaster to human civilization worldwide,
which has infected more than 600 million people and killed more than 6 million people by
October 2022, and the virus is still in constant mutation [1, 2]. From the original strain to the Funding: This work was supported by the National
Natural Science Foundation of China (Grant Nos. PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 1 / 16 PLOS ONE The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading 12090052, 21833002), the National Key R&D
Program of China (2021ZD201302), and the
“Fundamental Research Funds for the Central
Universities” (021514380018). Parts of the
calculations were performed using computational
resources on an IBM Blade cluster system from the
High-Performance Computing Center (HPCC) of
Nanjing University. The funders had no role in
study design, data collection and analysis, decision
to publish, or preparation of the manuscript. 12090052, 21833002), the National Key R&D
Program of China (2021ZD201302), and the
“Fundamental Research Funds for the Central
Universities” (021514380018). Parts of the
calculations were performed using computational
resources on an IBM Blade cluster system from the
High-Performance Computing Center (HPCC) of
Nanjing University. The funders had no role in
study design, data collection and analysis, decision
to publish, or preparation of the manuscript. Alpha, Delta, and Omicron variants, while the virulence of the virus decreased, the infectivity
gradually increased. As an important indicator to characterize infectious diseases, the basic
reproduction number (R0) has increased from 2.79 in the original strain to 9.5 in the Omicron
variant [3–5], bringing new challenges to epidemic prevention and control. In the theoretical models of studying the transmission of infectious disease, homogeneous
mixing [6] is a classic assumption of the traditional compartmental model [7]. This approxi-
mation simplifies the model and thus can help to obtain epidemic transmission trends more
efficiently. However, in real situations, heterogeneity, including differences between individu-
als [8–10] and differences in spatial distribution [11–14], is also an important factor in epi-
demic transmission that should be considered [8–14]. Competing interests: The authors have declared
that no competing interests exist. Competing interests: The authors have declared
that no competing interests exist. In recent years, agent-based models (ABMs) that focus on individual behavior (even con-
sidering the corresponding movement characteristics) have received increasing attention [15,
16]. This model examines the overall behavior and trends by modeling many individuals
(called agents). Introduction In ABM, individuals are made autonomous and adaptive by defining them to
choose behaviors based on their own decision rules (often derived from limited information in
their immediate surroundings), and the decision rules could vary from agent to agent. Agents
are social and interdependent. If an agent performs a certain action, other agents may be influ-
enced and respond in some way. A central concept of ABM is that the overall characteristics of
a group are not simply the sum of all individual characteristics in the group but are the result
of interactions guided by individual characteristics. Interactions among individuals often
result in a society with a different overall identity from that of the individuals. In the case of
epidemic transmission, the spatial movement of individuals and the influence of spatial hetero-
geneity on the spread of an epidemic are important factors that cannot be ignored. Adopting
the idea of ABM to study may be an appropriate idea [12, 14, 17–23]. In some ABMs, the location and interdependence between individuals are determined by a
predefined social network (e.g., building a small-world network to simulate the social relation-
ships of agents), and the spread of epidemics is demonstrated through the evolution of the net-
work [24, 25]. Complex network models can better introduce heterogeneity among In some ABMs, the location and interdependence between individuals are determined by a
predefined social network (e.g., building a small-world network to simulate the social relation-
ships of agents), and the spread of epidemics is demonstrated through the evolution of the net-
work [24, 25]. Complex network models can better introduce heterogeneity among
individuals but lack the spatial movement of individuals [19]. In addition, there are individual-
based motion models that rely not on defined social networks but only on individual motion
[26, 27]. These models suggest that characteristic quantities of motion, such as the velocity of
individuals, can play an important role in the spread of the epidemic [19, 20]. However, these
models often simplify factors related to epidemic transmission because of computational effi-
ciency [12, 14, 17–22]. For example, the motion and the motion features of an individual at a
real time-space scale are not fully considered. [
,
]
p
g
y
g
individuals but lack the spatial movement of individuals [19]. PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 Introduction In addition, there are individual-
based motion models that rely not on defined social networks but only on individual motion
[26, 27]. These models suggest that characteristic quantities of motion, such as the velocity of
individuals, can play an important role in the spread of the epidemic [19, 20]. However, these
models often simplify factors related to epidemic transmission because of computational effi-
ciency [12, 14, 17–22]. For example, the motion and the motion features of an individual at a
real time-space scale are not fully considered. In this work, we designed a contact-dependent dynamics (CDD) model based on the char-
acteristics of epidemic transmission to mimic the motion of the population within a commu-
nity. Our model consisted of alternating individual motions and contact between individuals
(thus changing the direction of motion) in an ensemble of individuals. We combined this
CDD model (to describe population movement) with the commonly used epidemic transmis-
sion model, the susceptible-exposed-infectious-recovered (SEIR) model (to describe infection
between individuals), to study the characteristics of epidemic transmission by studying the
movement of individuals at real time and space scales and epidemic transmission. We found
that the CDD model has a larger proportion of short-term contacts as the velocity of agents
decreases. To characterize the epidemic transmission capacity, we calculated the basic repro-
duction number R0 of the model. The calculated R0 with the CDD-SEIR model is nearly inde-
pendent of the virulence of the virus and much less than that calculated by mean-field
equations [19], which is attributed to the short exposure time ω of the CDD model when the
velocity of all individuals in the population is less than a threshold value. This corroborates the PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 2 / 16 PLOS ONE The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading effectiveness of prevention and control measures in the presence of highly infectious strains. Moreover, we found that R0 shows two stages of variation with the spatial heterogeneity of the
system: that is, in the strong heterogeneity range, R0 is proportional to the logarithm of veloc-
ity, which is different from mean-field theory, and in the weak heterogeneity range, R0 reaches
saturation. The above results provide us with a new understanding of the relationship between
epidemic transmission and individual movement. This paper is organized as follows: we first propose a modified SEIR model to incorporate
individual mobility. Introduction Then, we introduce the CDD model designed to characterize the stochas-
tic motion of individuals within a community. Next, we describe the CDD-SEIR model, com-
bining real-scale stochastic individual motion with epidemic transmission. To characterize the
agent dynamics, the exposure time ω and mean free time τ will be discussed. Meanwhile, we
show the characteristics of epidemic spreading described by this CDD-SEIR model, as well as
the calculated basic reproduction number R0 and the spatial heterogeneity z, obtaining the
effect of motion parameters (e.g., velocity) on these properties. We will then discuss the bipar-
tite variation of R0 with the strength of spatial heterogeneity. Notably, R0 is nearly independent
of the virulence of the virus when all individuals move at less than a threshold value and spatial
heterogeneity is sufficiently strong. R0 is much less than that calculated by mean-field equa-
tions due to the short exposure time caused by the CDD model. Therefore, an in-depth under-
standing of heterogeneity is important for an accurate understanding of epidemic
transmission, e.g., prevention and control measures are still effective when the virulent strain
is highly infectious due to the presence of spatial heterogeneity. The short-term contact model transmission, e.g., prevention and control measures are still effective when the virulent strain
is highly infectious due to the presence of spatial heterogeneity. The short-term contact model
can greatly suppress the epidemic when combined with control measures. PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 The SEIR model The SEIR model is one of the compartmental models that is commonly used to predict epi-
demic transmission [28]. Here, S, E, I and R stand for susceptible, exposed, infectious, and
recovered, respectively, which are the four categories of people in epidemic transmission. The
sum of these four is the population within the community. In the SEIR model, susceptible peo-
ple who become infected come through exposed, infectious, and recovered compartments suc-
cessively, as shown in Fig 1A. The latent period was considered when individuals had been
infected but were not yet infectious themselves. COVID-19 has been reported to be transmitted mainly by proximity contact [29]. To intro-
duce the mobility of individuals into the study of epidemic transmission, we proposed a modi-
fied SEIR model. According to the definition of the SEIR model, the agents here fall into four
categories: susceptible, PS, healthy individuals who may be infected; exposed, PE, individuals
who have been infected but do not cause infection after an incubation period; infectious, PI,
individuals who have been infected and can transmit the virus to PS; and recovered, PR, indi-
viduals who have been cured or immunized and are thus removed from the transmission
process. In the modified SEIR model (Fig 1B), the PS has a probability pSE to convert to PE when the
distance r between the susceptible PS and the infected PI is less than the airborne transmission
distance d. The latent period for COVID-19 was estimated to be approximately 7 days in the
literature [2, 30]. Accordingly, we set the latent period of PE between 4 and 10 days. Specifi-
cally, within 0–4 days, PE only acts as the virus carrier and does not have the ability to infect
others; within 4–10 days, PE can transform with the probability pEI (Eq 1, where T is in the
unit day) to PI, which can transmit the virus; otherwise, it remains as PE. After 10 days, PE that
still does not convert to PI is classified as PR. Meanwhile, PI can be transformed into PR with 3 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 PLOS ONE The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading Fig 1. The classic SEIR model and the modified SEIR model. (a) The classic SEIR model. Green, black, red, and blue
squares represent susceptible, exposed, infectious, and recovered compartments, respectively. The SEIR model Susceptible individuals
in the system successively go through these four stages. Transitions between adjacent stages occur according to certain
probabilities. (b) The modified SEIR model. When the distance between the susceptible PS and PI is within the
airborne transmission distance d, PS can be infected by PI and transformed into PE at the probability pSE. The latent
period is within 4–10 days. During this period, PE converts with probability pEI to infectious PI that can transmit the
virus. Any PE that remained unconverted to a PI after 10 days is classified as a recover PR. Meanwhile, PI could also be
cured with probability pIR and transformed into PR. Fig 1. The classic SEIR model and the modified SEIR model. (a) The classic SEIR model. Green, black, red, and blu
squares represent susceptible, exposed, infectious, and recovered compartments, respectively. Susceptible individuals
in the system successively go through these four stages. Transitions between adjacent stages occur according to certain
probabilities. (b) The modified SEIR model. When the distance between the susceptible PS and PI is within the
airborne transmission distance d, PS can be infected by PI and transformed into PE at the probability pSE. The latent
period is within 4–10 days. During this period, PE converts with probability pEI to infectious PI that can transmit the
virus. Any PE that remained unconverted to a PI after 10 days is classified as a recover PR. Meanwhile, PI could also be
cured with probability pIR and transformed into PR. Fig 1. The classic SEIR model and the modified SEIR model. (a) The classic SEIR model. Green, black, red, and blue
squares represent susceptible, exposed, infectious, and recovered compartments, respectively. Susceptible individuals
in the system successively go through these four stages. Transitions between adjacent stages occur according to certain
probabilities. (b) The modified SEIR model. When the distance between the susceptible PS and PI is within the
airborne transmission distance d, PS can be infected by PI and transformed into PE at the probability pSE. The latent
period is within 4–10 days. During this period, PE converts with probability pEI to infectious PI that can transmit the
virus. Any PE that remained unconverted to a PI after 10 days is classified as a recover PR. Meanwhile, PI could also be
cured with probability pIR and transformed into PR. https://doi.org/10.1371/journal.pone.0286558.g001 https://doi.org/10.1371/journal.pone.0286558.g001 https://doi.org/10.1371/journal.pone.0286558.g001 the probability pIR. the probability pIR. The SEIR model pEI ¼
0
T < 4
pEI;Day7∗
2
1 þ e3ðT 7Þ
4 T < 10
ð1Þ
8
<
: ð1Þ The conversion probability from the exposed agent (PE) to the infectious agent (PI) pEI is
reduced with the increase in Day T from latency, as shown in Eq 1. pEI,Day7 is the conversion
probability of the 7th day of the latent period, and its value is shown in Table 1. The CDD model In this case, the velocity is
randomly generated by the normal distribution, and the direction of motion is randomly reas-
signed by the normal distribution within the range of ±45 degrees from the original motion
direction. To represent the real-scale motion, the units for length and time used in this work
were in meters and seconds, respectively. We set rc to 2 m and used the time interval Δt of 1 s. when there are other agents within the distance rc around i (Eq 2). In this case, the velocity is
randomly generated by the normal distribution, and the direction of motion is randomly reas-
signed by the normal distribution within the range of ±45 degrees from the original motion
direction. To represent the real-scale motion, the units for length and time used in this work
were in meters and seconds, respectively. We set rc to 2 m and used the time interval Δt of 1 s. viðt þ DtÞ; yiðt þ DtÞ ¼
viðtÞ; yiðtÞ
no contact
vrnd; yiðtÞ þ yrnd
with contact
ð2Þ
( ð2Þ where vi(t) and θi(t) denote the velocity and direction of particle i at time t, respectively. vrnd is
the reassigned velocity that satisfies a normal distribution with a mean of v and a standard
deviation of 1
3 v. θrnd is the change in the reassigned direction (in the range of -45˚ to 45˚),
which satisfies a normal distribution with a mean of 0˚ and a standard deviation of 15˚. The
distributions of vrnd and θrnd are shown in Fig 2B. Notably, this CDD model has some similarities to the stochastic rotation dynamics (SRD)
model [31, 32], which has been widely used to characterize fluid flow. The main difference
between the two models is that in the CDD model, the individuals change direction and veloc-
ity of motion when the distance between two individuals is less than a certain range, rather
than waiting until they are in full contact and collide. Such a model setup provides the possibil-
ity to study the ability of the virus itself to spread in space for the spread of the epidemic and
for social distancing interventions. The CDD model We introduce the CDD model to characterize the motion of agents. In CDD, the agents are
described by points. Each agent i is assigned a velocity vi following a normal distribution with
a mean of v, a standard deviation of 1
3 v, and a direction θi randomly generated from a uniform
distribution. The motion of the agents consists of alternating individual motions and contact
between individuals (thus changing the motion) in an ensemble of individuals. When there are no other agents within the distance rc around agent i, the latter adopts uni-
form linear motion, and the evolution of its position is determined only by its velocity; other-
wise, the direction of motion and the velocity of i will be regenerated, as shown in Fig 2A, PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 4 / 16 PLOS ONE The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading Table 1. The different mobility and propagation parameters tested in this model. We selected 9 different velocities
v and 5 different values for the remaining parameters, including the initial proportion of PI nI,0, the airborne transmis-
sion distance d, the infection probability pSE, the conversion probability of Day 7 pEI,Day7 from PE to PI, and the recov-
ery probability pIR. Those marked with * are the default values. In each change of one parameter, the default values
were used for the other parameters. For each set of parameter combinations, we calculated 5 random motion trajecto-
ries of agents at 40 days length separately using the CDD model and postprocessed each trajectory using the SEIR
model to obtain 5 independent modes of transmission. The final reported results were extracted from the average of
5*5 = 25 trajectories. Parameters
Values
v (10-4m/s)
1, 2, 5, 10, 20, 50, 100, 200, 500
nI,0
1%*, 3%, 5%, 7%, 9%
d (m)
1, 2*, 3, 4, 5
pSE (10−4)
0.5, 1.0, 1.5, 2.0*, 2.5
pEI,Day7 (10−6)
1, 2*, 3, 4, 5
pIR (10−6)
1, 2*, 3, 4, 5
https://doi.org/10.1371/journal.pone.0286558.t001 Parameters
Values
v (10-4m/s)
1, 2, 5, 10, 20, 50, 100, 200, 500
nI,0
1%*, 3%, 5%, 7%, 9%
d (m)
1, 2*, 3, 4, 5
pSE (10−4)
0.5, 1.0, 1.5, 2.0*, 2.5
pEI,Day7 (10−6)
1, 2*, 3, 4, 5
pIR (10−6)
1, 2*, 3, 4, 5
https://doi.org/10.1371/journal.pone.0286558.t001 https://doi.org/10.1371/journal.pone.0286558.t001 when there are other agents within the distance rc around i (Eq 2). The CDD-SEIR model We combined the CDD model (to describe population movement) with the modified SEIR
model (to describe infection between individuals) to simulate epidemic transmission at real
time and space scales. As shown in Fig 3A, a community represented by a periodic square box with side length L
has N individuals. The population is composed of susceptible PS, exposed PE, infectious PI and
recovered PR. The number of each type is NS, NE, NI and NR, respectively, where N = NS + NE
+ NI + NR. In subsequent work, we take the early stage of the epidemic as the initial condition
of the simulations, put only a small amount of PI in the system and set the rest of the particles
as PS. It should be noted that other factors, such as repeated infections, vaccination, etc., have
potential influence on the transmission mechanism, and can be easily added to the current 5 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading PLOS ONE Fig 2. Schematic diagram of the CDD model. (a) Stochastic motion process characterized by the CDD model. The
upper panel represents that the direction of motion of an agent varies within [-45˚, 45˚] of the original direction when
there are other agents within rc. The lower panel gives an example of the change in motion state of two individuals
before and after contact: i is the central agent, and the blue circle represents its contact range of radius rc. Individuals i
and j fall in contact when their distance is less than rc, and the agents are in a uniform linear motion before contact. The red, green, and black solid vectors represent the velocities of i and j and the relative velocity j relative to i,
respectively. The gray dashed line represents the trajectory of j relative to i. (b) The reassigned velocity vrnd that
satisfies a normal distribution with a mean of v and a standard deviation of 1
3 v. θrnd is the change in the reassigned
direction (in the range of -45˚ to 45˚), which satisfies a normal distribution with a mean of 0˚ and a standard deviation
of 15˚. Fig 2. Schematic diagram of the CDD model. (a) Stochastic motion process characterized by the CDD model. The Fig 2. Schematic diagram of the CDD model. (a) Stochastic motion process characterized by the CDD model. The CDD-SEIR model The
upper panel represents that the direction of motion of an agent varies within [-45˚, 45˚] of the original direction when
there are other agents within rc. The lower panel gives an example of the change in motion state of two individuals
before and after contact: i is the central agent, and the blue circle represents its contact range of radius rc. Individuals i
and j fall in contact when their distance is less than rc, and the agents are in a uniform linear motion before contact. The red, green, and black solid vectors represent the velocities of i and j and the relative velocity j relative to i,
respectively. The gray dashed line represents the trajectory of j relative to i. (b) The reassigned velocity vrnd that
satisfies a normal distribution with a mean of v and a standard deviation of 1
3 v. θrnd is the change in the reassigned
direction (in the range of -45˚ to 45˚), which satisfies a normal distribution with a mean of 0˚ and a standard deviation
of 15˚. Fig 3. System setup of the CDD-SEIR model. (a) Schematic diagram of a representative system obtained from the
simulations. The system contains 200 particles with a box length of 100
ffiffiffi
2
p
m and density of (100 m2)-1. PS, PE, PI and
PR are represented by green, black, red and blue particles, respectively. (b) The profiles of the temporal population of
PS (in green), PE (in black), PI (in red) and PR (in blue) during epidemic transmission. (c) Representative snapshots of
the infection within a community at days 0, 10, 20, 30, and 40 are shown. https://doi org/10 1371/journal pone 0286558 g003 Fig 3. System setup of the CDD-SEIR model. (a) Schematic diagram of a representative system obtained from the
simulations. The system contains 200 particles with a box length of 100
ffiffiffi
2
p
m and density of (100 m2)-1. PS, PE, PI and
PR are represented by green, black, red and blue particles, respectively. (b) The profiles of the temporal population of
PS (in green), PE (in black), PI (in red) and PR (in blue) during epidemic transmission. (c) Representative snapshots of
the infection within a community at days 0, 10, 20, 30, and 40 are shown. The CDD-SEIR model https://doi.org/10.1371/journal.pone.0286558.g003 https://doi.org/10.1371/journal.pone.0286558.g003 6 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 PLOS ONE The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading model (by adding types of agents). Meanwhile, factors such as death can also be implemented
in the current model by removing particles from the system or eliminating their movement. The motion of agents is driven by the CDD model, and the spread of the epidemic is deter-
mined by the modified SEIR model. Therefore, we obtained the temporal evolution of the
agents’ populations (Fig 3B) and the dynamic progress of the infection spreading (Fig 3C). We can study the factors of epidemic spreading from the perspective of both spatial agent
dynamics and epidemic dynamics. Comparing the results obtained from the present CDD-SEIR model with the real epidemic
data is a way to test the model’s reliability. However, limited by the current model setup, such
a direct comparison is not easy: first, we used 1 s as the integration step of the equation of
motion to not miss any of the exposure events, and thus currently, the model can only handle
a few hundred agents in a few tens of days within an affordable computational cost; second,
for a system containing hundreds of particles, a single initial infected agent already corre-
sponds to a very high initial infection rate (e.g., 1 in 200 agents corresponds to an initial infec-
tion rate of 2.5%), while the ratio in the real situation is often less than 1 in 10,000. Therefore,
the present work lies mainly in providing a simple model with clear physical meaning to study
the dynamics of epidemic transmission. Parameter selection of the CDD-SEIR model Considering the computational efficiency, in the simulations, we used the following parame-
ters for the CDD model: number of particles N = 200, density of particles ρ = (100 m2)-1, and
contact distance rc = 2 m. These parameters remained intact in the simulations. Forty-day tra-
jectories were accumulated separately under the different combinations of parameters. Based on the case of COVID-19 propagation [2], we set the default values of some propaga-
tion parameters in the SEIR model to be close to the real situation. For example, we assumed
the airborne transmission distance d = 2 m. This is considered a safe social distance to prevent
the spread of the virus [33]. In addition, we set pEI,Day7 = pIR = 2*10−6, which determines the
duration of the incubation and disease periods, and assumed that pSE = 2*10−4, which deter-
mines R0 to be close to the real situation based on the reported data [30, 34, 35]. Meanwhile, we carefully examined the effects of the rest of the propagation parameters in
the combined CDD-SEIR model, including the mobility v, the initial proportion of PI nI,0, the
airborne transmission distance d, the infection probability pSE, the conversion probability of
Day 7 from the exposed agent (PE) to the infectious agent (PI) pEI,Day7, and the recovery proba-
bility pIR. By changing the value of the parameters at a time (keeping the other parameters at
their default values), we systematically studied the effect of that parameter and the combina-
tion of different parameters on the spread of the epidemic. The values of the parameters are
shown in Table 1. Agent dynamics and contact pattern To investigate the effect of contact-dependent dynamics, we estimate the exposure time ω (the
time that two particles stay in contact) and the mean free time τ (the time interval between two
consecutive contacts) in the CDD model. By counting the time that particles i and j remain in contact in trajectories, we can estimate
the exposure time ω of different velocities in the CDD model. As shown in Fig 4A, the average
of ω is inversely proportional to v, while the median decreases slightly as v increases. This
means that the agents do not intend to stay together for a long time as v decreases. We classify
contacts into short-term contact (ω < 0.5*rc/v), medium-term contact (0.5*rc /v ω < 4*rc 7 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading PLOS ONE Fig 4. Contact pattern of the CDD model. (a) The mean (blue) and median (red) of the contact time ω influenced by
the velocity v of agents in the CDD model. (b) The percentages of ω in different time ranges for the velocity v of agents
in the CDD model. The blue line represents ω < 0.5*rc /v, the red line represents 0.5*rc /v ω < 4*rc /v, and the green
line represents ω 4*rc/v. (c) The mean free time τ in the CDD model (solid line) and the CDD-NC model (dashed
line) for different velocities. Fig 4. Contact pattern of the CDD model. (a) The mean (blue) and median (red) of the contact time ω influenced by
the velocity v of agents in the CDD model. (b) The percentages of ω in different time ranges for the velocity v of agents
in the CDD model. The blue line represents ω < 0.5*rc /v, the red line represents 0.5*rc /v ω < 4*rc /v, and the green
line represents ω 4*rc/v. (c) The mean free time τ in the CDD model (solid line) and the CDD-NC model (dashed
line) for different velocities. https://doi.org/10.1371/journal.pone.0286558.g004 https://doi.org/10.1371/journal.pone.0286558.g004 https://doi.org/10.1371/journal.pone.0286558.g004 /v) and long-term contact (ω 4*rc/v) by the value of ω (Fig 4B). The results show that the
CDD model has a larger proportion of short-term contacts as the velocity of agents decreases. Agent dynamics and contact pattern In addition, by dividing the total trajectory time by the average number of particle contacts
obtained from the statistics of the CDD model, we can estimate the mean free time τ. As a con-
trol, we also set up a contact-dependent dynamics with no contact (CDD-NC) model (S1 and
S2 Figs in S1 File, please refer to the S1 File for more details), in which the velocities and
directions of the individuals follow the same distribution in CDD and the motion does not
change when two particles are in contact, as in the no contact case of Eq 2. We further esti-
mated the mean free time τ (Eq 3) of the CDD-NC model. t ¼
1
2rc v0r
ð3Þ ð3Þ where rc is the contact distance, as defined before, v0 is the average relative velocity and was set
as v0 ¼ 1:35v, and ρ is the density of the system. With respect to τ, we found that there was no
difference between the two models (Fig 4C). Therefore, the CDD model does not change the frequency of contact but only makes the
contact pattern of individuals more inclined to short-term contact as the velocity of agents
decreases. In the following, we found that this contact pattern caused strong spatial heteroge-
neity and determined the transmission pattern at low velocity. PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 Epidemic dynamics and the basic reproduction number R0 We examined the effect of five propagation parameters, including nI,0, d, pSE, pEI,Day7 and pIR
on epidemic evolution. For example, different pSE agents show different patterns of epidemic
spread as shown in Fig 5A and 5B. To reasonably characterize the epidemic transmission capacity obtained from the current
CDD-SEIR model simulations, we defined the basic reproduction number R0. Traditional
models commonly use the basic reproduction number R0 to indicate how contagious an infec-
tious disease is. R0 is the number of secondary infections an infectious PI can cause in a fully
susceptible environment [36]. We calculated the basic reproduction number R0 by the defini-
tion of R0 in ordinary differential equations. Similar to SEIR model, dNS
dt ¼ b NI
N NS
ð4Þ ð4Þ 8 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 PLOS ONE The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading Fig 5. Infection spreading in different pSE and the basic reproduction number R0 influenced by pSE and the velocity v of
agents in the system. The dynamic progress of the infection spreading when pSE = 1.0*10−4 (a) and pSE = 2.0*10−4 (b). Snapshots of Days 0, 20, and 40 are shown in order. (c) The influence of pSE and v of agents on R0 in the system. The points
represent the calculated data, and the solid line represents the fitting curve to eliminate noise. https://doi.org/10.1371/journal.pone.0286558.g005 Fig 5. Infection spreading in different pSE and the basic reproduction number R0 influenced by pSE and the velocity v of
agents in the system. The dynamic progress of the infection spreading when pSE = 1.0*10−4 (a) and pSE = 2.0*10−4 (b). Snapshots of Days 0, 20, and 40 are shown in order. (c) The influence of pSE and v of agents on R0 in the system. The points
represent the calculated data, and the solid line represents the fitting curve to eliminate noise. https://doi.org/10.1371/journal.pone.0286558.g005 dNR
dt ¼ gNI
ð5Þ ð5Þ where NS, NI, NR and N represent the susceptible, infectious, recovered and total populations
of agents, and β and γ represent the transmission rate and recovery rate (which equals to pIR
in our model), respectively. PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 Epidemic dynamics and the basic reproduction number R0 Then R0 can be calculated by R0 ¼ b
g
ð6Þ ð6Þ We calculated the basic reproduction number R0 in the CDD-SEIR model by R0 ¼ b
pIR
¼
N
pIRdt
dNS
NSNI
N
pIRDt < DNS
NSNI
>
ð7Þ ð7Þ where ΔNS represents the variation of NS in the integration time step Δt (please refer to the
S1 File for more details). Fig 5C shows how R0 is influenced by pSE and the velocity v of agents in the system. In addi-
tion to R0, we also counted the peak proportion of infections in the system, nI,max, and the
infection rate p of the system when the epidemic had developed for 40 days to characterize
spreading (S4 Fig in S1 File). We examined the effect of five propagation parameters, including nI,0, d, pSE, pEI,Day7, and
pIR, on R0 when the agents in the system had a specific mobility. As shown in Fig 6A, except
for the parameters nI,0 and pEI,Day7, R0 shows a significant dependence on the other three
parameters. Specifically, the ability of the virus to spread/infect is positively correlated with d,
and pSE and negatively correlated with pIR. This observation is consistent with our common
perception: the larger the infection range (d), the more virulent the virus; the lower the recov-
ery rate, the higher the severity of the outbreak. Meanwhile, it should be noted that these three
parameters also showed different effects: in the speed range studied in the current work, R0
shows a clear dependence on the infection range d, i.e., a larger d led to a larger R0; in contrast,
R0 is not significantly affected by different pSE and pIR when the system is under low mobility
and only reflects notable differences when the agents are in high mobility. We fit the curves for each of the two phases of R0 linearly and define the critical velocity vc
of R0 at which R0 reaches 90% of the saturated reproduction number (please refer to the PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 9 / 16 PLOS ONE The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading Fig 6. The basic reproduction number R0 (a), the heterogeneity variable z (b) and the critical velocity vc of R0 and z (c) influenced by
five parameters (nI,0, d, pSE, pEI,Day7, and pIR) and the velocity v of agents in the system. Epidemic dynamics and the basic reproduction number R0 In each plot, the results corresponding to the
five values used for the parameter that changed are represented by five different colored curves. The points represent the calculated
data, and the solid line represents the fitting curve to eliminate noise. The horizontal coordinates in (a) and (b) are on a logarithmic
scale. Fig 6. The basic reproduction number R0 (a), the heterogeneity variable z (b) and the critical velocity vc of R0 and z (c) influenced by
five parameters (nI,0, d, pSE, pEI,Day7, and pIR) and the velocity v of agents in the system. In each plot, the results corresponding to the
five values used for the parameter that changed are represented by five different colored curves. The points represent the calculated
data, and the solid line represents the fitting curve to eliminate noise. The horizontal coordinates in (a) and (b) are on a logarithmic
scale. https://doi.org/10.1371/journal.pone.0286558.g006 S1 File for more details). The calculations show that when the mobility of the system is small
(i.e., the individual’s v is less than vc), R0 is influenced almost exclusively by d, independent of
other propagation parameters. When v > vc, the system is close to the scenario obtained from
the classical epidemic model assuming uniform mixing [6]. For example, in classical models,
an infectious agent can come into contact with susceptible individuals at a probability propor-
tional to NS. Thus, the mobility of the infectious is not directly reflected in the process of virus
transmission. However, when v < vc, this is not the case. Epidemics transmitted by contact are
closely related to the type of individuals who can be reached locally, the so-called spatial het-
erogeneity [12, 14]. PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 Heterogeneity variable z and spatial heterogeneity Fig 6A clearly shows that the epidemic varies linearly with v when v < vc, indicating that het-
erogeneity significantly influences the spread of the epidemic; in contrast, the spread of the
epidemic is no longer dependent on the variation of v when v > vc [20]. Therefore, it is neces-
sary to explore the factors influencing vc. As shown in Fig 6C, we found that vc of R0 is tightly
associated with certain propagation parameters. Specifically, vc is positively correlated with
pSE, and nearly independent of d, initial nI,0, pEI,Day7, and pIR. It should be noted that the inde-
pendence between vc and the initial nI,0 indicates that the epidemic is determined by its inher-
ent properties. On the other hand, pEI, Day7 and pIR do not affect the distribution of PS around
PI, which is associated with heterogeneity and thus vc. Fig 6A clearly shows that the epidemic varies linearly with v when v < vc, indicating that het-
erogeneity significantly influences the spread of the epidemic; in contrast, the spread of the
epidemic is no longer dependent on the variation of v when v > vc [20]. Therefore, it is neces-
sary to explore the factors influencing vc. As shown in Fig 6C, we found that vc of R0 is tightly
associated with certain propagation parameters. Specifically, vc is positively correlated with pSE, and nearly independent of d, initial nI,0, pEI,Day7, and pIR. It should be noted that the inde-
pendence between vc and the initial nI,0 indicates that the epidemic is determined by its inher-
ent properties. On the other hand, pEI, Day7 and pIR do not affect the distribution of PS around
PI, which is associated with heterogeneity and thus vc. Apparently, vc indicates the boundary between heterogeneity and homogeneity. To further
elucidate the above trend, we calculated the relative neighbor concentration RNCS of the sys-
tem for characterizing spatial heterogeneity: RNCS was defined as the average of the ratio of
the local concentration of PS within the infection range d of a single PI in the system to the
global concentration of PS in the whole system. The heterogeneity variable z was defined as 1- PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 10 / 16 PLOS ONE The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading RNCS (Eq 8). Heterogeneity variable z and spatial heterogeneity ζ ¼ 1 RNCS ¼ 1 < N0
S=N0
NS=N >
ð8Þ ð8Þ Here, N´S represents the total number of PS within radius d of a PI, N´ represents the total
number of all individuals within radius d of this PI, and the ratio of these two corresponds to
the local concentration of PS; NS/N represents the global concentration of PS (please refer to
the S1 File for more details). As shown in Fig 6B, z gradually converges to 0 with increasing v, indicating that the system
gradually transitions from a strongly heterogeneous system to a weakly heterogeneous system
(therefore satisfying the assumption of homogeneous mixing). We can also define the critical
velocity vc of z at which z reaches 0.1(RNCS reaches 0.9). As shown in Fig 6C, the saturation
velocity of z is comparable to that of R0, and both are positively correlated with pSE, and less
correlated with d, initial nI,0, pEI,Day7, and pIR. This is easy to understand. When pSE increases,
the number of PS decreases while PI hardly reaches new susceptible particles, so the spatial het-
erogeneity is further enhanced. The spatial heterogeneity of agents other than PS is not
strongly associated with PS. When d increases, the distribution of agents within d (local distri-
bution) is closer to the distribution of agents within the whole system (global distribution), so
the spatial heterogeneity decreases. However, this effect seems not significant to vc. Therefore,
the trend of the two phases of R0 under different velocity conditions is caused by spatial
heterogeneity. Fig 6 shows that the variation in R0 with velocity shows two different phases, with the
boundary of vc. We further investigated the relationship between the maximum slope k
obtained by fitting the data before vc and the saturated reproduction number Rs after vc and
other propagation parameters (Fig 7). The two indexes, k and RS, have consistent variability,
i.e., they are both positively correlated with d and pSE, negatively correlated with pIR, and inde-
pendent of the initial nI,0 and pEI,Day7. For RS, we further find that the data can be approxi-
mately proportional to d2pSE/pIR. In this case, the result obtained from the CDD model is close
to that of the mean field method. Notably, the results obtained from the CDD-SEIR model show that R0 is linearly related to
log(v). https://doi.org/10.1371/journal.pone.0286558.g007 PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 Effect of the CDD model on R0 As shown in Fig 6, when v < vc, the basic reproduction number R0 is nearly independent of
the virulence pSE. In the work of the mean field models [19], R0 ¼ ð1 e o=tTÞvs0rtI, where ω
is the exposure time, τT is the characteristic time of infection, 1 e o=tT represents the proba-
bility of infection in each contact, σ0 is the scattering cross section of the agents, ρ is the den-
sity, and τI is the transition time from infectious to recovered. When the mobility of the agents is low, ω is much larger than τT, and therefore, infection
must occur in each contact, resulting in R0 that is independent of viral virulence. From this
perspective, the results obtained by the CDD-SEIR model are consistent with those obtained
by the mean-field model. However, there are naturally obvious differences between the two. Spatial heterogeneity caused by the contact model of the CDD model plays an important role
in the infection. As shown in Fig 4, the CDD model prefers short-term contact at low velocity, while the
proportion of a long contact time is low, indicating that even though the CDD-SEIR model
and the mean-field model obtain qualitatively consistent trends, they are fundamentally differ-
ent. The reason for this lies in the fact that differences in individual motion are considered in
the CDD model so that spatial heterogeneity can be described, while the mean field treatment
ignores such differences. To further illustrate the difference between these two models, we
selected the cases of a relatively low velocity (v = 0.001 m/s, where v < vc) and a medium veloc-
ity case (v = 0.02 m/s, where v > vc) and calculated the total time tSE required for the PS agents
to contact PI agents to become infected. The number of contacts between PS and PI before
infection TSI and the mean and median of contact time per contact between PS and PI before
infection ωSI (please refer to the S1 File for more details) varied with the probability of infec-
tion pSE (Fig 8). At low velocities (v = 0.001 m/s), the PS agent only has 2–3 contacts with PI agents before
becoming infected, and the duration of each contact ωSI tends to be relatively long. This Fig 8. Heterogeneity variable z and spatial heterogeneity This is not the same trend as that obtained by mean-field theory [17, 19]. One possible
reason for the discrepancy obtained by the two models is that the treatment of the mean field
method does not consider spatial heterogeneity, but other factors may also play a role. Fig 7. The slope k (a) and the saturated reproduction number RS (b) influenced by five parameters (nI,0, d, pSE, pEI,Day7, and pIR) in the
system. Fig 7. The slope k (a) and the saturated reproduction number RS (b) influenced by five parameters (nI,0, d, pSE, pEI,Day7, and pIR) in the
system. https://doi.org/10.1371/journal.pone.0286558.g007 PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 11 / 16 PLOS ONE The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading Effect of the CDD model on R0 Contact time required for infection tSE at low velocity (a, v = 0.001 m/s) and medium velocity (b, v = 0.02 m/s), number of contacts
between PS and PI agents before infection TSI, the mean (blue) and median (orange) of the average time per contact between PS and PI
agents before infection ωSI with the probability of infection pSE compared with the contact time ω calculated by the CDD model. At a low
velocity (v = 0.001 m/s), infection tends to occur through a small number of long-term contacts. Fig 8. Contact time required for infection tSE at low velocity (a, v = 0.001 m/s) and medium velocity (b, v = 0.02 m/s), number of contacts
between PS and PI agents before infection TSI, the mean (blue) and median (orange) of the average time per contact between PS and PI
agents before infection ωSI with the probability of infection pSE compared with the contact time ω calculated by the CDD model. At a low
velocity (v = 0.001 m/s), infection tends to occur through a small number of long-term contacts. https://doi.org/10.1371/journal.pone.0286558.g008 PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 12 / 16 PLOS ONE The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading observation indicates that at low velocities, long-term contact is essential for the infection, and
short-term contact is usually not infectious. This can be understood by the longer mean free
time τ at low velocities (Fig 4C). From Eq 3, we can obtain τ 0.22 days when v = 0.001 m/s. In this case, even if all the surroundings are PI agents, it would take several days for PS agents
to accumulate enough short exposures to become infected. However, in the real situation, the
disease state of the infected person cannot be maintained indefinitely, and the whole system
cannot maintain the state of many infected persons for a long time, so the contact time tSE
obtained is often less than the ideal value of 1/pSE (Fig 8A), and there may even be a simulta-
neous decrease in pSE and tSE. Therefore, infections caused by multiple short-term contacts
that take a long time to occur at low velocity are in fact very difficult to occur. This means that
only a small number of prolonged exposures under the CDD model can achieve the conditions
for infection occurrence in the mean field model. Effect of the CDD model on R0 The long-term contact for transmission may
account for strong spatial heterogeneity at low velocity. When the velocity increases (v = 0.02 m/s), it is known by Eq 3 that τ will decrease signifi-
cantly. In this case, the particles do not have to sift through longer collisions used for infection
to occur, and infection can occur through more contacts. Meanwhile, ωSI is also closer to an
ordinary contact (Fig 8B), which is close to the results obtained by the mean-field model. Presumably, allowing short contact exposure with good protective measures can signifi-
cantly reduce R0. Especially when control measures are strong, it is difficult for infections to
reach new susceptible particles, and the effective ability of the virus to spread will be limited. Comparison with other models Various epidemiological models that consider multiple types of compartments beyond the basic
SIR framework have been proposed. In this study, we compare our CDD-SEIR method to two
recently reported methods. One is a random walker’s approach that incorporates the memory
effect into a compartment model [37], where recovered agents are assumed to stay immune for
a random duration. In contrast, our work considers the scenario where the susceptible agents
become carriers after being infected and combines a modified SEIR model with contact-depen-
dent dynamics. Both methods are based on the traditional SIR model and explore the influence
of other factors on space-time dynamics of epidemics. The random walker’s method is efficient
for characterizing epidemics in the long term, while our CDD-SEIR model introduces more
dynamical details by considering patterns of contacts. As the two methods consider different
impacts of other factors on disease transmission, direct comparison of results is challenging. Future studies can further incorporate an immunity impact on transmission in our model. The SEIRS model considers the effects of birth, death, and incubation period on infectious
disease dynamics [38]. Both this model and our work take into account the latency effect,
although in different ways. However, the SEIRS model integrates multiple influences, such as
the length of the latent period, the various routes of transmission, and age-specific transmission. Additionally, the introduction of repeated infection in the SEIRS and random walker’s models
enables the epidemic equilibrium, allowing for longer timescale characterization of the disease
spreading. While our CDD-SEIR model can incorporate these factors by adding agent types to
simulate more realistic scenarios, we primarily focused on the impact of contact due to motion
on epidemic transmission. Although some details of the epidemic transmission process are
excluded, our CDD-SEIR model captures the fundamental information and physical laws. Conclusion In this work, our proposed CDD-SEIR model, which combines the real-scale random motion
of individuals with the traditional compartment model, discloses that epidemic transmission 13 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
June 13, 2023 PLOS ONE The stochastic dynamics disclose spatial heterogeneity about the epidemics spreading depends on spatial heterogeneity and contact patterns. The CDD model has a larger propor-
tion of short-term contacts as the velocity of agents decreases. However, we demonstrate that
the contact pattern of the CDD model with short contact times at low velocity makes the virus
more likely to spread through a small number of long contacts, which causes strong spatial het-
erogeneity and low R0. Due to the presence of spatial heterogeneity, R0 will be proportional to
the logarithm of the group mobility in the strongly heterogeneous range and saturated in the
weakly heterogeneous range, and R0 is approximately independent of the virulence of the virus
when the mobility of the group is low. This demonstrates that in the case of a high reproduc-
tion number, the vaccination policy and reduced contact time can still significantly inhibit the
virus transmission capacity. In the selection of parameters, we tried to make the individual parameters more adapted to
the real scale, which allowed us to obtain a more accurate description of the distribution of
exposure times, thus helping to explain how the movement model and spatial heterogeneity
can influence the spread of epidemics from a microscopic perspective. This is something that
has been relatively lacking in previous work. Such data also fully illustrate the existence of het-
erogeneity in the CDD-SEIR model and the difference with the mean-field model. Overall, we used a real-scale contact-dependent motion model to reveal the effects of spatial
heterogeneity and infection patterns on disease transmission from a microscopic perspective. The insight obtained from this work provides us with a new understanding of how movement
patterns affect the ability of viruses to spread and how to prevent them more effectively. We
hope to improve this CDD-SEIR model in the future to make the movement of individuals
more realistic and thus be able to simulate some of the transmission that is difficult to charac-
terize by numerical or network models. Supporting information S1 File. Computational details and additional data analysis. (DOCX) PLOS ONE | https://doi.org/10.1371/journal.pone.0286558
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June 13, 2023
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A note on secant defective varieties and Clifford modules
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Oliver Nash
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A note on secant defective varieties and Clifford modules
Oliver Nash
Imperial College London
(June 2023)
###### Abstract
We generalise a construction of Landsberg, which associates certain Clifford
algebra representations to Severi varieties. We thus obtain a new proof of
Russo’s Divisibility Property for LQEL varieties.
## 1 Introduction
The geometry of secant-defective varieties is surprisingly rich. In the early
20th Century, the subject captured the attention of several members of the
Italian School of Algebraic Geometry and important results appear in numerous
beautiful old papers, such as those of Scorza [15], Severi [16], and Terracini
[17].
In the 1980s the subject enjoyed a renaissance, largely due to a series of
breakthroughs made by Zak [18]. Zak’s solution of Hartshorne’s linear
normality conjecture [3] lead to his classification of maximally-secant-
deficient varieties, which he named Severi varieties. He showed that there are
exactly four Severi vareties, that they correspond to normed division
algebras, and that their dimensions are 2, 4, 8, 16. The hardest part of the
classification was establishing the dimension restriction.
In 1996, an intruiging paper of Landsberg [8] appeared in which he showed that
the extrinsic geometry of a Severi variety induces certain Clifford algebra
representations. Using the classification of Clifford modules, it is then
trivial to see that the dimensions of the Severi varieties must take the
values already established by Zak.
The main purpose of this note is to show that Landsberg’s Clifford modules may
be generalised. Severi varieties are examples of a class of varieties
introduced by Russo [13] in 2009, called LQEL varieties and we show that
Landsberg’s construction works in this more general setting.
Actually the generalisation is only an extremely mild extension of Landberg’s
results. However we believe it is worth highlighting because, together with
the classification of Clifford modules, it provides a new proof of Russo’s
Divisibility Property for LQEL varieties (corollary 2.8).
## 2 Secant defective varieties and Clifford modules
We shall follow Landsberg [6, 8] closely and so recall his constructions and
notation111Note that [6] (which we follow) adopts slightly different
conventions than [8]. For example $\operatorname{{Ann}}(v)$ in [8] corresponds
to ${\mathbb{P}}\operatorname{{Ann}}(v)$ in [6]..
Our primary object of concern is a subvariety of projective space. We write:
$\displaystyle X\subseteq{\mathbb{P}}^{n+a},$
to indicate that the variety $X$ is $n$-dimensional and that the embedding has
codimension $a$. We work over ${\mathbb{C}}$ throughout, and assume that $X$
is smooth, irreducible, and non-degenerate222Not contained in a hyperplane.,
with secant deficiency $\delta\geq 1$.
### 2.1 Second fundamental form
We recall [2] that the second fundamental form of an embedding
$X\subseteq{\mathbb{P}}^{n+a}$ is a section of
$\operatorname{{Hom}}(S^{2}TX,N)$, where $S^{2}TX$ is the symmetric square of
the tangent bundle and $N$ is the normal bundle. Thus for $x\in X$, we have a
symmetric bilinear map:
$\displaystyle{II}^{x}:S^{2}T_{x}X\to N_{x}.$
When we have chosen a point $x\in X$ and there is no possibility of confusion,
we will write $T$ for $T_{x}X$, $N$ for $N_{x}$ and ${II}$ for ${II}^{x}$.
Taking the transpose, we also regard the second fundamental form as a linear
system of quadrics:
$\displaystyle{II}^{*}:N^{*}\to S^{2}T^{*}.$
A key observation is that global properties of $X$ are visible infinitesimally
via the second fundamental form, and exceptional global properties tend to
produce linear systems of quadrics with exceptional properties.
For example, if $X$ has a smooth dual variety, then at a general point
${II}^{*}$ is a linear system of quadrics of _constant rank_ , and from this
follows Landman’s parity theorem for dual-deficient varieties (see [6, Theorem
12.4.8 and Corollary 12.4.10]).
We shall show that if $X$ has the exceptional property that the Gauss map of a
general tangential projection has zero-dimensional fibres, then its second
fundamental form can be used to construct certain Clifford modules, and from
this follows Russo’s Divisibility Property for secant-deficient varieties (see
[13, Theorem 2.8] or [14, Theorem 4.2.8]).
### 2.2 Tangential projections
We now assume $X$ is non-linear333Note that this is automatic if
$\operatorname{{Sec}}(X)\neq{\mathbb{P}}^{n+a}$ since $X$ is non-degenerate.
make two definitions to fix notation:
###### Definition 2.1.
Let $x\in X$ and let $\mathbb{T}_{x}X\subseteq{\mathbb{P}}^{n+a}$ be the
embedded tangent space at $x$. Away from $\mathbb{T}_{x}X$ we define a
rational map:
$\displaystyle\pi_{x}:X$ $\displaystyle\dashrightarrow{\mathbb{P}}N,$
$\displaystyle y$ $\displaystyle\mapsto[\langle y,\mathbb{T}_{x}X\rangle],$
where $N$ is the fibre of normal bundle of $X$ at $x$. The map $\pi_{x}$ is
known as the tangential projection map at $x$.
We recommend [14, §2.3.2, §3.3] for a valuable discussion of tangential
projections.
###### Definition 2.2.
Let $x\in X$ and ${II}$ be the second fundamental form at $x$. Away from
$\operatorname{{Baseloc}}{II}^{*}$ we define the rational map:
$\displaystyle ii:{\mathbb{P}}T$ $\displaystyle\dashrightarrow{\mathbb{P}}N,$
$\displaystyle[v]$ $\displaystyle\mapsto[{II}(v,v)].$
We recall that the closures of the images of $\pi_{x}$ and $ii$ coincide and
have dimension $n-\delta$ (see e.g., [14, Proposition 2.3.5] and its proof).
Let this $(n-\delta)$-dimensional irreducible subvariety be:
$\displaystyle Z\subseteq{\mathbb{P}}N,$
and let:
$\displaystyle\operatorname{{Sec}}(X)\subseteq{\mathbb{P}}^{n+a},$
be the $(2n+1-\delta)$-dimensional secant variety of $X$, then we note for
future reference that:
$\displaystyle\operatorname{{codim}}Z=\operatorname{{codim}}\operatorname{{Sec}}(X).$
(1)
### 2.3 Second fundamental form of a tangential projection
The following is essentially a restatement of [8, Lemma 6.6].
Given a general point $x\in X$ and a general444Thus $v$ is non-zero and
$[v]\not\in\operatorname{{Baseloc}}{II}^{*}$. vector $v\in T$, let
$z=ii([v])$. It follows from the definition of $ii$ that there is a natural
commutative diagram:
where $ii_{*[v]}$ is the derivative of $ii$ at $[v]$ and ${II}_{v}$ is the
map:
$\displaystyle{II}_{v}:w\mapsto{II}(v,w).$
We thus have natural exact sequences:
$\displaystyle 0\to\langle v,\ker{II}_{v}\rangle\to
T\xrightarrow{\rho_{T}}T_{z}Z\to 0,$ (2)
and:
$\displaystyle 0\to{II}_{v}(T)\to N\xrightarrow{\rho_{N}}N_{z}Z\to 0,$
where $N_{z}Z$ is the normal bundle of $Z\subseteq{\mathbb{P}}N$ at $z$. The
maps $\rho_{T}$ and $\rho_{N}$ fit into the following commutative diagram:
$\displaystyle\begin{gathered}\lx@xy@svg{\hbox{\raise 0.0pt\hbox{\kern
24.46875pt\hbox{\ignorespaces\ignorespaces\ignorespaces\hbox{\vtop{\kern
0.0pt\offinterlineskip\halign{\entry@#!@&&\entry@@#!@\cr&\\\&\crcr}}}\ignorespaces{\hbox{\kern-11.37048pt\raise
0.0pt\hbox{\hbox{\kern 0.0pt\raise 0.0pt\hbox{\hbox{\kern 3.0pt\raise
0.0pt\hbox{$\textstyle{S^{2}T\ignorespaces\ignorespaces\ignorespaces\ignorespaces\ignorespaces\ignorespaces\ignorespaces\ignorespaces}$}}}}}}}\ignorespaces\ignorespaces\ignorespaces\ignorespaces{}{\hbox{\lx@xy@droprule}}\ignorespaces\ignorespaces\ignorespaces{\hbox{\kern
20.03621pt\raise 5.39166pt\hbox{{}\hbox{\kern 0.0pt\raise
0.0pt\hbox{\hbox{\kern 3.0pt\hbox{\hbox{\kern
0.0pt\raise-2.39166pt\hbox{$\scriptstyle{{II}}$}}}\kern
3.0pt}}}}}}\ignorespaces{\hbox{\kern 45.7627pt\raise 0.0pt\hbox{\hbox{\kern
0.0pt\raise
0.0pt\hbox{\lx@xy@tip{1}\lx@xy@tip{-1}}}}}}{\hbox{\lx@xy@droprule}}{\hbox{\lx@xy@droprule}}\ignorespaces\ignorespaces\ignorespaces\ignorespaces{}{\hbox{\lx@xy@droprule}}\ignorespaces\ignorespaces\ignorespaces{\hbox{\kern-24.46875pt\raise-19.62025pt\hbox{{}\hbox{\kern
0.0pt\raise 0.0pt\hbox{\hbox{\kern 3.0pt\hbox{\hbox{\kern
0.0pt\raise-1.35834pt\hbox{$\scriptstyle{\rho_{T}\otimes\rho_{T}}$}}}\kern
3.0pt}}}}}}\ignorespaces{\hbox{\kern 0.0pt\raise-27.60277pt\hbox{\hbox{\kern
0.0pt\raise
0.0pt\hbox{\lx@xy@tip{1}\lx@xy@tip{-1}}}}}}{\hbox{\lx@xy@droprule}}{\hbox{\lx@xy@droprule}}{\hbox{\kern
45.7627pt\raise 0.0pt\hbox{\hbox{\kern 0.0pt\raise 0.0pt\hbox{\hbox{\kern
3.0pt\raise
0.0pt\hbox{$\textstyle{N\ignorespaces\ignorespaces\ignorespaces\ignorespaces}$}}}}}}}\ignorespaces\ignorespaces\ignorespaces\ignorespaces{}{\hbox{\lx@xy@droprule}}\ignorespaces\ignorespaces\ignorespaces{\hbox{\kern
53.32518pt\raise-19.62025pt\hbox{{}\hbox{\kern 0.0pt\raise
0.0pt\hbox{\hbox{\kern 3.0pt\hbox{\hbox{\kern
0.0pt\raise-0.82361pt\hbox{$\scriptstyle{\rho_{N}}$}}}\kern
3.0pt}}}}}}\ignorespaces{\hbox{\kern
53.32518pt\raise-29.40721pt\hbox{\hbox{\kern 0.0pt\raise
0.0pt\hbox{\lx@xy@tip{1}\lx@xy@tip{-1}}}}}}{\hbox{\lx@xy@droprule}}{\hbox{\lx@xy@droprule}}{\hbox{\kern-16.56659pt\raise-39.24052pt\hbox{\hbox{\kern
0.0pt\raise 0.0pt\hbox{\hbox{\kern 3.0pt\raise
0.0pt\hbox{$\textstyle{S^{2}T_{z}Z\ignorespaces\ignorespaces\ignorespaces\ignorespaces}$}}}}}}}\ignorespaces\ignorespaces\ignorespaces\ignorespaces{}{\hbox{\lx@xy@droprule}}\ignorespaces\ignorespaces\ignorespaces{\hbox{\kern
20.8848pt\raise-32.62941pt\hbox{{}\hbox{\kern 0.0pt\raise
0.0pt\hbox{\hbox{\kern 3.0pt\hbox{\hbox{\kern
0.0pt\raise-3.61111pt\hbox{$\scriptstyle{\widetilde{II}}$}}}\kern
3.0pt}}}}}}\ignorespaces{\hbox{\kern
40.56659pt\raise-39.24052pt\hbox{\hbox{\kern 0.0pt\raise
0.0pt\hbox{\lx@xy@tip{1}\lx@xy@tip{-1}}}}}}{\hbox{\lx@xy@droprule}}{\hbox{\lx@xy@droprule}}{\hbox{\kern
40.56659pt\raise-39.24052pt\hbox{\hbox{\kern 0.0pt\raise
0.0pt\hbox{\hbox{\kern 3.0pt\raise
0.0pt\hbox{$\textstyle{N_{z}Z}$}}}}}}}\ignorespaces}}}}\ignorespaces\end{gathered}$
(8)
where $\widetilde{II}$ is the second fundamental form of $Z$ at $z$.
### 2.4 Singular locus of the second fundamental form
Griffiths and Harris noticed that the quadrics of the second fundamental form
are all singular along the fibres of the Gauss map. In fact they proved [2,
(2.6)]:
$\displaystyle\operatorname{{Singloc}}(N^{*})=T_{x}F,$
where $F$ is the fibre of the Gauss map through $x$ and for any $A\subseteq
N^{*}$, $\operatorname{{Singloc}}(A)$ is the intersection of all the singular
loci:
$\displaystyle\operatorname{{Singloc}}(A)=\bigcap_{f\in A}\\{v\in
T~{}|~{}v\righthalfcup{II}^{*}(f)=0\\}.$ (9)
A key insight of Landsberg [8] was that there are natural subsystems
$A\subseteq N^{*}$ for which $\operatorname{{Singloc}}(A)$ captures more
delicate geometric features of $X$. Indeed it follows from (8) that there is a
natural exact sequence:
$\displaystyle
0\to\ker\rho_{T}\to\operatorname{{Singloc}}(\rho_{N}^{*}(N^{*}_{z}Z))\to\operatorname{{Singloc}}(N_{z}^{*}Z)\to
0,$ (10)
and so for $v\in T$, Landsberg defined:
$\displaystyle\operatorname{{Ann}}(v)$
$\displaystyle=\rho_{N}^{*}(N^{*}_{z}Z)$ $\displaystyle={II}_{v}(T)^{\perp}$
$\displaystyle=\\{f\in N^{*}~{}|~{}v\righthalfcup{II}^{*}(f)=0\\},$
and studied the middle term
$\operatorname{{Singloc}}(\operatorname{{Ann}}(v))$ appearing in (10).
We note for future reference that:
$\displaystyle\dim\operatorname{{Ann}}(v)$
$\displaystyle=\operatorname{{codim}}Z$
$\displaystyle=\operatorname{{codim}}\operatorname{{Sec}}(X)\quad\mbox{by
\eqref{eq:codim_sec_codim_Z}}.$ (11)
### 2.5 Clifford modules
The simplest class of secant-deficient varieties are those for which the Gauss
map of a general tangential projection has zero-dimensional fibres. For
emphasis we state a key consequence of this property:
###### Lemma 2.3.
Let $X\subseteq{\mathbb{P}}^{n+a}$ be a smooth, irreducible, non-degenerate
variety of secant deficiency $\delta\geq 1$. Let $x\in X$, $v\in T_{x}X$ be
general and let $Z\subseteq{\mathbb{P}}N$ be the closure of the tangential
projection at $x$. Then the Gauss map of $Z$ has zero-dimensional fibres if
and only if:
$\displaystyle\langle
v,\ker{II}_{v}\rangle=\operatorname{{Singloc}}(\operatorname{{Ann}}(v)).$ (12)
###### Proof.
Applying Griffiths and Harris’s result [2, (2.6)] to $Z$, we know that the
Gauss map of $Z$ has zero-dimensional fibres if and only if the third term in
(10) vanishes. Bearing in mind (2), the conclusion is clear. ∎
###### Remark 2.4.
The Scorza Lemma [14, Theorem 3.3.3] tells us that the varieties satisfying
the conditions of lemma 2.3 are LQEL varieties. Moreover we have examples:
* •
the quadratic Veronese embeddings
$\nu_{2}({\mathbb{P}}^{n})\subseteq{\mathbb{P}}^{n(n+3)/2}$ for $n\geq 2$
($\delta=1$),
* •
the binary Segre embeddings
${\mathbb{P}}^{n}\times{\mathbb{P}}^{m}\subseteq{\mathbb{P}}^{mn+m+n}$ for
$m+n\geq 3$ ($\delta=2$),
* •
the rank-2 Plücker embeddings $G(2,n)\subseteq{\mathbb{P}}^{(n-2)(n+1)/2}$ for
$n\geq 5$ ($\delta=4$),
* •
the 16-dimensional Severi variety in ${\mathbb{P}}^{26}$ ($\delta=8$),
as well as their linear projections. We recommend [14] for further discussion.
###### Remark 2.5.
In [14, Definition 3.3.1], given general points $x,y\in X$ and general
$p\in\operatorname{{Sec}}(X)$ on the line $\overline{xy}$, Russo defines the
contact locus $\Gamma_{p}\subseteq X$ as:
$\displaystyle\Gamma_{p}=\overline{\\{x\in X~{}|~{}T_{x}X\subseteq
T_{p}\operatorname{{Sec}}(X)\\}},$
and notes that by Terracini’s lemma:
$\displaystyle\Sigma_{p}\subseteq\Gamma_{p},$
where $\Sigma_{p}$ is the entry locus of $X$ with respect to
$p\in\operatorname{{Sec}}(X)$.
Let $Z=\overline{\pi_{x}(X)}$ be the closure of the tangential projection at
$x$ and $F\subseteq Z$ be the fibre of the Gauss map of $Z$ through
$z=\pi_{x}(y)$. As argued by Russo in the proof of [14, Lemma 3.3.2] the
irreducible components of $\pi_{x}^{-1}(F)$ and $\Gamma_{p}$ through $y$
coincide. We should thus have a natural exact sequence of tangent spaces:
$\displaystyle 0\to T_{y}\Sigma_{p}\to T_{y}\Gamma_{p}\to T_{z}F\to 0.$ (13)
The line $\overline{yp}$ naturally determines a vector $v\in T_{y}X$, and we
expect:
$\displaystyle\operatorname{{Singloc}}(Ann(v))=T_{y}\Gamma_{p},$
so that (13) can be interpreted as (10). Given this, [14, Lemma 3.3.2 (2)]
would provide an alternate proof of lemma 2.3 above.
We come at last to our main point:
###### Theorem 2.6.
Let $X\subseteq{\mathbb{P}}^{n+a}$ be an smooth, irreducible, non-degenerate
variety of secant deficiency $\delta\geq 1$ such that
$\operatorname{{Sec}}(X)\neq{\mathbb{P}}^{n+a}$. Suppose that the Gauss map of
the tangential projection at a general point $x$ has zero-dimensional fibres,
and let $v\in T_{x}X$ be general. Then
$T/\operatorname{{Singloc}}(\operatorname{{Ann}}(v))$ carries a natural
Clifford module structure over $\ker{II}_{v}$.
###### Proof.
Let $Z=\overline{\pi_{x}(X)}\subseteq{\mathbb{P}}N$ be the closure of the
image of the tangential projection at $x$.
The result we need is exactly [8, Lemma 6.26] except that we have made no
assumption about $Z$ being a cone (instead assuming that its Gauss map has
zero-dimensional fibres) and we do not assume that $Z$ is a hypersurface. In
view of (1), $Z$ is a hypersurface if and only if $\operatorname{{Sec}}(X)$
is. However since the linear projection from a linear subspace which does not
meet $\operatorname{{Sec}}(X)$ is an isomorphism, we may select a maximal such
subspace and project down to the case that $\operatorname{{Sec}}(X)$ is a
hypersurface; the lemma then applies, and our proof is complete.
For the benefit of readers who wish to compare with [6], we provide a
reference for the argument as presented there. The key equation is [6, (12.22)
page 374]:
$\displaystyle q^{n+j}_{\epsilon\kappa}q^{n+k}_{\delta i}+q^{n+j}_{\delta
k}q^{n+k}_{\epsilon
i}=-2q^{n+1}_{\epsilon\delta}\delta^{i}_{j}\quad\forall\epsilon,\delta,j,k,i.$
The key assumption required for the derivation is $S=0$ where:
$\displaystyle
S=\dim\operatorname{{Singloc}}\operatorname{{Ann}}(v)-\dim\langle
v,\ker{II}_{v}\rangle,$
which follows from lemma 2.3. ∎
###### Remark 2.7.
We can restate theorem 2.6 without referring to the second fundamental form as
follows.
Let $X$ be as in theorem 2.6 and let $p\in\operatorname{{Sec}}(X)$ and $x\in
X$ be general points. Let $Q\subseteq X$ be the irreducible component of the
$p$-entry locus through $x$ and let $Q^{\prime}\subseteq Q$ be the
corresponding555See [11, Lemma 2.4]. irreducible component of the tangent
locus through $x$. Then $T_{x}Q^{\prime}$ carries a non-degenerate quadratic
form and the fibre $N^{x}_{Q|X}$ of the normal bundle of $Q$ in $X$ is a
Clifford module for the Clifford algebra $Cl(T_{x}Q^{\prime})$.
We emphasise the following corollary:
###### Corollary 2.8.
Let $X$ be as in theorem 2.6 then:
$\displaystyle\left.2^{\left\lfloor\frac{\delta-1}{2}\right\rfloor}~{}\right\mid~{}n-\delta.$
###### Proof.
The result follows immediately from theorem 2.6 together with the
classification of Clifford modules. Indeed if there exists a $k$-dimensional
Clifford module of a non-degenerate $l$-dimensional complex quadratic form,
then:
$\displaystyle\left.p~{}\right\mid~{}k,$
where $p=2^{\left\lfloor\frac{l}{2}\right\rfloor}$. The reason is that the
Clifford algebra of the quadratic form is the matrix algebra
${\mathbb{C}}^{p\times p}$ if $l$ is even or ${\mathbb{C}}^{p\times
p}\oplus{\mathbb{C}}^{p\times p}$ if $l$ is odd (see e.g., [1, Table 1]) and
the natural action of ${\mathbb{C}}^{p\times p}$ on ${\mathbb{C}}^{p}$ is its
unique irreducible representation. ∎
###### Remark 2.9.
The divisibility condition established in corollary 2.8 was first proved by
Russo and appeared in [13, Theorem 2.8] (see also [14, Theorem 4.2.8]). The
proof involved an inductive study of the Hilbert scheme of lines through a
general point of an LQEL variety.
A second proof (due to the author) appeared as [11, Corollary 2.6]. This proof
was topological and the key was a calculation in topological $K$-theory.
We now have a third proof (albeit with slightly different assumptions) and
this time it is Clifford module theory that is the key.
It would be interesting to explore the relationship between the second and
third proofs given the deep connections between $K$-theory and Clifford
modules identified by Atiyah, Bott, and Shapiro in [1]. The first step should
be to argue that Landsberg’s construction actually provides bundles of
representations of Clifford algebras, as $x$ varies over a general tangent
locus.
###### Remark 2.10.
Note that the proof of corollary 2.8 shows that the 2 which appears in the
expression $(\delta-1)/2$ corresponds to the mod-2 periodicity of Morita
equivalence classes of complex Clifford algebras. Thus it is the same 2 which
appears in complex Bott periodicity.
###### Remark 2.11.
A quite different connection between Clifford modules and Severi varieties
arises in the context of ‘Clifford structures’, introduced by Moroianu and
Semmelmann in [10]. The Severi varieties appear in the classification of
parallel non-flat even Clifford structures in [10] (see also [12]). It might
be interesting to explore whether these Clifford structures have any
relationship to Landsberg’s Clifford modules.
## 3 A remark about the $\delta\leq 8$ problem
Let $X\subseteq{\mathbb{P}}^{n+a}$ be a smooth, irreducible, non-degenerate,
subvariety with $\operatorname{{Sec}}(X)\neq{\mathbb{P}}^{n+a}$. It follows
from Zak’s proof of Hartshorne’s linear normality conjecture that the secant
deficiency satisfies:
$\displaystyle\delta\leq\left\lfloor\frac{n}{2}\right\rfloor.$ (14)
In the course of their exposition [9] of Zak’s work, Lazarsfeld and Van de Ven
highlighted that all known examples of $X$ as above satisfy $\delta\leq 8$.
They thus posed the problem to investigate whether $\delta$ could be
arbitrarily large (see [9, §1f, page 19]). In view of (14), any $X$ with
$\delta>8$ must have dimension $n\geq 18$.
Very little progress has been made on this problem in the 40 years since it
was first posed. Kaji [7] has shown that any variety with $\delta>8$ must be
non-homogeneous but otherwise the problem remains completely open: all known
examples still satisfy $\delta\leq 8$ and the 16-dimensional Severi variety
remains the only variety known to achieve $\delta=8$. The problem remains open
even for the very special class of LQEL varieties (see [14, chapter 4.4, page
113] as well as [4, Remark 3.8] and [5, Conjecture 4.5]).
We mention this problem here, to highlight that by combining known results,
one may sharpen (14) slightly as follows:
###### Lemma 3.1.
Let $X\subseteq{\mathbb{P}}^{n+a}$ be a smooth, irreducible, non-degenerate
subvariety with $\operatorname{{Sec}}(X)\neq{\mathbb{P}}^{n+a}$. Suppose
$n\geq 17$, then:
$\displaystyle\delta\leq\left\lfloor\frac{n-1}{2}\right\rfloor.$
###### Proof.
For a general tangential projection of $X$, let $\tilde{\gamma}$ be the
dimension of the fibre of its Gauss map and $\tilde{\xi}$ its dual deficiency.
Suppose first that $\tilde{\gamma}=0$. We may assume $\delta\geq 1$ (otherwise
there is nothing to prove) so by Scorza’s Lemma [14, Theorem 3.3.3] $X$ is an
LQEL variety. By [14, Corollary 4.4.11]:
$\displaystyle\delta\leq\left\lfloor\frac{n+8}{3}\right\rfloor\leq\left\lfloor\frac{n-1}{2}\right\rfloor$
since $n\geq 17$.
It remains to consider the case $\tilde{\gamma}\geq 1$. By [14, Theorem 5.4.1,
Lemma 3.3.2]:
$\displaystyle\delta\leq\frac{n-\tilde{\xi}}{2}\leq\frac{n-\tilde{\gamma}}{2}\leq\frac{n-1}{2},$
as required666We note in passing that one could instead deal with the case
$\tilde{\gamma}\geq 1$ using results of Landsberg. Indeed after projecting to
ensure $\operatorname{{Sec}}(X)$ is a hypersurface, one could apply [8,
Corollary 7.3] (equivalently, the inequality at the bottom of page 373 of
[6]).. ∎
Note that lemma 3.1 increases the dimension restriction on a variety with
$\delta>8$ slightly to $n\geq 19$. One might hope to make further progress by
studying varieties for which $\tilde{\gamma}=1$ and then arguing as in lemma
3.1 but with three cases corresponding to whether $\tilde{\gamma}$ is 0, 1, or
at least 2.
## References
* [1] M. F. Atiyah, R. Bott, and A. Shapiro. Clifford modules. Topology, 3(suppl. 1):3–38, 1964.
* [2] Phillip Griffiths and Joseph Harris. Algebraic geometry and local differential geometry. Ann. Sci. École Norm. Sup. (4), 12(3):355–452, 1979.
* [3] Robin Hartshorne. Varieties of small codimension in projective space. Bull. Amer. Math. Soc., 80:1017–1032, 1974.
* [4] Paltin Ionescu and Francesco Russo. Varieties with quadratic entry locus. II. Compos. Math., 144(4):949–962, 2008.
* [5] Paltin Ionescu and Francesco Russo. On dual defective manifolds. Math. Res. Lett., 21(5):1137–1154, 2014.
* [6] Thomas A. Ivey and Joseph M. Landsberg. Cartan for beginners, volume 175 of Graduate Studies in Mathematics. American Mathematical Society, Providence, RI, 2016. Differential geometry via moving frames and exterior differential systems, Second edition [of MR2003610].
* [7] Hajime Kaji. Homogeneous projective varieties with degenerate secants. Trans. Amer. Math. Soc., 351(2):533–545, 1999.
* [8] J. M. Landsberg. On degenerate secant and tangential varieties and local differential geometry. Duke Math. J., 85(3):605–634, 1996.
* [9] R. Lazarsfeld and A. Van de Ven. Topics in the geometry of projective space, volume 4 of DMV Seminar. Birkhäuser Verlag, Basel, 1984. Recent work of F. L. Zak, With an addendum by Zak.
* [10] Andrei Moroianu and Uwe Semmelmann. Clifford structure on Riemannian manifolds. Adv. Math., 228(2):940–967, 2011.
* [11] Oliver Nash. $K$-theory, LQEL manifolds and Severi varieties. Geom. Topol., 18(3):1245–1260, 2014.
* [12] Maurizio Parton and Paolo Piccinni. The even Clifford structure of the fourth Severi variety. Complex Manifolds, 2(1):89–104, 2015.
* [13] Francesco Russo. Varieties with quadratic entry locus. I. Math. Ann., 344(3):597–617, 2009.
* [14] Francesco Russo. On the geometry of some special projective varieties, volume 18 of Lecture Notes of the Unione Matematica Italiana. Springer, Cham; Unione Matematica Italiana, Bologna, 2016.
* [15] Gaetano Scorza. Sulle varietá a quattro dimensioni di sr ($r\geq 9$) i cui $s_{4}$ tangenti si tagliano a due a due. Rend. Circ. Mat. Palermo, 27:148–178, 1909.
* [16] Francesco Severi. Intorno ai punti doppi impropri di una superficie generale dello spazio a quattro dimensioni, e a’ suoi punti tripli apparenti. Rendiconti del Circolo Matematico di Palermo, 15(1):33–51, 1901\.
* [17] Alessandro Terracini. Sulle $v_{k}$ per cui la varietà degli $s_{h}(h+1)$-seganti ha dimensione minore dell’ordinario. Rend. Circ. Mat. Palermo, 31:392–396, 1911.
* [18] F. L. Zak. Tangents and secants of algebraic varieties, volume 127 of Translations of Mathematical Monographs. American Mathematical Society, Providence, RI, 1993. Translated from the Russian manuscript by the author.
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Low medication adherence in patients with Short Bowel Syndrome dependent on parenteral nutrition
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Iahel M. L. Ferreira
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ARTIGO ORIGINAL ARTIGO ORIGINAL RESUMO Modelo do estudo: Transversal. Objetivo do estudo: Avaliar a adesão de pacientes com Síndrome do
Intestino Curto ao tratamento medicamentoso por via oral. Casuística e Métodos: O estudo inclui 10
pacientes com Síndrome do Intestino Curto (SIC) acompanhados em unidade especializada de um
hospital universitário. Todos os pacientes recebiam nutrição parenteral por no mínimo quatro meses. A
casuística consistia em seis homens e quatro mulheres, com idade de 56 ± 13 anos e Índice de Massa
Corporal de 18 ± 3kg/m2. Foram registradas informações sobre o uso de medicamentos no domicílio. A
adesão ao tratamento medicamentoso foi avaliada pelo teste de Morisky e foi identificada a natureza do
comportamento em relação à não adesão. Resultados Apenas 40% dos pacientes foram classificados
como “mais aderentes”. Dentre os “menos aderentes”, a baixa adesão por associação do comportamen-
to intencional e não intencional foi observada em três casos; dois pacientes apresentavam apenas o
comportamento intencional e o comportamento não intencional foi documentado em um paciente. Con-
clusões: A maioria dos pacientes tem baixa adesão ao tratamento medicamentoso por via oral durante
o período em que eles permanecem em seus domicílios. Tais resultados sugerem o desenvolvimento e a
implementação de estratégias que visem à percepção da relevância do uso correto dos medicamentos e
a promoção da adesão ao tratamento medicamentoso. Palavras-chave: Adesão à Medicação. Síndrome do Intestino Curto. Tratamento. Nutrição Pare Low medication adherence in patients with Short Bowel Syndrome dependent on
parenteral nutrition Iahel M.L. Ferreira1, Camila B.M. Braga2, Nathalie L.S. Dewulf3, Julio S. Marchini4, Selma F.C. Cunha4 Baixa adesão ao uso de medicamentos em pacientes
com Síndrome do Intestino Curto dependentes de
nutrição parenteral Low medication adherence in patients with Short Bowel Syndrome dependent on
parenteral nutrition Medicina (Ribeirão Preto. Online) 2016;49(5):429-434 1. Farmacêutica, mestre. Divisão de Nutrologia, Departamen-
to de Clínica Médica, Faculdade de Medicina de Ribeirão
Preto da Universidade de São Paulo (FMRP–USP). Financiamento: Fundação de Pesquisa do Estado de São Paulo (#)
Conflitos de interesse: Os autores declaram ausência de conflito de i ABSTRACT Information related to the use
of medications at home was recorded, and adherence behavior was assessed by using the Morisky
Medication Adherence Scale. Results: Four patients (40%) had high medication adherence. Among
patients with low medication adherence, unintentional behavior was observed in two patients, inten-
tional behavior was observed in one patient, and an association of unintentional with intentional behavior
was identified in three patients. Conclusion: Outside the hospital setting, most patients with SBS have
low adherence to oral medications. These results suggest the development and implementation of tar-
geted strategies aimed to promote awareness and adherence to medication treatment. Key-word: Medication Adherence. Short Bowel Syndrome. Therapeutics. Parenteral Nutrition. ABSTRACT Study design: Cross-sectional. Aim: We aimed to investigate adherence to oral medications in pa-
tients with severe Short Bowel Syndrome (SBS). Casuistic and Methods: Ten patients with severe
intestinal failure attending a specialized unit in a university hospital were enrolled in this study. All Correspondência
Selma Freire de Carvalho da Cunha
Hospital das Clínicas da FMRP–USP
Departamento de Clínica Médica, 6º. Andar
Avenida dos Bandeirantes, 3900
CEP 14048-90 - Ribeirão Preto – SP – Brasil Correspondência
Selma Freire de Carvalho da Cunha
Hospital das Clínicas da FMRP–USP
Departamento de Clínica Médica, 6º. Andar
Avenida dos Bandeirantes, 3900
CEP 14048-90 - Ribeirão Preto – SP – Brasil 3. Farmacêutica, doutora. Faculdade de Farmácia. Universi-
dade Federal de Goiás. 4. Docentes. Médicos, doutores. Divisão de Nutrologia, FMRP–
USP. 4. Docentes. Médicos, doutores. Divisão de Nutrologia, FMRP–
USP. Recebido em 25/05/2015
Aprovado em 09/03/2016 Financiamento: Fundação de Pesquisa do Estado de São Paulo (#)
Conflitos de interesse: Os autores declaram ausência de conflito de interesse Financiamento: Fundação de Pesquisa do Estado de São Paulo (#)
Conflitos de interesse: Os autores declaram ausência de conflito de interesse Medicina (Ribeirão Preto. Online) 2016;49(5):429-434 DOI: http://dx.doi.org/10.11606/issn.2176-7262.v49i5p429-434 Ferreira IML, Braga CBM, Dewulf NLS, Marchini JS, Cunha SFC. Baixa adesão à medicamentos na Síndrome do Intestino Curto. Medicina (Ribeirão Preto. Online) 2016;49(5):429-34 Ferreira IML, Braga CBM, Dewulf NLS, Marchini JS, Cunha SFC. Baixa adesão à medicamentos na Síndrome do Intestino Curto. patients received intermittent parenteral nutrition for at least 4 months. The participants (six men and
four women) had 56 ± 13 years and body mass index of 18 ± 3 kg/m2. Information related to the use
of medications at home was recorded, and adherence behavior was assessed by using the Morisky
Medication Adherence Scale. Results: Four patients (40%) had high medication adherence. Among
patients with low medication adherence, unintentional behavior was observed in two patients, inten-
tional behavior was observed in one patient, and an association of unintentional with intentional behavior
was identified in three patients. Conclusion: Outside the hospital setting, most patients with SBS have
low adherence to oral medications. These results suggest the development and implementation of tar-
geted strategies aimed to promote awareness and adherence to medication treatment. patients received intermittent parenteral nutrition for at least 4 months. The participants (six men and
four women) had 56 ± 13 years and body mass index of 18 ± 3 kg/m2. Introdução mais confiáveis, mas apresentam custo mais eleva-
do, são mais invasivos e de aplicação restrita.17 O
conhecimento demonstrado pelo paciente sobre os
medicamentos utilizados e suas respectivas
posologias pode ser um indício indireto de sua ade-
são.18 Embora possam superestimar a adesão ao
tratamento, os métodos indiretos são largamente
empregados em pesquisas e incluem entrevistas
estruturadas, com a vantagem de serem de fácil
acesso e de custo reduzido. A Síndrome do Intestino Curto (SIC) resulta
em subnutrição, diarreia, desidratação e distúrbios
eletrolíticos secundários à má-absorção por redu-
ção na área intestinal funcionante.1,2 Pacientes com
SIC necessitam de longos períodos de hospitaliza-
ções3 e a nutrição parenteral e a reposição endove-
nosa hidroeletrolítica e de vitaminas são essenciais
após ressecções intestinais extensas.4,5,6 O quadro
disabsortivo pode manter-se indefinidamente de-
pendendo da extensão da ressecção intestinal e da
capacidade absortiva do intestino remanescente. A
abordagem terapêutica em tais casos visa capaci-
tar o paciente ao retorno às suas atividades habitu-
ais, que implica no controle da diarreia, na manu-
tenção do peso corporal aceitável e na redução ou
mesmo a eliminação da necessidade da nutrição pa-
renteral.7,8 Após a ressecção do intestino delgado,
as principais mudanças comportamentais visam a
adesão às orientações dietéticas, ao uso de medi-
cação e à suplementação vitamínica e mineral.9,10 A
evolução favorável depende da adesão ao tratamen-
to dietético e farmacológico, que têm o potencial
de melhorar as condições clínicas e nutricionais,
além da qualidade de vida dos pacientes.11,12,13 Nós não temos conhecimento de estudos que
avaliem a adesão ao tratamento medicamentoso oral
em pacientes com SIC. Nesse contexto, o objetivo
do presente estudo foi avaliar a adesão de pacien-
tes com SIC ao tratamento medicamentoso por via
oral. http://www.revistas.usp.br/rmrp / http://revista.fmrp.usp.br Ferreira IML, Braga CBM, Dewulf NLS, Marchini JS, Cunha SFC.
Baixa adesão à medicamentos na Síndrome do Intestino Curto. Avaliação da adesão ao tratamento Os voluntários da pesquisa foram questiona-
dos quanto à terapia medicamentosa domiciliar. A
adesão ao tratamento medicamentoso foi avaliada
utilizando-se o Teste de Moriksy (1986)15, aplicado
durante a hospitalização, período em que os pa-
cientes recebiam nutrição parenteral. O teste cons-
titui-se de questionário validado, estruturado, com
quatro perguntas fechadas e respostas dicotômicas. De acordo com o número de respostas “sim”, a ade-
são foi classificada em dois níveis: “menos aderen-
te” (1 a 4 respostas “sim”) e “mais aderente” (ne-
nhuma resposta “sim”). A não adesão ao tratamen-
to medicamentoso foi classificada como intencional
ou não intencional, de acordo com os comporta-
mentos mostrados no Quadro 1.19 A caracterização da casuística foi feita a par-
tir de informações obtidas no prontuário médico dos
pacientes. O gênero dos voluntários, a idade e o
índice de massa corporal (IMC, kg/m2) no momen-
to da avaliação foram registrados, assim como a
etiologia da SIC, o tempo de ressecção intestinal, a
extensão do intestino delgado remanescente e as
doenças concomitantes (Tabela 1). Foram registra- Casuística opção disponível no sistema de saúde público bra-
sileiro, os pacientes com falência intestinal grave
recebem nutrição parenteral intermitente de acordo
com a rotina do nosso serviço.4,5,6 Durante cada ad-
missão hospitalar, os pacientes recebem a nutrição
parenteral por cinco a oito dias, com intervalo de 10
a 40 dias entre as hospitalizações, dependendo da
intensidade da diarreia, da ocorrência de desidrata-
ção e da piora do estado clínico ou nutricional. To-
dos os pacientes recebem aconselhamento do nutri-
cionista para seguimento da dieta por via oral no
domicilio. Ainda, a equipe medica prescreve suple-
mentos de vitaminas e minerais para uso diário como
o Centrum® (Wyeth, Itapevi, Brasil) e Materna®
(Wyeth, Itapevi, Brasil), solução de reidratação oral,
antidiarreicos e medicamentos para controle das
comorbidades apresentadas. A cada internação os
pacientes recebem informações sobre sua doença e
a importância da adesão ao tratamento. das as informações sobre o uso de medicamentos
no intervalo entre internações em que eles recebi-
am nutrição parenteral. Casuística O estudo foi conduzido em uma unidade
especializada para tratamento de pacientes com
SIC de um hospital universitário brasileiro, após
aprovação pelo Comitê de Ética em Pesquisa da Ins-
tituição (Processo no. 11442/07). A coleta dos da-
dos foi feita no período de agosto de 2008 a no-
vembro de 2009 e o estudo incluiu todos os pacien-
tes com falência intestinal grave que estavam em
acompanhamento na unidade por período mínimo
de quatro meses, a fim de garantir que os voluntá-
rios tivessem conhecimento sobre a rotina do tra-
tamento. A baixa adesão à terapêutica representa um
dos maiores problemas clínicos no controle de con-
dições que requerem tratamento farmacológico e
mudanças no estilo de vida.14,15,16 A avaliação da
adesão ao tratamento permite classificar o compor-
tamento do paciente em relação às recomendações
do profissional da área da saúde, seja sobre o uso
de medicamentos, dietas ou mudanças no estilo de
vida.17 Os métodos diretos de avaliação da adesão
ao tratamento medicamentoso, como as dosagens
séricas ou urinárias de determinado fármaco, são A casuística foi composta de seis homens e
quatro mulheres com idade média de 56 ± 13 anos
e Índice de Massa Corporal (IMC) de 18 ± 3kg/m2. Considerando que a nutrição parenteral não é uma 430 http://www.revistas.usp.br/rmrp / http://revista.fmrp.usp.br Medicina (Ribeirão Preto. Online) 2016;49(5):429-34 Ferreira IML, Braga CBM, Dewulf NLS, Marchini JS, Cunha SFC. Baixa adesão à medicamentos na Síndrome do Intestino Curto. Ferreira IML, Braga CBM, Dewulf NLS, Marchini JS, Cunha SFC. Baixa adesão à medicamentos na Síndrome do Intestino Curto. opção disponível no sistema de saúde público bra-
sileiro, os pacientes com falência intestinal grave
recebem nutrição parenteral intermitente de acordo
com a rotina do nosso serviço.4,5,6 Durante cada ad-
missão hospitalar, os pacientes recebem a nutrição
parenteral por cinco a oito dias, com intervalo de 10
a 40 dias entre as hospitalizações, dependendo da
intensidade da diarreia, da ocorrência de desidrata-
ção e da piora do estado clínico ou nutricional. To-
dos os pacientes recebem aconselhamento do nutri-
cionista para seguimento da dieta por via oral no
domicilio. Ainda, a equipe medica prescreve suple-
mentos de vitaminas e minerais para uso diário como
o Centrum® (Wyeth, Itapevi, Brasil) e Materna®
(Wyeth, Itapevi, Brasil), solução de reidratação oral,
antidiarreicos e medicamentos para controle das
comorbidades apresentadas. A cada internação os
pacientes recebem informações sobre sua doença e
a importância da adesão ao tratamento. http://www.revistas.usp.br/rmrp / http://revista.fmrp.usp.br Intencional Fonte: Adaptado de Sewitch et al. (2003)19 Discussão No presente estudo, a não-adesão ao trata-
mento medicamentoso por via oral durante o perío-
do em que permaneciam em seus domicílios ocor-
reu em 60% dos pacientes com SIC, sendo que este
comportamento foi intencional na maioria dos ca-
sos. Ainda que alguns autores estabeleçam limites
mais elevados de adesão,20 o cumprimento de 80%
das recomendações classifica o paciente como ten-
do uma elevada adesão ao tratamento.21 Considera-
se que taxas de adesão ao tratamento medicamen-
toso em torno de 50 a 60% não garantem o controle
clínico de variáveis específicas.15,16 Neste contexto,
a baixa adesão aos medicamentos nos pacientes do Fármacos e suplementos vitamínicos e
minerais de uso contínuo não adesão “intencional”, dois pacientes desconti-
nuam o uso da medicação quando se sentiam bem,
e três descontinuam a terapia quando se sentiam
mal após o uso do medicamento. Dentre os medicamentos de uso contínuo uti-
lizados pelos pacientes, os mais comuns foram os
suplementos vitamínicos e minerais, os inibidores
de bomba de prótons (omeprazol), antidiarréicos
(loperamida), fármacos para desordens gastroin-
testinais funcionais (dimeticona), sedativos
(clonazepam), várias classes de antihipertensivos,
além dos agentes antitrombóticos (varfarina sódica). Tratamento estatístico Classificação da natureza dos comportamentos em relação à não adesão ao tratamento medica-
mentoso
Natureza do comportamento
Comportamentos
Não intencional
Esquecer de tomar o medicamento
Ser descuidado quanto ao horário de tomar o medicamento
Intencional
Deixar de tomar o medicamento quando se sente bem
Deixar de tomar o medicamento quando se sente mal
Fonte: Adaptado de Sewitch et al. (2003)19 Quadro 1. Classificação da natureza dos comportamentos em relação à não adesão ao tratamento medica-
mentoso
d
C http://www.revistas.usp.br/rmrp / http://revista.fmrp.usp.br Tratamento estatístico Os dados foram tabulados em planilha Excel®
e a análise estatística descritiva foi feita com o au-
xílio do software Statistica (versão 8.0; StatSoft Inc,
Tulsa, OK, USA). Resultados Tabela 1: Dados clínicos e demográficos dos pacientes com Síndrome do Intestino Curto dependentes de
nutrição parenteral. Tempo de
Extensão
Causa da
ressecção
do intestino
Outros
Idade
IMC
ressecção
intestinal
remanescente
diagnósticos
P
(anos)
Gênero
(kg/m2)
intestinal
(meses)
(cm)
clínicos
1
63
M
16,3
Isquemia mesentérica
95
10
HAS, IRC, litiase renal
2
44
M
13,6
Trauma abdominal
36
100
HAS, insuficiência pan-
creática, litíase renal
3
69
M
23,4
Isquemia mesentérica
18
25
Trombofilia a esclarecer
4
36
F
16,1
Isquemia mesentérica
25
15
Trombofilia a esclarecer
5
50
F
18,4
Isquemia mesentérica
101
20
Trombofilia a esclarecer
6
63
F
18,8
Isquemia mesentérica
14
180
HAS, colecistite crônica,
aneurisma de aorta
7
47
M
15,7
Complicação de cirurgia
abdominal
6
60
Doença de Crohn
8
48
F
14,4
Isquemia mesentérica
12
25
Trombofilia a esclarecer
9
71
M
17,6
Isquemia mesentérica
4
30
Arritmia cardíaca,
sequela de AVE, IRC
10
71
M
20,3
Isquemia mesentérica
140
30
IRC
HAS, Hipertensão arterial sistêmica; IRC, insuficiência renal crônica; AVE, acidente vascular encefálico Tabela 1: Dados clínicos e demográficos dos pacientes com Síndrome do Intestino Curt
nutrição parenteral. nicos e demográficos dos pacientes com Síndrome do Intestino Curto dependentes de : Dados clínicos e demográficos dos pacientes com Síndrome do Intestino Curto depen
parenteral 431 Ferreira IML, Braga CBM, Dewulf NLS, Marchini JS, Cunha SFC. Baixa adesão à medicamentos na Síndrome do Intestino Curto. Medicina (Ribeirão Preto. Online) 2016;49(5):429-34 Ferreira IML, Braga CBM, Dewulf NLS, Marchini JS, Cunha SFC. Baixa adesão à medicamentos na Síndrome do Intestino Curto. Quadro 1. Classificação da natureza dos comportamentos em relação à não adesão ao tratamento medica-
mentoso
Natureza do comportamento
Comportamentos
Não intencional
Esquecer de tomar o medicamento
Ser descuidado quanto ao horário de tomar o medicamento
Intencional
Deixar de tomar o medicamento quando se sente bem
Deixar de tomar o medicamento quando se sente mal
Fonte: Adaptado de Sewitch et al. (2003)19 Quadro 1. Adesão aos medicamentos O Teste de Morisky revelou que apenas 40%
dos pacientes foram classificados como “mais ade-
rentes” (Tabela 2). A baixa condição econômica foi
relatada como um dos fatores limitantes principais
para aquisição da medicação, especialmente em
relação à aquisição do suplemento polivitamínico e
polimineral. Entre os pacientes que apresentavam Tabela 2: Adesão ao tratamento medicamentoso e natureza do comportamento relativo a não-adesão de
pacientes com Síndrome do Intestino Curto dependentes de nutrição parenteral. Paciente
Classificação da adesão
Natureza do comportamento
1
Menos aderente
Intencional
2
Menos aderente
Não intencional + intencional
3
Mais aderente
-
4
Mais aderente
-
5
Menos aderente
Não intencional + intencional
6
Menos aderente
Não intencional + intencional
7
Mais aderente
-
8
Menos aderente
Não intencional
9
Mais aderente
-
10
Menos aderente
Intencional Tabela 2: Adesão ao tratamento medicamentoso e natureza do comportamento relativo a não-adesão de
pacientes com Síndrome do Intestino Curto dependentes de nutrição parenteral. Não intencional Intencional 432 Medicina (Ribeirão Preto. Online) 2016;49(5):429-34 Ferreira IML, Braga CBM, Dewulf NLS, Marchini JS, Cunha SFC. Baixa adesão à medicamentos na Síndrome do Intestino Curto. presente estudo possivelmente influenciou no esta-
do nutricional e no controle da diarréia durante a
permanência do paciente em seu domicílio. cionais (2%) e ambos (25%).26 Os dados do pre-
sente estudo estão de acordo com estudos condu-
zidos na realidade brasileira, apesar da não-adesão
intencional ser mais prevalente em nosso estudo
que o comportamento não intencional. É possível
que o conhecimento do paciente sobre o compro-
metimento da capacidade absortiva induza ao ceti-
cismo quanto à eficácia da terapêutica oral, razão
pela qual a não-adesão dos pacientes com SIC te-
nha sido intencional. No início de um programa de educação ali-
mentar, os pacientes são resistentes às mudanças
propostas, principalmente em relação ao fraciona-
mento da dieta.22 Entretanto, ao se conscientiza-
rem da importância e benefícios alcançados com a
dieta, a adesão torna-se mais elevada.22 A baixa
adesão ao planejamento dietético foi ilustrada em
relato de caso de um paciente com SIC que desen-
volvia quadros recorrentes de acidose metabólica.23 Thyssen e colaboradores (2014)3 sugerem que
o ambiente de convívio do paciente é essencial para
garantir sua participação nos seus cuidados e tra-
tamentos. O perfil socioeconômico da nossa casu-
ística pode ter contribuído para a baixa adesão ao
tratamento medicamentoso. Adesão aos medicamentos A Organização Mundi-
al da Saúde documentou que em países desenvol-
vidos a não-adesão ao tratamento é em torno de
50% para condições crônicas e que esse cenário
seria ainda maior em países em desenvolvimento.17
Em geral, são necessárias múltiplas intervenções
para que mudanças comportamentais modestas
sejam alcançadas.14,15,16 A observação em nosso
estudo de que a não-adesão ‘intencional’ foi mais
prevalente nos paciente com SIC ressalta a neces-
sidade de intervenções educativas que mostrem a
importância da adesão ao tratamento medicamen-
toso.27 As estratégias visando melhoria da adesão
ao tratamento devem incluir a educação básica em
saúde, estabelecimento de metas factíveis e a cria-
ção de grupos de apoio social ou profissional.14 O
acompanhamento telefônico pode contribuir para a
elevação da adesão ao tratamento medicamentoso
experimental em pacientes com SIC,9,23 no trata-
mento da asma e do tabagismo.17 Apesar da suplementação oral no âmbito do-
miciliar, os pacientes com SIC reduziram os níveis
séricos de tocoferol no período entre os ciclos de
nutrição parenteral5. Embora a hipótese mais pro-
vável seja que a baixa capacidade absortiva, a não
adesão à suplementação oral de vitaminas pode ter
contribuído com os resultados obtidos. Dois estu-
dos dinamarqueses foram conduzidos para investi-
gação da capacidade absortiva de pacientes com
SIC após o uso parenteral com peptídeo semelhan-
te ao glucagon.9,24 Paralelamente, os autores avali-
aram a adesão ao tratamento por métodos indire-
tos que incluíram a contagem das ampolas vazias,
o registro diário dos efeitos colaterais e os relatos
do não cumprimento da prescrição. Nos 35 dias ini-
ciais do estudo, oito entre os 11 indivíduos com SIC
apresentaram adesão ao tratamento superior a
94%.24 Quando o tempo da terapia se prolongou
por dois anos, cerca de 30% dos voluntários não
mantiveram elevado nível de adesão ao tratamen-
to.9 A avaliação da adesão ao tratamento medica-
mentoso pode variar em função do método empre-
gado no estudo, do nível socioeconômico e cultural
da população, além da doença de base. No presen-
te estudo, o método de avaliação da adesão ao tra-
tamento foi o mesmo empregado em pesquisas bra-
sileiras conduzidas em pacientes com doenças in-
flamatórias intestinais.25,26 Em nosso meio, 50% dos
pacientes com doença de Crohn apresentaram bai-
xa adesão ao medicamento, sendo que a não-ade-
são não-intencional ocorreu em 42% dos casos. http://www.revistas.usp.br/rmrp / http://revista.fmrp.usp.br Ferreira IML, Braga CBM, Dewulf NLS, Marchini JS, Cunha SFC.
Baixa adesão à medicamentos na Síndrome do Intestino Curto. Referências 19. Sewitch MJ, Abrahamowicz M, Barkun A, Bitton A, Wild GE,
Cohen A. Patient Nonadherence to Medication in Inflam-
matory Bowel Disease. Am J Gastroenterol. 2003; 98:1535-
44. 1. O’Keefe SJ, Buchman AL, Fishbein TM, Jeejeebhoy KN,
Jeppesen PB, Shaffer J. Short bowel syndrome and intes-
tinal failure: consensus definitions and overview. Clin
Gastroenterol Hepatol. 2006; 4: 6-10. 20. Pullar T, Kumar S, Tindall H, Feely M. Time to stop counting
the tablets? Clin Pharmacol Ther. 1989; 46: 163-8. 2. Tappenden KA. Pathophysiology of short bowel syndrome:
considerations of resected and residual anatomy. JPEN J
Parenter Enteral Nutr. 2014; 38(1 Suppl):14S-22S. 21. Rudd P, Byyny RL, Zachary V, Loverde ME, Titus C, Mitchell
WD. The natural history of medication compliance in a drug
trial: limitations of pill counts. Clin Pharmacol Ther. 1989;
46: 169-76. 3. Thyssen GD, Beck A. How patients experience the sur-
roundings in relations to patient participation: a qualita-
tive study of inpatients with intestinal failure. Patient Prefer
Adherence. 2014; 8: 585-92.101014 22. Nonino CB, Borges RM, Pasquali LS, Marchini JS. Terapia
nutricional oral em pacientes com síndrome do intestino
curto. Rev Nutr. 2001; 14: 201-5. 23. Gilchrist PN, Phillips PJ, Heddle R, Harley H. Dietary com-
pliance in the short bowel syndrome. JPEN J Parenter En-
teral Nutr. 1984; 8: 315-6. 4. Braga CBM, Vannucchi H, Freire CM, Marchini JS, Jordão
AA Jr, da Cunha SF C. Serum vitamins in adult patients
with short bowel syndrome receiving intermittent
parenteral nutrition. JPEN J Parenter Enteral Nutr. 2011;
35: 493-8. 24. Jeppensen PB, Hartmann B, Thulesen J, Graff J, Lohmann
J, Hansen BS, et al. Glucagon-like peptide 2 improves nu-
trient absorption and nutritional status in short-bowel pa-
tients with no colon. Gastroenterology. 2001; 120: 806-
15. 5. Ferreira IM, Braga CBM, Dewulf NL, Marchini JS, da Cunha
SFC. Vitamin serum level variations between cycles of in-
termittent parenteral nutrition in adult patients with short
bowel syndrome. JPEN J Parenter Enteral Nutr. 2013; 37:
75-80. 25. Dewulf NLS, Monteiro RA, Passos ADC, Vieira EM, Troncon
LEA. Adesão ao tratamento medicamentoso de pacientes
com doenças inflamatórias intestinais acompanhados no
ambulatório de um hospital universitário. Arq Gastroenterol. 2007; 44: 289-96. 6. Braga CBM, Ferreira IM, Marchini JS, da Cunha SFC. Cop-
per and magnesium deficiencies in patients with short
bowel syndrome receiving arenteral nutrition or oral feed-
ing. Arq Gastroenterol. 2015; 52:94-9. 26. Adesão aos medicamentos Entre os pacientes com retocolite ulcerativa ou colite
indeterminada, 63% apresentavam baixa adesão ao
tratamento medicamentoso e a não-adesão foi clas-
sificada em não-intencional em 60% dos pacien-
tes.25 Entre 100 pacientes com doença de Crohn,
64% apresentaram escore compatível com a não-
adesão por motivos não-intencionais (37%), inten- Dentre as limitações do estudo, pode-se in-
cluir o pequeno tamanho amostral. No entanto, com-
parando-se o número de casos estudados em ou-
tras pesquisas envolvendo pacientes com SIC3,28,29
e sua baixa prevalência mundial, pode-se afirmar
que o tamanho da amostra foi relativamente ex-
pressivo. O questionário usado neste estudo não é
específico para pacientes com SIC e por isso não
inclui questões direcionadas às queixas comumen-
te observadas na síndrome. São necessários estu-
dos que investiguem a adesão medicamentosa por
outros métodos de avaliação e as principais causas
de não-adesão em pacientes com SIC. A adesão ao
tratamento deve ser encorajada persistentemente
nos pacientes com SIC, considerando a alta preva- 433 Medicina (Ribeirão Preto. Online) 2016;49(5):429-34 Ferreira IML, Braga CBM, Dewulf NLS, Marchini JS, Cunha SFC. Baixa adesão à medicamentos na Síndrome do Intestino Curto. Ferreira IML, Braga CBM, Dewulf NLS, Marchini JS, Cunha SFC. Baixa adesão à medicamentos na Síndrome do Intestino Curto. ment: adherence, adaptation, and practical recommenda-
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ating successful self-care plans. Am J Nurs. 2007; 107: 20-
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documentos que a maioria dos pacientes tem baixa
adesão ao tratamento medicamentos por via oral
durante o período em que eles permanecem em seus
domicílios. Nossos resultados sugerem o desenvol-
vimento e a implementação de estratégias que vi-
sem à percepção da relevância do uso correto dos
medicamentos e a promoção da adesão ao trata-
mento medicamentoso. 15. Moriksy DE, Green LW, Orisky DE, Green LW, Levine DM. Concurrent and predictive validity of a self-reported meas-
ure of medication adherence. Med Care. 1986; 24: 67-74. 16. Morisky DE, Ang A, Krousel-Wood M, Ward HJ. Predictive
validity of a medication adherence measure in an outpa-
tient setting. J Clin Hypertens. (Greenwich). 2008; 10: 348-
54. 17. WHO. World Health Organization. Adherence to long-term
therapies: evidence for action. 2003, Geneva: World Health
Organization. 18. Adesão aos medicamentos Garber MC, Nau DP, Erickson SR, Aikens JE, Lawrence JB. The concordance of self-report with other measures of
medication adherence. Med Care. 2004; 42: 649-52. Referências Cornélio RCAC, Pinto ALT, Pace FHL, Moraes JP, Chebli JMF. Não-adesão ao tratamento em pacientes com doença de
Crohn: prevalência e fatores de risco. Arq Gastroenterol. 2009, 46: 183-9. 7. Buchman AL, Scolapio J, Fryer J. AGA technical review on
short bowel syndrome and intestinal transplantation. Gas-
troenterology. 2003; 124: 1111-34. 27. Wang MY, Wu MH, Hsieh DY, Lin LJ, Lee PH, Chen WJ, et al. Home parenteral nutrition support in adults: experience of
a Medical Center in Asia. JPEN J Parenter Enteral Nutr. 2007;
31: 306-10. 8. Storch KJ. Overview of Short Bowel Syndrome: Clinical
Features, Pathophysiology, Impact, and Management. JPEN J Parenter Enteral Nutr. 2014; 38 (1 Suppl): 5S-7S. 9. Jeppensen PB, Lund P, Gottschalck IB, Nielsen HB, Holst
JJ, Mortensen J, et al. Short bowel patients treated for
two years with glucagon-like peptide 2 (GLP-2): compli-
ance, safety, and effects on quality of life. Gastroenterol
Res Pract. 2009; 2009:425759: 1-9. 28. Atia A, Girard-Pipau F, Hébuterne X, Spies WG, Guardiola
A, Ahn CW et al Macronutrient absorption characteristics
in humans with short bowel syndrome and jejunocolonic
anastomosis: starch is the most important carbohydrate
substrate, although pectin supplementation may modestly
enhance short chain fatty acid production and fluid ab-
sorption. JPEN J Parenter Enteral Nutr. 2011; 35: 229-40. 10. Matarese LE, Steiger E. Dietary and medical management
of short bowel syndrome in adult patients. J Clin
Gastroenterol. 2006; 40 (Suppl 2): S85-S93. 29. Seguy D, Darmaun D, Duhamel A, Thuillier F, Cynober L,
Cortot A, et al. Growth hormone enhances fat-free mass
and glutamine availability in patients with short-bowel
syndrome: an ancillary double-blind, randomized crosso-
ver study. Am J Clin Nutr. 2014; 100: 850-8. 11. Kumpf VJ. Pharmacologic Management of Diarrhea in Pa-
tients With Short Bowel Syndrome. JPEN J Parenter Enteral
Nutr. 2014; 38 (1 Suppl): 38S-44S. 12. Vantini I, Benini L, Bonfante F, Talamini G, Sembenini G,
Chiarioni G, et al. Survival rate and prognostic factors in
patients with intestinal failure. Dig Liver Dis. 2004; 36:
46-55. 13. Wall EA. An overview of short bowel syndrome manage- http://www.revistas.usp.br/rmrp / http://revista.fmrp.usp.br 434
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Isabel Lincoln Strange Reséndiz
La irrupción de lo audiovisual en los procesos
sociales: una reflexión a partir de la teoría
Revista Xihmai XV (29), 83-104, enero-junio 2020
Universidad La Salle Pachuca
xihmai@lasallep.edu.mx
Teléfono: 01(771) 717 02 13 ext. 1406 Fax:
01(771) 717 03 09
ISSN (versión impresa):1870-6703 México.
https://doi.org/10.37646/xihmai.v15i29.336
2020
Isabel Lincoln Strange Reséndiz
LA IRRUPCIÓN DE LO AUDIOVISUAL EN LOS PROCESOS
SOCIALES: UNA REFLEXIÓN A PARTIR DE LA TEORÍA
THE IRRUPTION OF THE AUDIOVISUAL IN SOCIAL
PROCESSES: A REFLECTION BASED ON THEORY
Xihmai, año 2020/vol. XV, número 29
Universidad La Salle Pachuca
pp. 83-104
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Revista Xihmai XV (29), 83-104, enero-junio 2020
LA IRRUPCIÓN DE LO AUDIOVISUAL EN LOS PROCESOS
SOCIALES: UNA REFLEXIÓN A PARTIR DE LA TEORÍA
THE IRRUPTION OF THE AUDIOVISUAL IN SOCIAL
PROCESSES: A REFLECTION BASED ON THEORY
Isabel Lincoln Strange Reséndiz
Doctora en Ciencias Políticas y Sociales,
línea Comunicación y Cultura,
por la Facultad de Ciencias Políticas y Sociales
de la UNAM. Profesora Investigadora de la Licenciatura
en Comunicación y el Centro de Investigación
en Comunicación Aplicada (CICA)
de la Universidad Anáhuac Norte.
isabelincoln@hotmail.com
Ante el fracaso de la ideología racionalista del progreso lineal
y continuo, la comunicación ha tomado el relevo y se presenta
como parámetro por excelencia de la evolución de la humanidad,
en un momento histórico en el que ésta busca desesperadamente,
un sentido a su futuro
(Mattelart y Mattelart, 2003, p. 125).
La televisión puede así convertirse en instrumento eficaz
para una acción de pacificación y de control, en garantía
de conservación del orden establecido
a través de la repetición de aquellas opiniones
y de aquellos medios que la clase dominante juzga
más aptos para mantener el status quo
(Eco, 2006, p. 354).
Resumen
El presente artículo analiza la irrupción de lo audiovisual en el estudio de la
comunicación a partir del surgimiento de la televisión y hasta su condición
actual. El objetivo de este texto consiste en observar el impacto de los medios
audiovisuales en los procesos sociales. Para ello, se realizó una revisión de las
propuestas teóricas que expusieran los procesos históricos y la repercusión que
la comunicación tuvo en los mismos. Al final de este documento, se reflexiona
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Revista Xihmai XV (29), 83-104, enero-junio 2020
en el transmedia, la condición actual del receptor y su relación con algunos
productos, tomando como ejemplo para ello, la película Black Mirror:
Bandersnatch (David Slade, 2018).
Palabras clave: comunicación, televisión, audiovisual, transmedia, cultura.
Abstract
This article analyzes the emergence of the audiovisual in the study of
communication, from the emergence of television to its current condition. The
objective of this text is to observe the impact of audiovisual media on social
processes. For this, a review of the theoretical proposals that exposed the
historical processes and the impact that communication had on them was
carried out. At the end of this document, the current condition of the
transmedia, the receptor and its relationship with some products are reflected,
taking as an example, the film Black Mirror: Bandersnatch (David Slade,
2018).
Keywords: Communication, television, audiovisual, transmedia, culture.
Una perspectiva histórica de la comunicación a partir de la teoría
La evolución de los productos audiovisuales está relacionada con los procesos
históricos; esta es una cuestión que han observado diversos pensadores a lo
largo de los años. Uno de ellos es el autor belga Armand Mattelart, uno de los
principales representantes de la corriente teórica denominada Mass Media
Comunication Reasearch, que destaca la importancia que tienen los medios de
comunicación en la sociedad, las políticas de Estado y, en consecuencia, los
procesos históricos.
El enfoque de Armand Mattelart sobre la internacionalización de la
comunicación nos permite comprender el hecho de que los procesos de
comunicación y los procesos históricos están entrelazados: unos dependen
directamente de otros. Las propuestas del teórico refuerzan los argumentos
expuestos por las diversas escuelas de los estudios de la comunicación en el
sentido en que productos culturales y audiovisuales tienen un fuerte impacto
en nuestro aprendizaje.
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La mundialización en la comunicación (1996) sigue línea de estudio histórica,
política y cultural. Para Mattelart, la internacionalización de la comunicación
sucede a partir de dos universalismos, la Ilustración y el liberalismo,
específicamente. En el primero, el surgimiento de la enciclopedia y los autores
del contexto, como Diderot, Voltaire, Montesquieu, entre otros, pusieron en
circulación la pertinencia de la comunicación como un ideal de la modernidad,
así como la premisa del libre tránsito de las ideas y de las opiniones. Desde el
punto de vista de Mattelart, el triunfo de la Revolución Francesa representó la
universalización y la liberalización de los intercambios económicos y políticos;
esta idea que se hizo popular en el mundo entero y sirvió de apoyo para otro
tipo de movimientos internacionales que buscaron la libertad, no solo de sus
gobiernos, sino de sus formas de vida, sus ideas y sus expresiones (1996, pp.
11-14).
La aparición de objetos que agilizaron las comunicaciones, como el telégrafo
óptico, permitieron que “la técnica de comunicación a larga distancia fuera
considerada garante de una democracia renovada” (Mattelart, 1996, p. 12). Los
avances subsecuentes en la sociedad francesa, como la división del trabajo en
el ámbito internacional, establecieron nuevas formas de comunicación; la
expansión del ferrocarril durante el siglo XIX y el telégrafo transatlántico
impulsaron la comunicación en una escala casi global.
El descubrimiento de nuevos medios (formas) de comunicación ha sido una
constante desde la Revolución Industrial. El impulso de las
radiocomunicaciones trasatlánticas y la regulación de las mismas, por ejemplo,
fue esencial: “ha existido una estrecha relación entre el desarrollo de las
tecnologías de la comunicación y los conflictos que se desarrollaron en la
segunda mitad del siglo XIX” (Mattelart, 1996, p. 21). El expansionismo de
los medios de comunicación en el mundo entero fue impulsado por las grandes
potencias:
Antes del estallido de la Primera Guerra Mundial se producen dos guerras: la guerra
de los Boers (1899-1902) y la guerra ruso-japonesa (1904-1905); donde se
confirma, en el caso de la primera, el peso decisivo del tren y del telégrafo y, en la
segunda, el papel que desempeñan las radiocomunicaciones. No tardará Inglaterra
en extraer las enseñanzas de este conflicto, declarando la radiotelegrafía monopolio
estatal […] (Mattelart, 1996, p. 22).
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La comunicación, o más precisamente las redes de comunicación, fue una
justificación importante para la colonización debido a que los medios dictaron
la necesidad de tender redes de cableado, antenas y otro tipo de infraestructura.
Esto es ejemplo de lo que Mattelart denomina las “utopías de la comunicación
universal”; si bien se puede considerar que un planeta comunicado representa
una utopía, nueva organización universal con menos conflictos, en realidad
existen intereses relacionados con la administración de la comunicación que,
ciertamente, significan nuevos retos para los problemas de comunicación, en
la medida en que existen intereses (económicos y políticos) relacionados con
los mismos y que son controlados por los grupos que están en el poder:
En 1881, se celebró en París la primera exposición internacional de la electricidad.
Los delegados de las potencias poseedoras de las patentes ligadas a esta invención
se reunieron en el marco de este acontecimiento […] ningún Estado soberano de la
periferia sería invitado a esta reunión en la cumbre (Mattelart, 1996, p. 28).
En el siglo XX, los medios de comunicación adquirieron nuevas dinámicas de
funcionamiento debido a que la industrialización alcanzó todos los aspectos de
la misma. La prensa, por ejemplo, se transformó en un medio industrial.
Mattelart señala que las primeras plataformas diseñadas para analizar la
economía internacional compartían su información, de manera paralela, con la
prensa: “la empresa J. Walter Thompson, por ejemplo, trabajaba en Estados
Unidos, Francia y Londres” (Mattelart, 1996, p. 32).
Imagen 1. Publicidad de la empresa J. Walter Thopson
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La prensa industrializada es un fenómeno que se observará en varios países,
no solo como resultado de la presencia de las agencias sino de un cambio en la
estructura del medio. Durante casi todo el siglo XX, la prensa fue empresarial
e industrial; los diarios y las revistas contaban con varias secciones, cada vez
más definidas según los intereses de los lectores y dedicadas a ampliar las
ventas de las compañías editoras. Desde el punto de vista de Mattelart, incluso
la literatura por entregas se convirtió una “literatura industrial” (Mattelart,
1996, p. 33); lo mismo sucedió con las secciones de historietas, publicidad o
moda; cada una de estas iba enfocada a un tipo de público. La prensa estuvo
dirigida a diversos sectores: “En vísperas de la Primera Guerra Mundial, el
debate sobre la naturaleza del público, así como lo que constituye su corolario,
esto es, el poder de persuasión de la prensa sobre los lectores, está dominado
por una tendencia llamada fusionista; según esta, la proyección parte siempre
de un centro tutelar que impone su visión del mundo a las distintas periferias”
(Mattelart, 1996, p. 37).
En este sentido, debemos reflexionar en el hecho de que la Primera Guerra
provocó el surgimiento de un nuevo tipo de propaganda. Por ejemplo, en el
caso de Alemania, en 1917 se creó la Universum Film AG, una compañía
productora de películas en la que participaron todos los sectores de la industria
del país y que, además, fue apoyada por empresarios alemanes. Universum
Film se creó con el propósito de controlar el país, a través de una industria
cinematográfica que mantuviera el mercado interno, aunado a la política de
prohibición de importación de filmes.
La crisis económica que la Primera Guerra Mundial ocasionó en Europa fue
aprovechada por los Estados Unidos. En el caso del cine, por ejemplo, si bien
la compañía Pathé siguió funcionando, la mayor parte de los filmes que se
veían en Europa provenían de los Estados Unidos, por lo que, a finales de la
década de los veinte, algunos países como Francia obligaron a los exhibidores
a proyectar de manera obligatoria entre un treinta y un cincuenta por ciento de
filmes nacionales. La industria cinematográfica, como es bien sabido, despegó
en el país del norte durante los años inmediatos posteriores a la Gran Guerra y
su éxito se mantiene hasta nuestros días.
En este sentido, es necesario mencionar que la publicidad ha sido fundamental.
Mattelart menciona que la publicación de los libros Public Opinion de Walter
Lippmann y Propaganda Techniques in the War World de Harold Laswell
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fueron obras fundamentales para comprender el impacto que esta tuvo en las
relaciones comerciales, políticas y sociales. Los medios de comunicación se
promovían mutuamente y las grandes industrias hacían uso de estos medios:
En 1924, se produjo un acercamiento entre la asociación de agencias de publicidad
británicas y su homóloga en los Estados Unidos. En 1938, se creó en Nueva York
la International Advertising Association […] Con estas ideas llega también otra idea
que establece un lazo entre la democracia y la Democratic Market Place: la libertad
de expresión de los ciudadanos y la libertad de expresión comercial (Mattelart,
1996, p. 50).
La publicidad y la propaganda permitieron –y han permitido– manejar la
ideología de las naciones. No olvidemos que la radio también se convirtió en
un medio fundamental después de la Primera Guerra Mundial. El invento, que
suele concederse a Tesla, Marconni o Hertz, fue un medio que permitió la
comunicación instantánea desde las primeras décadas del siglo XX.
Recordemos que obtuvo reconocimiento internacional después del
hundimiento del Titanic debido a que fue mencionado, de manera constante,
durante las investigaciones. Durante la Gran Guerra, la radio permitió la
comunicación entre los ejércitos europeos e, incluso, con los Estados Unidos.
Según apunta Raymond Williams, en sus inicios, la radio se presentó como un
medio accesible y de fácil distribución, además de presentar un contenido
variado de noticias, opinión y teatro. La tecnología ocasionó un cambio radical
y una evolución acelerada de los medios de comunicación: “Las habilidades
sociales más básicas, de un tipo adquirido en un desarrollo y una relación más
bien primarios, dieron acceso al cine, a la radiodifisión, a los programas de
televisión” (1992, pp. 205-206).
La radio propició el avance hacia la internacionalización de la comunicación.
En 1923, durante el gobierno de Lenin, en la Unión Soviética se creó la
Agencia de Prensa Oficial, que mantenía el control de la prensa, el cine y la
radio. Asimismo, en Alemania, una vez que el Partido Nacional Socialista
Obrero Alemán llegó al poder, también se creó una estrategia de control
ideológico que operaba a través de estos medios. La historia se repite en otras
naciones. En 1925, en Estados Unidos se creó la Unión Internacional de
Radiodifusión; en 1931, Radio Vaticano inició sus transmisiones. También
Gran Bretaña, a través de la BBC, en ese entonces Britsh Broadcasting
Company Ltd, inició sus transmisiones e incluso, durante los años treinta y
frente a la amenaza nazi-fascista en Europa, “creó un servicio en lengua
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alemana, difundido en español y portugués en América Latina” (Mattelart,
1996, p. 53). La radio, junto con la prensa y el cine, serían los trasmisores de
todo tipo de ideologías durante el periodo de entre guerras y, sin duda, serían
fundamentales durante la Segunda Guerra Mundial.
Raymond Williams apunta que los usos principales de las nuevas tecnologías
dependieron de grandes capitales de comunicación y establecieron relaciones
complejas con las instituciones más comunes, como el Estado, la iglesia, la
escuela y la familia, y contribuyeron en la preparación social y cultural (1992,
p. 207).
En Alemania, el tercer Reich mantuvo el control de casi todo el proceso de
comunicación, independientemente del medio que se tratase. Los mensajes,
obviamente, estaban destinados a respaldar la ideología del Estado; con la
distribución de los radios Volksempfänger, “Voz del pueblo”, y las
transmisiones de la Reichs-Rundfunk-Gesellschaft, las radiodifusoras locales,
Goebbels pudo poner en marcha su Ministerio para la ilustración pública y
propaganda. No obstante, el mensaje de la propaganda era variado ya que la
programación contenía tanto noticias oficiales de lo que sucedía en Alemania
como conciertos, programas culturales y de entretenimiento.
No obstante, el medio de comunicación que mantuvo su supremacía sobre los
demás fue la radio, aunque los tres compartían el éxito. Después de la Segunda
Guerra, varios países de Europa pusieron en marcha radiodifusoras estatales y,
al mismo tiempo, surgieron las llamadas “clandestinas”. El fenómeno de las
radios no autorizadas tuvo un gran impacto en Europa; surgieron este tipo de
emisoras a partir de los años cincuenta como Radio Free Europe, Radio
Liberty, Radio Caroline; esta última transmitía desde una embarcación en
altamar durante la década de los sesenta.
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Imagen 2.- Roosevelt en la Segunda Guerra Mundial.
En este triángulo en el que la radio, la prensa y el cine compartían el éxito, la
televisión comercial entró en escena. Si bien en los inicios de este medio las
dos principales empresas televisivas, la BBC (Gran Bretaña) y la CBS (Estados
Unidos), tenían sistemas distintos (estatal y comercial), ambas estaban
condicionadas por intereses económicos, comerciales e ideológicos.
Por otro lado, durante las primeras décadas del siglo XX, el cine se consolidó
como uno de los principales medios masivos de comunicación. Desde su
surgimiento en 1895, avanzó a pasos agigantados, evolucionando sus formas
narrativas y aprovechando las innovaciones tecnológicas. Ambos aspectos, el
desarrollo tecnológico y la consolidación de su lenguaje, tuvieron un
importante impacto social. El acorazado Potemkin (Eisenstein, 1925) fue una
de las cintas más representativas en los primeros años del Estado Soviético.
Según apunta F. Albèra (1998), “Las directrices pedagógicas dictadas por
Lenin en el momento de la nacionalización de la industria cinematográfica son
superadas en la cinta” (p. 7). De esta manera, el cine se convirtió en un medio
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de control ideológico y, a la vez, en una de las principales formas de arte y
entretenimiento. Sus avances a lo largo del siglo XX, como sonido, color y
efectos especiales, así como su distribución y presentación, han determinado
su relación con el espectador, incluso en nuestro días, como se verá más
adelante.
Imagen 3. Cartel de El acorazado Potemkin
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Raymond Williams (1992) señala que hacia la década de los cincuenta sucede
un cambio cualitativo en la distribución de los procesos de comunicación: “el
suministro equitativo a los medios de comunicación directamente determinada
que sirviera a necesidades inmediatas individuales y sociales” (p. 209). Se trata
de aspectos que están relacionados con políticas de distribución.
Hacia la década de los cincuenta, la televisión modificó la forma de ver el
mundo, casi siempre de manera condicionada por una perspectiva
mercantilista. Desde la incursión de este medio en la vida cotidiana, la
televisión condicionó la vida de las relaciones sociales. Si bien la
conformación de los primeros programas de televisión guardaba una estrecha
relación con otros medios de comunicación, como el cine y la radio, esta
estableció sus propias dinámicas de funcionamiento y su propio lenguaje.
La televisión se mantuvo como uno de los medios más importantes en el
mundo gracias, en gran medida, a los avances tecnológicos subsecuentes, como
la comunicación satelital, que permitió el crecimiento económico de grandes
compañías: “La palabra transnacional implica la existencia de un movimiento
de conjunto hacia la integración a nivel mundial, y viene a significar que existe
una fuente de conflictos entre los intereses de las macro empresas y los
territorios en las que éstas se asientan” (Mattelart, 1996, p. 67). Empero, estos
mismos países de primer mundo, que anteriormente habían resguardado sus
intereses particulares, fueron los que denunciaron que las comunicaciones no
se daban de la misma manera para todos los seres humanos; la integración de
los países de la “periferia” al libre y fácil intercambio de comunicación de
calidad (educación, información, cultura), según apunta el autor, permitiría
mejores condiciones de vida para la humanidad en general.
También a principios de la década de los cincuenta los estudios de la Escuela
de Frankfurt y sus representantes, Max Horkheimer (La personalidad
autoritaria), Theodor W. Adorno (Dialéctica de la Ilustración), H. Marcusse
(El hombre unidimensional), pusieron de manifiesto que el arte y la cultura se
legitiman a partir de nuevas propuestas ideológicas. Sus análisis en torno a la
industria cultural subrayan la importancia que tiene el contexto histórico y su
ideología en los productos audiovisuales. Una década después, en 1964, el
surgimiento de la Escuela de Birmingham, con autores como Richard Hoggart
(The Uses of Literacy) y Stuart Hall (La cultura y el poder) pusieron en
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perspectiva la relación entre la condición de clase, el grupo social, el contexto
histórico, cultura y medios.
En una conferencia inaugural, en 1964, Hoggart planteó la óptica inicial del centro.
Se trataba fundamentalmente de movilizar las herramientas y técnicas de la crítica
literaria para desplazarlos hacia temas que, hasta entonces, eran considerados
ilegítimos por la comunidad universitaria: el universo de las culturas y las prácticas
populares en oposición a las culturas letradas, la toma en cuenta de la diversidad de
bienes culturales que abarcará los productos de la cultura de los medios de
comunicación social, después de los estilos de vida, y ya no sólo las obras literarias
(Mattelart et al., 2002, p. 34).
Por otro lado, dentro de la óptica de Apocalípticos e integrados, Umberto Eco
señala que la modificación de los instrumentos culturales en la historia es una
puesta en crisis de todo el modelo cultural que no manifiesta su alcance real
cuando quienes lo analizan no toman en cuenta los elementos que operan en
un contexto determinado, ya que la sociedad se ha transformado con el paso de
los años. Para Eco, la evolución de la comunicación implica una serie de
problemas relacionados con los aspectos histórico, antropológico y cultural
(2006, pp. 52-53).
Eco señala que los medios masivos de comunicación se presentan como la
herramienta educativa, típica de una sociedad paternalista, “tendiente a
producir modelos humanos heterodirigidos […] desarrollan la misma función
que en ciertas circunstancias históricas ejercieron las ideologías religiosas” (p.
59). El mensaje emitido de esta forma crea un velo entre lo que identificamos
como realidad y como ficción. No obstante, si bien los medios masivos
proponen información que puede volverse “formación”, esto no significa que
exista una homogenización en la recepción de los contenidos, debido a que
cada mensaje: “es constituyente de un conjunto de nuevos contenidos” (2006,
p. 65). Como señala el autor, se dan casos en que los medios están en manos
de grupos de poder político que quieren ejercer el dominio pero que siguen
actuando de manera industrial. La cultura de masas es producida por grupos
que mantienen el poder económico y producen cultura de masas con el fin de
obtener beneficios particulares; en este sentido, el mensaje se convierte en un
producto que entra en el mercado de la venta y la distribución.
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Las imágenes y la universalidad de la comunicación
Pareciera, a partir de lo mencionado, que existe una estandarización y
homogenización universal de los medios de comunicación y los productos
audiovisuales. La década de los ochenta y los cambios globales modificaron
las dinámicas de relación comercial:
La creación de un mercado único de imágenes constituye un reto en la búsqueda de
una cultura denominada global […] La tercera generación de redes publicitarias, las
redes denominadas globales, favorecidas por la integración de operaciones de
comunicación, respondiendo al movimiento de interconexión de los mercados […]
Las industrias culturales de Estados Unidos, soportes naturales de la universalidad,
aparecen siempre estableciendo excesivamente los parámetros de la globalidad
(Mattelart, 1996, p. 92).
Esta realidad quedó subrayada con el boom de internet en la década de los
noventa debido a que la sociedad se vio condicionada por la interconexión.
Hoy en día, esta es una realidad cotidiana para la gran parte de la población
internacional. Si bien algunas zonas del mundo se mantienen alejadas de
internet, este medio controla casi la totalidad de la comunicación. Aunque el
medio de comunicación no llegue a cada rincón de la tierra, existen intereses
económicos y políticos que obligarán a las grandes potencias a intervenir en
un espacio ajeno. La industrialización de los medios, finalmente, condiciona
relaciones políticas y sociales y, en consecuencia, tiene un impacto en la
historia.
Armand Mattelart, en Geopolítica de la cultura (2002), señala que hacia finales
del siglo XX surgió una controversia en torno a la creación cultural. Cada
época adapta este concepto según sus necesidades. Después del fin de la Guerra
Fría, en 1991, los ministerios de cultura en Europa empiezan a promover el
consumo de la llamada “alta cultura” y de sus productos culturales nacionales
para contrarrestar el impacto del crecimiento de la producción audiovisual a
nivel internacional.
A principios de la década de los noventa, en vistas de crear la Unión Europea
(1993), se expone que “no respetar la cultura es dejarla fuera de las
competencias […] es decir, es un no radical al triunfo del mercado sin
conciencia ni misericordia” (Jack Ralite citado en Mattelart, 2002, p. 77). El
autor subraya que “la idea de que existe una ineludible tendencia a la
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unificación cultural del mundo no es de hoy” (p. 84), por el contrario, como lo
hemos expuesto, el fenómeno que surge con la industrialización de la
comunicación y que está estrechamente relacionado con los procesos
históricos, a pesar de que se crearon agencias que buscaban revolucionar
contenidos, los países occidentales aún temían a la amenaza de que la ideología
socialista se infiltrara en su territorio.
En este sentido, la posibilidad de una “cultura global” a través de diversas
cadenas de televisión se apoya en la noción de la cultura de masas y en el
imaginario relacionado con que el hecho de que los consumidores sean
procedentes de culturas muy diversas establecerá una diferencia (Mattelart,
2002, p. 96); la cultura se convierte en transfronteriza. Es esencial señalar que,
a través de los productos culturales y de los procesos de comunicación que
existen en torno a ellos, las naciones se apropian, nuevamente, de su historia,
debido a que buscan la pureza de las identidades etnoculturales a través de
estos mismos procesos. Existe un intercambio constante entre lo local y lo
global en el que la televisión y los productos audiovisuales tendrán un peso
importante en el intercambio cultural. Incluso, “el Tercer Mundo ha pasado a
ocupar un lugar destacado en el mercado mundial de programas de televisión
[…] y de productos culturales que le ha permitido ampliar sus fronteras al
anexionarse los nuevos territorios del antiguo bloque comunista y por lograr
que la India y China” (Mattelart, 2002, p. 94).
Lo audiovisual y los procesos sociales contemporáneos
Elizabeth Evans, en su texto Transmedia television. Audiences, new media and
daily life, anota que, para el teórico de la comunicación audiovisual Noël
Carrol, la naturaleza ontológica de la televisión se encuentra en su relación con
el cine; la TV es un estado de evolución de la imagen movimiento y, por lo
tanto, sus atributos no tienen una única definición (2011, p. 11). De acuerdo
con la cita anterior, es posible argumentar que casi todo producto audiovisual
guarda un vínculo con la televisión como medio de comunicación.
La definición de televisión como medio de comunicación se ha ido
modificando con el paso de los años, no solamente desde el punto de vista
teórico, sino también desde el punto de vista práctico, técnico y comercial. Las
investigaciones en torno a los nuevos medios tienen sus orígenes en la
televisión. De hecho, el término televidente o receptor, generalmente utilizado
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por los teóricos, ha sido reemplazado por el de usuario, un sujeto que accede
al medio a través de un dispositivo que no es el objeto en sí. Si reflexionamos
en la definición de usuario de internet, se trata de un sujeto que es más activo
y participativo que el espectador televisivo; sus inquietudes en torno al medio
incluyen los modos y condiciones del internet, esto aunado a su relación
primigenia con la televisión.
El vuelco de lo digital ha generado una importante reflexión en torno a la
naturaleza del medio. Sin duda alguna, las audiencias responden a los cambios
por lo que ha pasado la televisión es fundamental. Evans anota que Noël
Carroll ofrece dos aproximaciones en torno a la definición de televisión: a) la
naturaleza ontológica de la televisión se encuentra en su relación con el cine;
b) es un estado de evolución de la imagen movimiento y, por lo tanto, sus
atributos no tienen una única definición (Evans, 2011, p. 3).
Los estudios académicos se han enfocado en dos vertientes relacionadas con la
televisión: su uso doméstico y familiar; los usos subculturales de programas
específicos de la televisión para ciertos grupos sociales; la relación de los fans
con sus series de televisión; el impacto de la tecnología en el contenido
televisivo (Evans, 2011, p. 5). Así, no podemos sustraer los productos
audiovisuales de su relación con el contexto histórico y es necesario
observarlos como generadores de cultura. Por ejemplo, Netflix, que cuenta con
un amplio número de suscriptores, ofrece productos que pertenecen a canales
de televisión y se trasmiten a través de la plataforma; pensemos en The Walking
Dead, How to get away with murder, Down Town Abbey, por mencionar
algunos ejemplos. Estos programas y los dispositivos que los hacen posibles
han permitido el surgimiento de relatos interrelacionados que obtienen el
nombre de narrativas transmedia.
La “transmedialidad” (transmediality) y los movimientos relacionados con los
contenidos audiovisuales que cruzan las diversas tecnologías (Evans, 2011, p.
7) es un elemento fundamental para la permanencia del medio. Si los múltiples
tipos de contenido están disponibles a través del mismo medio, un televidente
que comprendía la definición tradicional de la televisión tiene que enfrentarse
a una nueva definición sin lo que se conocía como el broadcasting. La
distribución y exhibición han cambiado, así como los procesos de producción
y los procesos creativos.
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Es importante comprender la diferencia que existe entre las expectativas de la
tecnología y lo que significa el transmedia, término que se relaciona con la
estructura del texto: la televisión es un medio que permite la exploración
narrativa a través de diversas plataformas. Es decir, el transmedia permite
extender el relato a través de otros espacios que van más allá de lo narrado en
el producto original. Considero que observar las cualidades de la televisión y
del streaming como generadores de cultura es un elemento esencial que nos
permite comprender la cultura en la sociedad contemporánea.
Carlos Scolari anota que la narrativa transmedia puede definirse como un tipo
de relato en el que la historia se despliega en múltiples medios y plataformas
de comunicación en el que los consumidores asumen un rol activo en este
proceso (2013, p. 40). Las narrativas audiovisuales de los últimos años,
principalmente a partir del surgimiento de las plataformas streaming, parecen
contar con esta cualidad de manera irremediable; los contenidos que se ofrecen
al espectador cuentan con una amplia variedad: trailers, páginas de internet
(algunas con el antenombre wiki, como Wikigameofthrones), reportajes que
discuten contenidos en diversas plataformas, blogs, entre otros. Incluso, se
crean narrativas interactivas con los videojuegos. Un buen ejemplo de ello es
la serie de ciencia ficción inglesa de Netflix Black Mirror, creada en 2011 por
Charlie Brooker, y específicamente la película Bandersnatch, dirigida por
David Slade y estrenada en 2018. Esta permite al espectador tomar un papel
participativo en el desarrollo de la historia debido a que puede contribuir en las
decisiones sobre las acciones que realiza el protagonista del relato, tal como
sucede con la interacción con el videojuego; la condición del protagonista
como un desarrollador del mismo juego establece una serie de metarrelatos que
determinan el giro de la historia, tanto para el personaje como para el
espectador, en donde la toma de decisiones es la que ordena la trama y el
desenlace positivo o negativo en la acción.
Black Mirror es una serie en la cual lo audiovisual y la tecnología son temas
fundamentales; pese a esto, no existe continuidad, cada episodio expone una
historia con su inicio, desarrollo y desenlace, con un leitmotiv coincidente en
cada uno: los personajes se enfrentan al impacto del uso de la tecnología y de
las narrativas audiovisuales resultado de la misma; estos establecen un vínculo
estrecho con estos dos elementos, mismo que los obligan a modificar su
entorno, generalmente de manera negativa.
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Imagen 4. Escena de Black Mirror: Bandersnatch
Bandersnatch cuenta con la temporalidad de un largometraje. No obstante, a
diferencia de lo que sucede con el resto de los capítulos de la serie, en este se
observa y se vive el impacto de los medios de comunicación en el contexto
futurista en el que tiene lugar la diégesis, ya que el espectador participa de la
toma de decisión de manera interactiva; el receptor es quien puede llevar la
historia a un desenlace fatídico para los personajes. De hecho, pareciera que
las posibilidades de un final afortunado no son posibles, de tal manera que el
mundo de dependencia y fatalidad que condicionan nuestra relación con la
comunicación audiovisual impacta la realidad cotidiana del espectador de
Black Mirror a través de Bandersnatch, de forma tal que el público se va
encaminando a la realidad misma que se presenta en la serie.
La serie hace una fuerte crítica a nuestra relación con la narrativa de los medios
y con la tecnología, al punto que pone de manifiesto lo expresado por diversos
teóricos de la comunicación. Recordemos que Gilles Lipovesky anunció la
fatalidad de la hipermodernidad, a partir del impacto que tienen el consumo a
través de los medios de comunicación en la sociedad, no solo desde una
perspectiva cultural, sino también política y de acción social:
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Himno a las vacaciones, entertainment televisivo, «telemasacre», política
espectáculo y espectáculo publicitario: allí donde se sacralizaba la abnegación,
tenemos ahora la evasión y la violencia en zoom; donde se santificaba la pureza de
intenciones, tenemos los escalofríos de la violencia mediática y la frivolidad de las
cosas; allí donde se beatificaba la grandeza de superarse, tenemos el erotismo en
autoservicio, las comodidades del confort, el poder disuasivo de la publicidad. En
nuestras sociedades, los objetos y marcas se exhiben más que las exhortaciones
morales, los requerimientos materiales predominan sobre la obligación
humanitaria, las necesidades sobre la virtud, el bienestar sobre el Bien. La era
moralista tenía como ambición la disciplina del deseo, nosotros lo exacerbamos;
exhortaba a los deberes hacia uno mismo y hacia los demás, nosotros invitamos a
la comodidad (2000, pp. 52-53).
Los productos audiovisuales forman parte de la vida cotidiana. Ya sea que se
trate de contenidos streaming, contenidos en redes sociales, plataformas o a
través de la televisión tradicional, los espectadores se insertan en una dinámica
de interacción social y cultural. En este sentido, la transmedialidad juega un
papel central en la construcción del mundo ficcional en términos de lo que
llamamos la hiperdiégesis, la creación de un vasto y detallado mundo narrativo
espacial, y solo una fracción de ese espacio es directamente visto o se encuentra
con el texto original (Evans, 2011, p. 18). Los relatos transmedia proveen
partes del texto que no están disponibles a través de los episodios televisivos;
además, ofrecen una visión ficcional que complementa la televisiva y que
permite múltiples mundos de acceso al mundo ficcional. El transmedia implica
la creación de un sistema de intercambio cultural –incluso artístico– que tiene
un impacto en nuestro contexto histórico.
Es importante reflexionar sobre las formas en las que nos relacionamos con los
medios de comunicación y, específicamente, con los contenidos audiovisuales
debido a que estos permean todos los aspectos de nuestra vida cotidiana.
Anteriormente se cumplía, de manera cabal, la idea de Umberto Eco: “una
determinada relación con el público, conducida a través de un medio dado,
constituye calificar una expresión incluso en sus componentes estéticos”
(2006, p. 314). En la actualidad, la actitud receptiva es diferente y, si bien hace
algunos años se encontraba determinada por el medio, en nuestros días se
encamina a estar determinada por la forma, independientemente del medio;
como señala Mattelart, todo está condicionado por la ley de la oferta y la
demanda. Además, como apunta Lipovetsky, “el conflicto estructural del
individualismo se juega el porvenir de las democracias: no hay en absoluto
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tarea más crucial que hacer retroceder el individualismo irresponsable,
redefinir las condiciones políticas, sociales, empresariales, escolares, capaces
de hacer progresar el individualismo responsable” (2000, p. 16). Sin embargo,
pareciera que las políticas en torno a la producción, consumo y
comercialización de lo audiovisual se inclinan cada vez más por la
individualización sin control, principalmente en los países en vías de
desarrollo.
Probablemente, como señala Carlos Scolari en su texto Hipermedicaciones,
es necesario reflexionar en las teorías de la comunicación de masas como las que
hablan de los medios masivos y de la dinámica que oponen las estrategias de
manipulación a los procesos de interpretación. La novedad de lo digital llega por lo
tanto a un campo fluctuante, aportando aún más inestabilidad a las conversaciones
sobre los medios (2008, p. 33).
Los seres humanos hemos condicionado las formas de nuestras relaciones con
los productos audiovisuales en virtud del uso de la tecnología. No obstante, las
teorías de la comunicación tienen su sustento en una serie de procesos
históricos que fueron fundamentales para la humanidad y que no podemos
perder de vista al analizar los discursos audiovisuales contemporáneos.
FUENTES DE CONSULTA
ALBÈRA, F. (comp.) (1998). Los formalistas rusos y el cine. La poética del
filme. Trad. J. Á. Alcalde. Barcelona, España: Paidós.
ECO, U. (2006). Apocalípticos e integrados. Trad. A. Boglar. Ciudad de
México, México: Tusquets.
EVANS, E. (2011). Transmedia television. Audiences, new media and daily
life,
Nueva
York,
U.
S.
A.:
Routledge.
https://doi.org/10.4324/9780203819104
LIPOVETSKY, G. (2000). El crepúsculo del deber. La ética indolora de los
nuevos tiempos democráticos. Barcelona, España: Anagrama.
MATTELART, A. (2002). Geopolítica de la cultura, Bogotá, Colombia:
Desde abajo.
MATTELART, A. (1996). La mundialización en la comunicación, Barcelona,
España: Paidós.
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MATTELART, A. y Mattelart, M. (2003). Historia de las teorías de la
comunicación, Buenos Aires, Argentina: Paidós.
RUSSELL, M. (productor), Salde D. (director) (2018). Black Mirror:
Bandersnatch [película cinematográfica], Reino Unido: Netflix.
SCOLARI, C. (2008). Hipermediaciones. Elementos para una teoría de la
comunicación digital interactiva. Barcelona, España: Gedisa.
SCOLARI, C. (2013). Narrativas transmedia: cuando todos los medios
cuentan, Barcelona, España: Deusto.
WILLIAMS, R. (1992). Historia de la comunicación. Del lenguaje a la
escritura. V. 1. Barcelona, España: Bosch Comunicación.
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REVISTA DE ESTUDIOS E INVESTIGACIÓN
EN PSICOLOGÍA Y EDUCACIÓN
eISSN: 2386-7418, 2015, Vol. Extr., No. 11.
DOI: 10.17979/reipe.2015.0.11.639
Teoría del Apego y Síndrome de Down: conceptos básicos y claves educativas
Yanina Guerrero y Eduardo Barca-Enríquez
Universidade da Coruña
Resumen
El nacimiento de un hijo generalmente predispone
a los padres para desarrollar una crianza adecuada.
Sin embargo, cuando el bebé expresa una alteración
en su desarrollo que “dista de la evolución
esperada”, se rompen las expectativas habituales y el
proceso de vinculación afectiva puede verse
mermado. Se altera así el desarrollo de un apego
seguro. Pretendemos recopilar y organizar toda la
información bibliográfica al respecto, resaltando las
principales claves para favorecer un apego seguro en
niños con Síndrome de Down, permitiendo un mejor
desarrollo integral con el objetivo de posibilitar el
logro de los mayores potenciales de cada uno.
Palabras clave: Síndrome de Down, teoría del
apego, situación del extraño, claves educativas
En lo que se refiere al ámbito de la intervención de
corte psicosocial y clínica, es más que corriente recurrir a
la Teoría del Apego para basar la explicación de la
relación entre un menor y sus cuidadores. Dicha teoría
tiene sus raíces en el psicoanálisis, en la etiología y en las
teorías cognitivas-conductuales, y fue organizada y
sistematizada por Bowlby. Este considera que el niño
viene al mundo preparado biológicamente para
establecer un estrecho lazo de unión con su figura de
cuidado. Este autor formula los supuestos básicos de su
teoría sobre la motivación y la regulación de la conducta.
Considera que existen diversos sistemas conductuales
que están diseñados para alcanzar una meta concreta. De
esta forma, la función del sistema conductual del apego
es la supervivencia del individuo y de la especie en
general.
Las aportaciones de Mary Main y colaboradores, junto
con los estudios de Ainsworth, ayudan a la expansión de
dicha teoría añadiendo un estilo más y dando así mayor
consolidación a lo que parecía ser una interesante y
enriquecedora línea de trabajo. Ainsworth (1983), define
el apego como aquellas conductas que favorecen ante
todo la cercanía con una persona determinada. Es mutuo
y recíproco. Diseña una de las técnicas más fructíferas en
el estudio del apego como es la Situación Extraña, la cual
contribuye a que Mary Main descubra una nueva
categoría de estilos de apego. Así Bowlby, Ainsworth y
Main serán las figuras con mayor peso en el tema del
apego, y quienes mayores contribuciones han hecho a
este ámbito.
El apego en niños con Síndrome de Down
El nacimiento de un hijo suele ser un acontecimiento
que genera satisfacción y alegría. Sin embargo, cuando el
bebé expresa una diferencia visible y notoria que “se
aleja de la normalidad”, se rompen las expectativas y el
proceso de vinculación afectiva puede verse modificado
o mermado. Esta situación se hace más pronunciada
cuanto más visibles sean las discapacidades que presente
el bebé, como ocurre en los casos de Síndrome de Down.
Para evitar casos de maltrato, abandono o
discriminación, concretamente en el caso de niños con
Síndrome de Down, es necesario que el equipo de salud
perinatal elabore anticipadamente estrategias que
faciliten y refuercen en el momento en el que se
diagnostica al pequeño, el apego y la vinculación
afectiva, principalmente después del parto. En caso de
niños autistas, deberá tenerse en cuenta todas estas
directrices en el momento de su diagnóstico y
comunicación a sus progenitores. Se considera
fundamental y primordial cuidar el lenguaje que se
emplea, la información que se brinda y respetar las etapas
emocionales por las cuales atraviesan los padres. Es
importante atender estos aspectos puesto que la actitud
de sus padres influirá con casi absoluta seguridad en su
capacidad de integración social y emocional del hijo a la
familia y a la sociedad.
Como sabemos, el proceso de vinculación afectiva es
algo conocido y su origen es ancestral. Se ha
determinado además que esta conducta es predecible y
evolutiva, esto es, que se produce por etapas
secuenciadas (Rossel, 2004).
Etapas de la vinculación afectiva
• Primera etapa: corresponde al periodo del
embarazo. Durante la gestación y los momentos previos
al parto los padres se encuentran ansiosos y cargado de
expectativas positivas con respecto a su hijo. Ese cariño y
afecto que se ha ido formando a lo largo de 9 meses
tomará una forma tangible con el nacimiento del bebé. Se
idealiza al bebé, se piensa y se divaga sobre diversos
aspectos de la vida del futuro hijo. Se dice que se produce
un “enamoramiento” de la imagen del bebé ideal.
• Segunda etapa: comienza inmediatamente después
del parto. En esos momentos, los padres, quienes están
cargado de expectativas positivas sobre su bebé, toman
contacto real y directo con el mismo por primera vez. El
recién nacido, al ser puesto sobre el abdomen de su
madre, posee la capacidad innata de dirigirse hacia el
pezón y succionar firmemente. Este evento que se
produce dentro de la primera media hora de vida, es
extremadamente importante para solidificar la
vinculación afectiva, puesto que es cuando se producen
crecientes descargas de oxitocina a nivel cerebral, se
elevan además lo niveles plasmáticos de opiodes
endógenos, se desencadena un estado de somnolencia y
Correspondencia: Eduardo Barca Enríquez, e.barcae@udc.es
Selección y peer-review bajo responsabilidad del Grupo de Investigación G000422-GIPDAE, Universidade da Coruña, España.
GUERRERO; BARCA-ENRÍQUEZ
ansiedad en la madre con un elevado umbral para el
dolor. Se trata así de un estado emocional difícil de
describir donde la madre, al ver y sentir a su hijo,
completa y consolida la fase de enamoramiento que se
inició en la fase de embarazo, y que marcará la
maternidad de forma positiva
• Tercera etapa: se inicia después del nacimiento. Es
el periodo más largo y evolutivo de los tres, y en el cual
se refuerzan los lazos afectivos que se establecieron en
un primer momento a través del proceso del apego.
Como sabemos, este proceso se caracteriza por la
interacción de ambas partes para que el resultado sea
adecuado y óptimo. Cabe destacar, como se ha
comentado anteriormente, que el niño se va
desarrollando y va creciendo, a la vez que recibe los
estímulos de sus padres y responde de acuerdo a su etapa
de desarrollo.
Factores que alteran el proceso de vincultación y el
desarrollo normal del vínculo de apego
• Los padres: condiciones de salud de los progenitores
desfavorables, principalmente de la madre, que
imposibiliten el contacto inicial. Ejemplo de ello sería
casos de infecciones, cesárea, hipertensión arterial
severa, entre otros. También incluiríamos en este
apartado la salud mental y la estabilidad emocional de la
madre, por ejemplo, deterioro neurológico, madre
adolescente, drogadicción…
• Medio ambiente: hospital con normas rígidas que
entorpecen el contacto precoz de los bebés con sus
padres (visitas restringidas, falta de contacto directo con
el bebé, entre otros); equipo médico poco acogedor y mal
informado (desconoce las redes de apoyo social y los
programas de estimulación neuro-sensorial locales,…);
ausencia de redes de ayuda social de accesibilidad local
(capacidad de integración social,…); medio social y
cultural
intelectualmente
desprotegido
(véase
drogadicción, alcoholismo, prejuicios,…) junto con las
experiencias previas desfavorables.
• Recién nacido: condiciones de salud del bebé que
imposibilitan la permanencia con su madre. Estas
condiciones pueden ser transitorias, prolongadas o
permanentes.
Etapas de transición emocional de los padres con
hijos con discapacidad
Se diferencian diversas etapas emocionales por las
cuales atraviesan los padres cuando tienen un hijo con
discapacidad (Rossel, 2004):
• Primera etapa, impacto: la noticia del diagnóstico
de su bebé los deja paralizados, no logrando comprender
qué está pasando. Sienten que sus expectativas positivas
sobre su hijo se han esfumado y parecen vivir una
pesadilla que dista de la realidad. Se encuentran atónitos,
perplejos. Es posible que rompan a llorar. Unos optan por
preguntar insistentemente por la condición de su hijo,
mientras que otros hacen como si ésta no existiese.
Existen dificultades y están reacios a acoger a su bebé,
principalmente si no han tenido contacto inmediato
posterior al parto puesto que la imagen de su bebé es
“desfavorable” a lo que se esperaban.
Cuanto mayor sea el periodo de distanciamiento
posterior al parto, mayores serán dificultades tendrán
para enfrentar a su hijo
• Segunda etapa, negación: posteriormente, los
padres atraviesan una etapa de negación de la realidad.
Se resisten a aceptar la situación de su hijo. Es posible
que hasta lleguen a verbalizar “que éste no es su hijo” o
“debe de ser un error”, “no tiene lo que dicen”, “están
equivocados”, entre otros. Los prejuicios y las opiniones
de terceros cobran importancia. Existen casos donde los
padres permanecen incomunicados y separados, de
forma que se dificultará aún más la vinculación.
• Tercera etapa, tristeza o dolor: los padres ya toman
conciencia de la realidad e intentan comprender en parte
lo que les ocurre. Se va consolidando paulatinamente la
imagen de pérdida del bebé ideal, de los sueños
esfumados y perdidos y de la esperanza ya inexistente.
Todo ello conllevará un profundo sentimiento de dolor,
tristeza y angustia. Pueden expresarlo mediante
impotencia, frustración y posiblemente sentimientos de
culpabilidad. Se encuentran realmente angustiados, le
temen a lo desconocido.. Sufren cuando perciben la
dificultad de su vinculación, aunque son más próximas
que en las etapas anteriores y acceden a tomar a sus hijos,
amamantarlos y expresan sentimientos de ternura y dolor
a la vez.
La duración de esta etapa es variable y depende en gran
parte de cómo han sido resueltas las etapas anteriores.
Los padres buscarán en sus hijos señales de interacción
(succión energética al pecho, una sonrisa, fijación de la
mirada,…). Sin embargo, es posible que no todos los
bebés que padecen Síndrome de Down pueden responder
a estas demandas al nacer debido a su hipotonía. Ello
conlleva frustración de los padres y los desalienta
enormemente, llegando incluso a creer que su hijo “no
los necesita”. Esta ansiedad, junto al dolor que
experimentan, propicia a que ciertos padres pierdan las
esperanzas de vincularse con su bebé.
• Cuarta etapa, adaptación: entran en un proceso de
adaptación que se caracteriza por aceptar la condición de
su hijo y los sentimientos que están experimentando. Se
muestran interesados en aprender y conocer más sobre el
Síndrome de Down. Es de suma importancia la
aceptación que el medio familiar ha expresado a este
nuevo integrante así como el apoyo que la pareja exprese
al otro miembro. El peso del ambiente, en caso de ser
nocivo y negativo, puede exponer al niño a riesgos de
desvinculación afectiva, abandono o maltrato.
• Quinta etapa, reorganización: el bebé ha sido
completamente integrado a la familia, la cual ha
aprendido a conocer y tolerar sus discapacidades y se
comprometen e implican en su rehabilitación y en su
adaptación al medio. La vinculación afectiva se
construye a la par que se fortalece cada día mediante los
avances emocionales y neurológicos del bebé, facilitando
así la interacción con sus padres, convirtiendo así este
proceso en una retroalimentación positiva. Los
programas de estimulación temprana facilitarán la
consecución de esta retroalimentación, en la cual los
padres tienen la oportunidad de relacionarse con un
R Est Inv Psico y Educ, 2015, Extr.(11), A11-099
TEORÍA DEL APEGO Y SÍNDROME DE DOWN
equipo multidiscilplinar y con otros padres en situaciones
similares a la suya.
Cabe destacar que la temporalidad con la que se
producen las anteriores etapas no está muy bien definida.
La duración de cada una de ellas varía y dependerá
principalmente de la influencia del medio ambiente sobre
los progenitores y los rasgos emocionales de los mismos.
Es posible que los padres permanezcan más tiempo o
incluso se queden estancados emocionalmente en alguna
de las etapas iniciales (en las tres primeras),
impidiéndoles avanzar hacia las etapas de resolución y
adaptación.
Investigación sobre Apego y Síndrome de Down
A su vez, diversos autores han abordado y estudiado el
papel del apego en niños con Síndrome de Down. En
concreto, Beeghly, Weiss-Perry, & Cicchetti (1990)
consideraban que la construcción por parte de estos niños
de unas relaciones seguras de apego entre los doce y los
veinticuatro meses era la consecuencia de la interacción
con sus cuidadores, siendo así el apego seguro el patrón
normativo de los niños con esta discapacidad. Sin
embargo, se crean ciertas dudas sobre la validez de la
Situación Extraña como medio de evaluación del apego
en estos niños puesto que muchos de ellos no parecen
denotar estrés durante los episodios de separación y
reunión (Vaughn y Waters, 1990), aunque los pequeños
son conscientes de la separación.
Otros profesionales como Main y Solomon (1990)
indicaron que no es conveniente utilizar la categoría de
apego desorganizado cuando se investigan a niños con
discapacidades físicas y/o mentales ya que muchos de
ellos pueden presentar elementos de desorganización
conductual. Esta desorganización puede ser resultado
más de su discapacidad que del fracaso en la
construcción de una estrategia coherente de organización
de la conducta de apego.
Vaughn, Goldberg, Atkinson, Marcovitch, MacGregor
y Seifer (1994) estudiaron tres muestras independientes
con Síndrome de Down de entre doce y veinticuatro
meses de edad. Se empleó para su evaluación la
Situación Extraña. Los resultados que se obtuvieron
fueron los siguientes: el 42% de los niños no pudo ser
clasificado en un tipo de apego concreto utilizando el
procedimiento tradicional, siendo categorizados como
inseguros/inclasificables.
Además,
se
muestran
diferencias significativas entre ambos grupos en
seguridad del apego. Así, era más probable que los
pertenecientes al grupo de comparación presentaran un
apego seguro, aunque la tasa superior de inseguros entre
los niños con Síndrome de Down podría atribuirse a la
imposibilidad de clasificar a muchos de ellos. Incluso
cuando se descartan estos casos inclasificables de los
análisis, el porcentaje en el grupo de comparación fue
mayor que entre los del Síndrome de Down (38% frente a
un 21%). Se encuentran diferencias también en la
distribución de los tres tipos de apego. Esto puede
deberse fundamentalmente a la baja proporción de
ambivalentes entre los niños con este síndrome. Se
observan diferencias en las escalas de interacción, esto
es, en la búsqueda de proximidad, mantenimiento del
contacto, resistencia al contacto/interacción y en el
número de intervalo de llanto. Los niños que padecían
Síndrome de Down puntuaron menos en estas escalas,
excepto en la conducta de evitación.
Como se ve, son unos resultados poco esclarecedores y
su interpretación resulta algo compleja. Por un lado,
podemos pensar que las diferencias observadas son
superficiales y que con una selección adecuada de sujetos
se encontrarían unos patrones de apego similares. Los
niños con Síndrome de Down se apegan a sus madres y la
mayoría de ellos establece unas relaciones seguras de
apego con sus progenitoras.
Vaughn et al. (1994) interpretaron los resultados de un
modo totalmente diferente al que hemos explicado líneas
arriba. Ellos consideran que la Situación Extraña provoca
en los niños con este síndrome unos niveles de estrés
funcionalmente distintos. Defienden que ellos raramente
exteriorizaban su estrés (ni siquiera cuando se quedaban
solos). Este nivel bajo de activación/estrés propiciaba
que no buscaran el contacto durante los reencuentros,
aunque se aproximaran e interactuaran. Además, la falta
de contacto y de necesidad de que los tranquilizaran
hacían que difícilmente presentaran una conducta de
resistencia.
Así, se considera que el diferente impacto psicológico
de los episodios de separación y reunión pone de
manifiesto la incapacidad del procedimiento de
Ainsworth (seguro, evitativo, preocupado) y de Main y
Solomon (desorganizado) para la captación de las
capacidades y diferencias y no evalúen el patrón de
apego de estos niños. De esta forma, este procedimiento
no debería utilizarse ni para evaluar ni para valorar de
forma rutinaria la conducta en los niños con Síndrome de
Down en la Situación Extraña.
También puede ocurrir que los procesos necesarios
para la consolidación del apego requieran más tiempo en
el caso de estos niños. Debemos destacar además dos
estudios de niños con Síndrome de Down en los que la
calidad de las relaciones de apego se evaluó con el
Attachment Q-set (AQS; Waters y Deane, 1985) y con la
Situación Extraña. Vaughn, Leftover, Seifer & Barglow
(1989) demuestran que los tipos de apego según la
Situación Extraña no se relacionaban con las
puntuaciones en seguridad del apego con las obtenidas en
el AQS. Esto podría ser una prueba de que las
clasificaciones definidas en la Situación Extraña no
constituyen un índice válido de la seguridad del apego en
estos niños.
Sin embargo, en otro estudio, Atkinson y sus
colaboradores encontraron una relación significativa
entre el tipo de apego obtenido en la Situación Extraña y
la puntuación obtenida en seguridad en el AQS cuando
los niños tenían veintidós meses de edad. Por lo tanto,
cabe la posibilidad que los procesos de consolidación del
apego en niños con Síndrome de Down lleven más
tiempo. Atkinson et al. (1999) evaluaron la influencia del
funcionamiento adaptativo/intelectual del niño y de la
sensibilidad de la madre, y la interacción entre ambas
variables, sobre la seguridad del apego.
El estudio consistía en medir cuatro constructos en dos
ocasiones diferentes durante dos años. Estos cuatro
constructos serían los siguientes: funcionamiento
cognitivo del niño, seguridad del apego, sensibilidad
R Est Inv Psico y Educ, 2015, Extr.(11), A11-100
GUERRERO; BARCA-ENRÍQUEZ
materna y funcionamiento adaptativo. La muestra contó
con 53 niños de entre catorce y treinta meses de edad
cronológica en la primera observación y sus madres.
Siete meses después, al pasar por la situación extraña por
primera vez, los niños tenían una edad mental de entre
doce y veintitrés meses según el Bayley. La conducta de
la Situación Extraña se codificó en los tipos de apego A,
B, C, D y U (aquellos que no pudieron ser clasificados en
ninguna categoría). Estos últimos se caracterizaban por
una aparente despreocupación aun dándose cuenta de que
su madre abandonaba la habitación, o incluso si la
separación les inquietaba, se tranquilizaban con la
presencia de un extraño.
También se observaron las relaciones de la madre y el
hijo en el hogar en dos ocasiones cubriéndose el AQS. La
sensibilidad de la madre se evaluó en seis ocasiones
durante dos años, en diferentes lugares (laboratorio y
hogar), diferentes situaciones (mientras contestaba a los
cuestionarios, mientras lo alimentaba, mientras se
entrevistaba a la madre) y con diversos procedimientos.
Se valoran cuatro constructos generales basados en las
escalas de Ainsworth: accesibilidad (disponibilidad
psicológicamente cuando la necesita el hijo), sensibilidad
(respuesta
apropiada
y
rápida),
cooperación
(intervención mínimamente intrusiva) y aceptación (la
madre valora la voluntad del niño). A mayores se utilizó
el Maternal Behavior Q-set para evaluar la respuesta de
las madres a las señales del hijo cuando tenía cuarenta y
dos meses.
Los resultados de este estudio fueron los siguientes: a
los veintitrés meses un 40% mostraba un apego seguro,
47% no se puedo clasificar en ninguna categoría y un
13% mostraba claros indicio de apego inseguro.
Dieciséis meses después se obtuvo una distribución
similar (47,5%, 32,5% y 20% respectivamente). El 62%
fue clasificado en el mismo tipo de apego según la
Situación Extraña a los veintiséis y a los cuarenta y dos
meses según el AQS. Además, la mayoría no había
desarrollado una conducta de base segura incluso en
preescolar. Las diversas medidas de sensibilidad materna
se relacionaban con los tipos de apego en la Situación
Extraña y con la seguridad del apego (AQS) evaluada a
los cuarenta y dos meses de edad. Las madres de los
niños con apego seguro eran más sensibles que las de los
inseguros y que las de los inclasificables.
Por otro lado, el funcionamiento mental predecía la
conducta de los niños en la Situación Extraña y la
seguridad del apego según el AQS. De esta forma, los
que fueran más competentes cognitivamente era más
probable que formaran un apego seguro con sus madres.
Los análisis de regresión demostraron que la interacción
entre sensibilidad de la madre y el funcionamiento
cognitivo del niño predecía la seguridad del apego. Así,
la obtención de puntuaciones elevadas en las dos
variables incrementaba la probabilidad de que se hubiera
desarrollado apego seguro. Además, se constata que la
conducta de apego mostrada en el laboratorio era un
reflejo bastante exacto de la conducta de base segura
mostrada en el hogar.
Podríamos concluir pues, en cuanto a los estudios y la
determinación de los estilos de apego en niños con
Síndrome de Down, el planteamiento de ciertas dudas
sobre la validez de la Situación Extraña en esta
población, puesto que muchos de estos niños no parecen
experimentar estrés durante estos episodios de
separación aunque sean conscientes de ello. Además, la
categoría de apego desorganizado no podría utilizarse en
estos niños por los motivos ya explicados anteriormente,
que se referían que dicha desorganización podría deberse
más a la discapacidad que a la falta de una estrategia
coherente de organización de la conducta de apego. Se ha
constatado que hasta un 42% de niños con Síndrome de
Down no pueden ser clasificados y que puntúan meno en
la búsqueda de proximidad, resistencia al
contacto/interacción y al mantenimiento del contacto.
Es posible que ocurra también que los procesos
necesarios para que se consolide el apego necesiten de
más tiempo en caso de niños con esta discapacidad.
Atkinson et al. (1999) encontraron que la mayoría no
había desarrollado una conducta de base segura incluso
en preescolar, ni en caso de tratarse de una circunstancia
rutinaria en las que el apego y la exploración estaban
libres de estrés (AQS) ni en el caso de dar respuesta a una
emergencia (Situación Extraña). La conducta de apego
que se muestra en el laboratorio era un reflejo bastante
fidedigno de la conducta de base segura mostrada en el
hogar del niño. El 47% no pudo ser clasificado a los dos
años, mientras que un 32% no puedo ser clasificado
tampoco a los cuatro años. Se constata también que la
interacción entre sensibilidad materna y el
funcionamiento cognitivo del hijo predecía la seguridad
del apego.
Claves educativas generales
Con todos los conocimientos y demostraciones de
evidencias claras hasta aquí señaladas, consideramos
pues que el apego también es una emergencia afectiva en
los niños con Síndrome de Down. Por ello, debemos
tener en consideración la importancia de un cambio en
los modelos afectivos de los padres de dichos niños. Así,
es conveniente no sólo comprender cada una de las
etapas por las cuales atraviesan los progenitores descritas
anteriormente, sino también tenerlas en consideración y
respetarlas. No debemos interferir con el curso natural de
los eventos de la vinculación, sino que tenemos que hacer
todo lo posible por facilitar el contacto físico inmediato
posterior al parto entre los padres y los bebés,
principalmente con la madre. Simplemente con colocarlo
sobre el pecho de la misma, ya estamos iniciando el
apego. Todo ello es conveniente de hacerlo antes de
brindarle y comunicarles toda la información al respecto
de su hijo. Es preferible hacerlo una vez hayan tomado
contacto físico con su bebé. Debemos hacer hincapié en
que se concentren en las necesidades afectivas, en la
alimentación de su bebé, en el calor que le deben brindar
a éste, y que la información al respecto de la
discapacidad de su hijo se le brindará más adelante.
Es importante señalar que si por diversas razones el
bebé debe permanecer separado de sus padres, es
recomendable intentar reducir al máximo este tiempo de
distanciamiento. Diversos autores señalan que más de 24
horas separados dificultaría el restablecimiento del apego
R Est Inv Psico y Educ, 2015, Extr.(11), A11-101
TEORÍA DEL APEGO Y SÍNDROME DE DOWN
puesto que los padres se encuentran en la etapa de
impacto. Es conveniente no presionarlos y limitarse a
preguntar si desean ver y conocer a su bebé, darle el
pecho o cogerlo en brazos. Si estos responden
negativamente a nuestra cuestión, no hay que forzarlos
para que lo hagan, ya que estarían en la etapa de
negación.
Cuando esta etapa finalice, los padres accederán a
coger a su bebé, con miedo pero sin rechazo. Sabemos
que éstos necesitan tiempo para superar este duro
momento. Si se encuentran en la etapa de negación o de
dolor no es conveniente darles demasiados detalles
médicos, sino que es más beneficioso brindarles apoyo y
preguntarles cómo se siente, propiciando así la expresión
de sus emocionales, sentimientos, temores, prejuicios o
creencias acerca de la situación por la que pasan,
llegando incluso a llorar. En ese momento no debemos ni
educarlos, ni hablarles de la condición de salud de su
bebé ni parecido, sino arroparlos, acogerlos,
familiarizarlos con los sentimientos, con la expresión de
los mismos, apoyarlos y animarlos en esta dura
circunstancia. Deben encontrarse consigo mismos,
enfrentarse a su dolor, ser conscientes de ello y
manifestarlo sin pudor ni culpabilidad.
Una vez que los padres alcanzan la etapa de
adaptación, comenzarán a preguntarse qué van a hacer en
un futuro, y empieza a visualizarse el lazo afectivo hacia
su hijo. Es ahí cuando debemos reforzar esta vinculación,
mostrándoles señales de interacción que su hijo está
logrando y educarlo sobre la vigilancia en la salud y
sobre los programas de estimulación temprana, el rol de
los padres de apoyo, así como la protección legal y social
que ofrece el Estado.
Es por ello que el apego, especialmente en situaciones
de discapacidad del bebé, se transforma en una
emergencia afectiva, ya que repercutirá en el futuro del
bebé. El equipo sanitario debe atender a las etapas por las
que pasan los padres para detectar alguna anomalía (por
ejemplo, duración prolongada o estancamiento en alguna
de ellas) que pueda repercutir en el pequeño y la cual
requiera la intervención de un profesional que le asista
ante esa situación.
También es necesario estar alerta de las situaciones
ambientales que contribuyan o favorezcan el deterioro de
la vinculación afectiva, puesto que pueden entorpecer el
desarrollo normal del niño. La desvinculación afectiva en
el Síndrome de Down existe y es una realidad que
debemos tener en cuenta. Por ello, es conveniente
elaborar estrategias para evitarla o minimizarla en caso
de que ya esté presente. Así, el equipo de salud debe
informarse y establecer un plan de trabajo que consista
en asistir emocionalmente a estas familias de la forma
correcta y especializada.
Referencias bibliográficas
Ainsworth, M. D. S. (1983). Models of achievement:
Reflections of prominent women in psychology. Nueva
York:Columbia University Press.Atkinson, P., Coffey,
A., & Delamont, S. (1999). Ethnography: Post, Past and
Present. Journal of Contemporary Ethnography, 28(5),
460–71.
Beeghly, M., Weiss-Perry, B., & Cicchetti, D. (1990).
Beyond sensorimotor functioning: Early communicative
and play development of children with Down syndrome.
In D. Cicchetti & M. Beeghly (Eds.), Children with
Down syndrome: A developmental perspective (pp.
329-368). New York: Cambridge University Press.
Main, M., & Solomon, J. (1990). Procedures for dentifying
disorganized/disoriented infants during the Ainsworth
Strange Situation. In M. Greenberg, D. Cicchetti & M.
Cummings (Eds), Attachment in the preschool years (pp.
121-160). Chicago: University of Chicago Press.
Rossel, K. (2004). Apego y vinculación en el Síndrome de
Down. Una emergencia afectiva. Revista Pediatría
Electrónica, 1(1), 3-8. Recuperado de:
http://www.portalsindromededown.com.br/arquivos/ape
go_e_down.pdf
Vaughn, B. E., & Waters, E. (1990). Attachment behavior
at home and in the laboratory: Q-sort observations and
strange situation classifications of one-year-olds. Child
Development, 61, 1965 – 1973.
Vaughn, B. E., Leftover, G. B., Seifer, R. & Barglow, P.
(1989). Attachment behavior, attachment and
temperament during infancy. Child Development,
60,728-37.
Vaughn, B. E., Goldberg, S., Atkinson, L., Marcovitch,
S., MacGregor, D., & Seifer, R. (1994). Quality of
toddler-mother attachment in children with Down
syndrome: Limits to interpretation of strange situation
behavior. Child Development, 65, 95-108.
R Est Inv Psico y Educ, 2015, Extr.(11), A11-102.
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The Contributions of HIF-Target Genes to Tumor Growth in RCC
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Abstract * E-mail: haifeng.yang@jefferson.edu
¤ Current address: Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, Pennsylvania, United States of
America Citation: Zhang T, Niu X, Liao L, Cho E-A, Yang H (2013) The Contributions of HIF-Target Genes to Tumor Growth in RCC. PLoS ONE 8(11): e80544. doi:10.1371/journal.pone.0080544 X, Liao L, Cho E-A, Yang H (2013) The Contributions of HIF-Target Genes to Tumor Growth in RCC. PLoS ONE 8(11): e80544. ation: Zhang T, Niu X, Liao L, Cho E-A, Yang H (2013) The Contributions of HIF-Target Genes to Tumor Growth in RCC. PLoS
:10.1371/journal.pone.0080544 Received August 15, 2013; Accepted October 14, 2013; Published November 18, 2013 Zhang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which perm
stribution, and reproduction in any medium, provided the original author and source are credited. Funding: This study was funded in part by National Institute of Health (CA155015 to HY), VHL Family Alliance (to HY) and Chinese Scholar Council (to
TZ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. ¤ Current address: Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, Pennsylvania, United States of
America ¤ Current address: Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, Pennsylvania, United States of
America ¤ Current address: Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, Pennsylva
America Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, Pennsylvania, United States of Ting Zhang1, Xiaohua Niu2, Lili Liao2¤, Eun-Ah Cho3, Haifeng Yang2*¤ 1 Tianjin Institute of Urology, Second Hospital of Tianjin Medical University, Tianjin, China, 2 Department of Cancer Biology, Lerner Research Institute,
Cleveland Clinic, Cleveland, Ohio, United States of America, 3 Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia,
Pennsylvania, United States of America The Contributions of HIF-Target Genes to Tumor Growth
in RCC Ting Zhang1, Xiaohua Niu2, Lili Liao2¤, Eun-Ah Cho3, Haifeng Yang2*¤ Abstract Somatic mutations or loss of expression of tumor suppressor VHL happen in the vast majority of clear cell Renal Cell
Carcinoma, and it’s causal for kidney cancer development. Without VHL, constitutively active transcription factor HIF
is strongly oncogenic and is essential for tumor growth. However, the contribution of individual HIF-responsive genes
to tumor growth is not well understood. In this study we examined the contribution of important HIF-responsive genes
such as VEGF, CCND1, ANGPTL4, EGLN3, ENO2, GLUT1 and IGFBP3 to tumor growth in a xenograft model using
immune-compromised nude mice. We found that the suppression of VEGF or CCND1 impaired tumor growth,
suggesting that they are tumor-promoting genes. We further discovered that the lack of ANGPTL4, EGLN3 or ENO2
expression did not change tumor growth. Surprisingly, depletion of GLUT1 or IGFBP3 significantly increased tumor
growth, suggesting that they have tumor-inhibitory functions. Depletion of IGFBP3 did not lead to obvious activation
of IGFIR. Unexpectedly, the depletion of IGFIR protein led to significant increase of IGFBP3 at both the protein and
mRNA levels. Concomitantly, the tumor growth was greatly impaired, suggesting that IGFBP3 might suppress tumor
growth in an IGFIR-independent manner. In summary, although the overall transcriptional activity of HIF is strongly
tumor-promoting, the expression of each individual HIF-responsive gene could either enhance, reduce or do nothing
to the kidney cancer tumor growth. Citation: Zhang T, Niu X, Liao L, Cho E-A, Yang H (2013) The Contributions of HIF-Target Genes to Tumor Growth in RCC. PLoS ONE 8(11): e80544. doi:10.1371/journal.pone.0080544
Editor: Peiwen Fei, University of Hawaii Cancer Center, United States of America
Received August 15, 2013; Accepted October 14, 2013; Published November 18, 2013
Copyright: © 2013 Zhang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This study was funded in part by National Institute of Health (CA155015 to HY), VHL Family Alliance (to HY) and Chinese Scholar Council (to
TZ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing interests: The authors have declared that no competing interests exist. Table 1. Table 1. Name of shRNA constructs
shRNA sequences
HIF1b-1770
GAGAAGTCAGATGGTTTATTT
HIF2a-1631
CGACCTGAAGATTGAAGTGAT
HIF2a-566
CCATGAGGAGATTCGTGAGAA
VEGF-1137
AGGGCAGAATCATCACGAAGT
VEGF-1500
GCGCAAGAAATCCCGGTATAA
VEGF-1587
GACGTGTAAATGTTCCTGCAA
CCND1-326
GCCCTCGGTGTCCTACTTCAA
CCND1-539
CTCTAAGATGAAGGAGACCAT
CCND1-2322
GCCAGGATGATAAGTTCCTTT
ANGPTL4-786
GCAGAGTGGACTATTTGAAAT
ANGPTL4-1311
GAAGCTTAAGAAGGGAATCTT
EGLN3-692
GTGGCTTGCTATCCGGGAAAT
ENO2-1992
CGCCTGGCTAATAAGGCTTTA
ENO2-1735
CGCACTTTCCACTTCTTCCTT
GLUT1-1598
CCAAAGTGATAAGACACCCGA
GLUT1-455
GCGGAATTCAATGCTGATGAT
GLUT1-2310
GCCACACTATTACCATGAGAA
IGFBP3-633
CCTCCATTCAAAGATAATCAT
IGFBP3-681
CCAGCGCTACAAAGTTGACTA
IGFBP3-711
GAGCACAGATACCCAGAACTT
IGFBP3-770
GCCGTAGAGAAATGGAAGACA
IGFIR-532
GCGGTGTCCAATAACTACATT
IGFIR-1959
CGGCAACCTGAGTTACTACAT
IGFIR-3475
GCCGAAGATTTCACAGTCAAA
IGFIR-2427
GCCTTTCACATTGTACCGCAT
doi: 10.1371/journal.pone.0080544.t001 Sunitinib (Sutent®) is a small molecule inhibitor of the
receptor tyrosine kinases of the VEGF family [7], and it[8][9] is
now the front-line standard of care in metastatic RCC. Other
VEGFR inhibitors, such as sorafenib [10], axitinib [11] and
pazopanib [12] were all reported to be active against ccRCC. However, about 20-30% of the patients do not respond to
VEGFR inhibitor therapies and nearly all will become resistant
[9]. These patients are in urgent need of new and effective
therapies, and identifying new drug targets is a prerequisite
step. Name of shRNA constructs
shRNA sequences
HIF1b-1770
GAGAAGTCAGATGGTTTATTT
HIF2a-1631
CGACCTGAAGATTGAAGTGAT
HIF2a-566
CCATGAGGAGATTCGTGAGAA
VEGF-1137
AGGGCAGAATCATCACGAAGT
VEGF-1500
GCGCAAGAAATCCCGGTATAA
VEGF-1587
GACGTGTAAATGTTCCTGCAA
CCND1-326
GCCCTCGGTGTCCTACTTCAA
CCND1-539
CTCTAAGATGAAGGAGACCAT
CCND1-2322
GCCAGGATGATAAGTTCCTTT
ANGPTL4-786
GCAGAGTGGACTATTTGAAAT
ANGPTL4-1311
GAAGCTTAAGAAGGGAATCTT
EGLN3-692
GTGGCTTGCTATCCGGGAAAT
ENO2-1992
CGCCTGGCTAATAAGGCTTTA
ENO2-1735
CGCACTTTCCACTTCTTCCTT
GLUT1-1598
CCAAAGTGATAAGACACCCGA
GLUT1-455
GCGGAATTCAATGCTGATGAT
GLUT1-2310
GCCACACTATTACCATGAGAA
IGFBP3-633
CCTCCATTCAAAGATAATCAT
IGFBP3-681
CCAGCGCTACAAAGTTGACTA
IGFBP3-711
GAGCACAGATACCCAGAACTT
IGFBP3-770
GCCGTAGAGAAATGGAAGACA
IGFIR-532
GCGGTGTCCAATAACTACATT
IGFIR-1959
CGGCAACCTGAGTTACTACAT
IGFIR-3475
GCCGAAGATTTCACAGTCAAA
IGFIR-2427
GCCTTTCACATTGTACCGCAT
doi: 10.1371/journal.pone.0080544.t001 Other HRGs other than VEGF are important for efficient
tumor growth g
The relative importance of constitutively active HIF pathway
in kidney cancer initiation and maintenance has been well
established in xenograft models [13,14]. Down-regulation of
HIF2α expression in VHL-/- kidney cancer cells did not inhibit
cellular growth under standard cell culture condition. However,
it severely impaired these cells’ ability to form tumors in a
xenograft model [15]. The transcriptional activity of the HIF2α
was shown to be critical for its oncogenic activity [16,17],
suggesting that the HIF-responsive genes were largely
responsible for its ability to promote tumor growth. Consistent
with this, Mxi-1, a c-Myc antagonist, was found to possess
oncogenic activity [18]. Similarly, Oct4, a transcriptional factor
essential for maintaining stem cell pluripotency [19], TGF-α, an
agonist for EGFR [20], and Ror2, a receptor tyrosine kinase
[21,22], were all shown to be induced by HIF2α and promoted
tumor growth of kidney cancer cells. However, in addition to
them, HIF regulates many aspects of cell biology such as cell
cycle progression, metabolism and glucose homeostasis, and
cell signaling. The contributions to tumor growth by HRGs
involved in these processes were not fully explored, so in this
study we studied the contributions to tumor growth by seven
HRGs. We found that some HRGs enhanced tumor growth,
some did nothing, while some were tumor-suppressive. protein concentrations of the lysates. Samples with the same
amount of total protein were boiled with sample buffer before
being resolved by SDS-PAGE and analyzed with standard
western blot techniques. The blots were developed with either
Super Signal Pico substrate (Pierce Biotechnology, Rockford,
IL) or Immobilon Western substrate (Millipore, Billerica, MA). Antibody
against
HIF1611079)waspurchased
fromBDtransductionlaboratories (part of BD Biosciences, San
Jose, CA). Antibodies against phopho-tyrosine PY-100 (9411),
actin (4968), IGFIRβ (3018), and anti-Cyclin D1 antibody
(2926) were from Cell Signaling Technology (Boston, MA). Anti-GLUT1
antibody
(NB300-666)
and
anti-HIF2α
(NB100-132)
antibodies
were
purchased
from
Novus
Biologicals (Littleton, CO). Anti-IGFBP3 (AF-675) was from
R&D Systems (Minneapolis, MN). Antibodies against PARP1
(sc-7150), GAPDH (sc-59540) and Vinculin (sc-73614) were
from Santa Cruz Biotechnology (Dallas, TX). protein concentrations of the lysates. Samples with the same
amount of total protein were boiled with sample buffer before
being resolved by SDS-PAGE and analyzed with standard
western blot techniques. The blots were developed with either
Super Signal Pico substrate (Pierce Biotechnology, Rockford,
IL) or Immobilon Western substrate (Millipore, Billerica, MA). Antibody
against
HIF1611079)waspurchased Short Hairpin RNAs (shRNAs) shRNA
constructs
were
obtained
from
Sigma. The
sequences were listed in Table 1. Cell culture 786-O kidney cancer cells with or without pCDNA3 based
wild type HA-VHL were previously described [15]. The VHL
status and the HIF activity were confirmed by anti-HA and anti-
GLUT1 immunoblots. All the cell lines were maintained in
glutamine-containing DMEM medium supplemented with 10%
Fetal Bovine Serum (FBS) and 1% penicillin and streptomycin. For hypoxia mimetic treatment, either 200μM Deferoxamine
(DFO, an iron chelator) or 20μM Cobalt Chloride (CoCl2, which
replaces iron at the active site of the prolyl hydroxylases) was
added to the cell culture media for twelve hours. The cells were
washed and lysed for further analysis. Introduction ubiquitylation and destruction. There are three alpha subunits
of HIF and for the simplicity they are referred to as HIFα. Under
normoxia (normal oxygen tension), prolyl hydroxylase modifies
HIFα on key proline residues (Pro) [3-5], which serve as a
binding signal to the beta domain of pVHL. pVHL-containing
complex then promotes ubiquitylation on HIFα, which leads to
quick proteasomal degradation. Hypoxia (oxygen deprivation)
or other pathological conditions prevents prolyl hydroxylation,
and HIFα accumulates and forms complex with HIF1β. HIF
complex binds to Hypoxic response element (HRE) and
regulates transcription of HIF-responsive genes. Increased HIF
activity as a result of VHL inactivation increases the expression
of many genes and contributes to renal carcinoma growth. Notably, one of the genes whose expression is increased
following VHL inactivation is VEGF, and VEGF and its receptor
VEGFR are confirmed drug targets in ccRCC [6]. The vast majority of renal cell carcinoma (RCC) cases are of
the clear cell type. It is now known that the inactivation of the
VHL tumor suppressor gene plays a causal role in the
pathogenesis of clear cell renal cell carcinomas (ccRCC). In
sporadic ccRCC tumors, about 70% of them harbor biallelic
inactivation
of
VHL
through
mutation,
deletion,
or
hypermethylation of promoter that leads to the loss of its
expression [1]. In hereditary kidney cancer patients, the
inherited germline mutation in one allele of VHL predisposes
them to earlier onset bilateral kidney cancer. The protein
product of VHL tumor suppressor protein, pVHL, is the
substrate recognition unit of an E3 ubiquitin ligase complex that
also contains Cul2, Elongin C and B, and Rbx1[2]. This
complex targets the alpha subunits of the heterodimeric
transcription
factor
HIF
(Hypoxia-Inducible
Factor)
for November 2013 | Volume 8 | Issue 11 | e80544 1 1 PLOS ONE | www.plosone.org Contributions of HIF Target Genes to Kidney Cancer Kidney cancer treatment and drug-resistance Kidney cancer treatment and drug-resistance Kidney cancer treatment and drug-resistance Results Proline 405 on pBabe-Puro-HA-tagged HIF2α-PA-dTA was
mutated to Alanine, the amino acid residues 24-29 RCRRSK
were mutated to ACAASA in the basic helix-loop-helix domain
which would disrupt DNA binding, and the N-terminal and C-
terminal transactivation domains (amino acid residues 450-572
and 820-870) were deleted. Proline 405 on pBabe-Puro-HA-tagged HIF2α-PA-dTA was
mutated to Alanine, the amino acid residues 24-29 RCRRSK
were mutated to ACAASA in the basic helix-loop-helix domain
which would disrupt DNA binding, and the N-terminal and C-
terminal transactivation domains (amino acid residues 450-572
and 820-870) were deleted. Validation of HIF-target genes in 786-O cells In order to examine the relative contributions of individual
HIF-responsive genes (HRG) to the tumor growth, we wished
to validate the HRGs that are tightly regulated by HIF’s
transcriptional activity in 786-O cells before choosing the
targets among them. In VHL-/- cells, the expression of a HRG
should be significantly reduced after HIF2α expression is
suppressed by shRNA. In VHL+/+ cells (where HA-VHL was
stably expressed in the VHL-/- cells), HRG’s expression should
be significantly enhanced by a stable and functional HIF2α
mutant but not by an inactive one. We confirmed that two
independent HIF2α shRNA constructs significantly reduced
HIF2α mRNA in VHL-/- cells, and the VHL+/+ cells with stably
integrated plasmids expressing HIF2a-dPA (active) and the
HIF2a-PA-dTA (inactive) mutants greatly increased the total
HIF2α mRNA levels (Figure 2A). VEGF ELISA Protein concentration of VEGF was measured by an Elisa kit
(Human VEGF Quantikine ELISA kit; R&D Systems) according
to the manufacturer’s instruction. Briefly, 200μl of standard,
control or sample was added to each well and incubated for 2
hours. After three washes, 200μl of VEGF conjugate was
added for a further 2 hours. After washing, 200μl of substrate
solution was added to each well. After twenty minutes
incubation, the reaction was stopped with 50μl stop solution
and the absorbance was read at 450 nm. The experiments
were performed in duplicates. Plasmids Prolines 405 and 531 on pBabe-Puro-HA-tagged HIF2α-dPA
were mutated to Alanines to escape VHL recognition. To
destruct the transcriptional activity of the stabilized HIF2α, The cells were washed before being lysed with EBC buffer
(50 mM Tris (pH 8), 120 mM NaCl, 0.5% NP-40). A protein
assay kit (500-0006) from Bio-rad was used to determine the PLOS ONE | www.plosone.org November 2013 | Volume 8 | Issue 11 | e80544 2 Contributions of HIF Target Genes to Kidney Cancer A functional HIF is essential for tumor growth by VHL-
deficient kidney cancer cells The transcriptional factor Hypoxia Inducible Factor (HIF) is
constituted of two subunits: the labile HIFα and the stable
HIFβ. pVHL targets HIFα for degradation, so HIFα protein
stabilizes and becomes constitutively active in VHL-defective
ccRCC cancer cells [2]. The absence of VHL, which leads to
high level of HIF2α, did not affect the cell growth rates in petri
dishes (Figure S1A in File S1). However, high-level expression
of HIF2α protein has been shown to be necessary and
sufficient for 786-O ccRCC cancer cells to grow into tumors in
orthotopic
and
subcutaneous
xenograft
mouse
models
[13,15-17,23]. To reconfirm the importance of HIF to tumor
growth in a xenograft model, the expression of HIF1β protein in
786-O cells was stably suppressed by an shRNA construct
HIF1b-1770 (Figure 1A). Suppression of HIF1β’s expression in
786-O cells also eliminated the expression of GLUT1, a marker
of HIF’s transcriptional activity (Figure S2 in File S1). As
expected, the inhibition of HIF1β expression significantly
impaired the tumor growth by 786-O cells when compared to
the same cells carrying an shRNA construct expressing a
control sequence that does not target any known gene (SCR)
(Figure 1B and C). Consistent with the previously reported
importance of HIF2α to tumor growth [15], suppression of the
other HIF subunit, HIF2α, also decreased GLUT1 expression
and significantly impaired the tumor growth (Figure 1D, E and
F). Real-time RT-PCR Total RNA was extracted from cells with Trizol reagent
(Invitrogen) following the instructions from the manufacturer. RNA concentration was determined by absorbance at 260nm. A First-strand cDNA Synthesis kit was used to generate First-
strand cDNA (Origene). qRT-PCR was performed using the
7500HT Fast Real-time PCR System (Applied Biosystems) or
with
RT2
Real-Time™
ROX
PCR
Master
Mix
from
SABiosciences. Genes were amplified using the primers
described in Table S1. All quantifications were normalized to β-
actin. In vitro proliferation assays In vitro cell proliferation assays were performed using a Cell
Proliferation Kit II (XTT) from Roche Diagnostics following the
manufacturer's instructions. Nude mouse xenograft assays and statistical analysis Left: Tumor from cells expressing SCR; Right: Tumor from cells expressing HIF1b-1770. D. Total cell lysates
of 786-O cells stably expressing the SCR or the HIF2a-566 were immunoblotted with the indicated antibodies. E. 786-O VHL-/- cells
infected to produce SCR or HIF2a-566 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later,
tumors were excised and weighed. Ten tumors per line were analyzed. Error bars = standard error of the mean. P<=0.001
according to a Mann-Whitney U statistic analysis. F. Representative photographs of tumors analyzed in Figure 1E. Left: Tumor from
cells expressing SCR; Right: Tumor from cells expressing HIF2a-566. doi: 10.1371/journal.pone.0080544.g001 tumor growth by VHL-deficient RCC cells. A. Total cell lysates of human renal carcinoma 786-O
ontrol shRNA (SCR) or the HIF1b-1770 were prepared and immunoblotted with the indicated
ells infected to produce SCR or HIF1b-1770 were injected subcutaneously into the flanks of nude
ks later, tumors were excised and weighed. Thirteen tumors per line were analyzed. Error bars =
<=0.001 according to a Mann-Whitney U statistic analysis. C. Representative photographs of tumors
mor from cells expressing SCR; Right: Tumor from cells expressing HIF1b-1770. D. Total cell lysates
g the SCR or the HIF2a-566 were immunoblotted with the indicated antibodies. E. 786-O VHL-/- cells
F2a-566 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later,
ghed. Ten tumors per line were analyzed. Error bars = standard error of the mean. P<=0.001
U statistic analysis. F. Representative photographs of tumors analyzed in Figure 1E. Left: Tumor from
umor from cells expressing HIF2a-566. Figure 1. HIF is essential to tumor growth by VHL-deficient RCC cells. A. Total cell lysates of human renal carcinoma 786-O
cells stably expressing the control shRNA (SCR) or the HIF1b-1770 were prepared and immunoblotted with the indicated
antibodies. B. 786-O VHL-/- cells infected to produce SCR or HIF1b-1770 were injected subcutaneously into the flanks of nude
mice. Approximately 8-10 weeks later, tumors were excised and weighed. Thirteen tumors per line were analyzed. Error bars =
standard error of the mean. P<=0.001 according to a Mann-Whitney U statistic analysis. C. Representative photographs of tumors
analyzed in Figure 1B. Left: Tumor from cells expressing SCR; Right: Tumor from cells expressing HIF1b-1770. D. Total cell lysates
of 786-O cells stably expressing the SCR or the HIF2a-566 were immunoblotted with the indicated antibodies. E. Nude mouse xenograft assays and statistical analysis 786-O VHL-/- cells
infected to produce SCR or HIF2a-566 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later,
tumors were excised and weighed. Ten tumors per line were analyzed. Error bars = standard error of the mean. P<=0.001
according to a Mann-Whitney U statistic analysis. F. Representative photographs of tumors analyzed in Figure 1E. Left: Tumor from
cells expressing SCR; Right: Tumor from cells expressing HIF2a-566. doi: 10.1371/journal.pone.0080544.g001 xenograft model. VEGF and VEGF receptors are the most
important targets in current clinical treatment of RCC [6]. ANGPTL4 is thought to play critical roles in cancer growth and
progression, angiogenesis, and metastasis[24]. Cyclin D1 is a
very important cell cycle regulator and is reported to be
oncogenic in breast cancer [25], and it was also reported to be
a faithful target of HIF2α that might play an important role in
kidney cancer [26]. ENO2 converts 2-phospho-D-glycerate into
phosphoenolpyruvate and is a key component of the glycolytic
pathway that is critical to ENO1-deficient glioblastoma cancer
cells [27]. EGLN3 is one of the most induced HRGs that could hydroxylate the most critical proline residues on the HIFα
proteins and have HIF-independent targets [28-31]. GLUT1 is a
key glucose transporter that imports glucose into cancer cells
[32]. IGFBP3 is an important regulator of Insulin-like growth
factor pathway [33]. Nude mouse xenograft assays and statistical analysis All animal works were performed with strict accordance with
the recommendations in the Guide for the Care and Use of
Laboratory Animals of the National Institute of Health. The
protocol was approved by a Cleveland Clinic Institutional
Animal Care and Use Committee (ARC 08850). The mice were
anesthetized with isoflurane before subcutaneous injection of
cancer cells, and all efforts were made to minimize pain and
suffering. Subcutaneous nude mice xenograft assays were
performed as previously described [13]. 107 viable cells of a
cell line were injected subcutaneously into one flank of a nude
mouse, and the same number of cells of another cell line was
injected into another flank of the same nude mouse. For each
comparison ten mice were injected in two batches. The mice
were sacrificed 8 to 10 weeks after injection, and tumors were
excised and weighed. Results are presented as mean ±
standard error of the mean. Results were evaluated statistically
using Mann-Whitney U statistic analysis or t-test from
SigmaPlot. Next the total RNA was extracted from these cells and real-
time PCR analysis was performed to analyze the expressions
of potentially interesting HRGs previously identified through
microarray analysis. Many genes were confirmed to be bono
fide HRGs that are tightly regulated by HIF’s transcriptional
activity (Figure 2B and Figure S3 in File S1) while some were
not robustly induced by HIF (data not shown). Among the many
validated
HRGs,
we
decided
to
further
analyze
the
contributions to tumor growth by VEGF, Cyclin D1 (CCND1),
ENOLASE2 (ENO2), EGLN3, ANGPTL4, GLUT1, and IGFBP3,
because they are involved in many well-known aspects of HIF
biology but their roles were not directly investigated in a ccRCC PLOS ONE | www.plosone.org November 2013 | Volume 8 | Issue 11 | e80544 3 Contributions of HIF Target Genes to Kidney Cancer Figure 1. HIF is essential to tumor growth by VHL-deficient RCC cells. A. Total cell lysates of human renal carcinoma 786-O
cells stably expressing the control shRNA (SCR) or the HIF1b-1770 were prepared and immunoblotted with the indicated
antibodies. B. 786-O VHL-/- cells infected to produce SCR or HIF1b-1770 were injected subcutaneously into the flanks of nude
mice. Approximately 8-10 weeks later, tumors were excised and weighed. Thirteen tumors per line were analyzed. Error bars =
standard error of the mean. P<=0.001 according to a Mann-Whitney U statistic analysis. C. Representative photographs of tumors
analyzed in Figure 1B. VEGF and Cyclin D1 both positively contribute to tumor
growth VEGF is a potent pro-angiogenic factor that is critical for
tumor-induced neo-angiogenesis and the current frontline anti-
cancer drugs such as sunitinib, pazopanib and sorafinib all PLOS ONE | www.plosone.org November 2013 | Volume 8 | Issue 11 | e80544 4 Contributions of HIF Target Genes to Kidney Cancer Figure 2. Validation of HIF target genes in VHL-deficient RCC cells. A. The mRNA levels of HIF2α in 786-O VHL+/+ a
VHL-/- cells expressing indicated plasmids or shRNA constructs were measured with real-time PCR. B. The mRNA levels of HIF target genes in 786-O VHL+/+ and VHL-/- cells expressing indicated plasmids or shRNA constru
were measured with real-time PCR. doi: 10.1371/journal.pone.0080544.g002 igure 2
Validation of HIF target genes in VHL-deficient RCC cells
A The mRNA levels of HIF2α in 786 O VHL+/+ Figure 2. Validation of HIF target genes in VHL-deficient RCC cells. A. The mRNA levels of HIF2α in 786-O VHL+/+ and
VHL-/- cells expressing indicated plasmids or shRNA constructs were measured with real-time PCR. B. The mRNA levels of HIF target genes in 786-O VHL+/+ and VHL-/- cells expressing indicated plasmids or shRNA constructs
were measured with real-time PCR. doi: 10.1371/journal.pone.0080544.g002 (Figure S1B in File S1). We then compared the tumor growth of
VHL-/- 786-O cells expressing a control shRNA (SCR) with that
of the same cells expressing VEGF-1137. The cells depleted of
VEGF generated much smaller tumors than the control cells in
the same mice (Figure 3B and C), suggesting that HIF-induced
VEGF expression contributed positively to the tumor growth by
VHL-/- cancer cells. inhibits VEGF receptors and they have clearly shown positive
clinical efficacy [6]. However, although sunitinib is generally
very effective in shrinking tumors, the xenograft tumors formed
by the 786-O VHL-/- cell line tend to be only mildly responsive
to sunitinib treatment in mice [34,35], thus it raises the question
whether VEGF is critical for 786-O VHL-/- cells to grow into
tumor efficiently in mice. To address this, we identified an
shRNA construct VEGF-1137 that very efficiently suppressed
the VEGF expression below the level seen in the VHL+/+ cells
(Figure 3A). Like shRNA constructs against HIF, VEGF-1137
did not significantly change the cell growth in the petri dish Cyclin D1 is a cell cycle regulator that promotes cell
proliferation along with many other important functions that
enhances cancer development [36]. VEGF and Cyclin D1 both positively contribute to tumor
growth It is also another HRG that
has been shown to play an important oncogenic role in breast November 2013 | Volume 8 | Issue 11 | e80544 PLOS ONE | www.plosone.org 5 Contributions of HIF Target Genes to Kidney Cancer re 3. VEGF and Cyclin D1 positively contribute to tumor growth by VHL-deficient RCC cells. A. Lysates from 786-O
stably expressing VHL, the SCR or the VEGF shRNAs were prepared and assayed with VEGF ELISA. B. 786-O VHL-/- cells
ed to produce SCR or VEGF-1137 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later,
rs were excised and weighed. Eight tumors per line were analyzed. Error bars = standard error of the mean. P<=0.001
ding to a Mann-Whitney U statistic analysis. C. Representative photographs of tumors analyzed in Figure 3B. Left: Tumor from
expressing SCR; Right: Tumor from cells expressing VEGF-1137. D. Total cell lysates of 786-O cells stably expressing VHL,
CR or the Cyclin D1 shRNAs were immunoblotted with the indicated antibodies. E. 786-O VHL-/- cells infected to produce SCR
CND1-2322 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later, tumors were excised
weighed. Eight tumors per line were analyzed. Error bars = standard error of the mean. P<=0.001 according to a t-test. F. esentative photographs of tumors analyzed in Figure 3E. Left: Tumor from cells expressing SCR; Right: Tumor from cells
ssing CCND1-2322. 1371/journal.pone.0080544.g003 Figure 3. VEGF and Cyclin D1 positively contribute to tumor growth by VHL-deficient RCC cells. A. Lysates from 786-O
cells stably expressing VHL, the SCR or the VEGF shRNAs were prepared and assayed with VEGF ELISA. B. 786-O VHL-/- cells
infected to produce SCR or VEGF-1137 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later,
tumors were excised and weighed. Eight tumors per line were analyzed. Error bars = standard error of the mean. P<=0.001
according to a Mann-Whitney U statistic analysis. C. Representative photographs of tumors analyzed in Figure 3B. Left: Tumor from
cells expressing SCR; Right: Tumor from cells expressing VEGF-1137. D. Total cell lysates of 786-O cells stably expressing VHL,
the SCR or the Cyclin D1 shRNAs were immunoblotted with the indicated antibodies. E. 786-O VHL-/- cells infected to produce SCR
or CCND1-2322 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later, tumors were excised
and weighed. Eight tumors per line were analyzed. VEGF and Cyclin D1 both positively contribute to tumor
growth Error bars = standard error of the mean. P<=0.001 according to a t-test. F. Representative photographs of tumors analyzed in Figure 3E. Left: Tumor from cells expressing SCR; Right: Tumor from cells
expressing CCND1-2322. Suppression of neither ANGPTL4, EGLN3, nor
ENOLASE 2 significantly change the tumor growth Representative photographs of tumors analyzed in Figure 4B. Figure 4. ENO2, EGLN3, and ANGPTL4 make no significant contribution to tumor growth by VHL-deficient RCC cells. A. The mRNA levels of ANGPTL4 in 786-O cells stably expressing VHL, the SCR or the ANGPTL4 shRNAs were analyzed with real-
time PCR. B. 786-O VHL-/- cells infected to produce SCR or ANGPTL4-786 were injected subcutaneously into the flanks of nude
mice. Approximately 8-10 weeks later, tumors were excised and weighed. Nine tumors per line were analyzed. Error bars =
standard error of the mean. P=0.255 according to a t-test. C. Representative photographs of tumors analyzed in Figure 4B. Left:
Tumor from cells expressing SCR; Right: Tumor from cells expressing ANGPTL4-786. D. The mRNA levels of EGLN3 in 786-O
cells stably expressing VHL, the SCR or the EGLN3 shRNA were analyzed with real-time PCR. E. 786-O VHL-/- cells infected to
produce SCR or EGLN3-692 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later, tumors
were excised and weighed. Seven tumors per line were analyzed. Error bars = standard error of the mean. P=0.097 according to a
Mann-Whitney U statistic analysis. F. Representative photographs of tumors analyzed in Figure 4E. Left: Tumor from cells
expressing SCR; Right: Tumor from cells expressing EGLN3-692. G. The mRNA levels of ENO2 in 786-O cells stably expressing
VHL, the SCR or the ENO2 shRNAs were analyzed with real-time PCR. H. 786-O VHL-/- cells infected to produce SCR or
ENO2-1992 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later, tumors were excised and
weighed. Nine tumors per line were analyzed. Error bars = standard error of the mean. P=0.176 according to a t-test. I. Representative photographs of tumors analyzed in Figure 4H. Left: Tumor from cells expressing SCR; Right: Tumor from cells
expressing ENO2-1992. Figure 4. ENO2, EGLN3, and ANGPTL4 make no significant contribution to tumor growth by VHL-deficient RCC cells. A. The mRNA levels of ANGPTL4 in 786-O cells stably expressing VHL, the SCR or the ANGPTL4 shRNAs were analyzed with real-
time PCR. B. 786-O VHL-/- cells infected to produce SCR or ANGPTL4-786 were injected subcutaneously into the flanks of nude
mice. Approximately 8-10 weeks later, tumors were excised and weighed. Nine tumors per line were analyzed. Error bars =
standard error of the mean. P=0.255 according to a t-test. C. Representative photographs of tumors analyzed in Figure 4B. Suppression of neither ANGPTL4, EGLN3, nor
ENOLASE 2 significantly change the tumor growth cancer [25]. As it is a faithful target for HIF2α in RCC cells [26],
it might also have a significant impact on RCC biology. To
investigate its role in tumor growth of kidney cancer, its
expression in VHL-/- cells were very efficiently knocked down
by CCND1-2322 (Figure 3D). CCND1-2322 did not significantly
change the cell growth in petri dish (Figure S1C in File S1). However, these cells grew into much smaller tumors than the
cells expressing SCR (Figure 3E and F), which strongly
suggests that Cyclin D1, like VEGF, also promoted tumor
growth by the VHL-/- cancer cells in mice. cancer [25]. As it is a faithful target for HIF2α in RCC cells [26],
it might also have a significant impact on RCC biology. To
investigate its role in tumor growth of kidney cancer, its
expression in VHL-/- cells were very efficiently knocked down
by CCND1-2322 (Figure 3D). CCND1-2322 did not significantly
change the cell growth in petri dish (Figure S1C in File S1). However, these cells grew into much smaller tumors than the
cells expressing SCR (Figure 3E and F), which strongly
suggests that Cyclin D1, like VEGF, also promoted tumor
growth by the VHL-/- cancer cells in mice. Recent discoveries revealed that ANGPTL4 was involved in
angiogenesis, altered redox regulation, and metastasis [24], so
we decided to investigate its role in kidney cancer progression. The mRNA level of ANGPTL4 in VHL-/- cells was twice as
much as that in VHL+/+ cells, and the shRNA construct
ANGPTL4-786 canceled that increase in the VHL-/- cells
(Figure 4A). ANGPTL4-786 did not significantly change the cell
growth in petri dish (Figure S1D in File S1). ANGPTL4-786 also
failed to change the tumor growth by VHL-/- cells significantly
(Figure 4B and C). November 2013 | Volume 8 | Issue 11 | e80544 PLOS ONE | www.plosone.org 6 Contributions of HIF Target Genes to Kidney Cancer gure 4. ENO2, EGLN3, and ANGPTL4 make no significant contribution to tumor growth by VHL-deficient RCC cell
e mRNA levels of ANGPTL4 in 786-O cells stably expressing VHL, the SCR or the ANGPTL4 shRNAs were analyzed with
e PCR. B. 786-O VHL-/- cells infected to produce SCR or ANGPTL4-786 were injected subcutaneously into the flanks of
ce. Approximately 8-10 weeks later, tumors were excised and weighed. Nine tumors per line were analyzed. Error b
ndard error of the mean. P=0.255 according to a t-test. C. IGF1R is tumor-promoting in 786-O VHL-/- cells IGFBP3 belongs to a family of proteins that bind to insulin
like growth factors and either inhibit or stimulate the IGF
receptors. In order to investigate whether IGFIR activity is
affected by IGFBP3 depletion in 786-O cells, we probed the
tyrosine phosphorylation signals on IGFIR. We found that
depletion of IGFBP3 did not visibly change the tyrosine
phosphorylation signals associated with IGFIR, suggesting that
there is little change of IGFIR activity (Figure S4 in File S1). We then sought to examine whether IGFIR plays an
oncogenic role in our model. Indeed IGFIR-1959 that effectively
reduced IGFIRβ expression very significantly reduced tumor
growth (Figure 6A, B and C). When we investigated the mutual
interaction between IGFIR and IGFBP3, we made a surprising
discovery that IGFIRβ suppression led to profound up-
regulation of the protein levels of IGFBP3 (Figure 6D). As
IGFIR-532, IGFIR-1959, and IGFIR-3475 reduced IGFIR
protein expression to various extents, and all of them caused
IGFBP3 up-regulation, this was unlikely to be an off-target side
effect of a specific shRNA construct. The measurement of
mRNA levels of IGFIR and IGFBP3 further revealed that IGFIR
loss caused the increase of IGFBP3 mRNA levels, which at
least partially explained the increases of IGFBP3 protein levels
(Figure 6E). It remains a distinct possibility that the strong
tumor suppressive effect of IGFIR loss was mostly due to the
high levels of IGFBP3. If this is the case, since IGFIR is
depleted, IGFBP3 must act in an IGFIR-independent manner. Enolase 2 (ENO2) is an enzyme that hydrolyses 2-phospho-
D-glycerate into phosphoenolpyruvate, a critical step in the
glycolysis pathway that converts glucose into pyruvate and
generates ATPs without using molecular oxygen. It was
reported that in VHL-defective kidney cancer cells the elevated
HIF activity caused the decrease of the amount and repression
of the function of mitochondria and shifted the cellular
metabolism to glycolysis [39], so we wanted to examine the
contribution of ENO2, which is critical to glycolysis, to tumor
growth. The mRNA level of ENO2 was much higher in VHL-/-
cells than that in VHL+/+ cells, and the ENO2-1992 shRNA
construct efficiently reduced its expression in VHL-/- cells
(Figure 4G), but it did not significantly change the cell growth in
petri dish (Figure S1F in File S1). We found that ENO2-1992
failed to change tumor growth significantly (Figure 4H and I). Suppression of HIF-induced genes GLUT1 or IGFBP3
strongly enhance tumor growth The constitutively high level of HIF activity shifts the VHL-
defective kidney cancer cells metabolism from oxidative
phosphorylation toward glycolysis to derive ATP without using
molecular oxygen [39]. However, the efficiency of ATP
generation through glycolysis is much lower than oxidative
phosphorylation, thus the cancer cells demand for glucose is
higher than that of the normal cells. To achieve that, HIF
strongly induces the expression of GLUT1, a glucose
transporter that enables the cancer cells to import glucose
efficiently. To address the importance of GLUT1 to tumor
growth, we successfully eliminated the GLUT1 expression by
stably expressing GLUT1-2310 in these cells (Figure 5A). GLUT1-2310 did not significantly change the cell growth in petri
dish (Figure S1G in File S1). Surprisingly and contrary to the
hypothesis that GLUT1 was essential to tumor growth, its loss
actually increased tumor growth (Figure 5B and C). The
possible reasons for this observation will be discussed later. IGFBP3 is another HRG that is highly induced by HIF and it
is known to have IGF-dependent and IGF-independent
biological functions that are important for cancer development
[33]. To examine its contribution to tumor growth, we identified
several shRNA constructs that suppressed IGFBP3 protein
expression in VHL-/- cells (Figure 5D). IGFBP3-711 was
chosen for further analysis because it was the most effective
construct. IGFBP3-711 did not significantly change the cell The constitutively high level of HIF activity shifts the VHL-
defective kidney cancer cells metabolism from oxidative
phosphorylation toward glycolysis to derive ATP without using
molecular oxygen [39]. However, the efficiency of ATP
generation through glycolysis is much lower than oxidative
phosphorylation, thus the cancer cells demand for glucose is
higher than that of the normal cells. To achieve that, HIF
strongly induces the expression of GLUT1, a glucose
transporter that enables the cancer cells to import glucose
efficiently. To address the importance of GLUT1 to tumor
growth, we successfully eliminated the GLUT1 expression by
stably expressing GLUT1-2310 in these cells (Figure 5A). GLUT1-2310 did not significantly change the cell growth in petri
dish (Figure S1G in File S1). Surprisingly and contrary to the
hypothesis that GLUT1 was essential to tumor growth, its loss
actually increased tumor growth (Figure 5B and C). The
possible reasons for this observation will be discussed later. Suppression of neither ANGPTL4, EGLN3, nor
ENOLASE 2 significantly change the tumor growth Left:
Tumor from cells expressing SCR; Right: Tumor from cells expressing ANGPTL4-786. D. The mRNA levels of EGLN3 in 786-O
cells stably expressing VHL, the SCR or the EGLN3 shRNA were analyzed with real-time PCR. E. 786-O VHL-/- cells infected to
produce SCR or EGLN3-692 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later, tumors
were excised and weighed. Seven tumors per line were analyzed. Error bars = standard error of the mean. P=0.097 according to a
Mann-Whitney U statistic analysis. F. Representative photographs of tumors analyzed in Figure 4E. Left: Tumor from cells
expressing SCR; Right: Tumor from cells expressing EGLN3-692. G. The mRNA levels of ENO2 in 786-O cells stably expressing
VHL, the SCR or the ENO2 shRNAs were analyzed with real-time PCR. H. 786-O VHL-/- cells infected to produce SCR or
ENO2-1992 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later, tumors were excised and
weighed. Nine tumors per line were analyzed. Error bars = standard error of the mean. P=0.176 according to a t-test. I. Representative photographs of tumors analyzed in Figure 4H. Left: Tumor from cells expressing SCR; Right: Tumor from cells
expressing ENO2-1992. November 2013 | Volume 8 | Issue 11 | e80544 7 PLOS ONE | www.plosone.org 7 Contributions of HIF Target Genes to Kidney Cancer Contributions of HIF Target Genes to Kidney Cancer growth in petri dish (Figure S1H in File S1). Interestingly,
suppression of IGFBP3 expression, instead of retarding tumor
growth, very significantly enhanced the tumor growth (Figure
5E and F). Next we examined whether EGLN3, one of the most highly
induced HRG [17,28], plays any role in tumor growth. Although
normally EGLN1 is the major proline hydroxylase for HIFα
proteins, under certain conditions EGLN3 could also modify
HIFα proteins for proteasomal degradation [37,38]. 786-O cells
have no pVHL, so even if HIFα is proline hydroxylated it would
not affect its protein stability. EGLN3 also was also reported to
have non-HIF substrates and possess HIF-independent
biological activities [29-31]. Surprisingly, the shRNA construct
EGLN3-692 that efficient suppressed the protein expression of
EGLN3 did not reduce the tumor growth (the small increase in
tumor weight after EGLN3 knockdown was not statistically
significant) (Figure 4D, E, and F). EGLN3-692 did not
significantly change the cell growth in petri dish either (Figure
S1E in File S1). November 2013 | Volume 8 | Issue 11 | e80544 Discussion Although VEGF and VEGFR have been proved to be critical
targets for RCC treatment, it was surprising that whether down-
regulation of VEGF in ccRCC cells had any impact on tumor
growth was not directly tested in a xenograft system. Besides it
was discovered before that the tumors formed by 786-O cells
were relatively insensitive to VEGF inhibitor sunitinib [34,35],
so we decided to investigate whether the loss of VEGF
expression would reduce tumor growth in 786-O-derived
tumors. The positive result confirmed that VEGF expression
was still important to tumor growth, but it is not as critical as
HIF itself. The stronger effect of the shRNA than that of
sunitinib could simply be due to the relative constant effect of
shRNA while drug might fail to sufficiently inhibits its target
between doses. As CCND1 was a critical oncogene for breast cancer, we
sought to address its importance in this model. As expected, it
also positively contributed to the tumor growth but its efficacy
was moderate compared to HIF. IGFBP3 is another HRG that is highly induced by HIF and it
is known to have IGF-dependent and IGF-independent
biological functions that are important for cancer development
[33]. To examine its contribution to tumor growth, we identified
several shRNA constructs that suppressed IGFBP3 protein
expression in VHL-/- cells (Figure 5D). IGFBP3-711 was
chosen for further analysis because it was the most effective
construct. IGFBP3-711 did not significantly change the cell Since HIF2α overexpression in VHL+/+ 786-O cells was
sufficient to generate big tumors in the xenograft model, we
wondered whether the overexpression of an HRG could do the
same. The overexpression of neither VEGF nor CCND1 in VHL
+/+ 786-O cells was able to cause robust tumor growth (data PLOS ONE | www.plosone.org November 2013 | Volume 8 | Issue 11 | e80544 8 Contributions of HIF Target Genes to Kidney Cancer Figure 5. GLUT1 and IGFBP3 are tumor suppressive HRGs in 786-O cells. A. Lysates from 786-O cells stably expressing VHL,
the SCR or the GLUT1 shRNAs were prepared and immunoblotted with indicated antibodies. B. 786-O VHL-/- cells infected to
produce SCR or GLUT1-2310 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later, tumors
were excised and weighed. Eight tumors per line were analyzed. Error bars = standard error of the mean. P<=0.001 according to a
Mann-Whitney U statistic analysis. C. Discussion Approximately 8-10 weeks later, tumors
were excised and weighed. Eight tumors per line were analyzed. Error bars = standard error of the mean. P<=0.001 according to a
Mann-Whitney U statistic analysis. C. Representative photographs of tumors analyzed in Figure 5B. Left: Tumor from cells
expressing SCR; Right: Tumor from cells expressing GLUT1-2310. D. Total cell lysates of 786-O cells stably expressing VHL, the
SCR or the IGFBP3 shRNAs were immunoblotted with the indicated antibodies. E. 786-O VHL-/- cells infected to produce SCR or
IGFBP3-711 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later, tumors were excised and
weighed. Ten tumors per line were analyzed. Error bars = standard error of the mean. P<=0.001 according to a Mann-Whitney U
statistic analysis. F. Representative photographs of tumors analyzed in Figure 5E. Left: Tumor from cells expressing SCR; Right:
Tumor from cells expressing IGFBP3-711. doi: 10.1371/journal.pone.0080544.g005 [29,30]
and
have
HIF-dependent
and
HIF-independent
biological activities [31], so we wondered whether EGLN3 loss
would have any impact on tumor growth. The VHL-/- 786-O
cells depleted of EGLN3 grew tumors just as well as the control
cells, suggesting that EGLN3 inhibition alone is not a good way
to suppress kidney tumor growth. Similarly, suppression of
ENO2 expression did not slow down the tumor growth. Either
its function is not critical for tumor growth, or the remaining
ENO1,
or
another
activated
compensatory
pathway
circumvents the loss of ENO2 protein. not shown). This was consistent with the observation from
another lab (Othon Iliopoulos, personal communication). Thus
it seems that although both VEGF and Cyclin D1 were able to
promote tumor growth, neither of which was as powerful as
HIF. Although ANGPTL4 was reported to increase or decrease
tumor growth and angiogenesis depending on the cancer types
[24], it did not seem to be a major player in this model as its
efficient suppression did not alter how fast the tumors grew. EGLN3 was found to be among the most highly induced HRGs,
and it is known that it can biochemically hydroxylate key proline
residues on the HIFα proteins [38] and its depletion led to
HIF2α stabilization [37]. Since in 786-O cells the VHL protein is
absent, this would not lead to the destruction of HIFα protein
and did not obviously change the transcriptional activity of HIF. Discussion Representative photographs of tumors analyzed in Figure 5B. Left: Tumor from cells
expressing SCR; Right: Tumor from cells expressing GLUT1-2310. D. Total cell lysates of 786-O cells stably expressing VHL, the
SCR or the IGFBP3 shRNAs were immunoblotted with the indicated antibodies. E. 786-O VHL-/- cells infected to produce SCR or
IGFBP3-711 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later, tumors were excised and
weighed. Ten tumors per line were analyzed. Error bars = standard error of the mean. P<=0.001 according to a Mann-Whitney U
statistic analysis. F. Representative photographs of tumors analyzed in Figure 5E. Left: Tumor from cells expressing SCR; Right:
Tumor from cells expressing IGFBP3-711. doi: 10.1371/journal.pone.0080544.g005 Figure 5. GLUT1 and IGFBP3 are tumor suppressive HRGs in 786-O cells. A. Lysates from 786-O cells stably expressing VHL,
the SCR or the GLUT1 shRNAs were prepared and immunoblotted with indicated antibodies. B. 786-O VHL-/- cells infected to
produce SCR or GLUT1-2310 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later, tumors
were excised and weighed. Eight tumors per line were analyzed. Error bars = standard error of the mean. P<=0.001 according to a
Mann-Whitney U statistic analysis. C. Representative photographs of tumors analyzed in Figure 5B. Left: Tumor from cells
expressing SCR; Right: Tumor from cells expressing GLUT1-2310. D. Total cell lysates of 786-O cells stably expressing VHL, the
SCR or the IGFBP3 shRNAs were immunoblotted with the indicated antibodies. E. 786-O VHL-/- cells infected to produce SCR or
IGFBP3-711 were injected subcutaneously into the flanks of nude mice. Approximately 8-10 weeks later, tumors were excised and
weighed. Ten tumors per line were analyzed. Error bars = standard error of the mean. P<=0.001 according to a Mann-Whitney U
statistic analysis. F. Representative photographs of tumors analyzed in Figure 5E. Left: Tumor from cells expressing SCR; Right:
Tumor from cells expressing IGFBP3-711. doi: 10.1371/journal.pone.0080544.g005 Figure 5. GLUT1 and IGFBP3 are tumor suppressive HRGs in 786-O cells. A. Lysates from 786-O cel P3 are tumor suppressive HRGs in 786-O cells. A. Lysates from 786-O cells stably expressing VHL, Figure 5. GLUT1 and IGFBP3 are tumor suppressive HRGs in 786-O cells. A. Lysates from 786-O cells stably expressing VHL,
the SCR or the GLUT1 shRNAs were prepared and immunoblotted with indicated antibodies. B. 786-O VHL-/- cells infected to
produce SCR or GLUT1-2310 were injected subcutaneously into the flanks of nude mice. November 2013 | Volume 8 | Issue 11 | e80544 Discussion It is also known that EGLN3 has other hydroxylase targets It was totally unexpected that the suppression of GLUT1
enhanced, instead of suppressed, the tumor growth. This
seemed to disagree with a report that a class of compound,
exemplified by STF-31, could directly bind and inhibit GLUT1
function and selectively kill VHL-deficient RCC cancer cells
[40]. It was further revealed that the STF-31 inhibited glucose PLOS ONE | www.plosone.org November 2013 | Volume 8 | Issue 11 | e80544 9 Contributions of HIF Target Genes to Kidney Cancer Figure 6. IGFIR loss suppresses tumor growth by VHL-defective RCC cells. A. Lysates from 786-O VHL-/- cells expressin
SCR or an IGFIR shRNA were immunoblotted with indicated antibodies. B. 786-O VHL-/- cells infected to produce the SCR o
IGFIR-1959 were injected subcutaneously into the flanks of nude mice. Approximately 9 weeks later, tumors were excised an
weighed. Six tumors per line were analyzed. Error bars = standard error of the mean. P=0.002 according to a Mann-Whitney U
statistic analysis. C. Representative photographs of nude mice and tumors analyzed in Figure 6B. Left: Tumor from cells expressin
SCR; Right: Tumor from cells expressing IGFIR-1959. D. Lysates from 786-O VHL-/- cells expressing SCR or IGFIR shRNAs wer
immunoblotted with indicated antibodies. E. The mRNA levels of IGFIR and IGFBP3 in 786-O VHL-/- cells expressing SCR or IGFIR
shRNAs were analyzed with real-time PCR. F. A model depicting how HIF and HRGs regulate tumor growth in ccRCC. doi: 10.1371/journal.pone.0080544.g006 gure 6. IGFIR loss suppresses tumor growth by VHL-defective RCC cells. A. Lysates from 786-O VHL-/- cells expressi
CR or an IGFIR shRNA were immunoblotted with indicated antibodies. B. 786-O VHL-/- cells infected to produce the SCR
GFIR-1959 were injected subcutaneously into the flanks of nude mice. Approximately 9 weeks later, tumors were excised a
eighed. Six tumors per line were analyzed. Error bars = standard error of the mean. P=0.002 according to a Mann-Whitney
atistic analysis. C. Representative photographs of nude mice and tumors analyzed in Figure 6B. Left: Tumor from cells expressi
CR; Right: Tumor from cells expressing IGFIR-1959. D. Lysates from 786-O VHL-/- cells expressing SCR or IGFIR shRNAs we
mmunoblotted with indicated antibodies. E. The mRNA levels of IGFIR and IGFBP3 in 786-O VHL-/- cells expressing SCR or IGF
hRNAs were analyzed with real-time PCR. F. A model depicting how HIF and HRGs regulate tumor growth in ccRCC. Supporting Information Table S1. The list of the primers used for real-time PCR
experiments in this paper. (DOCX) Table S1. The list of the primers used for real-time PCR
experiments in this paper. File S1. File includes Figures S1, S2, S3, and S4. Figure
S1. VHL status or change of HRG expression in 786-O cells
does not alter in vitro growth rates. 786-O VHL+/+, 786-O
VHL-/- cells (A), 786-O VHL-/- cells expressing either control
shRNA
(SCR)
or
VEGF-1137
(B),
CCND1-2322
(C),
ANGPTL4-786
(D),
EGLN3-692
(E),
ENO2-1992
(F),
GLUT1-2310 (G), IGFBP3-711 (H) were used to compare in
vitro proliferation rates. Figure S2. 786-O cells with the HIF1β
depleted have diminished expression of the HRG GLUT1. 786-
O VHL-/- cells expressing either SCR or HIF1b-1770 were
either untreated or treated with hypoxia mimetics DFO or CoCl2
overnight. The cells were washed, lysed and subjected to
western blots with indicated antibodies. Figure S3. The
confirmation of HRGs in 786-O cells. Total RNAs were
extracted from 786-O VHL+/+, 786-O VHL-/- cells, 786-O
VHL-/- cells expressing two shRNA constructs against HIF2α,
and 786-O VHL+/+ cells expressing either a functional HIF2α
mutant or a non-functional HIF2α mutant. First strand cDNA
was generated from these samples then analyzed by real-time
PCR for the indicated genes of interest. Figure S4. IGFBP3
suppression
does
not
lead
to
increase
of
tyrosine
phosphorylation on IGFIR in 786-O cells. Lysates from 786-O
cell stably expressing either SCR or IGFBP3 shRNAs were
used
for
immunoprecipitation
of
IGFIRβ. The
immunoprecipitates
were
immunoblotted
with
indicated
antibodies. Our previous finding that the suppression of JARID1C, an
HRG, actually enhanced tumor growth suggested the existence
of other tumor-inhibitory HRGs [41]. We found that the
suppression of IGFBP3 very significantly increased the tumor
growth, suggesting that it is another potent tumor-inhibitory
HRG. As it binds to IGF which is an activator of IGFIR, a known
oncogene in many types of human cancer cells [42], we
investigated IGFIR activation status after IGFBP3 depletion. No
obvious activation of IGFIR was discovered, suggesting its
tumor-inhibitory activity might be IGFIR independent. We
further investigated whether IGFIR suppression did reduce
tumor growth, and the result confirmed that the expression of
IGFIR was essential for efficient tumor growth. Surprisingly,
loss of IGFIR increased both the mRNA level, and to a greater
extent, the protein level of IGFBP3, further suggesting that the
elevated IGFBP3 might be capable of suppressing tumor
growth in the absence of IGFIR. Supporting Information As IGFBP3 were reported to
interact with TGF-β pathway [43,44], increase the ratio of
apoptotic (Bax and Bad) to the anti-apoptotic proteins (Bcl2
and Bcl-XL) [45], inhibit NF-κB activity [46], its abilities to
inhibits
cell
growth/promotes
apoptosis
in
an
IGFIR-
independent manner were well documented and might be at
play here [33]. So IGFIR could be a very good drug target in
addition to VEGFR in RCC, and any event that significantly
enhances IGFBP3 expression might be desirable when treating
RCC. Acknowledgements The 786-O cell lines were kind gifts from Dr. William Kaelin Jr. We thank Drs. Janet Houghton, Robert Silverman, Stephen
Peiper for their guidance, encouragement and supports. Our results indicate that there might be other HRGs that
mediate the critical oncogenic activity of HIF in RCC, as VEGF
or Cyclin D1 overexpression failed to drive efficient tumor
growth in VHL+/+ cells as the overexpression of HIF2α did. However, it is also possible that the combined effects of many
tumor-promoting HRGs, including VEGF and Cyclin D1, were
responsible for the tumor-inducing power of HIF (Figure 6F). It
is an open question whether the overexpression of a single
HRG is capable of driving efficient tumor growth in VHL+/+ Contributions of HIF Target Genes to Kidney Cancer Contributions of HIF Target Genes to Kidney Cancer uptake, significantly reduced ATP synthesis, led to cell death
and inhibited tumor growth in xenograft models [40]. But a
major difference exists between our studies: we only
suppressed GLUT1 expression, while STF-31 potentially could
inhibit all glucose transporter’s activity. Since GLUT1, GLUT2,
GLUT3, GLUT4 are all expressed at different levels in RCC
and they are all capable of transporting glucose [40], it is highly
possible that in our system the loss of GLUT1 led to
compensatory up-regulation of another glucose transporter
which led to worse outcome, while STF-31 was able to inhibit
the whole family of glucose transporters to inhibit tumor growth. Indeed in RCC4 VHL+/+ cells where the GLUT1 expression
was much lower than that in the RCC4 VHL-/- cells, the GLUT2
level in the VHL+/+ cells was much higher than that in the
VHL-/- cells, presumably due to a compensatory mechanism
[40]. Whatever the explanation, our result indicates that the
inhibition of GLUT1 alone is highly unlikely to be a successful
therapeutic strategy against RCC, while the ability to inhibit all
glucose transporters might be the key to achieve clinical
efficacy. 786-O cells as HIF2α could. To further complicate the matter,
the contributions of HIF-induced microRNAs might also be
important as well, and further research might reveal whether
one or more HRGs or microRNAs, in addition to VEGF and
other important HRGs, might channel the major oncogenic
activity of HIF that can be targeted for efficient therapeutic
intervention in RCC. Discussion i: 10 1371/journal pone 0080544 g006 Figure 6. IGFIR loss suppresses tumor growth by VHL-defective RCC cells. A. Lysates from 786-O VHL-/- cells expressing
SCR or an IGFIR shRNA were immunoblotted with indicated antibodies. B. 786-O VHL-/- cells infected to produce the SCR or
IGFIR-1959 were injected subcutaneously into the flanks of nude mice. Approximately 9 weeks later, tumors were excised and
weighed. Six tumors per line were analyzed. Error bars = standard error of the mean. P=0.002 according to a Mann-Whitney U
statistic analysis. C. Representative photographs of nude mice and tumors analyzed in Figure 6B. Left: Tumor from cells expressing
SCR; Right: Tumor from cells expressing IGFIR-1959. D. Lysates from 786-O VHL-/- cells expressing SCR or IGFIR shRNAs were
immunoblotted with indicated antibodies. E. The mRNA levels of IGFIR and IGFBP3 in 786-O VHL-/- cells expressing SCR or IGFIR
shRNAs were analyzed with real-time PCR. F. A model depicting how HIF and HRGs regulate tumor growth in ccRCC. doi: 10.1371/journal.pone.0080544.g006 November 2013 | Volume 8 | Issue 11 | e80544 10 PLOS ONE | www.plosone.org References Marangoni E, Vincent-Salomon A, Auger N, Degeorges A, Assayag F
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12209156. 23. Zimmer M, Doucette D, Siddiqui N, Iliopoulos O (2004) Inhibition of
hypoxia-inducible factor is sufficient for growth suppression of VHL-/-
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targeted for VHL-mediated destruction by proline hydroxylation:
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science.1059817. PubMed: 11292862. 24. Tan MJ, Teo Z, Sng MK, Zhu P, Tan NS (2012) Emerging roles of
angiopoietin-like 4 in human cancer. Mol Cancer Res 10: 677-688. doi:
10.1158/1541-7786.MCR-11-0519. PubMed: 22661548. 4. Jaakkola P, Mole DR, Tian YM, Wilson MI, Gielbert J et al. (2001)
Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex
by O2-regulated prolyl hydroxylation. Science 292: 468-472. doi:
10.1126/science.1059796. PubMed: 11292861. 25. Yu Q, Geng Y, Sicinski P (2001) Specific protection against breast
cancers by cyclin D1 ablation. Nature 411: 1017-1021. doi:
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regulated by stimulus-sensitive proline hydroxylation. Proc Natl Acad
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11504942. 26. Raval RR, Lau KW, Tran MG, Sowter HM, Mandriota SJ et al. (2005)
Contrasting properties of hypoxia-inducible factor 1 (HIF-1) and HIF-2
in von Hippel-Lindau-associated renal cell carcinoma. Mol Cell Biol 25:
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endothelial growth factor in metastatic renal cell carcinoma: biology,
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10.1111/j.1464-410X.2005.05616.x. PubMed: 16042715. 27. Muller FL, Colla S, Aquilanti E, Manzo VE, Genovese G et al. (2012)
Passenger deletions generate therapeutic vulnerabilities in cancer. Nature 488: 337-342. doi:10.1038/nature11331. PubMed: 22895339. 7. References Maranchie JK, Vasselli JR, Riss J, Bonifacino JS, Linehan WM et al. (2002) The contribution of VHL substrate binding and HIF1-alpha to the
phenotype of VHL loss in renal cell carcinoma. Cancer Cell 1: 247-255. doi:10.1016/S1535-6108(02)00044-2. PubMed: 12086861. 35. Bhatt RS, Wang X, Zhang L, Collins MP, Signoretti S et al. (2010)
Renal cancer resistance to antiangiogenic therapy is delayed by
restoration of angiostatic signaling. Mol Cancer Ther 9: 2793-2802. doi:
10.1158/1535-7163.MCT-10-0477. PubMed: 20699227. (
)
15. Kondo K, Kim WY, Lechpammer M, Kaelin WG Jr. (2003) Inhibition of
HIF2alpha is sufficient to suppress pVHL-defective tumor growth. PLOS Biol 1: E83. PubMed: 14691554. 36. Pestell RG (2013) New roles of cyclin d1. Am J Pathol 183: 3-9. doi:
10.1016/S0002-9440(13)00360-X. PubMed: 23790801. 16. Yan Q, Bartz S, Mao M, Li L, Kaelin WG Jr. (2007) The hypoxia-
inducible factor 2alpha N-terminal and C-terminal transactivation
domains cooperate to promote renal tumorigenesis in vivo. Mol Cell
Biol 27: 2092-2102. doi:10.1128/MCB.01514-06. PubMed: 17220275. 37. Taniguchi CM, Finger EC, Krieg AJ, Wu C, Diep AN et al. (2013)
Cross-talk between hypoxia and insulin signaling through Phd3
regulates hepatic glucose and lipid metabolism and ameliorates
diabetes. Nat Med, 19: 1325–30. PubMed: 24037093. 17. Li L, Zhang L, Zhang X, Yan Q, Minamishima YA et al. (2007) Hypoxia-
inducible factor linked to differential kidney cancer risk seen with type
2A and type 2B VHL mutations. Mol Cell Biol 27: 5381-5392. doi:
10.1128/MCB.00282-07. PubMed: 17526729. 38. Epstein AC, Gleadle JM, McNeill LA, Hewitson KS, O'Rourke J et al. (2001) C. elegans EGL-9 and mammalian homologs define a family of
dioxygenases that regulate HIF by prolyl hydroxylation. Cell 107: 43-54. doi:10.1016/S0092-8674(01)00507-4. PubMed: 11595184. 18. Tsao CC, Teh BT, Jonasch E, Shreiber-Agus N, Efstathiou E et al. (2008) Inhibition of Mxi1 suppresses HIF-2alpha-dependent renal
cancer tumorigenesis. Cancer Biol Ther 7: 1619-1627. doi:10.4161/cbt. 7.10.6583. PubMed: 19018165. 39. Zhang H, Gao P, Fukuda R, Kumar G, Krishnamachary B et al. (2007)
HIF-1 inhibits mitochondrial biogenesis and cellular respiration in VHL-
deficient renal cell carcinoma by repression of C-MYC activity. Cancer
Cell 11: 407-420. doi:10.1016/j.ccr.2007.04.001. PubMed: 17482131. 19. Covello KL, Kehler J, Yu H, Gordan JD, Arsham AM et al. (2006)
HIF-2alpha regulates Oct-4: effects of hypoxia on stem cell function,
embryonic development, and tumor growth. Genes Dev 20: 557-570. doi:10.1101/gad.1399906. PubMed: 16510872. 40. Chan DA, Sutphin PD, Nguyen P, Turcotte S, Lai EW et al. Author Contributions Conceived and designed the experiments: HY. Performed the
experiments: TZ XN LL EAC. Analyzed the data: HY TZ XN. Wrote the manuscript: HY. November 2013 | Volume 8 | Issue 11 | e80544 PLOS ONE | www.plosone.org 11 Contributions of HIF Target Genes to Kidney Cancer 45. Butt AJ, Firth SM, King MA, Baxter RC (2000) Insulin-like growth factor-
binding protein-3 modulates expression of Bax and Bcl-2 and
potentiates p53-independent radiation-induced apoptosis in human
breast cancer cells. J Biol Chem 275: 39174-39181. doi:10.1074/
jbc.M908888199. PubMed: 10998426. human breast cancer cells. J Biol Chem 268: 26045-26048. PubMed:
7504671. Contributions of HIF Target Genes to Kidney Cancer 46. H-Zadeh AM, Collard TJ, Malik K, Hicks DJ, Paraskeva C et al. (2006)
Induction of apoptosis by the 16-kDa amino-terminal fragment of the
insulin-like growth factor binding protein 3 in human colonic carcinoma
cells. Int J Oncol 29: 1279-1286. PubMed: 17016662. human breast cancer cells. J Biol Chem 268: 26045-26048. PubMed:
7504671. 7504671.
45. Butt AJ, Firth SM, King MA, Baxter RC (2000) Insulin-like growth factor-
binding protein-3 modulates expression of Bax and Bcl-2 and
potentiates p53-independent radiation-induced apoptosis in human
breast cancer cells. J Biol Chem 275: 39174-39181. doi:10.1074/
jbc.M908888199. PubMed: 10998426.
46. H-Zadeh AM, Collard TJ, Malik K, Hicks DJ, Paraskeva C et al. (2006)
Induction of apoptosis by the 16-kDa amino-terminal fragment of the
insulin-like growth factor binding protein 3 in human colonic carcinoma
cells. Int J Oncol 29: 1279-1286. PubMed: 17016662. References (2011)
Targeting GLUT1 and the Warburg effect in renal cell carcinoma by
chemical synthetic lethality. Sci Transl Med 3: 94ra70. PubMed:
21813754. 20. Gunaratnam L, Morley M, Franovic A, de Paulsen N, Mekhail K et al. (2003) Hypoxia inducible factor activates the transforming growth
factor-alpha/epidermal growth factor receptor growth stimulatory
pathway in VHL(-/-) renal cell carcinoma cells. J Biol Chem 278:
44966-44974. doi:10.1074/jbc.M305502200. PubMed: 12944410. 41. Niu X, Zhang T, Liao L, Zhou L, Lindner DJ et al. (2012) The von
Hippel-Lindau tumor suppressor protein regulates gene expression and
tumor growth through histone demethylase JARID1C. Oncogene 31:
776-786. doi:10.1038/onc.2011.266. PubMed: 21725364. 776-786. doi:10.1038/onc.2011.266. PubMed: 21725364. 42. Boone DN, Lee AV (2012) Targeting the insulin-like growth factor
receptor: developing biomarkers from gene expression profiling. Crit 21. Wright TM, Brannon AR, Gordan JD, Mikels AJ, Mitchell C et al. (2009)
Ror2, a developmentally regulated kinase, promotes tumor growth PLOS ONE | www.plosone.org 12 November 2013 | Volume 8 | Issue 11 | e80544 November 2013 | Volume 8 | Issue 11 | e80544 Rev
Oncog
17:
161-173. doi:10.1615/CritRevOncog.v17.i2.30. PubMed: 22471706. 45. Butt AJ, Firth SM, King MA, Baxter RC (2000) Insulin-like growth factor-
binding protein-3 modulates expression of Bax and Bcl-2 and
potentiates p53-independent radiation-induced apoptosis in human
breast cancer cells. J Biol Chem 275: 39174-39181. doi:10.1074/
jbc.M908888199. PubMed: 10998426. 43. Oh Y, Müller HL, Lamson G, Rosenfeld RG (1993) Insulin-like growth
factor (IGF)-independent action of IGF-binding protein-3 in Hs578T
human breast cancer cells. Cell surface binding and growth inhibition. J
Biol Chem 268: 14964-14971. PubMed: 7686909. 44. Oh Y, Müller HL, Pham H, Rosenfeld RG (1993) Demonstration of
receptors for insulin-like growth factor binding protein-3 on Hs578T PLOS ONE | www.plosone.org PLOS ONE | www.plosone.org 13 November 2013 | Volume 8 | Issue 11 | e80544
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Qeios · Definition, August 24, 2022 Open Peer Review on Qeios Behaviour Change Intervention Ontology (BCIO) Behaviour Change Intervention Ontology (BCIO) Qeios ID: YYI31K · https://doi.org/10.32388/YYI31K Behaviour Change Intervention Ontology (BCIO) Behaviour Change Intervention Ontology (BCIO) Behaviour Change Intervention Ontology (BCIO) Definition: A behaviour that results from a learnt stimulus-behaviour co-occurrence. This definition was imported from the Behaviour Change Intervention Ontology (see https://bciovocab.org/). Comments
and suggestions for improvements are welcome using the Qeios review system. Definitions imported from the Behaviour Change Intervention Ontology (BCIO) are what are known as ‘ontological
definitions’. See this article in Qeios for an explanation https://www.qeios.com/read/YGIF9B. Ontological definitions can sometimes be hard to read. In those cases we also include an informal definition. Definitions also often require elaboration to make it clear how they should be used and what they include. In those cases
we include a comment. Definitions also often require elaboration to make it clear how they should be used and what they include. In those cases
we include a comment. Also, definitions sometimes require an explanation as to how they came about to help users understand how they relate to
alternative definitions. In those cases we include a curator note. Also, definitions sometimes require an explanation as to how they came about to help users understand how they relate to
alternative definitions. In those cases we include a curator note. Qeios ID: YYI31K · https://doi.org/10.32388/YYI31K 1/1
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Correction: Seeing the �Big� Picture: Big Data Methods for Exploring Relationships Between Usage, Language, and Outcome in Internet Intervention Data (Preprint)
|
Jordan Carpenter
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English
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Spoken
| 1,241
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Correction: Seeing the “Big” Picture: Big Data Methods for
Exploring Relationships Between Usage, Language, and Outcome
in Internet Intervention Data Correction: Seeing the “Big” Picture: Big Data Methods for
Exploring Relationships Between Usage, Language, and Outcome
in Internet Intervention Data Corresponding Author:
Acacia C Parks, PhD
Department of Psychology
Hiram College
Bates Hall 215
11715 Garfield Road
Hiram, OH, 44234
United States
Phone: 1 330 569 5229
Fax: 1 330 569 5398
Email: parksac@hiram edu (J Med Internet Res 2017;19(12):e347) doi: 10.2196/jmir.8099 The authors of, “Seeing the 'Big' Picture: Big Data Methods for
Exploring Relationships Between Usage, Language, and
Outcome in Internet Intervention Data” (J Med Internet Res
2016;18(8):e241) would like to make changes to the following
areas. they used Happify daily than during a week when they
didn’t use it at all.” 2. Related Article: Correction of: http://www.jmir.org/2016/8/e241/ J Med Internet Res 2017 | vol. 19 | iss. 12 | e347 | p. 1
(page number not for citation purposes) Corrigenda and Addenda
Correction: Seeing the “Big” Picture: Big Data Methods for
Exploring Relationships Between Usage, Language, and Outcome
in Internet Intervention Data
Jordan Carpenter1, PhD; Patrick Crutchley1,2, BSE; Ran D Zilca3, MSc; H Andrew Schwartz2,4, PhD; Laura K Smith1,
BA; Angela M Cobb5, BA; Acacia C Parks3,5, PhD
1Positive Psychology Center, University of Pennsylvania, Philadelphia, PA, United States
2Penn Medicine Social Media and Health Innovation Lab, Penn Medicine Center for Health Care Innovation, University of Pennsylvania, Philadelphia,
PA, United States
3Happify, New York, NY, United States
Carpenter et al
JOURNAL OF MEDICAL INTERNET RESEARCH JOURNAL OF MEDICAL INTERNET RESEARCH Carpenter et al Corrigenda and Addenda Jordan Carpenter1, PhD; Patrick Crutchley1,2, BSE; Ran D Zilca3, MSc; H Andrew Schwartz2,4, PhD; Laura K Smith1,
BA; Angela M Cobb5, BA; Acacia C Parks3,5, PhD
1Positive Psychology Center, University of Pennsylvania, Philadelphia, PA, United States
2Penn Medicine Social Media and Health Innovation Lab, Penn Medicine Center for Health Care Innovation, University of Pennsylvania, Philadelphia,
PA, United States
3Happify, New York, NY, United States
4Computer Science, Stony Brook University, Stony Brook, NY, United States
5Department of Psychology, Hiram College, Hiram, OH, United States In the Study 1 Results section a. The last sentence in the second paragraph in the Study
1 Results section “This suggests an average overall
improvement of 11.04 points, or about 11%, over the
course of 8 weeks.” This should be replaced with:
“This suggests an average overall improvement of
11.04 points, or about 27%, over the course of 8
weeks.” 1. In the results section of the abstract 1. In the results section of the abstract a. The first sentence of the results subheading in the
abstract “On a measure of positive emotion, the average
user improved 1.38 points per week (SE 0.01,
t122,455=113.60, P<.001, 95% CI 1.36–1.41), about
a 11% increase over 8 weeks.” This should be
replaced with: “On a measure of positive emotion, the
average user improved 1.38 points per week (SE 0.01,
t122,455=113.60, P<.001, 95% CI 1.36–1.41), about
a 27% increase over 8 weeks.” b. The second sentence in the fourth paragraph in the
Study 1 Results section “This estimate predicted that
a given user would report positive emotion that would
be 1.26 points (or 1.26%) higher after a 2-week period
when they used Happify daily than after a week when
they didn’t use it at all.” This should be replaced with:
“This estimate predicted that a given user would report
positive emotion that would be 1.26 points higher after
a 2-week period when they used Happify daily than
after a week when they didn’t use it at all.” b. The third sentence in the results subheading of the
abstract “This estimate predicted that a given user
would report positive emotion 1.26 points (or 1.26%)
higher after a 2-week period when they used Happify
daily than during a week when they didn’t use it at all.”
This should be replaced with: “This estimate
predicted that a given user would report positive
emotion 1.26 points higher after a 2-week period when Jordan Carpenter1, PhD; Patrick Crutchley1,2, BSE; Ran D Zilca3, MSc; H Andrew Schwartz2,4, PhD; Laura K Smith1,
BA; Angela M Cobb5, BA; Acacia C Parks3,5, PhD J Med Internet Res 2017 | vol. 19 | iss. 12 | e347 | p. 2
(page number not for citation purposes) 3.
In the general Discussion section a. The last sentence in the second paragraph in the Study
1 Results section “This suggests an average overall a. The last sentence in the second paragraph in the Study
1 Results section “This suggests an average overall J Med Internet Res 2017 | vol. 19 | iss. 12 | e347 | p. 1
(page number not for citation purposes) J Med Internet Res 2017 | vol. 19 | iss. 12 | e347 | p. 1
(page number not for citation purposes) http://www.jmir.org/2017/12/e347/ XSL•FO
RenderX JOURNAL OF MEDICAL INTERNET RESEARCH Carpenter et al to the interpretation of the results, as well as accurately represent
the magnitude of the results. improvement of 11.04 points, or about 11%, over the
course of 8 weeks.” This should be replaced with:
“This suggests an average overall improvement of
11.04 points, or about 27%, over the course of 8
weeks.” The corrected article will appear in the online version of the
paper on the JMIR website on December 19, 2017, together
with the publication of this correction notice. Because this was
made after submission to PubMed Central, the corrected article
also has been re-submitted to PubMed Central. While the errors do not impact the general findings of the
original publication, remediation of these errors will add clarity Edited by G Eysenbach; this is a non–peer-reviewed article. Submitted 25.05.17; accepted 20.06.17; published 19 Please cite as:
Carpenter J, Crutchley P, Zilca RD, Schwartz HA, Smith LK, Cobb AM, Parks AC
Correction: Seeing the “Big” Picture: Big Data Methods for Exploring Relationships Between Usage, Language, and Outcome in
Internet Intervention Data
J Med Internet Res 2017;19(12):e347
URL: http://www.jmir.org/2017/12/e347/
doi: 10.2196/jmir.8099
PMID: 29258059 Please cite as:
Carpenter J, Crutchley P, Zilca RD, Schwartz HA, Smith LK, Cobb AM, Parks AC
Correction: Seeing the “Big” Picture: Big Data Methods for Exploring Relationships Between Usage, Language, and Outcome in
Internet Intervention Data
J Med Internet Res 2017;19(12):e347
URL: http://www.jmir.org/2017/12/e347/
doi: 10.2196/jmir.8099
PMID: 29258059 ©Jordan Carpenter, Patrick Crutchley, Ran D Zilca, H Andrew Schwartz, Laura K Smith, Angela M Cobb, Acacia C Parks. Originally published in the Journal of Medical Internet Research (http://www.jmir.org), 19.12.2017. This is an open-access article
distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which
permits unrestricted use, distribution, and reproduction in any medium, provided the original work, first published in the Journal
of Medical Internet Research, is properly cited. 3.
In the general Discussion section The complete bibliographic information, a link to the original publication on
http://www.jmir.org/, as well as this copyright and license information must be included. ©Jordan Carpenter, Patrick Crutchley, Ran D Zilca, H Andrew Schwartz, Laura K Smith, Angela M Cobb, Acacia C Parks. Originally published in the Journal of Medical Internet Research (http://www.jmir.org), 19.12.2017. This is an open-access article
distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which
permits unrestricted use, distribution, and reproduction in any medium, provided the original work, first published in the Journal
of Medical Internet Research, is properly cited. The complete bibliographic information, a link to the original publication on
http://www.jmir.org/, as well as this copyright and license information must be included. http://www.jmir.org/2017/12/e347/ XSL•FO
RenderX
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The wax glands of the cockroach (blatta germanica)
|
E. H. Dusham
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English
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Spoken
| 5,326
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Translation by Dr. Jose Nonidez,
Columbia University. Department of Entomology, Cornell University Department of Entomology, Cornell University Las glhdulas productoras de cera en la cucaracha (Blatta
germanica) . La hipodermis de 10s insectos ha sido descrita como formada
por una sola capa de c6lulas. Una excepcidn se presenta en la
pared dorsal del abdomen del macho de la cucaracha (Periplaneta
orientalis), cuya hipodermis estd formada por dos o tres capas
de c6lulas. El estudio de las paredes del abdomen ha demostrado
que en esta especie dicha estructura existe, no solo en las paredes
dorsal y ventral del macho, sino tambi6n en las de la hembra. El autor ha encontrado tambi6n una estructura semejante en
Blatta germanica. La capa superior estA formada por cklulas
hipod6rmicas semejantes a las que existen en otras regiones del
cuerpo, en las que existen c6lulas hipodkrmicas tipicas. Las
capas m&s profundas estan formadas por c6lulas mucho mayores,
con grandes nficleos vesiculares y citoplasma granuloso. Su
aspecto irregular indica su naturaleza glandular, comprobada
ademas por el hallazgo de canales pequefios que se extienden a
trav6s de la cuticula que recubre a dichas glandulas, la cual
presenta pequefios poros en vista superficial, y por medio de 10s
cuales la secrecidn sale a la superficie. El analisis de dicha
secrecidn, recogida en la cuticula, ha demostrado que esta
formada de cera. En Blatta germanica estas glandulas se dis-
tinguen por primera vez despues de la primera muda del tegu-
mento, en las ninfas de dos dias, en las cuales algunas de las
cblulas hipod6rmicas que ocupan ciertas Areas, aumentan de
tamaiio y empujando a las c6lulas hipod6rmicas adyacentes dan
a la hipodermis el aspecto de una capa poliestratificada. La
hipodermis de la cucaracha, por esta causa, no difiere de la de
10s otros insectos. AUTHOR'S ABSTRACT OF THIS PAPER ISSUED
BY THE RJBLIOQRAPHIC SERVICE, NOYEMBER 18 E. H. DUSHAM Department of Entomology, Cornell University INTRODUCTION The ability to secrete wax in one form or another is wide-
spread among insects, being reported for the Libellulidae of the
Odonata ; Notonectidae, Fulgoridae, Cercopidae, Psyllidae, Aphi-
didae, Aleyrodidae, and Coccidae of the Hemiptera; Tortricidae
and Papilionidae of the Lepidoptera ; Coccinellidae and Cur-
culionidae of the Coleoptera; and the Tenthredinidae and Apidae
of the Hjmenoptera. That this function should also be found in cockroaches is not
surprising, for they are waxy in appearance, feel waxy when
handled, and when thrown into water or other fluids float readily
as if buoyed up by some oily covering, even after the air has
been forced out of the tracheae by pressure: In fact, one of the
difficulties encountered in fixing material is to submerge the
specimens in fixing fluids, especially when these are used cold. On the other hand, no such difficulty is experienced when hot
fluids are used, the hot liquid evidently melting the wax, thus
allowing the insects to sink. Moreover, it is a well-known fact that the species in question
is usually found in warm, damp places, kspecially in close prox-
imity to sweating water pipes, its association with water pipes
leading from the Croton Aqueduct in the vicinity of New York
City earning for it the popular name of 'Croton bug.' Further-
more, references to the literature of this species show that it has
been carried for long distances in water mains without drown- 563 564 E. H. DUSHAM ing. That they have been able to live in moist places and also
withstand submerging would lead one to suppose that their
bodies were protected by some oily or waxy secretion. HISTORICAL The study of the wax glands of the cockroach is bound up in
a controversy as to the number of layers in the hypodermis of
these insects. In insects in general the body wall is relatively
simple, consisting of an outer chitinous covering or cuticula, the
hypodermis consisting of a single layer of cells, and the basement
membrane, and as such, Miall and Denny described it in their
work on the cockroach, in which they stated that the hypodermis
consisted of a single layer of flattened cells, resting on a basement
mebrane, each cell corresponding to a polygonal area of the
overlying cuticula. y g
In 1888, however, E. A. Minchin, while studying certain
glands on the dorsal side of the cockroach, found that the hypo-
dermis, at least on the dorsal side of the older stages of the male,
consisted of two layers everywhere except in the intersegmental
areas where only a single layer was present. In certain places
he found that the cells of these layers had the appearance of
giant cells with large nuclei, granular contents, and elongate
processes. These he interpreted as ganglion cells. He found
them scattered over each tergum, especially at the anterior por-
tion of each tergite-that
part which is overlapped by the pre-
ceding tergite-although
they were also scattered throughout
the region posterior to this part. Between these so-called gang-
lion cells he found the ordinary cells of the hypodermis. He
therefore concluded that in the cockroach, at least, the hypo-
dermis consisted of an upper layer of cells corresponding to the
polygonal areas of the cuticula, and a lower very irregular layer,
occasionally forming two layers, whose cells were modified to
form nerve cells, and which were probably connected with setae
where the terga were exposed. g
p
In 1889, Mingazzini made a detailed study of this apparently
double-layered hypodermis in order to verify Minchin’s work, WAX GLANDS OF THE COCKROACH 565 examining the dorsal wall of the abdomen of males in different
stages of development. He found that the hypodermis was not
always made up of two layers of cells as Minchin described, but
sometimes showed a single layer of cells, other times two or more
layers. In the intersegmental areas, he found a single-layer of
cells as Minchin described. HISTORICAL Where the muscle bundles were
inserted he also found a single layer of cells, modified in the usual
way to form the characteristic muscle attachments. In other
places the hypodermis consisted of a single layer of cells, usualiy
of small size, or of two layers, the upper layer consisting of small
cells, the lower one of much larger cells, or of several layers of
cells arranged without order. Except for an occasional large
cell at the surface among the smaller cells where a singlelayer
of cells was present, the majority of the larger cells were below
the smaller ones. He concluded that these cells were not to be considered as
nerve cells, but as cells of an epithelial nature, derived from the
upper layer of the hypodermis, but which had undergone en-
largement and were no longer of use in the secretion of the cu-
ticula. He was uncertain whether these large cells were hair-
forming cells or gland cells, but because of the structure of the
nucleus and protoplasm he favored the glandular view, the in-
sects probably secreting an oily substance. He, therefore, con-
cluded that the hypodermis of the cockroach did not differ from
that of other insects by the nature of the cells which composed
it, but only because, as the insects increase in size, many hypo-
dermal cells become specialized for a particular function, in-
creased in size, took on a branching form, and were carried below
the regular hypodermis, thus forming an apparently double-
layered hypodermis. In 1909, Berlese, in commenting on this peculiar structure in
the cockroach, stated that if the smaller cells were not merely
infiltrated amoebocytes, they were to be regarded as a prolifera-
tion of the hypodermis, and that possibly the larger cells had
assumed another function. At least, he was sure that the con-
dition was unique among insects and was still an open question. 566 E. H. DUSHAM LOCATION AND STRUCTURE OF THE GLANDS At the suggestion of Dr. W A. Riley, under whose direction
this work was carried out, the writer undertook to settle this
question. While the preceding investigations were made on
Periplaneta orientalis, yet a study of Blatta germanica showed
that similar conditions were present, and because of the abun-
dance of the latter, this species was used almost entirely in the
following work. g
In order to show the general distribution of the glands, the
abdomen was cut from the anterior part of the body, and a slit
through the chitin was made all around its margin, thus sepa-
rating the dorsal from the ventral wall. From these respective
parts the fat and muscle were removed as carefully as possible
so as not to injure the delicate hypodermis. It was found more
advisable to fix the entire abdomen before making the incision
around the edge, because the greater rigidity after fixation facili-
tated cutting, and prevented the parts from curling, as when
fresh material was cut. Both dorsal and ventral abdominal walls
were then stained with Grenacher’s borax carmine and Delafield’s
haematoxylin, and mounted in balsam. Better results were ob-
tained with the latter stain. The gland cells are so small that
they can hardly be distinguished from the normal hypodermis
except for the larger size of their nuclei, and these were rendered
more conspicuous by using Delafield’s haematoxylin and destain-
ing with acid alcohol until the nuclei stood out prominently,
while the surrounding cytoplasm of the cells was but faintly
tinged. The nuclei are even better accentuated by dipping the
material in alkaline alcohol after destaining in acid alcohol, as
they take on a deep blue coloration after such treatment. y
p
As the glands are most prominent at the anterior portion of
each segment, and as this portion is covered by the posterior
part of the preceding segment, some difficulty was encountered
in making out their distribution. However, by injecting the ab-
domen with killing fluid by means of a hypodermic needle so
that the intersegmental membranes were stretched taut on a plane
with the rest of the tergite or sternite, all the glandular areas were 567 WAX GLANDS OF THE COCKROACH exposed. Bouin’s fluid proved the most practical for this pur-
pose. LOCATION AND STRUCTURE OF THE GLANDS When the parts were fully extended, they were plunged
into hot killing fluid and the pressure maintained until the hot-
killing fluid without and the killing fluid within had so fixed the
material that the parts would remain extended when the pressure
from the hypodermic needle was released. yp
For histological study the material was fixed in Fleming’s
(strong fluid) , Gilson’s, Zenker’s, Bouin’s, and Dietrich’s fluids,
Best cytological results were obtained with Fleming’s, although
Dietrich’s and Bouin’s fluids gave almost as good results. With
both of the latter‘ more rapid penetration certainly was obtained. Gilson’s and Zenker’s fluids gave almost similar results. g
At first, material was imbedded entirely in paraffin, but when
sections were cut, it was found that the chitin was very brittle
and broke very readily, often catching on the edge of the knife
and tearing gaps through the entire section. Because of this,
sections were so mutilated that they were often useless, espe-
cially as the parts desired for study were immediately under the
chitin. To obviate this difficulty several methods were tried
either to soften the chitin or else to hold the parts in place so
that they would maintain their normal position as nearly as pos-
sible. Metalinkoff’s method, by which the object was first h-
bedded in paraffin in the usual way, and then all the paraffin
scraped from the surface of the chitin, the object immersed in
Eau de Javelle for twenty-four hours, and re-imbedded, did not
give good results. But as this method was tried in only a few
cases perhaps repeated trials would have proved more successful. Bedau’s ‘Seifenspiritus’ method gave no results, although fol-
lowed as exactly as possible. The combined paraffin and celloidin method was also used. Three different modifications of this method were employed-
Gilson’s rapid process, Apathy’s method, and Hoffman’s method. All three gave good results, and longitudinal sections through
the entire abdomen, 5 micra thick, were easily obtained. y
Cross and longitudinal sections of the entire abdomen were
cut from 4 to 10 micra in thickness. .These were fixed to slides
by means of the Mayer albumen-and-water method. Instead, 568 E. H. LOCATION AND STRUCTURE OF THE GLANDS DUSHAM however, of first rubbing a drop of albumen on the slide and then
adding water, three drops of the albumen were added to a watch-
glass of distilled water, thoroughly mixed with it, and a layer of
this was applied to the slide with a pipette. Sections were
then placed on this and flattened by means of heat. By this
method the least amount of albumen necessary remains on the
slide, but always enough to hold the sections firmly in place. y
g
y
p
Sections were stained with Heidenhain’s iron haematoxylin
and Delafield’s haematoxylin with eosin as a counterstain. Bet-
ter cytological results were ,obtained with the former, especially
after fixation with Fleming’s fluid, although good results were
also obtained after fixation with Bouin’s fluid. However, for
general histological work, Delafield’s haematoxylin and eosin
gave very good preparations. g
y g
p p
Prepared mounts of the entire dorsal and ventral walls, sup-
plemented by cross and longitudinal sections of the entire abdo-
men, showed that these modified hypodermal cells were present
in each segment, not only on the dorsal side of the male, but on
the ventral side also, and in the female as well as the male (figs. 1, 2, 3, and 4). Except in the intersegmental membranes, they
are scattered over each tergite and sternite, being extremely
abundant at the fore part of each segment laterad of the median
line, especially that part of the segment which is overlapped by
the preceding segment. Posterior to this region, i.e., where the
tergite or sternite is not covered by the preceding segment, these
modified cells are more scattered, occurring here and there among
the normal hypodermal cells. yp
Longitudinal sections show that in the intersegmental mem-
branes there is but a single layer of unmodified hypodermal cells
(fig. 8, i, and fig. 9, d). The nuclei of these cells are somewhat
elongate and flattened, with their long axes parallel to the surface
of the overlying cuticula; they are surrounded by but a small
amount of cytoplasm, so slight in cases that it appears thread-
like with the nuclei bulging out here and there. The nuclei
themselves are very deeply stained and are fairly regularly
arranged. THE JOURNAL OF MORPHOLOOY, VOL. 31, NO. 3 LOCATION AND STRUCTURE OF THE GLANDS 569 WAX GLANDS OF THE COCKROACH At the anterior end of each segment-that
part which is over-
lapped by the preceding segment-the modified hypodermal cells
are especially well developed, of ten producing the appearance
of two or more layers of cells, no doubt due to the crowding of
the individual cells, with the consequent displacement of their
nuclei (figs. 8 and 9). This is best seen on the ventral side of
the male where the cells are closely packed together so that there
seems to be no order to their arrangement. In such places the
cells also appear of different sizes, no doubt due to the fact that
a single section did not pass through the middle of each cell. In
other places they are but a single layer in thickness. p
y
g
y
These modified hypodermal cells possess large vesicular nuclei
with relatively thick nuclear walls, and provided with a nucleolus
and many deeply stained chromatin granules (fig. 10). Their
cytoplasm is also coarsely granular in appearance, and contains
several vacuoles. As a rule, they are cuboidal or columnar,
except where crowding together his caused them to assume vari-
ous forms. Scattered here and there between these cells and
the overlying cuticula, or intercalated somewhat between them
at their upper surface, are the non-glandular hypodermal cells
with nuclei similar to those in the intersegmental areas. Posterior to that portion of the segment which is overlapped
by the preceding segment, the normal hypodermal cells are in
the majority, while here and there scattered among them are
large modified cells similar to those occurring at the anterior end
of the segment. Where such cells occur, the ordinary hypoder-
mal cells are absent or else are somewhat crowded together. From the foregoing observations, therefore, it will be seen
that the body wall of the cockroach does not differ materially
from that of other insects, being composed of a cuticula, a hypo-
dermis consisting of a single layer of cells, and a basement mem-
brane. However, many of the hypodermal cells have become
specialized, increasing in size, so that in places, particularly on
the ventral side of the male, they present the appearance of two
or more layers. The cytoplasm of these cells has become strongly
granular while the nuclei have become large and vesicular. Their
entire structure, therefore, indicates that they are secretory in THE JOURNAL OF MORPHOLOOY, VOL. 31, NO. LOCATION AND STRUCTURE OF THE GLANDS 3 570 E. H. DUSHAM nature, and are therefore gland cells. Their secretion passes
to the exterior through very minute pores through the chitin
(fig. 7). These are evident only in very thin sections, cut ex-
actly perpendicular to the surface of the cuticula. Otherwise it
is almost impossible to make them out. With favorable sec-
tions they show as a fine streaking of the cuticula’ (fig. 10, p ) . nature, and are therefore gland cells. Their secretion passes
to the exterior through very minute pores through the chitin
(fig. 7). These are evident only in very thin sections, cut ex-
actly perpendicular to the surface of the cuticula. Otherwise it
is almost impossible to make them out. With favorable sec-
tions they show as a fine streaking of the cuticula’ (fig. 10, p ) . In portions of the cuticula stained black with osrnic acid the
pores also show as minute white dots under an oil-immerson lens. The secretion passes through these pores to the exterior and
without doubt spreads in a fine film over the outer surface of the
cuticula. y
g
g
In portions of the cuticula stained black with osrnic acid the
pores also show as minute white dots under an oil-immerson lens. The secretion passes through these pores to the exterior and
without doubt spreads in a fine film over the outer surface of the
cuticula. NATURE OF THE SECRETION Mingazzini, in his work on Periplaneta orientalis, stated also
that these modified cells were glandular in function and probably
secreted an oily substance. 80 much did these cells resemble,
both in appearance and position, those cells in Apis, Trigona,
Bombus, Melepona, and Aphrophora, which were definitely
known to secrete wax, that it occurred to the writer that possibly
they might also perform this same function in the case of the
cockroach. Tests were accordingly made to see if wax was pres-
ent in these insects. A large number of the insects were cap-
tured and thrown into a bottle of warmed chloroform. After
remaining in this liquid for ten minutes, they were strained out
before the chloroform could have had time to have penetrated
into the interior of the insect and attacked the fat in the fat
cells. The chloroform was then filtered and divided into two
portions. From one of these portions the chloroform was entirely
evaporated on a water-bath. When the chloroform hiid been
entirely driven off, a brownish liquid was left which possessed
the characteristic cockroach odor and which, on cooling, solidified
into a greasy appearing substance. The next point was to find out whether this subsfance was
grease or a wax. It is well-known that fatty oils, or fats treated
with basic hydroxides, are decomposed into fatty acids and glyc-
erol, this decomposition being hastened by using alcohol as a
solvent for the alkali. Both the fatty acids and glycerol are WAX GLANDS OF THE COCKROACH 571 soluble in water, so that when water is added no precipitate
would be formed. On the other hand, waxes, when similarly
treated, yield smaller amounts of fatty acids and, instead of
glycerol, give a large proportion of alcohol of the CnHancl series,
which is a solid body insoluble in water. Consequently, when
water is added to these, a distinct precipitate is formed. p
p
The greasy appearing material obtained from the cockroaches
was treated in this way. A small amount of pure beeswax was
similarly treated as a check. In both cases, after the two sub-
stances had been saponified and the liquid evaporated over st
water-bath, a decided precipitate was obtained on the addition of
water, more heavy in the case of beeswax, no doubt because
more of the wax was present. It seemed, therefore, that the
material secreted on the outside of the cockroach was wax. NATURE OF THE SECRETION In order to substantiate the preceding conclusion, a series of
solubility tests were made with the second portion of the chloro-
form in which the roacies had been dropped. These tests were
similar to those carried out by Hollande ('14) in his research on
the oenocytes of insects, when he demonstrated that the crystals
found in these bodies were wax crystals. In order to obtain wax
for these tests a few drops of the chloroform was placed on a
glass slide and then subjected to a current of air. By this means
the chloroform was quickly evaporated, leaving a small residue
of wax, appearing as a whitish spot on the slide. Slides thus
prepared were immersed in the various reagents for periods vary-
ing from fifteen minutes to one-half hour. Observations were
then made to see whether or not the material had been dissolved. Beeswax and wax from Pseudococcus citri dissolved in chloroform
were used as checks. The results are as per table, page 572. The saponification and solubility tests therefore clearly indi-
cate that wax is present on the body of the cockroach. There-
fore, because wax is @resent on the bodies of these insects and
glands are found in their abdomens resembling glands found in
other insects which are known to secrete wax, it seems conclu-
sive that these modified hypodermal cells are true wax glands. p
p g 572 E. H. DUSHAM _
_
REAGENT
-
. . . . . . . . . . . . . . . Alkaline soap. sol . . . . . . . . . . . . . . . Citric acid.. . . . . . . . . . . . . . . . Tartaric acid.. . . . . . . . . . . . . . Oxalie acid ................. Acetic acid.. . . . . . . . . . . . . . . . H2S04. . . . . . . . . . . . . . . . . . . . . . . HNO, ...................... HCL! ...................... Cold absol. alcohol.. . . . . . . . . . . . Acetone. . . . . . . . . . . . . . . . . . . . Olive oil .................... Cold alcoholic potash.. . . . . . . . . . NATURE OF THE SECRETION Ether . . . . . . . . . . . . . . . . . . . . . . Chloroform. . . . . . . . . . . . . . . . . Xylol ...................... Sodium carbonate . . . . . . . . . . . . . . Ammonium carbonate. . . . . . . . . . R E E I W A X
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Soluble
Soluble
Soluble
No coloration
Insoluble
Insoluble
COCKROACH
MATERIAL
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
rnsoluble
Soluble
Soluble
Soluble
No coloration
Insoluble
Insoluble
PS E UDOCOCCUS
W A X
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insoluble
Insqluble
Insoluble
Insoluble
Insoluble
Soluble
Soluble
Soluble
No coloration
Insoluble
Insoluble DEVELOPMENT OF THE GLANDS Mingazzini, in his work on Periplaneta orientalis, found that
these gland cells were less developed in the nymphs than in the
adult. In such immature stages he found that sections showed
only a single layer of cells, between which were observed fre-
quently large modified cells. In Blatta germanica these gland
cells were first distinguished during the first instar when the
nymphs are two days old (fig. 11). At that time some of the
normal hypodermal cells become modified. The nuclei which
are deeply stained and densely granular, gradually increase in
size still retaining their densely granular appearance. There\ is
also an increase in the amount of cytoplasm around these nuclei,
with the result that the adjacent normal hypodermal cells are
crowded so that their regular arrangement is somewhat broken. The growth in size of the nucleus and the increase of cytoplasm
also cause these cells to project below the level of the normal 573 WAX GLANDS OF THE COCKROACH hypodermis, so that they stand out rather conspicuously. Fi-
nally, the cytoplasm becomes granular, the dense chromatin gran-
ules in the nucleus seem to assemble in large clumps, and the
nucleolus, which up to this time could not be distinguished, be-
comes clearly visible. The transition from normal hypodermis
to glandular cells can thus be readily observed in two-day-old
nymphs. In the third instar the condition characteristic of the
adult is attained, the cells being fully developed, and in some
cases so closely packed $ogether as to simulate the appearance
of two or more layers. BIBLIOGRAPHY ALLEN 1910 Commercial organic analysis, vol. 2. BERLESE, A. 1909 Gli Insetti, pp. 470-471, 496-502 pp
BUGNION, M. E. 1908 Glandes cirieres de Flata (Phromia) marginella (Ful-
goridae). Archiv des Sciences Physiques et Naturelles, T. 25. 9 CARLET, G. g
1890 Sur les organes secreteurs et la secretion de la cire chez
l’abeille. Comptes Rendus de 1’Academie des Sciences, Paris, T. 110. DEEGENER,
P. 1912 Handbuch der Entomologie, S. 37-42. g
DREYLING,
L. 1903 Vber die wachsbereitenden Organe der Honigbiene. Zool. Anz., Bd. 26. 1903 Weitere Mitteilungen uber die wachsbereitenden Organe der
Honigbiene. Zool. Anz., Bd. 27. 1905 Beobachtungen uber die wachsbereitenden Organe bei den Hum-
meln, nebst Bemerkungen uber die homologen Organe bei Trigonen. Zool. Anz., Bd. 29. 1905 Die wachsbereitenden Organe bei den geselliglebenden Bienen. Zool. Jahrb., Bd. 22. 1903 Weitere Mitteilungen uber die wachsbereitenden Organe der
Honigbiene. Zool. Anz., Bd. 27. 1905 Die wachsbereitenden Organe bei den geselliglebenden Bienen. Zool. Jahrb., Bd. 22. DUJARDIN, M. F. 1849 MBmoire sur 1’6tude microscopique de la cire. Annales
des Sciences Naturelles, T. 12; 3d ser. GUILLBEAU, B. H. 1908 The origin of the froth in spittle insects. Amer. Nat.,
vol 42. HOLLANDE,
A. 1914 Les cerodecytes ou oenocytes des insectes consideres ail
point de vue biochimique. Archiv d’Anat. Micr. p
q
MAYER, P. 1891-93 Zur Kentniss von Coccus cacti. Mittheil. Zoolog. Station
zu Neapel, Bd. 10. p
METALINKOFF, S. 1904 Sur un procede nouveau pour faire des coups micro-
scopiques dans les animaux pourvu d’un tegument chitineux epais. Archiv Zool. Exp.; Notes et Revue, T. 4, ser. 2. p ;
,
,
MIALL, L. C., AND DENNY,
A. 1886 The structure and life history of the cock-
roach. MINCHIN, E. A. 1888 Note on a new organ and on the structure of the hypoder-
mis in Periplaneta orientalis. Jour. of Micr. Sci., N. S., vol. 29. p
MINGAZZINI, P. 1889 Recerche sulla struttura dell’ipodermide nella Peri-
planeta orientalis. Atti della Reale Accademia dei Lincei, vol. 5,
ser. 4. NUSSLIN, 0. 1900 Zur Biologie der Schizoneuriden Gattung Mindarus Koch. Biol. Centralblatt, Bd. 20. OETTINGER, 0. 1906 Uber die Drusentaschen am Abdomen von Periplaneta
orientalis und Phyllodromia germanica. Zool. Anz., Bd. 30. y
g
PACKARD,
A. S. 1898 Text-book of Entomology, pp. 361-366. SUNDWICK,
E. E. 1907 uber das Wachs der Hummeln. Zeit. f. Physiol. Chemie,
Bd. 53. VIALLANES, H. 1882 Histologie et developpement des Insectes. Annales des
Sciences Naturelles, T. FUNCTION OF THE WAX The secretion of wax in insects serves various purposes-pro-
tection against enemies, cold, and moisture, a protection for
eggs, an encasement for excreta, a lining for the larval burrow,
etc. In the cockroach it probably serves as a protection, against
moisture, for they live in protected places where other enemies
are unable to attack them. On the other hand, their presence in
warm, damp places intimately associated with water would seem
to indicate that the wax was secreted to protect them from this. p
The fact that the wax possesses the characteristic ‘roachy’
odor suggested the idea that it might be a protection against
enemies on account of its smell. However, an examination of
beeswax and Pseudococcus wax showed that these also possessed
characteristic odors. That of beeswax is well known. The
odor from Pseudococcus was like mouldy wet leaves. That of
the cockroach has already been described. From this it would
seem that the wax secreted by each different family of insects had
an odor ‘sui generis.’ The fact that the ‘roachy’ odor remains
in roach-infested places for a considerable period of time after
the roaches have been driven away or exterminated and that it
is difficult to remove it from dishes and kitchen utensils may be
accounted for by the fact that the wax remained adhering to
these things over which they were accustomed to run. g
y
The writer wishes to acknowledge his sincere thanks.to Dr. W. A. Riley, whose advice and criticism have been invaluable
and whose encouragement has made this work possible. 574 E. H. DUSHAM BIBLIOGRAPHY 14, ser. 6. WITLACZIL, E. 1885 Die Anatomie der Psylliden. Zeit. f. wiss. Zool., vol. 42. ,
, PLATES 575 EXPLANATION OF FIGURES g
g
2 Ventral abdominal surface of male, showing distribution of the wax glands. 576 576 WAX GLANDS OF THE COCKROACH
E. H. DUSHAM WAX GLANDS OF THE COCKROACH
E. H. DUSHAM PLATE 1 PLATE 1 WAX GLANDS OF THE COCKROACH
E. H. DUSHAM 2
577 2 2 2 2 577 EXPLANATION O F FIGURES 3 Dorsal abdominal surface of female, showing distribution of the wax
glands. 4 Ventral abdominal surface of female, showing distribution of the wax
glands. 578 578 578 PLATE a PLATE a WAX GLANDS OF THE COCKROACH
E. H. DUBHAM 579 EXPLANATION OF FIGURES 5 Surface view of the wax glands (focus below the non-glandular hypoder-
mis). X 600. 5 Surface view of the wax glands (focus below the non-glandular hypoder-
mis). X 600. )
6 Surface view of normal hypodermis of the intersegmental membrane. X
WO. 7 Surface view of cuticula overlying the glandular areas showing pores. X
600. 8 Longitudinal section through a single intersegmental area on the dorsal
surface of the female, lateral to the middle line, showing the appearance and
location of the wax glands. i, intersegmental membrane; c, cuticula; h, hypoder-
mis; g, wax glands; b, basement membrane. X 160. g
g
,
9 Longitutinal section through a single intersegmental area on the ventral
surface of the male, showing appearance and location of the wax glands. a,
muscles; c, wax glands, b, basement membrane; d, intersegmental membrane;
e, hypodermis; j, cuticula. X 160. ,
yp
;j,
10 Portion of the body wall in the region of the wax glands, showing detailed
structure. n, non-glandular hypodermis; p , pores; c, cuticula; v, vacuole; T,
basement membrane. (No. 10 ocular, 1.8-mm. oil immersion.) (
11 Longitudinal section through a single intersegmental area of a two-day-
old nymph, showing the development of the gland. u, hypodermis; t, transform-
ing hypodermal cell; s, fully-transformed cell. X 600. 580 WAX GLANDS OF THE COCKROACH
E. H. DUEHAM WAX GLANDS OF THE COCKROACH
E. H. DUEHAM PLATE 3 PLATE 3 5 7 7 6 7 5 8 8 8 8 9 9 9 P
10
11
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Tenemos a una mujer que se apropia del relato construyendo su propio discurso mediante su acción política basada en la solidaridad como
principio ideológico, pues como indica: «Nosotros aquí comiendo y bailando tranquilos, mientras otros compañeros, como nuestro pata Plasencia, el motorista de Pescamar, agonizan en el hospital, o están recluidos
en las cárceles, o los policías los tienen haciéndoles botar piedras en las
pistas a punta de metralleta» (Colchado, 2011, p. 458). Ese discurso tiene un lugar de discusión y enunciación, el bar y unos interlocutores que
difícilmente la discursividad histórica oficial validaría como fuentes. Aquí,
el sentido político del discurso radica en la vivencia de la desigualdad y en
práctica política a la que conduce.
25 El escritor chimbotano Augusto Rubio Acosta (2007), en su crónica «De cómo alcanzar la
libertad», dedicada a la activista social, afirma: «Ojalá la mayoría de mujeres chimbotanas
tuvieran el coraje de enfrentarse a su destino —pensamos—, ojalá tuvieran la sensibilidad
social y la necesidad de libertad a flor de piel. Ojalá hubiera Saras en cada empresa, en cada
barrio, en cada comunidad, en cada ministerio, para hacer del país un Perú distinto».
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Intención historiográfica y sujetos de relato en Hombres de mar, de Óscar Colchado Lucio
Hacia el final de la novela, el devenir histórico social alcanzará hasta
entrado el 2000: la Marcha de los Cuatro Suyos, la huida de Fujimori y su
insólita y vergonzosa renuncia a la presidencia desde Japón mediante
un fax. Precisamente, cuando se prepara la Marcha de los Cuatro Suyos,
vemos que el discurso de Sarandonga se ha transformado en memoria
colectiva, como una conciencia crítica de la realidad que la coloca en un
lugar de respeto absoluto. Así lo explica un esporádico narrador:
Una señora delgada, de tez clara y cabellos largos, que yo vi arengando a la multitud, pasó corriendo por mi lado, asustada, volviendo
el rostro de cuando en cuando para constatar si los que la seguían
estaban próximos. Dijeron que se trataba de la luchadora social Sara
Sarandonga que huía de los guardias que la venían persiguiendo en
un carro. Yo la había visto ya otras veces presidiendo las marchas en
las huelgas, mas nunca como hoy en apuros (496).
El discurso de Sara es su propia práctica política, porque el bien común va más allá de la palabra; para ella, la política sigue siendo vivencia y
cuerpo. Así nos explica que cuando «un grupo de pescadores que la escuchaba atento, un guardia me vino a amenazar con su revólver. Tú ya eres
muerta para mí, me dijo. Yo no me chupé, compañeros. ¡Dispara, pue, baboso!, le dije, ¡dispara! Y entre mí dije: Qué me importa, muero yo o muere
él, y me lancé a quitarle el arma» (Colchado, 2011, p. 496).
Tenemos pues en el texto algo más que personajes. La voz de Sara
Sarandonga es constantemente validada por su estatuto testimonial que,
como se ha dicho, ha sido recuperado y documentado por el autor. Ella
relee desde los márgenes del discurso social los discursos de poder, los
mecanismos y modulaciones de la violencia política, estatal, económica
y racial, pero también, en cuanto sujeto de relato, el discurso de Sarandonga es una respuesta y es acción. Se trata, finalmente, de validar y reconocer en sujetos como Sarandonga las nuevas formaciones sociales y
culturales obviadas, muchas veces, por el discurso historiográfico de cuño
academicista.
Algunas conclusiones
La naturaleza problemática de la bien llamada «monumental novela»
de Colchado, Hombres de mar, está dada en gran medida por la diversidad de voces intencionadas historiográficamente, que se imbrican en la
obra, poniendo en la superficie textual diversas versiones de la historia y,
por consiguiente, diversas capas de realidad y memorias, muchas veces
diametralmente opuestas. La estructura polifónica de Hombres de mar,
más que activar memorias de consenso a través de las voces de sujetos
de relato, moviliza una pluralidad de memorias sociales disputando el
Desde el Sur | Volumen 9, Número 2
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Estefanía Peña Steel
derecho a contar cada una su propia verdad o versión del hecho experimentado. En este sentido, el texto en su heterogeneidad sociocultural
llega a ser esa «zona de ambigüedad y conflicto», de la que nos habla Cornejo Polar al definir las literaturas heterogéneas. En escrituras éticas como
las de Arguedas o Colchado el sujeto que enuncia sí importa. Con todo, no
se trata aquí de negar la objetividad factual de la Historia en cuanto disciplina, sino de reconocer las fisuras de su enunciación univoca por donde
se infiltran aquellas voces desautorizadas por la oficialidad y que en su acción extra e intra textual, nos recuerdan lo concreto del «hecho literario».
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Referencias bibliográficas
Angenot, M. (2010). El discurso social. Los límites de lo pensable y lo decible.
Buenos Aires: Siglo XXI.
Arguedas, J. M. (1983). El zorro de arriba y el zorro de abajo. Lima: Horizonte.
Birulés, F. (1989). Introducción. En A. Danto. Historia y narración (pp.
9–27). Barcelona: Paidós.
Bueno, R. (2004). Sobre la heterogeneidad literaria y cultural de América
Latina. En Antonio Cornejo Polar y los avatares de la cultura latinoamericana
(pp.19–36). Lima: Fondo Editorial de la Universidad Nacional Mayor de San
Marcos.
Castro, D. (2005). Una reflexión. En El encuentro peruano de narradores
en Madrid. Omnibus. Recuperado de http://www.omnibus.com/congreso/
opiniones/dantecastro.html
Chartier, R. (1999). Trabajar con Foucault: esbozo de una genealogía de
la «función-autor». Signos Históricos, 1, pp. 11–27.
Colchado, Ó. (2011). Hombres de mar. Lima: Alfaguara.
Cornejo, A. (1989). Los sistemas literarios como categorías históricas.
Elementos para una discusión. Revista de Crítica Literaria Latinoamericana,
29, pp. 19–25.
_______________ (julio-diciembre de 1996). Una heterogeneidad no
dialéctica: Sujeto y discurso migrantes en el Perú moderno. Revista Iberoamericana, LXII(176–177), pp. 837–844.
Danto, A. (1989). Historia y narración. Ensayos de filosofía analítica de la
historia. Barcelona: Paidós.
Degregori, C. I. (2007). Entre el fuego y la calandria. Visión del Perú desde la narrativa andina. Crónicas urbanas, 12, pp. 55–66.
Even-Zohar, I. (1999). Factores y dependencias de la cultura. Una revisión de la teoría de los polisistemas. Teoría de los polisistemas. Madrid:
Arco.
Foucault, M. (1999). ¿Qué es un autor? En Entre filosofía y literatura.
Obras esenciales. Volumen I. Barcelona: Paidós.
Grez, S. (2005). Historiografía, memoria y política. Observaciones para un
debate. Recuperado de http://web.uchile.cl/vignette/cyberhumanitatis/
CDA/texto_simple2/0,1255,SCID%253D21039%2526ISID%253D730,00.
html
Guattari, F. y Rolnik, S. (2006). Micropolítica. Cartografías del deseo. Madrid: Traficantes de Sueños.
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Harshaw, B. (1997). Ficcionalidad y campos de referencia. En G. Antonio.
Teorías de la ficción literaria (pp.126–158). Madrid: Arco Libros.
Huamán, M. Á. (2006). Escritura utópica y crítica estético-política: de Churata a Colchado. Escritura y Pensamiento, 19, pp. 7–22.
Miranda, P. (2014). Moncada, el profeta de la ecología. Chimbote: Río Santa Editores.
Matos-Mar, J. (1986). Desborde popular y crisis del Estado. El nuevo rostro
del Perú en la década de 1980. Lima: Instituto de Estudios Peruanos.
Montoya, R. (1978). El carácter predominantemente capitalista de la economía peruana actual. (1960–1979). Lima: Mosca Azul Editores.
Ortega, J. (2010). El sujeto dialógico. Negociaciones de la modernidad conflictiva. Ciudad de México: Fondo de Cultura Económica.
Osorio, J. (1995). La narrativa andina. Sieteculebras, 8, pp. 9–10.
Pazos, J. P. (2004). Realidad y ficción: las huellas del «Loco Moncada» en
el imaginario popular a fines de los noventa en Chimbote. En W. Kapsoli
(Comp.). Zorros al fin del milenio (pp. 87–110). Lima: Centro de investigación Universidad Ricardo Palma.
White, H. (1992). Metahistoria, la imaginación histórica en la Europa del siglo XIX. Ciudad de México: Fondo de Cultura Económica.
_______________ (2003). El texto histórico como artefacto literario y otros
escritos. Barcelona: Paidós.
Recibido: febrero de 2017
Aprobado: abril de 2017
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Rezensionen
Seemacht. Eine Seekriegsgeschichte von der Antike bis zur Gegenwart. Herausgegeben
von Elmar B. Potter und Chester W. Nimitz. Deutsche Fassung herausgegeben von Jürgen Rohwer. München: Bernard & Graefe 1974. X V I I I , 1201 Seiten
Dieser umfangreiche Band verdankt sein Entstehen dem Wunsch des damaligen Inspekteurs der
Bundesmarine und späteren Präses des Arbeitskreises für Wehrforschung, Vizeadmiral a . D.
Professor Friedrich Ruge, ein auf dem heutigen Stand der Forschung stehendes Handbuch zu
Fragen der Seemacht und Seekriegsgeschichte vorzulegen. Laut Vorwort schien jedoch eine
selbständige deutsche Veröffentlichung nicht möglich, so daß auf ein im Jahre 1960 erschienenes
amerikanisches Handbuch 1 als Grundlage zurückgegriffen wurde. Die von zwölf Mitarbeitern
geschriebenen Kapitel des Originals wurden von neun Bearbeitern übersetzt, teilweise gekürzt
oder erweitert und durch neue Beiträge ergänzt: deshalb zählt die deutsche Ausgabe nun 52 Kapitel. Dabei wurde die Darstellung bis zum Jahre 1973 geführt, ein neues Literaturverzeichnis
erstellt, das stärker die deutschsprachige Forschung berücksichtigt, sowie ein Personen- und ein
Schiffsverzeichnis angelegt. Auf Anmerkungen ist dagegen fast völlig verzichtet worden.
Zur Konzeption des Werkes erläutern die amerikanischen Herausgeber, daß sie weitgehend die
Analyse der Seekriegsgeschichte von Alfred T . Mahan 2 übernommen und auf die späteren
Ereignisse angewandt hätten. Es sei »die Pflicht jeder an den Problemen der nationalen Verteidigung interessierten Person, ein solches Buch zu lesen und zu verstehen«. Das Hauptinteresse
liegt - auch in der deutschen Ausgabe - bei der jüngeren Geschichte. Für die Zeit von der Antike
bis zum amerikanischen Unabhängigkeitskrieg werden 70 Seiten benötigt, bis zum Zeitalter des
Imperialismus weitere 185. Die Entwicklung bis zum Ende des Ersten Weltkrieges wird dann
auf fast 200 Seiten geschildert und der Zweite Weltkrieg sogar auf über 400.
Bei der Vielzahl der Mitarbeiter ist es trotz aller Abstimmung wohl nicht zu vermeiden gewesen,
daß die einzelnen Kapitel sehr unterschiedlich ausgefallen sind. Die Geschichte der Seefahrt in
der Nord- und Ostsee von der mittleren Steinzeit bis zum Ende des 16. Jahrhunderts sowie der
Züge von Germanen, Wikingern und Normannen läßt sich auf achteinhalb Textseiten kaum befriedigend darstellen; das gleiche gilt für das Zeitalter der Entdeckungen und für die deutschen
Flotten im 17. und 18. Jahrhundert. Die Bezeichnung »Flotten« wird hier auch ziemlich großzügig gebraucht, denn der Verfasser spricht schon bei der Indiensthaltung von zwei Wachfahrzeugen davon, daß ein Staat Seemacht entwickelte. - In zwei Kapiteln über Politik und Flottenbau und über die technische Entwicklung in den Marinen in den Jahren vor 1914 betont Jürgen
Rohwer den Einfluß der Technik auf den Flottenbau und führt damit Gedanken fort, die er bereits auf der Tagung »Marine und Marinepolitik im kaiserlichen Deutschland 1871-1914« vorgetragen hat 3 . Hier wäre allerdings zu wünschen, daß die vor allem von Volker R. Berghahn
ausgelöste Kontroverse um die Ziele des Tirpitzschen Flottenbaues einen stärkeren Niederschlag gefunden hätte 4.
Ein weniger gut gelungenes Kapitel ist leider der Handelskrieg im Ersten Weltkrieg. Der amerikanische Autor hat offenbar fast nur die bis 1934 erschienene Literatur gekannt und auch der
deutsche Bearbeiter hat die in der Bibliographie aufgeführten jüngeren Arbeiten nicht benutzt.
Die deutsche Kriegsgebietserklärung vom 4. Februar 1915 wird von ihm aus dem Englischen
rückübersetzt, statt den deutschen Originaltext bei Spindler 5 heranzuziehen : dann hätte er auch
bemerkt, daß die Erklärung in Bd 1 und nicht in Bd 3 abgedruckt ist. In der amerikanischen
Ausgabe vorhandene Anmerkungen werden gestrichen, ohne daß die Zählung berichtigt wird,
und eine Anmerkung ist im Text so verrutscht, daß ihr Bezug nicht mehr zu erkennen ist. In
zwei Anmerkungen wird auf Arbeiten aus den 50er Jahren verwiesen, die aber nicht im Literaturverzeichnis erscheinen. Die Angaben über die Konstruktionsdaten der deutschen U-Boote
sind äußerst ungenau, und die Zahlen über Stärke, Verluste, Neuzugänge und Erfolge der U Boot-Waffe häufig falsch. Bei der Darstellung der deutschen U-Boot-Offensiven wird ausgeführt, daß nach dem »Sussex«-Zwischenfall im März 1916 der Handelskrieg um England eingestellt wurde, bis der Kaiser am 9. Januar 1917 den Beginn des uneingeschränkten U-Boot-Krieges zum 1. Februar befahl. Bei der Schilderung der britischen Abwehr vier Seiten weiter wird
von erfolgreichen U-Boot-Operationen im September 1916 im Kanal berichtet und von monatlichen Verlusten, die schon 150 Schiffe betrugen - eine Klärung des Widerspruchs erfolgt nicht.
127
M G M 2/75
Im Bereich der Dover-Sperre wurden zwischen November 1917 und September 1918 zwölf U-
Boote vernichtet: hier geschieht dies »um den August 1918 herum«, und dadurch wird angeblich
sogar die Aufgabe der U-Boot-Basis Flandern erzwungen. Andererseits wird behauptet, die Alliierten hätten die »Northern Barrage« nicht gegen Räumversuche sichern können, und Minensuchboote hätten dort die deutschen U-Boote begleitet. Völlig verfehlt ist auch in der Zusammenfassung der Versuch, einen direkten Zusammenhang zwischen der Einführung des Geleitsystems und den steigenden U-Boot-Verlusten herzustellen.
Wesentlich besser ist dagegen das Kapitel über die Schlacht im Atlantik 1939-1945 gelungen.
Hier konnte Jürgen Rohwer auf eigene Arbeiten zurückgreifen 6 und das amerikanische Original berichtigen und ergänzen. Ähnliche Erweiterungen erfuhren auch andere Kapitel: so wurden bei den Überwasseroperationen im Atlantik allein neun Skizzen für die Operation »Rheinübung« neu aufgenommen.
Bei der Darstellung des Krieges im Pazifik wurde eine gekürzte Fassung zugrunde gelegt. H i e r wie auch im Original - wird häufig eine genaue Nennung der an den Operationen beteiligten
Verbände und Einheiten vermißt, sowie Ubersichten, die einen besseren Eindruck von den beiderseitig zur Verfügung stehenden Kräften vermitteln.
Die Beiträge, die sich mit den Ereignissen nach 1945 befassen, sprengen dagegen den Rahmen
einer Seekriegsgeschichte, wie er von Edward Wegener im letzten Kapitel über die Rolle der
Seemacht in unserer Zeit beschrieben wird: »Die Unterstützung des Kampfes zu Lande gehört
in seinen Einzelheiten allerdings nicht in den Bereich der Seekriegführung.« Vor dem Hintergrund des Kalten Krieges der Nachkriegszeit über die Strategie der massiven Vergeltung zur abgestuften Abschreckung werden die Konflikte der jüngsten Zeit geschildert, wobei besonders
dem Korea-Krieg und dem Vietnam-Krieg breiter Raum gewidmet wird. Aber auch die Kriege
wie die zwischen Israelis und Arabern und zwischen Indern und Pakistanis werden behandelt.
Hier zeigt sich wieder einmal die Problematik, in die der Historiker geraten kann, wenn er sich
um Aktualität bemüht. So wird das Urteil über den Waffenstillstand in Vietnam vom Januar
1973 heute kaum noch aufrechterhalten werden: »Damit hatte Präsident Nixon sein Ziel, die
USA aus dem Vietnamkrieg zu lösen, ohne den Verbündeten preisgeben zu müssen, erreicht.«
Bernd Stegemann
Anmerkungen
1
2
3
4
5
6
Seapower. A navel history. Ed.: E. Β. Potter, Ch. W. Nimitz. Englewood Cliffs, Ν . J. 1960.
A. T. Mahan: The Influence of sea power upon history, 1660-1783. Boston 1890; ders.: The Influence
of sea power on the wars of the French Revolution and Empire. Vol. 1.2. Boston 1901. Vgl. jüngst P. M.
Kennedy: Mahan versus Mackinder. Two interpretations of British sea power. In: M G M 16 (1974)
39-66.
J. Rohwer: Kriegsschiffbau und Flottengesetze um die Jahrhundertwende. In: Marine und Marinepolitik im kaiserlichen Deutschland 1871-1914. Hrsg. von H . Schottelius und W. Deist. Düsseldorf 1972,
S. 211-235.
V. R. Berghahn: Zu den Zielen des deutschen Flottenbaus unter Wilhelm II. In: H Z 210 (1970) 34-100;
ders.: Der Tirpitz-Plan. Genesis und Verfall einer innenpolitischen Krisenstrategie unter Wilhelm II.
Düsseldorf 1971 ( = Geschichtliche Studien zu Politik und Gesellschaft. Bd 1.); ders.: Rüstung und
Machtpolitik. Zur Anatomie des »Kalten Krieges« vor 1914. Düsseldorf 1973 ( = Mannheimer Schriften
zur Politik und Zeitgeschichte. 5.). Zu beiden letzten die Bespr. in M G M 11 (1972) 196-198 und ebd. 15
(1974) 278 f.
A. Spindler: Der Handelskrieg mit U-Booten. Bd 1-5. Berlin, Frankfurt a. M. 1932-66.
J. Rohwer: DerU-Bootkrieg und sein Zusammenbruch 1943. In: Entscheidungsschlachten des Zweiten
Weltkrieges. Hrsg. von H . - A . Jacobsen und J. Rohwer. Frankfurt a.M. 1960, S. 327-394; ders.: Die
größte Geleitzugschlacht des Krieges. H X . 229/SC. 122 (März 1943). In: WWR 18 (1968) 146-158.
Documenta Bohémica Bellum tricennale illustrantia. Hauptredaktion: Josef Koci, Josef
Polisensky, Gabriela Cechová. T. 1.2. Praha: Academia, nakladatelstivi Ceskoslovenské
akademie vëd. Wien, Köln, Graz: Böhlau 1971-72.
1. Josef Polisensky: Der Krieg und die Gesellschaft in Europa 1618-1648. Aus dem
Tschechischen von Anna Urbanová. 1971. 237 Seiten
2. Der Kampf um Böhmen, Quellen zur Geschichte des Böhmischen Krieges
<1618—1621>. Herausgegeben von Miroslav Toegel. Aus dem Tschechischen von Anna
Urbanová. 1972. 338 Seiten
Mit den beiden vorliegenden Bänden ist der erste Schritt zu einer groß angelegten Edition von
Archivalien über den Dreißigjährigen Krieg getan, die sich in Archiven und Bibliotheken der
Tschechoslowakei befinden, bis heute jedoch noch nicht in zufriedenstellender Form erschlossen worden sind und daher für die Beurteilung des Krieges nicht benutzt werden konnten. Von
besonderem Interesse sind die im Zuge der nach 1945 vorgenommenen Verstaatlichung des
Großgrundbesitzes in Staatsbesitz übergegangenen Archive der großen Familien des Landes,
deren Mitglieder während des Dreißigjährigen Krieges eine bedeutende politische oder militärische Rolle gespielt haben. Genannt seien hier die Namen Slavata, Piccolomini, Gallas, Colloredo-Mansfeld, Lobkovic, Waldstein, Kolovrat, Buquoy, Verdugo, Schwarzenberg und Dietrichstein. Damit steht zusammen mit den staatlichen und städtischen Archivalien ein Material
zur Verfügung, das in seiner Geschlossenheit seinesgleichen sucht.
Der 1. Band, der die Einleitung zu dem Quellenwerk darstellt, ist von Josef Polisensky verfaßt
worden, einem Kenner des Dreißigjährigen Krieges, der jedoch, das sei vorweggenommen, sich
der marxistischen Geschichtsauffassung verpflichtet fühlt und daher entsprechend beurteilt und
kommentiert und sicher nicht in allem die ungeteilte Zustimmung nichtmarxistischer Historiker
finden wird.
Schon die Gliederung in die drei Hauptteile:
A. Zweck und Grundsätze der Edition von Quellen aus tschechoslowakischen Archiven,
B. Probleme und Quellen zur Geschichte des Dreißigjährigen Krieges,
C . Vorläufige methodologische Schlußfolgerungen läßt erkennen, daß man es nicht mit einer
Einleitung im herkömmlichen Sinne zu tun hat. Auch wenn der Verfasser es ausdrücklich bestreitet, entsteht doch hier und da der Eindruck, daß er mit dieser Einleitung so etwas wie eine
Gebrauchsanweisung für die weitere Forschung geben möchte.
Die Ausführungen Polisenskys sind derart breit angelegt und berühren eine so beachtliche Fülle
von Fragen und Problemen, daß hier nur auf die wesentlichsten eingegangen werden kann.
Polisensky geht davon aus, daß manches Ergebnis der traditionellen Geschichtsschreibung, die
mit Mythen, Legenden und Schemata gearbeitet habe, zu revidieren sei und daß der Dreißigjährige Krieg in verstärktem Maße vor dem Hintergrund der sich seit dem 16. Jahrhundert verändernden europäischen Gesellschaft zu sehen sei. Seine Aufmerksamkeit schenkt er weniger den
konfessionellen Veränderungen und Gegensätzlichkeiten als vielmehr den auftretenden revolutionären Bewegungen - ζ. Β. in England und den N i e d e r l a n d e n - u n d den gesellschaftlichen Gegebenheiten überhaupt. Daraus ergibt sich für ihn die Notwendigkeit einer neuen Periodisierung des Krieges.
Der zweite Teil, der eigentliche Hauptteil des Bandes, bringt eine Auseinandersetzung mit den
Problemen und Quellen zur Geschichte des Krieges; jedes der sieben Kapitel ist jeweils in drei
Abschnitte unterteilt, die
1. den gegenwärtigen Stand der Kenntnisse nach der neuen Literatur - letztlich aus marxistischer
Sicht - skizzieren,
2. eine Charakteristik der wichtigsten Quellen und Archivbestände anführen und
3. in praktischer Weise auf ein bestimmtes Problem hinweisen, das auf Grund der fraglichen
Quellen geklärt werden kann.
Nach der Auseinandersetzung mit den gegenwärtigen Versuchen einer neuen Auffassung des
Konflikts folgt in den weiteren sechs Kapiteln die Betrachtung der einzelnen Perioden des Krieges, wie Polisensky sie sieht:
1. Die Problematik des Böhmischen Krieges 1618-1621,
2. Der niederländische Konflikt 1621-1625,
3. Der dänische Eingriff und die Bemühungen um die Bildung der Großen Koalition 1625-1630,
4. Der schwedisch-niederländische Konflikt 1630-1635,
5. Der schwedisch-französische Konflikt 1635-1643,
6. Krieg, Revolution und Friedensverhandlungen 1644-1650.
Jeder dieser Perioden soll ein geschlossener Quellenband entsprechen.
Im Schlußteil geht Polisensky auf die Kriterien der Auswahl der Quellen - eine Auswahl mußte
in Anbetracht des umfangreichen Materials erfolgen - und die Art ihrer Veröffentlichung ein.
Sicherlich läßt es sich nicht vermeiden, daß mit jeder Auswahl zwangsläufig eine Wertung einhergeht, doch möchte man hoffen, daß sie hier nicht in der bestimmmten Absicht erfolgte, eine
vorgefaßte ideologisch motivierte Meinung über die »gesellschaftlichen« Hintergründe des
Dreißigjährigen Krieges zu untermauern.
Ein ausführliches Personen- und Ortsnamenregister, in das auch die Namen der im Text behandelten Autoren eingearbeitet sind, schließen den Einleitungsband ab, der an manchen Stellen sicherlich eine Straffung vertragen hätte. Die Übersetzung aus dem Tschechischen ist durchweg
glatt. Störend für den deutschsprachigen Leser, für den ja diese Übersetzung eigens angefertigt
wurde, ist jedoch die Tatsache, daß die Ortsnamen, sofern es sich nicht um historische Schauplätze handelt, in der slawischen Form erscheinen (z.B. Cheb statt Eger, Libérée statt Reichenberg, Bratislava statt Preßburg), doch geht dieses Manko nicht zu Lasten der Übersetzerin.
Der 2. Band, den Miroslav Toegel herausgegeben hat, bringt ausgewählte Quellen zur Geschichte des Böhmischen Krieges. Dazu sind die Bestände des Zentralstaatsarchivs in Prag, der
Staatsarchive in Brünn, Wittingau, Leitmeritz, Prag, Troppau, Pilsen und Zámrsk sowie die
Handschriftenabteilung des Nationalmuseums Prag durchgesehen worden. Aus der Fülle des
Materials wurden für die Zeit von 1617 bis 1621 über 900 Dokumente ausgewählt und im Volltext oder in Form von Kurzregesten und Auszügen veröffentlicht; beachtlich ist dabei die Zahl
bisher nicht bekannter Quellen.
Die Einleitung zu diesem Quellenband stammt von Josef Polisensky, der davon überzeugt ist,
daß der Ausbruch des Böhmischen Krieges auf den Verfall der Gesellschaft, insbesondere des
böhmischen Adels, zurückzuführen sei. Es steht für ihn fest, daß der Böhmische Krieg, der den
Dreißigjährigen Krieg einleitete, von Anfang an »ein Konflikt von kontinentalen Ausmaßen
war«.
Sicherlich wird das veröffentlichte Quellenmaterial aus den Archiven der Tschechoslowakei in
Verbindung mit den Ausführungen Polisenskys dazu beitragen, manche Frage zu überdenken
und unter einem neuen Aspekt zu sehen. Vielleicht wird es dann möglich sein, zu neuen, genaueren Kenntnissen über den Dreißigjährigen Krieg zu gelangen.
Peter Veddeler
Peter-Christoph Storm: Der Schwäbische Kreis als Feldherr. Untersuchungen zur Wehrverfassung des Schwäbischen Reichskreises in der Zeit von 1648 bis 1732. Berlin : Dunkker & Humblot 1974. 597 Seiten ( = Schriften zur Verfassungsgeschichte. Bd 21.)
Nach einem Wort Fritz Hartungs rechnete die Geschichte der Reichskreise lange Zeit »zu den
am wenigsten erforschten Gebieten der Reichsverfassung«1 und - wie wohl zu ergänzen ist darüber hinaus der politischen Wirklichkeit im sacrum imperium. Galt Hartungs Aussage für
die Konstitution uneingeschränkt, so betraf das letztere hauptsächlich die Lande am Rhein,
Main und Neckar; denn hier hatte das Reich nicht zuletzt dank der Kreise, in denen sich eine
Vielzahl mindermächtiger geistlicher und weltlicher Fürsten, Prälaten, Grafen, Herren und
Städte zur Kooperation zusammenfand, noch bis an die Schwelle des 19. Jahrhunderts ein nicht
zu unterschätzendes Gewicht. Die historische Forschung hat sich indes fast ausschließlich den
aufstrebenden Landesstaaten zugewandt, vor allem den Vormächten Österreich und Brandenburg-Preußen und daneben den größeren Territorien Bayern, Braunschweig-Lüneburg, Sachsen, Württemberg. Hingegen wurde das Reich, über das nach dem Westfälischen Frieden höchstens von Stagnation, Verfall und Auflösung zu berichten war, nur allzugerne als beinahe bedeutungslos abgetan; folglich fanden interterritoriale Korporationen wie die Reichskreise oder die
Ritterkreise und -kantone erst recht keine Beachtung. In der jüngsten Vergangenheit jedoch,
seit die Forderung nach europäischer Integration auch das Interesse an früheren Föderativsystemen geweckt hat, hat die Zahl historischer Studien zugenommen, die sich mit internationalen
und interterritorialen Zusammenschlüssen, ihren Institutionen und Formen der Zusammenarbeit sowie ihrer politischen Wirksamkeit und ihrem Einfluß befassen. Diesem Trend folgt Storm
in seiner Tübinger juristischen Dissertation, wobei man ihm aber bescheinigen muß, mit seiner
breit angelegten und materialreichen Untersuchung für den Schwäbischen Kreis weitgehend
Neuland bearbeitet zu haben.
Anliegen des Autors ist es, die Wehrverfassung des Schwäbischen Kreises, »einer intermediären
Verfassungseinrichtung zwischen Reich und Stand« (S. 40), für einen Zeitraum von knapp 100
Jahren zu analysieren, in denen der Schritt von der Exekution zur Armatur vollzogen wurde
und dieser Kreis mit dem Aufbau eines Stehenden Heeres - das nach wie vor in erster Linie ein
Teil des Reichsheeres war - neben die sogenannten armierten Stände trat, ja, zeitweise zusammen mit einigen benachbarten Kreisen sogar zum Partner europäischer Mächte wurde. Tatsächlich sollte es diese Kooperation den rund 100 schwäbischen Fürsten und Ständen, deren Territorialbesitz und Vermögen sehr stark divergierten, ermöglichen, in den Türkenkriegen und im
Kampf gegen die Hegemonie Ludwigs XIV. beachtliche militärische Leistungen zu erbringen,
womit zu Recht jenes karikierende, unzulässig verallgemeinerte Bild von der Reichsarmee als
»Reiß-Aus-Armee« korrigiert wird, das sich auf das unrühmliche Verhalten im Siebenjährigen
Krieg, vor allem auf die Schlacht bei Roßbach, gründet.
Das vom Schwäbischen Reichskreis aufgestellte Heer, dessen rechtliche Basis zum einen auf
Reichstagsbeschlüssen, zum anderen auf freiwilligen und somit ständisch-föderativen Vereinbarungen der Territorialherren beruhte, unterschied sich nach Storm hauptsächlich dadurch
von den Streitkräften anderer Mächte, daß es aus einer großen Anzahl von Kontingenten zusammengesetzt war, deren Kopfzahl obendrein - so z . B . im Jahr 1701 - zwischen rund 2000
und 4 Soldaten schwankte. Zudem war diese Streitmacht, die im Konfliktfall ein paar Tausend
Reiter und Infanteristen zählte, nur im Krieg wirklich präsent, weil die Einheiten und Verbände
im Frieden drastisch reduziert wurden und die Kontingente bei den Fürsten und Ständen im
Quartier lagen. Gemeinsame Übungen oder gemeinsames Exerzieren gab es kaum, ein Teil des
kostspieligen Kriegsgeräts wurde außerdem abgeschafft, kurz, mehr als größere Kader und gewisse Ausrüstungsvorräte wurden im Frieden nicht beibehalten. Als selbstverständlich muß
überdies gelten, daß diese Streitmacht nicht rasch kampfbereit sein konnte, hatte doch bei
Kriegsausbruch zunächst ein allgemeiner oder engerer Kreistag über die erforderlichen Mobilisierungsmaßnahmen zu beraten und zu beschließen, bevor an die Ausführung gegangen werden
konnte. Ein solches Heer entspricht gewiß nur bedingt unseren Vorstellungen vom miles perpetuus, indes gilt zu berücksichtigen, daß es im allgemeinen nicht die Aufgabe der Kreistruppen
war, auf sich alleingestellt im Feld eine Entscheidung zu suchen. Vielmehr kämpften sie meistens in Anlehnung an die Streitkräfte einer großen Macht, in der Regel zusammen mit Regimentern der casa d'Austria, die Rückhalt boten und deren Befehlshaber auch der maßgebende
Einfluß auf die Operationen zufiel.
Infolge ihrer komplizierten Struktur stößt man bei den Kreistruppen auf eine Fülle von Besonderheiten, die andere Armeen nicht kennen. Das fängt bei der Aufstellung der Kompanien an,
zu denen nicht selten ein halbes Dutzend oder mehr Stände mit Mannschaften konkurrierten,
setzt sich bei der verwickelten Vergabe der Prima-plana-Stellen fort, wirkt sich auf die schwer
zu vereinheitlichende Besoldung, Ausrüstung und Bewaffnung aus und macht sich ebenso beim
Einsatz bemerkbar, falls etwa die Truppen unerwartet für den territorialen Schutz beansprucht
wurden oder ein Landesherr plötzlich auf seine Hoheitsrechte pochte, um unbequemen Anforderungen zu entgehen oder eine zu weitreichende Integration zu blockieren. Konfessionelle Divergenzen, die zeitweise zur Formation »evangelischer« und »katholischer« Regimenter führten
oder bei der Besetzung höherer Offizierstellen ein paritätisches Vorgehen erzwangen, bezeugen
die zusätzlichen Schwierigkeiten, die in einem circulus mixtus erwuchsen. Vor allem aber bedeutete die Finanzierung des von den Ständen gemeinsam aufzubringenden, teuren Heeresbedarfs, der über die Rekrutierung von Kürassieren, Dragonern, Musketieren oder Grenadieren
hinausging, stets ein großes Problem, weil bei vielen Territorialherren neben den Mitteln oft
auch die Einsicht fehlte, der Streitmacht rechtzeitig angemessenes Gerät und qualifiziertes Personal zur Verfügung zu stellen. Das Proviant-, Fuhr- und Schiffsbrückenwesen, die Kommissariate und die Artillerie, die in dieser Zeit allmählich den zunftartigen Charakter verlor, hatten
darunter besonders zu leiden. Wenn sich die Regimenter des Schwäbischen und Fränkischen
Kreises unter dem Kommando des kaiserlichen Generalleutnants und Reichsgeneralfeldmarschalls Ludwig Wilhelm von Baden, des Türkenlouis, dennoch bewährten und beispielsweise
jahrelang die Oberrheinlinien gegen Frankreich verteidigten, so belegt dies, daß die Truppen
trotz ungünstiger Voraussetzungen durchaus in der Lage waren, sich mit den Verbänden der
Armierten zu messen.
In eindrucksvoller Weise hat Storm - gestützt auf die einschlägigen Quellen - die vielschichtige
Wehrverfassung des Schwäbischen Kreises in ihren rechtlichen Voraussetzungen und in der
konkreten Ausgestaltung herausgearbeitet und zugleich dargelegt, mit welchen Maßstäben eine
solche Armee zu messen ist, um sie gerecht beurteilen zu können. Für diese fundierte Studie, in
der die Führungsorgane des Kreises ebenso behandelt werden wie beispielsweise die Herkunft
der Offiziere oder das Dienstrecht der Soldaten, gebührt ihm Anerkennung. Gleichwohl soll
auf ein paar kritische Anmerkungen nicht verzichtet werden, die allerdings das Verdienst des
Verfassers nicht schmälern sollen.
Vorausgeschickt sei, daß die Untersuchung bei der Auswertung der Literatur nicht auf dem
neuesten Stand ist, weil die zwischen dem Abschluß des Werks und der Drucklegung erschienenen Publikationen nicht eingearbeitet wurden. Sodann ist auf einige sprachliche Eigentümlichkeiten hinzuweisen, die MißVerständnisse hervorrufen können. Dazu gehört der Titel, mit dem
Storm auf die zeitgenössische Terminologie zurückgegriffen hat und den unbefangenen Leser
zunächst insofern irreführt, als er ihm erst nach mehr als 100 Seiten Lektüre erläutert, daß hier
»Feldherr« nicht im heute gebräuchlichen Sinn, sondern im Sinn von Kriegs- und Dienstherr
gemeint ist. Zudem gefällt sich der Verfasser gelegentlich in veralteten oder ungewöhnlichen
Wortschöpfungen und Begriffen wie »wehrlich«, »abmehren«, »Standesführerwahl«, »Kreisführerwahl« oder schreibt von einer »evangelischen Kreiskasse« oder »katholischen Feldkästen«. Manchmal wendet Storm ferner in unzulässiger Weise heutige Begriffe auf die Verhältnisse im 17. und 18. Jahrhundert an. So muß es zumindest als fragwürdig erscheinen, von
»Sachnormen« bei dem wenig erfolgreichen Bestreben zur Vereinheitlichung der Waffen und
der Ausrüstung zu reden (S. 4 9 2 , 4 9 6 ). O b es sinnvoll ist, die Zahl der »Rohre« ( = Geschütze) in
Beziehung zur Truppenstärke zu setzen und daraus eine Art Schlüsselwert zu errechnen (S. 437
ff.), ohne zu prüfen, ob und wie diese Stücke zum Einsatz kamen, muß angezweifelt werden,
zumal für die Kreisstände überwiegend finanzielle Erwägungen, nicht aber der Wunsch nach einem ausgewogenen Verhältnis zwischen der Feuerkraft der Infanterie und der Artillerie eine
Rolle spielte. Anfechtbar sind darüber hinaus die Tabellen über den Territorialbesitz und die
Untertanen der schwäbischen Fürsten und Stände (S. 51 ff.), weil deren teils frappierend genauen Werte eine Exaktheit vorgaukeln, die es nicht gibt und nicht gegeben haben kann, denn
im Gebiet der territoria inclausa waren die Hoheitsansprüche nur allzuoft strittig. Zwar weiß
Storm darum, doch nennt er trotzdem Zahlen bis zur Dezimalstelle hinter dem Komma oder
weist bei zwei, drei Ständen eine auf den Kopf genaue Untertanenschaft nach. Außerdem war er
allzusehr bedacht, die Wehrverfassung des Kreises in eine systematische Ordnung zu bringen,
wobei anscheinend der Wirklichkeit hin und wieder Gewalt angetan wird. D e facto jedenfalls
dürften viele Stände zwischen den Funktionen des vormaligen Kreisobristen und des späteren
Kreisgeneralfeldmarschalls keinen grundlegenden Unterschied entdeckt haben. Problematisch
erscheint es gleichfalls, eine Kategorie »Sondertruppen« zu konstruieren und zu diesen die Artillerie, das Proviantfuhrwesen, das Backwesen und das Gesundheitswesen zu rechnen, die
Kriegs- und Proviantkommissare und das Kriegszahlamt indes auszunehmen. Bernhard
Sicken
Anmerkung
1
F. Härtung: Deutsche Verfassungsgeschichte vom 15. Jahrhundert bis zur Gegenwart. Stuttgart e 1 9 6 4 ,
S. 42.
Antonia Fraser: Cromwell. The Lord Protector. New York: Knopf 1973. XX, 774 Seiten
Cromwell: A profile. Edited by Ivan Roots. London, Basingstoke: Macmillan 1973.
XVIII, 237 Seiten (= World Profiles.)
Schon zu Lebzeiten umstritten und Gegenstand leidenschaftlicher Schmähschriften oder aber
emphatischer Loblieder, hat die Gestalt Oliver Cromwells auch heute noch erstaunliche Anziehungskraft auf Historiker. Verfolgt man die lange Kette der Cromwell-Literatur seit der Restaurationszeit, so fällt ins Auge, daß kaum einer seiner Biographen Cromwell wirklich distanziert gegenüberstand.
Antonia Fraser, bekannt geworden durch ihre populärwissenschaftliche »Mary Queen of Scots«
(London e 1969), hat es sich zum Ziel gesetzt, in ihrer breitangelegten Darstellung den Menschen
Cromwell aus der Obskurität zu holen. Ihr Versuch, Cromwell zu »vermenschlichen« (S.
XIII), soll eine Art Gegengewicht zu den zahlreichen, in der heutigen Forschung dominierenden Arbeiten bilden, die sich auf die politischen, sozialen und wirtschaftlichen Aspekte der Zeit
konzentrieren. Leider macht die Verfasserin es dem Leser schwer, dieser Richtlinie zu folgen. In
ihrem Bestreben, die Person ihres Helden in die Geschichte der englischen Bürgerkriege und des
Commonwealth einzubetten, hat die große Menge des verarbeiteten Materials sie offenbar dazu
verleitet, ihre Darstellung übermäßig ausufern zu lassen.
Auf der einen Seite macht sie aus ihrer fast grenzenlosen Bewunderung Cromwells kein Hehl;
der Satz, Cromwell sei mit Sicherheit der »größte Engländer«, fließt ihr leicht aus der Feder.
Andererseits bemüht sie sich durchaus um kritische Distanz. Dies wird in der unermüdlichen
Auseinandersetzung mit Mythen, Lügen, Diffamierungen, Lobpreisungen und sonstigen
Zeugnissen, die ihren Helden betreffen, deutlich.
Dieses Ringen mit dem Detail, die häufigen Abschweifungen und überlange Schlachtenbeschreibungen machen das Buch zum Teil schwer lesbar und werden dem Anspruch, ein Bild
»des Menschen« Oliver Cromwell zu zeichnen, nicht gerecht. Auch die Spannung zwischen
Identifizierung und kritischer Analyse wird eigentlich nicht fruchtbar. Als Beispiel sei hier nur
die Darstellung der Westindien-Expedition angeführt. Die kritischen Ergebnisse der neuesten
Forschung sind zwar berücksichtigt worden, werden aber letztlich dann wieder durch Hinweise
in Frage gestellt, wie z. B., die Außenpolitik auch der Stuarts sei kostspielig gewesen, oder auch
mit der Bemerkung, selbst Edward Hyde, ein gewiß unverdächtiger Zeuge, habe festgestellt,
»that Cromwell's greatness at home was but a shadow of the glory he had abroad« (S. 553).
Auch für Ivan Roots steht die »Ungreifbarkeit« der Person Cromwells, in der ein ganzes Zeitalter sich widerspiegele, im Vordergrund, wie er in der Einleitung betont. In erfreulicher Einschätzung der Möglichkeiten der historischen Biographie betont er die Zeitgebundenheit des
Urteils. Dennoch glaubt er ein wesentliches Merkmal auszumachen, wenn er das »Genie«
Cromwells mit Carlyle als »a transcendent capacity of taking trouble« (S. XI) definiert. Cromwell stellt sich nicht als ein »Macher« dar, sondern als ein Wartender, der in sich »hineinhorcht«, mit der Realität des Augenblickes lebt und diese Realität auszuschöpfen sucht.
So ist es nur konsequent, wenn sich die meisten Arbeiten des Bandes besonders mit dieser Wirklichkeit der Cromwell-Zeit befassen, mit Ausnahme der Studie H . W. Kochs über »Cromwell's
Genius« und Ernst Barkers: The Achievement of Oliver Cromwell, die beide ihre wesentlichen
Impulse aus der Leistung des »Soldaten« Cromwell beziehen.
Paul H . Hardacre behandelt das Verhältnis der Royalisten zu Cromwell; Austin Woolrich beschreibt Cromwells »harshest lesson« in seiner Studie über die Herrschaft der »Heiligen«, und
Hugh Trevor-Roper ist mit seinem bekannten Aufsatz »Oliver Cromwell and his parliaments«
vertreten. Vor allem die beiden letztgenannten Arbeiten machen deutlich, daß Cromwells fehlende Bereitschaft oder Fähigkeit, seinen Anhang parlamentarisch zu organisieren oder, allgemeiner ausgedrückt, sich den Spielregeln politischer Planung unterzuordnen, mitverantwortlich für sein »Scheitern« war. Trevor-Ropers Argument, Cromwells Kardinalfehler sei sein Unverständnis der »essentials« elisabethanischer Parlamentspolitik gewesen, klingt zwar originell,
geht aber doch etwas an der Realität vorbei, denn der Pfeiler seiner Macht war das Heer.
Dies ändert nichts an der Richtigkeit der Beobachtung, daß die Vorstellung Cromwells, ein Parlament der »back benchers« sei auf die Dauer möglich, illusionär war. Den konservativen Habitus der Commonwealth-Ära zeigt G. D. Ramsay in seiner Arbeit »Industrial Laisser-Faire and
the policy of Cromwell«, wenn er die auf durchaus traditioneller ( = »royalistischer«) Linie liegende Politik des Lord Protektors gegenüber den »Chartered Cities« und Gilden verfolgt und so
seine Vermutung, die Wurzeln gewollter Laisser-Faire-Politik lägen in der Revolutionszeit,
nicht bestätigt sieht.
Im Gegensatz zu zahlreichen Untersuchungen der Cromwellschen Außenpolitik ist »The Idea
of a protestant foreign Policy« von Roger Crabtree bemerkenswert differenziert. Crabtree
zeigt, daß sich Cromwell weder als religiöser Fanatiker und demzufolge außenpolitischer Ignorant und Phantast begreifen läßt, sondern daß herrschaftssichernde Momente (Stuart-Gefahr),
außenpolitische und religiöse Komponenten zusammenspielten.
Die in Cromwells Denken genuine Mischung kühler Abwägung und religiös begründeter. Suche
nach »Zeichen« kann jedenfalls als Faktum genommen werden und entzieht sich ab einem gewissen Punkt der rationalen Analyse.
Ähnlich argumentiert Christopher Hill in einem Auszug seiner jüngsten Cromwell-Biographie 1 , wenn er darlegt, daß der Glaube an die Vorsehung nicht als ein unmittelbarer Widerspruch zur Idee menschlicher Freiheit zu sehen ist.
Auch kommende Biographen des Lord Protektors, meint D . H . Pennington in seinem abschließenden Überblick der Cromwell-Historiographie, werden letztlich finden, »wonach sie suchen«. Die Palette des vorliegenden Bandes, der durch einen kurzen bibliographischen Anhang
abgerundet wird, sollte dazu beitragen, einseitige Urteile zu vermeiden. Die dem 18. Jahrhundert entstammende Vermutung Dr. Johnsons: »all that can be told of him is already in print« als
Resümee der Cromwell-Forschung ist schon längst ad absurdum geführt worden. Dem Herausgeber mag man jedenfalls gratulieren, daß seine Bemerkung, Cromwell gehöre zu den bekanntesten, doch am wenigsten »begriffenen« Personen der englischen Geschichte, durch den
vorliegenden Band zumindest eine Relativierung erfahren hat.
Hans-Ch.
Junge
Anmerkung
1
CH. H i l l : G o d ' s Englishman. Oliver Cromwell and the English Revolution. L o n d o n 1970.
Frederick the Great. A profile. Edited by Peter Paret. London: Macmillan 1972. X X I ,
249 Seiten ( = World Profiles.)
Obwohl die gegenwärtige Historiographie ihrer Intention nach eher antibiographisch orientiert
ist und dazu neigt, statt der »Persönlichkeiten« die geschichtlichen »Strukturen« zu betonen,
hat sie doch gerade in den letzten Jahren eine ausgedehnte Friedrich-Literatur von allerdings
sehr unterschiedlicher Qualität hervorgebracht. Mit einiger Berechtigung darf man von einer
Art Friedrich-Renaissance sprechen, die inzwischen über den deutschen Sprachraum hinausgegriffen hat und nach Westeuropa auch die Vereinigten Staaten zu erfassen scheint. In Deutschland hat vor allem das anklägerische Friedrich-Buch von Augstein (1968) breitere Aufmerksamkeit gefunden; wissenschaftlichen Ertrag brachte hingegen die Monographie von Walther Hubatsch über Friedrich und die preußische Verwaltung. Aus Frankreich liegt uns seit 1973 die
glänzend geschriebene Biographie des Königs von Pierre Gaxotte in einer einfühlsamen deutschen Ubersetzung vor. Aus Großbritannien haben wir neben der bekannten Studie von G. P.
Gooch (1947) gleich zwei neue Versuche einer Lebensbeschreibung, die stark psychologisierende Deutung von Edith Simon und die 1973 überflüssigerweise übersetzte, oberflächliche
Darstellung von Nancy Mitford.
Diesen biographisch orientierten Werken gesellt sich die von dem in Stanford lehrenden Historiker Peter Paret besorgte vorliegende Friedrich-Anthologie zu. Auch sie verfolgt die Absicht,
durch einen Wald von Legenden und späteren Interpretationen hindurch den preußischen König als Menschen und Staatsmann des aufgeklärten 18. Jahrhunderts zu vergegenwärtigen und
aus dem historischen Bezugsrahmen heraus zu deuten bzw. zu verstehen.
Um dieses Ziel zu erreichen, wählt der durch seine Studie zur Militärreform mit der preußischen
Geschichte des ausgehenden 18. Jahrhunderts vertraute Herausgeber aus der bisherigen wissenschaftlichen Friedrich-Literatur diejenigen Stücke aus, die ihm für eine bestimmte Entwick-
lungsphase des Königs bzw. für einen bestimmten Sachzusammenhang besonders aussagekräftig erscheinen. Er ergänzt sie um zwei zeitgenössische Berichte über eine Inspektionsreise des
Königs 1779 und über die Schlacht bei Hochkirch 1758, zwei gegensätzliche, aber betont kritische Deutungsversuche, sowie um einen ganz knappen Lebenslaufund eine kurze, im einzelnen
freilich nicht ganz sorgfältig gearbeitete Bibliographie raisonée.
Von den vier Hauptteilen der Sammlung soll der 1. die Biographie Friedrichs im engeren Sinne
beleuchten: Er enthält zunächst die unübertroffene Deutung, die Carl Hinrichs in seiner Einleitung zum »Kronprinzenprozeß« dem für die Persönlichkeitsentfaltung zentralen Konflikt zwischen dem königlichen Vater und seinem so anders gearteten Sohn gegeben hat, ferner ein bezeichnendes Stimmungsbild des aus dem Siebenjährigen Krieg nach Berlin zurückkehrenden
Königs aus der Heinrich-Biographie von Chester V. Easum und schließlich eine Schilderung der
Altersaktivitäten und -interessen des Monarchen aus den Friedrich-Studien von George P.
Gooch.
Der 2. Hauptteil soll dem Leser Friedrich als Regenten, Feldherrn und Staatsmann nahebringen.
Dazu bedient sich Paret einer Auswahl aus der aufschlußreichen Untersuchung von Walter L.
Dorn über die preußische Verwaltung, ferner des der Kriegführung des Königs gewidmeten Abschnitts in Gerhard Ritters Friedrich-Buch und schließlich Friedrich Meineckes tiefdringender
Analyse des Verhältnisses von Philosoph und Staatsmann in »Die Idee der Staatsräson«.
Der 3. Hauptteil »Friedrich und die Aufklärung« beschränkt sich auf seine Stellung zur preußischen Akademie, wie sie sich bei Wilhelm Dilthey: Studien zur Geschichte des deutschen Geistes. 21942 (= Dilthey: Gesammelte Schriften. Bd 3.) findet, und auf sein distanziertes Verhältnis zur deutschen Literatur, das Friedrich Gundolf adäquat interpretiert hat.
Der 4. Hauptteil schließlich dokumentiert die tatsächliche Variationsbreite bisheriger Versuche,
die Stellung Friedrichs im Ablauf der preußisch-deutschen Geschichte zu bestimmen, nur sehr
begrenzt, indem er Franz Mehrings polemischen, nichtsdestoweniger brillanten Artikel »Ein
aufgeklärter Despot?« (Die Neue Zeit. Jg 1912) der großdeutsch-habsburgisch orientierten
Friedrich-Kritik Heinrich Ritter v. Srbiks aus dessen »Deutscher Einheit« gegenüberstellt.
Zwei exponierte, aber historiographisch selbst schon überlebte Positionen, die freilich den Hintergrund auch noch für die marxistische bzw. reichsbezogen argumentierende Friedrich-Kritik
unserer Tage bilden, sind also das Auskunftsmittel, auf das der Leser zu einer abschließenden
Würdigung von Friedrichs Stellung in der deutschen Geschichte verwiesen wird. Dennoch will
es scheinen, als gehörten die Sympathien des Herausgebers eher der ebenso verständnisvollen
wie subtilen Deutung Friedrich Meineckes, die in einem Dualismus zwischen »Idee« und
»Macht«, zwischen Staatsmann und Philosoph endet und deshalb nicht zu einer einheitlichen
Persönlichkeitsdeutung des Preußenkönigs gelangt, wie sie wohl versucht werden müßte.
Bedenkt man die Schwierigkeiten, die sich einem derartigen Unterfangen in den Weg stellen Paret selbst formuliert sie in dem Satz (S. XIII): »We will find it extremely difficult to blend the
complexities of his character and behavior into an assessment that is balanced as well as comprehensive« - , und zieht man vergleichsweise die oben skizzierte neuere biographische Literatur
heran, so wird man dem Herausgeber bestätigen dürfen, daß es ihm gelungen ist, eine wohlproportionierte Anthologie vorzulegen. Sie spiegelt den Stand der modernen Spezialforschung
adäquat wider und sucht der komplizierten Gestalt des Preußenkönigs im Rahmen seiner Zeit
gerecht zu werden. Der Rezensent ist daher sehr geneigt, dieser überlegten Auswahledition den
Vorrang vor zahlreichen Gesamtdarstellungen zu geben, deren äußere Einheitlichkeit durch
Oberflächlichkeit oder mangelndes Problembewußtsein erkauft wird. Er empfiehlt sie angelegentlich als Einführung in das »Problem« Friedrich.
Peter Baumgart
Hartmut Rudolph: Das evangelische Militärkirchenwesen in Preußen. Die Entwicklung
seinerVerfassung und Organisation vom Absolutismus bis zum Vorabend des I. Weltkrieges. Mit einem dokumentarischen Anhang. Göttingen: Vandenhoeck & Ruprecht
1973. 433 Seiten (= Studien zur Theologie und Geistesgeschichte des 19. Jahrhunderts.
Bd 8.)
Das evangelische Militärkirchenwesen in Preußen ist - sieht man von einigen überwiegend älte-
ren kirchenrechtlichen Abhandlungen ab - bisher weder von der militar- noch der kirchengeschichtlichen Forschung in zureichender Weise behandelt worden. Diese Lücke wird für das
preußische Militärkirchenwesen des 19. Jahrhunderts durch die vorliegende Heidelberger Dissertation weitgehend ausgefüllt, wenn auch bei dem Mangel an Voruntersuchungen und der nur
partiellen Erschließung der Quellen eine umfassende Darstellung noch unerreicht blieb. Eine
solche würde auch bei einer das Militärkirchenwesen isolierenden, längsschnittartig die Kontinuität verfolgenden Untersuchung verfehlt werden, worauf der Verfasser mit Recht hinweist,
sondern wäre nur in einer sehr weitgespannten Analyse sowohl der militar- wie der kirchen- und
theologiegeschichtlichen Entwicklung sinnvoll zu thematisieren.
Der Verfasser setzt mit einer kurzen Skizzierung des Militärkirchenwesens im absolutistischen
Preußen ein und bestätigt dabei nachdrücklich die Richtigkeit des Urteils von Otto Hintze, wonach das Kirchenregiment zweifellos als eine der schwächsten Seiten des preußischen Militärstaats angesehen werden muß. Dies gilt in besonderem Maße für die damals in nahezu vollständigem Umfang staatskirchlich verfaßte und gegenüber der Landeskirche verselbständigte Militärkirche, auf die neben dem obersten Militärgeistlichen, dem Feldpropst, Offiziere und Militärjuristen, nicht aber landeskirchliche Instanzen entscheidenden Einfluß ausübten. Die eklatanten Mißstände waren in der Zeit des Rationalismus allerdings innerhalb der bis zum Ende des
18. Jahrhunderts als organisatorische Einheit fast vollständig paralysierten Landeskirche nicht
minder verbreitet als in der streng auf die Bedürfnisse der Armee ausgerichteten, in den Heeresorganismus integrierten Militärkirche. Auch dürfte die Annahme ungesichert erscheinen, daß
erst die Instrumentalisierung der Religion im Interesse des Zusammenhalts der Armee und des
ihr zugrundeliegenden Herrschaftskonsenses der Zuwendung der Landbevölkerung zur Kirche
ein Ende gemacht habe.
Breiten Raum nimmt die Darstellung der kirchlichen Neuordnung in der Reformzeit nach dem
militärischen Zusammenbruch Preußens ein. Es ist ein bemerkenswertes Indiz für den Mangel
kirchlicher Gestaltungskraft, daß den Reformvorstellungen im staatlich-politischen Bereich, im
Militärwesen und auf dem Bildungssektor kein adäquates Reformkonzept für die kirchlichen
Verhältnisse entgegengesetzt werden konnte, sieht man einmal von Schleiermacher ab. Das Rezept der staatlichen Reformer für die Kirche war die völlige Verstaatlichung! Der auf diese Weise
ihrer Selbständigkeit beraubten Kirche wurde die Militärseelsorge (Humboldt: eine »verderbliche Pflanzschule des geistlichen Standes«) weitgehend inkorporiert. Nicht zwingend scheint
der Rückschluß des Verfassers von der Usurpierung der Kirche für die nationale Erhebung während der Befreiungskriege als einer direkten Folge ihrer staatskirchlichen Verfassung.
Die Reorganisationsmaßnahmen der Reformzeit überdauerten kaum den Beginn der 1815 einsetzenden Restauration. Das Verhältnis von Staat und Kirche blieb vorerst ungeklärt, die Integration der Militärkirche in die zivile Landeskirche wurde schrittweise zurückgenommen. Uber
die allgemeine Kausalverknüpfung dieser Maßnahme mit der Funktion der Religion für die
ideologische Stützung und Ausbreitung restaurativer Vorstellungen hinaus hebt der Verfasser
den Stellenwert der Militärgemeinden als Vehikel zur Durchsetzung staatlicher Interessen im
gesamtkirchlichen Bereich hervor, in exemplarischer Weise bei der Schaffung der preußischen
Union.
Den Schwerpunkt der Darstellung bildet das Militärkirchenwesen zwischen der Revolution von
1848 und dem Ersten Weltkrieg, das der Verfasser vor dem knapp angedeuteten Hintergrund
der Veränderungen in der Armee und der evangelischen Landeskirche untersucht. Dieser Abschnitt wird weitgehend bestimmt durch perennierende Auseinandersetzungen zwischen dem
1850 begründeten Evangelischen Oberkirchenrat und den beteiligten staatlichen Instanzen um
den Einfluß auf die Militärgemeinden.
Die Bemühungen des Oberkirchenrats blieben aus mehreren Gründen erfolglos: einmal wegen
der seit 1815 und verstärkt wieder seit dem Abbau des Kulturkampfes erforderlichen Rücksichtnahme auf den Grundsatz staatlicher Parität (hatte es doch bis zur Mitte des 19. Jahrhunderts keine festangestellten katholischen Militärgeistlichen gegeben, obwohl die katholischen
Soldaten den Militärgemeinden angehörten!); zweitens wegen der Beschränkung der Kompetenzen des Oberkirchenrats auf die altpreußischen Provinzen und drittens wegen der mit zunehmender »Militarisierung« immer stärker hervortretenden Abneigung, eigenständige kirchli-
che Einflußnahme, sofern sie nicht staatlich gesteuert und kontrolliert wurde, zuzulassen. Das
Ergebnis war jedenfalls, daß mit dem Erlaß der »Evgl. militärkirchlichen Dienstordnung« von
1902 die wesentlichen kirchenregimentlichen Bindungen der Militärgemeinden zur Landeskirche gelöst wurden zugunsten der »Militarisierung des Militärkirchenregiments«.
Es mag verwundern, daß in dieser sehr gründlichen Studie eines Theologen Reorganisationsmaßnahmen und Kompetenzstreit derart zentral im Vordergrund stehen. Der Verfasser vermag
jedoch sein Vorgehen plausibel zu machen. Denn die Kraft und Ausstrahlung der Kirche wurde
in ganz unverhältnismäßig hohem Maße von solchen Emanzipationsbestrebungen von staatlichen Einflüssen absorbiert. Der Verfasser hätte dennoch seine durchaus vorhandenen Ansätze
zur Charakterisierung der unmittelbaren seelsorgerlichen Tätigkeit einzelner Feldprediger weiter ausführen sollen, vor allem im letzten Abschnitt. Es wäre von Interesse zu erfahren, ob sich
die Militärgeistlichen in der zweiten Hälfte des 19. Jahrhunderts dem Offiziersmilieu mit einem
sehr veräußerlichten Religionsverständnis, nicht selten sehr platt zur Stabilisierung bestehender
Klassenstrukturen instrumentalisiert, widerspruchslos angepaßt haben. Daß man unter den Militärgeistlichen nicht in ähnlichem Umfang eine gegen Kapital und Grundbesitz gerichtete, auch
vor Staats- und Kircheninstanzen nicht Halt machende Protestbewegung wie unter den Geistlichen der Landeskirche in den 90er Jahren annehmen kann, läßt sich schon auf Grund ihrer privilegierten Stellung annehmen. Die Anstellung als Militärgeistlicher begünstigte den Aufstieg in
die Leitung des Kirchenregiments, begründete zumindest eine Anwartschaft auf eine der höher
dotierten Pfarrstellen der Landeskirche.
Insofern wurden Amtsverständnis, Mentalität und Gesellschaftsanschauung der Militärgeistlichen auch für die Landeskirche unmittelbar relevant, (man vermißt ζ. B. einen Hinweis auf den
ehemaligen Militärgeistlichen in Metz, Adolf Stoecker!), soweit ihr nicht ohnehin Royalismus
als religiöse Kardinaltugend galt und sie deshalb der Indoktrination kaum bedurfte. Die Hierarchisierung der Geistlichen findet ihren stärksten Ausdruck in dem lange umstrittenen Amt des
Feldpropstes, der am Ende des 19. Jahrhunderts einen straffen Einfluß auf die Militärgeistlichen
ausübt und gegenüber dem Kirchenregiment - sogar als Repräsentant eines spezifischen konfessionellen Standorts der Militärseelsorge - eine merkliche Distanz wahrt.
Ein breiter dokumentarischer Anhang mit instruktivem, schwer zugänglichem Quellenmaterial
ergänzt die Darstellung.
Pollmann
Soldaten im bunten Rock. (1-3 : Prestige de l'uniforme. The Glamour of uniforme.) 1—4.
Stuttgart: Frankh 1968-72.
1. Hans-Joachim
Ullrich: Die preußische Armee unter Friedrich Wilhelm II. und Friedrich Wilhelm III., 1786-1807. L'Armée prussienne sous Frédéric-Guillaume II et Frédéric-Guillaume III, 1786-1807. The Prussian Army under Frederick William II and
Frederick William III, 1786-1807. Französische Übertragung: Paul Martin. Englische
Übertragung: René North. 1968. 34 Seiten
2. Paul Martin, Hans-Joachim
Ullrich: Die französische Armee 1789-1807. L'Armée
française 1789-1807. The French Army 1789-1807. Englische Übertragung René North.
1969. 40 Seiten
3. Hans-Joachim
Ullrich: Die preußische Armee 1840-1871. L'Armée prussienne
1840-1871. The Prussian Army 1840-1871. Französische Übertragung: Paul Martin.
Englische Übertragung: René North. 1970. 38 Seiten
4. Hans-Joachim
Ullrich: Die preußische Armee 1808-1839. 1972. 15 Seiten
Andrew Mollo, Malcolm McGregor: Armeeuniformen des 2. Weltkriegs in Farbe. Deutsche Übersetzung von Bernd und Karin Nosky. München: Heyne 1974. 196 Seiten
137
Jede dieser vier Mappen enthält zu der Einzelblattsammlung von Wiedergaben durchweg zeitgenössischer farbiger Bilder und Stiche einen Text, der den betreffenden militärgeschichtlichen
und uniformkundlichen Zusammenhang kurz umreißt, die Bilder erklärt und mit Schwarzweißzeichnungen auf besondere Uniformdetails hinweist. Die Bilder sind nicht nur nach ihrem
uniformgeschichtlichen Informationswert, sondern auch im Hinblick auf ihre Verwendung als
Wandschmuck nach ihrer künstlerischen Qualität ausgesucht worden. Die Veröffentlichungen
beschränken sich nicht auf die Wiedergabe der reglementsmäßigen Bekleidung, sondern geben
auch Einblick in das historische Soldatenleben in Krieg und Frieden, beim Dienst und in der
Freizeit. Lebendig und wirklichkeitsnah, gelegentlich sogar humorvoll bis zur Karikatur sind
vor allem die französischen, individualistisch akzentuierten Bilder. Bei den preußischen wird
dagegen der Hauptwert auf die korrekte Vorstellung der Uniform in ihren Details gelegt. Sie
sind also auch dort, wo sie einen genrehaften Bezug aufweisen, mehr Uniformbilder im engeren
Sinne.
Die begleitenden Texte behandeln das Thema lediglich unter dem Aspekt der betreffenden
Armeen und gehen auf die Gesamtentwicklung nicht ein. Welche Bedeutung der Uniformentwicklung bei den einzelnen Armeen für die Gesamtentwicklung zukommt, wäre freilich nur
durch eine entsprechende Einordnung zu zeigen. Die drei, von dem verstorbenen Hans-Joachim Ullrich für Preußen bearbeiteten Phasen der Uniformentwicklung ließen sich, mit gewissen zeitlichen Verschiebungen, für ganz Europa nachweisen. Kennzeichnend für die Verhältnisse in Preußen war es, daß während der Scharnhorstschen Heeresreform hier eine völlig neue,
am russischen Vorbild orientierte Militärbekleidung eingeführt wurde. In Frankreich, dessen
Uniformstil dann a\ich für viele andere Armeen - auch außereuropäische - maßgebend wurde,
verlief die Entwicklung von der barock geprägten Montur des 18. Jahrhunderts bis zu der der ersten Hälfte des 19. Jahrhunderts trotz der Revolution kontinuierlicher, mehr in fließenden
Übergängen. Unter Napoleon I. erreichte die militärische Prachtentfaltung ihren Höhepunkt.
Im Vergleich damit wirkten die preußischen Uniformen, die nicht nur verhältnismäßig praktisch, sondern auch billig waren, sehr schlicht.
Während der Restauration blieben die Uniformen in den Grundzügen der napoleonischen
Kriege erhalten. Es kam zu manchen Angleichungen, manchen Vereinfachungen, aber auch in
manchen Ländern zu noch reicheren Ausstattungen, jedenfalls der Paradeuniformen. Uberall
strebten die Armeen eine schlanke, straffe, spezifisch militärische Eleganz an, mit der sich der
Soldat betont von der bürgerlichen Behäbigkeit distanzierte. Damit ist aber auch gesagt, daß für
das Äußere des Soldaten das Parade-Ideal und nicht die feldmäßigen Gegebenheiten maßgebend
wurden. Diesem Ideal entsprach dieses Äußere mit den sehr hohen Tschakos, Helmen und Kragen, den knappen Frackjacken und den langen engen Hosen auf eine zweifellos männlichere Art
nicht weniger als die puppenhafte Aufmachung des Rokoko-Soldaten.
Als sich dann um die Mitte des 19. Jahrhunderts überall ein neuer Uniformstil durchsetzte, stellten die neuen Bekleidungen und Ausrüstungen in vieler Hinsicht das genaue Gegenteil der bisherigen Art dar. Mit den plumpen, unten grotesk abstehenden neuen Uniformröcken machten
die Soldaten keinen sehr ansprechenden Eindruck. In Preußen wurden sie in Erinnerung an das
entsprechende Kleidungsstück der alten Ritter »Waffenröcke« g e n a n n t - w i e man unter König
Friedrich Wilhelm IV. ja überhaupt rationelle Zweckmäßigkeit und romantische Emotionen zu
verbinden suchte und auch Fabrikscbernsteine mit Burgzinnen drapierte. Immerhin waren die
neuen Uniformmodelle bequemer als die alten, nachdem diese so beklemmend eng geworden
waren, und schützten besser vor der Witterung. Das erste Modell der dann viel nachgeahmten
preußischen »Pickelhaube« war, ästhetisch gesehen, ein Monstrum, aber es schützte besser gegen Säbelhiebe und Kolbenschläge als die bisherigen Tschako- und Helmformen. Man war mehr
als vorher auf das Wohlbefinden des Soldaten bedacht, was sich auch an einer verbesserten Trageweise des Marschgepäcks zeigte. Von der Basis dieses Uniformmodells aus vollzog sich dann
in Preußen und später in ganz Deutschland die weitere Entwicklung nicht nur bis zum Jahre
1871, sondern in Ausläufern bis in den Ersten Weltkrieg hinein. Auch die meisten übrigen europäischen Staaten entwickelten ihre Uniformen zunächst auf der Grundlage der Typen aus den
1840er bis 1860er Jahren weiter. Hierbei wurden allgemein die Waffenröcke kürzer, die Kopfbedeckungen leichter und niedriger, das Aussehen des Soldaten dadurch gefälliger.
So sehr man auch um einen Ausgleich von repräsentativer Wirkung und am Felddienst orientierter Zweckmäßigkeit bemüht war, auf die Dauer ließen sich die damit verbundenen Kompromisse nicht durchhalten. Während die ersten Khaki-Uniformen bereits Mitte des 19. Jahrhunderts bei britischen Kolonialtruppen in Indien auftauchten, zog das deutsche Heer in den Ersten
Weltkrieg zwar nahezu durchgehend in Feldgrau, aber mit Feldanzügen im Schnitt der farbigen
Friedensuniformen. Die französische Armee erschien 1914 sogar noch bunt auf dem Schlacht-
feld. Hierbei ist jedoch zu bedenken, daß vieles, was manchem heute an den alten bunten Uniformen als Kriterium bloßen militärischen »Imponiergehabes« erscheint, in taktischen und anderen, zumindest ursprünglich funktionalen Gründen seine Erklärung findet. So waren vor allem die leuchtenden Farben und Abzeichen als Erkennungssymbole für den auf verhältnismäßig
kleinem Raum geschlossen geführten Kampf notwendig gewesen. Die tiefgreifenden waffentechnischen und taktischen Veränderungen seit der Mitte des 19. Jahrhunderts, die Auflösung
der geschlossenen Kampfkörper, die damit verbundene viel stärkere Ausnutzung des Geländes
bedingten schließlich eine tarnfarbene Feldbekleidung, in der sich der Soldat möglichst ungehindert und ungesehen bewegen konnte. Diese neuen Feldanzüge waren aber für die friedliche
militärische Repräsentation denkbar ungeeignet. So erhielt jeder Soldat zusätzlich zu seinem
Feldanzug noch eine weitere, mehr und mehr in Farbe und Schnitt von ihr abweichende Ausgeh- und Paradeuniform.
Mollo und McGregor befassen sich ausschließlich mit Felduniformen. Sie liefern keine Illustrationen der betreffenden Bekleidungsvorschriften, sondern legen eine Bilddokumentaion darüber vor, wie die Soldaten im Zweiten Weltkrieg tatsächlich aussahen, also wie Bekleidung und
Ausrüstung tatsächlich getragen wurden. Die farbigen, in der Qualität hervorragenden BilderEinzelfiguren auf Tafeln - sind nach Fotografien hergestellt worden. Wo es möglich war, wurden auch die betreffenden Truppenteile, ja sogar die Namen von Soldaten - und nicht nur die
von Generälen - genannt. In dieser unmittelbaren, durch kein falsches Pathos hochstilisierten,
durch keine Schönfärberei oder nachträgliche »Korrektur« verfälschten Realistik liegt der besondere, nicht nur uniformkundliche Wert dieser Arbeit. Hier sind nicht nur Kleidung und
Ausrüstung, sondern auch Haltung und Gesichtsausdruck aus den verschiedensten Situationen
des Krieges heraus eingefangen und zu einer differenzierten Typik verdichtet worden, mit der
man weit über die bloße Uniformdarstellung hinausgelangte.
Nach Vorwort, Einleitung und einer Erklärung der Fachausdrücke werden die Feldanzüge der
meisten Länder beschrieben, die am Zweiten Weltkrieg teilgenommen haben. Daran angeschlossen ist der nach Feldzügen geordnete Bildteil. Er wird ergänzt durch Tafeln, auf denen die
im Kriege auf beiden Seiten benutzten Helme und Infanteriewaffen sowie die persönlichen Ausrüstungsstücke der Soldaten genau abgebildet sind. Am Schluß findet man dann noch die Erklärungen für die einzelnen Figuren.
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the FT with no preferential altitudes identified during two soundings (n°12 and 29), and three profiles do not show any indication of nucleation (n°9, 10 and 13). The vertical limits of the nucleation process are clearly visible for profiles n°6 and 28. For profile n°6, nucleation is observed between 1680 and 3170 m, with concentrations significantly increased between 2400 and 2900 m. During sounding n°28, nucleation is Fig. 6. Statistics on the detection of significant particle concentration in the size range 5 – 10 nm as a function of altitude (left panel, the number of data points is indicated on the plot for each altitude range). Corresponding median concentrations are reported on the right panel; left and right limits of the error bars stand for the 1 st and 3rd quartile, respectively. . In order to further investigate the vertical extension of nucleation, all measurements were considered, i.e. N5-10 concentrations measured during vertical soundings and also at constant altitude levels. Fig. 6 (left panel) shows the percentage of N5-10 concentrations exceeding the threshold value as a function of altitude range. As previously suggested by Fig. 5, nucleation seems to be favoured above 1000 m. In fact, below 1000 m, less than 4% of the recorded concentrations were found to be significant. In contrast, nucleation probability is increased by 10 above 1000 m, and it is worth noticing that almost 50% of the concentrations obtained between 2000 and 3000 m exceed the threshold value, indicating that these altitudes could more especially favour the occurrence of nucleation. However, it seems that when nucleation events are detected, the number of nucleated particles does not significantly vary with altitude, especially above 500 m, where median concentrations are in the range 309 – 376 cm-3 (Fig. 6, right panel). Below 500 m, N5-10 are slightly increased and show higher variability, which might be caused by more inhomogeneous conditions found in this part of the atmosphere compared to higher altitudes. The fact that nucleation is favoured in the FT compared to the BL contradicts the results by Crumeyrolle et al. (2010), who found that NPF events were limited to the top of the BL in the North Sea. However it is worth noticing that the number of vertical profiles included in the Crumeyrolles study was limited (13 profiles), most of them were performed close to the coast. iv. Why such a vertical extension? The purpose of this section is to investigate atmospheric parameters which are expected to support the highest probability for nucleation to occur at high altitude. Figure 7 shows, for the different altitude ranges previously introduced, the median condensation sink (CS) calculated from SMPS size distributions recorded at constant altitudes, i.e. apart from vertical soundings. Fig. 7. Median CS as a function of altitude range; left and right limits of the error bars stand for the 1 st and 3rd quartile, respectively. The number of data points included in the statistics is indicated on the plot for each altitude range. A more complete analysis focussed on altitudes above 2000 m was then conducted to highlight the role of the CS in the nucleation process at high altitude. Figure 8
shows
the
correlation between N5-10 particle concentration and CS, separately for the two
altitude ranges above 2000 m
.
The N5-10 show
n are 130 second averaged values coinciding
with
SMPS
measurements
used for the CS
calculation
. Based on Fig. 8, we observe that ultrafine particle concentration and CS are positively correlated, especially between 2000 and 3000 m (R2 = 0.48). The lack of measurements did not allow similar analysis at lower altitudes to compare with, but the fact that at high altitude, where the CS is usually low compared to BL stations, increased CS could favour the occurrence of nucleation has already been reported in the literature (Boulon et al., 2010; Rose et al., 2014). While lower CS values are typically
reported Fig. 8. Particle concentration in the size range 5 – 10 nm as a function of condensation sink (CS) for altitude ranges above 2000 m. Fig. 9. Median temperature and relative humidity (RH) as a function of altitude range; left and right limits of the error bars stand for the 1st and 3rd quartile, respectively. on event days compared to non-event days at BL sites, increased CS are found on event days at high altitude stations (Manninen et al., 2010). We may hypothesis that some gaseous compounds are transported, together with the pre-existing particles, from lower altitudes, and that they may be further oxidized to more condensable species involved in the nucleation process. Other parameters, such as temperature and relative humidity (RH) were previously reported to influence the nucleation process. While low temperatures were found to favor nucleation (Young et al., 2007), the role of RH seems to be more ambiguous. In fact, nucleation is likely to occur preferentially at low RH (Birmili et al., 2003), and both the nucleation rate and nucleated cluster concentration are reported to be anti-correlated with RH (Jeong et al., 2004; Sihto et al., 2006). However, nucleation events have been detected in the vicinity of clouds, where high RH are found (Clarke et al., 1998). Another aspect to consider is that among high altitude air masses, increased RH would also be associated to intrusions from the BL and hence more gaseous precursors and higher CS. Statistics concerning temperature and RH recorded during the studied flights are presented as a function of altitude range on Fig. 9. It is very clear that temperature is decreasing with altitude, especially above 3000 m where most of the temperatures are found to be negative. The same trend is observed for RH, but with higher variability. N5-10 concentrations were also directly considered as a function of temperature, RH and humidity mixing ratio (g kg-1), but the correlations between these meteorological parameters and the particle concentration was weak at all altitudes (|R2|<0.2). Global radiation, which is expected to be more intense at high altitude, and thus favor photochemical processes, including oxidation of gaseous precursors involved in the nucleation process, could also give additional explanation. We have shown so far that above Mediterranean Sea, nucleation was observed over large areas and could be favoured at high altitude, since particles in the size range 5-10 : nm are mostly seen above 1000 m. The purpose of the next section is to investigate the growth of these particles to larger diameters at high altitudes by analysing the shape of the SMPS size distributions.
IV.3.b. Investigation of particle growth at high altitude i. comparable for the two altitude ranges (Fig. 7), a faster growth at lower altitudes supported by a larger pool of condensable species could explain the differences observed on the size distributions. At night (17:00 – 05:00 UTC), the contributions of nucleation and Aitken modes to the total particle concentration are very similar between 2000 and 3000 m, being around 34%, whereas above 3000 m the nucleation mode remains dominant (46% against 36% for the Aitken mode). Again, this observation suggests that particle growth could get slower with increasing altitudes. In the morning (05:00 – 11:00 UTC), the average size distribution above 3000 m displays a nucleation mode which diameter is similar to the diameter observed at night (~ 23 nm), and with a contribution to the total particle concentration which is significantly higher than the contribution of the Aitken mode (60 and 23%, respectively). In contrast, between 2000 and 3000 m, the nucleation mode displays a large diameter (30.5 nm) and a contribution which is lower than the contribution of the Aitken mode (30 and 51%, respectively). We may hypothesis that the small particles which are seen in the nucleation mode above 3000 m do not originate from nucleation events initiated in the morning, but were rather formed the day before and are still undergoing a very slow growth process. The low concentration of nucleation mode particles between 2000 and 3000 m might be explained by a faster growth for the particles formed the day before, which have thus reached larger diameters, and also by the fact that particles nucleated in the morning (the nucleation probability between 2000 and 3000 m during the time period 05:00 - 11:00 UTC is 22%) may not have already reached the lower detection limit of the SMPS.
ii. Origin of night time nanoparticles
As previously mentioned, nanoparticles in the size range 5 – 10 nm were detected during night time (17:00 – 05:00 UTC). The of this last section is to investigate their origin, and more particularly to examine the possibility for the nigh time particles to originate from nucleation events triggered earlier during day time. The time interval 09:00 – 12:00 UTC, during which the formation of cluster particles is most frequently observed (Rose et al., 2014; 2015), was considered as a reference nucleation period, and only the significant 130 second averaged N5-10 concentrations obtained between 09:00 and 12:00 UTC were further considered. We estimated the half-life time of these particles, i.e. the time tfin at which these concentrations reached half of their initial values, obtained at t0, under the following assumptions: 1) particles are removed from the size range 5 – 10 nm by coagulation and growth processes and 2) the coagulation sink calculated at t0 is constant during particle growth. tfin is finally given by Eq. (2):
t fin t 0 0.5 f Coag * GR 5 (2)
Where Coag is the coagulation sink of 5 nm particles derived from SMPS data, and GR is the particle growth rate. The factor f represents the fraction of the aerosol population which is activated for growth, and was assumed to be equal to unity. Particle GRs were estimated from the average shift of the nucleation mode diameter observed on the SMPS size distributions between night time (17:00 – 05:00 UTC) and morning hours (05:00 – 11:00 UTC) for the altitude range 2000 – 3000 m (Table A1, Fig. 10). An average value of 0.31 nm h-1 was found. However, GR was previously reported to increase with particle size (Yli-Juuti et al., 2011; Kulmala et al., 2013), this is the reason why lower GR of 0.1 nm h-1 was also used to describe particle growth in the size range 5 – 10 nm. In order to complete our sensitivity study regarding the GR, additional higher value of 1 nm h-1 was selected. The results of this analysis are reported on Fig. 11, which first indicates that particle life time increases with altitude, which might be explained by decreasing coagulation sinks. Considering the scenario with intermediate GR of 0.3 nm h -1, initial N5-10 concentrations are all divided by two before night time, suggesting that, considering the relatively high concentrations found at night, there would be an additional source of 5-10 nm particles at high altitudes. The same conclusion is obtained with GR = 1 nm h -1. IV.4. Conclusion
We investigated the occurrence of nucleation events above the Mediterranean Sea using data obtained during research flights performed in the framework of the HYMEX Fig. 11. Estimations of tfin, the time at which N5-10 concentrations reach half of their initial values, as a function of altitude and particle growth rate. September 2012. Based on our observations, nucleation takes place over large areas above the Mediterranean Sea in all air mass types. Nucleation probability slightly varies with air mass origin, but the signature of the different air mass types is however complex to distinguish in terms of N5-10 concentrations. In Western Europe and South Mediterranean Sea flows, more frequently observed compared to other air mass types, maximum concentrations were obtained for fetches around 5 hours, but significant concentrations persist over fetches up to 60 hours, suggesting that the nucleation process could be more influenced by parameters inherent of the marine troposphere. The analysis of the vertical extension of nucleation showed that the process was promoted at high altitude, above 1000 m, and especially between 2000 and 3000 m. Simultaneous analysis of the boundary layer height indicated that these altitudes often corresponded to free tropospheric conditions. Vertical gradient of the condensation sink, together with temperature and humidity might explain the increasing nucleation probability with altitude. However, for a given high altitude range, larger N5-10 concentrations were found for larger CS, likely associated with higher oxidation of gas phase precursors. The investigation of the global shape of the particle size distributions derived from SMPS measurements finally allowed us to study the particle growth above 2000 m, and more particularly the relative contribution of the different particle modes to the total concentration as a function of time and altitude. After they formed, particles appear to grow to larger sizes above 2000 m, reaching the Aitken mode range, but with growth rates which seem to decrease with altitude. This slow growth, coupled with low coagulation sinks, may favour longer subsistence for nanoparticles in the size range 5 – 10 nm, and could explain the detection of these small particles during night time, several hours after their formation. Our findings demonstrate that high altitude could promote the occurrence NPF, not only over continental areas, as previously suggested by Boulon et al. (2011), but also over open seas, with indications of marine precursors. This result supports the model study by Makkonen et al. (2012), which predicts that nucleation could have a significant contribution to the cloud condensation nuclei (CCN) concentration over the Mediterranean Sea, and indicates that this contribution could be even more decisive at high altitude, where clouds form. Appendix A Table A1. Parameters of the Gaussians used to fit the SMPS size distributions as a function of daytime for the altitude ranges above 2000 m. 05:00 – 11:00 Mode M1 Parameters 2000 – 3000 m >3000 m N (cm-3) 126.3 ± 70.4 200.3 ± 117.9 1.32 ± 0.01 1.34 ± 0.03 292 Annexes : Publications M2 M3 M4 dp (nm) 30.5 ± 3.0 23.5 ± 3.7 N (cm-3) 211.3 ± 130.1 78.1 ± 62.5 1.36 ± 0.02 1.37 ± 0.01 dp (nm) 51.6 ± 5.1 46.1 ± 7.6 N (cm-3) 130.4 ± 88.0 49.1 ± 23.8 1.37 ± 0 1.37 ± 0 dp (nm) 99.7 ± 9.5 93.7 ± 8.7 N (cm-3) 30.9 ± 26.9 9.6 ± 9.2 1.35 ± 0.05 1.31 ± 0.03 dp (nm) 205.4 ± 13.7 205.0 ± 9.7 Parameters 2000 – 3000 m >3000 m N (cm-3) 248.1 ± 130.0 155.1 ± 126.0 1.32 ± 0.02 1.34 ± 0.03 dp (nm) 28.6 ± 3.6 24.6 ± 4.8 N (cm-3) 161.8 ± 174.4 83.5 ± 46.9 1.36 ± 0.02 1.37 ± 0.01 dp (nm) 52.4 ± 5.8 49.7 ± 7.4 N (cm-3) 150.8 ± 124.7 67.4 ± 36.0 1.38 ± 0.01 1.37 ± 0.01 dp (nm) 99.7 ± 8.9 97.5 ± 10.0 N (cm-3) 39.0 ± 38.6 17.7 ± 19.4 1.35 ± 0.04 1.32 ± 0.04 dp (nm) 201.9 ± 12.7 224.3 ± 48.4 11:00 – 17:00 Mode M1 M2 M3 M4 293 Annexes : Publications 17:00 – 05:00 Mode M1 M2 M3 M4 Parameters 2000 – 3000 m >3000 m N (cm-3) 224.9 ± 156.7 176.6 ± 117.7 1.32 ± 0.02 1.33 ± 0.03 dp (nm) 27.7 ± 3.1 23.8 ± 3.5 N (cm-3) 227.2 ± 169.4 136.9 ± 9.9 1.37 ± 0 1.37 ± 0 p (nm) 50.6 ± 4.1 46.4 ± 5.6 N (cm-3) 167.3 ± 108.7 61.4 ± 58.4 1.36 ± 0.01 1.35 ± 0.02 dp (nm) 101.6 ± 7.7 102.8 ± 8.4 N (cm-3) 44.7 ± 40.7 9.2 ± 6.2 1.37 ± 0.05 1.39 ± 0.04 dp (nm) 216.4 ± 17.9 240.3 ± 12.1
Acknowledgement
HyMeX SOP1 was supported by CNRS, Météo-France, CNES, IRSTEA, INRA through the large interdisciplinary international program MISTRALS (Mediterranean Integrated STudies at Regional And Local Scales) dedicated to the understanding of the Mediterranean Basin environmental process (http://www.mistrals-home.org). We also would like to thank SAFIRE for their support during instrument integration on the French ATR-42 and HYMEX SOP1 campaign execution. figures Figure 2-1 Classifications usuelles pour l'étude de la taille des particules d'aérosol. 17 Figure 2-2 Schéma simplifié en deux étapes du processus de formation de nouvelles particules 20 Figure 2-3
Pollution et santé : une problématique mondiale. a) Nombre de jours d'espérance de vie perdus en Europe par exposition aux PM2.5 en 2005 ; b) Les taux de pollution record à Pékin font régulièrement la une de l'actualité (Trafic à Pékin le 23 janvier 2013, photo Jason Lee, Reuters ; Port du masque le 2 mai 2013, photo Kim Kyung Hoon, Reuters.).
23 Figure 2-4 Principaux forçages impactant le bilan radiatif terrestre (GIEC, 2013)
26 Figure 2-5 Impacts conjoints 1) des politiques de réduction des émissions de particules et de SO2 et 2) de l'augmentation des concentrations en BCOV et DMS due au réchauffement climatique sur le força
ge
total li
é
aux
particules d'aérosol. 28 Figure 2-6 Variation de l'énergie libre ΔG associée à la formation d'un embryon en fonction du diamètre de l'embryon. 30 Figure 2-7 Schéma des étapes de croissance d'un embryon impliquant des composés organiques. 43 Figure 2-8 Mécanismes de réaction envisagés entre les alcènes et l'acide sulfurique (Zhang and Wexler, 2002). 44 Figure 2-9 Evolution du rapport de saturation nécessaire à la croissance d'un embryon en fonction du diamètre de l'embryon, avec et sans prise en compte du phénomène d'adsorption. 45 Figure 3-1 Schéma de fonctionnement d'un DMA cylindrique. 53 Figure 3-2 Schéma de fonctionnement de l'AIS et du NAIS. 54 Figure 3-3 Schéma de fonctionnement du PSM 57 Figure 3-4 Illustration des différents types d'évènements : a) type Ia, b) type Ib, c) type II et d) bump 60 Figure 3-5 Détermination du GR par la méthode des maxima. a)
Détermination du temps d'obtention des maxima de concentration dans chaque canal de taille. b) Régression linéaire sur les couples temps-diamètre dont la pente correspond au GR. c) Visualisation de la croissance des
particul
es dans la première classe de
taille sur le spectre dimensionnel. 62 Figure 4-1 Fréquences mensuelles moyennes d'obtention de jours avec ou sans évènement de FNP et de jours indéfinis au PDD entre Février 2007 et Février 2012. 68 Figure 4-2 Evolution de la température (figure supérieure) et de la concentration en carbone suie (figure inférieure) durant la période d'étude. L'absence de valeurs de BC durant la période 3 est due à une panne instrumentale. Les lignes pointillées et les valeurs chiffrées associées apposées sur la figure correspondent aux moyennes calculées pour les 3 sous périodes 1, 2 et 3.
70 Figure 4-3 Hauteur de la CLA dérivée des mesures LIDAR (points noirs). La ligne pointillée rouge correspond à l'altitude du PDD. 71 Figure 4-4 Variations moyennes des taux de formation chargés et total pour les embryons de 1.5 et 3 nm de diamètre. Seules les heures de jour sont représentées sur la figure. Figure 4-5 Exemples d'évènements de FNP multiples a) consécutifs le 6 Mai 2012 et b) séparés le 22 Mars 2012 à Chacaltaya. 75 Figure 4-6 Fréquences mensuelles d'observation d'évènements de FNP en fonction du nombre d'évènements observés par jour. 76 Figure 4-7 Variations mensuelles du type d'évènement identifié pour les évènements simples et les évènements placés en première position dans le cas d'évènements multiples. Le nombre total de jours d'évènements détectés chaque mois est donné en haut de la figure. 76 Figure 4-8 Taux de formation médians a) des particules totales et b) chargées de 2 nm observés pour les évènements simples et les évènements placés en première position lors d'évènements multiples. Les extrémités inférieure et supérieure des barres d'erreurs correspondent au premier et troisième quartile, respectivement. Le nombre d'évènements inclus dans les statistiques est indiqué pour chaque mois en haut de la figure. 77 Figure 4-9 IIN médians ou moyens calculés pour des stations d'altitude représentés en fonction de l'altitude. 79 Figure 4-10 Variations mensuelles du taux de croissance médian des particules pour les trois intervalles de diamètres 1.5-3 nm, 3-7 nm et 7-20 nm pour les évènements simples et les évènements placés en première position lors d'évènements multiples. Les extrémités inférieure et sup
des barres d'erreurs correspondent au premier et troisième quartile, respectivement.
80 Figure 4-11 Représentation de l'ensemble des trajectoires des vols réalisés entre le 11 Septembre et le 5 Novembre 2012 durant la campagne HYMEX. 85 Figure 4-12 Méthode de filtrage pour la détection visuelle des points de mesure obtenus audessus de la terre. Les points bleus sont situés une distance de 1000±50 m de la position de l'avion. 86 Figure 4-13 Statistiques concernant la détection de concentration a) N5-10 et b) N10-20 significatives pour différents créneaux d'altitude et différentes plages horaires. Le nombre total de points de mesure correspondant à chaque couple heure/altitude est indiqué sur la droite de chacune des figures. 86
Figure 4-14 Profils de concentration N5-10 obtenus lors des sondages verticaux effectués par l'avion au-dessus de la mer. Les points noirs corespondent aux concentrations inférieures au seuil de 175 cm-3 et les concentrations supérieures au seuil sont indiquées par l'échelle de couleur. Les étoiles roses indiquent la hauteur de couche limite obtenues par ECMWF. Les profils obtenus au cours de phases d'atterissage ou de décollage sont repérés par un carré noir 87 Figure 4-15 Illustration de la méthode de classification des masses d'air échantillonnées le long de la trajectoire de vol (points noirs). Les lignes rouges délimitent les différents secteurs géographiques définis pour la classification. 89 Figure 4-16 Reconstruction des distributions en taille de particules mesurées par le SMPS à partir des paramètres moyens des modes obtenus lors de l'analyse indiciduelle de chaque distribution en fonction de l'altitude et de l'heure. 91 Figure 4-17 Evolution de la concentration du mode nucléation en fonction de l'heure et de l'altitude. 93 Figure 4-18 Evolution du diamètre du mode nucléation en fonction de l'heure et de l'altitude. 93
296 Index des figures
Figure 4-19 Localisation des valeurs de N5-10 significatives détectées durant HYMEX en fonction de l'origine des masses d'air. Les valeurs de N5-10 sont indiquées par l'échelle de couleur. 94 Figure 4-20 Spectres dimensionnels des particules mesurés par SMPS aux stations de Ersa et Cap Es Pinar sur l'ensemble de la campagne de mesure. Les épisodes de FNP identifiés sont indiqués sur les spectres. 95 Figure 4-21 Rétro trajectoires des masses d'air ayant atteint les deux sites de mesure au cours de la première période de F
observ
ée entre le 04 et le 09 Juillet.
97 Figure 4-22 Rétro trajectoires des masses d'air ayant atteint les deux sites de mesure au cours de la deuxième période de FNP observé entre le 27 Juillet et le 03 Août. 98 Figure 4-23 Rétro trajectoires des masses d'air ayant atteint les deux sites de mesure au cours de la troisième période de FNP observé entre le 09 et le 12 Août. 99 Figure 4-24 Concentration en particules de diamètre compris entre 3 et 10 nm (N3-10) sur la trajectoire du vol 51. Seules les valeurs de N3-10 au-dessus du seuil sont représentées, et indiquées par l'échelle de couleur. Les points de petite taille correspondent à la portion de vol réalisée à basse altitude (~215 m) alors que les points de grosse taille indiquent une altitude supérieure (~3400 m).
100 Figure 4-25 Distributions en taille de particules mesurées par SMPS le 30 Juillet durant les périodes sélectionnées du vol 51. La couleur des distributions est liée à la position de l'avion, indiquée sur la Figure 4-27. 101 Figure 4-26 En traits pleins : rétro trajectoires des masses d'air échantillonnées le long de la trajectoire de l'avion toutes les dix minutes (représentée en noir) ; en traits pontillés : retro trajectoires des masses d'air ayant atteint la station dans le même temps, à 13h00, 14h00 et 15h00 101 Figure 4-27 Code couleur indiquant la position de l'avion lors de l'obtention des distributions en taille présentées sur la Figure 4-26. 102 Figure 4-28 Diamètre et concentration des modes représentatifs des distributions en taille mesurées par SMPS durant le vol 51 du 30 Juillet. 102 Figure 4-29 Concentration en particules de diamètre compris entre 3 et 10 nm (N3-10) sur la trajectoire du vol 52. Seules les valeurs de N3-10 au-dessus du seuil sont représentées, et indiquées par l'échelle de couleur 103 Figure 4-30 En traits pleins : rétro trajectoires des masses d'air échantillonnées le long de la trajectoire de l'avion toutes les dix minutes (représentée en noir) ; en traits pointillés : retro trajectoires des masses d'air ayant atteint la station dans le même temps, à 13h00, 14h00 et 15h00 104 Figure 4-31 Distributions en taille de particules mesurées par SMPS le 30 Juillet durant les périodes sélectionnées du vol 52. La couleur des distributions est liée à la position de l'avion, indiquée sur la Figure 4-32. 104 Figure 4-32 couleur indiquant la position de l'avion lors de l'obtention des distributions en taille présentées sur la Figure 4-31. 105 Figure 4-33 Diamètre et concentration des modes représentatifs des distributions en taille mesurées par SMPS durant le vol 52 du 1er Août. 105 Figure 4-34 Rapport des valeurs des paramètres des modes obtenus à bord de l'ATR-42 sur les valeurs obtenues au sol. 106 297 Index des figures Figure 4-35 Evolution temporelle des concentrations en particules de diamètre compris entre 11 et 15 nm (N11-15), 15 et 20 nm (N15-20) et 20 et 25 nm (N20-25) pour la journée du 05 Juillet. 107 Figure 4-36 Evolution des distributions en taille des particules mesurées par le SMPS et moyennées sur 1h entre 08h00 et 18h00 aux deux stations (05 Juillet). 108 Figure 4-37 Evolution temporelle des paramètres des modes (diamètre, concentration) et de leur contribution relative (05 Juillet). 109 Figure 4-38 Rétro trajectoires des masses d'air échantillonnées à Cap Es Pinar le 5 Juillet lors de l'obtention du maximum de la concentration en particules de 20 nm (tmax), 1h et 2h avant, et 1h et 2h après. Le lieu de déclenchement de la nucléation estimé à partir de GR15-25 et de la rétro trajectoire de la masse d'air échantillonnée à tmax est indiqué sur la carte 110 Figure 4-39 Rétro trajectoires des masses d'air échantillonnées à Ersa le 05 Juillet lors de l'obtention du maximum de la concentration en particules de 20 nm (tmax), 1h et 2h avant, et 1h et 2h après. Le lieu de déclenchement de la nucléation estimé à partir de GR15-25 et de la rétro trajectoire de la masse d'air échantillonnée à tmax est indiqué sur la carte 111 Figure 4-40 Evolution temporelle des concentrations en particules de diamètre compris entre 11 et 15 nm (N11-15), 15 et 20 nm (N15-20) et 20 et 25 nm (N20-25) pour la journée du 29 Juillet. 112 Figure 4-41 Evolution des distributions en taille des particules mesurées par le SMPS et moyennées sur 1h entre 08h00 et 18h00 aux deux stations (29 Juillet). 112 Figure 4-42 Evolution temporelle des paramètres des modes (diamètre, concentration) et de leur contribution relative (29 Juillet) 113 Figure 4-43 Rétro trajectoires des masses d'air échantillonnées à Cap Es Pinar le 29 Juillet lors de l'obtention du maximum de la concentration en particules de 20
nm (tmax), 1h et 2h avant, et 1h et 2h après. Le lieu de déclenchement de la nucléation estimé à partir de GR15-25 et de la rétro trajectoire de la masse d'air échantillonnée à tmax est indiqué sur la carte.
114 Figure 4-44 Rétro trajectoires des masses d'air échantillonnées à la station le 29 Juillet lors de l'obtention du maximum de la concentration en particules de 20 nm (tmax), 1h et 2h avant, et 1h et 2h après. Le lieu de déclenchement de la nucléation estimé à partir de GR15-25 et de la rétro trajectoire de la masse d'air échantillonnée à tmax est indiqué sur la carte
115
Figure 4-45 Evolution temporelle des concentrations en particules de diamètre compris entre 11 et 15 nm (N11-15), 15 et 20 nm (N15-20) et 20 et 25 nm (N20-25) pour la journée du 09 Août. 115 Figure 4-46 Evolution des distributions en taille des particules mesurées par le SMPS et moyennées sur 1h entre 08h00 et 18h00 aux deux stations (09 Août). 116 Figure 4-47 Evolution temporelle des paramètres des modes (diamètre, concentration) et de leur contribution relative (09 Août). 116 Figure 4-48 Rétro trajectoires des masses d'air échantillonnées à Cap Es Pinar le 09 Août lors de l'obtention du maximum de la concentration en particules de 20 nm (tmax), 1h et 2h avant, et 1h et 2h après. Le lieu de déclenchement de la nucléation estimé à partir de GR15-25 et de la rétro trajectoire de la masse d'air échantillonnée à tmax est indiqué sur la carte 117 Figure 4-49 Rétro trajectoires des masses d'air échantillonnées à Ersa le 09 Août lors de l'obtention du maximum de la concentration en particules de 20 nm (tmax), 1h et 2h avant, et 1h et 2h après. Le lieu de déclenchement de la nucléation estimé à partir de GR15-25 et de la rétro trajectoire de la masse d'air échantillonnée à tmax est indiqué sur la carte 118 Figure 5-1 Variations mensuelles de la concentration en embryons chargés (0.8 – 1.9 nm). La ligne rouge représente la médiane mensuelle, les extrémités inférieure et supérieure du 298 Index des figures rectangle bleu représentent le 1er et le 3ème quartile, respectivement, et les
émités des lignes noires représentent le 10eme et le 90eme percentile. 122
Figure 5-2 Variation journalière de la concentration en embryons positifs selon la saison pour les jours a) avec évènement de FNP et b) sans évènement. La ligne rouge représente la médiane mensuelle, les extrémités inférieure et supérieure du rectangle bleu représentent le 1er et le 3ème quartile, respectivement, et les extrémités des lignes noires représentent le 10eme et le 90eme percentile. 123 Figure 5-3 Résultat du test de Mann – Whitney appliqué aux concentrations en embryons positifs mesurées les jours avec et sans évènement de FNP séparément pour chaque saison. Les carrés sont coloriés lorsque pour l'heure correspondante l'hypothèse nulle d'échantillons présentant des médianes différentes ne peut être rejetée avec un seuil de 5%.
124 Figure 5-4 Résultat du test de Mann – Whitney appliqué aux concentrations en embryons négatifs mesurées les jours avec et sans évènement de FNP séparément pour chaque saison. Les carrés sont coloriés lorsque pour l'heure correspondante l'hypothèse nulle d'échantillons présentant des médianes différentes ne peut être rejetée avec un seuil de 5%. 125 Figure 5-5 Variation journalière du diamètre du mode des embryons positifs selon la saison pour les jours a) avec évènement de FNP et b) sans évènement. La ligne rouge représente la médiane mensuelle, les extrémités inférieure et supérieure du rectangle bleu représentent le 1er et le 3ème quartile, respectivement, et les extrémités des lignes noires représentent le 10eme et le 90eme percentile. 126 Figure 5-6 Résultat du test de Mann – Whitney appliqué aux diamètres du mode des embryons positifs obtenus les jours avec et sans évènement de FNP séparément pour chaque saison. Les carrés sont coloriés lorsque pour l'heure correspondante l'hypothèse nulle d'échantillons présentant des médianes différentes ne peut être rejetée avec un seuil de 5%. 126 Figure 5-7 Résultat du test de Mann – Whitney appliqué aux diamètres du mode des embryons négatifs obtenus les jours avec et sans évènement de FNP séparément pour chaque saison. Les carrés sont coloriés lorsque pour l'heure correspondante l'hypothèse nulle d'échantillons présentant des médianes différentes ne peut être rejetée avec un seuil de 5%. 127 Figure 5-8 Taux d'ionisation des embryons positifs dérivé de l'équation régissant l'évolution de la concentration
embryons positifs (Qcalc) en fonction du taux d'ionisation calculé à partir de la concentration en 222Rn et du rayonnement externe (Qmes). Différents symboles sont utilisés pour les jours avec et sans évènement de FNP et la couleur des marqueurs correspond à l'heure de la mesure.
132 Figure 5-9 Variation journalière des concentrations en embryons neutres et chargés pour les jours avec et sans évènements de FNP. 134 Figure 5-10 Illustration de la corrélation entre les concentrations en acide sulfurique mesurées et les valeurs issues du proxy ajusté sur les mesures. 137 Figure 5-11 Comparaison des concentrations en acide sulfurique issues des mesures de l'ApiToF et des différents proxys. 138 Figure 5-12 Evolution de la concentration en acide sulfurique durant la période d'étude. Les valeurs de BC manquantes sont dues à une panne instrumentale. Les lignes pointillées et les 299 Index des figures valeurs chiffrées associées apposées sur la figure correspondent aux moyennes calculées pour les 3 sous périodes 1, 2 et 3.
139 Figure 5-13 Correspondance entre la concentration en embryons et la concentration en acide sulfurique pour les jours avec et sans évènement de FNP. 140 Figure 5-14 a) Fréquence de FNP et b) nombre d'évènements de FNP détectés par jour en fonction du parcours des masses d'air. L'étoile marque la position du site de mesure et les lignes noires délimitent les différents secteurs utilisés pour l'étude de l'origine des masses d'air (océanique, continentale, Amazonienne et locale). 142 Figure 5-15 Lieu d'initiation du processus de FNP dans les masses d'air océaniques avant détection à la station de mesure. 143 Figure 5-16 Taux de formation des particules de 2 nm : a) total, b) positif et c) négatif. 144 Figure 5-17 Taux de croissance des particules sur la gamme de diamètre a) 1 – 3 nm, b) 3 – 7 nm et c) 7 – 20 nm. 145 Figure 5-18 Evolution de la concentration en chlorophylle-a dans chaque mésocosme. 146 Figure 5-19 Dispositif instrumental déployé pour la mesure in-situ des propriétés de l'atmosphère de chaque mésocosme 146 Figure 5-20 Evolution de la concentration totale en particules mesurée par le SMPS (14 – 660nm), de la concentration en particules comprises entre 1 et 14nm et de la concentration en iode en phase particulaire. Afin de faciliter la lecture de la figure, la même limite supérieure a été choisie pour les trois axes verticaux et les valeurs supérieures à cette limite ont été indiquées pour la figure centrale. Les valeurs présentées sont des valeurs moyennes calculées entre 07h00 h00,
.
147 Figure 5-21 Corrélation entre la concentration en iode et la concentration en particules dans le mésocosme B pour la journée du 19 Mai entre 07h00 et 19h00 148 Figure 5-22 Comparaison des valeurs de CS obtenues au cours des études menées dans le cadre de cette thèse avec les valeurs issues des travaux de Manninen et al. (2010) concernant des sites caractérisés par des altitudes et des environnements variés. Pour les résultats de la campagne HYMEX, il est indiqué la plage d'altitude à laquelle correspond la valeur. En outre, toutes les valeurs indiquées sont des médianes, à l'exception des valeurs concernant l'étude du PDD en FT qui sont des moyennes.
150 Figure 5-23 Corrélation entre la concentration en embryons et la valeur du puits de condensation en fonction de la charge des embryons et de l'occurrence ou non d'un évènement de FNP. 151 Figure 5-24 Variations du CS médian avec l'altitude dans les masses d'air d'ouest. Les extrémités inférieure et supérieure des barres d'erreur représentent respectivement le 1er et le 3ème quartile. 152 Figure 5-25 Reconstruction des spectres SMPS à partir des paramètres des 4 modes identifiés pour chaque spectre dans les masses d'air d'ouest au-dessus de 2000 m. La couleur des spectres indique le temps passé par la masse d'air au-dessus de la mer avant d'avoir été échantillonnée au niveau de l'avion. 153 Figure 5-26 Concentration en particules de diamètre compris entre a) 5 et 10 nm et b) 10 et 20 nm en fonction du temps passé par la masse d'air au-dessus de la mer avant échantillonnage par l'avion. Les points bleus correspondent aux spectres présentant un mode nucléation dominant sur la Figure 5-25.
des
figures Figure 6-1 Illustration de la classification des évènements de type I en fonction de leur contribution à la formation de CCN. Jusqu'à 20 nm, le spectre dimensionnel est issu des mesures du NAIS ; pour les diamètres supérieur il est issu des mesures du SMPS. a) On observe une contribution claire de la FNP à l'augmentation de la concentration en particules pour les diamètres excédant 100 nm ; b) La croissance des particules n'excède pas 50 nm ; c) La FNP ne contribue pas à la formation des CCN (le premier diamètre seuil de 50 nm n
est pas atteint) ; d) Il est difficile de distinguer la contribution de l'évènement du jour étudié, de celle de l'évènement du jour précédent : jour classé comme indéfini. Les lignes noires représentent les trois diamètres d'activation choisis dans cette étude : 50 nm, 80 nm et 100 nm.
158 Figure 6-2 Détermination des temps et des concentrations de référence utilisés pour le calcul des augmentations absolue et relative des concentrations en CCN dues à la FNP. Participation à des campagnes de mesure durant la thèse Campagne de mesure intensive au puy de Dôme, avec en particulier le déploiement de l'AMS communautaire et d'un Api-Tof pour la caractérisation de la composition chimique des embryons chargés (Janvier – Février 2012) ; Expérience sur la nucléation de COV biogéniques dans CESAM (Mars 2012) ; Projet PIREP pour l'étude de la nucléation dans les échappements d'un moteur diesel, en aval d'un filtre à particules (Mai 2012) ; Projet Medsea pour la caractérisation en mésocosmes des échanges mer-atmosphère dans le cadre de l'acidification des océans, Calvi, Corse (Juin 2012) ; Projet SAM pour l'étude en chambre de simulation (Lyon, Novembre – Décembre 2012) et en atmosphère semi controlée (mésocosmes, Calvi, Mai 2013) de la nucléation en atmosphère marine dans le cadre d'une augmentation artificielle de l'activité biologique de l'eau. Participation à des workshop/école d'été Ecole d'été : Formation and growth of atmospheric aerosols, Hyytiälä, Finlande, Août 2012; PSM user meeting, organisé par la société AIRMODUS, Hyytiälä, Finlande, Août 2012; Workshop sur la nucléation et la formation de nouvelles particules, Hyytiälä, Finlande, Août 2012 (présentation d'un poster); ACTRIS WP3 Technical Meeting, Leipzig, Allemagne, Octobre 2012; ACTRIS 4th General Meeting, Clermont-Ferrand, Juin 2014 (presentation d'un poster). Liste des publications E. Asmi, E. Freney, M. Hervo, D. Picard, C. Rose, A. Colomb and K. Sellegri: Aerosol Cloud Activation in summer and winter at puy de Dôme high altitude site in France, Atmos. Chem. Phys., C. Rose, J. Boulon, M. Hervo, H. Holmgren, E. Asmi, M. Ramonet, P. Laj and K. Sellegri: Long-term observations of cluster ion concentration, sources and sinks in clear sky conditions at the high-altitude site of the puy de Dôme, France, Atmos. Chem. Phys., 13, 11573-11594, 2013. H. Holmgren, K.Sellegri, M. Hervo, C. Rose, E. Freney, P.Villani and P. Laj: Hygroscopic properties and mixing state of aerosol measured at the high altitude site puy de Dôme (1465 m a.s.l.), France, Atmos. Chem. Phys., 14, 9537-9554, 2014. C. Rose, K. Sellegri, E. Asmi, M. Hervo, E. Freney, H. Junninen, J. Duplissy, M. Sipilä, J. Kontkanen, K. Lehtipalo and M. Kulmala: Major contribution of neutral clusters to new particle formation in the free troposphere, Atmos. Chem. Phys. Discuss., 14, 18355-18388, 2014. A. N. Schwier, C. Rose, E. Asmi, A.M. Ebling, W.M. Landing, S. Marro, M.-L. Pedrotti, A. Sallon, F. Iuculano, S. Agusti, A. Tsiola, P. Pitta, J. Louis, C. Guieu, F. Gazeau and K. Sellegri: Primary marine aerosol emissions from the Mediterranean Sea during pre-bloom and oligotrophic conditions: correlations to seawater chlorophyll-a from a mesocosm study, Atmos. Chem. Phys. Discuss., 14, 26187–26230, 2014. C. Rose, K. Sellegri, F. Velarde, I. Moreno, M. Ramonet, K. Weinhold, M. Andrade, A. Wiedensohler and P. Laj: Multiple daytime nucleation events at the high altitude station of Chacaltaya (5240 m a.s.l.), Bolivia, Atmos. Environ.
102, 18-29, 2015. Activités de vulgarisation scientifique Monitorat; Visite de la station de mesure du puy de Dôme pour des scolaires et dans le cadre de la Fête de la science pour un large public ; Présentation brève des activités de l'observatoire pour l'émission télévisée « Chroniques d'en haut ». La formation de nouvelles particules est un processus complexe à l'origine d'une fraction importante des concentrations en nombre de particules observées dans l'atmosphère. En jouant le rôle de noyau de condensation (CCN) pour la formation des gouttelettes de nuage, les particules issues de ce processus impactent le bilan radiatif terrestre. Fréquemment observée et documentée à basse altitude, la formation de nouvelles particules a plus rarement fait l'objet d'études à haute altitude. L'analyse des données obtenues en 2012 à la plus haute station du monde, Chacaltaya (5240 m, Bolivie) révèle une fréquence d'observation annuelle du processus remarquablement élevée (64%), avec de nombreux évènements multiples. Les mesures conduites à la station du puy de Dôme (1465 m, ACTRIS, GAW) qui bénéficie d'un dispositif instrumental rare ont plus particulièrement permis de mettre en évidence le déroulement du processus en troposphère libre. Une analyse complète de l'extension verticale du processus rendue possible grâce aux données aéroportées obtenues au-dessus du bassin Méditerranéen dans le cadre du projet HYMEX (MISTRALS, automne 2012) a montré qu'en plus d'être observé à haute altitude, le processus de formation de nouvelles particules semblait y être clairement favorisé, avec une probabilité d'observation multipliée par 10 au-dessus de 1000 m. De plus, à ces altitudes le processus de formation de nouvelles particules pourrait être une source importante de CCN, comme le suggèrent les résultats obtenus à Chacaltaya, où dans 68% des évènements analysés les particules formées atteignent des diamètres suffisants pour jouer le rôle de CCN. La diversité des environnements associés aux bases de données utilisées a également permis d'apporter des éléments relatifs à la compréhension du processus point de vue de la charge des embryons formés, de l'identité des précurseurs gazeux impliqués et des paramètres atmosphériques influençant le processus. Ces éléments sont déterminants pour une prise en compte optimale du processus de formation de nouvelles particules dans les modèles. Mots-clés: aérosol, nucléation, croissance, altitude, précurseur.
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