Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Outros Autores: | , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.18/2752 |
Resumo: | This study investigated the levels of DNA strand breaks and formamidopyrimidine DNA glycosylase (FPG) sensitive sites, as assessed by the comet assay, in peripheral blood mononuclear cells (PBMC) from healthy women from five different countries in Europe. The laboratory in each country (referred to as 'centre') collected and cryopreserved PBMC samples from three donors, using a standardised cell isolation protocol. The samples were analysed in 13 different laboratories for DNA damage, which is measured by the comet assay. The study aim was to assess variation in DNA damage in PBMC samples that were collected in the same way and processed using the same blood isolation procedure. The inter-laboratory variation was the prominent contributor to the overall variation. The inter-laboratory coefficient of variation decreased for both DNA strand breaks (from 68 to 26%) and FPG sensitive sites (from 57 to 12%) by standardisation of the primary comet assay endpoint with calibration curve samples. The level of DNA strand breaks in the samples from two of the centres (0.56-0.61 lesions/10(6) bp) was significantly higher compared with the other three centres (0.41-0.45 lesions/10(6) bp). In contrast, there was no difference between the levels of FPG sensitive sites in PBMC samples from healthy donors in the different centres (0.41-0.52 lesion/10(6) bp). |
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Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratoriesDNA DamageComet AssayEnvironmental Cancer RiskAr e Saúde OcupacionalGenotoxicidade AmbientalThis study investigated the levels of DNA strand breaks and formamidopyrimidine DNA glycosylase (FPG) sensitive sites, as assessed by the comet assay, in peripheral blood mononuclear cells (PBMC) from healthy women from five different countries in Europe. The laboratory in each country (referred to as 'centre') collected and cryopreserved PBMC samples from three donors, using a standardised cell isolation protocol. The samples were analysed in 13 different laboratories for DNA damage, which is measured by the comet assay. The study aim was to assess variation in DNA damage in PBMC samples that were collected in the same way and processed using the same blood isolation procedure. The inter-laboratory variation was the prominent contributor to the overall variation. The inter-laboratory coefficient of variation decreased for both DNA strand breaks (from 68 to 26%) and FPG sensitive sites (from 57 to 12%) by standardisation of the primary comet assay endpoint with calibration curve samples. The level of DNA strand breaks in the samples from two of the centres (0.56-0.61 lesions/10(6) bp) was significantly higher compared with the other three centres (0.41-0.45 lesions/10(6) bp). In contrast, there was no difference between the levels of FPG sensitive sites in PBMC samples from healthy donors in the different centres (0.41-0.52 lesion/10(6) bp).Environmental Cancer Risk, Nutrition and Individual Susceptibility 2 (ECNIS2) towards the establishment of a virtual European Centre for Research and Education on Cancer, Environment and Food, European Commission FP7-KBBE-2010–4 (Contract No. 266198).Oxford University Press/UK Environmental Mutagen SocietyRepositório Científico do Instituto Nacional de SaúdeGodschalk, R.W.Ersson, C.Stępnik, M.Ferlińska, M.Palus, J.Teixeira, João PauloCosta, S.Jones, G.D.Higgins, J.A.Kain, J.Möller, L.Forchhammer, L.Loft, S.Lorenzo, Y.Collins, A.R.van Schooten, F.J.Laffon, B.Valdiglesias, V.Cooke, M.Mistry, V.Karbaschi, M.Phillips, D.H.Sozeri, O.Routledge, M.N.Nelson-Smith, K.Riso, P.Porrini, M.López de Cerain, A.Azqueta, A.Matullo, G.Allione, A.Møller, P.2015-02-03T13:10:07Z2014-072014-07-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.18/2752engMutagenesis. 2014 Jul;29(4):241-9. doi: 10.1093/mutage/geu012. Epub 2014 Apr 15.0267-835710.1093/mutage/geu012info:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-20T15:39:24Zoai:repositorio.insa.pt:10400.18/2752Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:37:39.330142Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories |
title |
Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories |
spellingShingle |
Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories Godschalk, R.W. DNA Damage Comet Assay Environmental Cancer Risk Ar e Saúde Ocupacional Genotoxicidade Ambiental |
title_short |
Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories |
title_full |
Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories |
title_fullStr |
Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories |
title_full_unstemmed |
Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories |
title_sort |
Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories |
author |
Godschalk, R.W. |
author_facet |
Godschalk, R.W. Ersson, C. Stępnik, M. Ferlińska, M. Palus, J. Teixeira, João Paulo Costa, S. Jones, G.D. Higgins, J.A. Kain, J. Möller, L. Forchhammer, L. Loft, S. Lorenzo, Y. Collins, A.R. van Schooten, F.J. Laffon, B. Valdiglesias, V. Cooke, M. Mistry, V. Karbaschi, M. Phillips, D.H. Sozeri, O. Routledge, M.N. Nelson-Smith, K. Riso, P. Porrini, M. López de Cerain, A. Azqueta, A. Matullo, G. Allione, A. Møller, P. |
author_role |
author |
author2 |
Ersson, C. Stępnik, M. Ferlińska, M. Palus, J. Teixeira, João Paulo Costa, S. Jones, G.D. Higgins, J.A. Kain, J. Möller, L. Forchhammer, L. Loft, S. Lorenzo, Y. Collins, A.R. van Schooten, F.J. Laffon, B. Valdiglesias, V. Cooke, M. Mistry, V. Karbaschi, M. Phillips, D.H. Sozeri, O. Routledge, M.N. Nelson-Smith, K. Riso, P. Porrini, M. López de Cerain, A. Azqueta, A. Matullo, G. Allione, A. Møller, P. |
author2_role |
author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Repositório Científico do Instituto Nacional de Saúde |
dc.contributor.author.fl_str_mv |
Godschalk, R.W. Ersson, C. Stępnik, M. Ferlińska, M. Palus, J. Teixeira, João Paulo Costa, S. Jones, G.D. Higgins, J.A. Kain, J. Möller, L. Forchhammer, L. Loft, S. Lorenzo, Y. Collins, A.R. van Schooten, F.J. Laffon, B. Valdiglesias, V. Cooke, M. Mistry, V. Karbaschi, M. Phillips, D.H. Sozeri, O. Routledge, M.N. Nelson-Smith, K. Riso, P. Porrini, M. López de Cerain, A. Azqueta, A. Matullo, G. Allione, A. Møller, P. |
dc.subject.por.fl_str_mv |
DNA Damage Comet Assay Environmental Cancer Risk Ar e Saúde Ocupacional Genotoxicidade Ambiental |
topic |
DNA Damage Comet Assay Environmental Cancer Risk Ar e Saúde Ocupacional Genotoxicidade Ambiental |
description |
This study investigated the levels of DNA strand breaks and formamidopyrimidine DNA glycosylase (FPG) sensitive sites, as assessed by the comet assay, in peripheral blood mononuclear cells (PBMC) from healthy women from five different countries in Europe. The laboratory in each country (referred to as 'centre') collected and cryopreserved PBMC samples from three donors, using a standardised cell isolation protocol. The samples were analysed in 13 different laboratories for DNA damage, which is measured by the comet assay. The study aim was to assess variation in DNA damage in PBMC samples that were collected in the same way and processed using the same blood isolation procedure. The inter-laboratory variation was the prominent contributor to the overall variation. The inter-laboratory coefficient of variation decreased for both DNA strand breaks (from 68 to 26%) and FPG sensitive sites (from 57 to 12%) by standardisation of the primary comet assay endpoint with calibration curve samples. The level of DNA strand breaks in the samples from two of the centres (0.56-0.61 lesions/10(6) bp) was significantly higher compared with the other three centres (0.41-0.45 lesions/10(6) bp). In contrast, there was no difference between the levels of FPG sensitive sites in PBMC samples from healthy donors in the different centres (0.41-0.52 lesion/10(6) bp). |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014-07 2014-07-01T00:00:00Z 2015-02-03T13:10:07Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.18/2752 |
url |
http://hdl.handle.net/10400.18/2752 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Mutagenesis. 2014 Jul;29(4):241-9. doi: 10.1093/mutage/geu012. Epub 2014 Apr 15. 0267-8357 10.1093/mutage/geu012 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/embargoedAccess |
eu_rights_str_mv |
embargoedAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Oxford University Press/UK Environmental Mutagen Society |
publisher.none.fl_str_mv |
Oxford University Press/UK Environmental Mutagen Society |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
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1799132112433971200 |