Potassium bromate as positive assay control for the Fpg-modified comet assay

Detalhes bibliográficos
Autor(a) principal: Møller, P
Data de Publicação: 2020
Outros Autores: Muruzabal, D, Bakuradze, T, Richling, E, Bankoglu, EE, Stopper, H, Langie, SAS, Azqueta, A, Jensen, A, Scavone, F, Giovannelli, L, Wojewódzka, M, Kruszewski, M, Valdiglesias, V, Laffon, B, Costa, C, Costa, S, Teixeira, JP, Marino, M, Del, Bo', C, Riso, P, Shaposhnikov, S, Collins, A
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: https://hdl.handle.net/10216/143221
Resumo: The comet assay is a popular assay in biomonitoring studies. DNA strand breaks (or unspecific DNA lesions) are measured using the standard comet assay. Oxidative stress-generated DNA lesions can be measured by employing DNA repair enzymes to recognise oxidatively damaged DNA. Unfortunately, there has been a tendency to fail to report results from assay controls (or maybe even not to employ assay controls). We believe this might have been due to uncertainty as to what really constitutes a positive control. It should go without saying that a biomonitoring study cannot have a positive control group as it is unethical to expose healthy humans to DNA damaging (and thus potentially carcinogenic) agents. However, it is possible to include assay controls in the analysis (here meant as a cryopreserved sample of cells i.e. included in each experiment as a reference sample). In the present report we tested potassium bromate (KBrO3) as a positive comet assay control for the formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay. Ten laboratories used the same procedure for treatment of monocytic THP-1 cells with KBrO3 (0.5, 1.5 and 4.5 mM for 1 h at 37°C) and subsequent cryopreservation. Results from one laboratory were excluded in the statistical analysis because of technical issues in the Fpg-modified comet assay. All other laboratories found a concentration–response relationship in cryopreserved samples (regression coefficients from 0.80 to 0.98), although with different slopes ranging from 1.25 to 11.9 Fpg-sensitive sites (%DNA in tail) per 1 mM KBrO3. Our results demonstrate that KBrO3 is a suitable positive comet assay control.
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spelling Potassium bromate as positive assay control for the Fpg-modified comet assayThe comet assay is a popular assay in biomonitoring studies. DNA strand breaks (or unspecific DNA lesions) are measured using the standard comet assay. Oxidative stress-generated DNA lesions can be measured by employing DNA repair enzymes to recognise oxidatively damaged DNA. Unfortunately, there has been a tendency to fail to report results from assay controls (or maybe even not to employ assay controls). We believe this might have been due to uncertainty as to what really constitutes a positive control. It should go without saying that a biomonitoring study cannot have a positive control group as it is unethical to expose healthy humans to DNA damaging (and thus potentially carcinogenic) agents. However, it is possible to include assay controls in the analysis (here meant as a cryopreserved sample of cells i.e. included in each experiment as a reference sample). In the present report we tested potassium bromate (KBrO3) as a positive comet assay control for the formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay. Ten laboratories used the same procedure for treatment of monocytic THP-1 cells with KBrO3 (0.5, 1.5 and 4.5 mM for 1 h at 37°C) and subsequent cryopreservation. Results from one laboratory were excluded in the statistical analysis because of technical issues in the Fpg-modified comet assay. All other laboratories found a concentration–response relationship in cryopreserved samples (regression coefficients from 0.80 to 0.98), although with different slopes ranging from 1.25 to 11.9 Fpg-sensitive sites (%DNA in tail) per 1 mM KBrO3. Our results demonstrate that KBrO3 is a suitable positive comet assay control.Oxford University Press20202020-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/10216/143221eng0267-83571464-380410.1093/mutage/geaa011Møller, PMuruzabal, DBakuradze, TRichling, EBankoglu, EEStopper, HLangie, SASAzqueta, AJensen, AScavone, FGiovannelli, LWojewódzka, MKruszewski, MValdiglesias, VLaffon, BCosta, CCosta, STeixeira, JPMarino, MDel, Bo', CRiso, PShaposhnikov, SCollins, Ainfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-29T13:09:43Zoai:repositorio-aberto.up.pt:10216/143221Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T23:34:47.090091Repositório 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 Potassium bromate as positive assay control for the Fpg-modified comet assay
title Potassium bromate as positive assay control for the Fpg-modified comet assay
spellingShingle Potassium bromate as positive assay control for the Fpg-modified comet assay
Møller, P
title_short Potassium bromate as positive assay control for the Fpg-modified comet assay
title_full Potassium bromate as positive assay control for the Fpg-modified comet assay
title_fullStr Potassium bromate as positive assay control for the Fpg-modified comet assay
title_full_unstemmed Potassium bromate as positive assay control for the Fpg-modified comet assay
title_sort Potassium bromate as positive assay control for the Fpg-modified comet assay
author Møller, P
author_facet Møller, P
Muruzabal, D
Bakuradze, T
Richling, E
Bankoglu, EE
Stopper, H
Langie, SAS
Azqueta, A
Jensen, A
Scavone, F
Giovannelli, L
Wojewódzka, M
Kruszewski, M
Valdiglesias, V
Laffon, B
Costa, C
Costa, S
Teixeira, JP
Marino, M
Del, Bo', C
Riso, P
Shaposhnikov, S
Collins, A
author_role author
author2 Muruzabal, D
Bakuradze, T
Richling, E
Bankoglu, EE
Stopper, H
Langie, SAS
Azqueta, A
Jensen, A
Scavone, F
Giovannelli, L
Wojewódzka, M
Kruszewski, M
Valdiglesias, V
Laffon, B
Costa, C
Costa, S
Teixeira, JP
Marino, M
Del, Bo', C
Riso, P
Shaposhnikov, S
Collins, A
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Møller, P
Muruzabal, D
Bakuradze, T
Richling, E
Bankoglu, EE
Stopper, H
Langie, SAS
Azqueta, A
Jensen, A
Scavone, F
Giovannelli, L
Wojewódzka, M
Kruszewski, M
Valdiglesias, V
Laffon, B
Costa, C
Costa, S
Teixeira, JP
Marino, M
Del, Bo', C
Riso, P
Shaposhnikov, S
Collins, A
description The comet assay is a popular assay in biomonitoring studies. DNA strand breaks (or unspecific DNA lesions) are measured using the standard comet assay. Oxidative stress-generated DNA lesions can be measured by employing DNA repair enzymes to recognise oxidatively damaged DNA. Unfortunately, there has been a tendency to fail to report results from assay controls (or maybe even not to employ assay controls). We believe this might have been due to uncertainty as to what really constitutes a positive control. It should go without saying that a biomonitoring study cannot have a positive control group as it is unethical to expose healthy humans to DNA damaging (and thus potentially carcinogenic) agents. However, it is possible to include assay controls in the analysis (here meant as a cryopreserved sample of cells i.e. included in each experiment as a reference sample). In the present report we tested potassium bromate (KBrO3) as a positive comet assay control for the formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay. Ten laboratories used the same procedure for treatment of monocytic THP-1 cells with KBrO3 (0.5, 1.5 and 4.5 mM for 1 h at 37°C) and subsequent cryopreservation. Results from one laboratory were excluded in the statistical analysis because of technical issues in the Fpg-modified comet assay. All other laboratories found a concentration–response relationship in cryopreserved samples (regression coefficients from 0.80 to 0.98), although with different slopes ranging from 1.25 to 11.9 Fpg-sensitive sites (%DNA in tail) per 1 mM KBrO3. Our results demonstrate that KBrO3 is a suitable positive comet assay control.
publishDate 2020
dc.date.none.fl_str_mv 2020
2020-01-01T00:00:00Z
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 https://hdl.handle.net/10216/143221
url https://hdl.handle.net/10216/143221
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 0267-8357
1464-3804
10.1093/mutage/geaa011
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dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Oxford University Press
publisher.none.fl_str_mv Oxford University Press
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
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