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

2020-07Journal article Open access
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dc.contributor.authorMøller, Peter
dc.contributor.authorMuruzabal, Damian
dc.contributor.authorBakuradze, Tamara
dc.contributor.authorRichling, Elke
dc.contributor.authorBankoglu, Ezgi Eyluel
dc.contributor.authorStopper, Helga
dc.contributor.authorLangie, Sabine A.S.
dc.contributor.authorAzqueta, Amaya
dc.contributor.authorJensen, Annie
dc.contributor.authorScavone, Francesca
dc.contributor.authorGiovannelli, Lisa
dc.contributor.authorWojewódzka, Maria
dc.contributor.authorKruszewski, Marcin
dc.contributor.authorValdiglesias, Vanessa
dc.contributor.authorLaffon, Blanca
dc.contributor.authorCosta, Carla
dc.contributor.authorCosta, Solange
dc.contributor.authorTeixeira, João Paulo
dc.contributor.authorMarino, Mirko
dc.contributor.authorDel Bo’, Cristian
dc.contributor.authorRiso, Patrizia
dc.contributor.authorShaposhnikov, Sergey
dc.contributor.authorCollins, Andrew
dc.date.accessioned2021-03-04T11:14:54Z
dc.date.available2021-03-04T11:14:54Z
dc.date.issued2020-07
dc.description.abstractThe 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.pt_PT
dc.description.sponsorshipThe authors thank the hCOMET project (COST Action, CA15132) for support. Carla Costa and Solange Costa were supported by grants FCT-SFRH/BPD/96196/2013 and SFRH/BPD/100948/2014, respectivelypt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationMutagenesis. 2020 Jul;35(4):341-348. Epub 2020 Apr 22. doi: 10.1093/mutage/geaa011pt_PT
dc.identifier.doi10.1093/mutage/geaa011pt_PT
dc.identifier.issn0267-8357
dc.identifier.urihttp://hdl.handle.net/10400.18/7303
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherOxford University Press/ United Kingdom Environmental Mutagen Societypt_PT
dc.relationImpact of transplacental exposure to tobacco smoke in the DNA of newborn. Evaluation of genetic damage and epigenetic changes. Evolution throughout childhood.
dc.relationNew molecular biomarkers for early detection and prevention of frailty in older adults
dc.relation.publisherversionhttps://academic.oup.com/mutage/article/35/4/341/5823763pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectPotassiumpt_PT
dc.subjectBromatept_PT
dc.subjectComet Assaypt_PT
dc.subjectCryopreservationpt_PT
dc.subjectDNApt_PT
dc.subjectLaboratorypt_PT
dc.subjectBiological Monitoringpt_PT
dc.subjectPositive Controlpt_PT
dc.subjectTHP-1 cellspt_PT
dc.subjectGenotoxicidade Ambientalpt_PT
dc.titlePotassium bromate as positive assay control for the Fpg-modified comet assaypt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleImpact of transplacental exposure to tobacco smoke in the DNA of newborn. Evaluation of genetic damage and epigenetic changes. Evolution throughout childhood.
oaire.awardTitleNew molecular biomarkers for early detection and prevention of frailty in older adults
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBPD%2F96196%2F2013/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBPD%2F100948%2F2014/PT
oaire.citation.endPage348pt_PT
oaire.citation.issue4pt_PT
oaire.citation.startPage341pt_PT
oaire.citation.titleMutagenesispt_PT
oaire.citation.volume35pt_PT
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.embargofctAcesso de acordo com página web do editor da revista.pt_PT
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isProjectOfPublicationf4b22fdf-07e9-4ee8-884a-ea80e97bc850
relation.isProjectOfPublication7f2100da-8808-4e66-9c4e-69d3f6112100
relation.isProjectOfPublication.latestForDiscovery7f2100da-8808-4e66-9c4e-69d3f6112100

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