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Long-term cryopreservation of potassium bromate positive assay controls for measurement of oxidatively damaged DNA by the Fpg-modified comet assay: results from the hCOMET ring trial

2023-10-14Journal article Restricted access
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dc.contributor.authorMøller, Peter
dc.contributor.authorAzqueta, Amaya
dc.contributor.authorRodriguez-Garraus, Adriana
dc.contributor.authorBakuradze, Tamara
dc.contributor.authorRichling, Elke
dc.contributor.authorBankoglu, Ezgi Eyluel
dc.contributor.authorStopper, Helga
dc.contributor.authorClaudino Bastos, Victoria
dc.contributor.authorLangie, Sabine A.S.
dc.contributor.authorJensen, Annie
dc.contributor.authorRistori, Sara
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.authorPaulo Teixeira, João
dc.contributor.authorMarino, Mirko
dc.contributor.authorDel Bo’, Cristian
dc.contributor.authorRiso, Patrizia
dc.contributor.authorZheng, Congying
dc.contributor.authorShaposhnikov, Sergey
dc.contributor.authorCollins, Andrew
dc.date.accessioned2024-02-08T15:01:49Z
dc.date.available2024-02-08T15:01:49Z
dc.date.issued2023-10-14
dc.description.abstractThe formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay is widely used for the measurement of oxidatively generated damage to DNA. However, there has not been a recommended long-term positive control for this version of the comet assay. We have investigated potassium bromate as a positive control for the Fpg-modified comet assay because it generates many Fpg-sensitive sites with a little concurrent generation of DNA strand breaks. Eight laboratories used the same procedure for the treatment of monocytic THP-1 cells with potassium bromate (0, 0.5, 1.5, and 4.5 mM) and subsequent cryopreservation in a freezing medium consisting of 50% foetal bovine serum, 40% RPMI-1640 medium, and 10% dimethyl sulphoxide. The samples were analysed by the Fpg-modified comet assay three times over a 3-year period. All laboratories obtained a positive concentration-response relationship in cryopreserved samples (linear regression coefficients ranging from 0.79 to 0.99). However, there was a wide difference in the levels of Fpg-sensitive sites between the laboratory with the lowest (4.2% Tail DNA) and highest (74% Tail DNA) values in THP-1 cells after exposure to 4.5 mM KBrO3. In an attempt to assess sources of inter-laboratory variation in Fpg-sensitive sites, comet images from one experiment in each laboratory were forwarded to a central laboratory for visual scoring. There was high consistency between measurements of %Tail DNA values in each laboratory and the visual score of the same comets done in the central laboratory (r = 0.98, P < 0.001, linear regression). In conclusion, the results show that potassium bromate is a suitable positive comet assay control.pt_PT
dc.description.sponsorshipCertain authors have acknowledged funding as follows: statutory funding for INTC and IRH (Maria Wojewódzka and Marcin Kruszewski); Spanish Ministry of Science and Innovation MCIN/AEI/10.13039/501100011033 (grants PID2020-113788RB-I00 and PID2020-114908 GA-I00), and Xunta de Galicia (grant ED431B 2022/16) (Vanessa Valdiglesias and Blanca Laffon); Università degli Studi di Firenze fund RICATEN21 and RICATEN22 (Lisa Giovannelli).pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationMutagenesis. 2023 Oct 14;38(5):264-272. doi: 10.1093/mutage/gead020pt_PT
dc.identifier.doi10.1093/mutage/gead020pt_PT
dc.identifier.issn0267-8357
dc.identifier.urihttp://hdl.handle.net/10400.18/9055
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherOxford University Press/ UK Environmental Mutagen Society.pt_PT
dc.relation.publisherversionhttps://academic.oup.com/mutage/article-abstract/38/5/264/7207512?redirectedFrom=fulltextpt_PT
dc.rights.urihttp://creativecommons.org/licenses/by-nc/4.0/pt_PT
dc.subjectFormamidopyrimidine DNA Glycosylaspt_PT
dc.subjectComet Assaypt_PT
dc.subjectDamage to DNApt_PT
dc.subjectPotassiumpt_PT
dc.subjectCryopreservationpt_PT
dc.subjectDimethyl Sulfoxide DNApt_PT
dc.subjectLaboratorypt_PT
dc.subjectMonocytespt_PT
dc.subjectLinear Regressionpt_PT
dc.subjectPositive Controlpt_PT
dc.subjectthp-1 cellspt_PT
dc.titleLong-term cryopreservation of potassium bromate positive assay controls for measurement of oxidatively damaged DNA by the Fpg-modified comet assay: results from the hCOMET ring trialpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage272pt_PT
oaire.citation.issue5pt_PT
oaire.citation.startPage264pt_PT
oaire.citation.titleMutagenesispt_PT
oaire.citation.volume38pt_PT
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT

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