Comparative analysis of baseline 8-oxo-7,8-dihydroguanine in mammalian cell DNA, by different methods in different laboratories: an approach to consensus | Zendy

Andrew Collins | Zendy; C. Gedik | Zendy; Netherlands Vaughan | Zendy; S. Wood | Zendy; J. Dubois | Zendy; Pierre Duez | Zendy; G. Dehon | Zendy; JeanFrançois Rees | Zendy; Steffen Loft | Zendy; P. Moller | Zendy; H. Poulsen | Zendy; B. Riis | Zendy; Allan Weimann | Zendy; Jean Cadet | Zendy; Thierry Douki | Zendy; JeanLuc Ravanat | Zendy; Sylvie Sauvaigo | Zendy; Henri Faure | Zendy; Isabelle Morel | Zendy; Bénédicte Morin | Zendy; Bernd Epe | Zendy; N. Phoa | Zendy; A. Hartwig | Zendy; Alexander Pelzer | Zendy; Piero Dolara | Zendy; Claudia Casalini | Zendy; Lisa Giovannelli | Zendy; M. Lodovici | Zendy; Ryszard Ólinski | Zendy; K. Bialkowski | Zendy; Marek Foksiński | Zendy; Daniel Gackowski | Zendy; Zdeňka Ďuračková | Zendy; L. Hlincikova | Zendy; P. Korytar | Zendy; M. Sivonova | Zendy; Mária Dušinská | Zendy; C. Mislanova | Zendy; José Viña | Zendy; Ana Lloret | Zendy; L. Moller | Zendy; Tim Hofer | Zendy; J. Nygren | Zendy; E. Gremaud | Zendy; Karl E. Herbert | Zendy; D. Chauhan | Zendy; F. Kelly | Zendy; C. Dunster | Zendy; John Lunec | Zendy; Marcus S. Cooke | Zendy; Mark D. Evans | Zendy; Parul Patel | Zendy; I. Podmore | Zendy; A. White | Zendy; C. Wild | Zendy; Laura J. Hardie | Zendy; J. Olliver | Zendy; E. Smith | Zendy

Open Access

Comparative analysis of baseline 8-oxo-7,8-dihydroguanine in mammalian cell DNA, by different methods in different laboratories: an approach to consensus

Author(s) -

Andrew Collins,

C. Gedik,

Netherlands Vaughan,

S. Wood,

J. Dubois,

Pierre Duez,

G. Dehon,

JeanFrançois Rees,

Steffen Loft,

P. Moller,

H. Poulsen,

B. Riis,

Allan Weimann,

Jean Cadet,

Thierry Douki,

JeanLuc Ravanat,

Sylvie Sauvaigo,

Henri Faure,

Isabelle Morel,

Bénédicte Morin,

Bernd Epe,

N. Phoa,

A. Hartwig,

Alexander Pelzer,

Piero Dolara,

Claudia Casalini,

Lisa Giovannelli,

M. Lodovici,

Ryszard Ólinski,

K. Bialkowski,

Marek Foksiński,

Daniel Gackowski,

Zdeňka Ďuračková,

L. Hlincikova,

P. Korytar,

M. Sivonova,

Mária Dušinská,

C. Mislanova,

José Viña,

Ana Lloret,

L. Moller,

Tim Hofer,

J. Nygren,

E. Gremaud,

Karl E. Herbert,

D. Chauhan,

F. Kelly,

C. Dunster,

John Lunec,

Marcus S. Cooke,

Mark D. Evans,

Parul Patel,

I. Podmore,

A. White,

C. Wild,

Laura J. Hardie,

J. Olliver,

E. Smith

Publication year - 2002

Publication title -

carcinogenesis

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.688

H-Index - 204

eISSN - 1460-2180

pISSN - 0143-3334

DOI - 10.1093/carcin/23.12.2129

Subject(s) - hela , dna , comet assay , dna damage , dna glycosylase , oxidative phosphorylation , microbiology and biotechnology , biochemistry , genotoxicity , dna repair , chemistry , biology , cell , toxicity , organic chemistry

The European Standards Committee on Oxidative DNA Damage (ESCODD) was set up to resolve the problems associated with the measurement of background levels of oxidative DNA damage (in particular 8-oxo-7,8-dihydroguanine, or 8-oxoGua) in human cells. A tendency for DNA oxidation to occur during sample preparation prior to chromatography has been recognized as the source of a very substantial artefact. To assess the success of attempts to eliminate the artefact, ESCODD has distributed to its members standard samples of pig liver and HeLa cells for analysis. Estimates of 8-oxoGua in pig liver, using chromatographic techniques, ranged from 2.23 to 441 per 10(6) guanines, with a median of 10.47 per 10(6) guanines. Chromatographic analysis of HeLa cell DNA gave a range of 1.84 to 214 per 10(6) guanines with a median of 5.23 per 10(6) guanines. HeLa cell DNA was also analysed by an enzymic approach, in which whole cell DNA was treated with formamidopyrimidine DNA glycosylase, which nicks DNA at sites of 8-oxoGua, and the breaks measured with the comet assay, alkaline unwinding or alkaline elution. Values with these methods ranged from 0.06 to 4.988-oxoGua per 10(6) guanines, with a median of 0.79 per 10(6) guanines. Although there are clearly still serious discrepancies between methods and laboratories, the lowest estimates by chromatography (arguably those in which the artefact was best controlled) are only 2.5 times higher than the median value obtained with the enzymic approach.

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