title: Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories.
abstract: 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).
authors.affiliation: Department of Toxicology, Maastricht University, Universiteitssingel 50, Maastricht, Limburg 6200MD, the Netherlands, Department of Biosciences and Nutrition, Karolinska Institutet, Hälsovägen 7-9, Huddinge 14183, Stockholm, Sweden, Toxicology and Carcinogenesis Department, Nofer Institute of Occupational Medicine, 91-348 Łódź, Poland, Environmental Health Department, National Institute of Health, Rua Alexandre Herculano 321, Porto 4000-055, Portugal, Department of Cancer Studies and Molecular Medicine, University of Leicester, Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester LE2 7LX, UK, Department of Public Health, University of Copenhagen, Section of Environmental Health, Øster Farimagsgade 5, Copenhagen K 1014, Denmark, Department of Nutrition, Faculty of Medicine, University of Oslo, PB1046 Blindern, Oslo 0316, Norway, DICOMOSA Group, Department of Psychology, Area of Psychobiology, Edificio de Servicios Centrales de Investigación, Universidade da Coruña, Campus Elviña s/n, 15071-A, Coruña, Spain, Unit of Clinical and Molecular Epidemiology, IRCCS San Raffaele Pisana, Via di Val Cannuta, 247, 00166, Rome, Italy, Oxidative Stress Group, Department Cancer Studies and Molecular Medicine, University of Leicester, Leicester LE2 7LX, UK, Analytical and Environmental Sciences Division, School of Biomedical Sciences, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK, Leeds Institute of Genetics, Health and Therapeutics, University of Leeds, Leeds LS2 9JT, UK, Department of Food Science and Microbiology, Division of Human Nutrition, University of Milan, via Celoria 2, Milan 20133, Italy, Department Food Sciences and Toxicology, University of Navarra, C/ Irunlarrea 1, Pamplona 31008, Spain, Department of Medical Sciences, Via Santena 19, University of Turin, Turin 10126, Italy, Human Genetics Foundation (HuGeF), Via Nizza 52,Turin 10126, Italy.
authors.forename: Roger W L
authors.initials: RW
authors.lastname: Godschalk
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Hälsovägen 7-9, Huddinge 14183, Stockholm, Sweden.
authors.forename: Clara
authors.initials: C
authors.lastname: Ersson
authors.affiliation: Toxicology and Carcinogenesis Department, Nofer Institute of Occupational Medicine, 91-348 Łódź, Poland.
authors.forename: Maciej
authors.initials: M
authors.lastname: Stępnik
authors.affiliation: Toxicology and Carcinogenesis Department, Nofer Institute of Occupational Medicine, 91-348 Łódź, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Ferlińska
authors.affiliation: Toxicology and Carcinogenesis Department, Nofer Institute of Occupational Medicine, 91-348 Łódź, Poland.
authors.forename: Jadwiga
authors.initials: J
authors.lastname: Palus
authors.affiliation: Environmental Health Department, National Institute of Health, Rua Alexandre Herculano 321, Porto 4000-055, Portugal.
authors.forename: João Paulo
authors.initials: JP
authors.lastname: Teixeira
authors.affiliation: Toxicology and Carcinogenesis Department, Nofer Institute of Occupational Medicine, 91-348 Łódź, Poland.
authors.forename: Solange
authors.initials: S
authors.lastname: Costa
authors.affiliation: Department of Cancer Studies and Molecular Medicine, University of Leicester, Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester LE2 7LX, UK.
authors.forename: George D D
authors.initials: GD
authors.lastname: Jones
authors.affiliation: Department of Cancer Studies and Molecular Medicine, University of Leicester, Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester LE2 7LX, UK.
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Higgins
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Hälsovägen 7-9, Huddinge 14183, Stockholm, Sweden.
authors.forename: Johanna
authors.initials: J
authors.lastname: Kain
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Hälsovägen 7-9, Huddinge 14183, Stockholm, Sweden.
authors.forename: Lennart
authors.initials: L
authors.lastname: Möller
authors.affiliation: Department of Public Health, University of Copenhagen, Section of Environmental Health, Øster Farimagsgade 5, Copenhagen K 1014, Denmark.
authors.forename: Lykke
authors.initials: L
authors.lastname: Forchhammer
authors.affiliation: Department of Public Health, University of Copenhagen, Section of Environmental Health, Øster Farimagsgade 5, Copenhagen K 1014, Denmark.
authors.forename: Steffen
authors.initials: S
authors.lastname: Loft
authors.affiliation: Department of Nutrition, Faculty of Medicine, University of Oslo, PB1046 Blindern, Oslo 0316, Norway.
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Lorenzo
authors.affiliation: Department of Nutrition, Faculty of Medicine, University of Oslo, PB1046 Blindern, Oslo 0316, Norway.
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Collins
authors.affiliation: Department of Toxicology, Maastricht University, Universiteitssingel 50, Maastricht, Limburg 6200MD, the Netherlands, Department of Biosciences and Nutrition, Karolinska Institutet, Hälsovägen 7-9, Huddinge 14183, Stockholm, Sweden, Toxicology and Carcinogenesis Department, Nofer Institute of Occupational Medicine, 91-348 Łódź, Poland, Environmental Health Department, National Institute of Health, Rua Alexandre Herculano 321, Porto 4000-055, Portugal, Department of Cancer Studies and Molecular Medicine, University of Leicester, Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester LE2 7LX, UK, Department of Public Health, University of Copenhagen, Section of Environmental Health, Øster Farimagsgade 5, Copenhagen K 1014, Denmark, Department of Nutrition, Faculty of Medicine, University of Oslo, PB1046 Blindern, Oslo 0316, Norway, DICOMOSA Group, Department of Psychology, Area of Psychobiology, Edificio de Servicios Centrales de Investigación, Universidade da Coruña, Campus Elviña s/n, 15071-A, Coruña, Spain, Unit of Clinical and Molecular Epidemiology, IRCCS San Raffaele Pisana, Via di Val Cannuta, 247, 00166, Rome, Italy, Oxidative Stress Group, Department Cancer Studies and Molecular Medicine, University of Leicester, Leicester LE2 7LX, UK, Analytical and Environmental Sciences Division, School of Biomedical Sciences, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK, Leeds Institute of Genetics, Health and Therapeutics, University of Leeds, Leeds LS2 9JT, UK, Department of Food Science and Microbiology, Division of Human Nutrition, University of Milan, via Celoria 2, Milan 20133, Italy, Department Food Sciences and Toxicology, University of Navarra, C/ Irunlarrea 1, Pamplona 31008, Spain, Department of Medical Sciences, Via Santena 19, University of Turin, Turin 10126, Italy, Human Genetics Foundation (HuGeF), Via Nizza 52,Turin 10126, Italy.
authors.forename: Frederik-Jan
authors.initials: FJ
authors.lastname: van Schooten
authors.affiliation: DICOMOSA Group, Department of Psychology, Area of Psychobiology, Edificio de Servicios Centrales de Investigación, Universidade da Coruña, Campus Elviña s/n, 15071-A, Coruña, Spain.
authors.forename: Blanca
authors.initials: B
authors.lastname: Laffon
authors.affiliation: DICOMOSA Group, Department of Psychology, Area of Psychobiology, Edificio de Servicios Centrales de Investigación, Universidade da Coruña, Campus Elviña s/n, 15071-A, Coruña, Spain, Unit of Clinical and Molecular Epidemiology, IRCCS San Raffaele Pisana, Via di Val Cannuta, 247, 00166, Rome, Italy.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Valdiglesias
authors.affiliation: Oxidative Stress Group, Department Cancer Studies and Molecular Medicine, University of Leicester, Leicester LE2 7LX, UK.
authors.forename: Marcus
authors.initials: M
authors.lastname: Cooke
authors.affiliation: Oxidative Stress Group, Department Cancer Studies and Molecular Medicine, University of Leicester, Leicester LE2 7LX, UK.
authors.forename: Vilas
authors.initials: V
authors.lastname: Mistry
authors.affiliation: Oxidative Stress Group, Department Cancer Studies and Molecular Medicine, University of Leicester, Leicester LE2 7LX, UK.
authors.forename: Mahsa
authors.initials: M
authors.lastname: Karbaschi
authors.affiliation: Analytical and Environmental Sciences Division, School of Biomedical Sciences, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: David H
authors.initials: DH
authors.lastname: Phillips
authors.affiliation: Analytical and Environmental Sciences Division, School of Biomedical Sciences, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: Osman
authors.initials: O
authors.lastname: Sozeri
authors.affiliation: Leeds Institute of Genetics, Health and Therapeutics, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: Michael N
authors.initials: MN
authors.lastname: Routledge
authors.affiliation: Leeds Institute of Genetics, Health and Therapeutics, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: Kirsty
authors.initials: K
authors.lastname: Nelson-Smith
authors.affiliation: Department of Food Science and Microbiology, Division of Human Nutrition, University of Milan, via Celoria 2, Milan 20133, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Riso
authors.affiliation: Department of Food Science and Microbiology, Division of Human Nutrition, University of Milan, via Celoria 2, Milan 20133, Italy.
authors.forename: Marisa
authors.initials: M
authors.lastname: Porrini
authors.affiliation: Department Food Sciences and Toxicology, University of Navarra, C/ Irunlarrea 1, Pamplona 31008, Spain.
authors.forename: Adela
authors.initials: A
authors.lastname: López de Cerain
authors.affiliation: Department of Nutrition, Faculty of Medicine, University of Oslo, PB1046 Blindern, Oslo 0316, Norway, Department Food Sciences and Toxicology, University of Navarra, C/ Irunlarrea 1, Pamplona 31008, Spain.
authors.forename: Amaya
authors.initials: A
authors.lastname: Azqueta
authors.affiliation: Department of Medical Sciences, Via Santena 19, University of Turin, Turin 10126, Italy, Human Genetics Foundation (HuGeF), Via Nizza 52,Turin 10126, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Matullo
authors.affiliation: Human Genetics Foundation (HuGeF), Via Nizza 52,Turin 10126, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Allione
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Hälsovägen 7-9, Huddinge 14183, Stockholm, Sweden, pemo@sund.ku.dk.
authors.forename: Peter
authors.initials: P
authors.lastname: Møller
mesh_terms: D000328:Adult; D002138:Calibration; D002469:Cell Separation / Q000379:methods*; D020552:Comet Assay; D053903:DNA Breaks, Double-Stranded; D004249:DNA Damage; D043382:DNA-Formamidopyrimidine Glycosylase / Q000378:metabolism; D005260:Female; D006801:Humans; D007753:Laboratories; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008875:Middle Aged; D009152:Mutagenicity Tests; D012044:Regression Analysis
chemical_list: D043382:DNA-Formamidopyrimidine Glycosylase