title: In vivo Comet assay--statistical analysis and power calculations of mice testicular cells.
abstract: The in vivo Comet assay is a sensitive method for evaluating DNA damage. A recurrent concern is how to analyze the data appropriately and efficiently. A popular approach is to summarize the raw data into a summary statistic prior to the statistical analysis. However, consensus on which summary statistic to use has yet to be reached. Another important consideration concerns the assessment of proper sample sizes in the design of Comet assay studies. This study aims to identify a statistic suitably summarizing the % tail DNA of mice testicular samples in Comet assay studies. A second aim is to provide curves for this statistic outlining the number of animals and gels to use. The current study was based on 11 compounds administered via oral gavage in three doses to male mice: CAS no. 110-26-9, CAS no. 512-56-1, CAS no. 111873-33-7, CAS no. 79-94-7, CAS no. 115-96-8, CAS no. 598-55-0, CAS no. 636-97-5, CAS no. 85-28-9, CAS no. 13674-87-8, CAS no. 43100-38-5 and CAS no. 60965-26-6. Testicular cells were examined using the alkaline version of the Comet assay and the DNA damage was quantified as % tail DNA using a fully automatic scoring system. From the raw data 23 summary statistics were examined. A linear mixed-effects model was fitted to the summarized data and the estimated variance components were used to generate power curves as a function of sample size. The statistic that most appropriately summarized the within-sample distributions was the median of the log-transformed data, as it most consistently conformed to the assumptions of the statistical model. Power curves for 1.5-, 2-, and 2.5-fold changes of the highest dose group compared to the control group when 50 and 100 cells were scored per gel are provided to aid in the design of future Comet assay studies on testicular cells.
authors.affiliation: Technical University of Denmark, Department of Applied Mathematics and Computer Science, Matematiktorvet, DK-2800 Kgs. Lyngby, Denmark. Electronic address: mkha@dtu.dk.
authors.forename: Merete Kjær
authors.initials: MK
authors.lastname: Hansen
authors.affiliation: Technical University of Denmark, National Food Institute, Mørkhøj Bygade 19, DK-2860 Søborg, Denmark.
authors.forename: Anoop Kumar
authors.initials: AK
authors.lastname: Sharma
authors.affiliation: Technical University of Denmark, National Food Institute, Mørkhøj Bygade 19, DK-2860 Søborg, Denmark.
authors.forename: Marianne
authors.initials: M
authors.lastname: Dybdahl
authors.affiliation: Technical University of Denmark, National Food Institute, Mørkhøj Bygade 19, DK-2860 Søborg, Denmark.
authors.forename: Julie
authors.initials: J
authors.lastname: Boberg
authors.affiliation: Technical University of Denmark, Department of Applied Mathematics and Computer Science, Matematiktorvet, DK-2800 Kgs. Lyngby, Denmark; Luleå University of Technology, Department of Business Administration, Technology and Social Sciences, Luleå, Sweden.
authors.forename: Murat
authors.initials: M
authors.lastname: Kulahci
mesh_terms: D000818:Animals; D020552:Comet Assay / Q000379:methods* / Q000706:statistics & numerical data; D004249:DNA Damage; D003627:Data Interpretation, Statistical; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D015233:Models, Statistical; D009153:Mutagens / Q000633:toxicity; D013737:Testis / Q000473:pathology*
chemical_list: D009153:Mutagens
keywords: DNA damage; Genotoxicity; Germ cells; Power; Single-cell gel electrophoresis; Statistical analysis