Details
| Original language | English |
|---|---|
| Article number | 105583 |
| Journal | Regulatory Toxicology and Pharmacology |
| Volume | 148 |
| Early online date | 22 Feb 2024 |
| Publication status | Published - Mar 2024 |
Abstract
The alkaline comet assay is frequently used as in vivo follow-up test within different regulatory environments to characterize the DNA-damaging potential of different test items. The corresponding OECD Test guideline 489 highlights the importance of statistical analyses and historical control data (HCD) but does not provide detailed procedures. Therefore, the working group “Statistics” of the German-speaking Society for Environmental Mutation Research (GUM) collected HCD from five laboratories and >200 comet assay studies and performed several statistical analyses. Key results included that (I) observed large inter-laboratory effects argue against the use of absolute quality thresholds, (II) > 50% zero values on a slide are considered problematic, due to their influence on slide or animal summary statistics, (III) the type of summarizing measure for single-cell data (e.g., median, arithmetic and geometric mean) may lead to extreme differences in resulting animal tail intensities and study outcome in the HCD. These summarizing values increase the reliability of analysis results by better meeting statistical model assumptions, but at the cost of information loss. Furthermore, the relation between negative and positive control groups in the data set was always satisfactorily (or sufficiently) based on ratio, difference and quantile analyses.
Keywords
- Descriptive statistics, DNA damage, Genotoxicity, Historical control data, In vivo mammalian alkaline comet assay, OECD test guideline 489, Rat, Summarizing strategies, Variance components analysis
ASJC Scopus subject areas
- Pharmacology, Toxicology and Pharmaceutics(all)
- Toxicology
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In: Regulatory Toxicology and Pharmacology, Vol. 148, 105583, 03.2024.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - In vivo alkaline comet assay
T2 - Statistical considerations on historical negative and positive control data
AU - Tug, Timur
AU - Duda, Julia C.
AU - Menssen, Max
AU - Bruce, Shannon Wilson
AU - Bringezu, Frank
AU - Dammann, Martina
AU - Frötschl, Roland
AU - Harm, Volker
AU - Ickstadt, Katja
AU - Igl, Bernd-Wolfgang
AU - Jarzombek, Marco
AU - Kellner, Rupert
AU - Lott, Jasmin
AU - Pfuhler, Stefan
AU - Plappert-Helbig, Ulla
AU - Rahnenführer, Jörg
AU - Schulz, Markus
AU - Vaas, Lea
AU - Vasquez, Marie
AU - Ziegler, Verena
AU - Ziemann, Christina
N1 - The German Society for Environmental Mutation Research (GUM e.V.) supported the work by making a contribution to the open access fees. This work was also supported by the Research Training Group “Biostatistical Methods for High-Dimensional Data in Toxicology” (RTG 2624, Project I1 and P2) funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation—Project Number 427806116).
PY - 2024/3
Y1 - 2024/3
N2 - The alkaline comet assay is frequently used as in vivo follow-up test within different regulatory environments to characterize the DNA-damaging potential of different test items. The corresponding OECD Test guideline 489 highlights the importance of statistical analyses and historical control data (HCD) but does not provide detailed procedures. Therefore, the working group “Statistics” of the German-speaking Society for Environmental Mutation Research (GUM) collected HCD from five laboratories and >200 comet assay studies and performed several statistical analyses. Key results included that (I) observed large inter-laboratory effects argue against the use of absolute quality thresholds, (II) > 50% zero values on a slide are considered problematic, due to their influence on slide or animal summary statistics, (III) the type of summarizing measure for single-cell data (e.g., median, arithmetic and geometric mean) may lead to extreme differences in resulting animal tail intensities and study outcome in the HCD. These summarizing values increase the reliability of analysis results by better meeting statistical model assumptions, but at the cost of information loss. Furthermore, the relation between negative and positive control groups in the data set was always satisfactorily (or sufficiently) based on ratio, difference and quantile analyses.
AB - The alkaline comet assay is frequently used as in vivo follow-up test within different regulatory environments to characterize the DNA-damaging potential of different test items. The corresponding OECD Test guideline 489 highlights the importance of statistical analyses and historical control data (HCD) but does not provide detailed procedures. Therefore, the working group “Statistics” of the German-speaking Society for Environmental Mutation Research (GUM) collected HCD from five laboratories and >200 comet assay studies and performed several statistical analyses. Key results included that (I) observed large inter-laboratory effects argue against the use of absolute quality thresholds, (II) > 50% zero values on a slide are considered problematic, due to their influence on slide or animal summary statistics, (III) the type of summarizing measure for single-cell data (e.g., median, arithmetic and geometric mean) may lead to extreme differences in resulting animal tail intensities and study outcome in the HCD. These summarizing values increase the reliability of analysis results by better meeting statistical model assumptions, but at the cost of information loss. Furthermore, the relation between negative and positive control groups in the data set was always satisfactorily (or sufficiently) based on ratio, difference and quantile analyses.
KW - Descriptive statistics
KW - DNA damage
KW - Genotoxicity
KW - Historical control data
KW - In vivo mammalian alkaline comet assay
KW - OECD test guideline 489
KW - Rat
KW - Summarizing strategies
KW - Variance components analysis
UR - http://www.scopus.com/inward/record.url?scp=85187527277&partnerID=8YFLogxK
U2 - 10.1016/j.yrtph.2024.105583
DO - 10.1016/j.yrtph.2024.105583
M3 - Article
VL - 148
JO - Regulatory Toxicology and Pharmacology
JF - Regulatory Toxicology and Pharmacology
SN - 0273-2300
M1 - 105583
ER -