In vivo alkaline comet assay: Statistical considerations on historical negative and positive control data

Research output: Contribution to journalArticleResearchpeer review

Authors

  • Timur Tug
  • Julia C. Duda
  • Max Menssen
  • Shannon Wilson Bruce
  • Frank Bringezu
  • Martina Dammann
  • Roland Frötschl
  • Volker Harm
  • Katja Ickstadt
  • Bernd-Wolfgang Igl
  • Marco Jarzombek
  • Rupert Kellner
  • Jasmin Lott
  • Stefan Pfuhler
  • Ulla Plappert-Helbig
  • Jörg Rahnenführer
  • Markus Schulz
  • Lea Vaas
  • Marie Vasquez
  • Verena Ziegler
  • Christina Ziemann

Research Organisations

External Research Organisations

  • TU Dortmund University
  • Merck & Co., Inc.
  • BASF Innovation Center Ghent
  • Federal Institute for Drugs and Medical Devices (BfArM)
  • Bayer AG
  • Boehringer Ingelheim
  • Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM)
  • Institute for Competence Contract Research – Roßdorf (ICCR-Roßdorf) GmbH
  • Procter and Gamble
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Details

Original languageEnglish
Article number105583
JournalRegulatory Toxicology and Pharmacology
Volume148
Early online date22 Feb 2024
Publication statusPublished - 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

Cite this

In vivo alkaline comet assay: Statistical considerations on historical negative and positive control data. / Tug, Timur; Duda, Julia C.; Menssen, Max et al.
In: Regulatory Toxicology and Pharmacology, Vol. 148, 105583, 03.2024.

Research output: Contribution to journalArticleResearchpeer review

Tug, T, Duda, JC, Menssen, M, Bruce, SW, Bringezu, F, Dammann, M, Frötschl, R, Harm, V, Ickstadt, K, Igl, B-W, Jarzombek, M, Kellner, R, Lott, J, Pfuhler, S, Plappert-Helbig, U, Rahnenführer, J, Schulz, M, Vaas, L, Vasquez, M, Ziegler, V & Ziemann, C 2024, 'In vivo alkaline comet assay: Statistical considerations on historical negative and positive control data', Regulatory Toxicology and Pharmacology, vol. 148, 105583. https://doi.org/10.1016/j.yrtph.2024.105583
Tug, T., Duda, J. C., Menssen, M., Bruce, S. W., Bringezu, F., Dammann, M., Frötschl, R., Harm, V., Ickstadt, K., Igl, B.-W., Jarzombek, M., Kellner, R., Lott, J., Pfuhler, S., Plappert-Helbig, U., Rahnenführer, J., Schulz, M., Vaas, L., Vasquez, M., ... Ziemann, C. (2024). In vivo alkaline comet assay: Statistical considerations on historical negative and positive control data. Regulatory Toxicology and Pharmacology, 148, Article 105583. https://doi.org/10.1016/j.yrtph.2024.105583
Tug T, Duda JC, Menssen M, Bruce SW, Bringezu F, Dammann M et al. In vivo alkaline comet assay: Statistical considerations on historical negative and positive control data. Regulatory Toxicology and Pharmacology. 2024 Mar;148:105583. Epub 2024 Feb 22. doi: 10.1016/j.yrtph.2024.105583
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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.",
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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

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