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

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

Autoren

  • 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

Organisationseinheiten

Externe Organisationen

  • Technische Universität Dortmund
  • Merck & Co., Inc.
  • BASF Innovation Center Ghent
  • Bundesinstitut für Arzneimittel und Medizinprodukte (BfArM)
  • Bayer AG
  • Boehringer Ingelheim
  • Fraunhofer-Institut für Toxikologie und Experimentelle Medizin (ITEM)
  • Institute for Competence Contract Research – Roßdorf (ICCR-Roßdorf) GmbH
  • Procter and Gamble
Forschungs-netzwerk anzeigen

Details

OriginalspracheEnglisch
Aufsatznummer105583
FachzeitschriftRegulatory Toxicology and Pharmacology
Jahrgang148
Frühes Online-Datum22 Feb. 2024
PublikationsstatusVeröffentlicht - März 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.

ASJC Scopus Sachgebiete

  • Pharmakologie, Toxikologie und Pharmazie (insg.)
  • Toxikologie

Zitieren

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, Jahrgang 148, 105583, 03.2024.

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-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, Jg. 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, Artikel 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 Mär;148:105583. Epub 2024 Feb 22. doi: 10.1016/j.yrtph.2024.105583
Download
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title = "In vivo alkaline comet assay: Statistical considerations on historical negative and positive control data",
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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note = "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).",
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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.

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KW - Genotoxicity

KW - Historical control data

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KW - OECD test guideline 489

KW - Rat

KW - Summarizing strategies

KW - Variance components analysis

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