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Predictive capacity of a non-radioisotopic local lymph node assay using flow cytometry, LLNA: BrdU-FCM: Comparison of a cutoff approach and inferential statistics
- Predictive capacity of a non-radioisotopic local lymph node assay using flow cytometry, LLNA: BrdU-FCM: Comparison of a cutoff approach and inferential statistics
- Kim, Da-eun; Yang, Hyeri; Jang, Won-Hee; Jung, Kyoung-Mi; Park, Miyoung; Choi, Jin Kyu; Jung, Mi-Sook; Jeon, Eun-Young; Heo, Yong; Yeo, Kyung-Wook; Jo, Ji-Hoon; Park, Jung Eun; Sohn, Soo Jung; Kim, Tae Sung; Ahn, Il Young; Jeong, Tae-Cheon; Lim, Kyung-Min; Bae, SeungJin
- Ewha Authors
- 배승진; 임경민
- SCOPUS Author ID
- 배승진; 임경민
- Issue Date
- Journal Title
- JOURNAL OF PHARMACOLOGICAL AND TOXICOLOGICAL METHODS
- 1056-8719; 1873-488X
- vol. 78, pp. 76 - 84
- Local lymph node assay; LLNA: BrdU-FCM; Prediction model; Skin sensitization; Predictive capacity; Descriptive and inferential statistics
- ELSEVIER SCIENCE INC
- SCIE; SCOPUS
- In order for a novel test method to be applied for regulatory purposes, its reliability and relevance, i.e., reproducibility and predictive capacity, must be demonstrated. Here, we examine the predictive capacity of a novel non-radioisotopic local lymph node assay, LLNA: BrdU-FCM (5-bromo-2'-deoxyuridine-flow cytometry), with a cutoff approach and inferential statistics as a prediction model. 22 reference substances in OECD TG429 were tested with a concurrent positive control, hexylcinnamaldehyde 25%(PC), and the stimulation index (SI) representing the fold increase in lymph node cells over the vehicle control was obtained. The optimal cutoff SI (2.7 <= cutoff <3.5), with respect to predictive capacity, was obtained by a receiver operating characteristic curve, which produced 90.9% accuracy for the 22 substances. To address the inter-test variability in responsiveness, SI values standardized with PC were employed to obtain the optimal percentage cutoff (42.6 <= cutoff <57.3% of PC), which produced 86.4% accuracy. A test substance may be diagnosed as a sensitizer if a statistically significant increase in SI is elicited. The parametric one-sided t-test and non-parametric Wilcoxon rank-sum test produced 77.3% accuracy. Similarly, a test substance could be defined as a sensitizer if the SI means of the vehicle control, and of the low, middle, and high concentrations were statistically significantly different, which was tested using ANOVA or Kruskal-Wallis, with post hoc analysis, Dunnett, or DSCF (Dwass-Steel-Critchlow-Fligner), respectively, depending on the equal variance test, producing 81.8% accuracy. The absolute SI-based cutoff approach produced the best predictive capacity, however the discordant decisions between prediction models need to be examined further. (C) 2015 Elsevier Inc. All rights reserved.
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