Statistical tools for analysing the data obtained from repeated dose toxicity studies with rodents: A comparison of the statistical tools used in Japan with that of used in other countries.

In the present study, an attempt was made to compare the statistical tools used for analysing the data of repeated dose toxicity studies with rodents conducted in 45 countries, with that of Japan. The study revealed that there was no congruence among the countries in the use of statistical tools for analysing the data obtained from the above studies. For example, to analyse the data obtained from repeated dose toxicity studies with rodents, Scheffé’s multiple range and Dunnett type (joint type Dunnett) tests are commonly used in Japan, but in other countries use of these statistical tools is not so common. However, statistical techniques used for testing the above data for homogeneity of variance and inter-group comparisons do not differ much between Japan and other countries. In Japan, the data are generally not tested for normality and the same is true with the most of the countries investigated. In the present investigation, out of 127 studies examined, data of only 6 studies were analysed for both homogeneity of variance and normal distribution. For examining homogeneity of variance, we propose Levene’s test, since the commonly used Bartlett’s test may show heterogeneity in variance in all the groups, if a slight heterogeneity in variance is seen any one of the groups. We suggest the data may be examined for both homogeneity of variance and normal distribution. For the data of the groups that do not show heterogeneity of variance, to find the significant difference among the groups, we recommend Dunnett’s test, and for those show heterogeneity of variance, we recommend Steel’s test.

Do we need to examine the quantitative data obtained from toxicity studies for both normality and homogeneity of variance?

Most of the statistical techniques used to evaluate the data obtained from toxicity studies are based on the assumption that the data show a normal distribution and homogeneity of variance. Literature review on toxicity studies on laboratory animals reveals that in most of the cases homogeneity of variance alone is examined for the data obtained from these studies. But the data that show homogeneity of variance need not always show a normal distribution. In fact, most of the data derived from toxicity studies, including hematological and biochemical parameters show a non-normal distribution. On examining normality of data obtained from various toxicity studies using different normality tests, we observed that Shapiro-Wilk test is more appropriate than Kolmogorov-Smimov test, Lilliefors test, the normal probability paper analysis and Chi square test. But there are situations, especially in the long-term toxicity studies, where normality is not shown by one or more than one of the dosage groups. In this situation, we propose that the data maybe analyzed using Dunnett multiple comparison test after excluding the data of the groups that do not show normality However, the biological relevance of the excluded data has to be carefully scrutinized. We also observed that the tendency of the data to show a normal distribution seems to be related to the age of the animals. Present paper describes various tests commonly used to test normality and their power, and also emphasizes the need of subjecting the data obtained from toxicity studies to both normality and homogeneity tests. A flow chart suggesting the statistical techniques that maybe used for both the types of data showing a normal or non-normal distribution is also proposed.

Molecular size as a limiting characteristic for bioconcentration in fish.

The relationships between the bioconcentration factor (BCF) of chemicals in fish and their size, as characterized by molecular weight (MW), effective cross sectional diameter (Deff), and maximum diameter (Dmax) have been investigated using an experimental data set of 737 new and 441 existing chemicals monitored by the Japanese Chemical Substances Control Law (CSCL). Substances with BCF > or = 5000 (very high bioconcentration potential) typically have MW < 550, Deff < 1.1 nm and Dmax < 2.0 nm, respectively and the substances with BCF > or = 1000 (high bioconcentration potential) have MW < 550, Deff < 1.4 nm and Dmax < 2.9 nm, respectively Therefore, the previously suggested threshold values for Deff (0.95 nm) and Dmax (1.5 nm) used for discriminating between bioconcentrative and non-bioconcentrative substances were found to be somewhat small. We found that many substances with BCF > or = 1000 and Dmax > or = 1.5 nm have Deff < 0.95 nm.

Darkened Xenopus tadpoles appeared with neurochemical agents.

Skin darkened tadpoles sometimes appear spontaneously. Darkened was artificially induced in Xenopus larvae by yohimbine or chlorpromazine. These phenomena look like that are seen at pinealectomized or hypothalamus separated Xenopus larva. In this experiment, such a morphological color changed Xenopus larva is suggested by cause of inhibition of alpha2-adrenargic receptor or dopamine receptor from gastrula stages.