Permissible level of toxaphene residues in fish from the German market based on in vivo and in vitro effects to tumor promotion

Toxaphene is a chlorinated pesticide consisting of more than 200 congeners that are mainly chlorobornanes and chlorocamphenes. As the congeners exhibit different stability properties in the environment, only between 20 and 30 compounds can be observed in, e.g., fish, which are represented by technical toxaphene as a mixture. In human body, the congeners Parlar #26, #40, #41, #44, #50, and #62 are detected frequently. Three of them, #26, #50, and #62, pose a potential risk to human health due to their persistent characteristic. By using experimental results of a European Union study (MATT, 2000. Investigation into the Monitoring, Analysis and Toxicity of Toxaphene in Marine Foodstuffs, European Union, Brussels, Final report, FAIR CT PL.96.3131. Investigation into the Monitoring, Analysis and Toxicity of Toxaphene in Marine Foodstuffs), a reference dose related to tumor promotion was calculated for these representative persistent toxaphene congeners. In Germany, the sum of the congeners #26, #50, and #62 is defined as the official standard for toxaphene residues in food. In this work, different fish samples obtained from German markets were studied regarding their contamination with toxaphene congeners, presented either in sum, or as single constitutes. The obtained data were used to define the acceptable total concentration of the sum of Parlar #26, #50, and #62 with regard to prevention of tumor promotion in human. The results showed that the currently existing permissible level of the sum of these congeners (0.1 mg/kg) is higher than the acceptable concentration in fish samples determined by this work and calculated at ca. 0.090 mg/kg. It is therefore recommended to improve the permissible level of toxaphene in German food samples.     

Gene expression in two hepatic cell lines, cultured primary hepatocytes, and liver slices compared to the in vivo liver gene expression in rats: possible implications for toxicogenomics use of in vitro systems.

Microarray technology allows the simultaneous analysis of mRNA expression levels of thousands of genes. In the field of toxicogenomics, this technology could help to identify potentially unsafe compounds based on the changes in mRNA expression patterns they induce. Rodent in vivo and in vitro systems are currently the experimental models of choice for predictive toxicology, especially in early phases of development. This study characterizes several hepatic in vitro systems based on mRNA expression profiles, comparing them to gene expression in liver tissue. The in vitro systems investigated comprise two rat liver cell lines (BRL3A and NRL clone 9), primary hepatocytes in conventional monolayer or in sandwich culture, and liver slices. The results demonstrate that liver slices exhibit the strongest similarity to liver tissue regarding mRNA expression, whereas the two cell lines are quite different from the whole liver. We were able to identify genes with strong changes in expression levels in all or at least one of the in vitro systems relative to whole liver. In particular, for some cytochrome P450s the differences observed on the mRNA expression level were paralleled by protein expression and enzymatic activity. In addition, the effect of time in culture was assessed. We were able to show a profound effect of the duration of culture. Expression patterns change most rapidly soon after cell isolation and culture initiation and stabilize with time in culture. The findings are discussed with respect to the usefulness of the various hepatic in vitro systems for microarray-based toxicological testing of compounds.