A new method of age estimation based on the changes in human non-collagenous proteins from dentin.

The new method exploits the characteristic of staphylococcal protease V8, which specifically splits peptide bonds where l-glutamic or l-aspartic acid participate. Those peptide bonds where d-aspartic acid is present remain unsplit because of the stereospecifity of enzymes. In accordance with expectations, fewer peptide bonds are split by this protease at more advanced ages and larger peptide fragments are thus formed due to the higher content of d-amino acid residues in the proteins of older people. The samples of acid-extracted non-collagenous proteins from dentin were separated using high performance liquid chromatography after enzymatic hydrolysis. A peak with a retention time of 45.3 min was chosen and his enlarging area showed a linear correlation with increasing age. Although the linear correlation with age was proved, the scattering of values decreases the usefulness of the proposed method for age estimation.

Evaluation of toxicity of pesticides and their biodegradation products using human cells.

Juvenoids are biologically active compounds, of relatively low toxicity to humans, that efficiently inhibit the fertility of insects. However, little attention has been paid to the stability and toxicity of products that may be generated by their biodegradation in the ecosystem. This study describes a simple comparison of the toxicity of the active compound and its degradation products generated by aerobic soil microbial isolates. Surprisingly we have found that toxicity of a biologically active carbamate juvenoid N-[2-[4-(2,2-ethylenedioxy-1-cyclohexylmethyl)-phenoxylethyl]carbamate (W328) was comparable with that of 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT). The toxic effect was evaluated using the determination of the ATP/ADP content and viability of HeLa S3 cells exposed to various concentrations of the chemicals tested for various durations. DDT was used as a reference compound. Its toxicity was compared with two juvenile hormone analogs. The original compound, W328, was found to be the most toxic. The major product (W329) generated both by yeast isolates and the mixture of moulds lost its activity on reproduction of the tested insect. Its toxicity towards human cells was also decreased. Another two W328 degradation HPLC fractions exhibited significantly reduced toxicity compared to W328.