[Application of molecular-genetic technologies for the identification of Russian citizens killed in Thailand during tsunami (II): too early to draw the line?].

The Federal State Institution "Russian Centre of Forensic Medical Expertise", Russian Health and Social Development Agency (Roszdrav), carries out forensic medical and molecular-genetic investigations with the purpose of identifying remains of Russian citizens allegedly killed (officially regarded as unaccounted-for) during the recent Thailand's tsunami. The last of the identified Russian subjects was a child from the B---lovs family missed in the disaster area. The child's parents were the last Russians whose fate thus far remained unknown: they were not reported as survivors, nor were their remains found and identified in the course of forensic medical expert investigations. The matter was considered settled. However, this conclusion proved premature. In what follows, we provide arguments that convinced us of the necessity to turn to this issue again and continue the identification studies. As a result of renewed examination, we managed to identify one more of the missed subjects (the child's father). Scientific and technical aspects of the above work that made it possible to reconsider earlier findings and obtain new information are discussed. Special emphasis is laid on the value of molecular-genetic identification of personality as a method of evidence-based forensic medical examination.

[Optimization of methods for DNA extraction from objects of forensic medical examination].

An optimized method for DNA extraction from materials available for forensic medical examination is proposed based on partial combination of two procedures, phenol-chloroform extraction and sorption. Such a combination provided a basis for the development of a protocol making use of the advantages of both procedures and free of drawbacks of either of them. This relatively simple and rapid method uses no reagents having adverse effect on human organism. Also, it excludes the use of substances interfering with polymerase chain reaction (PCR) and thereby reduces the risk of DNA loss. This makes it suitable for the work with objects having very low DNA content. The DNA specimens obtained with the help of the proposed technique are fit for prolonged storage.

[Some practical notes about performing polymerase chain reaction and electrophoretic fractionation of amplificated products].

Practical aspects concerning standardization of molecular-genetic expertise performing with the use of the method of DNA are considered. Examples of difficulties, which can occur at nonobservance of requirements of polymerase chain reaction and electrophoresis performing, are described; practical recommendations of their elimination are given.

[Typing of ABO locus with biological microchip as a new level in solution of problems in forensic-medical biological expertise of material evidences].

There are cases in practice when during expertise of material evidences, discrepancies between results of typing of ABO antigens and molecular-genetic typing of DNA occur. In this work, as a radical approach to objective solution of similar conflict situations, for some contradictory case of expertise, all examinations were performed on the unified methodological base--DNA level. Instead of biological (isoserological) typing of ABO antigen, molecular-genetic typing of ABO locus with biological microchip was performed. In all cases the results, received with the use of biological microchip, do not contradict but completely conform to the results of others molecular-genetic examinations performed in the case. Given results indicate irrationality of further use of traditional methods of isoserological typing of ABO antigen for primary differentiation of biological material. These analyses, if necessary, have to be performed on DNA level with molecular-genetic expertise.

[Activation of RHOA gene transcription in epithelial tumors may be caused by gene amplification and/or demethylation of its promotor region].

RHOA protein, a member of small GTPases family, is implicated in cell morphogenesis, adhesion, and in cell cycle regulation. RHOA gene (3p21.31) exhibits cell transformation activity, and therefore gene is considered as a potential oncogene. The aim of this study was to investigate RHOA transcription and copy number changes in three epithelial tumors (breast, renal cell and epithelial ovarian carcinomas, 45 tumor/normal pairs altogether). EII, HhaI, AciI n Bsh1236I). Hypomethylation of the RHOA promoter region in tumor DNA was observed two times more frequently than increased methylation. Moreover, all (15) cancer cases with hypomethylation of the RHOA gene showed a 2-10 fold increased expression of RHOA. It was concluded that gene copy multiplication and demethylation of the RHOA promoter region can contribute to transcription activation of this gene in epithelial tumors.