[Cys-Flanked Cationic Peptides For Cell Delivery of the Herpes Simplex Virus Thymidine Kinase Gene for Suicide Gene Therapy of Uterine Leiomyoma].

Uterine leiomyoma (UL) is the most common benign tumor in women of reproductive age. Gene therapy using suicidal genes appears to be a promising approach for UL treatment. One of key factors for success of gene therapy is the right choice of genetic construct carrier. A promising group of non-viral carriers for cell delivery of expression vectors is cationic Cys-flanked peptides which form tight complexes with DNA due to electrostatic interactions and the presence of interpeptide disulfide bonds. The paper reports a comparative study of the physico-chemical, toxic, and transfectional properties of the DNA-peptide complexes obtained by matrix polymerization or oxidative polycondensation of Cys-flanked peptides using the chain growth terminator 2-amino ethanethiol. We have demonstrated the therapeutic effect of the delivery of the pPTK-1 plasmid carrying the herpes simplex virus type 1 (HSV-1) thymidine kinase gene into PANC-1, and HEK-293T cell culture as well as into primary UL cells. It has been shown that the carriers obtained by oxidative polycondensation transform primary UL cells more efficiently than those produced by matrix polymerization. Treatment with ganciclovir resulted in the death of up to 40% of UL cells transfected with the pPTK-1 plasmid. The perspectives of use of the polyR6 carrier produced by oxidative polycondensation as a tool for the development of modular peptide carriers for the purposes of UL gene therapy were discussed.

[The application of Fluo-3 AM in measurement of level of cytoplasmic calcium in thrombocytes by flow cytofluorometry].

The study was carried out to evaluate possibility of applying technique of thrombin-induced increasing of concentration of Ca in cytoplasm of Fluo-3-colored thrombocytes as an experimental model of studying mechanism of action of anti-thrombocytes medications in vitro. The effect of anti-thrombocyte substances on thrombin-induced increasing of the level of cytoplasmic Ca in thrombocytes was analyzed on example of acetylsalicylic acid. The measurement of concentration of cytoplasmic Ca was implemented using flow cytometry technique with fluorescent probe Fluo-3 AM. It is established that in the given test acetylsalicylic acid inhibits thrombin-induced increasing of cytoplasmic Ca at 0.125-5.0 mk/mol concentrations. This occurrence testifies that in the mechanism of effect of acetylsalicylic acid the suppression of thromboxane path ceases to be a leading one. The proposed methodical approach permits evaluating anti-thrombocite effect of substances according their impact to the level of cytoplasmic Ca in thrombocytes in vitro. However, this approach has a number of limitations preventing wide-spread application of the given technique.

Cytokines and placental macrophages in regulation of birth activity.

The article reviews present notions on functional activity of cytokines of the fetoplacental complex. Particular emphasis is placed on the role of these molecules in the regulation of gestation processes and in pregnancy incompetence. The mechanism of the involvement of placental macrophages and their products in gestation and delivery is discussed.