[Implication of sirtuins and kisspeptin in ovarian aging.]

The aim of the work was to study the expression of sirtuins 1 and 6 and kisspeptin in human ovarian tissue taken in different periods of ontogenesis: from the formation of a pool of follicles in the antenatal period to the extinction of ovarian function in postmenopausal women. It was revealed that sirtuins are expressed in human ovary tissue in all age groups. The maximum expression level of SIRT1 in ovarian tissue was observed during intrauterine development and during the perimenopause, SIRT6 during the reproductive period and perimenopause. The participation of SIRT1 in prenatal selection of oocytes in human fetuses was shown, since it was in this group that increased expression of this marker was revealed. The expression level of the kisspeptin marker increases with the formation of the ovaries and the inclusion of reproductive function; the peak of expression is observed in the period before the onset of menopause. The study conducted to identify the expression of these proteins in the human ovary expands the understanding of the regulation of the ovarian follicular reserve and reproductive aging.

[Accumulation of doxorubicin conjugates with dendritic polymer and vector protein in normal and tumor cells in vitro].

Accumulation of doxorubicin (Dox), its conjugates with the second generation dendritic polymer (G2-Dox) and vector pro- tein (recombinant third domain of alpha-fetoprotein - 3D-G2- Dox) in normal and tumor cells was studied in vitro within the framework of the development of selective transport system of anticancer drugs to the target cells. The objects of the study were cells of peripheral blood mononuclear fraction of healthy donors and cells of breast adenocarcinoma lines MCF-7 and MCF-7/MDR1, differing in chemosensitivity. G2-Dox and 3D-G2-Dox accumulated in tumor cells of the both lines better than free Dox (p<0,05). However removal of these drugs out of cells MCF-7 and MCF-7/MDR1 was significantly different: in the latter case all free Dox was excluded from the cells for 24 hours while Dox, accumulated in composition with dendrimers, still remained in the cells. It was important that 3D-G2-Dox (unlike the G2-Dox) accumulated in normal cells worse than free Dox (p<0.01). Thus, the results indicate that the use of 3D-G2-Dox is the most promising because it accumulates in tumor cells better and in normal cells worse than free Dox. Furthermore it can be assumed that the use of 3D-G2-Dox would be especially useful in cases of multi-drug resistance associated with the high expression of P-glycoprotein.

[Increase in the number of cancer stem cells after exposure to low-LET radiation].

Radioresistance of cancer stem cells (CSCs) is regarded as one of the possible causes of cancer recurrence after radiotherapy. Since the regularities and mechanisms of radiation effects on this population of cells have not been sufficiently studied, the aim of this work is to elucidate the changes in the CSC number after γ-irradiation in stable cultures of tumor cells in vitro and tumor tissue in vivo (in the course of radiation therapy of patients with cancers of the upper respiratory tract). CSCs were identified in the cell lines B16, MCF-7, HeLa by the ability to exclude the fluorescent dye Hoechst 33342 (SP method) 48-72 h after irradiation at the doses of 1-20 Gy and in biopsy material by immunophenotype CD44+CD24(-/low) before and 24 h after irradiation at the total dose of 10 Gy. The essential differences in the response of CSCs and other cancer cells were found after exposure to low-LET radiation. The absolute number of CSCs increased after a single exposure at the doses ranging from 1 to 5-10 Gy in different cell cultures, but a further dose increase maintained the current number of CSCs or decreased it. At the same time, the number of non CSCs significantly decreased with increasing doses of radiation exposure, as expected. Fractionated irradiation in vivo at a total dose of 10 Gy increased the relative amount of CSCs in most patients. The registered changes are an integral indicator of cell death, cell division delay immediately after irradiation, proliferation at a later time, possible dedifferentiation of non CSCs, etc. The exact contribution of each of them to the radiation-induced increase of the CSCs number is of considerable interest and requires further research.

[Influence of systemic photodynamic therapy on circulating tumor cells in peripheral blood of cancer patients].

The aim of this investigation is to study the number of circulating tumor cells (CTCs) in the blood of cancer patients after systemic photodynamic therapy (PDT) at different times and to assess apoptosis of these cells. The study group consisted of 19 patients with malignant tumors of epithelial origin at various stages (II-IV). CTC identification was performed with flow cytometry by immunophenotype Ep-CAM (CD326)+ CD45-. CTC apoptosis was identified by criteria of plasma membrane integrity and phosphatidylserine translocation on the outer surface of the membrane. Negative correlation between the CTC frequency and apoptotic death rate of these cells was found in patients before the treatment (R = -0.51, p = 0.03). CTC frequency gradually reduced during the first three days after PDT, and then it was maintained at the same level until the end of the follow-up (7 days). At the individual level, the effect of PDT depended on the frequency of CTCs before the treatment: the decrease in these cell frequency occurred significantly more often in the patients with an initially high frequency of CTCs than in other patients (p = 0.05). With the decrease in the CTC frequency, apoptotic death increased within 6 hours after the treatment and remained at the same level until the end of the follow-up period. The results demonstrate the efficacy of systemic PDT for elimination of tumor cells circulating in the peripheral blood of cancer patients with different localization of primary tumor and stage of disease.

Colonic microbiota alters host susceptibility to infectious colitis by modulating inflammation, redox status, and ion transporter gene expression.

Individuals vary in their resistance to enteric infections. The role of the intestinal microbiota in altering susceptibility to enteric infection is relatively unknown. Previous studies have identified that C3H/HeOuJ mice suffer 100% mortality during Citrobacter rodentium-induced colitis, whereas C57BL/6 mice recover from infection. The basis for their differences in susceptibility is unclear and has been mainly attributed to differences in host genetics. This study investigated the role of the intestinal microbiota in altering susceptibility to C. rodentium-induced colitis. When the feces of C57BL/6 mice were gavaged into antibiotic treated C3H/HeOuJ mice, the C57BL/6 microflora led to a complete reversal in mortality patterns where 100% of the C3H/HeOuJ mice survived infection. This protection corresponded with reduced colonic pathology and less systemic pathogen load and was associated with increased inflammatory and redox responses with reduced epithelial cell death. C3H/HeOuJ mice are normally susceptible to infection-induced dehydration due to defective expression of colonic ion transporters such as Dra, CA IV, and CA I; expression of these genes was normalized when C3H/HeOuJ mice were colonized with the C57BL/6 microflora. Together, these data reveal that the colonic microbiota play a critical role in protecting against intestinal infection by inducing proinflammatory and prooxidant responses that control pathogen load as well as ion transporter gene expression previously shown to prevent fatal dehydration. Protection of mice from lethal colitis was associated with higher levels of bacteria from Bacteroidetes. This study reveals that the microbiota is sufficient to overcome inherent genetic susceptibility patterns in C3H/HeOuJ mice that cause mortality during C. rodentium infection.

Characteristics of fatty acid composition of lipids in higher plant vacuolar membranes.

The fatty acid composition of vacuolar membrane lipids from plant storage tissues and their genesis have been studied. A high content of unsaturated fatty acids (up to 77%) was observed in lipids of these membranes. Linoleic acid prevailed in vacuolar lipids of carrot and red beet (54.2 and 44.2%, respectively). Linolenic acid prevailed in vacuolar lipids of garden radish and turnip (39.7 and 33.9%, respectively). Regarding saturated fatty acids, vacuolar lipids of garden radish, carrot, and red beet contained predominantly palmitic acid (up to 20-24%). Unsaturated fatty acids, petroselinic (C18: 1omega12), cis-vaccenic (C18: 1omega7), hexatrien-7,-10,-13-oic (C16:3omega3) and others, were observed in vacuolar lipids of roots. These acids are usually synthesized in chloroplasts, and their presence in vacuolar lipids can be associated either with the transport of metabolites to the vacuole, or with endocytosis during vacuolar formation in the plant cell. The specific features of fatty acid composition of tonoplast lipids apparently are closely related to the tonoplast unique fluidity and mobility required for running osmotic processes in the cell and for forming transport protein assemblies.

The structure of plant vacuolar membranes according to infrared spectroscopy.

The structure of the vacuolar membrane (tonoplast) was studied in red beet roots by IR spectroscopy. The vacuolar membrane was shown to be composed of highly ordered lipids which form regions of free liquid lipid bilayer loosely bound to integral proteins. The prevalence of polar lipids in the tonoplast is responsible for the high elasticity and fluidity of the membrane. The presence of alpha-tocopherol in the tonoplast membrane accounts for a high antioxidant activity of the membrane. Integral proteins are immersed into the liquid matrix of the lipid bilayer to a different extent. Examination of the temperature effect on the kinetics of the hydrogen-deuterium exchange in integral membrane proteins showed that the efficient energy of the hydrogen exchange activation was 24 +/- 4 kcal/mol at 19-40 degrees C and increased to 54 kcal/mol at 40-50 degrees C because of the thermal denaturation of proteins. The secondary structure of integral membrane proteins is characterized by a high content of alpha-helices (53%) which decreased to 8% after the extraction of lipids.

[The motivation for mental patients' attending industrial medical clinics]. / Motivatsiia poseshcheniia psikhicheski bol'nymi lechebno-proizvodstvennykh masterskikh.

In order to define the possibilities and approaches to the humanization and democratization of labour rehabilitation of mental patients, to reveal and consider individual requirements constituting each patient's personality, a study was made of the motives of attending the treatment and industrial shops (TIS) by them and preferable types of labour activities. Use was made of special formalized maps devised for the given study purposes. All the possible motives of attending TIS and preferable labour activities were registered in those maps as was an objective assessment of the output per man-hour. 221 patients were examined. The results of the examination were assessed in accordance with distribution of the patients by the clinical parameters. The hierarchy of the motives and preferences and output per man-hour depended on the patients' clinical characteristics. Still, it has been established on the whole that attending TIS, 91% were willing to work, 85% to change the environment, 70% to speed up the discharge, 65% to enhance the treatment efficacy, 45% to communicate with new people, 33% to earn money. 46% of the patients reported to work passively, under the influence of persuasions. 71% preferred performing collective work, 68% unsophisticated, 63% very simple work without any responsibility, 53% mobile, 51% sedentary, 49% physical, 32% complicated, 25% individual, 7% intellectual. Only 85% of the patients attending TIS worked. The highest productivity was recorded in patients with borderline mental disorders, the lowest in patients with actual psychotic disorders.