FREQUENCY OF VKORC1 AND CYP2C9 GENES POLYMORPHISM IN ABKHAZIAN POPULATION.

The aim of the research was to study the frequency of VKROC1 and CYP2C9 genes different alleles for healthy donors and for patients with thrombosis, in Abkhazian population and to reveal the interdependence of the studied genes products in the treatment of thrombosis with warfarin. Warfarin is an anticoagulant, causing the inactivation of the VKORC1 gene product, which is one of the clotting factors. The protein product of CYP2C9 gene is involved in the metabolism of warfarin. Genotyping of blood samples for studied genes alleles was carried out using a tube scanner (ESE Quant Tube Scaner), allowing to identify SNPs. With the highest frequency in the studied group of healthy donors of Abkhazian population, by VKROC1 gene found Heterozygous (AG genotype) (74,5 %). The distribution of homozygous of "wild" (GG) and mutant genotype (AA) accounted for 13,5% and 11,8%, respectively. In the group of patients with Thrombosis, wild-type homozygotes accounted for 32.5%, which is significantly high compared to the control group. The percentage of heterozygotes was significantly lower than in the control group and accounted 56,25%. as for the homozygous mutant genotype, it was practically the same as in control group (11,2%). Regarding the rate of polymorphic variants of the CYP2C9 gene, quite large differences between diseased and healthy individuals were detected according to some of them. CYP2C9 *1/*1 genotype (wild- type homozygote) was observed in 32.9% of healthy individuals, while the same genotype was detected in only 14.5% of patients with thrombosis. The percentage of CYP2C9 *1/*2 genotype was slightly different between healthy and thrombotic subjects and corresponded to 27.5% in healthy individuals and 30.4% in thrombotic patients. CYP2C9 *1/*3 genotype accounted for 16.1% in healthy individuals. The mentioned indicator was significantly different from the similar indicator of patients with thrombosis, which corresponded to 24.1%. The largest difference between the percentages was observed according to the CYP2C9 *2/*3 (mutant heterozygote) genotype. In healthy individuals, this rate corresponded to 40.3%, and in thrombotic individuals - 11.4%. The CYP2C9 *2/*2 genotype was not observed in any of the study groups, while the percentage of CYP2C9 *3/*3 (mutant homozygous) individuals did not differ and amounted to 1.6% (in healthy individuals) and 1.2% (in thrombotic patients). VKORC1 and / or CYP2C9 genes polymorphisms are presented in a number of clinical dosing algorithms and in prospective clinical trials. In conclusion, it should be noted that the present work revealed a significant variability of genotypes between the groups of patients with thrombosis and healthy individuals, in Abkhazian population. The results obtained in determining the polymorphic variants of the VKORC1 and CYP2C9 genes, studied by us, should be taken into account when using algorithms to determine the optimal dosage for warfarin treatment in thrombotic individuals of the Abkhazian population, both during treatment and for the prevention of thrombosis.

THE SIGNIFICANCE OF CIRCULATING SURFACTANT PROTEIN D AND DYSLIPIDEMIA IN CHRONIC OBSTRUCTIVE PULMONARY DISEASE, CORONARY HEART DISEASE AND THEIR COMBINATION.

As it is widely known, cardiovascular diseases represent one of the leading causes of mortality. In the coexistence of chronic obstructive pulmonary disease (COPD), the mortality rate is increasing as well. The research conducted reviews the effect of surfactant protein D on the coronary heart disease and chronic obstructive pulmonary disease.The cohort of 90 patients was included in the study. The patients were divided into five groups: group I (the patients with coronary heart disease - CHD); group II (the patients with dyslipidemia); group III (the patients with COPD); group IV (the patients suffering from the combination of CHD and COPD); group V (control group). The laboratory (dyslipidemia - through enzymatic method) as well as the instrumental methods (echocardiography, spirometric examination) were applied in the study.In consequence of the statistical processing of materials within the study, the relatively high level of surfactant protein D (SP-D) values determined in groups was revealed in group II (the patients with dyslipidemia), SP-D-25.9±19.6 ng/ml. Similar data were identified in practically healthy individuals - control group (group V). Performing the comparative analysis of data, the statistically high value of anti-atherogenic HDLC (52.5±10.5 mg/dl) was observed in group II (the patients with dyslipidemia) compared to group IV (p=0.002). It is worth noting that in the patients of group IV (COPD+CHD) the quantitative HDLC was statistically lower (43.6±6.5mg/dL) in comparison with the data of all other groups: (group I - HDLC-51,6±6,8; group II -52.5±10.5; group III-50.7±9.5; group V - 50.3±8.4) p1-4<0.001; p2-4=0.002; p3-4=0.005; p4-5=0.02. Considering the above-mentioned data, we can draw the conclusion that along with the elevated level of circulated SP-D, the low HDLC value can be utilized as a marker of the severe course of disease in relation to COPD as well as CHD.The high level of SP-D, which does not have the significant atherogenic effect (due to the presence of high HDLC), was detected in individuals with isolated dyslipidemia. The study has established that in the presence of CHD, in line with the increase in atherogenic lipoproteins and relatively low HDLC value, there is detected the high level of SP-D, which is correlated with lung function indices (FEV 1 and FVC). And in case of the coexistence of COPD and CHD where dyslipidemia, the elevated level of SP-D and corresponding changes in pulmonary function tests are manifested, there were identified the high risks of severe course of disease, the development of heart failure and mortality.

CORRELATION OF THYROID AUTOIMMUNITY WITH ATHEROSCLEROSIS EVALUATION IN HASHIMOTO'S THYROIDITIS.

The relationship between subclinical hypothyroidism (SH) and Atherosclerotic (At) cardiovascular diseases (CVD) has been one of the most popular topics but causal connection between Hashimoto thyroiditis (HT), lipid profile and follicular epithelial molecular biology is controversial. We investigated 3 groups of patients (group I - HT, group II - HT+At, group III - At). All laboratory tests for thyroid function and lipid profile detection were used according to international guideline recommendations, coronary and femoral arteries intima-media thickness (IMT) were tested by high-resolution ultrasonography, thyroid gland histology and immunohistochemistry carried out by p63 and S100 protein expression control. The statistical analysis was performed using Microsoft Excel 7.0, SPSS-20 version, Mann-Whitney U-test and Pearson's correlation. Comparisons between groups and factors were made using Multiple Linear Regression model. With the results obtained, dyslipidemia and the diastolic hypertension accelerate the hypothyroidism in HT+At group to predispose carotid and femoral arteries IMT. TSH and anti-TPO antibody levels are directly linked to the cardiovascular complications. Biomarkers S100 and p63 data show negative feedback effects of hypercholesterolemia on the high morphological risk features in Hashimoto parenchyma, which may partially explain the significant trend and pathobiological link of HT with Papillary thyroid carcinoma.

[FREQUENCY OF POLYMORPHISM OF VKORC1 AND CYP2C9 GENES IN TWO REGIONS OF GEORGIA].

The aim of the research was to study the frequency of VKROC1 and CYP2C9 genes different alleles for healthy donors and for patients with thrombosis, in two regions of Georgia - in Samegrelo and in Tbilisi and to reveal the interdependence of the studied genes products in the treatment of thrombosis with warfarin. Warfarin is an anticoagulant, causing the inactivation of the VKORC1 gene product, which is one of the clotting factors. The protein product of CYP2C9 gene is involved in the metabolism of warfarin. Genotyping of blood samples for studied genes alleles was carried out using a tube scanner (ESE Quant Tube Scaner), allowing to identify SNPs. In the studied group of patients with thrombosis from Samegrelo region the wild-type homozygotes by the gene VKORC1 were - 90%; heterozygotes - 10%; mutant homozygotes have not met at all. In the studied group of patients with thrombosis from Tbilisi, also predominated homozygous wild type (60%); heterozygotes were - 40%; mutant homozygotes were not met. The genotypes of healthy donors fromTbilisi does not differed from the same indicator of of Samegrelo (homozygous "wild" AA - 37%; genotype AB - 47%; and mutant genotype - BB - 16%). In patients with thrombosis, from Samegrelo, wild-tipe homozygotes and heterozygotes by CYP2C9 gene were almost the same rate (51% and 49% -, respectively); mutant homozygotes were not revealed. In patients from Tbilisi, the frequency of wild-type homozygotes was 70%, heterozygotes and mutant homozygotes was 20% and 10% - respectively. The ratio of the frequencies of CYP2C9 gene alleles in healthy donors from Tbilisi and Samegrelo is not different - wild-type homozygotes - 77%; heterozygotes - 23%; mutant homozygotes in both regions were not met. VKORC1 and / or CYP2C9 genes polymorphisms are presented in a number of clinical dosing algorithms and in prospective clinical trials. It is revealed the significant variation of genotypes in patients with thrombosis (in both studied regions), which indicates the importance of as in treatment process, as well as for the prevention of thrombosis.

[Genomic instability in atherosclerosis].

A comparative study of the level of genomic instability, parameters of quantitative and structural mutations of chromosomes (aberration, aneuploidy, polyploidy) in lymphocyte cultures from patients with atherosclerosis of age 80 years and older (control group - 30-35 years old) was conducted. The possibility of correction of disturbed genomic indicators by peptide bioregulators - Livagen (Lys-Glu-Asp-Ala) and cobalt ions with separate application or in combination was also studied. Control was lymphocyte culture of two healthy respective age groups. It was also shown that patients with atherosclerosis exhibit high level of genomic instability in all studied parameters, regardless of age, which may suggest that there is marked increase in chromatin condensation in atherosclerosis. It was also shown that Livagen (characterized by modifying influence on chromatin) separately and in combination with cobalt ions, promotes normalization of altered genomic indicators of atherosclerosis in both age groups. The results show that Livagen separately and in combination with cobalt ions has impact on chromatin of patients with atherosclerosis. The identified protective action of Livagen proves its efficacy in prevention of atherosclerosis.

[Interrelation of lipid status with staging of breast cancer].

The lipid status of 30 women (38-64 years) with breast cancer (stages I-II A, B) was investigated. Selection and distribution of patients in groups occurred on the basis of the accepted classification by the size, invasivity and presence of metastases. The main changes in the lipid formula of blood included increased level of total cholesterol and low density lipoproteins cholesterol. The relationship between dislipoproteinemia and invasivity were reveald. Thus, these changes can be considered as predictor of growth and distribution, and correction of lipid homeostasis (dietary and medicamentous) as a primary factor of secondary preventive maintenance of breast cancer.

[Genetic aspects of longevity and aging].

To clarify the role of genetic factors in aging and longevity some clinical and genealogical characteristics of different age groups (54 patients of both sexes) were studied; 34 of them were long livers (>90y) and 20 - the elderly (70-90y). Clinical and laboratory investigations were provided according to WHO experts recommendations. Total content of Cholesterol and Triglycerides was assessed by the enzyme-measuring method. Based on the genealogical questionnaire methods, the data concerning morbidity, mortality, health status and CVD were collected about 206 people (in total) including the parents and siblings of the patients. The long-livers' fathers were found to live longer than the fathers of the elderly that confirms the idea of paternal-line inheritance of life longevity. The male longlivers' fathers live 10 years longer, than female longlivers' fathers (P<0,05). Maternal line doesn't influence on the longevity. We also noticed genealogical relation between health status of longlivers (absence of cardiovascular diseases) and genealogical anamnesis of their parents. The stability of lipid metabolism was observed in longlivers and the elderly that was confirmed by the low amount of total Cholesterol being lower in the longlivers. In both groups the correlation between the total Cholesterol content and the percentage of CVD cases was observed. Proceeding from the above, we can confirm the important role of genetic factors to reach the long age.

[The role of oxidative metabolism disturbance in the development of NO-related endothelial dysfunction during chronic hearth failure].

The aim of the work was to establish the oxidative metabolism changes and NO data in Chronic Hearth Failure (HF). 52 patients were included in the investigation, among them 37 patients with CHD and chronic HF (II-IV functional class by NIHA) and 17 without it (control group). For revealing of organism redox-status (ceruloplasmine, Fe3+-transfferine, Mn2+, methemoglobine) the blood paramagnetic centers was studied by electron paramagnetic resonance method. For revealing of blood free NO, the diethyldithiocarbamat (SIGMA) was used. In chronic HF the oxidative process intensification and organism compensate reaction reduction with low Fe3+-transferine levels, increased Mn2++, methaemoglobin and inactivation of erythrocytes membranes adrenergic receptors were revealed. In chronic HF the accumulation of reactive oxygen levels provoke NO transformation in peroxynitrote with following decreases of blood free NO and develop the endothelial dysfunction.

[Morphological and functional markers of the histo-hematological barrier in experimental hypercholesterolemia]. / Morfofunktsional'nye pokazateli gistogematicheskogo bar'era pri eksperimental'noi giperkholesterinemii.

Results of the experiments carried out have shown that in experimental hypercholesterinemia it is disordered microcirculation together with mediated changes in permeability of the histohematic barrier, lipoprotein-rich plasma in the pericapillar space, with the organ parenchyma trophicity getting compromised that are primarily responsible for the presence of dystrophy and fibrosis of organs. Unidirectionality and systemic pattern of the reaction is evidence of a generalized character of affections of the terminal vascular bed in response to exposure to one of the risk factors for atherogenesis, viz, exogenous cholesterin and dyslipidemia.

[Effects of exogenous cholesterol on the functional morphology of blood formed elements in atherogenesis dynamics]. / Vliianie ékzogennogo kholesterina na funktsional'nuiu morfologiiu formennykh élementov krovi v dinamike aterogeneza.

Results of clinical and experimental studies showed that exogenous cholesterol is a factor of initiation of athero- and thrombogenesis that under conditions of failure of compensatory reactions are manifested by generation and progression of illness. In the control group of observation, activation of leukocytes in indicative of reciprocal reactions of cell-bound immunity directed to elimination of atherogenic lipoproteids whereas in patients with ischemic heart disease, the cholesterol load apparently provokes cell type effector reactions leading to damage to and necrosis of cells. In healthy subjects, chemotaxis and phagocytosis become activated while in sick persons they get supressed. We can well believe that there takes place fixation of an antigen (in the given case--apo-beta lipoproteins) on macrophages and polymorphonuclear leukocytes with subsequent cascade to follow such as activation of lipooxygenase, active forms of oxygen, superoxydes, cytokins, as well as production of elastase and collagenase, which fact in the presence of a high degree inter-cellular cooperation results in formation of a vicious circle. The findings secured corraborate etiological significance of exogenous cholesterol and saturated fatty acids as a starting-point of atherogenesis.