ANALYSIS OF THE BLOOD HYPERCOAGULATION RISK IN PATIENTS WITH ISCHEMIC ATHEROTHROMBOTIC STROKE DEPENDING OF THE VDR GENE POLYMORPHISMS.

OBJECTIVE: Aim: of our study was the analysis of the blood hypercoagulation risk in patients with ischemic atherotrombotic stroke depending of the VDR gene polymorphisms. PATIENTS AND METHODS: Materials and Methods: Blood of 170 patients with ischemic atherothrombotic stroke (IATS) and 124 healthy individuals (control group) was used for genotyping. Four polymorphisms (FokI, BsmI, ApaI, TaqI) of gene VDR were examined with PCR-RFLP methodology. Statistical analysis was performed by using SPSS-17.0 program. RESULTS: Results: Among patients with IATS who are carriers of the f/f genotype, FokI polymorphism of VDR gene by high thrombin time and a decrease in the rate of spontaneous fibrinolysis was registered. In individuals with the B/B genotype homozygous for the polymorphic variant, BsmI had significantly lower mean values of prothrombin and thrombin time and increased the rate of spontaneous fibrinolysis. The homozygotes for the A-allele ApaI polymorphism have 2.7 times higher risk of developing blood hypercoagulation than homozygotes for the a-allele was found. CONCLUSION: Conclusions: Biochemical signs of hypercoagulation syndrome among patients with IATS who are carriers of the f/f genotype of the FokI polymorphic variant and among B/B homozygotes of the BsmI polymorphic variant and homozygotes for the A-allele of the AрaI polymorphism of the VDR gene were registered.

The Link between ANRIL Gene RS4977574 Polymorphism and Common Atherosclerosis Cardiovascular Complications: A Hospital-Based Case-Control Study in Ukrainian Population.

Materials and Methods: 195 patients with ACS, 200 patients with LAS, and 234 control subjects were enrolled in this case-control study. Real-time PCR was used for ANRIL rs4977574 genotyping. SPSS software package (version 17.0, IBM, USA) was used for data analysis. Results: A significant association between rs4977574 polymorphism and the risk of atherosclerosis and cardiovascular complications was found under the recessive model regardless of adjustment for nongenetic risk factors (OR = 1.551; p = 0.025). Moreover, the link between rs4977574 locus and serum levels of total cholesterol (p = 0.021) and LDL (p = 0.022) was detected. A separate analysis in subgroups demonstrated the association of rs4977574 polymorphism with increased risk of ACS under the recessive model (OR = 1.501; p = 0.048). No relation between rs4977574 site and LAS development was revealed (p > 0.05). Conclusion: Obtained data suggested that ANRIL rs4977574-GG genotype can be a possible genetic marker for the development of atherosclerosis and cardiovascular complications in Ukrainian population.

Impact of MMP-9 Genetic Polymorphism and Concentration on the Development of Coronary Artery Disease in Ukrainian Population.

Coronary artery disease (CAD) is one of the leading causes of death in Europe. It is known that atherosclerosis is the primary risk factor of CAD development. MMP-9 is involved in all stages of atherosclerosis and thus may contribute to CAD emergence. To investigate the influence of MMP-9 on the (CAD) development 25 patients with intact coronary arteries (CA), 40 patients with acute coronary syndrome (ACS), and 63 patients with chronic coronary syndrome (CCS) were enrolled in the study. Real-time PCR was carried out for genotyping on the rs17567-polymorphic locus, and ELISA study was performed to measure the MMP-9 plasma concentration. It was found the lower risk of MI occurrence for AG-carriers (P a =0.023; ORa = 0.299, 95% CI = 0.106-0.848) in Ukrainian population.

Influence of LYS198ASN polymorphism of endothelin-1 gene on ischemic atherothrombotic stroke characteristics.

OBJECTIVE: The aim: Investigate the effect of Lys198Asn polymorphism of the EDN1 gene on ischemic atherothrombotic stroke characteristics. PATIENTS AND METHODS: Materials and methods: Venous blood of 170 patients with ischemic atherothrombotic stroke (IAS) and 124 patients without cerebrovascular pathology, who made up the control group, used for the study. Lys198Asn (rs5370) polymorphism of the EDN1 gene was determined by the polymerase chain reaction method followed by restriction fragment length analysis. Statistical analysis was performed using SPSS-17.0. The values of Р < 0.05 were considered reliable. RESULTS: Results: An association between the Lys198Asn polymorphism of the EDN1 gene and the IAS development was detected. For Asn/Asn genotype carriers, the risk of IAS developing is 4 times higher than that of homozygotes for the major allele. The association of this polymorphism with the arterial pool, whose atherothrombotic changes lead to the development of IAS, was found in individuals with BMI < 25 kg/m2. Lys198Asn polymorphism also affects the severity of IAS in persons with hypertension and non-smokers. CONCLUSION: Conclusion: The Lys198Asn polymorphism of the EDN1 gene influences some characteristics of ischemic stroke.

Association analysis between HOTAIR RS1899663 single nucleotide polymorphism and clear cell renal cell carcinoma development in Ukrainian population.

OBJECTIVE: The aim: to study the association between rs1899663-polymorphic variant of HOTAIR gene and clear cell renal cell carcinoma (CCRCC) development in Ukrainian population. PATIENTS AND METHODS: Materials and methods: whole venous blood from 101 Ukrainians with CCRCC (42 females and 59 males) and 100 control subjects (34 females and 66 males) were enrolled in the study. DNA extraction was performed using GeneJET Whole Blood Genomic DNA Purification Mini Kit (Thermo Fisher Scientific, USA). Polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) was used for HOTAIR rs1899663 genotyping. The Statistical Package for Social Science software (SPSS, version 17.0, Chicago, IL, USA) was used for all calculations. RESULTS: Results: It was found the lack of association between HOTAIR rs1899663 single nucleotide polymorphism and CCRCC emergence as well as tumor metastasis property in dominant, recessive, over-dominant and additive crude models of inheritance, as well after the adjustment for age, sex, smoking and excessive alcohol consumption (P > 0.05). CONCLUSION: Conclusions: No association was found between HOTAIR rs1899663-polymorphic variant and CCRCC development in Ukrainian population. Further studies with extended samples are required to validate these results.

Association Study between BGLAP Gene HindIII Polymorphism and Type 2 Diabetes Mellitus Development in Ukrainian Population.

Type 2 diabetes mellitus (T2DM) belongs to the diseases with hereditary predisposition, so both environmental and genetic factors contribute to its development. Recent studies have demonstrated that the skeleton realizes systemic regulation of energy metabolism through the secretion of osteocalcin (OCN). Thus, the association analysis between HindIII single nucleotide polymorphism of OCN gene (BGLAP) promoter region and T2DM development in Ukrainian population was carried out. 153 individuals diagnosed with T2DM and 311 control individuals were enrolled in the study. The genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The lack of association between BGLAP HindIII single nucleotide polymorphism (SNP) and T2DM development among Ukrainians was found. Further studies with extended groups of comparison are needed to confirm the obtained results.

The association of aрai-polymorphism of vitamin D receptor gene (VDR) with development of generalized parodontitis in Ukrainian population.

OBJECTIVE: Introduction: At present, it is believed that the genetic component is important in the pathogenesis of periodontitis. One of the candidate genes that are of major importance in the development of the disease is the vitamin D receptor gene (VDR). The association of its genetic polymorphisms, in particular Apal, with periodontitis in different populations of the world is proved. The aim: To study the association of the Apal-polymorphism VDR gene with the development of generalized periodontitis in the Ukrainian population. PATIENTS AND METHODS: Materials and methods: Patient genotypes were determined by polymerase chain reaction with subsequent analysis of restriction fragment length (PCR-RFLP) from buccal epithelium 116 patients with generalized periodontitis (GP) and 67 individuals of control group. Statistical analysis was performed by using SPSS-17,0 program. RESULTS: Results: As a result of the performed studies, it was shown that in the group of patients with GP, the ratio of homozygous for the main allele (a/a), heterozygote (a/A) and homozygote for the minor allele (A/A) was 26 (22,4%), 62 (53,4%), 28 (24,2%), and in control group - 25 (37,3%), 27 (40,3%), 15 (22,4%), respectively. The distribution of genotypes in the comparison groups was not statistically significant (P = 0,084). By the method of binary logistic regression in the framework of the additive inheritance model (a/A vs a/a), a reliable relationship of the genotype with the Apal-polymorphism of the VDR gene was established with the development of generalized periodontitis (Р=0,029). It was shown that in heterozygotes (a/A) the risk of GP in 2,208 (95% CI = 1,084-4,496) times is higher than in homozygotes of the main allele (a/a). After adjusting for age, sex, smoking habit, BMI, the reliability of these results was maintained (P = 0,030). CONCLUSION: Conclusions: The ApaI-polymorphism of the VDR gene is associated with the development of generalized periodontitis in the Ukrainian population.

[The association of LYS198ASN -polymorphism of endothelin-1 gene (EDN1) with development of arterial hypertension in ukrainian population].

OBJECTIVE: Introduction: Arterial hypertension is a multifactorial disease developing under the influence of environmental factors and is genetically determined. One of the genetic markers that is of primary importance in the disease development is endothelin-1 gene (EDN1). Today the association between the polymorphic variants of this gene, particularly Lys198Asn-polymorphism, and the development of arterial hypertension in different populations of the world has been proved. The aim: To study the association between the Lys198Asn-polymorphism of the endothelin-1 gene and the development of arterial hypertension in Ukrainian population. PATIENTS AND METHODS: Materials and methods: The genotypes were determined by the polymerase chain reaction method, followed by the analysis of the restriction fragment length (PCR-RFLP) in venousblood of 160 patients with arterial hypertension and 110 people in the control group. The statistical analysis was performed using SPSS-17.0. RESULTS: Results: As a result of genotyping, it was found that in the group of patients with arterial hypertension the ratio of homozygote of the major allele (Lys/Lys), heterozygote (Lys/Asn) and homozygote of the minor allele (Asn/Asn) was 74 (46.3%), 73 ( 45.6%), 13 (8.1%), while in control - 66 (60.0%), 41 (37.3%), 3 (2.7%) respectively. The distribution of genotypes in the experimental groups was statistically significant (χ2 = 6.66; P = 0.036). By the method of binary logistic regression within the dominant and additive model of inheritance, a reliable association between the genotype of the Lys198Asn-polymorphism of the ET-1 gene and the development of arterial hypertension was established. It was shown that carriers of minor allele (Lys/Asn+Asn/Asn) have a risk of arterial hypertension 1.7 (95 % CI = 1.066 - 2.851), and homozygotes Asn/Asn 3.9 (95 % CI = 1.016 - 9.566) times higher than people with Lys/Lys genotype. In addition, smoking patients with Lys/Asn and Asn/Asn- genotypes have a risk of arterial hypertension 2.6 (95% of SI = 1.224-5.488), and homozygotes of the minor allele (Asn/Asn) 7.3 (95% of SI = 1.295-41.639) times higher than the Lys/Lys homozygotes. CONCLUSION: Conclusions: Lys198Asn-polymorphism of the endothelin-1 gene is associated with the development of arterial hypertension in Ukrainian population. Carriers of minor allele (Lys/Asn+Asn/ Asn) have a risk of arterial hypertension 1.7, and homozygotes Asn/Asn 3.9 times higher than people with Lys/Lys genotype.

Positive Association between EDN1 rs5370 (Lys198Asn) Polymorphism and Large Artery Stroke in a Ukrainian Population.

There are a lot of convincing evidences about the involvement of endothelin pathway proteins in the pathogenesis of atherosclerosis and its fatal complications. In this study, the analysis of a possible association between EDN1 rs5370 and EDNRA rs5335 gene polymorphisms and the risk of large artery stroke (LAS) in a Ukrainian population was conducted. 200 LAS patients and 200 unrelated controls were enrolled in a case-control study. The polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP) was used for SNP genotyping. Our results revealed that EDN1 rs5370 polymorphism was associated with LAS development both before and after adjustment for atherosclerosis risk factors (sex, age, body mass index, arterial hypertension, type 2 diabetes mellitus, and smoking). The risk for a LAS incident in rs5370-T allele carriers was 1.6 times higher (CI = 1.066-2.403; P = 0.020) than in subjects with the GG genotype. No link between EDNRA rs5335 and LAS risk in a Ukrainian population was found. The present study indicated that EDN1 rs5370, but not EDNRA rs5335, can be the strong genetic predictor for LAS development in a Ukrainian population.

[The association between enpp1 rs997509 polymorphism and type 2 diabetes mellitus development in ukrainian population].

OBJECTIVE: Introduction: Genome-Wide Association Studies have identified a large number of polymorphic loci associated with type 2 diabetes mellitus (T2DM). Ectonucleotide Pyrophosphatase/Phosphodiesterase 1 (ENPP1) gene is one of the candidate genes which have primary importance in T2DM development. Several studies revealed the association between ENPP1 polymorphisms, including rs997509, and T2DM, obesity, insulin resistance and metabolic syndrome in different populations. The aim: To test the association between ENPP1 rs997509 polymorphism and T2DM development in patients with different risk factors in the Ukrainian population. PATIENTS AND METHODS: Materials and methods: Venous blood of 317 unrelated T2DM patients and 302 healthy volunteers was used for analysis. ENPP1 rs997509 genotyping was performed using PCR-RFLP (polymerase chain reaction with following restriction fragment length polymorphism analysis) method. RESULTS: Results: Our results revealed that ratio of C/C homozygotes, C/T heterozygotes and T/T homozygotes between case and control groups was significantly different (89.0 % , 11.0%, 0 % vs 94.4 %, 5.6 %, 0 %, P = 0.015). It was shown that risk of T2DM development in T allele carriers is significantly higher compared to C/C homozygotes (OR = 2.086; P= 0.027). Herewith the risk increased in heterozygotes with BMI ≥ 25 kg/m2 (OR = 2.223, P=0.031) and obesity (OR = 3.230; P = 0.023). CONCLUSION: Conclusions: ENPP1 rs997509 polymorphism is associated with T2DM development in Ukrainian population.

The histological and electron microscopic study of the parotid salivary gland in dehydrated rats of different ages.

OBJECTIVE: Introduction: Water-salt metabolism disorders is one of the main factor of salivary gland pathology development. The aim: To study the morphological structure of the parotid salivary gland of young, mature and old rats at micro- and ultrastructural levels under water deprivation. PATIENTS AND METHODS: Materials and methods: The experiment was carried out on thirty six laboratory male rats of different ages (young, mature and old). The rats of the control group received normal volume of drinking water. The rats of the experimental group were deprived of water for 6 days. Light microscope "OLYMPUS" and transmission electron microscope JEM-1230, (JEOL, Japan) were used for structural analysis. RESULTS: Results: Obtained results revealed increasing numbers of vacuoles in the serous cells, the enlarged cisterns of endoplasmic reticulum and Golgi apparatus tubules, the condensed chromatin and the nuclei with significant invaginations in parotid gland of the rats of all age groups. The area of the acinuses more changed in young rats, the decrease was 34.61 % (P = 0.007). The internal diameter of capillaries most decreased in the dehydrated old rats by 23.76 % (P = 0.009) in comparison with all study groups. CONCLUSION: Conclusions: Water deprivation brings about the structure changes of the parotid gland at micro- and ultrastructural levels the intensity of which depends on the age of animals. The most dramatic changes have occurred in young and old rats.

[Role of c+70g single nucleotide polymorphism of endothelin receptor a gene in the development of ischemic atherothrombotic stroke].

INTRODUCTION: Endothelial dysfunction is the basic pathogenic development mechanism of ischemic atherothrombotic stroke (IAS). One of the reasons for changes of endothelin structure and properties and its receptor can be genetic polymorphism of their genes. AIM: Study of C+70G polymorphic variant associations of endothelin receptor A gene (EDNRA) with IAS development became the goal of research. MATERIAL AND METHODS: Venous blood of 170 patients with IAS and 124 persons without cerebrovascular pathology was used for the research. The groups did not differ in the ratio of two sexes (P = 0,294 for the ?2-test), but the average age of the first group (76,7 ± 0,93 years) was significantly higher than that of the second one (P < 0,001). Determination of allelic variant of EDNRA gene by C+70G polymorphism was carried out using method of polymerase chain reaction with further analysis of restriction fragment length. RESULTS: As a result of the conducted genotyping, it has been discovered that homozygote correlation by the main allele (CC), heterozygotes (CG) and homozygotes by minor allele (GG) does not differ authentically in patients with IAS and control group (24.1 %, 57.6 %, 8.2 % against 29.0 %, 50.0 %, 21.0 %; ? = 0.426). It is shown that association of the studied genetic marker with the scope of brain damage, localization of atherothrombotic process, course severity, recurrence and neurological manifestations of IAS. CONCLUSIONS: There is no association between the C+70G polymorphism of the EDNRA gene and the development of IAS and its clinical characteristics.

G-1639A but Not C1173T VKORC1 Gene Polymorphism Is Related to Ischemic Stroke and Its Various Risk Factors in Ukrainian Population.

Vitamin K epoxide reductase complex subunit 1 (VKORC1) is integral 163-amino acid long transmembrane protein which mediates recycling of vitamin K 2,3-epoxide to vitamin K hydroquinone and it is necessary for activation of vitamin K-dependent proteins (VKDPs). Herein, the association between G-1639A (rs9923231) and C1173T (rs9934438) single-nucleotide polymorphisms (SNPs) of the VKORC1 gene and ischemic stroke (IS) was tested in Ukrainian population. Genotyping was performed in 170 IS patients and 124 control subjects (total 294 DNA samples) using PCR-RFLP (polymerase chain reaction with following restriction fragment length polymorphism analysis) method. Our data showed that G-1639A but not C1173T polymorphism was related to IS, regardless of adjustment for age, sex, body mass index, smoking status, and arterial hypertension. The risk for IS in -1639A allele carriers (OR = 2.138, P = 0.015) was higher than in individuals with G/G genotype. Haplotype analysis demonstrated that -1639G/1173T and -1639A/1173C were related to increased risk for IS (OR = 3.813, P = 0.010, and OR = 2.189, P = 0.011, resp.), while -1639G/1173C was a protective factor for IS (OR = 0.548, P < 0.001). Obtained results suggested that -1639A allele can be a possible genetic risk factor for IS in Ukrainian population.