We used quantitative real-time PCR method to analyse mtDNA copy number in a random subsample (n=996; 358 men aged 66,31±7,24 years; 468 women aged 67,62±7,1 years) selected from a population cohort (n=9 630) examined at baseline in international project HAPIEE in Novosibirsk, Russia, in 2003-2005. The participants were re-examined after 12 years in 2015-2017. The average relative number of mtDNA copies in peripheral blood leukocytes was greater in women than in men, independently of age and smoking (p=0,001). mtDNA copy number was inversely correlated with age both in men (p=0,005) and women (p<0,001). In age adjusted analysis, mtDNA copy number was inversely associated with waist, hip and heart rate in both sexes. In addition, mtDNA copy number in women was inversely associated with triglycerides and glucose, aterogenity index and positively with HDL cholesterol. In men, mtDNA copy number was positively associated with physical activity. The age-adjusted mean of mtDNA copy number among male never-smokers was greater than in smokers (p=0,003), and the mean mtDNA copy number was lower in women with diabetes than in women without diabetes (p=0,005). In both sexes, subjects with baseline history of hypertension had lower mtDNA copy number after 12-year follow-up than those without hypertension (p=0,05). This broadly supports the hypothesis that mtDNA copy number may act as biomarker of ageing.

Aim. To analyze differential expression of metalloproteases genes, involved into the processes of stabilization/destabilization of atherosclerotic plaque, with the method of full genome sequencing of RNA, and to evaluate the level of metalloproteases in homogenates of plaques of various types by immune enzyme assay method (IEA).Material and methods. The study has been conducted on the specimens of atherosclerotic plaques of patients aged 45-65 y.o., inhabitants of Western Siberia with angiographically proven coronary atherosclerosis and no acute coronary syndrome, with stable angina II-IV functional class. Specimens collection from the plaques was done during an operation if there were intraoperational indications. Histology performed. In intima/media homogenates by IEA method, with BCM Diagnostics assays the levels of destructive markers were measured: MMP-1, MMP3, MMP-7, MMP-9, TIMP-1 on the Multiscan EX (Thermo Fisher Scientific, USA). Libraries preparation for full genomic sequencing of RNA was done with Illumina’s TruSeq RNA Sample Preparation Kit (Illumina, USA). Expression profile in tissues was done on HiSeq 1500 (Illumina, USA).Results. There are differences in expression of the genes MMP2, MMP7, MMP8, MMP9, MMP12, и MMP14 in different types of plaques. There was 8 times higher significant raise in increase of the expression level of ММР9 (p<0,001) in unstable plaque of dystrophic-necrotic type. Study by IEA of MMP-7 content, which is an activator of pro-MMP-9, as well as the content of MMP-9 itself, showed their increased levels in unstable plaques comparing to fatty streaks (1,5 and 2,4 times) and young stable plaques (1,4 and 2,1 times).Conclusion. For the gene ММР9 there were significant differences obtained, of expression levels in stable atherosclerotic fibrous plaque and unstable plaque of dystrophic-necrotic type. With the IEA it was found that fatty streaks and young stable atheromas of coronary arteries have an increased concentration of MMP-3 and decreased activity of tissue inhibitor of metalloproteases. In unstable plaques with the tendency to rupture/ulceration there are increased levels of MMP-1, MMP-7, MMP-9.

Genotyping of TRPV1 and TRPA1 genes encoding thermoreceptors in the populations of the Altai-Sayan region and the Far East was conducted. The sample consisted of 15 populations comprising 1482 individuals. The analysis of TRPV1 rs222747 demonstrated that the frequency of M315I was closest to East Asian populations only in Nanais and Koryaks (56 and 64%, respectively). Siberian Tatars, Yakuts, and Evenks were closest to European populations. All populations of the Altai-Sayan region reported an intermediate position between the Caucasoids and the Eastern Mongoloids on the basis of the frequency of M315I. No deviations from the Hardy–Weinberg distribution were observed. The observed heterozygosity exceeded the expected one in eight populations. The analysis of TRPA1 rs13268757 revealed that Chukchi, Yukaghir, Koryak, Tuvinian, Southern Altaian, and Telengit populations were closest to the East Asian populations on the basis of the frequency of R3C substitution (3–7%). At the same time, populations of Siberian Tatars, Nanais, Evenks, Yakuts, Shorians, Khakases, and Kazakhs were intermediate between the Caucasoids (18–23%) and the Mongoloids from East Asia (3–7%) on the basis of the frequency of this polymorphism. A deviation from the Hardy–Weinberg distribution was detected only in the Yukaghir population. The observed heterozygosity was higher than the expected one in nine populations. A trans-association of TRPA1 and TRPV1 gene polymorphisms was carried out in 14 populations via regression analysis. A negative correlation of–0.545 was determined, the number of degrees of freedom ( df ) was 13, and the P -value was 0.048. The data obtained indicate that the analyzed polymorphisms are correlated, which confirms an earlier conclusion of the TRPA1 -dependent inhibition of TRPV1 function. The results may evidence in the co-evolution of analyzed genes.

Leukocyte telomere length (LTL) associations with age, sex, and risk factors for age-related diseases have been studied in Russian people of preretirement and retirement age. This parameter has been determined by quantitative real-time PCR in 398 men (56.3 ± 7.2 years) and 365 women (56.6 ± 7.1 years) selected from a population sample of 45 to 69-year-old residents of the Oktyabrskii and Kirovskii raions of Novosibirsk (9400 people). The sample was formed in the course of the international project HAPIEE. Telomere length has been found to correlate with age (r =–0.159, p < 0.001) and the waist: hip ratio (WHR) (r =–0.107, p = 0.003). The average LTL in women is significantly greater than in men, p = 0.031. In men, LTL correlates with body mass (r = 0.140, p = 0.005) and waist size (r = 0.111, p = 0.027). In women, there are inverse correlations of LTL with waist size (r =–0.127, p = 0.015) and WHR (r =–0.141, p = 0.007). Leukocyte telomere length inversely correlates with the amount smoked (r =–0.121, р = 0.024). It directly correlates with age, smoking, and a variety of phenotypical traits. In men with family histories of malignancies, LTL is greater than in men without such histories.

Aim. To search and study the association of some candidate genes polymorphysms of various cardiovascular diseases and sudden cardiac death in men.Material and methods. The sudden cardiac death group (SCD) is collected by the WHO criteria with suddenly died men underwent court-medicine expertise (n=274). Control group was matched by the age and gender from the DNA bank of HAPIEE and MONICA trials. Genotyping of groups was done according to polymorhysms SCN5A, rs187238 gene IL-18, rs1799864 gene CCR2, rs3864180 gene GPC5, rs1799983 gene eNOS, rs2228314 gene SREBF-2, rs1800588 gene HL, rs10757278, rs1333049 with methods of PDRF and realtime PCR.Results. No significant differences found between the SCD group and control group by the prevalence of genotypes SCN5A, rs187238 gene IL-18, rs3864180 gene GPC5, rs1799983 gene eNOS, rs1800588 gene HL.In SCD group the decrease of homozygotes by CC polymorphism rs2228314 found in gene SREBF-2 and the decrease of heterozygotes GC comparing to control group (HR=4,074, 95% CI 1,843-9,002, р=0,0002; HR=0,442, 95% CI 0,302-0,647, р=0,0001, reap.). In SCD group carriers of GG genotype polymorphism rs10757278 and CC poly morphism rs1333049 are significantly more prevalent than in control group (HR=1,814, 95% CI 1,159-2,839, р=0,011; HR=1,744, 95% CI 1,104-2,754, р=0,019, resp.). The part of GA carriers of polymorphism rs1799864 gene CCR2 is more prevalent in SCD group comparing to control (HR=1,558, 95% CI 1,051-2,308, р=0,029).Conclusion. Polymorphisms rs10757278, rs1333049, rs2228314 gene SREBF-2, rs1799864 gene CCR2 are associated with sudden cardiac death in men.