In recent years levels of a number of inflammatory markers namely C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor (TNF) etc. are measured for the purpose of postinfarction risk evaluation. Dynamics of inflammatory markers concentrations can reflect processes occurring in atherosclerotic plaque and coronary arteries. Concentrations of inflammatory markers depend particularly on genetic factors affecting transcription levels of individual genes. This data suggest that genotypes which determine increased inflammatory markers levels in blood can increase risk of unfavorable events after myocardial infarction. STUDY PURPOSES: Analysis of influence of allelic polymorphisms C1444T of CRP gene (rs1130864), G(-174)A of IL6 gene (rs1800795), A(-308)G of TNF gene (rs1800629), G252A of LTA gene (rs909253), (-509) of TGFB1 gene (rsl800469) and delta32 (w/d) of CCR5 gene (rs333) on development of cardiac unfavorable events in Russian patients with MI during two years follow-up. 211 Russian patients were included (52.3+/-10.3 years), 160 men (50.1+/-10.6 years) and 51 women (55.2+/-10.1 years). After two years of follow-up patients were examined in hospital, or telephon call occurred for determination of patient's condition or end point assessment. The end points were cardiac death, recurrent MI, recurrent hospitalization with unstable angina or stroke, CABG or PTCA performing. The genotyping was performed by methods based on polymerase chain reaction (PCR): PCR-SSP and PCR-RFLP. Analysis revealed association of allele T (p=0.036, OR=1.6, 95%CI: 1.052.6) and of allele T carriage (genotypes CT+TT) (p=0.046, OR=1.9, 95%CI: 1.053.6) of polymorphism C1444T of CRP gene with unfavorable events development. Analysis of survival rate by Kaplan-Meier estimation showed that cumulative part of patients without unfavorable events was significantly lower among allele T carriers than among carriers of genotype C/C of polymorphism C1444T CRP. Allele A of polymorphism A252G of LTA gene was also associated with unfavorable events risk (p=0.034, OR=1.96, 95%CI: 1.073.06). There was no association of polymorphisms delta 32 (w/d) of CCR5 gene, A(-308)G of TNF gene, G(-174)C of IL-6 gene, C(-509)T of TGFB1 gene with unfavorable events development.

Multiple sclerosis (MS) is a chronic inflammatory autoimmune disease of the central nervous system (CNS). Proteins of the immune system, as well as proteins that are involved in the infiltration of activated immune cells in the CNS, play an important role in the pathogenesis of MS. We investigated the association and linkage with MS of the following immune-system genes polymorphisms: HLA-DRB1,CTLA4,TGFB1,IL4,CCR5 andRANTES, as well as of the matrix metalloproteinase 9 (MMP9) and tissue inhibitor of metalloproteinase 1 (TIMP1) genes polymorphisms. For this purpose we used the transmission disequilibrium test (TDT). The group investigated was comprised of 100 nuclear families of Russian ethnicity, each consisting of an affected offspring and his nonaffected parents. It was found that HLA-DRB1*15alleleandMMP9*-1562C allele were transmitted from healthy heterozygous parents to affected children more frequently than alternative alleles (p = 0.02 andp = 0.04, respectively). Another family-based method, AFBAC (affected family-based control), showed MS association with HLA-DRB1*15, but not with theMMP9*-1562C allele.

Objective: Comparison of distribution of alleles and genotypes of polymorphic loci of RAAS genes: CYP11B2 (C-344T), REN (C-5434T, C-5312T, and A BglI G), CMA (G–1903A), ACE (I/D) and AT2R1 (A1166C) and their combinations in patients with low-renin forms of arterial hypertension (AH). Design and Method: 87 hypertensive pts aged 50 ± 13 years with low-renin forms of AH were studied. PAC and PRA were determined in the supine position and following 4-hour upright posture. Complex statistic analysis of differences in allele and genotype patterns distribution between different forms of low-renin AH was carried out by means of recently developed by our group software APSampler, which used a Bayesian Markov chain Monte Carlo-based method and provided an independent validation by means of Fisher's exact test. The differences were considered significant at p values < 0.05. RESULTS: According to clinical data and comparison with PAC, PRA and instrumental analysis pts were divided into 3 groups: group Ia (n = 41) with primary hyperaldosteronism (HA) associated with adenoma; group Ib (n = 18) with idiopathic HA -adrenocortical hyperplasia; combined group I included all 59 HA cases; and group II (n = 28) cases with low-renin essential hypertension with a normal PAC and the absence of structural adrenal changes. The analysis of carriership of allele and genotypes combinations showed significant differences between groups Ia, Ib and II. A comparison of groups II and Ia identified assemblage associated with predisposition only to primary HA: (–344T) CYP11B2 + BglI G REN + I ACE (p = 0.009, OR = 5.4 (95% CI 1.4 to 21.2)) and a comparison of groups II with group Ib identified assemblage as predisposing only to IHA: (–5312 T) REN + D ACE (p = 0.004, OR = 10.7 (95% CI 1.9 to 60.1)). Conclusions: Assemblage (–344T) CYP11B2 + BglI G REN + I ACE is factor of genetic predisposition only to PHA and assemblage (–5312 T) REN + D ACE - only to IHA, thereby indicating that these assemblages are risk factors for different forms of low-renin HA.

Провоспалительные цитокины интерлейкин-6 (IL-6), интерферон- (IFNg) и фактор некроза опухолей (TNF), известные как участники воспаления, играют важную роль в патогенезе рассеянного склероза. Исходя из опубликованных данных о влиянии полиморфизмов G(–308)A гена TNF, A(+874)T гена IFNG и G(–174)C гена IL-6 на продукцию этих цитокинов, мы изучали связь между полиморфизмом этих участков и развитием рассеянного склероза. Сцепление и ассоциацию аллелей указанных генов с рассеянным склерозом анализировали, используя тест неравновесной передачи аллелей (transmission disequilibrium test, TDT). В группе из 104 ядерных семей русской этнической принадлежности (больной и его здоровые родители) обнаружено, что аллель TNF*(–308)A чаще передается больным детям от здоровых гетерозиготных родителей (p = 0.01). Сцепления/ассоциации с рассеянным склерозом аллелей генов IFNG и IL-6 не выявлено. Таким образом, полученные данные свидетельствуют об участии гена TNF в развитии предрасположенности к рассеянному склерозу у русских.

Frequencies of the carriership of alleles and genotypes of functionally important polymorphous loci of some inflammation genes (proinflammatory cytokine genes IL-6, LTA, and TNF; anti-inflammatory cytokine gene TGFB1; and CC chemokine receptor 5 gene CCR5) were analyzed in 199 ethnic Russian patients with myocardial infarction (MI) and in a control group of 142 persons of the same ethnic descent. Complex analysis by the APSampler algorithm revealed associations of MI with the carriership of all polymorphic variants either regarded as individual risk factors (insertion-deletion polymorphism of CCR5 and SNP G252A LTA) or in combination with other alleles or genotypes. The carriership of bi-or triallelic combinations was associated with MI more reliable than the carriership of any subsets: single alleles or allele pairs. The protective triallelic combination d*CCR5 + 252G*LTA + -174C*Il-6 was found to be the most significant (p = 0.0006, OR = 0.23, CI = 0.090–0.56). Separate analysis of genetic susceptibility to MI in men and women demonstrated sexual dimorphism for the CCR5 gene.

Proinflammatory cytokines interleukin-6 (IL-6), interferon-γ (IFNg) and tumor necrosis factor (TNF) play an important role in the pathogenesis of multiple sclerosis. Based on the published data concerning the effects of the SNPs G(−308)A of TNF, A(+874)T of IFNG, and G(−174)C of IL-6 on the production of these cytokines, we investigated the relation of these polymorphisms with multiple sclerosis. Linkage and association of alleles of these genes with multiple sclerosis were analyzed by transmission disequilibrium test. In a group of 104 nuclear families of Russian ethnicity, the TNF*(−308)A allele was more frequently transmitted from healthy heterozygous parents to affected children (p = 0.01). Linkage/association of IFNG and IL-6 alleles with multiple sclerosis was not detected. Thus, the data obtained indicate that TNF is involved in susceptibility to multiple sclerosis in Russians.

UNLABELLEDCRP level is a risk factor of development of ischemic heart disease (IHD) and acute myocardial infarction (MI) in healthy people, while in patients with cardiovascular diseases it is a marker of unfavorable prognosis. It has been shown in recent investigations that individual variations of plasma CRP levels to a great extent are genetically determined. These data constitute a basis for the study of associations of polymorphic variants of the CRP gene with risk of MI in healthy people as well as with unfavorable prognosis in IHD patients.MATERIAL AND METHODSWe included into the study 232 Russian patients aged 52.3 +/- 10.3 years, 175 men (50.1 +/- 10.6 years) and 57 women (55.2 +/- 10.1 years). Control group comprised 159 Russians without history of cardiovascular diseases and other serious severe concomitant diseases (age 60.5 +/- 14 years), 76 men (age 57.3 +/- 13.9 years ) and 83 women (age 63.1 +/- 14 years). CRP concentration was measured initially (at the moment of hospitalization), on days 3, at discharge, in 1 and 6 months, 1 year after onset of infarction. For genomic typing of C1444T polymorphism of CRP gene we used restriction fragment length analysis of products of polymerase chain reaction (PCR).RESULTSDistribution of genotypes of C1444T polymorphism of CRP gene: C/C 51.8%, C/T 35.8%, T/T 12.4% in patients with MI; C/C 55.2%, C/T 40.2%, T/T 4.6% in control group. We found significant difference (p = 0.006, relative risk [RR] 0.3, 95% confidence interval [CI] 0.15-0.74) in frequency of carriers of C1444 allele (sum of C/C and C/T genotypes), which was higher in control group. Correspondingly in the group of patients with MI T/T genotype was met significantly more frequently than in control (p = 0.006, RR 3.0, 95% CI 1.3-6.5), and can be looked upon as risk factor of MI. We found no relation between carriage of CRP alleles/genotypes of CRP and one year prognosis in patients with MI. Analysis of association of the C1444T polymorphism with CRP concentration revealed significant relationship between T/T genotype and higher CRP level.

Proinflammatory cytokines Interleukin-6 (IL-6), Interferon-gamma (IFNg) and Tumor necrosis factor (TNF) are known as participants of inflammation and play an important role in pathogenesis of multiple sclerosis (MS). Based on literature data about influence of SNPs G(-308)A of TNF gene, A(+874)T of IFNG gene and G(-174)C of IL-6 gene on production of these cytokines, we investigated association of these polymorphic sites with MS. Linkage and association of alleles of these genes with MS was analyzed by transmission disequilibrium test (TDT). In investigated group of 104 nuclear families of Russian ethnicity it was found that TNF* (-308)A allele transmitted from healthy heterozygous parents to affected children more frequently (p = 0.01). Linkage/association of IFNG and IL-6 alleles with MS was not revealed. Thus, data obtained indicate the participation of TNF gene in MS susceptibility in Russians.

Проведен анализ частот встречаемости аллелей и генотипов функционально значимых полиморфных участков генов, белковые продукты которых участвуют в развитии воспаления: генов провоспалительных цитокинов IL-6, LTA и TNF, гена антивоспалительного цитокина TGFB1 и гена рецептора хемокинов CCR5, у больных инфарктом миокарда этнических русских (199 человек) и в контрольной группе лиц той же этнической принадлежности (142 человека). Комплексный анализ с помощью алгоритма APSampler показал, что полиморфные варианты каждого гена – по отдельности (инсерционно-делеционный полиморфизм CCR5 и однонуклеотидный полиморфизм G252A LTA) или только в составе сочетаний с другими аллелями/генотипами – ассоциированы с инфарктом миокарда. Носительство сочетаний из двух или трех аллелей более значимо ассоциировано с инфарктом миокарда, чем носительство входящих в сочетания отдельных аллелей или пар аллелей. Наиболее значимым оказалось протективное триаллельное сочетание d*CCR5 + 252G*LTA + –174C*Il-6 (р = 0.0006, ОШ = 0.23, ДИ = 0.090–0.56). Анализ генетической восприимчивости мужчин и женщин к инфаркту миокарда выявил существование полового диморфизма по гену CCR5.

Carriage frequencies of alleles and genotypes of 10 functionally important single nucleotide polymorphisms that are located in genes FGA, FGB, APOE, LPL, ACE and CMA1 were analyzed in the ischemic stroke (IS) patients of Russian ethnic descent and in the control group of the same ethnic descent and of similar gender and age. Comparison between patients and control group revealed no significant differences in frequencies of individual alleles and genotypes for all the polymorphic loci studied. However, complex analysis of genetic predisposition using APSampler algorithm revealed carriage of allele (-491A) APOE as a predisposing factor for IS (p = 0.044, OR 3.8, 95% CI 1.0-15.1). Accordingly, carriage of genotype (-491T/T) APOE was associated with resistance to IS (p = 0.044, OR 0.26, 95% CI 0.07-1.0). The allele -249C FGB carriage addition to this genotype enhances its protective properties, p-value of the combination is 2-fold lower than that of the genotype (-491T/T) APOE (OR 0.17, 95% CI 0.04-0.8). Two more protective combinations were identified: biallelic (-427C) APOE + (1595G) LPL and triallelic (-491C) APOE + (1595G) LPL + (-1903G) CMAI (in both cases p = 0.0052, OR 0.18, 95% CI 0.05-0.66). Overall, involvement in formation of the risk of IS development in Russians was evidenced for alleles of four genes: APOE, FGB, LPL and CMA1, where APOE gene involvement was evidenced for alleles of two polymorphic loci, -491T and -427C. Linkage analysis suggested that involvement of these loci in insusceptibility to IS is mutually independent.

The frequencies of alleles and genotypes for ten functionally significant single-nucleotide polymorphisms were determined in the FGA, FGB, APOE, LPL, ACE , and CMA1 genes for Russian ischemic stroke (IS) patients and for a control group of Russians similar in gender and age distribution. The groups showed no significant differences in the frequencies of individual alleles or genotypes for any polymorphism studied. However, complex analysis of genetic susceptibility by the APSampler algorithm demonstrated that carriership of the APOE (−491A) allele predisposed to IS ( p = 0.044, OR 3.8, 95% CI 1.0–15.1). Correspondingly, the APOE (−491T/T) genotype was associated with resistance to IS ( p = 0.044, OR 0.26, 95% CI 0.07–1.0). The carriership of FGB (−249C) allele together with this genotype enhanced its protective potential, reducing the p value of the combination twofold (OR 0.17, 95% CI 0.04–0.8). Two more protective combinations were identified: biallelic APOE (−427C) + LPL (1595G) and triallelic APOE (−491C) + LPL (1595G) + CMA1 (−1903G). In both cases, p = 0.0052, OR 0.18, and 95% CI 0.05–0.66. Altogether, involvement in the formation of IS risk in Russians was evidenced for alleles of four genes: APOE, FGB, LPL , and CMA1 ; the APOE involvement was demonstrated for alleles of two polymorphic loci: −491T and −427C. Linkage analysis suggested that these loci were involved in IS resistance independently of each other.

For the first time an analysis of contribution of alleles of haemostasis genes, encoding fibrinogen alpha (FGA), fibrinogen beta (FGB) and tissue plasminogen activator (TPA), in genetic susceptibility to and clinical aspects of hemorrhagic stroke (HS) was performed in ethnic Yakuts. HS patients compared to controls demonstrated significant increase of frequency of FGA4266A/A genotype carriage: 26,9% vs 16,1% (p=0,04, OR=1,9, CI 1,0 - 3,8) and corresponding decrease of FGA4266G allele carriage: 73,1% vs 83,9% (p=0,04, OR =0,5, CI 0,3 - 1,0). We identified tri-allelic combination: FGB-249C; FGA4266G; TPA-7351C, which carriage is significantly differed in HS patients with different types of haemorrhage.

Проведен анализ частот встречаемости аллелей и генотипов 10 функционально значимых однонуклеотидных полиморфизмов в генах FGA, FGB, APOE, LPL, ACE и CMA1 в группе русских больных ишемическим инсультом и в контрольной группе лиц той же этнической принадлежности, сходной по полу и возрасту. Не обнаружено значимых различий в частотах отдельных аллелей и генотипов всех рассматриваемых полиморфных участков при сравнении группы больных и контрольной группы. Однако комплексный анализ генетической предрасположенности с использованием алгоритма APSampler показал, что носительство аллеля (–491A) APOE является фактором предрасположенности к ишемическому инсульту (р = 0.044, OШ 3.8, 95% ДИ 1.0–15.1). Соответственно, носительство генотипа (–491Т/T) APOE связано с устойчивостью к ишемическому инсульту (р = 0.044, ОШ 0.26, 95% ДИ 0.07–1.0). Добавление носительства аллеля –249C FGB к этому генотипу улучшает протективные свойства сочетания, снижая значение р в 2 раза (ОШ 0.17, 95% ДИ 0.04–0.8). Найдены еще два протективных сочетания: биаллельное (–427C) APOE + (1595G) LPL и триаллельное (–491C) APOE + (1595G) LPL + + (–1903G) CMA1 (в обоих случаях р = 0.0052, OШ 0.18, 95% ДИ 0.05–0.66). В целом, в формирование риска развития ишемического инсульта у русских вовлечены четыре гена – APOE, FGB, LPL и CMA1, причем у гена APOE – аллели двух полиморфных участков –491T и –427C. Результаты анализа сцепления позволяют предполагать, что эти участки вносят вклад в невосприимчивость к ишемическому инсульту независимо друг от друга.

Multiple sclerosis (MS) is a multifactorial disease of the central nervous system with pronounced hereditary predisposition. The purpose of the study was to test the assumption on the involvement of the apolipoprotein E gene (APOE) polymorphism in exon 4 in the development of MS in ethnic Russians. Samples independently collected in Moscow (106 MS cases and 189 control healthy volunteers), Sverdlovsk oblast (54 and 109, respectively), and the Republic of Bashkortostan (119 and 285, respectively) were examined. Genotypes for 2059C/T and 2197C/T polymorphisms of the APOE gene, which determine the amino acid substitutions C112R and R158C in apolipoprotein E, were analyzed by polymerase chain reaction followed by restriction analysis of amplificates. No statistically significant differences in genotype or allele frequencies were found between the control group and the group of MS cases. The APOE*4 allele is not associated with the risk of MS in ethnic Russians.

Arterial hypertension (AH) ranks among the most widespread cardiovascular diseases and is clinically and genetically heterogeneous. Except for rare monogenic forms, AH is polygenic, and an important role in AH predisposition belongs to genes of the renin-angiotensin-aldosterone system. Low renin activity in blood plasma is observed in 20–25% of AH cases (low-renin form of AH), while aldosterone production can be elevated (hyperaldosteronism, HA) or normal in these cases. Several polymorphisms of the genes coding for the renin-angiotensin-aldosterone system components were studied in patients with low-renin AH forms: primary HA, idiopathic HA, and AH with a normal aldosterone level. The chimeric CYP11B2/CYP11B1 gene, causing monogenic familial HA type 1, was absent from all HA cases studied. The patient groups were compared with respect to the allele and genotype frequency distributions of the polymorphisms of several genes (CYP11B2 (C-344T), REN (C-5434T, C-5312T, and A BglI G), AGT (Thr174Met), ACE (I/D), CMA (G-1903A), AT2R1 (A1166C)), and their combinations. Analysis of the carriership of the allele and genotype combinations implicated CYP11B2, REN, ACE, CMA, and AT2R1 in low-renin HA. The results revealed both similar and different features in the genetic nature of different low-renin AH forms. Investigation of the genetic predisposition to clinically heterogeneous forms of polygenic diseases by comparing patient groups, formed in accordance with peculiarities of the disease course, holds much promise for understanding their hereditary background.

We examined an association of the C-148T beta-fibrinogen gene polymorphism with fibrinogen level and platelet aggregation in patients with ischemic stroke and in people with vascular risk factors but no ischemic stroke. Among stroke patients, carriers of an -148T allele had significantly higher plasma fibrinogen level and platelet aggregation to norepinephrin compared to the C/C homozygotes. No significant differences in frequencies of a C-148 allele between stroke patients and controls were observed.

The multiple sclerosis is a complex disease of the central nervous system with the pronounced hereditary predisposition. The purpose of our research consisted in acknowledgement of the assumption on importance of apolipoprotein E gene (APOE) polymorphism in exon 4 in development of the multiple sclerosis in ethnic Russians. Research was lead on the samples independently collected in Moscow (106 patients and 189 persons of control group), Sverdlovsk area (54 and 109, accordingly) and republic Bashkortostan (119 and 285, accordingly). 2059C/T and 2197C/T polymorphisms of APOE gene, which determine aminoacid substitutions C112R and R158C in apolipoprotein E, were determined by polymerase chain reaction with the following restriction analysis of amplicons. There was not detected statistically significant distinctions on genotypes frequencies and alleles frequencies between control group and group of patients with multiple sclerosis. APOE*4 allele is not assosiated with risk of development of the multiple sclerosis at ethnic Russians.

We carried out comparison of distribution of alleles and genotypes of polymorphic loci of renin-angiotensin-aldosterone system genes: CYP11B2 (C-344T), AGT (Thr174Met) and REN (C-5434T, C-5312T, and A BglI G) and their combinations in two groups of patients with low renin forms of arterial hypertension (AH). Group 1 included 59 patients with low renin hyperaldosteronism (HA) at the background of glomerular zone adenoma and hyperplasia of adrenal cortex. Group 2 included 28 patients with low renin hypertensive disease characterized by normal level of aldosterone. Complex analysis of carriership of allele and genotype combinations evidence for the difference in genetic nature of two forms of low renin AH. Participation of CYP11B2 and REN and possibly AGT genes in development of low renin AH was convincingly shown.