[Five-year dynamics adrenergic reactivity of erythrocytes after radio-frequency sympathic denervation of renal arteries in patients with resistant arterial hypertension].

AIM: To study the initial state of adrenergic reactivity and the five-year dynamics of the beta-adrenergic reactivity index of erythrocyte membranes and the manifestation of the antihypertensive effect of the procedure for radiofrequency destruction of sympathetic structures of the renal artery in patients with resistant arterial hypertension. SUBJECTS AND METHODS: The analysis included 42 patients with resistant arterial hypertension (RH). The renal denervation (RD) procedure of the kidneys was performed by endovascular bilateral transcatheter radiofrequency ablation of the renal arteries. The study of 24-hour blood pressure monitoring (BPM) and the determination of ß-adrenoreactivity of erythrocytes (ß-ARM) by changes in the osmoresistance of erythrocyte membranes were performed initially, 1 week, 6 months, 1, 2, 3 and 5 years after RD. Patients retrospectively, at a follow-up period of 6 months after RD, were divided into responders (decrease in blood pressure by 10 or more mm Hg) and non-responders (decrease in blood pressure less than 10 mm Hg). RESULTS: 6 months after the RD, the number of responders was 28 people (66.7%), after 5 years - 31 people (73.8%). At the time of inclusion in the study, the median ß-ARM in the group of non-responders was not significantly higher than in the group of responders. After 6 months after the RD procedure, the ß-ARM indicator in the non-responder group was significantly lower than in the responder group (p = 0.043). With further follow-up in the group of responders, an increase in the median ß-ARM was noted, which reached significant differences relative to the baseline values in the group at follow-up periods of 1 year (p = 0.036) and 5 years (p = 0.004) after RD. The change in the ß-ARM indicator in the non-responder group was wavy in nature, the changes did not reach the significance criteria. CONCLUSION: Renal denervation in 73.8% of cases is accompanied by a stable antihypertensive response for 5 years of observation and an increase in ß-ARM, which may indicate the implementation of compensatory mechanisms in conditions of increasing activity of the sympathoadrenal system in response to a decrease in blood pressure.

[Methylation of Regulatory Regions of DNA Repair Genes in Carotid Atherosclerosis].

The status of DNA methylation in the human genome changes during the pathogenesis of common diseases and acts as a predictor of life expectancy. Therefore, it is of interest to investigate the methylation level of regulatory regions of genes responsible for general biological processes that are potentially significant for the development of age-associated diseases. Among them there are genes encoding proteins of DNA repair system, which are characterized by pleiotropic effects. Here, results of the targeted methylation analysis of two regions of the human genome (the promoter of the MLH1 gene and the enhancer near the ATM gene) in different tissues of patients with carotid atherosclerosis are present. Analysis of the methylation profiles of studied genes in various tissues of the same individuals demonstrated marked differences between leukocytes and tissues of the vascular wall. Differences in methylation levels between normal and atherosclerotic tissues of the carotid arteries were revealed only for two studied CpG sites (chr11:108089866 and chr11:108090020, GRCh37/hg19 assembly) in the ATM gene. Based on this, we can assume the involvement of ATM in the development of atherosclerosis. "Overload" of the studied regions with transcription factor binding sites (according to ReMapp2022 data) indicate that the tissue-specific nature of methylation of the regulatory regions of the MLH1 and ATM may be associated with expression levels of these genes in a particular tissue. It has been shown that inter-individual differences in the methylation levels of CpG sites are associated with sufficiently distant nucleotide substitutions.

Expression of Genes and Proteins of the Sarcoplasmic Reticulum Са2+-Transport Systems in Cardiomyocytes in Concomitant Coronary Heart Disease and Type 2 Diabetes Mellitus.

We compared the expression of Са2+-ATPase (SERCA2a), calsequestrin (CASQ2), ryanodine receptors (RyR2) proteins and their genes (ATP2A2, CASQ2, and RYR2) in coronary heart disease (CHD) patients with and without comorbid type 2 diabetes mellitus. All studies were performed on the right atrial appendages resected during coronary bypass surgeries. Expression of SERCA2a and RyR2 proteins and their ATP2A2 (p=0.046) and RYR2 genes in comorbid pathology was significantly (p=0.042) higher (by 1.2 and 2 times; p=0.025). The expression of CASQ2 protein and its gene did not differ significantly between the groups (p=0.82 and p=0.066, respectively). It was concluded that the expression of SERCA2a and RyR2 proteins and their genes (but not CASQ2 and its gene) is elevated in CHD associated with type 2 diabetes mellitus. Expression of the studied proteins correlated with the expression of their genes. Increased expression of CASQ2 protein and its gene can probably prevent imbalance of the Ca2+-transporting systems in cardiomyocytes and contractile dysfunction of the myocardium, even in CHD associated with type 2 diabetes mellitus.

Age-Related Features of the Viscosity of Plasma and Mitochondrial Membranes of Hepatocytes in Liver Cirrhosis.

The viscosity of plasma and mitochondrial membranes of hepatocytes was studied in young (3-month-old) and old (9-month-old) male Wistar rats. It was shown that viscosity of hepatocyte plasma and mitochondrial membranes in young rats under optimal vital functions in the area of protein-lipid membrane contacts was significantly lower than in old rats. No age-related differences in the viscosity of lipid-lipid membrane contacts and in the polarity of protein-lipid contacts and lipid layers were found. Liver cirrhosis induced by carbon tetrachloride and ethanol administration was associated with increased fluidity of the plasma and mitochondrial membranes of hepatocytes in rats of both age groups. The decrease in membrane viscosity in young rats occurred due to a decrease of the viscosity in the area of protein-lipid and lipid-lipid contacts, while in old rats in the area of protein-lipid contacts. Carbon tetrachloride and ethanol did not affect the polarity of lipid contacts and lipid layers.

Erythrocyte membranes beta-adrenoreactivity changes after renal denervation in patients with resistant hypertension, relationship with antihypertensive and cardioprotective intervention efficacy.

Aim      To study the functional condition of sympathoadrenal system as evaluated by beta-adrenoreactivity of erythrocyte membranes (beta-ARM) during two years following renal denervation (RD) in patients with resistant arterial hypertension (RAH) and to determine the relationship of this index with long-term antihypertensive and cardioprotective effectivity of this invasive treatment.Material and methods  The study included 48 patients (mean age, 57.2±8.7 years, 18 men) with RAH on a stable antihypertensive therapy. Averaged daily systolic and diastolic blood pressure (SBP and DBP) and levels of beta-ARM were determined at baseline and in 7 days and 2 years following RD. Measurement of beta-ARM was based on beta-adrenoblocker inhibition of erythrocyte hemolysis induced by exposure to hypo-osmotic environment. The beta-adrenoblocker binds to erythrocyte membrane beta-adrenoceptors to prevent the erythrocyte destruction. Increased values of beta-ARM reflect a decrease in the number of functionally active erythrocyte membrane beta-adrenoceptors associated with long-term sympathetic hyperactivity.Results For two years of follow-up, values of average daily BP decreased from 160.4±16.0 / 88.1±14.6 to 145.3±19.3 / 79.4±13.6 mm Hg. At 7 days, the number of beta-ARM had decreased in the group of RD responders (р=0.028) who at two years had decreased their BP by 10 mm Hg or more, while in the group of non-responders, the number of beta-ARM remained unchanged. At one week, beta-ARM values correlated with changes in SBP and DBP (r= -0.54; р<0.05) and with left ventricular myocardial mass (LVMM) (r= -0.36; р<0.05) at two years of follow-up whereas beta-ARM delta at one week was interrelated with the renin concentration in the long-term (r= -0.44; р<0.05). At two years, the content of beta-ARM was increased in both groups.Conclusion      The decrease in beta-ARM content at 7 days after RD shows the procedure efficacy and allows an expectation of clinically significant decreases in BP and LVMM in the long-term after the surgical treatment. At two years after the intervention, the content of beta-ARM increased, and the BP decrease was apparently due to some other mechanisms.

[Dynamics of adreneractivity after transfer of myocardial infarction: annual observation].

AIM: To study the change in the -adrenergic reactivity of red blood cell membranes in patients during the first year after acute myocardial infarction. MATERIALS AND METHODS: The study included 25 patients with acute myocardial infarction (AMI) who signed informed consent to participate in the study. The erythrocyte membrane -adrenoreactivity index (-ARM) was determined in venous blood samples upon admission to the intensive care unit, one day after admission, 6 and 12 months after the index MI was transferred using the BETA-ARM-AGAT reagent kit (Agat-Med, Russia). RESULTS: According to the results of dynamics assessment of -APM during the first day, patients included in the study were divided into 2 groups. Group 1 (n=14) included patients who had an increase in -APM in the first day, and group 2 (n=21) included patients in whom -ARM either did not change or decreased. At the time of admission to the hospital in the formed groups, there were no differences in the -APM index and clinical and anamnestic characteristics. A day after hospitalization, the value of -APM in group 1 significantly exceeded the same indicator in group 2 (p=0.02). At the periods of 6 and 12 months, the -APM indices in the groups did not differ. In the 2nd group of patients, the progression of chronic heart failure to one or more functional classes (NYHA) was significantly more often compared with the 1st group. CONCLUSION: The study showed that on the first day in patients with AMI, both an increase and a decrease in the activity of the sympathoadrenal system are possible with a further leveling of these differences over the next year. For a group of patients with decreased activity of sympathoadrenal system on the first day, a more unfavorable course of heart failure in the post-infarction period is characteristic.

Peculiarities of Cell Seeding on Electroformed Polycaprolactone Scaffolds Modified with Surface-Active Agents Triton X-100 and Polyvinylpyrrolidone.

We compared the capability of human fibroblasts to populate porous polycaprolactone (PCL) scaffolds modified during fabrication with surface-active agents Triton Ð¥-100 (type 1 scaffold) and polyvinylpyrrolidone (type 2 scaffold). The mean fiber diameter in both scaffolds was almost the same: 3.90±2.19 and 2.46±2.15 µ, respectively. Type 1 scaffold had higher surface density and hydrophilicity, when type 2 scaffold was 1.6 times thicker. The cells were seeded on the scaffolds by the dynamic seeding technique and then cultured in Petri dishes with nutrient medium in a humid atmosphere. During 3-day culturing, no cell release from the matrix was noted. DAPI staining proved the presence of cells in both scaffolds. However, in type 1 scaffold the cells populated the whole thickness, while in type 2 scaffold, the cells were present only in the superficial layer. These findings suggest that PCL scaffolds modified with Triton Ð¥-100 or polyvinylpyrrolidone are not cytotoxic, but the structure of the scaffold treated with Triton Ð¥-100 is more favorable for population with cells.

Peculiarities of the Functional State of Mitochondria of Peripheral Blood Leukocytes in Patients with Acute Myocardial Infarction.

We studied the rate of oxygen consumption by mitochondria isolated from peripheral blood leukocytes of patients with acute myocardial infarction and healthy volunteers. It was found that leukocyte mitochondria in patients with acute myocardial infarction were characterized by significantly lower rate of oxygen consumption and lower level of coupling of oxidation and phosphorylation processes in comparison with mitochondria from healthy volunteers.

[Comparative analysis of gene expression in vascular cells of patients with advanced atherosclerosis]. / Sravnitel'nyi analiz ékspressii genov v kletkakh sosudov u bol'nykh s klinicheski vyrazhennym aterosklerozom.

In this study we performed a comparative gene expression analysis of carotid arteries in the area of atherosclerotic plaques and healthy internal mammary arteries of patients with advanced atherosclerosis by using microarray HumanHT-12 BeadChip ("Illumina"). The most down-regulated genes were APOD, FABP4, CIDEC and FOSB, and up-regulated gene was SPP1 (|FC|>64; pFDR<0.05). The majority of differentially expressed genes were down-regulated in advanced atherosclerotic plaques. Unexpectedly, genes involved in immune and inflammatory responses were down-regulated in advanced atherosclerotic plaques to compare with the healthy arteries (arachidonic acid metabolism, cytokine-cytokine receptor interaction, NOD-like receptor signaling pathway, Jak-STAT signaling pathway, TNF signaling pathway). "Cellular response to metal ion" (metallothioneins) and "Extracellular matrix organization" were the most significant Gene ontology terms among the down- and up-regulated genes, respectively.

Peculiarities of Cell Seeding on Polylactic Acid-Based Scaffolds Fabricated Using Electrospinning and Solution Blow Spinning Technologies.

We studied the possibility of seeding bone marrow-derived stromal cells onto polylactic acid-based scaffolds fabricated by electrospinning and solution blow spinning technologies. The cells were applied to the scaffolds by dynamic seeding and scaffolds were then cultured in Petri dishes in culture medium for 3 days. Cell migration to the Petri dish surface was noted only for scaffolds fabricated by electrospinning technology, but DAPI staining confirmed the presence of cells in both scaffolds. The mean number of cells in scaffolds fabricated by electrospinning and solution blow spinning was 56±9 and 81±6, respectively. The scaffold fabricated by solution blow spinning was more effectively (p<0.05) colonized by cells due to its more optimal spatial structure.

[Association of ITGB3, P2RY12, and CYP2C19 gene polymorphisms with platelet functional activity in patients with coronary heart disease during dual antiplatelet therapy]. / Assotsiatsiia polimorfizmov genov ITGB3, P2RY12, CYP2C19 s funktsional'noi aktivnost'iu trombotsitov u patsientov s ishemicheskoi bolezn'iu serdtsa na fone dvukhkomponentnoi antiagregantnoi terapii.

AIM: To assess the association of CYP2C19 G681A, P2RY12 H1/H2, and ITGB3 T1565C polymorphisms with the extent of platelet aggregation in patients with coronary heart disease (CHD) during antiplatelet therapy. SUBJECTS AND METHODS: 166 male patients with CHD, living in the Western Siberian Region, were examined. All the patients underwent a test for platelet aggregation induced by ADP (2.5 and 5.0 µm) and epinephrine (0.2 µm). Genotyping was performed using an allele-specific polymerase chain reaction technique. RESULTS: The polymorphic variants of the P2RY12 and ITGB3 genes were ascertained to have no impact on the extent of platelet aggregation in patients receiving clopidogrel and acetylsalicylic acid. An association was found between CYP2C19 681A allele carriage and the increased extent of platelet aggregation induced by ADP. CONCLUSION: The carriage of the cytochrome P450 CYP2C19 681A allele rather than platelet receptor gene polymorphisms determines a risk for clopidogrel resistance in patients with CHD.

[Possibility of evaluating the effectiveness of renal artery sympathetic denervation in resistant hypertension early after radiofrequency ablation]. / Vozmozhnost' otsenki effektivnosti simpaticheskoi denervatsii pochechnykh arterii pri rezistentnoi arterial'noi gipertonii v rannie sroki posle provedeniya radiochastotnoi ablyatsii.

AIM: to investigate time course of changes in the adrenoreactivity (AR) of erythrocyte membranes (EM) after radiofrequency ablation of the synaptic nerves of the renal arteries (RA) in patients with resistant hypertension (RH) and to assess whether this indicator can be used for the early evaluation of the efficiency of an invasive intervention into the RA. SUBJECTS AND METHODS: 24 patients with RH, who received full-dose antihypertensive therapy with at least three drugs, including a diuretic, were examined. Renal sympathetic denervation (RSD) was carried out by endovascular radiofrequency ablation (RFA) of the RA. 24-hour blood pressure (BP) monitoring and determination of the ß-adrenoreactivity (ß-AR) of EM were performed, by taking into account the change in erythrocyte osmoresistence at baseline and 1 and 24 weeks after RFA. The therapy was not changed during the observation. RESULTS: The patients included in the study were divided into 2 groups. One week following RSD, 15 patients of Group 1 were noted to have a decrease in the ß-AR of EM by 10 conditional units or more; average daily systolic BP (SBP) and diastolic BP (DBP) reduced by 8.3 and 2.8 mm Hg, respectively. In 9 patients of Group 2, the ß-AR of EM was unchanged in this observation period or increased compared with baseline. In this group, the decrease in average daily SBP and DBP was noted to be less pronounced than that in Group 1 (by 1.4 and 1.5 mm Hg, respectively). At 24 weeks after RSD, Group 1 was seen to have an effective daily decrease in average daily SBP and DBP by 25.6 and 14.3 mm Hg, respectively (p=0.01 and 0.05). The average value of the ß-AR of EM significantly declined compared with baseline. In Group 2, no statistically significant changes were reported; SPB and DBP lowered by 7.0 and 3.0 mm Hg, respectively. There was a significant decrease in the ß-AR of EM compared with that at week 1. CONCLUSION: The decline in the ß-AR of EM within the first week after RFA is suggestive of the decreased activity of the sympathoadrenal system and may be used as an early efficiency index of RSD after the procedure.

[Association of I/D and -786 Polymorphisms of ACE and NOS3 Genes With Features of the Course of Ischemic Heart Disease and Diabetes Mellitus Type 2].

AIM: to study relationship of ACE insertion-deletion (I/D) polymorphism and NOS3 T-786C polymorphism with characteristics of the course of ischemic heart disease (IHD) at the background of diabetes mellitus. MATERIALS AND METHODS: Were examined 114 patients with IHD, 29.8% of patients had type 2 diabetes mellitus. ACE and NOS3 polymorphisms were determined by allele-specific polymerase chain reaction with primers by "Lytech". RESULTS: Patients with combined pathology belonged to older age group, had increased frequency of obesity and predominance of functional class II chronic heart failure. In this group we detected association of D allele of the ACE gene with higher frequency of dyslipidemia and obesity. Among patients with IHD without diabetes we observed associations of ACE I/D and NOS3 T-786C polymorphisms (close and moderate, respectively) with severity of effort angina. We also found that frequency of dyslipidemia among carriers of II and TT genotypes was lower than among carriers of other genotypes. CONCLUSION: Presence of type 2 diabetes as background pathology leads to a change of character of association of ACE I/D and NOS3 T-786C polymorphisms with clinical characteristics of patients with IHD.

[Manifestations of polymorphism of ß1-adrenoreceptors in patients with newly diagnosed cardiac rhythm disorders].

Individual peculiarities of the receptor apparatus of cardiomyocytes may determine pathological features of heart activity and susceptibility to pharmaceuticals. The possible role of beta-adrenoreceptor polymorphism in the development of cardiac rhythm disturbances is assessed by PCR. Special attention is given to A145G polymorphism of the ADRB1 gene in 127 patients with primary cardiac rhythm disorders. It was shown that AJ45G polymorphism (Ser49Gly) at DNA sites encoding for the amino acid sequence of beta-1 adrenoreceptors can influence the development of sex-specific cardiac rhythm disorders.

[Resistance to clopidogrel and polymorphism of P2RY12 and GPIIIA genes in patients with chronic coronary heart disease].

We studied the association between clopidogrel resistance, H1/NH2 polymorphism of the P2RY12 gene and T156C polymorphism of the GpIIIa gene in residents of Western Siberia suffering chronic CHD. It was shown that the occurrence of H1 and H2 haplotypes of the P2RY12 gene and 1565T and 1565C alleles of the GpIIIa gene was similar to that reported for European populations. Patients showing variable platelet response to the inhibitory action of clopidogrel were not significantly different in terms of P2RY12 and GpIIIa genotype distribution. To conclude, the study revealed no association between the risk of clopidogrel resistance and the presence of polymorphic variants of platelet receptor genes P2RY12 and GpIIIa.

[D allele of the angiotensin converting enzyme gene is a putative risk factor of restenosis after coronary stenting in patients with coronary heart disease].

We undertook a comparative analysis of allele frequency distribution and I/D polymorphism of the angiotensin converting enzyme (ACE) gene in patients with coronary heart disease (CHD) undergoing coronary stenting with and without signs of restenosis. There were no significant difference between the groups in the genotype frequency distribution of I/D polymorphism of the CAE gene (p = 0.061). The occurrence of D allele in the patients with restenosis was higher than that of I allele (chi-square test 4.117, p = 0.042). Relative risk for the carriers of D and I alleles was 0.35 and 2.83 respectively. It is concluded that the presence of D allele of I/D polymorphism of the CAE gene in patients with chronic CHD may be a risk factor of restenosis of stented coronary arteries.