Expression of TLR2, IL-1ß, and IL-10 Genes as a Possible Factor of Successful or Pathological Aging in Nonagenarians.

The expression of the gene of pattern recognition receptor TLR2, proinflammatory cytokine IL-1ß, and anti-inflammatory cytokine IL-10 was analyzed in the peripheral blood of nonagenarians (n=219; mean age 92.1 years, 77 men and 142 women) in comparison with healthy young donors (n=24; mean age 22.5 years, 16 women and 8 men). Nonagenarians were interviewed, medical records were analyzed, and a comprehensive geriatric assessment was performed according to the Clinical Guidelines on Frailty. The level of gene expression was determined by real-time PCR. The participation of inflammatory mechanisms in the immunosenescence was revealed. It was shown that increased expression of IL1B and TLR2 genes is associated with the development of frailty in nonagenarians and can be a factor of pathological aging. Increased expression of IL10 gene can be considered as a factor of successful aging in nonagenarians.

[Features of innate immunity indicators in patients with recurrent exudative otitis media]. / Osobennosti pokazatelei vrozhdennogo immuniteta u patsientov s retsidiviruyushchim ekssudativnym srednim otitom.

The article presents the features of the indicators of the innate immune response (TLR4, IL1B, TGFB, HBD1, and HBD2) in the exudate of the tympanic cavity in patients with recurrent exudative otitis media (EOM) with normal patency of the auditory tube and its dysfunction. The results of the study demonstrate changes in the indices of the innate immune response characteristic of the inflammatory process in patients with recurrent EOM against the background of dysfunction of the auditory tube in comparison with the group where it is absent. The data obtained can be used to clarify the pathogenesis of otitis media with dysfunction of the auditory tube, to develop new methods of diagnosis, prevention and therapy.

[Hypertrophy of palatine tonsils - possible treatment approaches]. / Gipertrofiya nebnykh mindalin ­ vozmozhnye podkhody v lechenii.

OBJECTIVE: To evaluate the effectiveness of different approaches to the treatment of patients with hypertrophy of the palatine tonsils (HPT). MATERIAL AND METHODS: 90 children with HPT of II-III degree and 20 healthy children (group 1) aged from 3 to 7 years were included in the study. Children with HPT were divided into three groups: group 2 - children who underwent tonsillotomy (n=30), group 3 - children who underwent conservative treatment, including only topical use of Polyoxidonium (n=30), group 4 - children who underwent complex treatment (tonsillotomy with subsequent local use of Polyoxidonium), (n=30). The severity of nasal breathing disturbances on a visual analogue scale, the average number of acute respiratory viral infections 6 months before and after treatment, the level of gene expression of antimicrobial peptides before and after treatment, the degree of hypertrophy of the palatine tonsils and spleen with an assessment of the echostructure and determination of the spleen mass coefficient using Ultrasound were evaluated in the study. RESULTS: In children receiving only Polyoxidonium, there was a decrease in the severity of nasal breathing disorders, a decrease in the frequency of acute respiratory viral infections, an increase in the expression of antimicrobial peptide genes compared to the initial level of these indicators. Comprehensive treatment of children with HPT (group 4) showed a significant decrease in the severity of nasal breathing disorders, a decrease in the average number of acute respiratory infections within 6 months, an increase in the expression of antimicrobial peptide genes compared to children who underwent only tonsillotomy (group 2) or only conservative therapy (group 3). CONCLUSION: It is proved that the use of Polyoxidonium in the complex treatment of HPT is clinically effective and safe.

Effect of Natural Cytokine Complex on Metabolism of Smooth Muscle Cells in Myocardial Arteries under Normal Conditions and during Hemodynamic Overload.

Histoenzymological methods were employed to examine the effects of systemically administered natural cytokine complex including IL-1, IL-2, IL-6, TNFα, MIF, and TGFß on metabolism of smooth muscle cells in intramural myocardial arteries under physiological conditions and during acute hemodynamic overload of the heart. Natural cytokine complex markedly inhibited metabolism of vascular smooth muscle cells under control conditions and during acute experimental aortal stenosis. In vascular smooth muscle cells, deceleration of tricarboxylic acid cycle, redistribution of the fluxes in glycolytic cascade and its inhibition, down-regulation of oxidation of free fatty acids and their metabolites, and inhibition of the shuttle systems and biosynthetic processes were observed. Inhibition of metabolism in the vascular wall of myocardial arteries correlated with a decrease in their tone and could be partially determined by a decrease in contractile activity of smooth muscle cells. These findings do not exclude the involvement of other factors and mechanisms in down-regulation of metabolism in vascular myocytes in response to increased cytokines levels of in the blood, including their direct effect on biochemical processes in cells.

Effects of Natural Cytokine Complex on the Myocardial Blood Flow in Normal and under Conditions of Increased Hemodynamic Load.

The effects of a natural complex of cytokines IL-1, IL-2, IL-6, TNF, MIF, and GTFß on myocardial blood flow were studied under control conditions and during acute experimental aortal stenosis. Systemic administration of the cytokine complex under control conditions led to moderate impairment of the blood flow in the myocardium associated with plethora and perivascular edema. The number of functioning vessels in the myocardium significantly increased under these conditions, which reflected enhancement of the coronary blood flow. The comparison of the myocardial blood flow under conditions of acute heart overload alone and in combination with systemic administration of the cytokine complex revealed similar changes. In both cases, moderate plethora in all compartments of the vascular network, moderate perivascular edema, and moderate blood stasis in the myocardial capillaries were seen. The only difference the increase in the density of functioning capillaries that was significantly more pronounced after cytokine administration. These data indicate that the increase in the blood cytokine level induced dilatation of myocardial vessels and intensification of blood flow in normal and under conditions of acute hemodynamic heart overload. Against the background of pronounced vasodilatation, the dyscirculatory changes in the myocardium were moderate. It was assumed that the increase in the duration or frequency of hypercytokinemia episodes can induce more severe blood flow disturbances in the myocardium.

Combined Effect of TLR2 Ligands on ROS Production by Mouse Peritoneal Macrophages.

TLR2-mediated ROS production by mouse peritoneal macrophages was studied by luminoldependent chemiluminescence under conditions of cell stimulation with zymosan (TLR2/6 ligand) and peptidoglycan (TLR2/1 ligand). ROS production by macrophages stimulated with zymosan and peptidoglycan simultaneously depended on the ratio of ligand concentrations. Three effects were revealed: additivity of the stimulating effects of the ligands used, competitive ligand binding, and effect of macrophage priming with peptidoglycan during cell stimulation with zymosan. The mechanisms of these effects are discussed.

Effect of Natural Cytokine Complex on the Structure and Metabolism of the Cardiac Conduction System in the Myocardium under Normally and Increased Hemodynamic Load.

Effect of natural complex of cytokines with activity of IL-1, IL-2, IL-6, TNF, MIF, and GTFß on the structure and metabolism of conduction cardiomyocytes was assessed in the control and under acute experimental aortic stenosis. After systemic administration of the cytokine complex in the control, structural abnormalities were revealed in a relatively low number of conduction cardiomyocytes; their relative number increased in the left ventricle and interventricular septum. When the complex was administered against the background of aortic stenosis, morphological changes in the conduction system were seen in a significant number of cells with their plasma imbibition, especially in the left ventricle and interventricular septum. Systemic administration of the natural cytokine complex inhibited the major metabolic processes in the conduction system, both in the control and under conditions of sharply increased hemodynamic load. In conduction cardiomyocytes, deceleration of glycolysis and citric acid cycle, inhibition of oxidation of free fatty acids and their metabolites, and suppression of shuttle mechanisms and biosynthetic reactions were observed. Increased blood levels of cytokines, primarily of the proinflammatory ones, can cause structural and metabolic disturbances in the cardiac conduction system and promote the development of arrhythmias, especially in case of sharply increased hemodynamic load.

The role of regulatory RNAs (miRNAs) in asthma.

INTRODUCTION: Recently, a great deal of attention has been paid to the investigation of regulatory functions of microRNA. Currently, many different mechanisms involved in the pathogenesis of asthma are known, but the whole picture of pathogenesis has not yet been studied. CONCLUSIONS: MicroRNAs play an important role in the regulation of many cellular processes. Undoubtedly, these regulatory molecules are involved in the pathogenesis of asthma, and therefore can be potential targets for treatment.

[Study of cytokine profile in men with hypertension.]

Gender differences in the risk factors' influence on the development of cardiovascular diseases determine the study of the pathogenetic causes of these features. Inflammatory is one of the mechanisms for the participation of risk factors in the development of hypertension. The aim of the study was to investigate the relationship between inflammatory biomarkers with hypertension and cardiovascular risk factors in men. The study included 60 men with hypertension who were divided into groups depending on the risk of cardiovascular events: 30 men with low risk and 30 men with high risk. 86 hypertensive patients with risk factors such as obesity, impaired glucose metabolism, impaired lipid metabolism also were examined. Biochemical parameters and a wide range of cytokines: IL-4, IL - 8, IL-10, IL-12, IL-17, IL-18, TGF-ß, TNF-α, IFN-γ were estimated. The obtained results were compared with echocardiographic (EchoCG), ambulatory blood pressure monitoring (ABPM), duplex scanning of the carotid arteries. A direct correlation between the risk factors and target organs' damage - left ventricular hypertrophy, increased intima-media complex thickness, was revealed. The dependence of cytokine profile from risk of cardiovascular events in men was also revealed. We studied the contribution of individual risk factors in men in the development of hypertension and the severity of inflammation. More significant changes in the cytokine profile observed in patients with concomitant hyperglycemia, obesity and dyslipidemia. A correlation between cytokine profile and duration of hypertension also revealed. Correlation analysis revealed the relationship between ABPM parameters and levels of interleukins, HS-CRP, and fibrinogen. Biomarkers of inflammation associated with the severity of visceral obesity, with a BMI. Analysis of results also showed the relationship between hyperinsulinemia or insulin resistance and inflammatory biomarkers. The relationship of inflammation with lipid metabolism in hypertensive men was more significant than with the other studied risk factors.

[Study of heat shock protein gene 70 (HSP70) expression and its polymorphous marker (+ 1267A> G) in women under exposure to a long-term stress.]

There was studied the HSP70 gene and its polymorphic marker + 1267A> G (rs754888705) in the HSP70 gene expression in32 women under long-term stress, which was caused by inpatient treatment of their oncologic child. A control group included 25 women without long-term stress factors. There was studied a correlation between the expression level of the HSP70 gene in women and their high blood pressure (BP) episodes. The average duration of stress in the group of children's mothers was 7.3 (2.5-11.5) months. Anxiety level according to HADS in the main group was significantly increased and amounted to 8.7 (7-10) points, in the control group 5 (2-7) points. Depression level in the main group was significantly higher and amounted to 7.7 (7-9) points, in the control group 3.3 (1-5) points. Comparison of the HSP70 gene expression level in blood of young women of the main study group with expression level of this gene in the control group showed a statistically significant predominance of HSP70 gene expression in the main group. The episodes of high BP in women who were under stress conditions did not influence the expression level of the HSP70 gene. Analysis of the alleles of the polymorphic marker frequency in the gene HSP70-2 1267A> G (rs754888705) in the main and control groups showed a statistically significant predominance of allele A in the group of mothers under stress, and predominance of the allele G in the control group. Comparative analysis of the polymorphic marker genotype frequencies in the gene HSP70-2 1267A> G (rs754888705) showed that the AA genotype is significantly more frequent in mothers with episodes of BP increase compared to women without it. An increased level of the HSP70 gene expression in peripheral blood leukocytes in mothers with a life-threatening disease of their child indicates the damaging effect of long-lasting psychoemotional stress at the cellular level and activation of the protective reaction mediated by HSP70. Studying of the genotype AA characteristics of the polymorphic marker A1267G (rs754888705) of the HSP70-2 gene in women under long-lasting stress will let us evaluate the role of genetic factors in hypertensive reactions development.

Effect of the Natural Cytokine Complex on the Structure and Metabolism of Contractile Myocardium Normally and under Increased Hemodynamic Load.

Effect of natural complex of cytokines with activity of IL-1, IL-2, IL-6, TNF, MIF, GTFß on the structure and metabolism of contractile ventricular cardiomyocytes was assessed in the control and under conditions of acute experimental aortic stenosis. Systemic administration of the complex in the control had no significant effect on myocardial morphology with low number of damaged cardiomyocytes and low degree of structural damage. Administration of the cytokine complex against the background of aortic stenosis did not exert any additional alterative effect on cardiomyocytes, structural damage of contractual nature was moderate. Systemic administration of the natural cytokine complex had a pronounced inhibitory effect on metabolic processes in the myocardium of both ventricles both in the control and against the background of increased hemodynamic load. In cardiomyocytes, glycolysis and citric acid cycle were slowed down, oxidation of free fatty acids and their metabolic products was inhibited as well as shuttle mechanisms and biosynthetic reactions. Inhibition of energy-producing processes is the cause of the lack of the contractile function energy supply and can worsen the course of cardiovascular diseases and increase the risk of their complications in conditions, accompanied by increased blood cytokine level.

[The expression of TLR4 and HBD3 in epithelial cells of oral mucosa by surgical treatment of periodontitis]. / Ekspressiya genov TLR4 i HBD-3 v epitelial'nykh kletkakh slizistoi obolochki pri khirurgicheskom lechenii parodontita.

The expression of TLR4 and HBD3 in epithelial cells of the oral cavity was studied using polymerase chain reaction in real-time in patients with inflammatory and destructive periodontal lesions before and after surgery using osteoplastic material. Analysis of TLR4 and HBD3 expression in epithelial cells demonstrates the key role of the innate immune factors in the pathogenesis of inflammatory and destructive periodontal lesions. TLR4 gene expression was increased 1.5-7.0 times and HBD3 1.5-5.0 times compared to healthy individuals. Surgical treatment resulted in an effective rehabilitation and normalization of innate immunity.

[ROLE OF INNATE IMMUNITY FACTORS IN PERIODONTITIS PATHOGENESIS].

Chronic generalized periodontitis (CGP) is a disease of periodontium tissues supporting tooth induced by bacteria, that is characterized by the presence of processes of inflammation with destruction of bone tissue. The knowledge of molecular mechanisms of CGP pathogenesis facilitates creation of the most effective methods of therapy of this disease. Bacterial infection is a primary factor in periodontitis etiology, however is not sufficient for its start and subsequent development. It is known, that bacterial factors induce alocal inflammationreaction and.activate the system of innate immunity through activation of Toll-like receptors (TLR), located on the surface of resident cells and leukocytes. Activation of these cells results in production of pro-inflammatory cytokines and recruitment of phagocytes and lymphocytes into the inflammation zone. In review we examined the known data regarding factors of immune protection of periodontium including cell populations and cytokines, as well as mechanisms of tissue destruction, that support the tooth. Perspectives of therapy are also discussed

[ASSOCIATION OF POLYMORPHIC MARKERS IN GENES OF INNATE IMMUNITY IN PATIENTS WITH PERIODONTITIS AND INFLAMMATORY DISEASES OF UPPER RESPIRATORY TRACT].

AIM: Search of association of polymorphisms in DEFB1, IL-10, TNF-α and TLR2 genes with development of chronic generalized periodontitis in representatives of Ural region (Caucasian race). MATERIALS AND METHODS: 142 patients, that were split into 3 groups, took part in the study: a group of patients with periodontitis, a group with frequent inflammatory disease of upper respiratory tract and a comparison group--healthy donors. A study of polymorphic markers was carried out: DEFB1 (-44G/C), DEFB1 (-20A/G), IL-10 (-1082.A/G), TNF-α (-308 G/A), Arg753Gln and Arg677Trp using PCR in real time mode. RESULTS: Association of infectious pathology ofupper respiratory tract and development of periodontitis diseases with markers in DEFB1 (-44G/C) and Arg753Gln and Arg677Trp genes was determined. Significant differences in distribution of genotypes and alleles of genes IL-10 and TNF-α in the group of patients with periodontitis and comparison group were not detected. CONCLUSION: DEFB1 (-44G/C) polymorphism can be examined as a marker of periodontitis development risk.

In vitro studies of the antiherpetic effect of photodynamic therapy.

The number of viral infection cases in the Department of Gynecology and Obstetrics has tended to increase over last few years. Viruses form herpesvirus and cytomegalovirus families are associated with an increased risk for recurrent pregnancy loss. Photodynamic therapy (PDT) is a promising new approach to treat viral infections in which viral particles are inactivated. It exhibits great therapeutic potential, particularly among this group of patients. This study examined the use of PDT to treat herpesvirus infection (HVI) using an in vitro model. In this study, we used the Vero сell lineage as a suitable model of HVI, strains of HSV-1 (strain VR-3) and HSV-2 (strain MS) obtained from The National Virus Collection (London, UK), the photosensitizer Fotoditazine (Veta-Grand, Russia), an AFS physiotherapeutic device (Polironic Corporation, Russia). Laser light irradiation and the photosensitizer had different cytotoxic effects on the Vero cell cultures depending on the doses used. The optimal laser light and photosensitizer doses were determined. PDT had an antiviral effect on an in vitro model of HVI in cell culture. PDT has been shown to be effective treatment for HVI in vitro, leading to a reliable decrease of viral titer.

Association of Polymorphisms in Innate Immunity Genes TLR9 and DEFB1 with Human Longevity.

The distribution of polymorphic variants of innate immunity genes TLR9 (+2848G>A) and DEFB1(-20G>A; -44C>G; -52G>A) was evaluated in long-living individuals. No significant differences were found in the distribution of genotypes and alleles of the TLR9 gene. The following features were revealed: increase in the frequency of AA and GG genotypes and decrease in the frequency of the AG genotype of the DEFB1(-20G>A) gene; increase in the frequency of the CC genotype and C allele and decrease in the frequency of CG and GG genotypes and G allele of the DEFB1(44C>G) gene; and increase in the frequency of AA and AG genotypes and A allele and decrease in the frequency of the GG genotypes and G allele of the DEFB1(-52G>A) gene. Genotypes and alleles of the DEFB1 gene found in long-living individuals can be considered as the factors that increase the probability of longevity and favorable course of age-related diseases.

[STUDY OF THE ROLE OF INNATE IMMUNITY FACTORS (TLR2, HBD-2, TNF-α, TGF-ß) IN PERIODONTITIS PATHOGENESIS].

AIM: Study of the role of innate immunity factors, namely, expression of TLR2 and HBD-2 genes, as well as TNF-α arid TGF-ß in pathogenesis of periodontium tissue inflammation. MATERIALS AND METHODS: 59 individuals with periodontitis illnesses were included into the study; 15 healthy donors represented the comparison group. Study of the TLR2 and HBD-2 gene expression levels was carried out by rRT-PCR, cytokine production evaluation--by EIA. RESULTS: The results of the study have shown the presence of TLR-mediated disbalance in the innate immunity system in the periodontium tissue during chronic generalized periodontitis. TLR2 gene hyperexpression was accompanied by the reduction of expression of anti-microbial peptide HBD-2, as well as an increase of production of TNF-αand TGF-P by epithelial cells of periodontium mucosa. Conclusion.-The study carried out has shown that disturbance of molecular mechanisms of innate immunity system has an important place in pathogenesis of periodontitis. -

Expression of TLR9 and BD-2 protein genes in corneal cells of mice of different strains with herpetic keratitis.

The dynamics of gene expression of two proteins, TLR9 (one of the key receptors recognizing CpG repeats of herpes virus DNA) and ß-defensin 2 (antibacterial peptide), was studied on the model of herpetic keratitis in C57Bl/6 and BALB/c mice. New data on differences in TLR9 gene expression in mice of the two strains infected with the virus were obtained. Reduced TLR9 gene expression in the cornea of C57Bl/6 mice was associated with their high sensitivity to infection caused by herpes simplex 1 virus.

Natural complex of cytokines is a potent stimulant to posttraumatic regeneration in rabbit cornea.

We elaborated an original technique based on local application of natural complex of cytokines (NCC) secreted by autologous peripheral leukocytes. In this study, we evaluated NCC influence on the healing of penetrating corneal wounds. NCC was derived from supernatants of PHA-stimulated rabbit peripheral leukocytes. Biological tests revealed the presence of IL-1, TNF, IL-6, MIF, and LIF in the complex. Chinchilla rabbits with standard penetrating corneal wounds received daily NCC instillation. The controls were instilled with cultural media 199 with antibiotics. At 24 hours, 3, 7, 14 and 30 days post-wounding, the rabbits were euthanized. We performed morphometry of corneal cross-sections stained with hematoxylin and eosin. In NCC-treated animals, we observed more vigorous migration and activation of neutrophils and macrophages followed by augmented resorption of fibrin. The later post-injury period (14-30 days) was marked with complete healing of the endothelial defect (in the controls, the mass of proliferating cells projected into the anterior eye chamber) and mature scar tissue with a higher content of fibrous component. NCC-treated eye scars were 1.6-fold thinner than the controls'. Local application of NCC promotes effective healing of posttraumatic cornea. It regulates all stages of regeneration and prevents rude scarring.