[Study of the structure and microflora of urethral tissues in urethral pain syndrome].

Urethral pain syndrome (UPS) is characterized by the occurrence of persistent or recurrent pain in the urethra in the absence of a confirmed infection and other obvious local pathological changes. The study of its pathogenetic aspects is important first of all for understanding the causes of the disease, to prescribe effective treatment, specific recommendations for the prevention and treatment of this disease are also absent. This paper presents the advanced experience of our research group on the study of the urethral state by the in vivo cross-polarization optical coherence tomography (CP OCT) method, and also the results of the microbiota analysis in the urethral tissues. The purpose of the study is to search for the risk factors for UPS and the character of changes in the urethral tissues, using the data of: 1) concomitant pathology, 2) structural changes in the urethral wall in UPS in comparison with chronic cystitis of bacterial etiology 3) studying the microbiota of urethral tissues. MATERIALS AND METHODS: The condition of the urethra was studied in 109 patients: 55 of them with UPS (group "US"), without clinical manifestations of inflammation; 41 - with chronic inflammation of the lower urinary tract of various origins (group "Inf"); in 14 patients with stones of the upper urinary tract without pyelonephritis, the urethra was taken as the norm (group "N"). All performed a clinical minimum of studies, also cystoscopy with the study of the bladder triangle, the neck of the bladder and the urethra by the method of in vivo tissue imaging - CP OCT. The device "OCT-1300U" with wavelength of 1300 nm is used. To determine the possible role of UPS disease background, the analysis of concomitant pathology preceding the development of UPS was performed. To analyze the relationship of changes in the urethral tissues with the composition of its microbiota, a PCR study of biopsies from the proximal segment of the urethra was performed in 13 patients with UPS. RESULTS: Qualitative comparison of the thickness and character of the OCT signal of the urethral wall layers observed using CP OCT in the studied groups of patients allowed us to establish that the state of the epithelium and connective tissue structures of the mucous membrane in patients with UPS is not the norm, changes are similar to those in chronic inflammation. Changes in the character of the OCT signal were recorded in all parts of the urethra, but in the middle third they are most pronounced and most critical. In UPS, there is a brightly pronounced reorganization of the connective tissue stroma components. Pronounced fibrosis of subepithelial structures (increased signal brightness in the cross-channel compared to the norm) with their thickening was recorded in 48.2% of cases, and thinning/lack of visualization of the epithelial layer was detected in 20.5%, and in chronic inflammation 55.5% and 40.6% of cases, respectively. According to the results of PCR, only one patient had significant total bacterial contamination of the biopsy (TB=104.7). In all other cases, the total bacterial mass of the biopsies was at the level of negative control. CONCLUSIONS: In patients with UPS, the presence of several concomitant, often chronic, diseases was revealed, which may be a premorbid background and one of the risk factors for the occurrence and maintenance of UPS. Pilot PCR studies of biopsies from the proximal segment of the urethra indicate that low values of bacterial contamination in the majority of patients with UPS do not exclude the possible role of bacteria in the development of the disease in some patients. The CP OCT method used in this study is currently the only one in vivo method of visualization of the urethral mucosa, which provides real-time images of structural changes in the epithelial (atrophy or hyperplasia) and connective tissue (active or latent inflammation with cellular infiltration or fibrosis) layers of the urethra, allowing better understanding of the pathogenesis of the disease and monitoring of therapy.

[Evaluation of the composition of the microbiota of the urethra in men with sexually transmitted infections].

OBJECTIVE: to conduct a comparative study of the composition of the microbiota of the urethra in men with sexually transmitted infections (STIs), and healthy men. MATERIAL AND METHODS: The study included 103 men aged 18 to 45 years: 42 men with urethritis caused by STIs and 61 clinically healthy men. Identification of pathogenic and conditionally pathogenic microorganisms in scrapings from the urethra was performed by PCR in real time (test system Androflor (DNA-Technology, Moscow). RESULTS: In the analysis of the total bacterial mass, it was found that the bacterial contamination of the urethral biotope in patients with STI was significantly higher than in the group of healthy men (5.8 Lg10 and 4.7 Lg10, respectively), with the highest level of bacterial contamination was detected in patients infected with N. gonorrhoeae (6.4 Lg10). Patients with STIs had significantly lower levels of relative Staphylococcus spp., Streptococcus spp., Corynebacterium spp. and their amounts in General compared to clinically healthy men: according to ROC analysis, the best diagnostic indicator (0.93+/-0.04, p<0.001), distinguishing a group of healthy individuals from patients with STI, was the amount of Staphylococcus spp., Streptococcus spp. and Corynebacterium spp. ("Amount Of Normoflor"). In patients infected with C. trachomatis, compared with clinically healthy men, the relative number was significantly higher of Bacteroides spp. / Porphyromonas spp. / Prevotella spp., Peptostreptococcus spp. / Parvimonas spp.; in patients infected with N. gonorrhoeae - Anaerococcus spp. and in patients infected with M. genitalium - Megasphaera spp. / Veillonella spp. / Dialister spp., Anaerococcus spp., Peptostreptococcus spp. / Parvimonas spp. and Eubacterium spp. CONCLUSION: An increase in the total bacterial contamination of the urethra in STI was found, most pronounced in infection with Neisseria gonorrhoeae. The best diagnostic indicator that distinguishes normal microbiota from the microbiota of patients with STIs is the sum of Staphylococcus spp., Streptococcus spp. and Corynebacterium spp. In patients with clinical signs of an inflammatory reaction and the presence of STIs, a decrease in the normoflora in all types of STIs and an increase in obligate anaerobic bacteria - Megasphaera spp. / Veillonella spp. / Dialister spp., Bacteroides spp. / Porphyromonas spp. / Prevotella spp., Anaerococcus spp., Peptostreptococcus spp. / Parvimonas spp. and Eubacterium spp.

[The parietal intestinal microflora in patients with chronic pancreatitis].

The article presents the results of the proximal small intestine parietal microbiocenosis research in patients with chronic pancreatitis by polymerase chain reaction in real time. The study includes an assessment of the pharmacological correction's efficiency in this category of patients.

[The composition of the gastrointestinal mucosa-associated microbiota in irritable bowel syndrome patients].

THE AIM OF THIS STUDY: To investigate and analyse the gastrointestinal mucosal-associated microbiota in the samples from diarrhea-predominant IBS (D-IBS), constipation--predominant IBS (C-IBS) patients and healthy controls. METHODS: Oral cavity, duodenal, colonic and rectal mucosal samples were obtained from 40 IBS patients (20 C-IBS, 20 D-IBS) and 14 healthy controls. Duodenal and colonic tissue was collected during a flexible duodenoscopy and colonoscopy. Tissue samples were frozen for further molecular analysis. DNA was extracted from all frozen samples and used to identify 29 specific bacterial groups using quantitative real-time PCR (qPCR). A statistical treatment of the received data was performed. RESULTS: The dominant groups of bacteria of various microbiotops of the gastrointestinal tract in IBS patients and healthy controls were determined. qPCR analysis of duodenal samples demonstrated a reduction in the Bifidobacterium concentration in tissue samples from C-IBS patients when compared to healthy controls (p < 0.05). Analysis of rectal samples demonstrated an increase in concentrations of Faecalibacterium praustnizi (p < 0.05) and a reduction in the concentration of Streptococcus spp. (p < 0.01), Atopobium claster (p < 0.05), Ralstonia spp.+Burkholderia spp (p < 0.05) in tissue samples from D-IBS and C-IBS patients when compared to healthy controls. CONCLUSIONS: Our molecular data indicate that quantitative differences exist in specific bacterial groups in the microbiota between IBS and healthy subjects. The concentration of representatives of Bacteroidetes phylotypes appeared to be the most stable in various microbiotops of the gastrointestinal tract. Small intestinal bacterial overgrowth in IBS patients was not found. Received data help to suggest correlation with the features of the microbiota with clinical form of IBS.

[Immunogenetic aspects of early rheumatoid arthritis].

The study is aimed to investigate the distribution of alleles of HLA-DRB1 gene in patients with early rheumatoid arthritis and healthy individuals in Russian population, and evaluate their significance as molecular genetic markers of rheumatoid arthritis predisposition and protection. The association between alleles of HLA-DRB1 genes, antibodies to cyclic citrullinated peptides and IgM rheumatoid factor was also studied. Low and high resolution HLA-DRB1 genotyping were compared. In the cohort of patients with early rheumatoid arthritis, the alleles of HLA-DRB1 gene were found to be markers of rheumatoid arthritis protection/risk, especially in the homozygous state. They determined production of antibodies to cyclic citrullinated peptides but were not associated with rheumatoid factor IgM levels. These findings support different autoimmune mechanisms of rheumatoid arthritis pathogenesis.

[Immunogenetics of type 1 diabetes mellitus--from fundamental ideas to medical practice].

The review of studies of Russian researchers on theoretical and practical aspects of genetic predisposition to type 1 diabetes associated with immunity: HLA and not HLA genes. Most important for practical public health outcomes are evidence that HLA-genetic predisposition to type 1 diabetes is associated with the DRB1-genotype, consisting entirely of variants DRB1-genes associated with the development of T1D. It was also established that CTLA4 gene has an independent predictive value for T1D.

[The Russian gene pool: gene geography of Alu-insertions (ACE, APOA1, B65, PV92 TPA25)].

The analysis of five Alu insertion loci (ACE, AP4OA1, B65, PV92, TPA25) has been carried out for the first time in 10 Russian populations (1088 individuals), covered all parts of historical area of the Russian ethnos. Depending on locus, Russian populations exhibit similarity with their western (European populations) or with the eastern (populations of the Ural region) neighbors. Considering frequencies of the studied Alu-insertions, Russian gene pool exhibits low variation: average difference between populations is d = 0.007, whereas on classical markers, mtDNA and Y chromosome heterogeneity of Russian gene pool is essentially higher (0.013, 0.033 and 0.142 respectively). Therefore, this set of five Alu insertions has lower variability on the intra-ethnic level. However in inter-ethnic comparisons the clear pattern was obtained: 13 Eastern European ethnic groups formed three clusters, according with their historical and geographical position--East Slavic, Caucasian and South Ural clusters. The obtained data confirms efficiency of using Alu insertions for studying genetic differentiation and history of a gene pool of the Eastern European populations.

[The physiological role of the HLA system and the population aspects].

Identification of HLA genes and further identification of their biological role were among major achievements of medical-biological science in XX century. Several Nobel Prizes awarded in that field of science proved their importance. The end of XX Century and beginning of the XXI century were marked by introduction of molecular genetic methods in research. The contributed to development and revision of current ideas of physiological role of HLA supporting genetic homeostasis and human survival as species. Moreover they prompted to review the existing concepts of inter- and intra-populational polymorphism among HLA-frequencies.

[The use of achievements in human molecular immunogenetics in the management of type 1 diabetes mellitus].

New original data are presented on the use of achievements in human molecular immunogenetics in the management of type 1 diabetes mellitus. They include materials allowing for the prediction of the development of the disease at the population, family, and individual levels along with novel approaches to its radical treatment by the reconstitution of the lost glucose tolerance. The reported data may find wide application in current clinical practice. They open up new prospects for the enhancement of efficacy of prognosis, diagnosis, and treatment of type 1 diabetes mellitus and other autoimmune diseases.

[The distribution of HLA genes in Russian patients with germ cell tumors].

The correlation between DRB1, DQA1, and DQB1 genes of HLA class II, and the development of germ cell tumors (GCTs), as well as serological response to HERV-K proteins were investigated. Genomic DNA prepared from 99 GST patients was subjected to HLA typing by polymerase chain reaction (PCR) using the set of sequence specific primers (PCR-SSP). This set of primers made it possible to detect 14 specificities of DRB 1 locus, 12 alleles and groups of alleles of DQB 1 locus, and 8 alleles of DQA1 locus. Alongside with the definition of the occurrence of HLA markers in the total group of patients, the frequency of the occurrence of HLA-DR-DQ alleles was calculated in: 1) patients with different morphological forms of GSTs (seminomas and non-seminomas); 2) GCT patients producing or non-producing antibodies to Gag and/or Env HERV-K proteins. The comparison group consisted of 300 Moscow blood donors. The study did not reveal statistically significant differences in the frequency of the occurrence of DRB1, DQA1, and DQB1 alleles between the total group of GCT patients, its subgroup, and the control group. Thus, the data obtained demonstrated the absence of a strict correlation between the distribution of HLA class II alleles and GCT occurrence in the Russian population, as well as the ability of GCT patients to develop an antibody to HERV-K proteins, though more numerous observations are required to confirm this conclusion.

[New possibilities of genotypic in the prediction of the likelihood of autoimmune diseases].

The purpose of the study was to estimate the incidence of thyroid dysfunction and cardiovascular diseases in perimenopausal females. The cross-sectional study covered 554 females (mean age 52.6±6.1 years). The levels of thyroid-stimulating hormone (TSH), total cholesterol (TC), high-density lipoproteins (HDL), low-density lipoproteins (LDL), and very low-density lipoproteins (VLDL), the incidence of arterial hypertension (AH), coronary heart disease, chronic heart failure, myocardial infarction, cerebral circulatory disorders, and the severity of menopausal syndrome (MS) were determined. The study detected euthyroldism in 381 (68.8%) patients, hypothyroidism in 168 (30.3%), out of them 35 (20.8%) patients having primary hypothyroidism, and hyperthyroidism in 5 (0.9%) females. Out of the 133 patients receiving L-thyroxine therapy, hypothyroidism was compensated. In 78(58.7%) cases, the dose of L-thyroxine was inadequate. The level of LDL was significantly higher in hypothyroidism; the median of TC was higher than the normal levels in both groups. There were no differences in the incidence of vascular disease between the groups. In both groups, AH was encountered In more than 60% of cases. The females with hypothyroidism had a more severe course of MS. With the adequate dose of L-thyroxine, the level of HDL was significantly higher and that of triglycerides and VLDL was lower than in hypothyroidism. It is expedient to include the measurement of TSH levels into the algorithm of examination of patients with severe MS.

[New ideas of the physiological role of the genes' HLA in reproductive process].

The data presented in this paper shows the role of HLA genes and their products HLA-antigens in reproduction. The study is concentrated on new ideas of the role and mechanisms underlyin the functions of both classical and newly determined HLA genes and their products: New data and hypotheses on HLA-molecules' role in "physiological" pregnancy are discussed.

Acute disseminated encephalomyelitis in children: clinical features and HLA-DR linkage.

This study includes 90 children (41 female and 49 male) in the age range of 2-16 years with acute disseminated encephalomyelitis (ADEM). Thirty-three patients developed ADEM following rubella infection, 26 children following varicella infection, 20 suspected viral aetiology ADEM and 11 multiphasic disseminated encephalomyelitis (MDEM). All patients had neurological, routine laboratory and viral serology study with an enzyme-linked immunosorbent assay. Brain and/or spinal cord magnetic resonance imaging (MRI) were performed in 14 children. A follow-up study was in 1-5 years. Typing of DRB1 gene HLA class II was performed in 38 patients. We established that the varicella and rubella groups had preferential patterns. Rubella ADEM is characterized by acute explosive onset, seizures, coma and moderate pyramidal signs, whereas varicella infection is characterized by cerebella ataxia and mild pyramidal dysfunction. The suspected viral aetiology ADEM was characterized by polisymptomatic presentation. MDEM was characterized by older age of patients (11.6 +/- 2.8 years), more severe and prolonged local neurological symptoms, including myelitis symptoms and marked extrapyramidal signs, with distinct demyelination in MRIs. As a whole, ADEM is associated with DRB1*01 and DRB1*017(03) in the Russian population. Thus, ADEM is a separate autoimmune condition with a specific mechanism due to the type of genetic immunoregulatory base and specificity of viral trigger.

[The risk factors for the development of multiple sclerosis in the Moscow population. II. The combination of exogenous and hereditary factors]. / Faktory riska razvitiia rasseiannogo skleroza v moskovskoi populiatsii. II. Sochetaniia ékzogennykh i nasledstvennykh faktorov.

Simultaneous study of both external and hereditary risk factors of the development of multiple sclerosis (MS) was carried out in Moscow population. As a result of the genotyping of HLA-DR locus on the chromosome 6, there were found markers of the predisposition to MS: by HLA-DR: DR2(15) way, especially, in homozygous heredity and in the absence of such specificity--alleles of DR3 group. The data were confirmed concerning both the elevation of frequency of the alleles of tumor necrosis factor (TNF)a1 and alpha 2 and the decrease of frequency of TNF-alpha 7 allele in MS. There was also confirmed association of the disease with the external factors, such as "predominance of the meat nutrition at the age under 15 years" and "tonsillitis at the age under 15 years". There were two combinations of the external and hereditary factors, when the power of the association increased significantly: "predominance of meat nutrition at the age under 15 years" and DR2(15) and "tonsillitis at the age under 15 years" and DR3. When genetic factors were excluded, the relation of MS with the external factors was considerably reduced; when the external factors were excluded, the association of MS with genetic markers became statistically insignificant. A combination of these external factors, which were the sources for the additional antigenic immunostimulation, with certain genetic factors in DR region suggested the presence of the genetically determined peculiarities of the composition of antigen presentation HLA-DR in the individuals with DR2(15) and DR3. There was no statistically confirmed presence of a combination of the external factors with TNF-alpha alleles which increased additionally MS risk. The presence of immunostimulating antigen-containing external factors may be significant for the influence of TNF-alpha in less degree than for the realization of predisposition in form of the certain composition of HLA molecules.

[Genetic predisposition factors for multiple sclerosis (based on the data from genotyping patients of the Russian ethnic group)]. / Faktory geneticheskoi predraspolozhennosti k rasseiannomu sklerozu (po dannym genotipirovaniia bol'nykh russkoi étnicheskoi gruppy).

Patients' genotyping of Russian ethnic group with multiple sclerosis (MS) was performed for the first time in two loci of the main complex of histocompatibility: in DR HLA class II (gene DRB1) and in the locus of tumor necrosis factors (TNF). There was no difference in the incidence of alleles groups which correlated with some specificity of DR. Meanwhile TNF-a1 and TNF-a9 alleles were encountered significantly more frequently in patients and TNF-a7 in control group. When all the patients and controls examined were divided into groups in dependence on combination of DR and TNF it was found that relations observed between MS and TNF-a7 and TNF-a9 alleles were displayed much more in individuals which carried alleles of gene DRB1, corresponding to DR15 specificity. These are alleles which are known as the main risk factor of MS in Caucasians. The patients with "protective" TNF-a7 allele were characterized by more favorable course of the disease. Thus highly significant genetic markers were revealed for the first time in region of TNF genes which were associated with increased or decreased risk of MS development at least in Russian ethnic group. There was also possibility of their interaction with group of DR15 alleles of DRB1 gene. One of the markers revealed (TNF-a7) occurred to be bound both with decreased risk of MS and with favorable clinical course, which was observed for genetic markers of MS for the first time. Manifestation of one or another property of TNF-a7 marker depends on the presence of alleles of DRB1 gene which corresponds to DR15 specificity.

[A comparative analysis of the proliferative response of blood mononuclears to mitogens]. / Sravnitel'nyi analiz proliferativnogo otveta mononuklearov krovi na mitogeny.

Comparison of two parameters of the peripheral blood mononuclear proliferative response to mitogens (stimulation index, absolute incorporation of radioactive label into proliferating cultures) has shown a discrepancy between the results obtained with the use of these parameters.

[Immunogenetic examination of families with consanguineous marriages]. / Immunogeneticheskoe obsledovanie semei s blizkorodstvennymi brakami.

14 Uzbek cousin-marriage families have been immunologically examined. 13 persons have been identified, who are probably homozygous in HLA-D region (the results of the reaction in mixed lymphocyte culture). The members of cousin-marriage families, as compared to healthy Russian donors, had lower proliferative response to phytohemagglutinin and concanavalin A and decreased indexes of concanavalin A-induced suppression and natural killer activity against K562 targets (in children). At the same time natural killer activity in adults, T- and B-lymphocyte content and IgM, IgG and IgA levels are similar to the respective indexes in Russian donors.