Diagnostic value of microbiome biomarkers of the periodonite microbiome in patients with the association of chronic periodontitis and diabetes mellitus type 2.

The place of high-tech methods of molecular biology in clinical laboratory diagnostics of various diseases and the development of a system of biomarkers as an important component of diagnostic research is currently attracting the closest attention of the scientific community. In this paper, an attempt is made to use high-tech metagenomic analysis to solve problems that arise due to the high frequency of association of periodontal diseases with systemic pathology, in particular, with type 2 diabetes mellitus. The aim of the study was to determine the taxonomic and metabolic features of the microbiome of periodontal tissues in periodontal diseases associated with type 2 diabetes mellitus, as a model of the ratio of local and systemic effects of periodontal pathogenic bacteria. The study included 16S shotgun sequencing of bacterial DNA as part of biological material from periodontal pockets/dentoalveolar furrows of 46 people - 15 patients with chronic periodontitis associated with type 2 diabetes mellitus, 15 patients with chronic periodontitis unrelated to systemic pathology, as well as 16 healthy people in the control group, followed by bioinformatic processing of the data obtained. The obtained data allowed us to establish the taxonomic features of the periodontal microbiome in the association of chronic periodontitis with type 2 diabetes mellitus, which included the predominance of representatives of the families Prevotellaceae and Spirochaetaceae in its composition. The features of metabolic processes in periodontal tissues with the participation of the microbiome were also revealed, which consisted in an increase in the exchange of cysteine and methionine against the background of a decrease in the metabolism of pyrimidine, methane, sphingolipids, and the synthesis of fatty acids, which are of diagnostic value in assessing the condition of patients with type 2 diabetes mellitus.

Cytokine Profile As a Marker of Cell Damage and Immune Dysfunction after Spinal Cord Injury.

This study reviews the findings of recent experiments designed to investigate the cytokine profile after a spinal cord injury. The role played by key cytokines in eliciting the cellular response to trauma was assessed. The results of the specific immunopathogenetic interaction between the nervous and immune systems in the immediate and chronic post-traumatic periods are summarized. It was demonstrated that it is reasonable to use the step-by-step approach to the assessment of the cytokine profile after a spinal cord injury and take into account the combination of the pathogenetic and protective components in implementing the regulatory effects of individual cytokines and their integration into the regenerative processes in the injured spinal cord. This allows one to rationally organize treatment and develop novel drugs.

[Filifactor alocis and its role in the etiology of chronic periodontitis]. / Filifactor alocis i ego rol' v etiologii khronicheskogo parodontita.

The review is devoted to the analysis of modern ideas about the role of bacteria Filifactor alocis in the etiology of chronic periodontitis. The study of these bacteria, discovered in 1985, is complicated by the difficulty of their detection with cultural methods. According to modern researches, the bacteria F.alocis with good reason can be included in the red complex of periodontal pathogens as the most important pathogens of chronic periodontitis. F.alocis is a synergist of such a key pathogen Porphyromonas gingivalis, as well as a frequent satellite of Fusobacterium nucleatum and, somewhat less frequently, Aggregatibacter actinomycetemcomitans. F.alocis is practically not found in healthy people (except for smokers), with a high frequency accompanies the aggressive course of periodontal disease, and also recorded in endodontitis. Due to the ability to participate in the metabolism of arginine, expressed protease activity, a wide range of virulence factors, F.alocis not only colonizes the periodontal tissues, but also significantly affects the formation of the community of periodontal microorganisms (including viruses), contributing to their invasion of epithelial tissues. F. alocis has a number of unique properties, including resistance to oxidative stress conditions in the home defeat, induction of apoptosis of epithelial cells, extracellular matrix degradation of periodontal tissues, activation of proinflammatory cytokines formulation in sites of its presence, suppression of protective reactions of neutrophilic granulocytes, inhibition of the process of complement activation.

"Shielding" of Cytokine Induction by the Periodontal Microbiome in Patients with Periodontitis Associated with Type 2 Diabetes Mellitus.

Periodontal diseases, especially those with polymicrobial etiology, are often associated with type 2 diabetes mellitus, proceeding more severely and affecting the course of diabetes mellitus. Recently, this feature has been associated with the ability of periodontopathogen microflora to cause not only a local infectious process in the oral cavity, but also to interact with the human immune system and induce various systemic effects. We investigated changes in the salivary cytokine profile of patients with chronic periodontitis, associated and not associated with type 2 diabetes mellitus. We observed a statistically significant decrease of MCP-1/CCL2, GM-CSF, IL-5, IL-6, and IFN-γ in the saliva of patients with chronic periodontitis associated with type 2 diabetes mellitus in comparison with patients with chronic periodontitis only. All of these cytokines are associated with macrophage activation. These data are an important contribution to the elucidation of the mechanism of periodontopathogens involvement in the manifestation of the systemic effects of type 2 diabetes.

Metagenomic Analysis of Gingival Sulcus Microbiota and Pathogenesis of Periodontitis Associated with Type 2 Diabetes Mellitus.

Biofilm of the gingival sulcus from 22 patients with type 2 diabetes mellitus and periodontitis, 30 patients with periodontitis not complicated by diabetes mellitus (reference group), and 22 healthy volunteers without signs of gingival disease (control group) was studied by quantitative PCR. Quantitative analysis for the content of P. gingivalis, T. forsythia, A. ctinomycetemcomitans, T. denticola, P. intermedia, F. nucleatum/periodonticum, and P. endodontalis in the dental plaque was performed with a Dentoscreen kit. The presence of other bacterial groups was verified by metagenomic sequencing of the 16S rRNA gene to evaluate some specific features of the etiological factor for periodontitis in type 2 diabetes mellitus. Specimens of the Porphiromonadaceae and Fusobacteriaceae families were characterized by an extremely high incidence in combined pathology. The amount of Sphingobacteriaceae bacteria in the biofilm was shown to decrease significantly during periodontitis. Metagenomic analysis confirmed the pathogenic role of microbiota in combined pathology, as well as the hypothesis on a possible influence of periodontitis on the course and development of type 2 diabetes mellitus.

[MOLECULAR BIOLOGY OF HEPATITIS B VIRUS AND IMMUNOPATHOGENESIS OF CHRONIC VIRAL HEPATITIS B].

Chronic hepatitis B belongs to a category of socially significant diseases due to its wide abundance in the world and high frequency of unfavourable outcomes of this disease. Features of interaction of hepatitis B virus with human immune system, accompanying development of mechanisms of escape from immunological control, is the basis of development of chronic hepatitis B. Molecular-biological features of hepatitis B virus are the basis of the indicated mechanisms, and the content of this review is their examination. Herewith, stages of immunopathogenesis of this disease is the basis of characteristics of interaction of viral proteins with cells of immune system, and isolation of those is accepted in contemporary foreign literature.

[The role of immune factors in the progression of chronic kidney diseases in HIV infection].

AIM: To determine the significance of immune factors in the pathogenesis of kidney injuries in HIV infection, by investigating the cellular and cytokine components of an immune response. MATERIALS AND METHODS: Thirty HIV-infected patients (mean age 31.7±6.2 years) with chronic kidney disease (CKD) were examined. A comparison group consisted of 10 HIV-infected patients without signs of kidney injury. A control group included 24 healthy individuals to analyze immune status and 15 people to estimate the normal values of the cytokine composition. The cellular composition of lymphocytes on a typical immunogram was determined on a flow cytofluorometer; the serum concentrations of cytokines were measured on a multichannel photometer. RESULTS: The HIV-infected patients with kidney injury displayed significant reductions in the absolute (0.2·109/l and 0.4·109/l, respectively; р=0.015) and relative (14.75 and 22%, respectively; р=0.005) counts of CD3+/CD4+ cells and in the immunoregulatory index (0.2 and 0.4, respectively; р=0.014) as compared to those in HIV-infected patients without kidney disease (р≤0.05) with a rise in the number of cytotoxic T cells (CD3+/CD8+). The HIV-infected patients showed a preponderance of immunosuppressive cytokine compositions, as indicated by the high levels of transforming growth factor-ß (a more than 50-fold increase) and by a statistically significant rise in the level of tumor necrosis factor-α (TNF-α) (with CD4+ lymphocyte counts more or less than 200 cells/µl - 19.0 and 24.2 pg/ml, respectively; p=0.017; with HIV RNA levels more and less than 100,000 copies/ml - 24.4 and 19.7 pg/ml, respectively; p=0.012). CONCLUSION: The HIV-infected patients with CKD developed kidney injury in the presence of a more pronounced decrease in blood T helper lymphocyte subpopulation levels with a predominance of proinflammatory and immunosuppressive responses. TNF-α in combination with immunosuppression and high viral loads was established to play a leading role in the development of kidney injury in HIV infection.

[MECHANISMS OF INTERACTION OF VIRAL CAUSATIVE AGENTS IN PATIENTS CO-INFECTED WITH HUMAN IMMUNODEFICIENCY AND HEPATITIS C VIRUSES].

In patients infected with human immunodeficiency virus (HIV) in 20 - 30% of cases co-in- fection with hepatitis C virus (HCV) is observed, that is associated with common routes of transmis- sion for these causative agents. The main cause of lethal outcome for co-infected patients is liver damage. Thus, analysis of mechanisms of mutual influence of HIV and HCV under the conditions of co-infection gains special attention, that can be examined from both standpoints of direct inter- molecular interaction of 2 viral causative agents, as well as from the position of their immune- mediated effect. Negative effect of HIV on the course of fibrosis process in liver during HCV infection is associated with the feature of this virus to cause deep alteration in the immune system by direct damage of CD4+ cells, disruption of mechanisms of immunological memory, suppression of functions of liver fraction of NK and NKT, as well as its ability of co-receptor interaction with hepatocytes and stellate cells, enhancing progress of fibrosis alterations and HCV replication in liver. HCV.is also established to effect replication of HIV, stimulate infection of macrophages with this virus. All these events facilitate the rise in lethality during HIV and HCV co-infection.

[Bronchial asthma pathogenesis and genetic prognosis development].

The review is dedicated to an actual problem--genetic prognosis of risk of bronchial asthma development that is quite a complex aspect of studies from a methodological viewpoint. Bronchial asthma--heterogeneous disease by both etiology and clinical characteristics. At the same time genetic prognosis is based on the unity of pathogenetic mechanisms of development, though in immunological reactions that are the base of this disease, alternative variants are possible. The aim of this review is carrying out parallels between modern achievements in the field of deciphering trigger mechanisms of bronchial asthma pathogenesis and object of genetic studies based on these mechanisms. Among the examined conceptions--role of epithelial tissue in trigger mechanisms of bronchial asthma, variants of key role of immune system cells, first of all, T-helpers of various types for further development of inflammatory-effector reactions with damage characteristic for this disease. Compliance of contemporary approaches of genetic studies and novel concepts of bronchial asthma pathogenesis is shown.

[Oral infections].

In the survey the basic aspects of the infectious pathology are analyzed, that are to be met in the work of stomatologist. There is in detail described the intra-hospital infection, caused by conditionally pathogenic flora. There is given the clinical characteristics of the oral manifestations of the basic of <> infectious diseases.

[Features of proliferative response of T-cells on hepatitis c virus antigens in healthy individuals contacting with this virus].

AIM: Evaluation of CD3+ and CD3-/CD56+ proliferative response on hepatitis C virus antigens in healthy medical workers. MATERIALS AND METHODS: The study included 15 medical workers with length of service of 2 and more years without common risk factors (blood and blood products transfusion, abdominal operations, invasive procedures, use of intravenous drugs). Control group consisted of 9 healthy individuals without risk factors. Peripheral mononuclears were isolated from blood and then incubated 72 hours in the presence of mitogen/PHA, Core+NS4 1b genotype HCV, NS4 HCV2a+3a genotypes or medium. Proliferative activity was registered by the presence of cell division marker ki67 by using FACS. RESULTS: The initial immunogram of medical workers differed from the control group by a significantly lower quantity of CD3+ lymphocytes, in particular CD3+/CD8+ population. Incubation with PHAresulted in a decrease of quantity of CD3+/ ki67+, CD4+/ki67+ and CD3-/CD56+/ki67+ in the medical workers group. Cultivation with HCV antigens resulted in a significant decrease of Treg (CD3+/CD25high/FoxP3+) and activated T-lymphocytes population in the case of stimulation by Core and NS4 1b genotype antigens. Analysis of cell response on virus antigens based on proliferative activity index detected significant differences only for CD8+/ki67+. Stimulation by Core and NS4 1b genotype antigens resulted in an increase of quantity of these cells whereas in the case of NS4 2a+3a genotypes their decrease was observed. CONCLUSION: The described changes may reflect exhaustion of cell reactivity due to extra antigen load in the group of medical workers, while differentiated immunologic shifts on hepatitis C viruses of various genotypes are noted.

[Intestine infections, inflammation and autoimmunity. Lymphoid apparatus of intestine in interaction with intestine microflora].

Topicality of interrelation between intestine infections, inflammation diseases of intestine and autoimmune processes is widely discussed in scientific literature of recent years. Thereby a review of literature on the designated aspect of the problem is dedicated to the analysis of interconnection between structural-functional features of lymphoid apparatus of intestine and its ability to react to antigen load from both commensal and pathogenic intestine microflora. During description of structure and functions of lymphoid formation of intestine a particular attention is paid to difference of subpopulation characteristics of lymphocytes and antigen-presenting cells composing intra-epithelial lymphocytes, elements ofimmune system lamina propria, Peyer's patches, mesenteric lymphatic nodes. The role of normal microflora and infectious agents in trigger mechanisms of reaction of immunocompetent cells is underscored; key aspects of cellular-molecular mechanisms of mucous membrane immune system functions are discussed.

[Intestine infections, inflammation and autoimmunity. Trigger and effector mechanisms of autoimmune disease development as an outcome of intestinal infections].

Problem of interconnection of intestine infections, inflammatory intestine diseases and autoimmune illnesses in this article is examined from the position of their trigger and effector mechanisms. Among trigger mechanisms special attention is given to mechanisms by which the presence of pathogenic microbial causative agent in the organism is transformed into an autoimmune process. The phenomenon of antigen mimicry, carriage of superantigens by pathogenic agents, the role of cell apoptosis are accentuated. Autoimmune diseases are examined in the same way as genetically determined phenomenon with designation of main genes, polymorphism of which is involved in the development of this pathology. Among effector reactions accompanying the development of autoimmune process againstthe background of intestine infections the role of B1 lymphocytes, Th17 and Th1 are analyzed in more detail. Special attention is given to pathogenetic and protective role of natural killers which is recognized as relatively understudied.

[Serotonin and its immune and physiological effects].

Now that the neurotransmitter serotonin modulates the immune system cells, and its main sources for antigenpresenting cells and lymphocytes are enterochromaffin cells of the gut, peripheral nerves, platelets and mast cells in case of inflammation. Immune cells uptake serotonin because they express receptors for this monoamine and intracellular serotonin transporters. The dendritic cells have a mechanism to transfer serotonin to T lymphocytes during antigen presentation. The macrophages and T cells have the ability to serotonin synthesis. Serotonin can influence mobility and proliferation of lymphocytes, phagocytosis, cytolytic properties, synthesis of chemokines and cytokines. Diversity of immunomodulating effects of serotonin is determined by heterogeneity of serotoninergic receptors. Immunomodulating action of serotonin is evidence of the close relationship between nervous and immune systems.

[Immunogenetic predictors of a rapid virologic response to antiviral therapy in patients with chronic hepatitis C].

AIM: To determine the correlation between interleukin 28B (IL28B) gene polymorphism in patients with chronic hepatitis C (CHC), the presence or absence a rapid virologic response to antiviral therapy, and a number of immunological characteristics as a basis for a personalized approach to treating the patients. SUBJECTS AND METHODS: Seventeen CHC patients infected with hepatitis C virus genotype 1b were examined and underwent genetic testing for IL28B gene polymorphism for rs12979860 (CC, CT or TT genotypes) and rs8099917 (TT, TG or GG genotypes) using the modified method of adjacent samples, which revealed single nucleotide substitutions in the genes. Their immunological parameters were identified by a flow cytometry technique by taking into account whether a rapid virologic response had been achieved. RESULTS: The key phenomena of a rapid virologic response in the representatives of different IL28B genotypes are the nonspecific proliferative activity of blood natural killer cells before treatment, as well as the count of regulatory T cells before and 4 weeks after therapy start. CONCLUSION: To predict the efficiency of antiviral therapy for CHC, it is desirable to supplement genetic studies with immunological data.

[Proteomic technologies for identification of serum potential biomarkers of autoimmune demyelinating polyneuropathies].

Time-of-flight MALDI mass spectrometry (MALDI-TOF-MS) profiling of blood serum of patients with Guillain-Barré syndrome (GBS, 36 samples), chronic inflammatory demyelinating polyneuropathy (CIDP, 24 samples) and practically healthy donors (HD) (35 samples) was carried out in order to identify potential biomarkers of autoimmune demyelinating polyneuropathies (ADP). To simplify the peptide-protein mixture of serum prior to MALDI-TOF-MS analysis samples were pre-fractionated on magnetic microparticles with a weak cation-exchange (MB-WCX) surface. Comparative analysis of mass spectrometric data using the classification algorithms (genetic and neural network-controlled) revealed a characteristic set of peaks, agreed change area with a high specificity and sensitivity of the differentiated mass spectrometry profiles of the blood serum of patients with DPNP and healthy donors (for GBS values of these characteristics reached 100 and 100, and for CIDP 94.1 and 100% respectively). Comparative analysis of mass spectrometric profiles of serum samples obtained from patients with GBS and CIDP, allowed to build a classification model to differentiate these diseases from each other, with a specificity of 88.9 and a sensitivity of 80%.

[Immunological aspects of HCV/HIV coinfection].

AIM: To evaluate effects of HCV/HIV coinfection on blood lymphocyte phenotype including cells CD56+. MATERIAL AND METHODS: We studied standard immunogram, subpopulational composition and functional activity of blood cells CD56+ in 67 patients with verified diagnosis of HIV infection and virus hepatitis C. RESULTS: We discovered some pathogenetically sound indices of immune status in patients with HCV/HIV coinfection. Some of the immune shifts detected changed in response to intake of narcotic drugs, antiretroviral treatment, hepatic cirrhosis, pneumonia. CONCLUSION: Parameters of subpopulational composition and functional activity of cells CD56+ in the blood of patients with HCV/HIV coinfection elucidate some unknown features of the above infectious process which should be considered in this coinfection treatment and prophylaxis.

[Association between expression of lectin type receptors by natural killers and intensity of liver fibrosis during chronic hepatitis C].

AIM: To study functional activity of natural killers on different stages of fibrosis during chronic hepatitis C. MATERIALS AND METHODS: Functional activity of CD3-/CD56+/CD16+ lymphocytes measured as expression of natural killers receptors (NKR) and natural cytotoxicity receptors (NCR) was assessed by flow cytometry. RESULTS: At stage I of fibrosis, decrease of number of CD3-/CD56+/NKG2D+ cells was observed, whereas at precirrhotic stage III--sharp decrease of CD3-/CD56+/CD94+ and CD3-/ CD56+/NKG2D+ populations, and at cirrhotic stage--decrease of number of CD3-/CD56+/ NKG2D+ cells and increase of cytolytic activity of natural killers carrying CD107a marker compared to precirrhotic stage. CONCLUSION: Obtained data demonstrate that natural killers during chronic hepatitis C receive regulatory signals mainly through lectin type receptors (CD94 and NKG2D).

[Clinical and prognostic criteria for different forms and types of the course of Yersinia infection].

AIM: to reveal the prognostically significant symptom complexes of pseudotuberculosis/yersiniasis in their different forms and types. SUBJECTS AND METHODS: A comprehensive examination and a long-term follow-up were made in 295 patients with yersiniasis and pseudotuberculosis. A special score scale was developed to evaluate the prognostic value of their clinical symptoms. RESULTS: The symptom complexes prognostically significant to the outcome of Yersinia infection were found. CONCLUSION: The use of the described prognostically significant symptom complexes of Yersinia infection makes it possible to correctly evaluate the course of the disease in each case, identify risk-group patients, and to use adequate treatment.