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Rheumatoid arthritis (RA) is a chronic, systemic, inflammatory, autoimmune disease, the development of which is associated with impaired mechanisms of cell number control, proliferation balance, and apoptosis. The aim of our study was to perform a comprehensive analysis of cellular and humoral effects exerted by “apoptotic” T lymphocyte culture upon “proliferating” cells, taking into account the parameters of “primary” and “secondary” apoptosis induction, activation and proliferation markers, viable cell contents, role of cortisol, caspase and cytokines during cultivation under the “cell neighborhood” conditions in healthy subjects and patients with RA. The results of the analysis, with respect to previous data, revealed both similar and different functions of the T cell systems generated under “cell-neighborhood” conditions in healthy individuals and RA patients. Both groups under study were characterized by secondary induction of apoptosis, which refers to non-autonomous effects, along with appearance of activated T lymphocytes at significant amounts, increase of IL-6 and IL-4 levels, as well as low levels of IFNγ and caspase 8, decrease potential of receptor and mitochondrial apoptosis. No differences in apoptosis level were found between donor and patient groups. At the same time, the initially high level of TNFα in healthy subjects decreased during the induction of apoptosis, along with moderate increase of IL-6. The levels of intracellular p53 and Bcl-2 molecules did not change, and many positive correlations remained between the studied factors in all variants of mixed and “apoptotic” cultures, thus suggesting maintenance of functional T cell balance under the unfavorable conditions in donors. The RA patients were characterized by increased number of live cells in “apoptotic” cultures, a significant increase in the number of Ki-67+ T lymphocytes, being indicative for proliferative processes under conditions of apoptosis, like as absence of TNFα response to apoptosis induction. Lack of relations between the studied molecules in “apoptotic” cultures and local cortisol synthesis, as well as presence of correlations between cortisol and the studied molecules in apoptotic cultures, presumed some changes of intercellular interactions and disturbance of homeostasis among T lymphocytes under the conditions of “cellular neighborhood” were also observed in RA patients.

Psoriatic arthritis (PsA) is a chronic immune-mediated inflammatory joint disease, often associated with psoriasis. Interactions between immune cells, mainly T lymphocytes, and cells of the osteoarticular system are central in the pathogenesis of PsA. Th1, Th17 cytokines associated with the development of PsA contribute to an increase in the production of cytokines, chemokines, matrix metalloproteinases, adhesion molecules by immune cells, chondrocytes, fibroblasts, and induction of osteoclasts. That leads to the destruction of cartilage and bone tissue. Among the cytokines potentially involved in the pathogenesis of PsA, IL-7 is of particular interest. IL-7 is a T cell survival factor. However, it can promote the production of IFNγ and TNFα by T cells. IL-7 may indirectly promote osteoclast maturation and cartilage degradation. The aim was to investigate the effect of IL-7 and blockade of the α-chain of the IL-7 receptor (IL-7R) in vitro on the production of IL-5, IL-13, IL-2, IL-6, IL-9 IL-10, IFNγ, TNFα, IL-17A, IL-17F, IL-4, and IL-22 by T cells in norm and PsA.The study included 14 patients with PsA in the acute stage of the disease and 8 healthy individuals. To determine cytokines concentration, multiplex analysis was performed using flow cytometry.It was shown that IL-7 enhances the production of Th1, Th2, Th17, Th22, and Th9 cytokines in both patients with PsA and healthy individuals. The exception was IL-2 for both groups. Under the blockade of IL-7R with monoclonal antibodies, the level of IL-6 increases and production of IFNγ, TNFα, IL-17F, IL-10, IL-5, and IL-9 by donors’ cells and production of IFNγ, TNFα, IL-22, IL-2, IL-4, IL-5, IL-13, and IL-9 by cells from patients with PsA decreases relative to production of cells stimulated with IL-7. In donors, the blockade contributed to a change in the balance of Th1/Th2 cytokines: level of IL-4, IL-13 did not change on the background of a decrease in production of IFNγ, TNFα. In patients with PsA, under blockade of IL-7R the production of IL-10 remained at an increased level and concentration of IFNγ, TNFα and IL-2, which are actively involved in the tissue damage mechanisms, decreased. The obtained data indicate the prospect of using the IL-7R as a target for the treatment of psoriatic diseases.

The aim of the work was to study the species diversity of pathogenic and non-pathogenic hantaviruses circulating in populations of small mammals in the Republic of Bashkortostan using molecular-genetic methods.Materials and methods. Individual samples from small mammals were tested by the nested PCR using genus-specific primers that amplify the L segment of hantaviruses. The resulting PCR products were sequenced by the Sanger’s method from internal nested PCR primers. For samples containing Puumala virus, fragments of the S, M, and L segments of the viral genome were sequenced using Sanger’s method. The construction of phylogenetic trees was carried out using the MEGA X software.Results and discussion. Out of 300 examined samples of small mammals collected on the territory of the Republic of Bashkortostan in 2023, 14 samples have been found positive for the presence of hantavirus RNA: Seewis (8), Tula (3), Puumala (3). The circulation of the non-pathogenic hantavirus Seewis and the opportunistic hantavirus Tula has been established for the first time in the Republic of Bashkortostan. The circulation of the Seewis hantavirus has been confirmed in populations of the common shrew (Sorex araneus) and the pygmy shrew (S. minutus); the Tula hantavirus – in populations of the common vole (Microtus arvalis). Results of phylogenetic analysis substantiate the reassortment origin of one of the genetic variants of the Puumala hantavirus on the territory of the Republic of Bashkortostan. The prerequisites for the formation of combined natural foci of hantaviruses Puumala, Seewis, and Tula on the territory of the Republic of Bashkortostan are discussed.

Arthropathy is one of the most prevalent diseases, which are based on the destruction and remodeling of cartilage and bone tissue. The inflammation that precedes destruction can be caused by mechanical stress on the joints, or by autoimmune reactions. Recently, IL-7 is considered as one of the key cytokines that promote the production of matrix metalloproteinases, catabolic enzymes, T cell-mediated activation of monocytes, and maturation of osteoclasts. The soluble form of the IL-7 receptor can help prolong the lifespan of IL-7 and thereby it ensures the bioavailability of the cytokine and mediates effect of IL-7 on cells. The aim of this study was to determine the soluble form of the IL-7 receptor (sIL-7R) in the blood plasma of patients with rheumatoid arthritis (RA), osteoarthritis (OA), psoriatic arthritis (PsA) and psoriasis vulgaris (PS), as well as healthy individuals. The RA patients included in the study had moderate to high disease activity according to the DAS28 index. Patients with PsA predominantly had moderate and low disease activity (DAS28) and were characterized by mild to moderate disease severity (PASI). In accordance with the PASI index, patients with PS with mild and severe severity of the disease were included in the study. All patients with OA had a metabolic phenotype that is accompanied by an elevated body mass index.sIL-7R was determined in blood plasma by enzyme-linked immunosorbent assay. It was found that in patients with arthropathy, the level of soluble form of IL-7 was increased relative to healthy individuals, with the exception of the group of patients with PsA. Also, a high concentration of sIL-7R was observed in patients with PS. Analyzing the clinical characteristics of the patients, we found that sIL-7R levels were elevated in RA and PsA patients with high disease activity by DAS28. In addition, positive correlations were found between the concentration of sIL-7R and DAS28 in RA and PsA. In patients with PsA with moderate severity of the disease (PASI), the concentration of sIL-7R was also increased relative to donor's values. On the contrary, in patients with PS, a high level of sIL-7R was noted regardless of the severity of the disease. In patients with OA, no relationship was found between sIL-7R levels and clinical parameters.Thus, an elevated level of sIL-7R in patients with arthropathy may indicate the involvement of IL-7 and its receptor system in the pathogenesis of joint diseases. The IL-7 receptor may become a promising target both in the treatment of joint diseases and other autoimmune diseases, including psoriasis.

The gut is inhabited by a trillion bacteria that produce up to 60% of the host’s metabolites. The gut microbiome plays an important role in regulating host immune function. A lot of research concerned the effect of probiotic on the pathologies associated not only with dysbiosis and metabolic disorders, but there is breakthrough in the treatment of inflammation, oncology and neurodegenerative disorders. Animals with mutation of the genes leading to pathology used to assay probiotic effect. To understand direct action of probiotics, cells derived from control mice or cell culture of tumor genesis in vitro studies are used. However, there is little research of the probiotic effect on cells derived from mice with pathology. In this study, we assessed the phenotypes of dendric cells derived from Muc2-/- mice with chronic inflammation and assessed the effect of L. johnsonii on the dendric cells. It is known that the key features of IBD models are thinning of mucin layer and changes in the intestinal microbiome. We compared the efficiency of maturation and activation of dendric cells derived from the bone marrow of Muc2-/- mice and dendric cells obtained from healthy C57BL/6 mice free from specific species pathogens. We evaluated the expression of co-stimulatory molecules, the proliferative index, and the ability to trigger the T regulatory response of dendric cells, which were stimulated with the probiotic L. johnsonii. Markers of dendritic and T cells were assessed by flow cytometry using antibodies to extra- and intracellular proteins. The proliferative activity of splenocytes was assessed using the WST test. It was shown that dendric cells derived from the Muc2-/- had an immature phenotype. Dendric cells of Muc2-/- mice could not effectively stimulate the proliferation of allogeneic and syngeneic T cells. L. johnsonii was able not only to stimulate the maturation of dendric cells derived from Muc2-/- mice, but also to increase the expression of FoxP3 on CD25+ T cell that were co-cultured with DCs. Thus, we believe that this probiotic bacterium can reduce signs of inflammation and reduce pathological processes in animals of an experimental model of IBD in vivo.

A formal description of the mathematical model of a steganographic system based on key information in the form of stegosets is given. The mathematical model of the steganographic system is intended both for placing hidden marks to control the integrity of documents acting as a container, and for covert data transmission. Exist-ing mathematical models do not fully meet the requirements for steganographic systems. The proposed model is based on the concepts of a three-level key, a stegoset, a container component, and a message block. The three-level key is a set of actual steganographic transformations depending on the type of container. A stegoset is an element of a key set. A component of a container is such a subset of it that there is a stegoset for it. A message block is such a sequential part of the message that there is a container component for it, in which it can be hidden using the appropriate stegoset. The model implements the application of different stegosets to different container compo-nents, and the message can be divided into blocks. The model allows controlling the integrity of the message and adapts to different types of containers. Possible options for the practical application of the mathematical model are proposed, such as monitoring the integrity of the message when calculating the control number, splitting the message into blocks and sequentially monitoring their integrity by analogy with the blockchain system.

Cellular homeostasis in the body is known to be maintained by the processes of cell proliferation and death, whereas apoptosis is the most frequent and physiological, “silent” mechanism of cell elimination. It has been currently shown that the process of apoptosis traditionally considered an autonomous event, has a pronounced non-autonomous effect on migration, proliferation, and death of the neighboring cells. This work was based on the data on impaired programmed death of mononuclear cells from the patients with rheumatoid arthritis (RA) leading to the evolving autoimmune inflammation. The aim of this study was to evaluate effector molecules exerting autonomous and non-autonomous influence of T cell apoptosis under the conditions of “cell neighborhood” in cell cultures of healthy people and RA patients. The studies were performed with blood samples of RA patients and healthy women of comparable age. These experiments were performed in order to assess the levels of main molecules mediating the in vitro receptor and mitochondrial apoptosis of T lymphocytes. In previous studies, using the original “cell neighborhood” model, no differences were found in parameters of early and late activation apoptosis between the groups of donors and RA patients. At the same time, 1-week incubation in apoptotic cultures of the patients was followed by significantly increased number of viable cells carrying the proliferation marker Ki-67. Different results of in vitro apoptosis induction in cultures under similar conditions of “cell neighborhood” in healthy people and patients with RA have revealed the importance of main effector molecules of apoptosis in the studied groups. In this study, we have revealed low potential of the receptor pathway for apoptosis activation in healthy people, due to suppression of TNFα production during cell incubation under the conditions of “cell neighborhood”, and in RA patients due to initially low TNFα in supernatants which did not change over time and in various incubation variants, along with low content of initiating caspase 8 in both groups. Significant suppression of effector molecules of mitochondrial pathway of apoptosis activation, i.e., Bcl-2 anti-apoptotic factor and p53 transcription factor was detected in cultures of apoptotic cells, as well as mixtures of proliferating and apoptotic cells under the conditions of “cell neighborhood” in RA patients. The amounts of these molecules did not change in healthy persons. At the same time, no differences in these molecules were found between individual variants of cell cultures from the patients with RA and healthy people. The both studied groups were characterized by a significant activation of IL-4 and IL-6 production, i.e., the cytokines with autonomous and non-autonomous protective and reparative properties, Hence, one may conclude that high levels of these cytokines had different effects in cell cultures under the conditions of “cell neighborhood”. Incubation of cells from healthy people under suboptimal conditions was associated with maintaining the balance of proliferation and apoptosis, whereas, in cell cultures of RA patients, this balance caused activation of proliferation processes, being accompanied by an increase in the number of living cells in apoptotic cultures.

Aim. To assess the impact of autologous activated T-lymphocyte immunotherapy on clinical parameters and quality of life in patients with allergic bronchial asthma (BA) in comparison with patients with allergic BA who received standard therapy.Materials and methods. A non-randomized, pilot study included 19 patients with allergic BA of moderate severity (7 men and 12 women aged 23–61 years, average age – 38.5 ± 4.3 years) who received the T-cell vaccine (n = 12) and standard therapy with inhaled glucocorticoids, short- and long-acting β2-adrenergic agonists (n = 7). After signing an informed consent, the patients were subcutaneously injected with autologous activated T-lymphocytes with a frequency of 4 injections 1 time / week, and then 6 injections 1 time / month. The research methods included asthma control measurement according to the ACQ-5 questionnaire and quality of life assessment according to the AQLQ(S) questionnaire. Clinical data were collected during lung function tests and by measuring the total immunoglobulin E (IgE) level.Results. In the course of the study, the immunotherapy was well tolerated, no systemic adverse reactions were noted. The treatment approach in the patients who received the T-cell vaccine resulted in significant improvement of asthma control parameters (according to the ACQ-5 questionnaire) and parameters of the patients’ quality of life (according to the AQLQ(S) questionnaire) within all 4 categories. Besides, their lung function improved by the end of treatment, and the total IgE level decreased. No significant changes in these parameters were observed during the follow-up in patients who received standard therapy. The study was conducted before immunotherapy, after 2 months (after 5 injections), and after 7 months (after 10 injections).Conclusion. Evaluation of the impact of immunotherapy with autologous activated T-lymphocytes on the clinical parameters and quality of life in patients with BA indicates effectiveness of treatment in patients with allergic BA.

Transcriptional activity of genes involved in maintaining genetic homeostasis (genes for repair, cell cycle and apoptosis: TP53, MDM2, ATM, BAX, BCL-2, CDKN1A, OGG1, XPC, PADI4, MAPK8, NF-KB1, STAT3, GATA3) was studied in chronically exposed persons with an increased intensity of early and late stages of apoptosis and necrosis of peripheral blood lymphocytes. The object of this study was peripheral blood mononuclear cells obtained from 132 chronically exposed residents of the Techa riverside villages. The mean accumulated dose to red bone marrow was 426.4±48.2 mGy (1.3–2930.0 mGy), to thymus and peripheral immune organs, 58.9±7.9 mGy (0.1–489.0 mGy). The study was performed more than 60 years after the onset of exposure, the average age of exposed persons was 68±0.6 years (55–86 years). The study of apoptotic and necrotic death of peripheral blood lymphocytes was based on the presence of phosphatidylserine on the cell membrane surface, as well as on its permeability for DNA-intercalating dye. Evaluation of the relative content of mRNA genes for repair, cell cycle, and apoptosis was carried out using real-time PCR. An increased relative content of PADI4 gene mRNA was registered in the group of chronically exposed persons with the increased intensity of early apoptosis (p = 0.006). Modulation of the relative content of mRNA of the TP53 (p = 0.013) and BCL-2 (p = 0.021) genes was detected in the group of chronically exposed individuals with the increased intensity of the late stage of apoptosis. A statistically significant increase in the transcriptional activity of the TP53 gene was observed in the group of chronically exposed persons with the increased intensity of peripheral blood lymphocyte necrosis in the long-term period (p = 0.015). In the course of the study it was noted that exposed people with increased intensity of apoptosis, first of all, demonstrate changes in the transcriptional activity of apoptotic genes. These data are consistent with current views on the activation of programmed cell death.

Psoriasis is a chronic autoimmune disease in which the skin and joints are involved in the pathological process. It was found that the recurrence of rashes in this disease occurs due to the resident memory cells of the skin. The number of CD4+CCR3+ effector memory cells in peripheral blood correlates with the severity of the disease. Therefore, the aim of our work is to study the phenotype of peripheral blood memory cells in patients with psoriasis.The study included 6 healthy donors: average age – 45.4 (min – 29, max – 55), women – 3, men – 3; 10 patients with psoriasis: women – 4, men – 6, average age – 37.3 (min – 23, max – 57), of which 5 patients with PASI > 10 and 5 patients with PASI < 10. The exclusion criteria for the study were the presence of autoimmune, oncological and hematological diseases, systemic therapy with immunosuppressive drugs for 1 month. Patients signed informed consent to participate in the study. Isolation of peripheral blood mononuclear cells was performed in a density gradient of ficoll-urographin (p = 1.082 g/L). Then cells were stained with fluorochrome-conjugated monoclonal antibodies to surface markers of central (Tcm) and effector (Tem) CD4+ memory cells (CD4, CD45RO, CD197), the α-chain of the IL-7 receptor (CD127), and the γ-chain of the IL-7 receptor (CD132). Statistical analysis of the data obtained was performed using the Statistica 6.0 software package.The percent of Tcm in the peripheral blood of donors was 33.4% (in – 18.2, max – 43.7), Tem – 28.7% (min – 13.6, max – 38.9), in patients with psoriasis: Tcm – 28.65% (min – 13.3, max – 59.6), Tem – 21.5% (min – 9.3, max – 38.6). In the peripheral blood of patients with psoriasis, among the central CD4+ memory cells, the proportion of CD127+CD132- -cells is 26.00%, CD127+CD132+ – 1.69%, CD127+CD132- – 69.00%, CD127- CD132+ – 1.94%. Among effector CD4+ memory cells, the proportion of CD127+CD132- -cells is 23.58%, CD127+CD132+ – 1.18%, CD127+CD132- – 69.84%, CD127- CD132+ – 0.70%. A direct correlation was found between the number of CD127- CD132+ central memory cells and the PASI value (r = 0.639, p < 0.05).In patients with psoriasis, the proportion of central memory cells is higher than in healthy donors, while the number of effector memory cells is lower. A direct correlation was found between the number of central cells expressing the γ-chain of the IL-7 receptor and the severity of the disease. Activated memory cells are characterized by high expression of CD132. It can be assumed that this population of memory cells plays a role in maintaining autoimmune inflammation in patients with this disease, and also participates in the repopulation of skin resident memory cells.

Much attention of researchers is directed to study the role of factors of homeostatic proliferation in the pathogenesis of autoimmune diseases, however, the effect of IL-7 and IL-15 on the phenotype of naive T cells has not yet been sufficiently investigated in the pathology. The aim of this study was to investigate the proportion of CD45RA+ and CD31+ naive cells among CD4+ and CD8+ lymphocytes from healthy individuals and patients with rheumatoid arthritis (RA) upon stimulation with IL-7, IL-15 in vitro. In peripheral blood, we did not find any differences in the number of CD45RA+ and CD31+ cells between the group of donors and group of RA patients. In donors, stimulation by a combination of IL-7 with IL-15 promoted an increase in the proportion of CD4+CD45RA+ and CD8+CD45RA+ relative to their level in the peripheral blood. Whereas in RA patients the number of CD8+CD45RA+ cells decreased under IL-15 and the combination of IL-15 with IL-7 compared to the control without stimulation, and it, as a proportion of CD4+CD45RA+ cells, significantly differed from the content of these cells under the same conditions in donors. There were no significant differences in the content of CD31+ cells and the number of CD31+ cells proliferating to cytokines, both between the groups of donors and patients with RA, and between different culture conditions. Thus, we can say that under the factors of homeostatic proliferation, there is a proportional increase in CD31+ T cells number, both in donors and in patients with RA. At the same time, naive T cells from donors retain the expression of CD45RA during cultivation, while naive T cells from patients partially lose it. The obtained data indicate that in RA, under factors of homeostatic proliferation the phenotype of naive T cells is converted into the phenotype of memory T cells.

The process of apoptosis is known that play an important role in cellular homeostasis, and the altered cell death may lead to development of pathological disorders. Evolving autoimmune conditions, in particular, rheumatoid arthritis, are associated with decreased rates of apoptosis as a form of programmed cell death. The aim of this study was to evaluate expression of activation and proliferation markers on T lymphocytes during initiation of apoptotic cell death under the conditions of “cell neighborhood” in healthy individuals and patients with rheumatoid arthritis. Patients and methods. The study was performed with blood samples of the patients with rheumatoid arthritis (RA) and healthy women of comparable age. During the study, we conducted experiments aimed to identify the in vitro influence of non-stimulated apoptosis-induced cells, as well as aCD3- and dexamethasone (Dexa)-stimulated apoptosis-induced cells upon autologous T lymphocytes cultured under physiological conditions. Development of a “cell neighborhood” model, i.e., co-cultures of CFSE- T cells subjected to incubation under crowding condition and depletion of the culture medium which is the most physiological variant of apoptosis activation, and CFSE+ autologous cells placed in the complete culture medium, has revealed some relationships. We have revealed an opportunity of secondary induction of early and late apoptosis by means of humoral and cellular components of autologous cell culture subjected to activation apoptosis. We determined the features of apoptosis in unstimulated, as well as aCD3- and dexamethasone-stimulated cultures, compared with controls. There were no differences in these parameters of apoptosis between RA patients and healthy people for all variants of cultures. An increased proportion of viale cells was found in the CFSE- culture of patients with RA when compared to donors. The donor group had more lymphocytes with activation parameters CD25+, CD69+ and low level of proliferation marker Ki-67 than patients. In contrast to healthy, the RA patients demonstrated a significantly increased expression of Ki 67 in T lymphocytes when co-culturing CFSE- and CFSE+ cells. An increased number of living cells in apoptotic cultures of patients with RA relative to healthy people, in absence of significant differences in the parameters of apoptosis and activation markers in dynamics, as well as pattern of changes in the Ki-67+ cell contents suggested a contribution of the non-autonomous effects of apoptosis to cellular homeostasis in RA patients.

IL-7 is a key factor for the survival and maintenance of CD4+ central (Tcm) and effector (Tem) memory cells in the whole body. In many autoimmune diseases, an elevated level of IL-7 is detected in blood serum and at the site of inflammation, thus suggesting participation of this homeostatic factor in the survival of memory T cells, including auto-reactive clones, in inflammatory disorders. The aim of the study was to investigate the mechanisms of maintaining CD4+ memory T cells under normal and inflammatory conditions. We developed an in vitro model of inflammation, based on induction of pro-inflammatory cytokines, and then evaluated the effects of IL-7 upon purified sorted populations of CD4+Tcm and Tem under normal conditions and in vitro inflammatory model. IL-7 treatment promoted maintenance of CD4+Tcm phenotype in all variants of cultures. In the absence of contact with adherent cell fraction, the IL-7-induced proliferation of Tcm and Tem was slightly reduced, both under normal and inflammatory conditions, thus suggesting low sensitivity of memory T cells to contacts with MHC, and, probably, a requirement for additional signals to provide complete stimulation with IL-7. The last suggestion is also supported by data about CD127 and CD132 expression, i.e., in the absence of contact with MHC, the proportion of CD127+CD132+ cells was decreased in both subpopulations of CD4+ memory cells. Upon in vitro cultures, IL-7 contributed to decreased expression of CD127, and increased expression of CD132 on CD4+Tcm and Tem. We have evaluated the CD4+Tcm and Tem populations by affinity of T cell receptor (TCR), using the level of CD5 expression. Т cells with high TCR affinity for self-antigens are known to have higher expression of CD5. In comparison to Tem, the Tcm contained more CD5high cells. In cultures, IL-7 promoted a high level of CD5 expression on Tcm, which was comparable to levels observed in peripheral blood cells. High CD5 expression on Tem was observed after stimulation with IL-7 in the in vitro inflammatory model. In the absence of contact with MHC, the number of CD5high cells decreased among CD4+Tem and Tcm. Thus, CD4+Tcm cells with high affinity for autologous antigens are probably dependent on the presence of homeostatic factors, in particular, IL-7, and contacts with antigen-presenting cells (APCs). Under conditions of inflammation, no changes were revealed in the mechanism of maintaining CD4+Tcm, in contrast to CD4+Tem. Being less dependent on IL-7 under normal conditions, CD4+CD5highTem are accumulated in the presence of IL-7 under in vitro inflammatory conditions.

It is known that ionizing radiation influences the expression of the genes that play a key role in the mechanisms of maintaining the stability of cellular homeostasis. As a rule, changes in the transcriptome of an exposed cell occur within the first 24 hours following radiation exposure. And it predetermines early response in the case of genome damage. Later on modulations in gene transcription activity are also possible and could result in a carcinogenic effect. However, in order to find the role of exogenous factors (ionizing radiation), it is also necessary to take into account the contribution of endogenous factors that are able to modify gene transcription activity. This is especially important for long after the onset of radiation exposure. Single nucleotide polymorphisms located in regulatory regions of the genes may belong to this group of factors. The objective of the current study was to analyze the influence of ionizing radiation on the transcription activity of the STAT3, GATA3, NFkB1, PADI4 genes, which regulate proliferation and differentiation of immune competent human cells; and to assess the potential influence of single nucleotide polymorphisms located in regulatory regions of the genes on the amount of mRNA. The study involved people who had been chronically exposed due to releases of radioactive waste into the Techa River. It was observed that 60 years after the onset of radiation exposure changes in the transcription activity of the NFkB1 and PADI4 genes were registered in people with cumulative doses to RBM within the range 78–3510 mGy. In people who had been chronically exposed, the effect of allelic variations in rs1053023, rs4143094, rs28362491, rs874881 on the level of mRNAs of the STAT3, GATA3, PADI4, NFkB1 genes has not been established.

The description of the method and algorithm for embedding a hidden message or a digital watermark into files of Microsoft Word electronic documents in .DOCX format based on two steganographic methods is given. A Microsoft Word electronic document in .DOCX format is used as a steganographic container. One of the methods uses the features of displaying a document by a word processor and the word processor allows the displacement of hidden characters, such as spaces, tabs and paragraphs, in the text relatively to the line of text. The second method uses the feature of the .DOCX format electronic text document that a document is an archive containing Open XML format files and media files, so specialized steganographic methods for XML files can be used for embedding a hidden message. In this case the quotes replacement method is used. The embedding of a hidden message by one of the methods is used for checking the integrity of the other message through the second method. Depending on the capacity of the steganographic container a method can be chosen to embed the message anda method to control the integrity of the message. The algorithm of the inverse steganographic transformation for extracting a message and confirming the integrity of an electronic document is considered. The application is developed to perform the embedding of a hidden message in an electronic text document depending on the capacity of the container. The possibility of using of some steganographic methods is analyzed with the aim of forming a multi-key steganographic system intended for a digital watermarking of an electronic document Microsoft Word format .DOCX.

Bronchial asthma is a chronic inflammatory disease of the respiratory tract. T-lymphocytes play a key role in pathogenesis of this allergic disease. The reduction in number of naïve T cells and the accumulation of memory T cells in bronchial asthma are accompanied by dysregulation of T lymphocyte function. In present study, we have investigated the contents of different T lymphocyte subpopulations in peripheral blood as well as in resting and PHA-stimulated cultures, along with their proliferative capacity in patients with bronchial asthma and healthy donors. The study included 10 patients with bronchial asthma (age 45.4±11.8 years). One-half of patients was in remission state, the others having been at the stage of clinical exacerbation. The group of donors was formed by healthy individuals matched by gender and age to the patients. Based on expression of cell surface markers CD45R0, CD62L and CD197 (CCR7), the CD4+ and CD8+T lymphocytes were divided into central (Tcm) and effector memory cells (Tem), naïve T lymphocytes (Tnaïve) and terminally differentiated effector cells (Temra) using flow cytometry technique. The proliferative activity of Tcm, Tem and Tnaïve was evaluated in response to PHA as a functional marker of T cells. We have found that the percentage of peripheral CD4+TemCD62L+ and CD8+TemCD62L+ cells in the patients with asthma exacerbation was significantly reduced, if compared to the donors. Following PHA stimulation, these differences in T cell subsets between the groups of patients and donors were not detectable. We performed a correlation analysis between the memory T cell contents and age of the subjects studied. It was shown that the relative amounts of CD4+ and CD8+ memory cells increased with age in asthmatics, but not in healthy donors. Analysis of mitogen-induced proliferation showed that Tcm and Tnaïve cells proliferated more actively than other subpopulations in both groups. Meanwhile, the proliferative activity of CD4+T lymphocytes and subsets of CD8+Tcm, CD4+Tcm and CD4+Tem62L was higher in the group of asthma patients in remission state than in the patients with exacerbating disease, and healthy donors. The revealed increase in the relative number of memory T cells with age suggests that these cells participate in development of bronchial asthma. Proliferative response of the studied subpopulations, which was comparable to the donor values, suggests a functional maintenance of memory T cells and naïve T lymphocytes in bronchial asthma. The increased proliferation of some T-cell subpopulations in asthmatics in remission suggests an activated state of memory T cells. The observed decrease in the number of CD4+TemCD62L+ and CD8+TemCD62L+ in patients with asthma exacerbation may be, by our opinion, associated with an active inflammatory process in the airways.

Objective. The purpose of this study was to evaluate the length of telomeres in the arms of individual chromosomes in patients with bronchial asthma (BA).Materials and methods. The study included patients with BA (n = 10, the mean age (44 ± 8.2) years) and healthy donors (n = 10, the mean age (44 ± 8.4) years). Metaphase spreads obtained from peripheral blood mononuclear cells were used. At the time of sampling BA patients received treatment at the Clinic of Immunopathology, Novosibirsk. BA was diagnosed by physicians according to GINA-2016. For measurement of telomere length on individual chromosome arms we used quantitative fluorescent in situ hybridization with a PNA-probe specific for telomeres. We used inverted DAPI banding for chromosome identification (according to ISCN-2013). For each individual 5 metaphase cells were analyzed. We applied the newly developed MeTeLen software to estimate the telomere repeats quantity (http:// www.bionet.nsc.ru/en/development/application-development/development-of-a-computer/metelen.html) in metaphase images. For enhanced image analysis compared with the previously developed programs, we included estimation of background signal and correction of defects of the optical system.Results. Comparing of telomere length show, that telomeres in the certain chromosome arms (4q, 5q, 9p, 10 q, 11p, 13p, 15q, 18q, 19q) in BA are significantly shorter than in corresponding group of donors (p < 0.05, Mann – Whitney U-test). For both studied groups we also evaluated telomere sequences shortened and elongated relative to the average telomere length in the group (p < 0.05, Wilcoxon-signed-runk test). The following differences and similarities between the telomere profiles of patients and donors were determined: the telomere sequences 4p, 6q, 8p were elongated and 2q, 9q, 11p, 15q were shortened relative to the average telomere length in BA patients. Moreover, this telomere sequences did not differ from the average telomere length in the group of donors. At the same time, the telomere sequences 12p, 16p, 17p, 19p were significantly shorter, and 3p was longer than the average telomere length in both groups.Conclusions. We guess, that the observed significant shortening of telomere length on individual chromosome arms in BA, as compared to donors, is relevant in pathogenesis of this disorder. The revealed features of telomere profile of patients with BA may be a result of different telomere length maintenance mechanisms and may influence to the development of asthma that needs further study.

The aim of the study was to investigate the features of T-lymphocyte apoptosis induced by components of autologous apoptotic cultures in vitro in norm and rheumatoid arthritis in the context of «cellular neighborhood».Materials and methods. Subjects of the study were blood samples of patients with rheumatoid arthritis (RA) and healthy women of comparable age. Developed protocol allowed to differentially evaluate the parameters of proliferation, early and late stages of apoptosis in the «primary» (CFSE-) and «secondary» (CFSE+) induced apoptotic T-lymphocyte cultures. It was estimated the effect of cellular and humoral components of unstimulated, anti-CD3- and dexamethasone-stimulated cells under the conditions of overcrowding and depleted culture media on autologous lymphocytes, cultured under physiological conditions, in norm and RA.Results. Comparative qualitative analysis revealed the features of the processes of T-lymphocyte apoptosis in norm and pathology. Also, the parameters of early and late stages of apoptosis of a «primary» induced culture and «secondary» induced cells after transferring the cellular and humoral components of apoptotic cultures did not differ significantly either initially or during culturing in both investigated groups. But it was a significant increase in the amount of living T-cells in «primary»-induced unstimulated and dexamethasone-stimulated RA patients’ cultures compared to similar donors’ cultures.Conclusion. There was no difference between stimulated with anti-CD3 antibodies cells and the «secondary» induced cultures. Taking into account the absence of significant differences in the parameters of activation apoptosis, the increased number of living cells in RA patients’ cultures relative to donors’ is evidence of contribution of non-autonomous apoptosis effects to cellular homeostasis in RA.

Aim. The aim of this study was the investigation of the influence of humoral factors of homeostatic proliferation IL-7 and IL-15 on T-regulatory cells in healthy donors.Materials and methods. The study included 15 conditionally healthy donors. Phenotyping and evaluation of expression changes of transcription factor FoxP3 and the main functional molecules on T-regulatory cells such as PD-L1, CTLA-4 and HLA-DR during cultivation under IL-7, IL-15 and anti-CD3 stimulation combined with IL-2 were performed by flow cytometry. Also, we estimated proliferation intensity of T-regulatory cells in the course of cultivation.Results. We revealed that humoral factors of homeostatic proliferation can effectively support a pool of T-regulatory cells during cultivation by number and phenotype and can maintain expression of important molecules such as PD-L1 and HLA-DR on regulatory T-cell surface. In addition, our study showed that IL-7 and IL-15 can cause relatively low T-regulatory cells proliferation in comparison to CD4+- lymphocytes.Conclusion. The observed ability of homeostatic proliferation factors to maintain T-regulatory cells pool presumably can play an important role in lymphopenic conditions when the number of effector cells is decreased and the insufficiency of interleukin IL-2 is observed, which plays a primary role in the homeostasis of T-regulatory cells in normal conditions.

Defects in programmed death of peripheral blood lymphocytes may result into autotolerance and chronic persistence of inflammatory reaction in rheumatoid arthritis. According to recent data, apoptosis is not an autonomous process, i.e., an excessive accumulation of cells in conjunction with a depleted medium leads to cellular excess, thus causing effect of cells already committed to apoptosis upon surrounding cells, by morphogenetic damage due to mechanical forces. Such effects are called compensatory proliferation or apoptosis-induced proliferation. Extracellular vesicles (ectosomes and apoptotic bodies) also have a negative effect on cultured cells, triggering their programmed apoptotic death. In turn, apoptosis can also be controlled by neighboring cells in non-autonomous manner.We conducted studies that allowed us to optimize methods aimed at the initiation of apoptotic cell death and to investigate the effects of apoptotic environment upon autologous cells under physiological conditions. The selected conditions in combination with a fluorescent labeling of lymphocytes and subsequent separate flow cytometric analysis allowed us to evaluate parameters of early apoptosis in subpopulations of peripheral blood T-lymphocytes in rheumatoid arthritis. In vitro studies of cells from the patients with rheumatoid arthritis allowed us to reveal a pronounced readiness of primary (CFSE) and secondary (CFSE+)-induced T lymphocytes for early apoptosis after stimulation with anti-CD3 antibodies. It was observed both against initial level of apoptosis, and when compared to cells induced for apoptosis. The obtained data suggest that stimulation of T lymphocytes with antibodies against CD3, and, as a result, an in vitro rise in cell proliferation rate leading to increased levels of early apoptosis not only among the cells directly receiving a proliferative stimulus, but also to increased effect of cellular and humoral components from anti-CD3-stimulated cultures upon normally proliferating lymphocytes.The transfer of an autologous apoptotic “aCD3” and dexamethasone-stimulated cultures, which were initially induced under conditions of cell overcrowding and medium exhaustion, was shown to activate the process of early apoptosis among normally proliferating cells. Glucocorticoids are known to serve as agents of cell death induced by activation. At pharmacological concentrations, glucocorticoids and their synthetic analogues stimulate endonucleases in activated lymphocytes. These enzymes destroy DNA in the internucleosomal regions thus resulting into cell apoptosis. The results obtained in present study suggest an opportunity of an in vitro early-stage apoptosis induction in T lymphocytes from the patients with rheumatoid arthritis, by means of cells subjected to activation-induced apoptosis.

Abstract. Functional activity of thymus in normal donors and patients with various clinical forms of bronchial asthma was evaluated by means of assessing TREC-positive T-cell numbers. In spite of well-known alterations in parameters of T-cell homeostasis, TREC levels in CD4+and CD8+ lymphocytes from the patients with asthma did not differ significantly from those in donors. There were also no significant differences in TREC-containing cell numbers, dependent on clinical types of bronchial asthma and IgE levels. However, the patients with a recent disease onset (duration of several days to one year) showed increased TREC positivity in CD4+ and CD8+ lymphocytes, as compared with donors and those patients who underwent basal therapy with glucocorticoids, and during long-term disorder (> 1 year). Enhanced thymopoiesis at initial stages of the disease may be associated with thymic stromal lymphopoietin which contributes to development of bronchial asthma, and normally participates in the process of T cell production.

For the first time specific Ig was obtained from blood plasma of donors immunized by combined anthrax vaccine. This Ig conforms to quality standards for intravenous immunoglobulins and includes antibodies to Bacillus anthracis protective antigen in titer 1:1600 - 1:3200. Therapeutic and prophylactic efficacy of experimental-production series of preparation was demonstrated in experiments with animals.

Aim. The article presents data on the evaluation of the clinical efficacy of using a conjugated pneumococcal vaccine in patients with COPD in combination with 2-type diabetes during a 3-year follow - up period. Materials and methods. The study included 309 patients with an isolated course of COPD and in combination with diabetes. The main parameters for evaluating the effectiveness were: the dynamics of clinical symptoms - shortness of breath on the mMRC scale, CAT test, FEV1, the number of exacerbations, hospitalizations, the number of pneumonia, the state of carbohydrate metabolism (HbA1c) and the lipid profile. For vaccine prevention 13-valent conjugated pneumococcal vaccine Prevenar-13 was used. Results and conclusions. The use of PСV13 helps to reduce the severity of respiratory symptoms, reduce the number and duration of exacerbations of COPD, reduce the number of pneumonia. Both in isolated course of COPD and in combination with diabetes. Vaccination PCV13 in patients with comorbid course contributes to the compensation of carbohydrate metabolism and improve the lipid profile.

The paper describes a case of primary diagnosis of functioning patent ductus arteriosus in a 75-year-old female patient.

The aim of the study was to evaluate the possibilities of left ventricular hypertrophy (LVH) detection with the use of a complex including a system of remote 12-lead electrocardiography (ECG) analysis, orthogonal lead ECG and vectorcardiography (VCG), during the medical examination of the population of Uryupinsk district of the Volgograd region. The work was organized in two rural district hospitals of Uryupinsk district. 180 persons were randomly selected out of the total data bank. Of these, according to ECG and VCG, 54 persons (30%) were considered to be practically healthy, and in 63 cases (35%) automatical analisys of ECG or VCG revealed signs of LVH. In 32 cases (50.7%) left ventricular hypertrophy was revealed for the first time. The most often finding was the increase of the parameters of synthesized orthogonal ECG and VCG: the sum of Rx+Sz and maximal QRS vector (89%, 95%). About 10% of the patients had high blood pressure without electrocardiographic signs of LVH. Introduction of the remote ECG analisys to the work of medical institutions of Uryupinsk district allowed to increase LVH detection.