The role of autophagy in the regulation of neuroinflammation in acute ischemic stroke (review of literature).

Postischemic neuroinflammation is a critical pathophysiological process within the entire scheme of cerebral ischemia, covering early damage and the period of tissue repair. It is characterized by microglial and astroglial activation with increased expression of inflammatory mediators and is accompanied by impaired innate and adaptive immune responses. In acute ischemic stroke (IS), neuroinflammation is caused by the response of resident immune cells of microglia and peripheral immunocompetent cells infiltrating the brain tissue, which penetrate the blood-brain barrier (BBB) into the lesion. Recent studies have shown the important role of the NLRP3-mediated inflammation in the death of neurons and glial cells in acute IS. The review presents the main mechanisms of activation of NLRP3-mediated inflammation in acute IS, leading to the caspase-1 formation and the IL-1ß and IL-18 release, which are involved in the initiation and progression of inflammation in the brain parenchyma. The literature data on the role of autophagy in the inhibition of postischemic neuroinflammation are summarized. Autophagy can suppress neuroinflammation through a wide range of the autophagy - related proteins. The role of autophagy as a negative regulator of NLRP3-mediated inflammation in acute IS is analyzed. Data on the participation of autophagic proteins Beclin-1, LC3, and p62 in the suppression of NLRP3 inflammation due to the induction of basic mitophagy are presented. Prospects for modulating autophagy aimed at suppressing postischemic neuroinflammation, including the inhibition of NLRP3-inflammasome, have been noted. The review was based on sources from international and national data bases: Scopus, Web of Science, Springer, RINC.

[The phenomenon of pseudotumorous cross immunohistochemical reactivity of glia in progressive multifocal leukoencephalopathy]. / Fenomen psevdotumoroznoi perekrestnoi immunogistokhimicheskoi reaktivnosti glii pri progressiruyushchei mul'tifokal'noi leikoentsefalopatii.

Progressive multifocal leukoencephalopathy (PML) is a subacute demyelinating brain damage caused by infection of oligodendrocytes and astrocytes with the lytic JC virus on the background of immunosuppression. A case report of PML with a tumor-like course is presented. Morphological diagnostics revealed non-specific staining of antibodies to Ki-67, p53, IDH1, NF and Vim in the nuclei of gliocytes affected by the JC virus. Histological examination and microscopic evaluation of the changes in the brain for the diagnosis of PML is a priority. The recommended intravital biopsy does not always help in clear verification of PML due to the limited volume of tissue fragments presented for research. For the correct interpretation of changes during an intravital pathological examination and verification of PML, it is important to take material during a stereotaxic biopsy, not only from the center, but from the edges and perifocal zone of the altered tissues for the possibility of a spatial histological assessment of the pathological process.

Computer-Aided Detection of Respiratory Sounds in Bronchial Asthma Patients Based on Machine Learning Method.

The aim of the study is to develop a method for detection of pathological respiratory sound, caused by bronchial asthma, with the aid of machine learning techniques. Materials and Methods: To build and train neural networks, we used the records of respiratory sounds of bronchial asthma patients at different stages of the disease (n=951) aged from several months to 47 years old and healthy volunteers (n=167). The sounds were recorded with calm breathing at four points: at the oral cavity, above the trachea, on the chest (second intercostal space on the right side), and at a point on the back. Results: The method developed for computer-aided detection of respiratory sounds allows to diagnose sounds typical for bronchial asthma in 89.4% of cases with 89.3% sensitivity and 86.0% specificity regardless of sex and age of the patients, stage of the disease, and the point of sound recording.

Clinical implication of detecting sensitization to iodinated radiocontrast media in the basophil activation test by flow cytometry.

The use of iodinated radiocontrast media is necessary for visualization. A number of patients have adverse effects of various nature and severity when these drugs are administered. Routine allergy tests do not provide adequate diagnosis of reactions to drugs in this group. The aim of this work is to assess the capabilities of the basophil activation test to confirm sensitization to non-ionic iodinated radiocontrast media, as well as to select a safe alternative drug in patients with a burdened history. Basophil activation test by flow cytometry was performed in 184 patients The Nikiforov Russian Centre of Emergency and Radiation Medicine¼ EMERCOM of Russia and 32 volunteers using ultravist, omnipack, and optiray. The presence of sensitization was assessed based on the basophil activation index, as well as spontaneous and anti-IgE antibody-induced activation of basophils and the population of T-lymphocytes type 2 immune response. The volunteers showed no sensitization to iodinated radiocontrast media. In patients with a medium degree of hypersensitivity reaction in vivo, in vitro sensitization to drugs was detected 4 times more often than in patients with a mild degree (51% versus 13.5%). In patients with systemic reactions to the administration of a known drug, in vitro sensitization was confirmed in 86% of cases, while the frequency of detection of sensitization to drugs did not differ. Spontaneous activation of basophils in patients and type 2 T-lymphocytes were 2 times higher than in volunteers. Patients were more likely to have low (less than 30%) activation of basophils for anti-IgE antibodies. The specificity of the basophil activation test with iodinated radiocontrast media was 100% with a sensitivity of 94.1%. Most patients were able to select a non-sensitizing contrast. Inclusion in the algorithm of spontaneous and anti-IgE antibody-induced activation of basophils and a population of T-lymphocytes type 2 immune response will allow the doctor to carry out a personalized approach to the management of patients with a burdened history.

[Apoptosis and autophagy in the pathogenesis of acute ischemic stroke (review of literature).] / Апоптоз и аутофагия в патогенезе острого ишемического инсульта (обзор литературы).

According to the World Health Organization, about 5 million people die every year from cerebrovascular disease. At the same time, the proportion of cerebral infarction, or ischemic stroke (IS), among forms of acute cerebrovascular accident reaches 80-85%. Despite the active study of biochemical and morphological changes leading to acute cerebrovascular ischemia, the problem of early diagnosis, prevention, as well as predicting the outcome of this disease is still relevant. There is no doubt that the interruption of the ischemic cascade at earlier stages can be accompanied by a greater effect of treatment. A timely and effective pharmacological intervention requires a clear understanding of the pathochemical and biological processes underlying acute ischemia at the molecular level. High mortality and disability accompanying acute IS, dictate the need to create new diagnostic and prognosis algorithms both in the acute period of IS, and in the recovery period. According to some authors, elucidation of the pathways that underlie the pathogenetic mechanisms acting in the penumbra are of great clinical interest for the development of new diagnostic and therapeutic strategies. Studying the mechanisms of apoptosis and autophagy of neurons in the dynamics of the acute period of IS, modulation of the autophagy process in the penumbra zone can contribute to the development of new methods for the diagnosis and treatment of acute IS. The review presents the results of the latest experimental studies on the role of apoptosis and autophagy in the development of acute cerebral ischemia and attempts to modulate these processes in order to influence the ischemic cascade. The review was based on sources from such international and national data bases as Scopus, Web of Science, Springer, RINC.

[Prevention and treatment of degenerative changes in ocular surface epithelium in patients with dry eye syndrome]. / Profilaktika i lechenie degenerativnykh izmenenii épiteliia glaznoi poverkhnosti pri sindrome 'sukhogo glaza'.

In current understanding of the pathogenesis of xerotic changes of the ocular surface in patients with dry eye syndrome (DES), great importance is given to hyperosmolarity of the tear film, inflammatory process and oxidative stress. By now, no convincing data has been obtained on whether the oxidative stress is primary in relation to the inflammatory process in the ocular surface tissues, or if it is a complication. Furthermore, in the complex treatment of patients with DES, antioxidant therapy has so far received little attention. This problem can be solved with the 'artificial tears' preparation containing cyanocobalamin (vitamin B12), which antioxidant, anti-inflammatory and clinical effectiveness has been convincingly proved in a number of experimental and clinical studies of recent years. Improving the methods of vital diagnostics of xerosis of the ocular surface also enables detection of subclinical changes in epithelium of the cornea and conjunctiva, thus allowing timely prescription of metabolic therapy. One of the promising directions of such treatment is the use of 5% dexpanthenol, which stimulates the processes of reparative regeneration and possesses an anti-inflammatory effect.

[FROM THEORY TO PRACTICE. THE ROLE OF QUALITY CONTROL OF ANALYTICAL STAGE OF STUDIES IN INCREASING CLINICAL INFORMATIVENESS OF LABORATORY TESTS].

Nowadays, improving accuracy and clinical informativeness of results of studies is one of main directions of development of laboratory. The article presents originally developed algorithm of evaluation of quality of immunochemical studies in part related to analytical stage. The case of application in practice of the results of such evaluation is presented.

[Multiple sclerosis vaccine: possibilities of the use of dendritic cells immunomodulatory potential].

For the first time we carried out a clinical assessment of the safety, tolerability and clinical efficacy course of repeated administration of experimental modified autologous vaccine interleykin (IL-10) dendritic cells in two patients with secondary-progressive multiple sclerosis patient and one with relapsing-remitting multiple sclerosis. In the course of treatment, we carried out clinical and immunological monitoring. It was found out that intradermal dose of 3 x 106 cells applied to spinal area 6-12. times did not cause any serious side effects. After the treatment with dendritic cells, the following results were observed: 1) a significant positive clinical effect in patients with secondary-progressive multiple sclerosis exacerbations; 2) moderate positive clinical effect in patients with relapsing-remitting multiple sclerosis, in a state of remission; 3) a complete absence of any clinical results in patients with secondary-progressive multiple sclerosis without exacerbations. The immune response was characterized by a significant absolute and relative increase of serum T-regulatory cells. Discovered distinct anti-inflammatory properties of dendritic cell therapy allow us to consider it as a promising area of personalized treatment based on an individual vaccination against multiple sclerosis.

[The state of immunity in the liquidators of consequences of the Chernobyl accident with cardio-vascular diseases].

The purpose of the work was research and functional reserves immunity in participants in the Chernobyl with cardiovascular disease. A Clinical Lab 49 men aged 44 to 52 years with diseases of the cardiovascular system, participated in the Chernobyl nuclear power plant in 1986 to 1988. As control used data 33 patients with similar pathology, and 16 healthy men. Researched the total number of white blood cells, lymphocytes and transfusions, the absolute number and relative CD4+ and CD8+, CD 16, CD20+, CD95+ lymphocytes peripheral blood, number mononuclear, synthesizing IL-1beta, TNF-alpha, INF-alpha, IL-4, the content of lipids and proteins, the activity of alkaline phosphatase and mieloperoksidazy in neutrophils. To assess the functional reserve immune system blood samples studied people subjected to radiation doses 0.25, 50, 1.0 Gy of in vitro and studied the reaction cytochemical indicators neutrophils before and after the radiative forcing. The liquidators not detected significant changes in the absolute number of leucocytes, but compared with control groups noted significant reduction in the absolute number of CD8+ and CD20+ lymphocytes, increased the number of cells, expression of FAS-antigen, change the number of mononuclear spontaneously synthesizing and produce cytokines, decreased maintenance of cation proteins in neutrophils. Radiation samples peripheral blood liquidators caused the same reaction cytochemical indicators of neutrophils and control groups, the compensatory and adaptive nature of the changes in the immune system, developed in response to complex factors radiation accident.

Induction of the estrogen effect-switching phenomenon by ethanol and its correction.

Female rats (aged three months at the start of the study) were kept for four months on drinking water (group 1) or 5% ethanol. Rats drinking ethanol were additionally divided into six groups (groups 2-7). During the next two months of the experiment, animals of group 2 continued to drink only ethanol, while rats of group 3 additionally received N-acetylcysteine, those of group 4 received ascorbic acid (vitamin C) and alpha-tocopherol (vitamin E), those of group 5 received melatonin, those of group 6 received carnosine, and those of group 7 swam in the so-called training regime for five days a week. All animals underwent bilateral ovariectomy 2.5 weeks before experiments ended, and were given daily i.m. estradiol (2 microg) during the 11 days before the last experimental day. After treatments, blood estradiol and cholesterol levels were measured, along with progesterone receptor levels, peroxidase activity, the index of proliferation, the proportions of cells in the S and G2/M phases, the thickness of the endometrium, and the extent of DNA damage (using the "comet" technique) in uterine tissue. Liver estradiol 2-hydroxylase activity was also measured. The results led to the conclusion that drinking of 5% ethanol in combination with administration of estrogens was accompanied by induction of genotoxic (G) changes in the uterus and that this was prevented by administration of N-acetylcysteine and melatonin. The combination of vitamins C and E increased some of the manifestations of the hormonal (H) effect of estrogens (uterine weight and induction of progesterone receptors), but weakened others (the index of proliferation). As a result, the combination of N-acetylcysteine and optimum doses of ascorbic acid and alpha-tocopherol can be recommended for preventing the estrogen effect-switching phenomenon (increases in the G component on the background of weakening of the H component), which is seen particularly in patients consuming excessive amounts of alcohol, increasing the risk that the genotoxic version of hormonal cancerogenesis will develop.

Modification of uterotropic effect of estrogens by whole-body gamma-irradiation.

The effect of gamma-irradiation on the realization of the effects of estrogens was studied on rats treated with N-acetylcysteine, vitamins C and E, melatonin, and carnosine or subjected to forced swimming in a training mode. Irradiation (0.2 Gy) in combination with estrogens and without correction therapy induced genotoxic changes in the uterus, while irradiation in a higher dose (2 Gy) predominantly potentiated the hormonal effect of estrogens. Correction of the revealed abnormalities was achieved mainly with carnosine. The peculiarities of "estrogen toggle (re-targeting) effect" under the effect of gamma-irradiation and its elimination differed from those induced by ethanol intake or tobacco smoking, which is important for understanding the mechanisms of hormone-induced carcinogenesis.

[Immunogenetic cytokine restriction in multiple sclerosis]. / Immunogeneticheskaia resstriktsiia tsitokinov pri rasseiannom skleroze.

Sixty eight patients with verified multiple sclerosis (MS) (mean EDSS score 3.1 +/- 1.0) and 50 healthy donors have been investigated. Thirty five patients had relapsing-remitting, 25--secondary progressive, 8--primary progressive course. The remission was in 38, decompensation--in 20, relapse--in 10 patients. Lymphocyte subpopulations were investigated using monoclonal antibodies (Moscow) to the following antigens: CD3 (T-lymphocytes), CD4 (T-helpers), CD8 (T-supressors), CD20 (8-lymphocytes), CD25 (IL-2 receptor), CD16 (natural killers), CD95 (activated cells ready to apoptosis). Cytokines and tumor necrosis factor-alpha (TNF-alpha) levels were measured using ELISA test. HLA antigens were investigated by standard lymphocytotoxic test. In MS we found a fall of CD3, CD4, CD8, CD20 and CD16, but an increase of CD4/CD8, CD95, CD25. The CD95 level correlated with CD4, CD4/CD8 and CD16. In MS spontaneous IL-2, IL-6, IL-8 and TNF-alpha production was raised and stimulated IL-6 and IL-8 secretion was reduced. IL-4, IL-6, IL-8, TNF-alpha and IL-1 beta serum production in vivo was elevated. We found an increase of CD3, CD4, CD16, CD25, but a decrease of IL-1 (p < 0.01) spontaneous production and IL-6, IL-8, TNF-a stimulated secretion in DR2(+) MS patients, comparing to DR2(-) patients and controls. In DR2(-) patients as compared to DR2(+) patients and controls, all lymphocyte subpopulations levels, especially CD8 (p < 0.001) one, were decreased, but spontaneous IL-8 (p < 0.01) production was increased. The data obtained indicate lymphocyte apoptosis activation, targeting promoted lymphocyte destruction, and suggest T helper type-1 reaction prevalence in MS.

[Induction by ethanol of 'phenomenon of switching estrogen effect' and its correction]. / Induktsiia étanolom "fenomena perekliucheniia éffekta éstrogenov" i ego korrektsiia.

Drinking of 5% ethanol in combination with estradiol induced genotoxic (G) changes in the rat uterine tissue. The changes could be prevented by N-acetylcysteine or melatonin. The data obtained suggest that ascorbic acid and alpha-tocopherol may be recommended in combination with N-acetylcysteine for repair of estrogen effect switching phenomenon.

[Specific features of diphtheria vaccinal process in children with rheumatic diseases]. / Osobennosti techeniia difteriinogo vaktsinal'nogo protsessa u detei s revmaticheskimi zabolevaniiami.

Clinical and immunological examination of 55 children aged 6-15 years with rheumatic diseases, immunized against diphtheria, was carried out. All children were immunized at the stage of clinical and laboratory remission and in some cases while undergoing a prolonged course of cytostatic therapy or therapy with nonsteroid anti-inflammatory remedies. This examination demonstrated that in the overwhelming majority of children with rheumatic diseases the diphtheria vaccinal process took an asymptomatic course and had no influence on the course of the main disease. Specific features, characteristic of the immune status of this group of children, were established. In the course of the vaccinal process the restoration of the initially inhibited characteristics (the production of TNF-alpha and IL-2) to normal values were shown to occur, which was indicative of the fact that the reserve capacities of immunocompetent cells were retained in these patients. This study also revealed that immunization of children with rheumatic diseases with adsorbed DT and D toxoids with reduced antigen content was not excessive antigenic stimulation for such children, as it did not lead to immunopathological shifts, but induced transient phase changes in immunological characteristics, similar to those in healthy children. Protective levels of antibodies to diphtheria were shown to retain for a long time with considerable prolongation of intervals between booster injections. The simultaneous course of immunosuppressive maintenance therapy in the average dosage used for the corresponding age group did not inhibit the production of protective antibodies.

[Multiple sclerosis in Northern-West region of Russia: results of HLA-typing]. / Rasseiannyi skleroz v Severo-Zapadnom regione Rossii: rezul'taty HLA-tipirovaniia.

Distribution of antigens of A, B, DR loci of HLA system in standard lymphocytotoxic test was studied in 59 patients with a significant diagnosis of multiple sclerosis (MS) and in 138 healthy donors. In the patients elevated frequency of the next antigens was found as compared with the controls: A10 (37%; chi 2 = 6.31; p < 0.05; relative risk--RR = 2.34), B7 (37%; chi 2 = 4.62; p < 0.05; RR = 2.05), B13 (29%; chi 2 = 10.86; p < 0.01; RR = 3.59), B35 (17%; chi 2 = 4.27; p < 0.05; RR = 2.61), DR2 (68%; chi 2 = 11.61; p < 0.001; RR = 2.99), as well as DR6 (5%; chi 2 = 3.95; p < 0.05; RR = 7.34) and also DRw52 (24%; chi 2 = 27.49; p < 0.001; RR = 21.16). The highest value of etiologic fraction was found for DR2 antigen. Analysis of intralocus and extralocus combinations of antigens in MS revealed that significantly elevated frequency had only one combination--B7DR2 (25.4%; chi 2 = 9.77; p < 0.01; RR = 3.58), relative risk was higher for this combination than for each individual antigen separately: B7 (RR = 2.05), DR2 (RR = 2.99). Significant negative associations with a possible protective effect of separate alleles were established in MS for antigens HLA A2 (34%; chi 2 = 5.55; p < 0.05; RR = 0.47), A11 (7%; chi 2 = 4.66; p < 0.05; RR = 0.31), A30 (0%; chi 2 = 4.50, p < 0.05; RR = 0.01), B18 (8%; chi 2 = 4.55; p < 0.05; RR = 0.35), DR5 (59%; chi 2 = 10.17; p < 0.01; RR = 0.36). The most significant was a decrease of the frequency of DR5 antigen (p < 0.01). Patients with the recurrent course had prevailed antigens A11, B21, B35 and decreased frequencies of antigens A9, B13, DR7. However, only the difference in the frequency of DR7 (16% in remitting and 57% in progredient course, chi 2 = 10.02; p < 0.001; RR = 0.14) was significant.

Effect of ethanol on uterotropic action of estrogens.

Female rats aging 3 months at the beginning of experiments received 5 or 15% ethanol and then were subjected to bilateral ovariectomy 2 weeks before end of the experiment. During the last 11 days they were daily injected intramuscularly with 2 microg estradiol. Drinking of 5% ethanol combined with injections of estrogens induced DNA damage in the uterus detected by comet assay and abolished induction of progesterone receptors, changes in peroxidase activity, proliferation index, endometrium thickness, and other indices reflecting the hormonal effect of estradiol on the uterus. Drinking of 15% ethanol was accompanied by an increase in DNA-damaging effects of estrogens and a decrease in their hormonal uterotropic effects. It is concluded that unlike tobacco smoking, drinking of moderate ethanol concentrations modifies primarily genotoxic, but not the hormonal effect of estrogens.

[Changes in the production of interleukin-1 beta, tumor necrosis factor-alpha and interleukin-2 in relation to the immunity in patients with pulmonary tuberculosis]. / Izmeneniia v produktsii interleikina-1beta, faktora nekroza opukholi alfa i interleukina-2 v zavisimosti ot sostoianiia immuniteta u bol'nykh tuberkulezom legkikh.

The specific features of production of IL-1 beta, TNF-alpha, IL-2 were studied in 74 patients with various forms of tuberculosis by taking into account the magnitude of an immunological response. Tuberculin, phytohemagglutinin, prodigiosine were used as inducers of the synthesis of cytokines. Heterodirection was found in the changes of elaboration of cytokines in similar immunological disorders in persons with different clinical forms of tuberculosis. Examination of patients with infiltrative tuberculosis indicated that the increased synthesis of TNF-alpha and IL-2 was to a greater extent associated with the activation of cell-mediated immunity and that of IL-1 beta with its inhibition. The relationships found between the production of cytokines and IgA and IgM levels are suggestive of their involvement in the regulation of immunoglobulin synthesis. Cytokine spectral alterations are associated with the changes in specific lymphocytic populations. In patients with infiltrative tuberculosis, the production of TNF-alpha and IL-1 beta was directly related to the proportion of CD4+ and CDS8+ and that of IL-2 is associated with the proportion of CD25+ and CD20+ and with the count of lymphocytes. At tuberculin stimulation of mononuclear cells, it is expedient to bear in mind the detection rate of cytokines and the level of their production. It was shown that the measurement of IL-1 beta, TNF-alpha, IL-2 may be used in the treatment of tuberculosis to assess the patients' immunological response and in the choice of immunomodulating therapy.