[Antibiotic resistance of Helicobacter pylori in the European part of the Russian Federation: first results].

AIM: Determine the primary antibiotic resistance of Helicobacter pylori (H. pylori) strains isolated from patients living in the European part of the Russian Federation. MATERIALS AND METHODS: As part of a clinical laboratory study, from 2015 to 2018, 27 gastrobiopsy samples obtained from H. pylori-infected patients were analyzed. H. pylori infection was verified using a rapid urease test or a 13C-urea breath test. The values of the minimum inhibitory concentration (MIC) of antibiotics were determined by the diffusion method using E-test strips (BioMerieux, France) according to the recommendations of the manufacturer. The sensitivity of the isolates was determined for 6 antibacterial drugs (amoxicillin, clarithromycin, metronidazole, levofloxacin, tetracycline, rifampicin). RESULTS: According to the data obtained, resistance to amoxicillin was 0%, clarithromycin 11.1%, metronidazole 59.3%, levofloxacin 3.7%, tetracycline 0%, and rifampicin 14.8%. Dual resistance to clarithromycin and metronidazole was recorded in two isolates (7.4%). CONCLUSION: Thus, the first results of the evaluation of H. pylori antibiotic resistance in the European part of the Russian Federation indicate a low resistance of the microorganism to clarithromycin and quite high to metronidazole.

[Current issues in the rehabilitation of stroke patients against the background of a new coronavirus infection (COVID-19). Resolution of the Council of Experts]. / Aktual'nye voprosy reabilitatsii patsientov s insul'tom na fone novoi koronavirusnoi infektsii (COVID-19). Rezolyutsiya Soveta ekspertov.

The pandemic of the new coronavirus infection caused by SARS-CoV-2 has forced to reconsider the methods of rehabilitation of patients with emergency conditions, including neurology. The Council of Experts of Neurologists and Rehabilitation Therapists gathered to develop unified approaches to manage stroke patients based on a discussion of practical experience and, taking into account the scientific information on COVID-19 that was available by the time of the meeting. Stroke is a serious disabling condition that requires maximum rehabilitation efforts at all stages of medical care. In the context of the SARS-CoV-2 coronavirus epidemic, the process of medical rehabilitation and the routing of patients with stroke is undergoing major changes. Combining COVID-19 and stroke requires new approaches to rehabilitation and patient management. During the meeting, a resolution was developed in which the experts formulated the tactics of medical rehabilitation of patients with stroke and COVID-19 at the first and second stages. The arguments of the importance and practicability of carrying out measures of medical rehabilitation at the third stage is given and the need to continue consultations on the indicated topic is revealed.

[Influence of the Donor of Hydrogen Sulfide GYY4137 on the Activation of Human Neutrophils by E. coli Lipopolysaccharides].

Lipopolysaccharides (LPS), components of the cell wall of gram-negative bacteria, activate neutrophils that trigger pathological processes, including gram-negative sepsis. LPS inhibit spontaneous apoptosis of neutrophils that leads to inflammation. In this work we tested the action of H2S donor (GYY4137) on the activation of human neutrophils by E. coli LPS. We estimated the changes in redox status (ROS level, intracellularglutathione, NO), apoptosis and mitochondrial potential of neutrophils under the LPS action in the presence and absence of GYY4137. GYY4137 reduces the ROS level, slightly reduces GSH, does not influence the NO level and has no apoptogenic effect. LPS induce the increasing of ROS level and inhibit spontaneous apoptosis of neutrophils. We found that GYY4137 prevents the growth of ROS caused by LPS and leads to a reduction of LPS-induced inhibition of neutrophil apoptosis. Thus the mechanism of GYY4137 protection against inflammation, triggered by bacterial infection, is concerned with the neutralization of LPS effect on neutrophils.

[Results of a randomized double blind multicenter placebo-controlled, in parallel groups trial of the efficacy and safety of prolonged sequential therapy with mexidol in the acute and early recovery stages of hemispheric ischemic stroke (EPICA)]. / Rezul'taty randomizirovannogo dvoinogo slepogo mul'titsentrovogo platsebo-kontroliruemogo v parallel'nykh gruppakh issledovaniia éffektivnosti i bezopasnosti meksidola pri dlitel'noi posledovatel'noi terapii u patsientov v ostrom i rannem vosstanovitel'nom periodakh polusharnogo ishemicheskogo insul'ta (ÉPIKA).

AIM: To evaluate the efficacy and safety of prolonged sequential therapy with mexidol in the acute and early recovery stages of hemispheric ischemic stroke (IS). MATERIAL AND METHODS: A randomized double blind multicenter placebo-controlled, in parallel groups trial included 151 patients (62 men and 89 women) with hemispheric IS. Using a method of simple randomization, 150 patients (62 men and 88 women), aged 40-79 years, were randomized into two groups. Patients of Group I were treated with mexidol: 500 mg/day IV infusion for 10 days, followed by 125 mg tid (375 mg/day) PO for 8 weeks. Patients of Group II received the placebo according to the same scheme. The total duration of patients' participation in trial ranged from 67 to 71 days. RESULTS: By the end of treatment, the mean score on the modified Rankin scale (mRS) was lower in Group I compared to Group II (p=0.04). In Group I, the decrease in mRS mean score (Visit 1-5) was more prominent (p=0.023), percentage of patients with 0-2 scores by mRS scale (Visit 5) was higher (p=0.039), mean NIHSS score lower (p=0.035) in Visit 5 compared to group II. By the end of treatment, the decrease in mean NIHSS score in patients with diabetes mellitus was more prominent in Group I in comparison with Group II (p=0.038). In Group I, the dynamic of improvement of quality of life was more prominent and started from Visit 2 in general population and subpopulation of patients with diabetes mellitus. The share of patients with no problems with movement in space was higher in Group I (p=0.022). There were no statistically significant differences in frequency of side effects in patients of both groups. CONCLUSION: It is recommended to include mexidol in therapy of patients with IS in the acute and early rehabilitation stages.

[Preclinical studies of drugs on animal stroke models]. / Doklinicheskie issledovaniia lekarstvennykh sredstv na biologicheskikh modeliakh insul'ta.

Preclinical studies are studies using experimental models of stroke in animals as well as on neurons, cell neuronal cultures and surviving brain slices. They directed both towards testing the efficacy and evaluation of the mechanisms of action of drugs, and the study of the mechanisms of ischemic damage to search for new targets for stroke treatment. This article shows the basic principles of the organization and planning of animal models of ischemic stroke. Modeling of cerebral ischemia on the different models and animal species, the modern principles of assessment of brain damage are considered as well.

[BEHAVIOR, MEMORY AND IMMUNOLOGICAL STATUS IN MICE MODEL OF DESYNCHRONOSIS].

Interstrain differences in behavior and parameters of the immune system of CBA and C57BL/6 mice with round the clock coverage (KO) were investigated. Open field, light/dark, acoustic startle response, forced swimming, elevated plus-maze, passive avoidance were used for measuring emotional behavior and memory. The number of lymphocyte subpopulations CD3+, CD4+8-, CD4-8+, CD4+8+, CD19+, CD3+hi spleen and thymus, the ratio of cells in different phases of the cell cycle was determined by flow cytometry. C57BL/6J mice strictly increased anxiety in response to the KO compared to CBA mice. Moreover, KO-treated C57BL/6J mice impaired the passive avoidance learning. We found that KO evoked significant changes in the cellular composition of the thymus and decrease of thymocytes proliferation in C57BL/6J mice. In opposite KO-treated CBA mice showed change of splenic cellular structure with increased % CD19+ cells and the proliferation of splenocytes. Our study demonstrated genotype-dependent reactions of the nervous and immune systems in response chronic constant light.

[THE CLINICAL LABORATORY MARKERS OF ATHEROSCLEROSIS IN PATIENTS WITH ATHEROTHROMBOTIC STROKE].

The laboratory biomarkers can effect on choice of tactics of treatment in patients with atherosclerotic stenosis ofcarotids and high risk of stroke. However, nowadays there is no established laboratory criteria of significant atherosclerotic affection of internal carotid. The purpose of study was to investigate informativeness of biomarkers of atherosclerosis in clinical molecuIar panel of expertise system of determining risk of stroke in patients with significant stenosis of carotid. The study included patients with 50-90% atherosclerotic stenosis of internal carotid in acute period of atherothrombotic stroke or transitory ischemic attack (group 1), patients with stable 50-90% atherosclerotic stenosis of inner carotid having no vascular events during 30 days before engaging into study (group II) and group of healthy volunteers without atherosclerosis of inner carotid. The examination of patients included anamnesis collection, evaluation of neurological status, analysis of serum level of biomarkers of atherosclerosis (lipoprotein-associatedphospholipase A2 (LP-PL A2), serum protein A associated with pregnancy (PA PP-A), lipoprotein (a) (LP(a)), asymmetric dimethylarginine (ADMA), C-reactive protein detected by highly sensitive technique (hsCRP) and lipid spectrum of blood) using enzyme-linked immunosorbent assay, duplex ultrasound scanning of brachiocephalic arteries. The stroke risk factors of other etiology were chosen as exclusion criteria except atherothrombotic one. The Mann-Whitney and Kruskal-Wallis tests were applied to establish group differences. The Data Mining techniques were applied to establish patterns of analyzing sample. Out of 356 examined patients, 30 patients of group 1, 51 patients of group II and 16 healthy volunteers were included in the study. All patients were comparable by gender and age (50-80 years). The serum level of hsCRP and ADMA in the group of patients of acutest period of ischemic stroke was significantly higher than in groups of patients with stable stenosis and healthy volunteers (p < 0.05). The comparison between three groups established no statistically significant differences in serum concentration of PAPP-A, LP-PL A2 and LP(a). The ADMA, hsCRP and PAPP-A can be recommended for including into clinical molecular panel for personalized diagnostic of causes of stroke along with clinical anamnestic data. The serum level of ADMA and hsCRP significantly increases in acute period of atherothrombotic stroke. The analysis of levels of ADMA, hsCR and PPAPP-A interpreted with regard to clinical anamnestic data can be proposed for enhancing quality of diagnostic of causes of stroke.

Parameters of cellular and humoral immunity in experimental hyperthyroidism and its correction.

We demonstrate changes in cellular and humoral immunity in animals with experimental hyperthyroidism induced by chronic administration of potassium iodide (KI) solution. KI increased the weight and cellularity of the thymus and spleen and number of antibody-forming cells to sheep red blood cells and modified the relative content of T cell subpopulations. Phytosorption complex LimfoFit modifying cellular and humoral immunity affected only its individual parameters in hyperthyroid animals.

Effects of experimental desynchronosis on the organs of immune system in WAG and ISIAH rats.

We studied the influence of abnormal illumination regimen on cell composition of the central and peripheral organs of the immune system in ISIAH rats and control WAG rats. In ISIAH rats, 24-h illumination for 14 days led to more pronounced inhibition of cell proliferation and differentiation in the thymus and more pronounced decrease in splenocyte proliferation and T and B cell counts in the spleen in comparison with WAG rats; however, the level of antigen-presenting cells in the spleen of ISIAH increased. We concluded that ISIAH rats are more sensitive to abnormal illumination regimen than WAG rats. Twenty-four-hour illumination was associated with impairments of central differentiation of T cells and activation of systemic inflammation followed by impairment of differentiation regulation, which can aggravate metabolic dysfunctions in these animals.

Reactivity of the immune system and activities of PPARα and PPARγ in the liver under conditions of LPS-induced acute inflammatory response in hypertensive ISIAH rats.

The effects of LPS from E. coli on DNA-binding activities of PPARα and PPARγ in the liver and immune system parameters of were studied in hypertensive ISIAH rats and normotensive WAG rats. In ISIAH rats characterized by low basal level of PPARα, PPARγ, and HDL, the response of the peripheral immune system compartment to LPS was more pronounced and was not associated with decrease in DNA-binding activities of PPARα observed in WAG. Proinflammatory stimulus did not induce proliferative changes in the thymus of ISIAH rats, which can reflect impaired relationships between the central and peripheral organs of the immune system. The character of regulatory interactions between PPARα and immune cells can differ in various rat strains and depend on initial PPARα activity, HDL level, specific features of immune status, resistance to stress, and hormonal and metabolic background.

Cell cycle arrest in the thymus and spleen in male mice under conditions of chronic social defeat stress: effects of diazepam.

The effects of chronic social defeat stress on the percentage of cells in different phases of the cell cycle and in apoptosis in the thymus and spleen of male mice were studied by the method of flow cytofluorometry. In stressed males, thymus weight decreased, the percent of proliferating thymocytes was significantly lower, and the percentage of G0-G1 cells was higher than in intact males. Stress substantially reduced the percentage of splenocytes in the G0-G1 phase and apoptotic cells, but the percentage of S and G2-M cells and proliferation index significantly increased. Chronic administration of anxiolytic diazepam prevented the majority of the changes in the percentage of cells in different phases of the cell cycle, but apoptosis in the thymus increased under these conditions. Possible association between cell cycle disorders, impairment of cell immunity, and chronic anxiety developing under conditions of long-term social defeat stress is considered.

The influence of experimental desynchronosis on the morphofunctional characteristics of mouse immune system.

The aim of this study is to investigate the influence of experimental desynchronosis on the immune system and estimate benzo[a]pyrene (BP) influence on the immune reactivity of mice under the condition of desyncronosis (DS) in comparison with intact organisms. Mice (CBA X C57BL)F1 were kept under constant light during two weeks and natural alternation of darkness and light (control group). Animals of the two groups received benzo[a]pyrene solution in olive oil. The existence of diurnal variation of cell populations of lymphoid organs and blood, humoral immune response to SRBC were revealed. Changes of diurnal dynamics of these parameters and decrease of humoral immune response to SRBC have been observed in animals kept under constant light in comparison with animals kept under standard light conditions. It is likely to indicate the development of internal DS. BP influenced the cellular composition and immune response of intact and DS animals in a different way. So, the changes of light regime lead to development of the internal DS of immune system that influenced its functional capacity and sensitivity to damaging effect of chemical ecological factor.

Novel conjugate of moxifloxacin and carboxymethylated glucan with enhanced activity against Mycobacterium tuberculosis.

Mycobacterium tuberculosis is an intracellular pathogen that persists within macrophages of the human host. One approach to improving the treatment of tuberculosis (TB) is the targeted delivery of antibiotics to macrophages using ligands to macrophage receptors. The moxifloxacin-conjugated dansylated carboxymethylglucan (M-DCMG) conjugate was prepared by chemically linking dansylcadaverine (D) and moxifloxacin (M) to carboxymethylglucan (CMG), a known ligand of macrophage scavenger receptors. The targeted delivery to macrophages and the antituberculosis activity of the conjugate M-DCMG were studied in vitro and in vivo. Using fluorescence microscopy, fluorimetry, and the J774 macrophage cell line, M-DCMG was shown to accumulate in macrophages through scavenger receptors in a dose-dependent (1 to 50 microg/ml) manner. After intravenous administration of M-DCMG into C57BL/6 mice, the fluorescent conjugate was concentrated in the macrophages of the lungs and spleen. Analyses of the pharmacokinetics of the conjugate demonstrated that M-DCMG was more rapidly accumulated and more persistent in tissues than free moxifloxacin. Importantly, therapeutic studies of mycobacterial growth in C57BL/6 mice showed that the M-DCMG conjugate was significantly more potent than free moxifloxacin.

Circadian dynamics of cell composition of the thymus and lymph nodes in mice normally, under conditions of permanent illumination, and after melatonin injection.

The effect of melatonin on disturbed circadian variations in the lymphocyte subpopulation composition of the thymus and inguinal lymph nodes was studied in CBA mice exposed to constant illumination for 14 days. The desynchronizing effect of permanent illumination on the thymus consisted in disappearance of circadian variations in the total number of thymocytes, absolute count of thymocytes, absolute counts of CD8(+) and CD4(+) cells, and in inversion of changes in the absolute counts of CD4(+)8(+) cells from 15.00 to 20.00. In lymph nodes circadian variations in the percentage of CD4(+)+ lymphocytes disappeared, while absolute counts of CD4(+)8(+) and CD8(+) cells changed from 15.00 to 20.00. Melatonin restored circadian dynamics of some parameters mainly in the thymus.

Possible role of P-glycoprotein in cyclophosphamide resistance of transplanted mouse RLS lymphosarcoma.

The causes of different sensitivity of mouse LS lymphosarcoma and its resistant RLS variant to cyclophosphamide were studied. Division of LS and RLS cells stops in the G2/M phase 24 h after cyclophosphamide treatment, but this stop lasts for more than 48 h in LS cells and less than 24 h in RLS cells. DNA fragmentation, a marker of apoptosis, is observed only in LS cells starting from 24 h after cyclophosphamide treatment. LS and RLS strains do not differ by the expression of bcl-2, bcl-6, bax, bad, mdr1a, mdr1b genes and P-glycoprotein protein. The strains differ by transport activity of P-glycoprotein, tested by SYTO 16 substrate release from cells: activity of P-glycoprotein in RLS cells was 2-fold higher than in LS cells. Presumably, the resistance of RLS tumor to cyclophosphamide-induced apoptosis is a result of inhibition of the apoptotic cascade by P-glycoprotein which is functionally more active in these cells than in LS cells.

Functional activity of P-glycoprotein in lymphocytes of patients with lymphoproliferative diseases.

Functional activity of P-glycoprotein in lymphocytes of patients with lymphoproliferative diseases was studied using rhodamine 123. Functional activity of P-glycoprotein in patients receiving a course of chemotherapy was lower than in controls. P-glycoprotein activity was higher in patients receiving more aggressive therapy. Initially activity of P-glycoprotein was higher in patients who did not respond to chemotherapy in comparison with those whose clinical status improved after a course of chemotherapy.

[Effects of hypoxic stimulation in experimental animals and in children with bronchial asthma]. / Effekty gipoksicheskoi stimuliatsii u zhivotnykh, nabliudaemye v éksperimente, i u detei, bolnykh bronkhialnoi astmoi.

Inhalations using hypoxic mixture of 12-15% O2 were tested in 40 rats (daily for 20 min. during 1-4 months) and in 200 children with light and mid-serious forms of bronchial asthma (for 5-10 min. 2-4 times with 5 min. break, course-10 days). Stimulation effect in animals was observed for 1-2 months and then depression came. The immunological and allergological indices were observed to distinctly improve, whereas the indices of the pulmonary function remained practically unchanged. The clinical indices in most patients improved and the similarity of this method and effects of other nonmedicamental methods was noted.