[Epithelial protective therapy in comorbid diseases. Practical Guidelines for Physicians].

This document was produced with the support of the National Medical Association for the Study of Comorbidities (NASС). In 2021 the first multidisciplinary National Consensus on the pathophysiological and clinical aspects of Increased Epithelial Permeability Syndrome was published. The proposed guidelines are developed on the basis of this Consensus, by the same team of experts. Twenty-eight Practical Guidelines for Physicians statements were adopted by the Expert Council using the "delphic" method. Such main groups of epithelial protective drugs as proton pump inhibitors, bismuth drugs and probiotics are discussed in these Guidelines from the positions of evidence-based medicine. The clinical and pharmacological characteristics of such a universal epithelial protector as rebamipide, acting at the preepithelial, epithelial and subepithelial levels, throughout gastrointestinal tract, are presented in detail.

Anti-Influenza Activity of Enterocin B In vitro and Protective Effect of Bacteriocinogenic Enterococcal Probiotic Strain on Influenza Infection in Mouse Model.

In spite of scientific evidence demonstrating the antiviral activity of lactic-acids bacteria, little is known about the mechanism of their action. Previously, several bacteriocins isolated from lactic acid bacteria (LAB) and some other microorganisms were reported as having antiviral activity in vitro. In the present study, chemically synthetized enterocin B (EntB) and the strain E. faecium L3, known as the producer of this peptide, were tested for activity against influenza viruses. The inhibition of cytopathic effect of А/Perth/16/2009(H3N2) and A/South Africa/3626/2013(H1N1) pdm influenza viruses in MDCK cells by chemically synthetized EntB was revealed. The EntB demonstrated antiviral activity at a concentration of 2.5-5 µg/ml depending on the dose of viruses. This peptide exhibited low toxicity in MDCK cells, causing partial damage of the monolayer of the cells only at a concentration above 10 µg/ml. It was also shown, that strain E. faecium L3-protected mice from lethal A/South Africa/3626/2013(H1N1) pdm infection. We speculate that this protective effect of enterococci may be associated with the specific action of enterocin B, which possesses antiviral activity in vitro.

[The intestinal microbiota composition in patients with multiple sclerosis receiving different disease-modifying therapies DMT]. / Osobennosti sostava mikrobioty kishechnika u patsientov s rasseiannym sklerozom, poluchaiushchikh preparaty, izmeniaiushchie techenie rasseiannogo skleroza.

AIM: To describe characteristics of the intestinal microbiota in patients with multiple sclerosis (MS) treated with glatiramer acetate (GA) or fingolimode (FG) for understanding causal relationships between gut microbiota and autoimmune processes in MS patients. MATERIAL AND METHODS: The study included 34 patients treated with GA (n=17) or FG (n=17). GA was used in a dose of 20 mg/kg subcutaneously once a day, FG in a dose of 0.5 mg daily. All patients were examined during remission. To assess the composition of gut microbiota, bacteriological and real-time PCR techniques were used. DNA was extracted from feces using DNA-EXPRESS kit. RESULTS AND CONCLUSION: There was a decrease in numbers of Escherichia coli with normal enzymatic activity, which was replaced by atypical forms of E. coli, Enterobacter spp. and fungi of the genus Candida, and, during treatment with GA, by atypical forms of E. coli, Proteus spp., Parvimonas micra. These differences indicate the effect of the therapy on the intestinal microbiota composition.

Modeling of 3D Structure of Chimeric Constructs Based on Hemagglutinin of Influenza Virus and Immunogenic Epitopes of Streptococcus Agalactiae.

A project of an experimental recombinant vector vaccine for prevention of diseases caused by pathogenic streptococci based on ScaAB lipoprotein of Streptococcus agalactiae and a coldadapted strain of live influenza vaccine as a vector was developed. The sequence of ScaAB lipoprotein was analyzed and fragments forming immunodominant epitopes were determined. Chimeric molecules of influenza virus hemagglutinin H7 carrying insertions of bacterial origin were constructed. Based on the results of simulation, the most promising variants were selected; they represented fragments of lipoprotein ScaAB lacking N-terminal domain bound to hemagglutinin via a flexible linker. These insertions should minimally modulate the properties of the influenza strain, while retaining potential immunogenicity to a wide group of pathogenic streptococci.

Inactivation of M111 Protein Gene Modifies Streptococcus Pyogenes Interactions with Mouse Macrophages In Vitro.

Immunomodulatory properties of S. pyogenes protein M111 were studied on the model of Gurov strain and its isogenic mutant not expressing M protein. Mouse resident peritoneal macrophages were incubated with bacteria and generation of nitroxide and superoxide anions and production of IL-6, IL-10, and IL-17 were evaluated. Protein M111 modified macrophage response: it exhibited antiphagocytic activity, prevented ROS formation, and stimulated the production of anti-inflammatory cytokine IL-10. The results suggested that this protein could serve in the bacteria as a factor suppressing the host defense forces and promoting the realization of the strategy beneficial for pathogens - escape from the host immune defense.

[The characteristics of microbiota of periodontal recesses in smoking patients with chronic generalized periodontitis].

The clinical examination of 36 tobacco smokers with chronic generalized periodontitis of light, average and severe degree was carried out. The examination established poor hygienic condition of oral cavity, less expressed inflammatory reaction of tissues of periodont and predominance of occurrences of destruction of alveolar portion of bone as compared with the group of 59 non-smoking patients with chronic generalized periodontitis of light, average and severe degree. The study demonstrated higher rate of detection of T. forsythia in smokers as compared with non-smoking patients at all stages of of development of chronic generalized periodontitis. Under light stage of chronic generalized periodontitis increasing of rate of detection of T. forsythia more than twice was registered. P.gigngivalis and P.intermedia were detected in smoking patients with light stage of chronic generalized periodontitis either in the same values or more rarely as compared with non-smokers. In the group of smokers with average stage of chronic generalized periodontitis increasing of rate of occurrence of association of T. forsythia-P. gigngivalis-P. intermedia occurred more than five times in comparison with non-smokers. The obtained results indicate on relationship between alterations of microbiota and aggressive development of chronic generalized periodontitis in smoking patients and on development in periodontal recesses of smokers of favorable conditions for growth of T. forsythia. The presence of T. forsythia is a significant factor of development of destructive processes in tissues of periodont.

Enterococcus faecium strain L-3 and glatiramer acetate ameliorate experimental allergic encephalomyelitis in rats by affecting different populations of immune cells.

The effect of probiotic Enterococcus faecium strain L-3 was studied in rats with experimental allergic encephalomyelitis (EAE). Glatiramer acetate (GA) was used as control drug. E. faecium strain L-3 and GA both were able to reduce the severity of EAE in a similar fashion. Both approaches increased the proportion of EAE resistant rats and rats with mild disease, prolonged the inductive phase of EAE and reduced the disease duration. Study of the phenotypes of immune cells in blood revealed the differences in immunoregulatory pathways that mediate the protective action of probiotic or GA treatment of EAE. The presence of pronounced protective and immunomodulating effects of the probiotic E. faecium strain L-3 opens an opportunity of its application for the treatment of multiple sclerosis.

[EFFECT OF PROBIOTIC ENTEROCOCCI AND GLATIRAMER ACETATE ON THE SEVERITY OF EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS IN RATS ].

Currently intestinal microbiota is considered as a potential target for influence in various pathologies which have inflammation, autoimmunity or neurodegeneration in the genesis. Multiple sclerosis (MS) combines all these processes in the pathogenesis. Furthermore, the balance of the components of intestinal microbiota is disrupted during MS and followed by disbiosis. Different probiotics - bacteria with proven beneficial properties are widely used to correct dysbisis. In this paper, was investigated the ability of probiotic strain Enterococcus faecium L-3 to reduce disease severity in multiple sclerosis model - experimental allergic encephalomyelitis (EAE). E. faecium L-3 were used alone or in combination with glatiramer acetate (GA). It is shown that administration of E. faecium L-3 reduces the severity of EAE in rats almost as same as that of GA. However, when the probiotic enterococci administered together with GA the protective effect does not observed. It is assumed that these preparations stimulates different ways of the immune system, because their action stimulate different immune cells populations. The study demonstrates the ability of E. faecium L-3 to influence on the immune system in MS, directly and indirectly (through the correction of dysbiosis). This fact allows us to consider E. faecium L-3 as a potential tool for immunomodulation in autoimmune, inflammatory and neurodegenerative diseases.

[INFLUENCE OF PROBIOTIC STRAIN E. FAECIUM L3 ON MYOCARDIAL TOLERANCE TO ISCHEMIA-REPERFUSION INJURY IN THE MODEL OF ANTIBIOTIC-INDUCED INTESTINAL DYSBIOSIS].

Functional parameters and degree of myocardial damage by ischemia-reperfusion model of isolated heart were investigated during acute experiments on rats with an antibiotic-induced intestinal dysbiosis after administration of the probiotic strains of Enterococcus faecium L3. Administration of antimicrobial drugs has resulted in an increased heart rate and in an increase in coronary flow of isolated heart. Administration of enterococci after the global ischemia has caused rats to have a decrease in diastolic pressure, increased systolic and pulse pressure, and an increased intensity of coronary flow. The article presents data on changes in the composition of the intestinal microbiota and the immune system that was previously obtained in studies based on this experimental model.

[PROBIOTICS, NUTRITIONAL STATUS AND RESISTANCE TO RESPIRATORY INFECTIONS IN INFANTS].

Aim of this study was to investigate the efficacy of prophylactic use of a liquid probiotic form based on Enterococcus faecium L3 in children first year of life to increase resistance to acute respiratory infections (ARI) in the winter-spring period and improve nutritional status. In a specialized (psychoneurologic) orphanage of St. Petersburg within three months of winter-spring period in 2014 observed 29 children in the first year of life. Observed children were randomly divided into two groups. The main group (n = 14) within three months received daily per os liquid probiotic form based on E.faecium L3 in a daily dose of 1.5 x 10(9) CFU. A control group of children (n = 15) was comparable with the main group by gender and age. The children observed groups studied the incidence of ARI; number of vaccinated children; the number of children who received two or more vaccinations at the same time; the number of children with acute infectious diseases within 1 month after vaccination; dynamics of Chulitskaya index, body mass index, body weight and length. The data obtained are subjected to statistical analysis, the results were considered significant at p < 0.05. The results showed that the use of probiotic forms E.faecium L3 in infants helped to reduce the average number of ARI cases per child (0.29 ± 0.13 vs. 0.73 ± 0.12 in the control group; p < 0.05) only in the first month of the observation that combined with a significantly higher BMI values increase this month (0.54 ± 0.25 vs. 0.07 ± 0.22 kg/m2 in the control group; p < 0.05). Use the liquid probiotic form E.faecium L3 in infants had a positive impact on overall health, which is reflected in the implementation of routine vaccination activities--in the main group, the number of children who received two or more vaccinations at the same time was 18.1% more.

[CHANGES IN THE QUALITATIVE AND QUANTITATIVE COMPOSITION OF GUT MICROBIOTA IN RATS DURING EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS].

In this study, on the model of multiple sclerosis - experimental allergic encephalomyelitis (EAE), the dynamics of changes in the qualitative and quantitative composition of the intestinal microbiota in rats with symptoms of the disease and asymptomatic course were compared. It was found that the composition of the intestinal microbiota in rats with the clinical symptoms of EAE is shifted towards gram-negative opportunistic microorganisms of the genus Citrobacter, Prote- us, Klebsiella and enteropathogenic Escherichia coli. It has been shown that rats without clinical signs of EAE have higher levels of Faecalibacteriumprausnitzii. The significance of the complex changes in the composition of the intestinal microbiota, indicating long-lasting dysbiosis in rats during the development of EAE is discussing.

[Problem of the choice of the probiotic dose in the physician practice].

Currently, there is ample evidence of the usefulness of probiotics for human health, but, despite this, progress in the scientific and medical recognition of the legal force (legalization) of probiotic agents remains extremely slow. Probiotic preparations are prescribed with the preventive and medical purpose, must be not only safe, but always effective, because these properties affect the economic component of medical activity. Clinical efficacy of probiotics is determined not only by the specifics of strains, but also the adequacy of daily and course dose that should not be underestimated. Recent studies prove the identity of the impacts on human health probiotic agents and functional food products containing probiotic strains in optimal concentrations. Definition of preventive and curative doses of probiotics in children in different age periods remains a challenge for pediatricians.

[Value of probiotic in reducing frequency of infectious complications in premature infants].

Fifty five premature infants with very low body weight were observed as inpatients to assess effectiveness of the nursing. The standard program of the care included: adequate primary resuscitation, respiratory support (use of surfactant), thermal mode, early provision of nutrients with full or partial parenteral nutrition, antimicrobial therapy and if possible early enteral nutrition. The patients of the main group (n = 29) along with the standard therapy (the volume of the enteral nutrition of 5 ml or more) were treated orally with liquid probiotic based on E. faecium L3 in a dose of 0.5 ml (5 x 10(8) CFU) 3 times a day for 14 days. The control group (n = 26) was under the standard therapy. The effectiveness of the infants management was estimated by the frequency of disruption of the nutrition, the frequency of infectious complications, the changes in the hematologic indices, the dynamics of the composition of the intestinal microflora. The infectious complications were evident from perinatal viral infection, intraamniotic infection, necrotizing enterocolitis. In the main group patients there was recorded a significant decrease of the frequency of infectious complications: 20.7% against 53.9% in the control group patients. The use of probiotic strain E. faecium L3 in the preterm infants promoted conservation of the immunomodulatory function of the intestinal indigenic microflora and restricted the growth of the nosocomial flora.

[Intranasal co-administration of recombinant streptococcus group B polypeptides and influenza deltaNS1 vaccine].

In the present work, the immunoadjuvant properties of the influenza deltaNS1 vaccine virus after intranasal administration in combination with recombinant GBS polypeptides was tested in mice. According to our data, co-administration of recombinant GBS polypeptides and influenza deltaNS1 vaccine resulted in the increase in the immunogenicity and protective efficacy of bacterial proteins. Combined vaccination with the GBS polypeptides and influenza deltaNS1 vaccine has a potential to be used not only for prophylaxis infections caused by SGB, but also for prevention of the bacterial complications of influenza.

[Probiotic strains of enterococci as a means of therapy and prophylaxis of intestinal diseases in children (review of literature)].

Probiotic enterococci a widely used by pediatricians and infection diseases doctors in Russia as means for the treatment of dysbiosis, irritated bowel syndrome and in the treatment and prevention of different functional and chronic intestinal diseases. Strains E. faecium M74 and E. faecium SF-68 are included in several probiotic drugs and have been proved as effective and safe. Several studies suggest that the usage of Russian probiotic strain E. faecium L3 in pediatrics is promising. The review is devoted to the analysis of clinical studies in pediatric practice employing probiotic enterococci.

[Autochthonous probiotics in prevention of infectious and inflammatory diseases of a human in the altered habitats].

Dysbiotic shifts in intestinal and pharyngeal microflora were studied in 22 normal volunteers in 9-, 14-, 105- and 520-d chamber experiments simulating some of the spaceflight factors. Two preparations were administered to prevent pharyngeal and intestinal dysbiosis: oral dry probiotic based on indigenous intestinal Enterococci and topical collagen-immobilized Lactobacterin based on indigenous intestinal Lactobacilli. Topical autopmrobiotic lactobacterin reduced the growth of opportunistic pathogens in the throat during the experiments. Oral autoprobiotic based on Enterococci reduced the content of intestinal opportunistic pathogens, supporting the high level of protecting microflora. Most of autostrains are free from pathogenicity factors, nevertheless, implementation of genetic testing of indigenous strains are reasonable.

[Distribution and genetic organization of the pathogenicity island XII among the clinical strains of GBS].

Streptococcus agalactiae or group B streptococci (GBS) is the major cause of various diseases of the newborns and the adults. The genome of GBS contains from 14 to 15 mobile genetic elements (pathogenicity islands, PAI). It is well-known that many GBS virulence determinants are localized not in the core genome, but on the pathogenicity islands. The goal of this work was to determine the distribution and genetic organization of PAI XII containing virulence genes sspB1, scpB, lmb among the clinical strains of GBS. 74 clinical strains of GBS were analyzed using PCR with primers corresponding to the genes of the virulence factors located on PAI XII. The pathogenicity island XII was determined only in 22% of the clinical strains. The genetic organization of the island was different between strains. There was no correlation between the presence of PAI and the serotype of GBS.

[Dynamics of contamination and persistence of Clostridium difficile in intestinal microbiota in newborn infants during antibiotic therapy and use of probiotic strain enterococcus faecium L3].

Ninety four infants were observed as inpatients. Thirty nine of them were mature neonates and 55 were premature infants with a very low body weight. The majority of the patients were treated with antibiotics. The mature infants were treated with penicillins, aminoglycosides, cephalosporins and the premature neonates were treated in addition with carbapenems, fluoroquinolones, glycopeptides. The mature infants were randomized into 2 groups: the control group (n=18) received the standard therapy and the main group (n=21) in addition received 1 ml of liquid probiotic Enterococcus faecium L3 (with a titer of 5x10(8) CFU/ml) 2 times a day for 10 days. The premature newborn infants were also randomized into 2 groups. The control group (n=26) received the standard therapy. The main group (n=29) additionally received 1 ml of liquid probiotic E.faecium L3 2 times a day for 10 days. The effectiveness of the therapy in the mature neonates was evaluated by the frequency of dyspeptic disorders and in the premature infants by the frequency of infectious complications and the episodes of food intolerance. The intestinal microbiota of the infants was investigated by the real-time PCR and bacteriological analyses of the feces: in the mature infants on admission to the hospital and 10 days after the treatment (periods 1-2), in the premature infants on admission to the hospital and then twice with an interval of 14 days (periods 1-2-3). It was shown that the use of the probiotic strain E.faecium L3 during the antibiotic therapy in the premature infants promoted significant reduction in the frequency of infectious complications. In the mature neonates the probiotic therapy reduced the risk of dyspeptic disorder. The studies showed reduction in persistence of Clostridium difficile in the intestinal microbiota of the newborn infants receiving the antibiotic therapy in combination with probiotic E.faecium L3, that was accompanied by preserving and growth of bifidobacteria and lactobacilli and reduction of the number of opportunistic microorganisms.

[Inhibition of herpes simplex virus type 1 reproduction by probiotic bacteria in vitro].

Light and immunofluorescence microscopies were used to study the cytopathic effect of herpes simplex virus type 1 (HSV-1) grown on the Vero cell cultures in the absence or presence of supernatants of Enterococcus faecium L3, Lactobacillus plantarum 8A-P3, and Escherichia coil M17. The effect of the probiotic strains was evaluated estimating the proportion of changed cells and the infective dose of the virus. The supernatants of the cultures of Lactobacillus sp. and Enterococcus sp., unlike those of E. coil, have antiviral activity. Inhibited viral replication was more evident when the supernatants were added until the cultured HSV-1 cells were infective. An enterococcal supernatant and its obtained peptide extract showed the maximum antiviral activity. This strain may be associated with the production of bacteriocins and bacteriocin-like substances.