[The role of microbiota and flavonoids in maintaining the balance of helper and regulatory T-lymphocytes associated with the intestinal immune barrier].

The gastrointestinal tract is a barrier, represented by dynamic and mutually regulating components (microbial, chemical, physical and immune) for the selective penetration of luminal contents into the internal environment. From the point of view of immunologists, even in a physiological condition, the epithelium of the intestinal wall is in a state of mild inflammation, which is explained by the constant invasion of antigens (food, microbial) and, in turn, the constant readiness of the immune system to respond. The purpose of this review was to analyze information about the formation of microbial and immunological barriers, immunological tolerance to microbiota and the possible role of flavonoids in this. Material and methods. The literature search was carried out using PubMed, ResearchGate, Elibrary databases mainly for the last 10 years, using the following keywords: flavonoid, gut microbiome/microbiota, Th17, Treg, RORγt, immunity, segmented filamentous bacteria. Results. During the immune response, a significant role in maintaining the intestinal barrier function is assigned to helper T lymphocytes type 17 (Th17). The intestinal microbiome is a key element in the formation of the immune barrier. Th17 differentiation in the intestine is fully triggered by commensals (apparently, the main role belongs to segmented filamentous bacteria) after weaning and the start of complementary feeding. Pro-inflammatory Th17 effectors in the gut are controlled by anti-inflammatory regulatory T-cells (Treg). In recent years, it has been established that despite the opposing functions of regulatory cells and effector Th17 cells, their differentiation is similar and is characterized by the expression of the common transcription factor RORγt. The main part of the peripheral regulatory lymphocytes of the intestine is a population that stably expresses not only FOXP3, but also RORγt. Flavonoids, which are plant secondary metabolites of the polyphenolic structure, are able to inhibit intracellular kinases and, as a result, influence the activation and implementation of effector functions of immunocompetent cells. Some flavonoids promote RORγt expression and appear to be able to reprogram the effector phenotype of Th17 cells, reducing their pathogenicity. Conclusion. Understanding the interactions between the microbiota, immune cells, and factors involved in their regulation, which are critical for the maintenance of tolerance, may facilitate progress in the prevention and therapeutic approaches to treat immunoinflammatory and autoimmune diseases.

Antiproliferative Effects of Modified Bioflavonoid in Ex Vivo Model.

We studied the effect of modified bioflavonoid and reference drug quercetin dihydrate on proliferation of mononuclears triggered by T- and B-cell mitogens. Lymphocytes were in vivo pretreated with the examined agents followed by their explantation and in vitro activation with T- and B-cell mitogens in cell culture. Intraperitoneal injection of modified bioflavonoid and quercetin dihydrate produced a dose-dependent inhibition of proliferation of the in vitroactivated splenocytes; modified bioflavonoid demonstrated higher antiproliferative activity.

Evaluation of the Effect of Modified Bioflavonoid and Quercetin Dihydrate on Cytokine Secretion by Mitogen-Activated Mononuclear Cells.

An experimental study revealed the effect on modified bioflavonoid on the inhibition of secretion of IFN-γ and IL-2 by ConA-stimulated mononuclear cells. These changes were accompanied by an increase in the secretion of IL-17 and IL-6. Our results suggest the differentiation of CD4(+) T helper cells into Th1 and Th17 subpopulations. The reference drug quercetin dihydrate induced an insignificant change in the level of IL-2 and IL-6 and small increase in IFN-γ content. The content of IL-17 was shown to decrease above the detection limit.

Species diversity and relative abundance of vaginal lactic acid bacteria from women in Uganda and Korea.

AIMS: Lactobacilli play an important role in maintaining vaginal health of women. The aim of this study was to compare the species richness and relative abundance of Lactobacillus and other lactic acid bacteria in women of two geographically distant countries, Uganda and Korea. METHODS AND RESULTS: Vaginal samples were obtained from two women populations in Uganda and Korea. The Lactobacillus Rogosa SL agar was used for initial isolation of lactic acid bacteria. After phenotypic analyses, the 16S rRNA gene was amplified by polymerase-chain reaction and analysed by the BLAST program and phylogenetic tree construction. A total of 338 (128 Korean and 210 Ugandan) vaginal lactic acid bacterial strains were isolated, including five genera: Lactobacillus, Leuconostoc, Pediococcus, Streptococcus and Weissella. While Lactobacillus crispatus was common in both populations, Lactobacillus fermentum was common only in Korean women, and Lactobacillus gasseri, Lactobacillus reuteri and Lactobacillus vaginalis only in Ugandan women. Among other lactic acid bacteria, Weissella was more common in Ugandan, and Pediococcus in Korean women. All Weissella strains produced hydrogen peroxide, and all Pediococcus strains inhibited Candida species. CONCLUSION: Although many lactic acid bacteria colonize women, their species distributions may be different in women of geographically separated communities. SIGNIFICANCE AND IMPACT OF THE STUDY: The knowledge of species richness and relative abundance of vaginal lactic acid bacteria, including Lactobacillus, Pediococcus and Weissella, may lead to the design of better probiotic products as bacterial replacement therapy.

Genetic diversity of vaginal lactobacilli from women in different countries based on 16S rRNA gene sequences.

AIMS: Lactobacilli are widely distributed in food and the environment, and some colonize the human body as commensal bacteria. The aim of this study was to determine the species of lactobacilli that colonize the vagina and compare them with those found in food and the environment. METHODS AND RESULTS: Thirty-five Lactobacillus strains from women from seven countries were isolated, and sequences from 16S rRNA genes were determined and compared with existing data in GenBank. A phylogenetic tree was achieved using the Neighbour-Joining method based on the analysis of 1465 nucleotides. The results showed that most vaginal isolates were L. crispatus, L. jensenii and L. gasseri. Some were L. vaginalis, L. fermentum, L. mucosae, L. paracasei and L. rhamnosus. Two isolates from a native American woman displayed distinct branches, indicating novel phylotypes. Few vaginal isolates matched food or environmental Lactobacillus species. CONCLUSIONS: Most women worldwide were colonized by three common Lactobacillus species: L. crispatus, L. jensenii and L. gasseri. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of vaginal Lactobacillus species richness and distribution in women worldwide may lead to the design of better probiotic products as bacterial replacement therapy.

Comparative study of vaginal Lactobacillus phages isolated from women in the United States and Turkey: prevalence, morphology, host range, and DNA homology.

Lactobacilli play an important role in maintaining vaginal health. However, during bacterial vaginosis lactobacilli decrease for unknown reasons. Our preliminary study showed that phages could infect vaginal lactobacilli. Therefore, the aim of this study was to analyze the distribution, virulence, and types of vaginal Lactobacillus phages isolated from women of two countries: the United States and Turkey. A total of 209 vaginal lactobacilli were isolated from reproductive-aged women in the United States (n = 107) and Turkey (n = 102). By analysis of 16S rRNA gene sequence and by comparison of protein profiles, most lactobacilli were identified as L. crispatus, L. gasseri, and L. jensenii. After mitomycin C induction, 28% of American lactobacilli and 36% of Turkish lactobacilli released phages. A total of 67 phages were isolated and further characterized by their host range, electron microscopy, and DNA homology. All 67 phages were infective against lactobacilli from both collections. The host ranges of most phages were broad, including multiple Lactobacillus species. Even though the phages were all temperate, they were able to cause lytic infection in various strains. The electron micrographs of these phages showed a hexagon-shaped head and a long tail with or without a contractile tail sheath. Based on their morphology, these phages belonged to Bradley's phage groups A and B, and could be further classified into four morphotypes. All four types were found among American phages, but only three were found among Turkish isolates. DNA hybridization with labeled probes of the four types of phages revealed that additional genetic types existed within each morphotype among these phages. The phage genomic sizes ranged between 34 and 55 kb. Many of the lysogenic Lactobacillus strains released phages spontaneously at a high frequency of 10(-3) to 10(-4) PFU/cell. In conclusion, lysogeny in vaginal lactobacilli is widely spread. Some lysogenic lactobacilli spontaneously release phages with a broad host range, which can be lytic against other vaginal lactobacilli regardless of their geographic origin.

In vitro inhibition of commercial douche products against vaginal microflora.

Recently, vaginal douching has been associated with many health risks in women. The aim of this study was to analyze the effect of commercial douche products against various vaginal microorganisms, including lactobacilli. Seven commercial douches were tested against eight Lactobacillus clinical isolates and three type strains from the American Type Culture Collection. BV-associated bacteria included six strains of five genera: Gardnerella, Mobiluncus, Mycoplasma, Peptostreptococcus, and Ureaplasma. Two isolates of group B Streptococcus, and three species of Candida were also tested. The minimal inhibition concentrations and minimal contact times for these products against vaginal microorganisms were determined in broth cultures. Four antiseptic-containing douche products showed a strong inhibitory effect against all vaginal microorganisms tested with a short contact time (less than 1 min). Three vinegar-containing douche products selectively inhibited vaginal pathogens associated with bacterial vaginosis, group B streptococcal vaginitis, and candidiasis, but not lactobacilli. The antimicrobial effects of the commercial douche products varied among different brands and microbial species tested.

Induction of vaginal Lactobacillus phages by the cigarette smoke chemical benzo[a]pyrene diol epoxide.

Because smoking increases a woman's risk of contracting bacterial vaginosis (BV), which is manifested by a reduction of vaginal lactobacilli and an overgrowth of anaerobic bacteria, chemicals contained in cigarette smoke were analyzed in vitro to determine their role in reducing lactobacilli. The result showed that trace amounts of benzo[a]pyrene diol epoxide (BPDE), which can be found in vaginal secretion of women who smoke, significantly increased phage induction in lactobacilli. This finding implies that smoking may reduce vaginal lactobacilli by promoting phage induction.

[The clinical profile of exogenous organic depressions in subjects who were victims of the accident at the Chernobyl Atomic Electric Power Station]. / Klinicheskii profil' ékzogenno-organicheskikh depressii u lits, postradavshikh v rezul'tate avarii na ChAES.

A total of 200 persons victims of the Chernobyl accident presenting with different exogenous organic depressions were examined. Radiation exposure did not exceed medium levels. The pattern of depressive syndrome of exogenous organic genesis was studied as was the incidence of its varieties. The article contains a description of different varieties of depression and sums up studies on premorbid and neurological status and additional research methods together with studies on suicidal intentions and disposition to alcoholization in the studied groups.

Phage infection in vaginal lactobacilli: an in vitro study.

OBJECTIVE: During bacterial vaginosis, an unexplained decrease of vaginal lactobacilli occurs. To identify whether these lactobacilli could be infected by phages, we isolated phages from vaginal lactobacilli and analyzed their potential virulence in attacking vaginal lactobacilli in vitro. METHODS: Vaginal samples were obtained from 39 reproductive-aged women. The selective Rogosa SL agar was used to isolate lactobacilli, from which phages induced by mitomycin C or released spontaneoulsy were analyzed by the agar spot method. RESULTS: Of 20 samples from women with vaginal infections, 12 did not have lactobacilli. From the remaining 8 infection samples and the 19 samples from healthy women, 37 Lactobacillus strains were isolated, from which 7 temperate phages were identified. Upon analysis, all 7 phages infected vaginal lactobacilli from the same and/or different women in vitro. Two phages, Phikc005 and Phikc007, had a broad host range, infecting 7 of 8 species tested. A control intestinal Lactobacillus phage also lysed several vaginal strains. One vaginal phage, Phikc039, was apparently lytic against vaginal lactobacilli from 7 other women. This phage was characterized as follows: plaque morphology, small and clear; burst size, 300 phages per cell; spontaneous induction rate, 1 per 10(6) cells; DNA, double-stranded and linear, 41 kb; and shape, a hexogonal head and a non-contractile tail. CONCLUSIONS: Bacteriophages were isolated from vaginal lactobacilli of some women and were shown in vitro to lyse vaginal Lactobacillus strains from the same and/or different women. It was suggested that vaginal lactobacilli might be suppressed by phages.

Analysis of lactobacillus products for phages and bacteriocins that inhibit vaginal lactobacilli.

OBJECTIVE: Bacterial vaginosis is associated with an unexplained loss of vaginal lactobacilli. Previously, we have identified certain vaginal lactobacilli-released phages that can inhibit in vitro other vaginal lactobacilli. However, there is no apparent route for phages to be transmitted among women. The purpose of this study was to identify whether certain Lactobacillus products commonly used by women release phages or bacteriocins that can inhibit vaginal lactobacilli. METHODS: From 26 Lactobacillus products (2 acidophilus milks, 20 yogurts, 3 Lactobacillus pills, and 1 vaginal douche mix), lactobacilli were isolated with Rogosa SL agar (Difco, Detroit, MI). From these lactobacilli, phages and bacteriocins were induced with mitomycin C and tested against a collection of vaginal Lactobacillus strains. RESULTS: From the 26 products, 43 Lactobacillus strains were isolated. Strains from 11 yogurts released phages, among which 7 inhibited vaginal lactobacilli. Eleven strains released bacteriocins that inhibited vaginal lactobacilli. While about one-half of the vaginal strains were lysed by bacteriocins, less than 20% were lysed by phages. CONCLUSIONS: Some vaginal lactobacilli were inhibited in vitro by phages or bacteriocins released from Lactobacillus products used by women, implying that vaginal lactobacilli may be reduced naturally due to phages or bacteriocins from the environment.

Analysis of Lactobacillus phages and bacteriocins in American dairy products and characterization of a phage isolated from yogurt.

Yogurt and acidophilus milk that contain Lactobacillus acidophilus could promote human health because L. acidophilus can inhibit enteric and food-borne microbial pathogens. To evaluate the stability of diary L. acidophilus cultures, we studied whether some diary lactobacilli could be inhibited by phages or bacteriocins released by other dairy lactobacilli. From 20 yogurts and two acidophilus milks purchased at local food markets, 38 Lactobacillus strains were isolated. Eight Lactobacillus type strains were used as controls. With mitomycin induction and agar spot assay, phages and bacteriocins were isolated from these strains and their activities were analyzed. Lactobacillus strains from 11 yogurts released phages, while the strains from most of the remaining products released bacteriocins. One phage, designated phi y8, was characterized. It was spontaneously released from its host strain L. acidophilus Y8, at a rate of about 10(4)/ml. This phage lysed nine other dairy Lactobacillus strains tested. It had a burst size of 100, an elongated prolate head of 39 by 130 nm, a long, flexible but noncontractile tail of 300 nm, and a 54.3-kb linear double-stranded DNA. DNA fingerprinting analysis indicated that L. acidophilus phages of nine yogurts in this study belonged to the same type as phi y8. Although they may be sensitive to bacteriocins, all lysogens resisted further phage attacks, whereas most nonlysogens were sensitive to both phages and bacteriocins. Therefore, Lacotbacillus cultures of some American yogurts and acidophilus milks may be unstable or unsafe because they can either be inhibited by phages or bacteriocins or release them to inhibit lactobacilli or other diary products.

[The pathogenesis of central nervous system functional disordersafter exposure to microwave radiation]. / Biokhimicheskie aspekty patogeneza narushenii funktsii tsentral'noi nervnoi sistemy pri mikrovolnovom obluchenii.

A rate of lipid peroxidation, content of biogenic amines and cyclic nucleotides were studied in various brain structures of 168 rats under conditions of microwave irradiation within 24 min at 46 mW/cm2. Total irradiation of animals was shown to result in distinct inhibition of monoaminergic activity of brain, especially of hypothalamus, in impairment of metabolic reactions, in exhaustion of the lipid antioxidative system of brain cortex and in stimulation of the contrainsular apparatus functions.

[Integration of the plasmid carrying the genes for tryptophan metabolism of Bacillus mesentericus into the chromosome of Bacillus subtilis]. / Integratsiia plazmidy, nesushchei geny triptofanovogo metabolizma Bacillus mesentericus, v khromosomu Bacillus subtilis.

Integration of several tryptophan operon genes from Bacillus mesentericus into the chromosome of Bac. subtilis was carried out. The genetic material exchange between these two species is usually impossible in the process of transformation, because of poor homology. The integration only took place after the genes of Bac. mesentericus were introduced in Bac. subtilis cells on a hybrid plasmid. Plasmid markers also integrated into the chromosome, together with heterologous DNA. The integration occurred in the region of Bac. subtilis tryptophan operon, which is likely to be due to retained homology between corresponding DNAs. The integration acts occurred during prolonged maintenance of chimeric plasmid in autonomously replicating form. We suggest to designate this phenomenon "retarded homologous recombination".

[Genetics and phenogenetics of the hormonal characteristics of animals. VII. Correlation between brain serotonin and the hypothalamo-pituitary-adrenal system in domesticated and undomesticated silver foxes during emotional stress]. / GEnetika i fenogenetika gormonal'nykh kharakteristik zhivotnykh. VII. Korreliativnaia vzaimosviaz' mezhdu serotoninom mozga i gipotalamo-gipofizarno-nadpochechnikovoi sistemoi usloviiakh émotsional'nogo stressa u domestitsiruemykh i nedomestitsiruemykh serebristo-chernykh lisits.

Two groups of silver foxes, selected according their behaviour with respect to man and non-selected ones, were exposed to restriction-induced stress. It was found that changes in the level of brain serotonin and its main metabolite, 5-hydroxyindolacetic acid, and elevation of plasma corticosteroids concentration in domesticated (tame) silver foxes were much less pronounced than in non-domesticated (non-tame) animals. Positive relationship between type of behaviour, brain serotonin metabolism and pituitary-adrenal axis response to stress was observed. It is suggested that such correlative pattern as changed pituitary-adrenal axis responses in domesticated animals may be due to changes in metabolism of controlling this axis brain transmitter serotonin.

[Genetics and phenogenetics of the hormonal characteristics of animals. VI. Functional activity of certain chemoreceptors connected with the pituitary-adrenal system of silver foxes selected for their behavior]. / Genetika i fenogenetika gormonal'nykh kharakteristik zhivotnykh. Soobshchenie VI. Funktsional'naia aktivnost' nekotorykh khemoretseptorov, sviazannykh s gipofizarno-nadpochechnikovoi sistemoi u serebristo-chernykh lisits, selektiruemykh po povedeniiu.

Seasonal differences in the reaction of the pituitary-adrenal system in domesticated and non-domesticated silver foxes of both sexes to substances activating alpha-, beta-adrenoreceptors, and serotonin receptors were studied. It was shown that the reactivity of the pituitary-adrenal system in silver foxes of either type of behaviour is due, at least partially, to seasonal differences in the state of adrenergic and serotoninergic mechanisms. At the same time, in silver foxes selected for behaviour to man the reaction of the pituitary-adrenal system to the injection of substances activating adrenergic and serotoninergic receptors differs, during the year, from the reaction to these compounds in non-selected animals. The conclusion was made, that in the process of domestication changes take place in the state of serotonin- and noradrenaline mechanisms connected with the regulation of the hypothalamo-pituitary-adrenal complex.

[Changes in coagulographic indices of patients with cerebral atherosclerosis complicated by mental disorders during the treatment process]. / Izmeneniia nekotorykh pokazatelei koagulogrammy u bol'nykh tserebral'nym aterosklerozom s psikhicheskimi narusheniiami v protsesse lecheniia.

Examination and treatment of 40 patients with atherosclerotic dementia demonstrated a significant increase in some indices of the hemocoagulation system. These changes were expressed in an increase of blood tolerance to heparin, shortening of the recalcification time, increase in the fibrinogen content, presence of fibrinogen B. The group studied was characterized by the presence of slightly marked fibrinolysis compensatory reactions. Treatment by novocain combined with aspirin led to appreciable changes in these indices towards normalization, permitting to consider this form of therapy as pathogenetic.

[Changes in the blood clotting system of patients with atherosclerotic and senile dementias]. / Izmeneniia svertyvaiushchei sistemy krovi u bol'nykh ateroskleroticheskimi i starcheskimi dementsiiami.

The author studied changes of the coagulative blood system in patients with atherosclerotic and senile dementia (82 cases). In cases with simple forms of senile dementia a tendency towards hypercoagulability was established (an increased formation of thrombin, the amount of fibrinogen content). However, this group demonstrated a defensive activation of the anticoagulative system, sufficient to maintain an equilibrium in the whole system of hemocoagulation. An exhaustion of compensation may lead to stasis and microthrombosis in the smallest vessels and capillaries, which indicates to the pathogenetical significance of changes in the coagulative blood properties.