Lactobacillus for Vaginal Microflora Correction.

INTRODUCTION: Despite the significant progress made in prevention, diagnosis, and treatment, there is still a high rate of vaginal dysbiosis in Kazakh women. The use of antibiotics in the treatment of vaginal dysbiosis contributes to the elimination of pathogens as well as microflora, which can lead to a decrease in local immunity and more favorable conditions for infection spread. The most physiologically safe and promising method for the restoration of vaginal biocenosis is the use of probiotics administered by a vaginal route. METHODS: We have allocated 64 of cultures of Lactobacillus from the vaginal epithelium of healthy women of reproductive age and women with diagnosed bacterial vaginosis (BV). Identification of cultures was performed by PCR analysis of 16S ribosomal RNA. Evaluation of biological significance was determined by the following criteria: high antagonistic activity against Candida albicans, Escherichia coli, Serratia marcescens, Proteus mirabilis, Klebsiella ozaenae, and Staphylococcus aureus; and production of hydrogen peroxide, resistance to antibiotics, adhesive activity. We studied the symbiotic relationship of selected biologically active of cultures to each other and received options for consortiums with properties of probiotics through co-cultivation. RESULTS: Results of genotyping showed that the isolated lactobacilli belong to the seven species: L. fermentum, L. salivarius, L. gasseri, L. crispatus, L. jensenii, L. plantarum, and L. delbrueskii. L. fermentum, L. salivarius, L. gasseri, and L. jensenii occur in women with suspected BV. The highest percentage of occurrence in the vagina of healthy women was L. fermentum (28%). Most strains of lactobacilli possess high inhibitory activity for all test-strains, except Candida albicans (37.5%). 56% of studied cultures revealed high adhesion to human erythrocytes. All lactobacillus strains were resistant to metronidazole, 80% to kanamycin, 57% to vancomycin, and sensitivity to roxithromycin, amoxiclav, ampicillin was diagnosed in all strains. 50% of cultures showed a moderate sensitivity to gentamicin and cefazolin. In a study of peroxide-producing activity, 80% of the cultures exhibited peroxide-producing activity. As a result of screening, the 7 most active strains of lactobacilli were selected for development of 10 variants of probiotic consortia. Also, there was increase of adhesive activity in the consortia compared to other components. These consortia can be used for the treatment of BV in addition to metronidazole. CONCLUSION: The probiotic consortia identified in this study had high antagonistic, adhesive properties, and resistance to metronidazole. These probiotics can potentially be used for the development of biological products for the treatment and prevention of bacterial vaginosis.

Study of the erythropoiesis activity of nano-encapsulated forms of erythropoietin.

INTRODUCTION: The recombinant human erythropoietin (rhEPO) is used in the treatment of anemia. In order to improve its pharmacokinetic properties, nanoparticles of biodegradable polymers of natural or synthetic origin were used. The aim of this study was to investigate the effect of new nano-encapsulated forms of recombinant human erythropoietin for oral use on the erythropoiesis in the cyclophosphamide immunosuppression model. MATERIAL AND METHODS: The CHOpE immortalized cells culture (a primary producer of rhEPO "Vector" in Russia) was used. The following biodegradable polymers were chosen: 0.05% and 0.005% carbopol, 0.05% and 0.005% kollidon, and 0.05% and 0.005% pectin. Immunosuppression was obtained by a single dose of i.p. injection of cyclophosphamide (250 mg/kg) in white mice (18-20 g). During the next 5 days, the nano-encapsulated erythropoietin (100 ED/mouse) was administered orally to each mouse. After 5 and 10 days, the cell count of the number of blood reticulocytes and the myelogram of bone marrow were performed. The control group of mice received injections of Eprex. RESULTS: On the 5th day of the experiment, the highest level of reticulocyte was observed in the samples of erythropoietin with kollidon (0.05%) and pectin (0.005%) nanoparticles. On the 10th day, the highest activity was observed in the samples of erythropoietin substance with pectin at 0.05% and 0.005% concentrations. The levels of reticulocytes in these groups reached 13.53% and 14.55%, respectively. The results of the myelogram during immunosuppression showed some activity of erythropoietin in conjunction with both concentrations of pectin when a two-fold increase in the number of erythroblasts was observed on the 5th day. High degrees of erythrokaryocytes in the state of mitosis were observed in the 0.05% pectin samples. Similar results were observed in equivalent groups of control animals on the 10th day of the experiment, which is compatible with the data on Eprex action. CONCLUSION: The erythropoietic activity of nano-encapsulated forms of erythropoietin was observed in the 0.05% and 0.005% pectin samples in the cyclophosphamide immunosuppression model setting.

Effect of Probiotic Consortium on the Local Inflammatory Process in Chronic Periodontitis.

INTRODUCTION: Inflammatory periodontal disease is one of the major concerns of researchers and clinicians, because it can lead to tooth loss and an increased risk of systemic pathologies, even at the age of 35. The purpose of this study was to determine the effects of gelatin-based probiotic consortium on the local and general factors of inflammation in rats with chronic periodontitis. METHODS: The study object was a complex of probiotic bacteria based in an odourless 6% gelatin plate with neutral flavour. A cellular biomass of the consortium consists of following lactobacilli: Lactobacillus casei subsp. pseudoplantarum, Lactobacillus caseisubsp.casei, L.fermentum, and L. helveticus. The viable cell number was 2.5 × 109 CFU/ml. The model of chronic periodontitis was reproduced in the white random-bred rats that weighed 160-220g, by keeping them on a low-protein diet. After three months, symptoms associated with medium and severe chronic periodontitis were observed in the rats. Application was carried out on the oral mucosa of rats 1 time per day for 14 days. The stickers lacking consortium of microorganisms were used as the placebo. The "Solcoseril" gel was chosen as a comparator. The hematologic, biochemical, and morphological characteristics were investigated. RESULTS: A complete clearance of periodontal pockets was observed during an objective examination of the experimental group rats on the 14th day of the experiment. Moreover, a gingival mucous turned pink, and there were no cyanosis tissues. The local changes were accompanied by improvement in hematological parameters, such as a reduction of blood eosinophilia and neutrophilia, and a recovery of the white blood cells number to the normal degree within the group that received the probiotic complex. A decrease of the acute plethora of microvasculature was observed morphologically as a result of the treatment. There were signs of basal layer activation of the stratified squamous epithelium with a merger of the acanthosis outgrowths and a formation of the fibrotic nodules. Biochemical investigations did not show significant changes in the indicators. CONCLUSIONS: In the settings of the chronic periodontitis model, the use of gelatin-based probiotic consortium consisting of Lactobacillus casei subsp. pseudoplantarum, Lactobacillus caseisubsp.casei, L.fermentum, L. helveticus. at 2.5 × 109 CFU/ml viable cell numbers lead to the reduction of the local inflammatory manifestations of the periodontitis in 14 days of treatment.

Health benefits of new symbiotic "NAR".

INTRODUCTION: The immune-modulatory effects of synbiotics and their ability to reduce free radical levels may be useful for functional food that is able to be active throughout whole period of colonization of the gastrointestinal tract.The aim of the present study was to investigate the immune-modulatory and antioxidant effects of the synbiotic product "NАR," a probiotic beverage. METHODS: The presence of IL-2, IL-4, IL-6, IL-8, IL-10, αTNF, γIFN, Ig A, Ig M, and Ig E was studied in vitro using a solid immunosorbent analysis. The total antioxidant activities of superoxide dismutase and glutathione reductase were determined by a spectrophotometry using the Sigma-Aldrich sets. RESULTS: Studies of the immune-modulatory properties of the synbiotic product NAR showed 1.7 fold increase of γINF levels (p<0.01) in blood after consumption of the synbiotic product "NAR" in comparison to control values, whereas the concentrations of IL-4 and Ig E decreased 2.0 times (treatment: 9.3; control: 18.7; p<0.01) and 1.3 times (p<0.1), respectively. The consumption of the synbiotic product "NAR" caused an increase in the proportion of γINF/IL 4 (treatment: 15.4; control: 4.4; p<0.01), which indicates a reduction in functional activity of Th2-type lymphocytes in comparison with the function of Th1 cells.Our study showed a high level of the total antioxidant activity of the synbiotic product (67.4 mmol/ml). The antioxidant activity of the intact cells of consortium (15.3 mM/ml), which was the basis for the preparation of the symbiotic product, is several times lower than the activity observed in the symbiotic samples.Expression of SOD is one of the mechanisms of antioxidant stress radicals inactivation by bacteria. The analysis identified a superoxide dismutase activity of synbiotic product (1.42 U/mg protein). A glutathione reductase activity of the synbiotic product was elevated (0.06 U/ml). CONCLUSION: The majority of the inflammatory mediators found in the blood after the consumption of symbiotic product NAR were inflammatory mediators that activate a cellular component of the resistance. Moreover, the symbiotic product has a high antioxidant activity.

Influence of Probiotic Consortium on TH1 and TH2 Immune Response.

INTRODUCTION: The main role of probiotics is to maintain homeostasis in the intestines and improve bowel protective function. The aim of the present study is to investigate immuno-modulatory effects of a probiotic consortium. METHODS: Observations were carried out in vitro. The presence of IL-2, IL-4, IL-6, IL-8, IL-10, TNF-α, IFN-γ, IgA, IgM, and IgE was studied using a solid-phase enzyme immunosorbent assay on the VECTOR-BEST sets (Russia). RESULTS: Immunomodulatory properties of the probiotic consortium were studied, which consisted of the following strains: Streptococcus thermophilus, Lactococcus lactis, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus acidophilus, Bifidobacterium longum, and Bifidobacterium bifidum. Elevated concentrations of INFγ in control samples decreased 3.9 times (p < 0.05) after a saturation of blood with the probiotic consortium. Significant reduction of cytokine levels after the probiotic effects of the consortium was observed in IL-10 by 2.1 times (p < 0.05) and IgA by 1.87 times (p < 0.0005). There was a significant increase in the levels of IL-4, IgE, IL-6, and IL-8 by 1.3 (p < 0.005), 1.1 (p < 0.5), 18.0 (p < 0.005), and 6 (p < 0.05) times, respectively, in comparison with the control samples. IL-4 and INFγ have different effects on the synthesis of IgE. Soluble low affinity receptors FcɛRII (CD23) in association with IL-4 facilitate a differentiation of the B-lymphocytes in IgE-synthesizing cells, while γ-INF inhibits this process. It is known that the intracellular expression of γ-INF and IL-4 is the most reliable marker for Th1 and Th2 immune responses, respectively. The conducted studies determined that the ratio of INF-γ/IL-4 was 0.9 (control 4.8, P < 0.005) after the saturation of the blood cells with probiotic consortium. NF-γ/IL4 ratio decreased by 5.3 times compared with a control value, which indicates a reduction in the functional activity of Th1 type lymphocytes in comparison with the function of Th2 cells. CONCLUSION: The application of the probiotic consortium results in the maintenance of homeostasis by the stimulation of immune function through the activation of humoral immunity. Moreover, the probiotic application changes the orientation of the immunological memory causing the cancellation of the recruitment of Th1 cells in the response.