One of the leading causes of the occurrence of chronic bacterial prostatitis (CBP) in men is infection, microecological disorders of the urogenital tract and cytokine-mediated mechanisms of ...inflammation of the prostate gland, which actualizes a comprehensive study of the clinical and bacteriological features of CBP from the perspective of a symbiotic approach in the framework of a new scientific field - "infectious symbiology".
to study the characteristics of spermogram, microbiota, and the cytokine profile in men with chronic bacterial prostatitis (CBP) and CBP complicated by infertility.
A comprehensive study of patients with CBP and CBP complicated by infertility, in comparison with conditionally healthy individuals, was conducted. Species identification of microorganisms was carried out according to biochemical characteristics and the genetic method (sequencing of strains). The biological properties of the microbiota were evaluated: growth properties, biofilm formation, antipeptide activity against the cytokines IL-10, RAIL-1, TNF-, INF- and IL-17 (8 parameters). Immunological parameters of sperm plasma included 13 parameters: the content of cytokines TNF-, INF-, Rail, interleukins (IL) -1, 2, 4, 6, 8, 10, 17, immunoglobulin (Ig) A, lactoferrin and lysozyme. To evaluate sperm plasma, the following quantities were determined: ejaculate volume, pH, sperm plasma liquefaction, total sperm count, sperm count per 1 ml, motility, number of progressively motile, non-progressive motile and motionless spermatozoa, number of round cells, white blood cells, spermatogenesis cells, erythrocytes, erythrocytes, cells, sperm agglutination and aggregation (16 parameters in total). The results are statistically processed.
Data were obtained on changes in biofilm formation, antipeptide activity of microbiota (especially pronounced in corynebacteria), sperm plasma cytokine profile (increased TNF , IL-2, 6, 17), as well as IgA and lactoferrin, which can be used to build a prognostic model of reproductive pathology tract of men and their fertile activity.
The study of the antipeptide activity of microbiota in combination with the cytokine profile of ejaculate allows us to recommend them as a "biotarget" for diagnostic, preventive and therapeutic measures for chronic prostatitis in men, which contribute to solving the medical and social problem of preventing male infertility and contributes to the development of health-saving technologies with incorporating elements of personalized medicine.
The culture liquid of lactic-acid bacteria (LABs) and bifidobacteria contained secondary metabolites fulfilling signal functions, such as guaiacol and γ-butyrolactone. In addition, 4-hexylresorcinol ...was detected in the cell extract of
Bifidobacterium bifidum
791. No homoserine lactones were produced by the tested microorganisms. The presence of metabolites with signal functions in conjunction with the available data on the synthesis of neuroactive amines, amino acids, and prebiotics by LABs and bifidobacteria indicates that these bacteria can strongly influence the functioning of the whole human symbiotic microbiota as well as the human host’s health state.
Aim. To determine the role of the acetate in the persistence of indigenous bifidobacteria in the colon biotope through the lysozyme resistance in model conditions of the acetylation–deacetylation of ...peptidoglycan.Materials and methods. The study was performed on 16 strains of the two indigenous bifidobacteria speсies: Bifidobacterium bifidum и Bifidobacterium longum subsp. longum. Bifidobacteria was cultivated in the 0.6% O2 and 9% CO2 atmosphere at the temperature 37ºС in CO2 incubator for 48 hours. The production of the acetate by the bifidobacteria was determined by gas chromatography. The effect of the acetate on the lysozyme resistance of non-indigenous gram-positive bacteria was determined on the Listeria monocytogenes ICIS-280 model strain by the cultivation in LB-Lennox broth with ammonium acetate added in the concentration range matching the concentrations produced by the studied bifidobacteria, in lysozyme serial dilutions at final concentrations 5 μg/ml to 40 μg/ml within 24 hours.Results. It was found that the acetate release of Bifidobacterium longum subsp. longum was on average two times higher that of Bifidobacterium bifidum (27.0 and 14.7 mmol/liter, respectively) and was quite consistent with the concentrations of acetic acid determined in the intestinal contents (up to 50 mmol/liter). Cultivation of bifidobacteria in a medium with lysozyme, ammonium acetate and their combination did not have a significant impact on their growth parameters at the maximum used concentrations of these substances. In the test strain, the addition of ammonium acetate in the range created by bifidobacteria caused a decrease in the minimum inhibitory concentration of lysozyme by more than two times — from 40 μg/ml to less than 20 μg/ml. In the control medium without lysozyme, no inhibition of the growth of the indicator culture was observed up to the maximum concentrations of ammonium acetate.Conclusion. The mechanism of persistence (survival) of indigenous bifidobacteria in the human intestinal biotope has been identified, which is associated with the production of acetic acid at a level that selectively suppresses lysozyme resistance of non-indigenous gram-positive microbiota viareversible deacetylation of peptidoglycan. This allows indigenous bifidobacteria to maintain a stable dominant position in the biotope.
Analysis of the study is to assess the diagnostic significance of cytokines in the sperm plasma of men of reproductive age (20 - 45 years) of two groups: of patients with chronic bacterial ...prostatitis, not complicated by infertility and with loss of fertility. The study of sperm plasma - the WHO standard. Determination of the level of cytokines in seminal plasma - by enzyme immunoassay («Cytokine», Russia). Two methods of mathematical statistics were used: discriminant analysis and classification trees (decision trees).The similarity of interpretations of discriminant analysis and decision tree was noted, where the main role in both cases belongs to the cytokine IL-4. The level of sperm IL-4 in combination with therapeutic monitoring can be used for the medical management of patients with chronic prostatitis in order to prevent the development of infertility and to develop methods for screening diagnostics of fertility disorders in men.
The most important role in homeostasis of intestinal immune belongs to the immunoregulatory properties of the microbiota which activates intracellular signaling systems, cytokine expression, ...production of protective factors and limits inflammatory reactions in the intestine by interacting with the pattern recognition receptors. The outcome of interactions between the microbiota and host cells (development of an inflammatory process or maintenance of intestinal homeostasis) depends on many factors, including a potential ability of intestinal commensals to influence the cytokine network in human body. Due to disturbances of quantitative and qualitative microbiota profile (dysbiosis), the cytokine balance may be changed by the influence of intestinal microsymbionts and their metabolites on immune and epithelial cells of intestines, thus contributing to the development of various human disorders. The aim of this study was to evaluate the immunoregulatory properties of eubiotic and dysbiotic human intestinal microsymbionts by assessing the effects of their cell-free supernatants on cytokine production in the
in vitro
system. The study was conducted on 49 eubiotic and 77 dysbiotic strains of microorganisms isolated from conditionally healthy patients examined for colon dysbiosis. To assess immunoregulatory properties of intestinal microsymbionts, we studied the effects of cell-free supernatants from bacterial and fungal cultures up on production of proinflammatory (IFNγ, TNFα, IL-17, IL-8, IL-6) and anti-inflammatory (IL-10, IL-1ra) cytokines secreted by mononuclear cells isolated from peripheral blood of healthy persons. The intestinal microbiota was determined by bacteriological methods. Identification of isolated microbial cultures was performed using MALDI TOF MS Microflex LT series (Bruker Daltonics, Germany). The level of cytokines was determined by enzyme immunoassay using commercial test systems (“Cytokine”, Russia). Statistical evaluation included discriminant analysis, classification decision tree and resultant mapping method. The multivariate statistical analysis enabled us to determine the range of the most informative indexes among cytokines and microbial cultures that changing their production in order to assess the state of homeostasis in eubiosis and intestinal dysbiosis. It was found that the supernatants of eubiotic cultures of intestinal symbionts exhibited a pronounced ability to inhibit the level of pro-inflammatory cytokines (IFNγ, IL-8) and to stimulate the secretion of anti-inflammatory cytokine (IL-10), whereas the dysbiotic cultures predominantly induced pro-inflammatory cytokines (IL-17, IFNγ, TNFα). In maintaining a uniform balance between pro- and anti-inflammatory cytokines during eubiosis, both associations of microsymbionts (in descending order of factor loads):
Bacteroides
spp. >
E. coli
>
Lactobacillus
spp.), and monocultures (Bifidobacterium spp. and Lactobacillus spp.) made a significant contribution via IL-10 induction. In cases of intestinal dysbiosis, we found an increased number of associations between microsymbionts inducing secretion of pro-inflammatory cytokines was. The pro-inflammatory profile of dysbiotic cultures was determined by the influence on IFNγ production (ranged in descending order of factor loads) of
Bifidobacterium
spp. >
Enterococcus
spp. >
E. coli
>
Lactobacillus
spp. associations, as well as
S. aureus
>
Candida
spp associations. The secretion of IL-17 was influenced by the monoculture of
Clostridium
spp., and by association
C. acnes
>
S. aureus
>
Klebsiella
spp. Monocultures of Bifidobacteria and Escherichia exerted effects upon TNFα production. Thus, during eubiotic state, the normobiota maintains a uniform balance of pro- and anti-inflammatory cytokines, and, in presence of intestinal dysbiosis, a shift in the balance of cytokines towards pro-inflammatory ones may occur due to increased levels of their secretion, an expanded spectrum of cytokines from this group, and increased number of single bacteria and associations of microbial cultures affecting their production.
The environmental determination of indigenous (constantly present) bifidobacteria of the human large intestine is considered in this review. Environmental determination (from the Latin
determinere
, ...“I determine”) is understood as a set of natural phenomena of a habitat (biotope) that determine the role of indigenous microorganisms in the microbiocenosis. Using the symbiotic approach, an attempt is made to identify the environmental conditions for the habitat of bifidobacteria and their physiological effects in the microsymbiocenosis. The features of indigenous bifidobacteria in terms of their nature have been established: evolutionary−genetic (phylogenetic remoteness, genome conservation, metabolic specialization), biochemical (lysozyme resistance, constitutive acetate production), and physiological (microbial “friend−foe” identification, immunoregulation), which are important in adaptation (persistence) and the provision of mutualistic effects and stability of the bifidoflora in the population.
In this study, we determined the levels of cytokine secretory inhibitors and the microbiota biofilms of semen from healthy and infertile subjects. A total of 118 clinical bacterial isolates were ...isolated and tested. Cytokine secretory inhibitors were determined based on the difference in cytokine content between the control and experimental samples of cell-free supernatants of isolated microorganisms. Biofilm formation was studied by determining the adhesion of microorganisms to the surface of a 96-well sterile plate and expressed as the optical density at 630 nm (OD630). Cell-free supernatants of Staphylococcus contained higher levels of secretory inhibitor of cytokines in conditionally healthy than in infertile patients. In contrast, in infertile men, the ability to reduce cytokine levels was more characteristic of Enterococcus and Corynebacterium. Seminal Staphylococcus, Corynebacterium, and Enterococcus isolated from infertile subjects showed a greater ability to form biofilms than the same bacteria isolated from healthy men. Further research is needed on this topic, since it is necessary to determine the relationships between decreased secretory inhibitors of cytokines, production of biofilms by bacteria in semen, and infertility. It is likely that the ability of microorganisms to change the concentration of cytokines and increase the level of biofilm formation in semen may be associated with minimal impairments of fertilizing ability, which are not detected using other methods.
The indigenous human microbiota is a natural storehouse of bifidobacteria strains, the key to our health. Todetermine the indigeneity of symbiont microorganisms, it is proposed to evaluate their ...adaptive potential (lysozyme resistance and biofilm formation), since adaptation is characteristic of all living systems, and in the human body the main adaptation strategy for bacteria is persistence. One of the mechanisms of persistence of indigenous bifidoflora is resistance to the action of host lysozyme, realized through the modification of peptidoglycan – O-acetylation (a wide range of O-cetylate determinants and a family of σ-factors). An important function of indigenous bifidobacteria in the functioning of the human digestive tract is their ability to participate in microbial recognition of “friend or foe”. The results obtained made it possible to open access to a source of beneficial indigenous microorganisms and use them in the selection of cultures for the creation of modern biological products.
A simple accelerated method for determining «self/non-self» microorganisms by using the agglutination reaction (RA) and therapeutic/prophylactic serum (Immunoglobulin complex preparation, lyophilized ...IgG, IgA, IgM immunoglobulins, developed by CSC Immuno-Gem, Moscow) is proposed to test for pathogenic, opportunistic and dominant probiotic Bifidobacteria spp. In parallel, all the microbial cell cultures examined were registered in the databases of Russia-wide and international collections and tested by the intermicrobial “self/non-self” recognition method, previously developed by us. 16 collection strains of various microorganisms were assessed by the RA with relevant therapeutic and prophylactic serum. Biological samples were obtained from the collection bacterial strains of Bifidobacterium bifidum 791, Escherichia coli LEGM-18, Klebsiella pneumoniae 278, Lactobacillus fermentum 90T-C4, Bifidobacterium longum MC-42, Escherichia coli M-17, Shigella sonnei 177b, Shigella flexneri 170, Escherichia coli 157, Staphylococcus aureus 209, Candida albicans 10231 and Salmonella serovar Enteritidis ATCC 10708. In addition, cell cultures obtained from the Museum of the Institute of Cellular and Intracellular Symbiosis UB RAS such as Bifidobacterium longum ICIS-505, Lactobacillus acidophilus ICIS-1127, Bifidobacterium bifidum ICIS-202, Bifidobacterium bifidum ICIS-310 were also included into the study. To assess microbial peptidoglycan foreignness, the intermicrobial “self/non-self” recognition method was also used based on inducing metabolites produced by the “dominant” test strain Bifidobacterium longum MC-42 after pre-incubation with metabolites collected from the studied cell cultures (“associates”) followed by established “dominant-associate” feedback loop. The data were evaluated by assessing change-fold in reproduction (growth/replication) and adaptation (biofilm formation and anti-lysozyme test) of microbial cultures in accordance with the described technique followed by comparing these two methods for intermicrobial “self/non-self” recognition. All the RA data were found to fully agree with those obtained after previous studies by using intermicrobial “self/non-self” recognition method coupled to “dominant-associate” system. Moreover, compared to analogous “intermicrobial recognition” method (5 days), ease of use and test timeframe (24 hours) allow to consider RA attractive for screening studies to select strains for scientific and industrial purposes.
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