ADRENERGIC REGULATION OF IMMUNE RESPONSE UNDER EXPERIMENTAL THYROTOXICOSIS IN RATS

It was established that a changes of antibody production and reaction of delayed—type hypersensitivity at the local immune response to thymus-dependent antigen were opposite in rats with varying severity of experimental thyrotoxicosis — stimulation under a mild form of thyrotoxicosis and depression under more severe form. The increase in glucocorticoid level may be involved in the mechanism of immunosuppression under severe form thyrotoxicosis. The administration of agonist or antagonist of beta-adrenoceptors to animals with thyrotoxicosis led to the significant modification of changes of an immune response parameters, that confirms the participation of an increase in the sensitivity to beta-adrenergic regulation of immune system cells in immunomodulation under thyrotoxicosis

Study of the immunoglobulin and oxidized protein content of semen under infertility

Among the causes of male infertility, enough attention is paid to oxidative stress, which in turn is a pathogenetic link in the inflammatory process. However, there is practically no information on the content of oxidized modified proteins in the semen, which makes it difficult to study the pathogenesis of diseases of the male reproductive system. In part, protein oxidation may be due to the production of reactive oxygen species by microorganisms, both directly and indirectly through the activation of immune system cells. The aim of the research was to study the level of oxidized modified proteins and changes in immunoglobulin concentrations in the semen under bacteriospermia. A study was made of the ejaculate of 48 men who applied to the clinic for infertility in marriage. The comparison group consisted of 32 practically healthy men who had no growth of microorganisms in the ejaculate samples. When conducting bacteriological analysis, the studied samples were diluted 10 times and used the generally accepted method. The concentration of albumin, immunoglobulins A, M, G, E was determined in the spermatic fluid. The oxidative modification of proteins was evaluated in the reaction with 2,4-dinitrophenylhydrazine. The concentration of oxidized proteins was expressed in nmol/mg of the total protein of the studied biological fluid. The biuret method was used to determine the protein concentration. Statistical analysis of the results was performed using descriptive statistics and Student's t-test for paired data. The concentration of protein in the seminal fluid did not differ significantly among the studied groups. The albumin concentration (16.96±1.28 mg/mL) was statistically significantly lower in the absence of microorganism growth than in bacteriospermia. With bacteriospermia, a decrease in the concentration of IgM and IgA and an increase in the level of IgG were noted. The degree of protein oxidation is maximum when enterobacteria are isolated from seminal fluid. Thus, during the studies it was found that, despite the absence of a clinic, with asymptomatic bacteriospermia, the secretion of immunoglobulins G into the semen is observed. The accumulation of oxidized proteins in the seminal fluid in bacteriospermia has been shown

Influence of polyamines of bacterial origin on the production of key cytokines in the culture of human mononuclear leukocytes

To date, participation of biogenic polyamines has been studied in details, with respect to regulation of microbial gene expression, interrelations between bacteria, development of their persistence state. Opportunity of their use as markers of human pathological conditions is being actively evaluated. The aim of our study was to assess the effect of bacterial diamines, i.e., cadaverine and putrescine, upon production of key cytokines (IFNγ and IL-4) in the culture of human mononuclear leukocytes. We studied leukocytes of peripheral venous blood obtained from 18 healthy male volunteers (mean age 24.0±0.6 years). The leukocytes were isolated by means of gradient centrifugation using a Ficoll-Verografin mixture. For the cultivation of lymphocytes, a micro-method and plastic round-bottom 96-well plates were used. Concanavalin A at a concentration of 5 μg/ml was used as a T cell mitogen. Polyamines were used at final concentrations of 5, 25, 50, 75 and 100 μM/L. The cultivation was carried out in humidified atmosphere with 5% CO2, at 37 °C for 72 hours. At the end of incubation, the culture medium was collected and frozen for subsequent quantitative enzyme immunoassays of cytokine concentrations (IFNγ and IL-4) (Russia). Viable cells were counted using Goryaev chamber after staining with 0.1% trypan blue solution. Statistical analysis was performed using Student’s t test or Mann–Whitney test. Addition of cadaverine at all concentrations reduced IFNγ production in the culture of mitogen-activated cells. When culturing leukocytes supplied with putrescine (5 to 50 μM/L), a dose-dependent decrease of IFNγ was observed. Upon further increase of putrescine concentrations, the IFNγ production is restored to the values of the control samples. Direct toxic effect of polyamines upon the cells was revealed. Both the diamines, at the doses of 50, 75, and 100 μM/L, caused increase of IL-4 production by the mitogen-activated cells. Such changes can be associated both with direct cytotoxic effect of cadaverine and putrescine, being mediated by changes of some metabolic pathways. In addition, the effects of polyamines upon monocytes present in culture can include their anti-inflammatory state, e.c., an increased IL-4 production. In general, cadaverine and putrescine, produced by microorganisms of various taxonomic groups, regulate the effectiveness of compensatory-adaptive reactions that ensure adaptation of microbial populations to changing or unfavorable environmental conditions

TECHNICAL ASPECTS OF EVALUATING CYTOKINE PROFILE AND LEUKOCYTE REACTION IN BILE DUCT OBTURATION OF DIFFERENT ORIGIN

The study was carried out to assess the prospects for the combined use of leukocyte and cytokine indices in the bile duct obstruction of different genesis.We have performed a study of the patients with mechanical jaundice, either due to tumor (14 persons), or non-tumor genesis (16 persons). Concentrations of cytokines (IL-1, IL-4, IL-8, TNFα) were determined in blood serum before surgical intervention. The total activity of endogenous inflammatory mediators in the serum of patients was estimated as an index of inflammatory activity (IVA), which was calculated using the following formula: IVA = (IL-1 + TNFα)/IL-4. In addition, a number of leukocyte indices were calculated. The cellular component in the index estimation makes it possible to clarify the intensity of the inflammatory reaction, and to suggest the direction of the functional changes among the  immunocompetent cells.It was shown that IVA in presence of malignancy is significantly higher than in benign conditions. Overactivation of immune response in tumors may be caused by a significant predominance of pro-nflammatory cytokines. Moreover, in cases of malignant tumor growth, both with or without cholangitis, the index scores of humoral and cellular components let us suggest the some problems with regeneration, due to pro-inflammatory domination of immune response, which does not ensure activation of the corresponding reactions. It has been established that the leukocyte intoxication index (LII) is increased,in patients with a combination of malignant disorder and cholangitis, and the immunoreactivity index (MDI) is lowered. In absence of cholangitis, the opposite pattern is observed, when LII is slightly lowered and MDI is elevated. Correlation analysis for the patients with cholangitis revealed a high association of cytokine imbalance and endogenous intoxication. A complex of cytokine and leukocyte indices is promising, when each of them reflects a certain part of the process, and their combination gives an integral picture of the pathology and allows to predict development of the disease. Such an approach, seems to increase the diagnostic significance of these immunological indices, primarily, the cytokine production, and may be used when predicting currence of complications in the surgical treatment of the disease

Experimental evaluation of polymer construction materials: effects of acrylic resin and a fiberglass-based composite upon cytokine production

Polymethyl methacrylate-based acrylic resin is commonly used in current dental practice as an underlying material for fabrication of overdenture restorations. To increase service life of laminar overdentures, we have developed a technique to fabricate a novel combined base for a full overdenture using a fiberglass-based composite. It makes sense to evaluate probable hazards of inflammatory process which could be activated when using structural polymer materials and composites in dental practice. Our study aimed for assessment of changes in production of key cytokines (interferon-y and interleukin-4) by ex vivo incubated human mononuclear leukocytes in the presence of acrylic resin and fiberglass-based composite.The experiments dealt with peripheral venous blood leukocytes obtained from 13 apparently healthy male volunteers (mean age = 24 years). The leukocytes were isolated from heparinized blood by gradient centrifugation. The cells were cultured in plastic round-bottom 96-well plates, in moist atmosphere with 5% CO2 at 37 °C for 72 h. Following the incubation, the culture supernates were collected and frozen for further determination of cytokine concentration using ELISA reagent kits for interferon-y and interleukin-4 measurement (Vector-Best, Russia). The samples of two structural materials were tested in the bioassays: the specimens of acrylic resin, Ftorax were compression-moulded by hot polymerization in prefabricated casts; the specimens of a composite, Trinia, were computer-milled. Glass specimens of similar shape and size were used as references. The statistical analysis used a software package, Statistica 7.0. Significance of the differences was evaluated using the Student's test and the Mann-Whitney test. When testing the statistical hypotheses, the significance level (p) was taken to be 0.05.Significant decrease of IFNγ production was revealed in presence of Trinia than in the samples with glass and with the acrylic specimens, whereas cell viability counts did not differ from the blank values. There was no statistical differences in IL-4 production between the samples with the polymer materials and the glass. When estimating individual stimulation indexes, the materials used in this research were found to showed a pronounced stimulatory effect with peripheral blood lymphocytes from only one volunteer. These findings indicate that Trinia triggers anti-inflammatory activity of leukocytes, whereas IFNγ production level is somewhat decreased, and IL-4 production remains unchanged.Thus, the research assessed the method for personalized evaluation of reactivity of prosthodontic structural polymer materials. Absence of increase in lymphocytic production of key cytokines can be regarded as a hopeful sign which indicates that inflammatory process is not activated when Trinia is used in overdenture bases

DYNAMICS OF SERUM IMMUNOGLOBULINS IN ONCOLOGICAL PATIENTS DURING PERIOPERATIVE PERIOD

The aim of the study was to investigate the dynamics of serum immunoglobulins from oncological patients in the perioperative period. As a result of investigations, it was established that already at the initial stage of the intraoperative period the IgM concentration was elevated in patients' sera, and persisted up to 3 days after surgical intervention, while exceeding the values of the control group by 30—80 %. At the same time, no significant changes were detected in IgA and IgG levels of examined patients in the perioperative period. The obtained data confirm the expediency of carrying out the monitoring of immunoglobulin levels in oncological patients both in intra-, and postoperative periods and can be used for the assessment of patient's immune status and in the choice of immunocorrecting therapy strategy

Formation of biofilm by conditionally pathogenic microorganisms on metal and polymeric brackets

Most oral microorganisms have the property of adhesion to hard tissues, thus forming a biofilm on them, which serves as the foundation for plaque. With an increase in the area of potential bacterial film formation by long-existing dental structures in the oral cavity, the sedimentation rate and the amount of plaque increase. The analysis of biofilm formation by the main bacteria from the ATCC collection on brackets made of composite polymer, as well as stainless steel, was carried out. A high level of biofilm formation by S. aureus and C. albicans strains on metal and polymer braces has been revealed, which is of interest for the development of new methods for removing plaque, including from the surface of dental constructions.Большинство микроорганизмов ротовой полости обладает свойством адгезии к твердым тканям, образуя при этом на них биопленку, которая выполняет функцию фундамента для зубного налета. При увеличении площади потенциального образования бактериальной пленки длительносуществующими в ротовой полости стоматологическими конструкциями повышается скорость оседания и количество налета. Проведен анализ формирования биопленки основными бактериями из коллекции АТСС на брекетах из композитного полимера, а также из нержавеющей стали. Выявлен высокий уровень формирования биопленки штаммами S. aureus и C. albicans на металлических и полимерных брекетах, что представляет интерес для разработки новых методик удаления зубного налета, в том числе и с поверхности стоматологических конструкций

The Microbiota Continuum along the Reproductive Tract in Women with Infertility

Introduction. At present, the question of commensal, including opportunistic, microflora participation in  infertility development remains debatable. In a number of studies, the translocation of the vaginal microflora into the endometrial tissue is considered as a factor contributing to inflammation development. In addition, the connection of some reproductive losses with the persistence of certain conditionally pathogenic microorganisms is shown. Today, to solve this issue, molecular genetic research methods are being actively introduced that surpass the routine cultivation techniques in a number of positions.The aim of the study was to assess the taxonomic diversity of microorganisms in the vaginal biotope with infertility.Material and methods. For the study samples of vaginal contents obtained from the posterior vaginal fornix of 15 women, consisting of barren marriage, were used. A metagenomic study of 16S ribosomal RNA samples was carried out on the Illumina MiSeq platform, using the MiSeq Reagent Kits v3 kit (600-Cycle Kit), as recommended by the manufacturer. Libraries for sequencing plots of the V3-V4 gene of the 16S ribosomal RNA were prepared according to the 16S Metagenomic Sequencing Library Preparation Illumina. In bioinformatics assessment, Kraken Metagenomics version 2.0.0 software for metagenomic analysis (classifier of reads — short nucleotide sequences) was used using a standard database.Results. It was shown that the occurrence of representatives of the Lactobacillaceae family in the vaginal biotope varies from 12 to 84%. The genus Lactobacillus with the dominance of L. jensenii, L. delbrueckii and L. amylolyticus occupied the leading position among the members of the family. In all samples, Moraxella spp. was found in large numbers, with M. osloensis leading among the representatives of the entire community. In half of the cases, the joint presence of M. osloensis and G. vaginalis was revealed.Conclusion. It has been established that in infertility in the vaginal microbiota the number of representatives of the genus Lactobacillus is significantly reduced, and there is also a change in the leading species to L. jensenii, whose functional activity does not fully ensure the colonization resistance of the vaginal biotope, allowing for excessive reproduction of oppotrunistic microorganisms, in particular, M. osloensis