Changes in Functional Activity of Neutrophils and Monocytes Isolated from the Peripheral Blood of Women at Different Phases of the Menstrual Cycle.

We studied functional activity of neutrophilic granulocytes and monocytes isolated from the peripheral blood of women during the follicular and luteal phases of the menstrual cycle. It was shown that phagocytic activity of neutrophilic granulocytes increases, their intracellular oxygen-dependent bactericidal activity decreases, and the number of monocyte extracellular traps increases in women in the luteal phase of the menstrual cycle in comparison with the follicular phase.

[FEATURES OF MODIFYING EFFECT OF LOW-INTENSITY ELECTROMAGNETIC RADIATION OF NATURAL AND TECHNOGENIC ORIGIN ON VIABILITY AND FUNCTIONAL STATUS OF NEUTROPHILIC GRANULOCYTES].

AIM: Study effects of microwave electromagnetic radiation (EMR) of the SHF range in vitro on viability and functional status of neutrophilic granulocytes. MATERIALS AND METHODS: Neutrophils of peripheral blood of 30 conditionally healthy donors aged 18 - 20 years were exposed to the effect of broadband (natural, similar to sun radio emission) and, mono-frequency (technogenic, similar to emission of cell phones, PCs, microwave ovens, etc.) EMR with frequency range of 4 - 4.34 GHz, generated by a SHF-generator <>. 16 minutes after the effect, viability of neutrophilic granulocytes, phagocytic activity, lysosome activity ofcells were studied, NBT-reducing ability and cytokine content in supernatant were evaluated. RESULTS: The amount of viable neu- trophils significantly reduced after the effect of modelled EMR with technogenic frequency- temporal structure, functional status of viable cells did not change. Neutrophils retained viability after the effect of EMR of natural origin, increased levels of IL1ß and TNFα was determined in supernatants, functional status of neutrophils of men remained stable, and in women an increase of phagocytic ability and a reduction of production of oxygen radicals was noted. CONCLUSION: The detected reduction of the amount of viable neutrophils under the effect of EMR of technogenic origin could result in disturbances in the system of innate immunity, other homeostasis elements of the human organism and development of pathologic conditions. At the same time, the de- tected effects of EMR of natural origin. open perspectives of use of modelled microwave EMR of SHF range in prophylaxis and clinical medicine.

[EFFECT OF LONGIDAZE ON MICROBIAL LANDSCAPE OF CERVICAL CANAL AND UTERINE CAVITY DURING THERAPY OF WOMEN WITH CHRONIC NONSPECIFIC ENDOMETRITIS AND UTERUS MYOMA].

AIM: Evaluation of microbiological effectiveness of longidaze preparation use during therapy of active non-specific endometritis in women with uterus myoma. MATERIALS AND METHODS: 2 groups of women were formed by a method of random selection. The first was composed of patients with active chronic endometritis, that had received standard etiotropic antibacterial therapy. The second included patients, that had received longidaze in addition to the standard therapy. Therapy effectiveness evaluation was carried out based on the analysis of microbial landscape of cervical canal and uterine cavity 2 months after the therapy during phase I of the menstrual cycle. RESULTS: Non-plasma-coagulating staphylococci, enterococci and anaerobes were established to be present predominately in the examined women of both the first and the second group. CONCLUSION: The application of longidaze in combination with antibacterial therapy results in the most significant improvement of microbial landscape of cervical canal and uterine cavity.

The Role of Neutrophil Granulocyte Ultrastructures in the Formation of Extracellular Traps.

We demonstrated the contribution of cytoskeleton, calcium channels, energy metabolism, and membrane of neutrophils in the formation of neutrophil extracellular traps.

Coherent Fluctuation Nephelometry: A Rapid Method for Urine Screening for Bacterial Contamination.

Express-test by the method of coherent fluctuation nephelometry for urine contamination was carried out on two prototype instruments with standard polystyrene photometric cuvettes. We analyzed 209 and 119 urine samples. Due to high sensitivity of the method, up to 50% negative samples were detected within 10 min by initial opacity and 90% negative samples were detected during 3.5 h by registration of the bacterial growth curves.

[The detection of concentration of DNAase i in seminal fluid.]

The DNAase I is the most studied one among other DNAases. The comparative studies demonstrated that DNAase I from various biological fluids has similar characteristics. The article presents the results of detection of concentration of DNAase I in seminal fluid of conditionally healthy males using technique of enzyme-linked immunosorbent assay. The adaptation of test system DNase ELISA Kit is demonstrated to detect DNAase I in seminal fluid using two modes of calculation of end results.

Oxygen-dependent metabolism of intact and activated neutrophil granulocytes of the saliva.

The effects of phorbol-12-myristate-13-acetate on functional activity and viability of neutrophil granulocytes in the saliva of women at various phases of menstrual cycle were studied using NBT-test and method for detection of neutrophil traps in mucosa secretions. The activator increased the number of neutrophil traps and parameters of neutrophil NBT-test.

[Environmental influences on cellular and humoral immunity in athletes].

The study of seasonal changes in the state of cellular and humoral immunity in trained athletes with predominantly aerobic energy supply of muscle activity, but different different dynamics of the level of physical activity in the annual cycle has been performed. The immune status of athletes, the level of several hormones and interleukins have been established to undergo significant, regular fluctuations training--competitive load throughout the annual cycle that may affect the level of morbidity of athletes.

[The influence of ultrasonic radiation on the factors of antimicrobial resistance in the women of reproductive age presenting with mycoplasma infection of the urogenital tract].

The present clinical immunological study included 90 women of reproductive age presenting with mycoplasma infection of the lower urogenital tract. Low-frequency ultrasonic radiation was shown to successfully correct the disbalance in the system of congenital immunity that manifested itself as marked dysfunction of neutrophil granulocytes. The local application of low-frequency ultrasonic radiation normalized concentration of neutrophil defensins in cervical secretion as well as the number and function of neutrophils themselves.

[The comparative study of indicators of immunity and state of pro- and antioxidant systems in patients with chronic pyelonephritis of single kidney in active and latent stages of disease].

It is established that in patients with chronic pyelonephritis of single kidney the antibacterial defense of organism and balance between pro- and antioxidant systems are disturbed both in active and latent stages of disease. The results testify the need of organization of comprehensive immunologic and biochemical examination to establish a prognosis of complications and evaluation of effectiveness of applied treatment.

[Techniques for detection and role of extracellular neutrophil traps in antimicrobial protection].

New methods are proposed for the measurement of the number of extracellular neutrophil traps in peripheral blood and mucosal secretion. Effect of microbial factors on the ability ofgranulocytes to release nuclear DNA into extracellular space and form extracellular traps is evaluated. Normal microflora species are shown to more actively stimulate build-up of extracellular network. Formation of extracellular neutrophil traps may be a major efficacious mechanism for mucosal antimicrobial protection.

[Impact of thermolabile enterotoxin of Enterobacter cloacae on levels of caspases 3, 7, and 10 under experimental infection].

AIM: To assess the role of thermolabile enterotoxin of Enterobacter cloacae on level of caspases 3, 7, and 10 in experiment. MATERIALS AND METHODS: Observation of apoptosis in mice splenocytes and peritoneal exudate cells. Experimental infection was created by intraperitoneal injection of live bacteria and cultural fluid of E. cloacae producing thermolabile enterotoxin. RESULTS: The study showed that thermolabile enterotoxin of E. cloacae does not have equal effect on apoptosis of studied cells: it slows apoptosis of splenocytes and virtually does not have any influence on peritoneal phagocytes. CONCLUSION: Apoptosis of infected cells is a protective reaction of microorganism to invasion of infectious agent. Cell death leads to rapid elimination of pathogenic agent. Furthermore, cell death by apoptosis compared to necrosis is more favorable for bacteria because it is not induce inflammatory reactions. In our experiments thermolabile enterotoxin of E. cloacae had had antiapoptogenic effect on mice splenocytes that could be a key element in pathogenesis of diseases caused by enterotoxin-producing strains of Enterobacter.

Monitoring of the effect of low-intensity laser radiation with constant pulse generation on neutrophil granulocytes in vitro.

We studied the effect of low-intensity laser radiation with constant pulse generation on bactericidal activity of neutrophilic granulocytes, in particular, on their capacity to form extracellular structures, so-called extracellular neutrophil traps. It was found that exposure to low-intensity laser radiation with constant pulse generation enhanced bactericidal activity of neutrophilic granulocytes, which manifested in the increase of the percent of neutrophils forming extracellular neutrophil traps.

[Analysis of parameters of reproductive tract mucosal immunity in women with chlamydial infection before and after local magnetolaserotherapy].

The objective of the present study was to evaluate the influence of combined treatment with low-intensity laser radiation and magnetic field on neutrophil function in women presenting with Chlamydial infection. Dysfunction of neutrophil granulocytes in these patients was manifest in the first place as the decreased number of phagocytes and the low rate of phagocytosis. It was shown that the concentration of active oxygen species in neutrophils in the patients with Chlamydial infection was significantly smaller than in healthy women. The concurrent application of low-intensity laser radiation and a magnetic field not only stimulated phagocytosis but also increased intracellular production of active oxygen species especially under in vitro conditions. It is concluded that combined treatment with low-intensity laser radiation and magnetic field has beneficial effect on the parameters of mucosal immunity in the reproductive tract of women with Chlamydial infection.

[Influence of Solcotrichovac vaccine on formation of neutrophil extracellular traps].

AIM: To study the influence of Solcotichovac vaccine on formation of neutrophil extracellular traps. MATERIALS AND METHODS: Solcotrichovac vaccine administered in various doses was used in order to assess the formation of neutrophil extracellular traps in peripheral blood of women of childbearing age. RESULTS: Therapeutic dose of Solcotrichovac stimulated the most prominent formation of neutrophil extracellular traps. CONCLUSION: Solcotrichovac vaccine stimulated non-specific arm of mucosal immunity due to formation of neutrophil extracellular traps and it is probable that this mechanism explains the effect of this vaccine.

[Neutrophil extracellular traps: method of detection and assessment of bacterial trapping efficacy].

AIM: To develop express method for detection of neutrophil extracellular traps represented by extracellular strands of neutrophils' DNA. MATERIALS AND METHODS: Acridine orange staining was used for visualization of neutrophil exracellular traps, which allows to quickly reveal the forming structures and to quantify this phenomenon. RESULTS: Performed studies showed that after 30-min of in vitro interaction of Escherichia coli, Staphylococcus aureus, lactobacteria, bifidobacteria and latex particles with neutrophils, the latter produces extracellular net-like structures, which were well visualized after staining of fixed preparations with acridine orange. Such structures are able to trap latex particles and bacteria more efficiently than live cell. Additionally, representatives of normal flora (bifidobacteria) actively stimulate production of traps. CONCLUSION: Production of neutrophil extracellular traps could be one of the main and effective mechanisms in antimicrobial defense of mucosal surfaces.

[Effect of Enterobacter cloacae thermolabile enterotoxin on immune system of mice].

AIM: To study the effect of Enterobactercloacae thermolabile enterotoxin (LT-enterotoxin) on different arms of immune system of mice. MATERIALS AND METHODS: Proprietary clinical strain E. cloacae 258 was used and intraperitoneal inoculation of outbred white mice with LT-enterotoxin-producing E. cloacae comprised experimental model. Phagocytic and lysosomal activity of peritoneal macrophages as well as level of APC to sheep erythrocytes were measured, spontaneous and induced NBT tests were also performed. RESULTS: Pleiotropic immunotoxic effect of LT-enterotoxin of studied bacteria was established, which characterized by suppres- sion of antigen-presenting and antigen-processing functions of macrophages, stimulation of mitotic activity of lymphocytes, decrease of activity of hexose monophosphate shunt enzymes in peritoneal macrophages as well as abnormalities in formation of specific B-lymphocytes. CONCLUSION: Obtained data allowed to reveal separate steps in pathogenesis of infectious process caused by LT-enterotoxin-producing bacteria from Enterobacter genus.

[Study of effect of thermolabile enterotoxin of Enterobacter cloacae on expression of cytokine genes in mice].

AIM: To study the role of production of several cytokines by macrophages and neutrophils in experiment. MATERIALS AND METHODS: Production of cytokines--IL-2, 4, 6, 10, 12, 15 as well as interferon gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), transforming growth factor beta (TGF-beta) by macrophages and neutrophils during experiment on mice inoculated with enterotoxigenic strain of Enterobacter cloacae was measured. Expression of cytokines was determined by reverse transcription PCR and dot-hybridization on the basis of specially synthesized nucleotide sequences. RESULTS: In macrophages of experimental animals inoculated with enterotoxigenic strain of E. cloacae increased production of TGF-beta, IFN-gamma and decreased level of TNF-alpha were revealed. Transcription of mRNAs of IL-15 and IL-6 was detected in macrophages and neutrophils of experimental animals at 24 h after inoculation. Transcription of mRNAs of IL-2, IL-4 and IL-10 was detected neither in macrophages nor in neutrophils. CONCLUSION: Thermolabile enterotoxin of E. cloacae renders negative effect on IFN-gamma, promotes enhanced expression of TGF-beta and suppresses production of IL-2, IL-4, IL- 10. Absence of expression of key cytokines suggests that effect of thermolabile enterotoxin of Enterobacter bacteria excludes it from cytokine regulation of immune response during infectious process.

[Interaction of neutrophils and different bacterial agents].

Neutrophils migrate on the epithelial surface and interact with microflora of mucous membrane, then activate, realize their effect or functions and destroyed as a result of necrosis or apoptosis. We determined the influence of Staphylococcus aureus, Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. on viability of neutrophils. It was revealed that influence of different bacterial factors on the process of neutrophils' apoptosis is not equal: resident bacteria of women genital tract mucous membrane (Lactobacillus spp. and Bifidobacterium spp.) have powerful apoptogenic effect on neutrophils, unlike the facultative microorganisms (S. aureus and E. coli) which have lesser activity.