Search

Objective: To investigate the role of dopamine receptor D 1 DR and D 2 DR in the production of cytokines interleukin-6 (IL-6) and IL-1β by monocytes and macrophages in patients with relapsing-remitting multiple sclerosis (MS)., Material and Methods: Ten patients with relapsing-remitting MS and 10 healthy subjects were examined. The level of IL-6 and IL-1β production was assessed in culture supernatants obtained from CD14 + monocytes or macrophages stimulated with interferon-γ (IFN-γ) and lipopolysaccharide (LPS). To study the role of dopamine receptors in the regulation of CD14 + monocytes or macrophages, samples of cells were incubated in the presence of specific D 1 DR or D 2 DR antagonists, after which IFN-γ/LPS were added to the cultures. Levels of cytokines in culture supernatants were measured by enzyme-linked immunosorbent assay., Results: The production of IL-6 and IL-1β by CD14 + monocytes and macrophages was comparable between the groups. Blockade of D 1 DR suppressed cytokine production by CD14 + monocytes and macrophages in both groups. In contrast, blockade of D 2 DR increased the production of cytokines by CD14 + monocytes and did not affect cytokine production by macrophages in both groups., Conclusions: Targeting of dopaminergic receptors could be considered as an additional mechanism of immunomodulation in MS with both pro- and anti-inflammatory effects on cells of the innate immune system.

Limbal stem cell deficiency (LSCD) is one of the leading factors negatively affecting the success of keratoplasty, and its treatment remains an urgent problem in ophthalmology. With the development of regenerative medicine, one of the promising approaches is the transplantation of tissue-engineered constructs from cultured limbal stem cells (LSCs) in biopolymer carriers., Purpose: This study was conducted to develop an experimental model of LSCD and evaluate the effectiveness of transplantation of a tissue-engineered construct consisting of cultured cells containing a population of LSCs and a collagen carrier., Material and Methods: The study was performed on 12 rabbits and included several stages. At the first stage, the physiological effects of collagen matrix implantation into the limbal zone were studied. At the second stage, tissue-engineered constructs consisting of LSCs on a collagen matrix were formed and their effect on the regeneration processes in the experimental LSCD model was analyzed. The animals were divided into 2 groups: surgical treatment (transplantation of the tissue-engineered construct) was used in the experimental group, and conservative treatment was used in the control group. Slit-lamp biomicroscopy with photo-registration, fluorescein corneal staining, optical coherence tomography of the anterior segment of the eye, and impression cytology were used to assess the results., Results: No side reactions were observed after implantation of the collagen matrix into the limbal zone. One month after surgical treatment of the LSCD model in the experimental group, complete epithelization with minor manifestations of epitheliopathy was observed. In the control group, erosion of the corneal epithelium was noted. The time of corneal epithelization in the experimental and control groups was 9.2±2.95 and 46.20±12.07 days, respectively ( p =0.139). According to the data of impression cytology, in the experimental group there were no goblet cells in the central part of the cornea, which indicates the restoration of corneal type epithelial cells, in contrast to the control group., Conclusion: Transplantation of a tissue-engineered construct from cultured limbal cells on a collagen membrane should be considered as a promising method for the treatment of limbal stem cell deficiency.

Osteoarthritis is a widespread age-related disease, that has no effective targeted therapy. In this regard, bioengineering methods are being actively developed that can stimulate the restoration of cartilage tissue. These methods include chondrogenic differentiation of stem cells, which is stimulated by various biomolecules, including short peptides and polypeptide complexes. It was studied the effect of the cartilage polypeptide complex (CPC) and AED peptide on gene expression and protein synthesis of chondrogenic differentiation - SOX9, aggrecan, type II collagen and COMP - in human mesenchymal stem cell (MSC) during replicative aging. AED peptide at the concentration of 200 ng/ml activates gene expression and protein synthesis during aging of MSCs. CPC has the same effect in the concentration 2000 ng/ml. These data indicate the stimulating effect of studied peptides on regulation of chondrogenesis and open up prospects for further investigation of their effectiveness in osteoarthritis models.

Determination the activity of the genes of sirtuin-1, hyaluronidase, TGF-β cytokine, calreticulin in the process of replicative aging of human fibroblasts in vitro and the effect of hyaluronan preparations with gold nanoparticles on the activity of replicative cell aging. Compared the expression of proteins of the studied genes using specific markers at 7 and 14 passages of cultivation of fibroblasts isolated from human skin, without drugs and in the presence of drugs in the growth medium. This work shows a decrease in the activity of the sirtuin 1 gene and an increase in the expression of hyaluronidase in the process of replicative aging of human fibroblasts. Found a means of slowing down replicative aging by activating the SIRT-1 gene and reducing the activity of hyaluronidase in action in the growth medium of hyaluronan preparations with gold nanoparticles. The discussed variants of cell transitions to the pathological state, caused by replicative aging and the mechanisms of slowing down the replicative aging of human fibroblasts.

The article presents results obtained on several types of model cultures. Primary cultures of corneal epithelial cells and fibroblast-like cells from the limbus were obtained from the tissues of the anterior eye segment. Tests were performed on these cultures to determine the cytotoxicity of antiglaucoma drugs and to trial a protection method. In addition, a comparative analysis of the regenerative potential of various blood derivatives was carried out. An in vitro culture of endotheliocytes showed a correlation between the degree of damaging effects of antiglaucoma drugs of various groups and the content of the preservative benzalkonium chloride in their composition. Corneal keratocytes were used to create a cell sheet reflecting the main structural features of the stroma and suitable for biomechanical tests. The antifibrotic activity of the drugs was tested on the culture of fibroblasts from the nasolacrimal duct. The conducted studies demonstrate that cell cultures can be used as an informative model object for researching the pathogenesis of ophthalmic diseases and for testing drugs.

It was shown, that genotoxic stress can trigger endothelial disfunction and atherosclerosis, but the molecular genetic mechanisms of this process are poorly investigated. At the same time, inflammation also plays the important role in atherogenesis. This study aimed access of inflammatory marker expression in the endothelial cells exposed to alkylating mutagen mitomycin C (MMC). Primary human coronary (HCAEC) and internal thoracic artery endothelial cells (HITAEC) exposed to 500 ng/ml MMC (experimental group) and 0.9% NaCl (control) were used in this research. A gene expression profile was evaluated by quantitative reverse transcription PCR after 6 h exposure of endothelial cells to MMC (or 0.9% NaCl) followed by subsequent 24 h incubation in the mutagen-free cell growth media. The cytokine profile of endotheliocytes was studied by dot blotting. We found that MIF, IL-8, MCP-1, IP-10 and PDGFB were upregulated both in HCAEC and HITAEC, while MIP-1β release remained unchanged. TIMP-2 was upregulated in HCAEC but not in HITAEC. sTNF RI was expressed only in HCAEC. According to gene expression analysis, HCAEC exposed to MMC are characterized by the increased mRNA level of IL-8, MCP-1 and IP-10; decreased expression of TIMP-2 and no differences in the expression of MIF, MIP-1β and PDGFB compared to the control. In HITAEC, increased mRNA level of IL-8 and IP-10; decreased expression of MIF and TIMP-2, no differences in the expression of MCP-1, MIP-1β and PDGFB was shown. TNF-RI expression was not detected in both cell lines. Thus, genotoxic stress in endothelial cells induced by MMC leads to differential inflammatory response that can trigger endothelial dysfunction.

The expression of DNA repair (DDB1, ERCC4, ERCC5), leukocyte adhesion (VCAM1, ICAM1, SELE, SELP), endothelial mechanotransduction (KLF4), endothelial differentiation (PECAM1, CDH5, CD34, NOS3), endothelial-to-mesenchymal transition (SNAI1, SNAI2, TWIST1, GATA4, ZEB1, CDH2), scavenger receptors (LOX1, SCARF1, CD36, LDLR, VLDR), antioxidant system (PXDN, CAT, SOD1) and transcription factor (HEY2) genes in primary human coronary (HCAEC) and internal thoracic (HITAEC) arteries endothelial cells exposed to alkylating mutagen mitomycin C (MMC) was studied at two time points - after 6 h of incubation with MMC and after 6 h of the genotoxic load followed by 24 h of incubation in pure culture medium using the quantitative PCR. Immediately after MMC exposure, in the exposed HCAEC and HITAEC a decreased expression of almost all studied genes was noted excepted SNAI, which demonstrated a 4-told increase in its expression compared to the unexposed control. Elimination of MMC from the cultures, an increased expression of the VCAM1, ICAM1, SELE, SNAI2, KLF4 genes and a decreased the mRNA level of the PECAM1, CDH5, CD34, ZEB1, CAT, PXDN genes were observed in both cell lines. In addition, HITAEC cells were characterized by a decreased expression of the SOD1, SCARF1, CD36 genes and an increased expression of the SNAI1 and TWIST1 genes; in HCAEC, an increased mRNA level of the LDLR and VLDLR genes was noted. Thus, MMC-induced genotoxic stress is associated with the endothelial dysfunction.

Effective and safe antiseptic eye preparations are necessary for prevention and treatment of infectious and inflammatory eye diseases., Purpose: in vitro evaluation of the effect of antiseptic eye drops on corneal and conjunctival epithelial cells., Material and Methods: Antiseptic eye drops «Bactavit», «Vitabact» and «Ocomistin» were the object of the study. Immortalized human corneal epithelial cell lines (HCE) and human conjunctiva (Chang Conjunctiva, Clone 1-5c-4) were used as the test systems. The viability of the cells was assessed by their metabolic activity and morphology using the MTT test and phase-contrast microscopy., Results: Antiseptic eye drops belonging to different groups of chemical compounds induced cytotoxic effects on the cells of corneal epithelium (HCE) and human conjunctiva (Chang Conjunctiva, Clone 1-5c-4) of varying degrees, leading to morphological and functional changes in those cells., Conclusion: The study confirms the possibility of using cultured cells for the in vitro comparative assessment of the cytotoxic effect of antiseptic ophthalmic agents.

A study was made of the effect that mitomycin C (MitC) treatment of stromal layers of NIH 3T3 cells expressing Jagged1, a ligand of the Notch receptor, exerts on the growth of hematopoietic Lin(-) mouse bone marrow cells in a co-culture system. MitC treatment of stromal cells significantly increased the number of hematopoietic cells and the frequency of colony-forming cells in stromal co-cultures. Transcriptome analysis of control and MitC-treated stromal cell samples was performed by differential RNA sequencing, and genes downregulated by MitC treatment were predominantly associated with the control of cell proliferation, the cell cycle, chromosome segregation, and DNA metabolism. Induction of key hematopoietic cytokines by MitC was not detected by the transcriptome analysis and was therefore not a main factor in the activation of hematopoiesis on the treated stroma. At the same time, the set of the genes most strongly upregulated by MitC treatment is enriched in the genes for cytokines, growth factors, and cell surface proteins, which presumably contribute to enhanced hematopoiesis support on the MitC-treated stroma. Products of some of these genes have been implicated in expansion of hematopoietic stem/progenitor cells in vitro or in vivo.

Recently, SGI-1027, a well-known inhibitor of DNA-methyl transferases (DNMTs), was reported to effectively reduce formation of pathogenic PrP^(Sc) in prion-infected cells. Herein, we confirm the elimination of PrP^(Sc) in chronic wasting disease (CWD) prion-infected neurons by SGI-1027, and pinpoint the binding region of human prion protein to SGI-1027. SGI-1027 is broadly functional against various prion disease types, including human prions. Previously, the inhibitory effects of SGI-1027 on DNMT function is well tested in various cell culture models. While neither treatment with a DNMTs enhancer S-adenosyl-L-methionine (SAM), nor with their inhibitor, 5-azacytidine, prevented PrP^(Sc) propagation, SGI-1027 did. Our study suggest that the anti-prion effects of SGI-1027 are a result of its direct interaction with PrP^(C), which effectively interferes with the pathogenic conformational change of PrP^(C) to PrP^(Sc). We conclude that SGI-1027 driven suppression of pathogenic PrP^(Sc) is independent of DNMT.

In the model of induced neuronal resistance to the toxic effect of glutamate (deprivation of trophic factors), exosome secretion is demonstrated. Exosomes are secreted at the development of resistance during deprivation and at the first 24 h after preconditioning, as was shown by dot blot of extracellular fluid using anti-CD63 antibody. The autophagy inhibitor bafilomycin (0.01 μM) significantly reduces the quantity of the secreted exosomes at the stage of autophagy induction and at 24 h after induction. At the same time, inhibition of autophagy during the deprivation of trophic factors prevents the development of resistance, but inhibition of autophagy during the first 24 h after deprivation does not affect the development of resistance. We suggest that the long-term effects of preconditioning may be mediated by exosome secretion.

Neurogenesis is a complex process which governs embryonic brain development and is importants for brain plasticity throughout the whole life. Postnatal neurogenesis occurs in neurogenic niches that regulate the processes of proliferation and differentiation of stem and progenitor cells under the action of stimuli that trigger the mechanisms of neuroplasticity. Cells of glial and endothelial origin are the key regulators of neurogenesis. It is known that physiological neurogeneses is crucial for memory formation, whereas reparative neurogenesis provides partial repair of altered brain structure and compensation of neurological deficits caused by brain injury. Dysregulation of neurogenesis is a characteristics of various neurodevelopmental and neurodegenerative diseases, particularly, Alzheimer's disease which is very important medical and social problem. In the in vitro model of the neurogenic niche using hippocampal neurospheres as a source of stem/progenitor cells and astrocytes, we studied effects of astrocyte activation on the expression of markers of different stages of cell proliferation and differentiation. We found that aberrant mechanisms of development of stem and progenitor cells, caused by the beta-amyloid (Aβ1-42), can be partially restored by targeted activation of GFAP-expressing cells in the neurogenic niche.

Secretion of 21 cytokines, chemokines and growth factors (LIF, SCF, SDF-1a, SCGF-b, M-CSF, MCP-3, MIF, MIG, TRAIL, GRO-a; IL-1a, IL-2ra, IL-3, IL-12(p40), IL-16, IL-18, HGF, TNF-b, b-NGF, IFN-a2, CTACK) has been studied in vitro in the culture of human adipose-derived multipotent mesenchymal stromal cells (hAMMSCs) in conditions of its osteogenic differentiation caused by 14-day contact with calcium phosphate (CP) surface with different roughness. Bilateral X-ray amorphous CP coatings were prepared on the samples of commercially pure titanium in the anodal regime using a micro-arc method. An aqueous solution prepared from 20 wt% phosphoric acid, 6 wt% dissolved hydrohyapatite nanopowder (particle diameter 10-30 nm with single agglomerates up to 100 nm), and 9 wt% dissolved calcium carbonate was used to obtain CP coating. hAMMSCs isolated from lipoaspirate were co-cultured after 4 passages with the CP-coated samples at final concentration of 1.5´105 viable karyocytes per 1.5 mL of standard nutrition medium (without osteogenic stimulators) for 14 days (a determination of [CD45,34,14,20], CD73, CD90 и CD105 cell immunophenotype; an analysis of secretory activity) and 21 days (alizarin red S staining of culture) with medium replacement every 3-4 days. Under conditions of in vitro contact with rough CP coating hAMMSCs differentiated into osteoblasts synthesizing the mineralized bone matrix; this was accompanied by 2-3-fold increasing ratio of [CD45,34,14,20]+ hemopoietic cells. The following humoral factors of hemopoietic niches acted as the signal molecules escalating in vitro the hemopoietic base in 14 days of differentiating three-dimensional culture of hAMMSCs: either leukemia inhibitory factor (LIF) and stem cell factor (SCF) cytokines under mean index of CP roughness Ra=2.4-2.6 mm or stromal derived factor-1 (SDF-1a, CXCL12 chemokine) under Ra=3.1-4.4 mm.

We studied the effect of the native (non-recombinant) alpha-fetoprotein (AFP) on differentiation, proliferation, and cytokine profile of activated helper T cells 17 (Th17). The object of the study was a culture of isolated by immunomagnetic separation helper T cells (CD4+), induced into the Th17 phenotype by using TCR-activator and proinflammatory cytokines (IL-1β and IL-6). AFP had not significant effect on the frequency of Th17 cells (ROR-γτ+) in the helper T cell culture, and did not affect proliferation of these cells, as measured by Ki-67 expression. Evaluation of the cytokine profile of culture supernatants by using the Luminex xMAP technology, revealed that AFP did not affect the levels of IL-4, IL-5, IL-7, IL-8, IL-10, IL-17, IFN-γ and TNF-α, but at concentrations of 50 IU/ml and 100 IU/ml it increased IL-2 production by activated helper T cells. At the same time, AFP suppressed the synthesis of G-CSF and GM-CSF (10 IU/ml), but stimulated the production of CCL4/MIP-1β (100 IU/ml) and CCL2/MCP-1 chemokines (10 IU/ml and 50 IU/ml).

The purpose of the study was to investigate the features of expression and adiponectin content in the adipocyte culture of subcutaneous, epicardial, and perivascular adipose tissue and the effect of various doses of rosuvastatin on these processes. 29 patients with coronary artery disease were examined. Adipocytes were isolated from the samples of SAT, EAT and PVAT which were taken during coronary artery bypass surgery, followed by cultivation in the presence of rosuvastatin and evaluation of gene expression and adiponectin concentration. Adipocytes SAT, EAT and PVAT differed in the level of adiponectin secretion and expression of its gene. On day 1 of cultivation the expression of the adiponectin gene in the EAT was 2.3 times lower than in the PVAT. On day 2 of cultivation the expression of the adiponectin gene was reduced both in the EAT and the PVAT as compared to the SAT. When rosuvastatin was added at a concentration of 1 mmol/L, adiponectin gene expression in PVAT was higher than when rosuvastatin was added at a concentration of 5 mmol/L, in the adipocyte culture of SAT effect was opposite. Thus, the adipocytes of EZhT and, to a greater extent, PAS, can be a therapeutic target for statins in the case of the pathological activation of adipose tissue.

Gerontocosmetology is the rapid developing knowledge area that has a very large applied meaning. Herewith a lot of information about skin aging and geroprotectors for skin rejuvenation hasn't a scientific background. Thus, understanding the fundamental mechanisms of skin aging becomes the actual task of molecular gerontology. Skin fibroblasts are the polyfunctional cell population that synthesize a number of biologically active substances and participate in maintaining of extracellular matrix homeostasis, skin hydratation and endocrine and immune function. In the review genetic (accumulation of nuclear and mitochondrial DNA mistakes) and epigenetic factors of skin fibroblasts aging are described. Role of AP-1, NF-κB, c-jun, CCN1, TGF-β, TNF-α, MMP-1, MMP-3, MMP-8, MMP-9 and glycation in skin fibroblasts aging are discussed. There are some data about decreasing of skin fibroblasts ability to migration and synthesis of paxillins and aquaporin-3 (AQP3) during aging. Role of hormonal regulation in skin fibroblasts aging are described. Geroprotective action of melatonin to skin fibroblasts are showed. Reviewed molecular-cellular aspects of skin fibroblasts aging can be take into consideration for scientific background of using of cosmetic products for retarding of skin aging rate.

Modeling of tuberculosis infection is carried out in order to clarify various aspects of the tuberculosis pathogenesis, as well as the testing of new anti-tuberculosis drugs. The characteristic of in vitro models (n = 16) for Mycobacterium tuberculosis dormant state and in vivo models (n = 14) for the latent tuberculosis infection involving several animal species published to date are presented in this review. A brief description of the models and the results obtained by the authors are presented. The analysis of the published data reflects the list of methodological procedures that allow researchers to study the mechanism of the transition of M. tuberculosis cells to a dormant state and reverse to metabolically active state, as well as the process of conversion of active tuberculosis infection to a latent tuberculosis and reactivation., Competing Interests: The authors declare no conflict of interest.

Lipopolysaccharides (LPS), components of the cell wall of gram-negative bacteria, activate neutrophils that trigger pathological processes, including gram-negative sepsis. LPS inhibit spontaneous apoptosis of neutrophils that leads to inflammation. In this work we tested the action of H2S donor (GYY4137) on the activation of human neutrophils by E. coli LPS. We estimated the changes in redox status (ROS level, intracellularglutathione, NO), apoptosis and mitochondrial potential of neutrophils under the LPS action in the presence and absence of GYY4137. GYY4137 reduces the ROS level, slightly reduces GSH, does not influence the NO level and has no apoptogenic effect. LPS induce the increasing of ROS level and inhibit spontaneous apoptosis of neutrophils. We found that GYY4137 prevents the growth of ROS caused by LPS and leads to a reduction of LPS-induced inhibition of neutrophil apoptosis. Thus the mechanism of GYY4137 protection against inflammation, triggered by bacterial infection, is concerned with the neutralization of LPS effect on neutrophils.

Vascular grafts made of polytetrafluoroethylene and polyethylene terephthalate have widely been used in cardiovascular surgery. The causes of delayed colonization of such grafts by endotheliocytes and mesenchymal stem cells have not been adequately investigated. The authors examined the effect of polyethylene terephthalate on the functional activity of human bone marrow mesenchymal stem cells and endothelial progenitor cells in vitro. Proliferation (MTT assay, real-time cellular impedance), migration (Boyden chamber, real-time cellular impedance), and nitric oxide production (spectrophotometciacally) by progenitor endothelial cells and mesenchymal stem cells were assessed with and without the presence of polyethylene terephthalate. The functional activity of the cells was shown to depend on the presence of polyethylene terephthalate in a well with cells. Thus, polyethylene terephthalate turned out to exhibit a toxic effect on progenitor endothelial and mesenchymal stem cells. Treatment of grafts with gelatine or fibronectin improved colonization of grafts with cells.

The development of type 2 diabetes mellitus (DM2) is accompanied by disturbances in lipid metabolism. These include the increase in serum levels of atherogenic fractions of very low-density (VLDL) and low-density lipoproteins (LDL), total cholesterol, triglycerides and apo B. In contrast, the level of antiatherogenic high density lipoproteins (HDL) and the content of apolipoprotein A-I (apoA-I) decreased. To study the effect of the observed metabolic changes on insulin secretion in vitro, we used the islets of Langerhans isolated from the rat pancreas. It has been found that incubation of the islets in the presence of serum of the obese patients and patients with decompensated DM2 leads to a decrease in insulin secretion by 2.4 and 5.0 times, respectively. On the contrary, the addition of HDL to the incubation medium increased the insulin secretion by 3.4 times. A similar effect was observed in the presence of apoA-I, the main protein component of HDL. In the presence of apoA-I, the extracellular activity of matrix metalloproteinases (MMPs) demonstrated a 10-fold increase. The addition of LDL and VLDL to the islets did not change the secretion of insulin and activity of MMP. Our results testify to the important role of HDL and apoA-I in regulation of the insulin secretion by b-cells and the activity of MMPs in the islets of Langerhans.

The effects of chorionic gonadotropin (hCG) on the expression of the hTERT gene in combination with the conversion of the phenotype of naive T-cells and T-cells of immune memory in vitro were studied. hCG inhibited expression of hTERT mRNA in naive T-cells (CD45RA+) and immune memory T cells (CD45RO+), causing a decrease in the replicative potential of the cells. The presence of hCG in the culture led to the conversion of the phenotype of T-lymphocytes. hCG reduced the number of proliferating T-cells of immune memory, estimated by phenotypic signs by differential gating. hCG (10 IU/ml and 100 IU/ml) inhibited expression of CD25 by the studied populations, but did not modulate expression of the CD71 proliferation marker. Thus, hCG inhibited the functional activity of naive T-cells and T-cells of immune memory, which, in the context of pregnancy, can contribute to the formation of immune tolerance to the semi-allogenic fetus.

The use of sulfated polysaccharides from brown seaweed Fucus evanescens as adjuvants (native fucoidan in combination with polyphenols, fucoidan without polyphenols, a product of enzymatic hydrolysis of fucoidan) stimulated the formation of specific antibodies to the surface antigen of the hepatitis B virus (HBs-AG). Immunization of mice with vaccine compositions containing HBs-AG and fucoidan samples resulted in increasing the serum level of the pro-inflammatory (TNF-a, IFN-g, IL-2) cytokines. Increased production of these cytokines was detected in the culture of splenocytes additionally stimulated in vitro by fucoidans or phytohemagglutinin. The adjuvant effect of fucoidan and its structural modifications was comparable to that of the traditional licensed adjuvant aluminum hydroxide. The obtained results indicate a promising use of sulfated polysaccharides from F. evanescens as vaccine adjuvants.

Activity of telomerase catalytic subunit hTERT (human Telomerase Reverse Transcriptase) can be regulated by alternative splicing of its mRNA. At present time exact mechanism of hTERT splicing is not fully understood. Apoptotic endonuclease EndoG is known to participate this process. EndoG expression is induced by DNA damages. The aim of this work was to investigate the ability of DNA-damaging agents with different mechanism of action to induce EndoG expression and inhibit telomerase activity due to the activation of hTERT alternative splicing in normal activated human CD4+ and CD8+ T-lymphocytes. All investigated DNA-damaging agents were able to induce EndoG expression. Cisplatin, a therapeutic compound, producing DNA cross-links induced the highest level of DNA damages and EndoG expression. Incubation of CD4+ and CD8+ T-cells with cisplatin caused the changes in proportion of hTERT splice variants and inhibition of telomerase activity.

Muscle extracts of some fish species, i.e. pike (Esox lucius), sterlet (Acipenser ruthenus), pink salmon (Oncorhynchus gorbuscha) and, to a lesser extent, perch (Perca fluviatilis) and Russian sturgeon, (Acipenser gueldenstaedtii) prevent the development of premature senescence of the human embryonic fibroblasts induced by the sublethal concentration of H2O2. Muscle extracts of other fish species tested, i.e. coho salmon (Oncorhynchus kisutch) and zander (Sander lucioperca), have not demonstrated this feature. Cell proliferation increased after the action of the senescence-inhibiting muscle extracts. Possible mechanisms of the action of nature biologically active compounds that interfere with the development of stress-induced cell senescence are discussed.

In murine bone-marrow stromal microenvironment cells and in human multipotent mesenchymal stromal cells (MMSCs), proinflammatory cytokine interleukin-1 beta (IL-1β) serves as a growth factor. In murine bone tissue, IL-1β expression increases in vivo after irradiation. Here, we have presented our evaluation of the effects of exogenous IL-1β on the expression of NF-kB transcription factors in human MMSCs and stromal layer cells of murine long-term bone marrow cultures (LTBMCs). The cytokine signaling pathway was also activated in murine LTBMC by braking electron radiation in doses of 3-12 Gy. The level of expression of genes that code for IL-1β, IL-1β type-I receptor and NF-kB and IKK protein families have been studied at different time points post exposure. In both human and murine stromal cells, exogenous IL-1β led to an increase in the level of expression of its own gene, while levels of expression of NF-kB and IKK gene families were not substantially changed. Nevertheless, in human cells, a significant correlation between levels of expression of IL-1β and all NF-kB family genes was detected. It points to a similarity in IL-1β signal pathways in mesenchymal and hematopoietic cells, where the posttranslational modifications of NF-kB transcription factors play a major role. The irradiation of murine LTBMC resulted in a transient increase in the expression of genes that code NF-kB transcription factors and IL-1β. These results indicate an important role of Rel, Rela, Relb, and Nfkb2 genes in the induction of IL-1β signal pathway in murine stromal cells. An increase in IL-1β expression after the irradiation of stromal cells may be related to both the induction of inflammation due to massive cell death and to a profound stimulation of the expression of this proinflammatory cytokine expression.

The level of DNA single strand breaks, chromosomal abnormalities and sister chromatid exchanges and the possibility of its normalization with oligopeptide bioregulator Livagen and cobalt ions in the lymphocyte culture from patients with breast cancer have been studied. The results show that the genome of ductal breast cancer patients is characterized by the high density of DNA single strand breaks, high frequency of chromosomal abnormalities and increased levels of chromatin condensation. The usage of Livagen and cobalt in the form of modifying agents has a protective effect by all studied parameters. The obtained results allow us to conclude that research of lymphocytes of ductal breast cancer patients using the analysis conducted by us, can be useful in assessing the therapeutic effect in the treatment of breast cancer patients.

Aim: To study a neuroprotective effect of mexidol on the cell model of glutamate stress., Material and Methods: Cytological studies of an effect of glutamate stress on cerebellar granule cells were carried out., Results: Mexidol increased neuronal survival after the addition of glutamate by 8-10% (p<0.05). The effect of mexidol was more pronounced at the stage of neuron culture growth (5 days), cell survivability increased on average by 20%., Conclusion: The results of the study confirmed the neuroprotective effect of mexidol in the neuronal culture in glutamate toxicity model.

Cultivation of cells under artificial conditions is one of the necessary procedures when working with stem cells. At this stage, the cells lose control by the macroorganism, becoming independent systems. Therefore, during the passaging of the cells, the probability of undesirable modifications is increased. At present, there is not enough data on the first indications about modification of the cultured cells and when they can appear. In the work, the rabbit medullary mesenchymal stromal cells (MSCs) were studied during 5 passages after producing primary culture. The area and the spreading coefficient of a cell were used to evaluate the cell state. The measurements were made 1 h and 1 day after cell reseeding by analyzing digital images of intact living cells. During this time interval, the absolute values of the cell area increased with the passage number, whereas the increment of the cell growth area did not depend on the passage number. It is possible to distinguish three groups of cells. Smaller cells (the 1st group) prevail in cell population of the 1st passage. Next, their proportion is decreased and the proportion of larger cells is increased. Passaging of the cells does not influence their spreading coefficient.

Aim: to investigate the role of heterogeneous fibroblasts in the development of epiretinal membrane in eyes with modeled proliferative vitreoretinopathy., Material and Methods: The material for investigation were 6 eyes of 3 Chinchilla rabbits. Suspended fibroblasts (fibroblasts of the human skin - 200000 cells in 0.1 ml) were injected into the vitreous cavity via the pars plana. The animals were followed up for 1 month and then made out of the experiment. The eyes were enucleated and fixed in 10% neutral buffered formalin for routine histological examination. Microscopy was performed on the Leica system., Results: The main clinical and morphological criteria for a rabbit model of PVR induced by intravitreal injection of heterogenic fibroblasts have been established: epiretinal membrane formation, changes in intraocular structures (the retinal pigment epithelium and retina), and inflammation (due to transplantation immunity). Particularities of the epiretinal membrane development and the role of different intraocular structures have been described., Conclusion: The experimental fibroblastic model of PVR reproduces the final, fibrous, stage of PVR, which is significant for efficacy evaluation of antiproliferative drugs.

The study included apparently healthy people, conventionally healthy people predisposed to atherosclerosis, and as well as people with preclinical atherosclerosis (50 subjects in each group). Monocytes were isolated from whole blood and transferred to culture followed by studying pro- and anti-inflammatory activation of monocytes in response to stimulation by interferon-gamma and interleukin-4, respectively. As a marker of pro-inflammatory activation was the level of secretion of tumour necrosis factor-α (TNF-α) in the culture medium measured by means of immunoenzymatic assay. Chemokine CCL18 served as a marker of anti-inflammatory activation. We also examined secretion of a series of other chemokines and cytokines: MCO-1, IL-6, IL-1β, IL-8, GM-CFS, and others. Pronounced individual differences of cytokines and chemokines secretion were revealed in all groups. We carried out assessment the degree of altering secretion of cytokines and chemokines by stimulated monocytes compared with unstimulated culture. This approach may serve as an effective tool for assessment an individual reaction of congenital immunity.

The goal of our work was to examine content of sirtuin 1 in human skin at different ages to uncover a role of sirtuin 1 in aging of human skin. Sirtuin 1, proliferating cells nuclear antigen (PCNA) were detected by indirect immunohistochemistry in sections of the skin of human fetuses died antenatally from 22 to 40 weeks of pregnancy and humans from birth to 85 years old, died from various causes. Our results showed that the level of sirtuin 1 in dermal fibroblasts was decreased from prenatal period to 85 years old. Both the number of fibroblasts and their proliferative activity were also decreased through life. Age-related decrease in sirtuin 1 content in dermal fibroblasts is statistically significant correlated with age-dependent decrease in proliferation. Therefore, lowering of sirtuin 1 content in dermal fibroblasts occurring with age can be regarded as a mechanism which leads to inhibition of proliferation of dermal fibroblasts.

Skin aging is one of the topical issues in modern gerontocosmetology. Application of cosmetic products with short peptides is a promising measure for retardation of skin aging. This research is aimed at investigation of KE (Lys-Glu, Vilon) dipeptide influence on the expression of markers of aging in human skin fibroblasts in vitro. Collagen type I and sirtuin-6 expression in «young» and «old» skin cell fibroblasts cultures was studied using immunofluorescence confocal microscopy method. The areas of expression of collagen type I and sirtuin-6 are known to decrease in skin fibroblasts with aging by 3,5 and 3,6 times accordingly. KE dipeptide increases collagen type I expression area in «old» skin fibroblasts cultures by 83%. KE dipeptide increases expression area of sirtuin-6 in «young» and «old» skin fibroblasts cultures by 1,6 and 2,6 times correspondingly. Thus, KE dipeptide promotes functional activity of skin fibroblasts and inhibits their aging.

Aim: to study the effects of urokinase plasminogen activator (UPA) on the human retinal pigment epithelium (hRPE) cell culture., Material and Methods: The toxicity of 50 U/ml UPA was studied with the trypan blue exclusion test. Cell migration was assessed by the wound healing and modified Boyden chamber assays. Additionally, cell morphology, trypsin resistance, and Ki67 expression were investigated., Results: Trypan exclusion test did not reveal any cytotoxicity of 50 U/ml UPA against hRPE cells. The agent appeared able to induce cellular cluster formation and increase the number of spindle-shaped cells (6.4±2.4 cells/field and 67.3±3.2 cells/field in the controls and in the presence of 50 U/ml UPA, respectively, p<0.001). Cell migration in the Boyden chamber also showed a statistically significant increase (1.75-fold, p=0.012). Monolayer wounds were found to heal at an accelerated rate (p<0.05). This effect was dose-dependent, just like the increase in Ki67-positive cells (from 2.5 to 50 U/ml). Moreover, there was a reduction in trypsin resistance of the hRPE cells (the number of resistant cells in the control and 50 U/ml UPA cultures was 5.2±1.7 cells/field and 0.46±0.32 cells/field, respectively, p<0.001)., Conclusion: UPA, at concentrations of 50 U/ml or less, demonstrates no cytotoxicity against the hRPE cells. The effects of UPA on hRPE include stimulation of epithelial-mesenchymal transition, migration, proliferation, and intercellular interaction. At that, changes in migratory and proliferative activity are dose-dependent.

The liver has a marked capacity for regeneration. In most cases the liver regeneration is determined by hepatocytes. The regenerative capacity of hepatocytes is significantly reduced in acute or chronic damage. In particular, repair mechanisms are not activated in patients with alcoholic cirrhosis. Organ transplantation or advanced methods of regenerative medicine can help such patients. The promising results were obtained in clinical trials involving patients with various forms of liver disease who received transplantation of autologous bone marrow stem cells. However, to improve the effectiveness of such treatment it is necessary to search for more optimal sources of progenitor cells, as well as to evaluate the possibility of using descendants of these cells differentiated in vitro. In this study we isolated stromal cells from the liver biopsies of three patients with alcoholic cirrhosis, conducted their morphological and phenotypic analysis, and evaluated the hepatic potential of these cells in vitro. The stromal cells isolated from fetal liver were used for comparison. The results of this can serve as a basis for the development of a new method for the treatment of end-stage liver disease. The stromal cells isolated from the liver biopsies for a long time proliferate in a culture and this which makes it possible to expand them to large amounts for subsequent differentiation into hepatocyte-like cells and autologous transplantation.

Barriergenesis is the process of maturation of the primary vascular network of the brain responsible for the establishment of the blood-brain barrier. It represents a combination of factors that, on the one hand, contribute to the process of migration and tubulogenesis of endothelial cells (angiogenesis), on the other hand, contribute to the formation of new connections between endothelial cells and other elements of the neurovascular unit. Astrocytes play a key role in barriergenesis, however, mechanisms of their action are still poorly examined. We have studied the effects of HIF-1 modulators acting on the cells of non-endothelial origin (neurons and astrocytes) on the development of the blood-brain barrier in vitro. Application of FM19G11 regulating expression of HIF-1 activity and GSI-1 suppressing gamma-secretase and/or proteasomal activity resulted in the elevated expression of thrombospondins and matrix metalloproteinases in the developing blood-brain barrier. However, it caused the opposite effect on VEGF expression thus promoting barrier maturation in vitro.

Currently there is no single answer on the question of the participation of nitric oxide and NO-synthase (NOS) in the cardioprotection in the early phase of ischemic preconditioning (IP). In the present review the results of experimental studies indicate the involvement of NOS in the protective effect of IP and the literature data refuting this assumption. In addition, it was presented data on the increase of nitrates and nitrites (the stable metabolite of NO) in myocardial tissue after the IP, an increase in the expression of genes encoding NOS immediately after the IP, the data on the elimination of the protective effect of IP of NOS blockers and of imitation of IP affects IP by nitric oxide donors. It was exhibited a brief analysis of some of the studied mechanisms involving NO and NOS in cardioprotection at IP.

It is known that in various cell types bacterial lipopolysaccharide (LPS) causes mitochondrial disfunction and promotes accumulation of triglycerides in intracellular lipid droplets. The precise mechanisms which mediate LPS-induced neutral lipid deposition remain poorly understood. In the present work performed on primary cultured epithelial cells isolated from the frog urinary bladder we studied the possible role of mitochondrial reactive oxygen species (mROS) in LPS-in-duced alteration of lipid metabolism. It was shown that LPS stimulated ROS production, decrea- 705 sed fatty acids oxidation, enhanced intracellular triglyceride deposition and promoted the formation of lipid droplets visualized by Nile red staining. Pretreatment of cells with mitochondrial-tar-geted antioxidant MitoQ at dose 25 nM for 2 h almost completely eliminated all the above effects of LPS. In contrast to MitoQ, a-tocopherol was ineffective. The data obtained indicate that increase of mROS level is a critical factor that mediates LPS-induced intracellular deposition of neutral lipids in epithelial cells.

The influence of Nothrombel on the expression of membrane complex GPIb-IX-V of platelets activated by thrombin was studied. It is established that Nothrombel reduced thrombin-induced expression of complex GPIb-IX-V on the membrane of the activated platelets. The inhibitory activity of Nothrombel on the expression of GPIb (CD42b) was comparable to its activity with respect to GPIX (CD42a) and is equal to 43 %. With regard to GPV (CD42d) inhibitory effect of the drug was less pronounced and is equal to 22 %. The possible targets for Nothrombel are activation of thromboxane signaling pathway, as well as GPIb-IX-V receptors of platelets itself.

Unlabelled: Low-intensity laser radiation can be used as one of the methods for the non-specific regulation of the human mesenchymal stem cell (MSC) activity at the preliminary stage of their in vitro cultivation. The objective of the present study was to estimate the influence of the limiting regimes of continuous low-intensity laser radiation (CLIR) of red (635 nm) and green (525 nm) spectra., Material and Methods: The adhesive culture of human mesenchymal stem cells obtained from a donor's umbilical cord tissue was used in the experiments (following 4 passages). They were irradiated using a Lazmik-VLOK laser therapeutic device equipped with the KLO-635-40 (635 nm, 4,9 mW/cm(2)) and KLO-525-50 (525 nm, 5,4 mW/cm(2)) laser diode emitting heads operating in a continuous mode. A special nozzle (jar) for laser and vacuum massage (KB-5, 35 cm in diameter) was employed to fix the heads. The exposure time in all the irradiation regimes was 5 minutes., Conclusion: The study has demonstrated that neither the morphological features nor the viability of mesenchymal stem cells was altered under the influence of laser irradiation at the aforementioned energy and time parameters. The data obtained indicate that laser irradiation with the limiting levels of the chosen energy parameters produces no positive effect on the cell proliferative activity; more than that, it may cause its inhibition.

Previously, it was found that blood plasma extracellular DNA (ecDNA) of patients with rheumatoid arthritis (RA) is enriched with CpG-rich genomic DNA fragments, which contain TLR9 ligands (Veiko et al., 2006). In this study we have demonstrated that ecDNA of a RA patient and model fragments added to a cultivation medium of peripheral blood mononuclear cells (PBMC) of healthy donors stimulate expression of genes for the TLR9-MyD88-NF-kB signaling pathway; this leads to a significant increase in concentrations of the proinflammatory cytokines IL-6 and TNF-a in the cultivation medium. Human genomic DNA non-enriched with the CpG sequences did not stimulate IL-6 and TNF-a synthesis in PBMC. A scheme explaining the potential role ecDNA in the induction and maintenance of increased levels of the proinflammatory cytokines under conditions damaging the human cells has been proposed.

Specifics and key regulation mechanisms of compartmentalised protein synthesis in dendrites are reviewed. The up-to-date literature data of the subject are analysed. The results of many molecular, cytological and physiological experiments are presented. Also there is some information about a number of neurological diseases connected with dendritic translation regulation malfunction.

The dose-dependent effects of cytokines (IL-2, IL-7, and IL-15), which have a common g-chain, on mRNA expression of U2afll4 and GFi1 genes involved in regulation of alternative splicing of the Ptprc gene, have been investigated in vitro using T-lymphocyte cultures with different degrees of differentiation. IL-2, IL-7, and IL-15 caused a similar unidirectional inhibitory effect of various severity on restimulated CD45RO+ T-cells exposed to an antigen-independent activation; they caused a dose-dependent decrease of the U2af1l4 gene expression, and an increase of Gfi1 gene expression. This may suggest formation of active forms of the CD45 receptor, and also limitation of the formation of low-molecular short splice variants of the CD45RO receptor. Under conditions of antigen-independent stimulation of naive CD45RA+-cells rIL-7 and IL-15 exhibited opposite effects on U2af1l4 and Gfi1 gene expression. The increase of IL-7 concentrations in the incubation medium of naive cells was accompanied by a decrease in expression of both genes. IL-15 IL-7 exhibited opposite effects. Cytokines possessing a common g-chain (IL-2, IL-7, and IL-15) prevented antigen-independent differentiation of naive T-cells, by preventing the formation of polyclonal "surrogate" cells. In general, the study of the molecular mechanisms of genetic control determining homeostatic processes of T-cells in response to exposure to antigenic or non-antigenic treatments may be important for construction of a general model of self-maintenance and differentiation of immune cells.

Methemoglobin formation was examined in erytrocytes of 16 patients with Parkinson`s disease (PD) (stage 3-4 by the Hoehn and Yahr scale). The patients receiving levodopa-containing drugs (madopar, nakom) were also treated with intramuscular injections of mexidol (daily dose 100 mg/day) for 14 days. Control group included 12 clinically healthy persons. The erythrocyte methemoglobin content was determined by electronic paramagnetic resonance (EPR) using the EPR signal intensity with g-factor 6.0. The methemoglobin content was significantly higher in erythrocytes of PD patients than in healthy donors. The complex therapy with mexidol normalized the methemoglobin content in erythrocytes of PD patients. Incubation in vitro of erythrocytes of donors and PD patients with acrolein increased the methemoglobin content, while incubation with carnosine normalized the methemoglobin content in erythrocytes of PD patients. Prophylactic (i.e. before acrolein addition) and therapeutic administration of carnosine to the incubation system with acrolein decreased the methemoglobin content to its initial level. Results of this study suggest that inclusion of the antioxidants mexidol and carnosine in the scheme of basic therapy of PD may reduce side effects associated with methemoglobinemia.

The effect of 10- and 60-min cold stress was analyzed on reactive oxygen species (ROS) formation, peritoneal macrophage production of cytokines and corticosterone level under the opiate receptor blockade. It was observed that mice exposed to -20 °C for 10 min demonstrated the ROS inhibition, the effect was leveled against the background of naloxone introduction. The animal group subjected to 60-min stress on the contrary, showed the naloxone-dependent promotion of ROS product. Both variants of cold stress did not render significant effect on IL-1ß production. Against 60-min cold stress the TNF-α production was naloxone-dependently stimulated both in spontaneous and zymosan-induced cultures. IL-10 production by macrophages was improved irrespective of the cold stress duration, as well as stimulation presence or absence; and was abolished against a background of opiate receptor blockade. Therefore, the directionality of acute cold stress on the macrophage functions was dependent both on the parameters'' analyzed and on the duration of the exposure while the opiate receptor blockade significantly modified the immunoregulatory effects of hypothermia.

Development of hyaluronic acid (HA) supplemented biomaterials for regenerative medicine is complicated with easy water solubility of HA. It results in decreased stability of HA-based scaffolds that, consequently, initiated search for crosslinking techniques intended to retain HA within a scaffold. In this study, gels composed of type I collagen and native high molecular weight HA were prepared and their stability in buffer solutions has been evaluated. Fibronectin and human dermal fibroblasts were incorporated into gels as biological linkages for HA instead of chemical agents. Gel stability was estimated based on HA diffusion toward a buffer solution. In addition, HA and HA-supplemented gels resistance to fetal calf serum (FSC) as a part of cultural media for fibroblasts has been studied and no degradation of HA by serum hyaluronidase has been detected under the experimental conditions. The results showed that HA diffuses from collagen gels within 1—3 days. Neither fibronectin nor fibroblasts added separately prevent HA diffusion. But if gels were inoculated with fibroblasts together with fibronectin, more HA was left in the gels according to alcian blue staining. Agarose gel electrophoresis has shown that conditioned media collected from such gels are enriched with HA that differs from the initial one in that it contains a new portion of HA with higher molecular weight. It is concluded that HA-supplemented collagen gels are not stable in buffer solutions but fibroblasts incorporated into such gels synthesize significant amount of HA under the influence of fibronectin and partly compensate the loss of HA due to diffusion.

Metabolic activity of cells within a neurovascular unit is among the factors determining structural and functional integritY of the blood-brain barrier and the an- giogenesis process. in order to verify the hypothesis about the role Of g1YcolYtic activity in the perivascula astroglialcells associated with lactate release in the development of functioning of cerebral microvessel endothelial cells, we have used a three-component model of the brain neurovascular unit in vitro. The cells o f n o n -en d o th elia l o rig in w ere in c u b a te d in th e p rese n ce o f m o d u la to rs o f la c ta te pro d u c n ago ni glu c ose ta a G ly c o s o) , bas t h e oe t a n t a at- blocker of monocarboxylate transporters MCTlprCT and recepltiors of3Ctate0produasan (2-donisyoflactate G e8 breceptor) Iasa estbishe vthat that te suppression of lactate production and transport, prdc o1,adrcpin(C-O-Aa n (2gdoxysgflucoase as a glycolysis inhibitor), transport (phloretin as a sukr of lacaroduto transport , aswellasastimultionof3lactate receptors in astroglial cells, lead to aberrant development of endothelial layer, ther by u g g e tin t h efor atio o f anti ngi gencmi roen ircm ent for cerebral endothelium due to inappropriate lactate-m ediated effects. KeYw.ords:-n-eur-ovascular unit; metabolism; glYcolysis; lactate.

Changes in amplitudes of evoked inhibitory postsynaptic currents (eIPSCs) from rat cultured hippocampal neurons were studied using whole-cell patch-clamp technique in postsynaptic neuron and local extracellular electrical paired pulse stimulation of single presynaptic axon. Paired pulse depression (PPD) and paired pulse facilitation (PPF) were observed in studied 43 neurons. According to coefficient of variance (CV) analysis was found that CV of second respond was significantly larger than CV first by 57 % (n 26) during depression and significanty smaller by 27 % (n = 17) during facilitation. Thus, only presynaptic mechanism underlies short-term depression and facilitation. We also estimated quantal parameters assuming that transmitter release is reasonably described by a binomial distribution. We found that under our experimental conditions there were no significant changes in GABA release probability during PPD and PPF. The values of mean quantal content and mean number of sites of transmitter release were the same for first stimulus in pair and were equal 9.7 in depression and 5 in facilitation of synaptic transmission. The values both of them for second stimulus were significant decreased by 28 and 26% during PPD and were significant increased by 14 and 11% during PPF, respectively. Thus, there is significant difference in quantal parameters characterizing GABA release during short-term depression and facilitation of synaptic transmission.

The purpose of the work was to assess the influence of the concentrate of red table wine Saperavi on the cultivated nerve cells exposed to glutamate. The selection of Saperavi as the source of phenolic compounds was not accidental: this type of grapes in the Krasnodar region has the highest content of them - up to 4-5 g/dm3 and more. Polyphenol concentrate was prepared by pre-distillation of ethanol using vacuum, evaporation of red table wine with a rotary vaporizer, with the subsequent lyophilic drying. By the methods of voltammetry and chemiluminescence an antioxidant activity, and a quantity of antioxidants in the concentrate have been determined. By HPLC it was established that a large group of phenolic compounds with high antioxidant activity was present in the concentrate of polyphenols: procyanidins (total concentration up to 425 mg/dm3), quercetin (21.8-32.6 mg/dm3), gallic acid (124.2-164.7 mg/dm3), resveratrol (6.26-13.22 mg/dm3), catechins (1026 - 1480 mg/dm3). Effect of red wine Saperavi concentrate on glutamate cytotoxicity was studied in the neuron culture of the cerebellum of 7-9-day-old rats. It was shown that the presence of. antioxidants reduced the intensity of chemiluminescence in model systems that generate free radicals. It was established that quenching of chemiluminescence in the system <> composed 68.43%, and in the system of the yolk lipoproteins - 86.36%. The application of concentrate Saperavi significantly increased the survivability of neurons: at the doses 5, 10 and 30 mcg/ml the number of intact neurons was respectively 38.6; 41.5 and 37.1%. The dose 20 mcg/ml was the most effective - the proportion of live neurons comprised 47.4%. The obtained results can be explained by the high antioxidant activity of concentrate flavonoids, including high content of biologically active compounds - catechins, quercetin, rutin, resveratrol. Thus, the consumption of red wine in quantities that exclude harmful effects, can have a positive impact on human health and the brain in particular.

Registration of the changes in the functional metabolic status of neutrophils under the influence of low-intensity laser radiation (LILR) can be useful for the choice of the exposure parameters in the studies designed to evaluate immunotropic effects. Such investigations, in their turn, are very promising in clinical terms since their results can be used to optimize the laser therapy techniques. The objective of the present study was to evaluate the degranulation potential of neutrophils and the intensity of their NADPH-oxidase reaction in response to the exposure to laser radiation as exemplified by the release of lysosomal granules in vitro. The source of radiation was the LASMIK device operated in the continuous mode at the following parameters: wavelength 635 nm, power density 0.12 mW/cm2, exposure time 10, 30, 90, 120, 150 s in the case of the automatic timer control and 100 s in the case of manual shutdown. It was shown that the maximum lysosomal activity and release of lysosomal granules took place at a 90-10 s exposure and a wavelength of 635 nm with the appearance of a plateau within 120 and 150 s after the onset of irradiation. In the case of a shorter exposure of neutrophil granulocytes to LILR (10 and 30 s) no pronounced effect was observed. It means that low-intensity laser radiation of the red spectrum with a wavelength of 635 nm is a physical stimulus reinforcing exocytosis of lysosomal granules by neutrophils in vitro. The reliable changes of NADPH-oxidase activity of neutrophil granulocytes isolated from donor peripheral blood are recorded at an optimum exposure time of 90-100 s. It is concluded that the laser therapy techniques, at least those designed to regulate neutrophils should be applied with the optimum exposure time of 90-100 s.

It was investigated the effect of two adamantane derivatives, memantine and 5-hydroxyadamantan-2-one (5-HA), in patients with cerebrovascular disorders. In vitro studies showed that 5-HA, unlike memantine, exhibited antiplatelet activity. Experiments showed that memantine reduced cerebral blood flow in the brain cortex of intact rats and those under conditions of transient global ischemia, whereas 5-HA only selectively improved blood flow in ischemic brain and was superior to the reference drug nimodipine. The obtained data indicate the leading role of the GABA-ergic (rather than glutamatergic mechanisms) in implementation of the anti-ischemic cerebrovascular activity.