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Aim. This study aimed to examine the effect of Larifan on metabolic characteristics of human blood monocytes and granulocytes in vitro. Methods. Four healthy adult men aged 21-26 years were recruited to participate in the study as blood donors. The metabolic profile of human blood monocytes and granulocytes was evaluated by phagocytic activity, reactive oxygen species production, nitric oxide generation, and arginase activity. Phagocytosis of FITC-labeled inactivated Staphylococcus aureus and reactive oxygen species generation were estimated by flow cytometry. Arginase activity was assessed in cell lysates, and nitric oxide generation in supernatants was examined using the Griess reaction. Results. Phagocytic index and reactive oxygen species generation were found to be lower in both human blood monocytes and granulocytes treated with Larifan. The drug caused a dose-dependent increase in nitric oxide production, as well as a decrease in the arginase activity of blood monocytes. Conclusions. Our results indicate the ability of Larifan to reinforce the antiviral properties of resting phagocytes along with containment of oxidative stress development.

Members of Ralstonia solanacearum species complex (RSSC) are causal agents of vascular wilt disease in more than 450 crop species, including solanaceous plants such as potatoes, tomatoes, bell pepper, eggplant, etc. These phytopathogens cause serious yield loss mostly in solanaceous crops which are grown in tropical, subtropical, and temperate regions of the world. Yield losses comprise 80%–100% in potato, up to 91% for tomato, 10%–30% in tobacco, 33%–90% in banana, and reduce crop productivity and yield. PCR-methods are specific, sensitive and cost-effective approaches for the detection and identification of RSSC members. The objective of this study was to compare specificity of routinely used primer mix for PCR RSSC detection with the newly developed pairs of species-specific primers for ease of use diagnostics in a laboratory. Materials and Methods. The conserved genomic regions of the 16S rRNA sequences of R. solanacearum, R. pseudosolanacearum, and R. syzygii were selected for the design of primers for this study. Newly created primer species specificity was tested in PCR using the DNA of the two targets and 13 non-target strains of bacteria. Results. Three pairs of newly created primers Rs-28(F)/Rs-193(R), Rs-28(F)/OLI-160(R), Rs28(F)/OLI248(R) produced single specific fragments for bacterial strains of Ralstonia solanacearum: 166 bp, 132 bp, and 220 bp. products respectively. No PCR products were obtained during amplification with the negative control or non-target DNA templates from other bacterial species. Conclusion. Designed primers can be used for the development of PCR system for the qualitative and quantitative detection of RSSC members.

Hematological and immunological markers of systemic inflammation were studied in 6-hydroxydopamine (6-OHDA)- and lipopolysaccharide (LPS)-induced models of Parkinson’s disease (PD). Experiments were carried out on adult male Wistar rats: 1 – intact animals; 2 – sham-operated animals and 3 – 6-OHDA- and LPS-lesioned animals. PD development was confirmed by the results of behavioral testing (apomorphine test, open field test) and immunohistochemical detection of the loss of dopaminergic neurons. Hematological indices (complete blood count and differential leukocyte count (DLC)) were examined using hematological analyser. Immunological indices included phenotypic (CD206 and CD80/86) and metabolic (oxidative metabolism and phagocytic activity) characteristics of circulating monocytes (Mo) and granulocytes (Gr), which were determined by flow cytometry, as well as plasma levels of C-reactive protein, which were determined by ELISA. LPS-induced PD was associated with neutrophilia, 1.9 times increased neutrophil-to-lymphocyte ratio, 3 times increased platelet-to-lymphocyte ratio, and 3 times increased systemic immune inflammation index as compared to intact animals. Functional profile of circulating phagocytes from LPS-lesioned animals was characterized by the pro-inflammtory metabolic shift, as was indicated by 5 times increased oxidative metabolism indices and up-regulated CD80/86 expression along with decreased phagocytic activity and CD206 expression. 6-OHDA-lesioned rats demonstrated decreased DLC indices as compared to intact and sham-operated rats. Functional profile of circulating phagocytes in this model was characterized by anti-inflammatory shift. The results obtained from this study demonstrated that stereotaxic LPS-induced PD is appropriate rodent model for the study of systemic inflammation which is inherent for the disease pathophysiology.

Aim. This study aimed to investigate the circadian rhythm of the murine peritoneal macrophage (PM) metabolic profile. Methods. The metabolic profile of PM was characterized by phagocytic activity, reactive oxygen species (ROS) generation, and by the expression of phenotypic markers, associated with a pro- and anti-inflammatory metabolic shift. Phagocytosis of FITC-labeled inactivated Staphylococcus aureus, ROS generation, CD80, CD86, and CD206 expression were estimated by flow cytometry at a regular 4h interval over the daily light-dark cycle. Results. The phagocytic index and percentage of ROS-producing PM were found to be lower in the resting phase (ZT4) as compared to the active phase. In contrast, the level of CD86 expression was the highest in the inactive phase (ZT8). There was also a statistically significant peak in the proportion of ROS-producing PM, as well as in the level of ROS production per cell at the time of awakening (ZT12). As opposed to ROS generation, ZT12 was characterized by the lowest level of cell-surface CD206 expression. Conclusions. Our results indicate that there is a circadian rhythm in functional polarization of murine PM with an anti-inflammatory activation state in the resting phase in comparison to the active phase.

Cancer metastasis and recurrence are the leading causes of cancer-related death. Tumor cells which leave the primary or secondary tumors and shed into the bloodstream are called circulating tumor cells (CTC). These cells are the key drivers of cancer dissemination to surrounding tissues and to distant organs. The use of CTC in clinical practice necessitates the deep insight into their biology, as well as into their role in cancer evasion of immune surveillance, tumor resistance to chemo- radio- and immunotherapies and metastatic dormancy. Aim. The purpose of the work was to review the current knowledge on the CTC biology, as well as the prospects for their use for the diagnosis and targeted treatment of metastatic disease. Methods. The work proposed the integrative literature review using MEDLINE, Biological Abstracts and EMBASE databases. Results. This review summarizes and discusses historical milestones and current data concerning СTС biology, the main stages of their life cycle, their role in metastatic cascade, clinical prospects for their use as markers for the diagnosis and prognostication of the disease course, as well as targets for cancer treatment. Conclusions. Significant progress in the area of CTC biology and their use in cancer theranostics convincingly proved the attractiveness of these cells as targets for cancer prognosis and therapy. The effective use of liquid biopsy with quantitative and phenotypic characteristics of CTCs is impeded by the imperfection of the methodology for taking biological material and by the lack of reliable markers for assessing the metastatic potential of CTCs of various origins. The variety of mechanisms of tumor cells migration and invasion requires the development of complex therapeutic approaches for anti-metastatic therapy targeting CTCs. Efforts to address these key issues could help developing new and effective cancer treatment strategies.

Deep insight into compositional and functional features of endophytic bacterial communities residing in wheat grains opens the way to the use of their plant growth promoting and biocontrol abilities in agricultural biotechnology. The aim of this work was to compare grain-residing endophytes from winter wheat varieties with different sensitivity to Pseudomonas syringae pv. atrofaciens (McCulloch) and to examine their plant-beneficial traits and antagonistic effects. Grain-residing bacteria were isolated from surface-sterilized grains of three wheat varieties sown in Ukraine following a culture-dependent protocol, and were screened for their plant growth promotion (PGP) and antagonistic properties. Bacterial morphotypes were represented by gram-negative rods, endospore-forming bacilli and gram-positive cocci. Different resistance to phytopathogenic pseudomonads was associated with distinctive quantitative and functional features of grain-residing endophytic communities. High resistance to P. syringae was coupled with the prevalence of gram-negative rods in the endophytic community, the highest proportion of endophytic bacteria possessing three PGP activities (phosphate solubilization, nitrogen fixation and production of indolic compounds) simultaneously, and with the most potent antagonistic activity of grain-residing endospore-forming bacilli. In total, five grain-residing isolates, which were obtained from three wheat varieties (two isolates from varieties with medium and high resistance and one – from a low-resistant variety), demonstrated ability to restrain P. syringae pv. atrofaciens (McCulloch) growth. Two isolates (P6 and P10) which were obtained from the high-resistant wheat variety Podolyanka and were assigned to Paenibacillus and Brevibacillus genera according to their biochemical profiling and MS-DS identification, showed the most potent antagonistic effects as indicated by maximum inhibition zone in agar well diffusion assay. These results shed light on the association of the features of grain-residing endophytic bacteria with wheat resistance to phytopathogenic pseudomonads. Isolates from the high-resistant wheat variety can be recommended for grain dressing as plant growth promoting and biocontrol agents for P. syringae pv. atrofaciens (McCulloch).

The aim of the work was to explore fatty acid composition of purslane seed water extract and its effect on the metabolic profile of murine peritoneal macrophages. Fatty acid composition was evaluated by gas chromatography–mass spectrometry. Collection of murine macrophages from the peritoneal cavity was done without preliminary sensitization. Reactive oxygen species generation was assayed by flow cytometry and nitroblue tetrazolium test. Phagocytic activity was evaluated by flow cytometry. Nitric oxide production was analyzed in cell supernatants by Griess reaction. Arginase activity was measured in cell lysates by standard colorimetric assay. Reactive oxygen species and nitric oxide production were significantly lower in murine macrophages simultaneously treated with purslane seed water extract and lipopolysaccharide in comparison to macrophages treated with lipopolysaccharide only. Also, the studied extract caused statistically significant increase in arginase activity of unsensitized peritoneal macrophages. That is consistent with the fatty acid content of this extract, since it contained comparatively higher proportion of unsaturated fatty acids exhibiting anti-inflammatory properties, than saturated fatty acids known for their pro-inflammatory effects.

Herbal preparations are widely used in the complementary treatment of inflammatory diseases including hepatobiliary disorders, among which chronic cholecystitis is one of the most common. This study was aimed to investigate the effect of the water extract from polyherbal composition consisting of eight medicinal plants (Helichrysum arenarium, Mentha piperita, Calendula officinalis, Taraxacum officinale, Polygonum aviculare, Matricaria chamomilla, Portulaca oleracea, Hypericum perforatum) on biochemical and haematologic indices as well as on immune reactivity parameters in patients with chronic cholecystitis. It was found that consumption of polyherbal tea during 45 days was followed by normalization of liver biochemical indices (serum level of cholesterol, alanine aminotransferase and aspartate aminotransferase activity, total and unconjugated bilirubin levels and diminution of hematological (leukocytosis, increased erythrocyte sedimentation rate and neutrophil-to-lymphocyte ratio) and immunological (increased serum level of pro-inflammatory cytokines) signs of systemic inflammation in patients with hepatobiliary disorder. The results of the research showed that the studied polygerbal composition can be recommended as a component of the complementary therapy for patients with chronic cholecystitis.

Hepatobiliary system diseases represent an important medical and social problem due to increasing morbidity rates worldwide. Liver and biliary diseases are characterized by complex pathophysiology as well as by multi- and comorbidity. The treatment of such diseases necessitates multitarget drug development. The effectiveness of current drugs in the treatment of hepatobiliary disorders remains low and the incidence of side-effects are profound. This actualizes the search and development of highly effective hepatoprotectors with a low incidence of side effects. Medicinal plants potentially constitute a sourse of such preparations. The review summarizes the data concerning mechanisms of hepatoprotective and immunomodulatory effects of medicinal plants and their phytoconstituents. The prospects for the development and use of herbal remedies in the treatment of hepatobiliary diseases are outlined.

The aim of the work was to explore phytochemical characteristics of water extract from polyherbal composition based on P. oleracea and it’s effect on oxidative metabolism of murine peritoneal macrophages. The qualitative phytochemical analysis was conducted by colorimetric method. Quantitative analysis of phenols was performed in the test with gallic acid as a standard. Murine peritoneal macrophages were isolated without previous sensitization. Leukotoxicity of the water extract from polyherbal composition leukotoxicity was evaluated in MTT test. Reactive oxygen species generation was assayed by the nitroblue tetrazolium reduction method. Phytochemical analysis revealed the presence of water-soluble and insoluble phenols, tannins, saponins, flavonoids, cardiac glycosides and coumarins in the studied plant mixture. The water extract from polyherbal composition used in a range of concentration 1–1000 μg/ml (lyophilisate in distilled H2O) didn’t exhibit any toxic effects on murine peritoneal macrophages. Water extract from polyherbal composition caused statistically significant dose-dependent increase in oxidative metabolism of murine peritoneal suggest modulatory effect of studied water extract from polyherbal composition on innate immunity cells.

Glioma-associated microglia/macrophages (GAM) represent an attractive therapeutic target for the development of the alternative methodology in the treatment of gliomas. This study was aimed to investigate the effect of intranasally administered TLR3 agonist Larifan on microglial cell metabolic profile in rats with C6 glioma. Our results demonstrate progressive generation microglial cell population with immunosuppressive and pro-inflammatory properties in C6 glioma-bearing brain. Intranasally delivered TLR3 agonist is capable to abrogate the creation of this pro-tumoral immune infiltrates, probably, through the effect on myeloid-derived suppressor cells, and can be considered as a promising agent for glioma therapy aimed the GAM re-education.

The aim of the work was to investigate the effect of the intracranial catheter placement on the metabolic profile of rat microglia. Microglial cells were isolated by the centrifugation in Percoll gradient. Oxidative metabolism and phagocytic activity were investigated by flow cytometry. Arginase activity was examined by colorimetric method. Nitrite level was assayed in Griess reaction. It was found that intracranial catheter placement caused down-regulation of nitrite synthesis by 3 times, augmentation of the reactive oxygen species generation by 1.5 times, and slightly decreased microglia phagocytic activity. Thus, intracranial catheter placement causes long term anti-inflammatory shift of microglia metabolism in rats.