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Parkinson's disease (PD) is the second most common neurodegenerative disorder. Alpha-synuclein misfolding and aggregation resulting in neurototoxicity is a hallmark of PD. The prion properties of alpha-synuclein are still under discussion. Exosomes (extrcellular vesicles 40-100 nm in size) can play a key role in the transport of pathogenic forms of alpha-synuclein. The most frequent inherited forms of the disease are PD associated with mutation in the leucine-rich repeat kinase 2 (LRRK2-PD) and glucocerebrosidase (GBA-PD) genes. The aim of our work is to evaluate the concentration and size of exosomes derived from blood plasma of patients with GBA-PD, asymptomatic GBA mutation carriers, and the effect of GBA and LRRK2 mutations on alpha-synuclein level in exosomes derived from peripheral blood plasma. Plasma extracellular vesicles were isolated via chemical precipitation and sequential ultracentrifugation and characterized by transmission electron microscopy, nanoparticle tracking analysis (NTA), and flow cytometry. Total alpha-synuclein level in plasma exosomes was estimated by enzyme-linked immunosorbent assay. Patients with sporadic PD, PD with dementia, patients with inherited PD (GBA-PD, LRRK2-PD), and GBA mutation carriers were included in the study. The concentration on plasma exosomes was higher in GBA-PD patients that in sporadic PD patients, asymptomatic carriers of mutations on GBA gene, and control (p = 0.004, 0.019 and 0.0001 respectively). The size of plasma exosomes was higher in GBA-PD patients compared to asymptomatic carriers of GBA mutations and control (p = 0.009 and 0.0001, respectively). No significant difference was found for exosomal alpha-synuclein levels in the studied groups. Our results allowed us to suggest that a decrease in GBA activity may affect the pool of plasma exosomes, and mutations in the LRRK2 and GBA genes do not influence the level of plasma exosomal alpha-synuclein.

Objective – assessment of effectiveness of the measures for biological safety provision at potentially hazardous facilities in the territory of the Republic of Tatarstan in the process of preparation and holding of FIFA World Cup-2018 in Kazan. Materials and methods. Regulatory-legal acts covering the requirements to the provision of biological safety during the work with pathogenic biological agents, international guidelines and documents determining inter-agency collaboration between the Rospotrebnadzor Administration in the Republic of Tatarstan and other departments and inter-agency task forces (Office of the Federal Security Bureau of Russia in the Republic of Tatarstan, Counter-Terrorism Commission in the Republic of Tatarstan, etc) were studied. Results and discussion. Considered were managerial-methodological approaches and matters of inter-agency cooperation on biological safety provision during preparation and holding of FIFA World Cup-2018 in Kazan. Biological safety was examined through the prism of its provision while working with pathogenic biological agents at potentially hazardous biological facilities and counterterrorism integrity of the objects. The guiding, directive document in the performance of the complex of measures on biological safety provision and inter-agency collaboration was the Order of the President of the Russian Federation dated May 09, 2017 No 202 “On peculiarities of application of reinforced security measures during holding of FIFA World Cup-2018 and FIFA Confederations Cup-2017 in Russia”. As the result of the measures conducted, biological safety, including counter-terrorism security of potentially hazardous biological objects and sanitary-epidemiological welfare, was fully provided.

Parkinson's disease (PD) is the second most common neurodegenerative disorder. Alpha-synuclein misfolding and aggregation resulting in neurototoxicity is a hallmark of PD. The prion properties of alpha-synuclein are still under discussion. Exosomes (extrcellular vesicles 40-100 nm in size) can play a key role in the transport of pathogenic forms of alpha-synuclein. The most frequent inherited forms of the disease are PD associated with mutation in the leucine-rich repeat kinase 2 (LRRK2-PD) and glucocerebrosidase (GBA-PD) genes. The aim of our work is to evaluate the concentration and size of exosomes derived from blood plasma of patients with GBA-PD, asymptomatic GBA mutation carriers, and the effect of GBA and LRRK2 mutations on alpha-synuclein level in exosomes derived from peripheral blood plasma. Plasma extracellular vesicles were isolated via chemical precipitation and sequential ultracentrifugation and characterized by transmission electron microscopy, nanoparticle tracking analysis (NTA), and flow cytometry. Total alpha-synuclein level in plasma exosomes was estimated by enzyme-linked immunosorbent assay. Patients with sporadic PD, PD with dementia, patients with inherited PD (GBA-PD, LRRK2-PD), and GBA mutation carriers were included in the study. The concentration on plasma exosomes was higher in GBA-PD patients that in sporadic PD patients, asymptomatic carriers of mutations on GBA gene, and control (p = 0.004, 0.019 and 0.0001 respectively). The size of plasma exosomes was higher in GBA-PD patients compared to asymptomatic carriers of GBA mutations and control (p = 0.009 and 0.0001, respectively). No significant difference was found for exosomal alpha-synuclein levels in the studied groups. Our results allowed us to suggest that a decrease in GBA activity may affect the pool of plasma exosomes, and mutations in the LRRK2 and GBA genes do not influence the level of plasma exosomal alpha-synuclein.

At present, it is believed that disregulation of alpha-synuclein (SNCA) metabolism and its aggregation is associated with the pathogenesis of Parkinson’s disease (PD). It is thought that the death of dopaminergic neurons in PD can be mediated by the effect of dopamine on the process of alpha-synuclein oligomerization, primarily by direct oxidation of the protein. At the same time, the effect of dopamine on the regulation mechanism of the SNCA gene expression is not excluded. It is known that transcription factors GATA-1, GATA-2, and ZSCAN21 can participate in the regulation of SNCA expression. In the present study, we evaluated the effect of exogenous dopamine (100 μM) on the mRNA level of the SNCA, GATA-1, GATA-2, and ZSCAN21 genes and the level of total alpha-synuclein in cultured peripheral blood lymphocytes (PBL) of patients with PD (n = 18) and individuals of the control group (n = 14) using real-time PCR and ELISA, subsecuently. A decrease in the SNCA expression level under the action of dopamine was first found in PBL of patients with PD (P = 0.013), of the control group (P = 0.004), and of the combined group that included patients with PD and control individuals (P = 0.001). When assessing alpha-synuclein level, a tendency toward its decrease in PBL of individuals from the control group (P = 0.068), as well as of the combined group of PD patients and control individuals (P = 0.059), was revealed by comparing the cells treated and untreated with dopamine. An increase in the mRNA level of the ZSCAN21 gene in PBL of control group individuals (P = 0.022), as well as of the GATA-1 gene in PBL of the group of patients with PD (P = 0.019) cultured in the presence of dopamine, was shown. An increase in the mRNA levels of the genes GATA-1, GATA-2, and ZSCAN21 in PBL under the treatment with dopamine was also observed in the combined group of patients with PD and control individuals (P = 0.027, 0.029, and 0.002 for GATA-1, GATA-2, and ZSCAN21, respectively). Thus, the obtained data show the effect of dopamine on the mRNA level of the SNCA, GATA-1, GATA-2, and ZSCAN21 genes in cultured PBL of PD patients and control group individuals, suggesting a possible effect of dopamine on the regulation of SNCA expression.

Infectious agents collections used in practical work are to be securely protected as they can cause infections among people and animals; they are potential biological terrorism objects including periods when mass sport, political, or cultural events take place in the country. It made our research truly vital. Our research goal was to analyze an interdepartmental workgroup func-tioning aimed at inspecting objects located on areas where 2017 FIFA Confederations Cup and 2018 FIFA World Championship were to take place. This workgroup had to deal with biological substances, biological and microbiological organisms and infectious agents from II-IV pathogenicity groups. Its basic task was to minimize biological hazards during work with pathogenic biological agents. The research involved examination of legal documents (including international ones) which regulated biological safety requirements for work with pathogenic biological agents; consideration of joint work with the RF Federal Security Service, Tatarstan Regional Office, Tatarstan Anti-Terrorism Commission for providing anti-terroristic protection of organizations dealing with infectious agents, including those common for people and animals, on the example of Rospotrebnadzor Regional Office in Tatarstan; analysis of work performed by the interdepartmental workgroup. It is shown that efficient interdepartmental interaction allowed to provide biological safety in Kazan and on adjoining territories, to prevent pathogen entering the environment and possibility to use pathogenic biological agents as biological terrorism objects during preparation for 2017 FIFA Confederation Cup and the event itself.

Aim. To estimate the effect of transforming growth factor в, lipoprotein lipase, fibrinogen в and glycoprotein 3б gene polymorphisms on different variants of atherosclerosis severity. Methods. The study included 319 patients with angiografically verified atherosclerosis. In all patients FGB (rs1800788), LPL (rs328), GpIIIa (rs5918) and TGFВ (rs1800469) polymorphism genotyping was performed. In all patients we evaluated the influence of the studied gene polymorphisms on parameters of atherosclerosis severity such as occlusion and presence and severity of stenosis. Presence of occlusion, occlusion and/or two and more stenoses over 70%, occlusion and multiple critical stenoses were considered the severity criteria. Results. The studied gene polymorphisms were shown to be associated with severe course of atherosclerosis. GPIIIa polymorphism was commonly determined in patients with more severe course regardless of occlusion and number and severity of stenoses. FGB gene influenced the rate of occlusion development, TGF gene was expressed only in the group with occlusion and/or two and more stenoses over 70%, and LPL gene proved to be significant for the development of occlusion associated with critical stenosis. Conclusion. The frequency of rare alleles of polymorphic regions is significantly associated with different variants of severe atherosclerosis course; this could suggest unequal influence of gene polymorphisms as well as their proteins on different stages of atherogenesis and requires further investigation.

Aim. To study the influence of brain natriuretic peptide gene polymorphism (polymorphic locus Т-381С) on brain natriuretic peptide serum level and congestive heart failure onset risk and clinical features in patients with coronary heart disease. Methods. 412 patients with congestive heart failure were examined. Genotyping was performed by polymerase chain reaction. Brain natriuretic peptide N-terminal fragment level was assessed by ELISA. The control group included 211 healthy controls with no signs of cardiovascular pathology on examination. Results. In healthy people with C/C genotype the level of brain natriuretic peptide N-terminal fragment was significantly higher in comparison with people carrying T/T genotype. It was found that the T allele and T/T genotype of the T-381C natriuretic peptide gene polymorphic locus was associated with high risk, severity and unfavorable clinical course of congestive heart failure in patients with coronary heart disease. At the same time, the C allele and C/C genotype emerged as a protective factor regarding the risk, severity and clinical course of the disease. Conclusion. T/T genotype carriers of the of the T-381C natriuretic peptide gene polymorphic locus are a special subgroup associated with high risk of congestive heart failure onset and unfavorable clinical course. Therefore these patients with coronary heart disease should be considered as a group requiring an out-patient control and preventive measures targeted on congestive heart failure and premature mortality prevention.

The effect of cartolin (0.33 μM), an antistress regulator of cytokinin type, on the cytoskeleton-dependent changes in lectin activity in the roots of unhardened (23°C) and cold-hardened (3°C, 7 days) 7-day-old plants of three cultivars of winter wheat (Triticum aestivum L.) was studied. In unhardened plants, cartolin increased activity of soluble and cell wall-bound lectins in a cultivar-specific mode. This is evidently important for subsequent enhancement of adaptation processes in the cell. The inhibitor of microtubule polymerization, oryzalin, reduced the activity of soluble lectins and increased that of cell wall-bound lectins. A reduced sensitivity of lectin activity to oryzalin after cartolin treatment could result from its stabilizing action on the cytoskeletal structures and on the interaction between cell-wall lectins and microtubules. The most efficient cartolin action, the suppression of oryzalin effect on lectin activity in hardened plants, was observed in the frost-sensitive wheat cultivar. It is likely that cartolin treatment is more efficient in the activation of adaptation processes occurring with the involvement of cytoskeletal structures in the cultivars of lower tolerance.

The polypeptide composition and functional activity of cell-wall lectins from roots of winter wheat (Triticum aestivum L., cv. Mironovskaya 808) seedlings during cold hardening were studied. Several phases of lectin activity changes were observed, which indicates their involvement in the development of general adaptation syndrome of the cell. After 0.5-h low-temperature treatment, marked alterations occurred in the profile of protein elution: lectins with mol wts of 78 and 42.5 kD disappeared and new ones with mol wts of 72, 69, 37, and 34.5 kD appeared. It was established that 17.5-and 69-kD lectins and most lectins eluted with glucose were arabinogalactan proteins (AGP), which permitted a supposition that these lectins were involved in the interaction between the cell wall and cytoskeleton. After 7-day-long hardening, total protein content reduced and lectins with mol wts of 69 and 37 kD disappeared, which corresponded to reduced lectin activity by the end of hardening. A transient appearance of 37-and 69-kD lectins, which are AGP, might indicate their involvement in the triggering the development of plant-cell defense responses.

1-(2,3,5-Tri-O-acetyl)-beta-D-ribofuranosyl indole, the key compound in the synthesis of glycosides with the bis(indole) aglycone, was obtained for the first time by the indoline-indole method. There were synthesized 3-(1-methylindol-3-yl)-4-(1-glycosylindol-3-yl)furan(or pyrrole)-2,5-diones containing the residue of beta-D-ribofuranose or 2-deoxy-beta-D-ribofuranose and analogous glycosides of indolo[2,3-a]furano(or pyrrolo)[3,4-c]carbazol-5,7-diones, which are structurally relative to the antitumor antibiotic rebeccamycin. Their cytotoxicities toward a number of human tumor cell lines were studied in vitro, and the carbazole N-glycosides were shown to be more active than the bis-(indole) glycosides. At the same time, the ribofuranosides were found to be less active than the corresponding ribopyranosides synthesized previously.

The effect of oryzalin, a microtubule polymerization inhibitor (10 MM), on lectin and mitotic activities (mitotic index and duration of mitotic phases) was studied in unhardened (23 degrees C) and hardened (7 days, 2-3 degrees C) winter wheat seedlings. Three wheat cultivars differing in their frost tolerance were compared. Oryzalin treatment (3 h) decreased activity of soluble lectins, increased activity of cell wall lectin mitotic index. Under these conditions, prolongation of anaphases and disappearance of telophases were detected. Plant hardening reduced the sensitivity of cell wall lectins and mitotic activity to the cytoskeleton inhibitor due, presumably, to the appearance of cold-stable microtubules. Plant growing and hardening with oryzalin stopped mitoses and caused the appearance of polyploid cells and cells with micronuclei. These abnormalities were preserved after hardening. The results obtained demonstrate an important role of microtubules in adaptation of plants to low temperature.

Effects of oryzalin (10 microM), an inhibitor of microtubule polymerization, on the activity of soluble and cell wall lectins were studied in 7 day-old seedlings of unhardened (23 degrees C) and cold acclimated (7 days at 2-3 degrees C) winter wheat (Triticum aestivum L.). Seedlings were grown in the presence of 25 microM and 1 mM Ca2+, 500 microM verapamil, 250 microM chlorpromazine or without modifiers of calcium status in the medium. Inhibitor of the microtubule polymerization inhibitor, likely as inhibitors of Ca(2+)-signal, decreased the activity of soluble lectins and increased that of cell wall lectins. Apparently, injury of microtubule phosphorylation results in a more considerable microtubule disorganization, than that observed after oryzalin effect. A low Ca2+ concentration (25 microM) depressed, while a high concentration (1 mM) prompted microtubule sensibility to oryzalin. Such an effect of high Ca2+ concentration may be related to destabilizative action of Ca(2+)-calmodulin in these conditions, because chlorpromazine decreased oryzalin-induced increase in the activity of cell wall lectins with 1 mM Ca2+. It is concluded that the activity of cell wall lectins depends on the microtubule status that is regulated by calcium signal.

A synthesis of 3-(1H-3-indolyl)-4-(1-glycosyl-3-indolyl)furan-2,5-diones and -1H-pyrrole-2,5-diones modified with the residues of D-ribo-, D-xylo-, L-arabino-, D-galactopyranose, and D-lactose was described. Influence of the compounds prepared on DNA biosynthesis in CaOv cells was studied.

A synthesis of the derivatives of bis(indolyl)furan and bis(indolyl)pyrrole, which contain an acyclic fragment or a residue of D-ribo-, D-xylo-, L-arabino-, D-galactopyranose, or D-lactose was described. Bis(indole) aglycone was created using 1-glycosylindoles, which were synthesized by the indoline-indole method.

A number of compounds with valuable biological properties have been found among the pyrazolo[3,4-d]pyrimidines and their nucleosides. Thus, experimental antitumor activity and/or antiparasitic properties have been exhibited by nucleosides of 4-aminopyrazolo[3,4-d]. pyrimidine, allopurinol (4-oxypyrazolo[3,4-d]pyrimidine) [6], and 4-methylmercaptopyrazolo' [3,4-d]pyrimidine [3]. Of particular interest are the nucleosides of pyrazolo[3,4-d]pyrimidines which have residues of functional carboxylic acids in position 3 of the hetero ring. As has been illustrated by nucleosides of 4-substituted and certain 4,6-bisubstituted pyrazolo[3,4-d]pyrimidines, the introuction of a nitrile, carbamoyl, thiocarbamoyl and similar groups to position 3 results in greater cytotoxic and antitumor activity [2, 7, i0, ii].

A number of new 1-aryloxy-5-methyl-1,2,3,6-tetrahydro-1,2,6-phosphadiazine-1,3-diones were obtained from arylphosphoric acid dichlorides through the corresponding diamidophosphoric acid esters. Conversion of the 1-phenoxy and 1-ethoxy derivatives to 6-methyl-4-hydroxypyrimidine under Vilsmeierformylation conditions was observed. These compounds were thionated to give the corresponding 1,3-dithiones; the 1-phenoxy derivative was subsequently methylated in the 6 position and animated to give the phosphorus-containing analog of 6-methyl-2-thiocytosine. The bromination of the 1-ethoxy compound and replacement of the bromine by a secondary amine residue were studied.