SANCTIONS: POLITICAL AND ECONOMIC FORESHORTENING

A number of aspects of the change of the political and economic relations, apparent by the sanctions policy of the western states to the Russian Federation and its realization, has been considered. The balance between the liberty, equality and fraternity, the perfect competition and free business, on the one hand, and the competition of smothering, ball and chain, on the other hand, – has been disclosed. It has been substantiated, that the western states seek to substitute the colonial influence in the past for sanctions pressure in our days. It allows them to get not only the competitive advantage, but also to obtain the absolute dictatorship sometimes. The conclusion has been made, that external intervention in the natural course of managing and especially the rough administrative influence never gives a positive effect

Diversity and occurrence of methylotrophic yeasts used in genetic engineering

Methylotrophic yeasts have been used as the platform for expression of heterologous proteins since the  1980’s. They are highly productive and allow producing eukaryotic proteins with an acceptable glycosylation level.  The first Pichia pastoris-based system for expression of recombinant protein was developed on the basis of the treeexudate-derived strain obtained in the US southwest. Being distributed free of charge for scientific purposes, this system has become popular around the world. As methylotrophic yeasts were classified in accordance with biomolecular  markers, strains used for production of recombinant protein were reclassified as Komagataella phaffii. Although patent  legislation suggests free access to these yeasts, they have been distributed on a contract basis. Whereas their status  for commercial use is undetermined, the search for alternative stains for expression of recombinant protein continues.  Strains of other species of methylotrophic yeasts have been adapted, among which the genus Ogataearepresentatives prevail. Despite the phylogenetic gap between the genus Ogataeaand the genus Komagataellarepresentatives, it turned out possible to use classic vectors and promoters for expression of recombinant protein in all cases. There  exist expression systems based on other strains of the genus Komagataellaas well as the genus Candida. The potential  of these microorganisms for genetic engineering is far from exhausted. Both improvement of existing expression systems and development of new ones on the basis of strains obtained from nature are advantageous. Historically, strains  obtained on the southwest of the USA were used as expression systems up to 2009. Currently, expression systems  based on strains obtained in Thailand are gaining popularity. Since this group of microorganisms is widely represented  around the world both in nature and in urban environments, it may reasonably be expected that new expression systems for recombinant proteins based on strains obtained in other regions of the globe will appear

Development of Immune-Chromatographic Monoclonal Test-System for the Detection of <i>Yersinia pseudotuberculosis</i>, Serogroup I

Objective of the study was to develop monoclonal immunoassay for the detection of the pseudotuberculosis agent, serogroup I. Materials and methods. Specific components, that were used for immune-chromatographic test-system development were mouse monoclonal antibodies of hybrid cell lines, obtained to lipopolysaccharide antigen of the outer membrane of the pathogen’s «cold» variant (YP-101N2V4, YP-105S5A10); and rabbit anti-species antibodies against murine immunoglobulins. Particles, (30±2) nm in the diameter, were used to prepare colloidal gold-antibody conjugate. Antibody concentration for conjugation was 10-15 % greater than the D580 exit point on the plateau. For the production of immune-chromatographic test-system a set of membranes - MDI Easypack - manufactured by «Advanced Microdevice», India was deployed. Finished conjugate was applied onto the membrane by means of impregnation. Antibodies in the selected quantities were applied onto the analytical and control membranes via Dispensers. Substrates coated with the conjugate and ready-made working membranes were vacuum dried in a heat cabinet. Assembled immune-chromatographic test-systems were cut off 4.5 mm each and tested for specificity and sensitivity. Results and conclusions. Developed has been immune-chromatographic test-system for the detection of pseudotuberculosis pathogen, serogroup I. Utilized have been monoclonal antibodies of the hybrid cell line YP-105C5A10 in colloidal gold conjugate and monoclonal antibodies of the hybrid cell line YP-101H2B4 in the test line. The test-system allows for the detection of Y. pseudotuberculosis strains, serogroup I, at concentrations varying from 500 ths. m.c.·cm-3 (8 of the 11 strains under study) up to 4 million m.c.·cm-3 and does not identify closely related yersinia and heterologous microorganisms in quantities of 100 million m.c.·cm-3

Production of subtilisin proteases in bacteria and yeast

In this review, we discuss the progress in the study and modif ication of subtilisin proteases. Despite longstanding applications of microbial proteases and a large number of research papers, the search for new protease genes, the construction of producer strains, and the development of methods for their practical application are still relevant and important, judging by the number of citations of the research articles on proteases and their microbial producers. This enzyme class represents the largest share of the industrial production of proteins worldwide. This situation can explain the high level of interest in these enzymes and points to the high importance of designing domestic technologies for their manufacture. The review covers subtilisin classif ication, the history of their discovery, and subsequent research on the optimization of their properties. An overview of the classes of subtilisin proteases and related enzymes is provided too. There is a discussion about the problems with the search for (and selection of) subtilases from natural strains of various microorganisms, approaches to (and specifics of) their modif ication, as well as the relevant genetic engineering techniques. Details are provided on the methods for expression optimization of industrial subtilases of various strains: the details of the most important parameters of cultivation, i. e., composition of the media, culture duration, and the inf luence of temperature and pH. Also presented are the results of the latest studies on cultivation techniques: submerged and solid-state fermentation. From the literature data reviewed, we can conclude that native enzymes (i. e., those obtained from natural sources) currently hardly have any practical applications because of the decisive advantages of the enzymes modified by genetic engineering and having better properties: e. g., thermal stability, general resistance to detergents and specif ic resistance to various oxidants, high activity in various temperature ranges, independence from metal ions, and stability in the absence of calcium. The vast majority of subtilisin proteases are expressed in producer strains belonging to different species of the genus Bacillus. Meanwhile, there is an effort to adapt the expression of these enzymes to other microbes, in particular species of the yeast Pichia pastoris

Manufacturing of Hybridomas-Producers of Monoclonal Anti-Bodies to Tularemia Agent Antigens

Carried out have been two experimental studies on hybridization of mouse myeloma cells and lymphocytes of BALB/c mice immunized with inactivated microbe Francisella tularensis cultures. As a result obtained have been hybridomas-producers of monoclonal antibodies (MAb) specific to the antigens of tularemia agent. Evaluated have been the prospects of its application for the detection of the agent under discussion using enzyme-linked immunoassay. Established is the fact that monoclonal antibodies produced by 31G1F10, 32E5D3, 35B11C8, 36C2F11 hybridomas make it possible to identify microbe cells of various tularemia agent strains when concentrated up to 0.5·106 mc/sm3, and do not interact with cultures of heterologous microorganisms when concentrated to 1.0·108 mc/sm3, which testifies to their specificity. These MAb are planned to be used for the construction of immune-enzyme and immune-chromatographic test-systems designed for tularemia agent detection

Development of the Immuno-Enzyme Test-System for the Detection of <i>Legionella pheumophila</i>, Serogroup I

Developed is the highly sensitive and specific immuno-enzyme test-system, which is perspective for the detection of L. pneumophilia, serogroup 1. Isolated are the three hybrid cell lines that secrete monoclonal antibodies to specific epitopes of L. pneumophilia, serogroup 1 lipopolysaccharide antigen. Hyper immune rabbit sera, characterized by highly specific activity and specificity, are obtained using lipopolysaccharide antigen

Early predicting of preeclampsia in pregnant women after assisted reproductive technologies

Aim: to identify new prognostic criteria of potential preeclampsia (PE) in pregnant women after assisted reproductive technologies (ART) for timely PE prophylaxis.Materials and methods. A prospective study of 85 patients who entered the program of ART was conducted. All patients were examined for possible hemostatic abnormalities (genetic thrombophilia and chronic hypercoagulation) and also for granulocytemacrophage colony-stimulating factor (GM-CSF) in the serum during the most critical periods (4–6, 12–14, 22–24 и and 30–32 weeks) of the fetoplacental complex formation.Results. The lowest level of GM-CSF was observed in patients with hemostatic disorders. Thus, in pregnant women who later developed PE, there was a decrease in GM-CSF level below the physiological: in those diagnosed with genetic thrombophilia – by 79.4 %, and those with hypercoagulation – by 63.6 %.Conclusion. The determination of serum GM-CSF and identification of hemostatic abnormalities in pregnant women after ART has a prognostic importance for potential PE. This result is significant for the understanding of the pathogenesis of PE and also has a practical value: it allows the doctor to attribute the patient to a high risk group from the first trimester of her pregnancy and start preventive therapy rather early