Preparative Production and Purification of Recombinant Human Cyclophilin A.

In this work, we developed the method of preparative production of recombinant human cyclophilin A (rhCypA) in Escherichia coli. The full-length cDNA encoding the gene of human CypA (CYPA) was amplified by RT-PCR from the total RNA of human T cell lymphoma Jurkat. The nucleotide sequence of CYPA was optimized to provide highly effective translation in E.

Development of an immunoassay test system based on monoclonal antybodies and immunomagnetic particles for the detection of F. tularensis cells.

Tularemia is an especially dangerous infection caused by the gram-negative bacterium Francisella tularensis. It belongs to natural focal infections, and therefore is under continuous control by quarantine services. When carrying out their activities they use a whole range of diagnostic tools.

[The becoming of reactions of lipolysis in phylogenesis. The palmitic and oleic triglycerides as substrates. Insulin, condition of normolipemia and formation of hyper lipoproteinemia type IIb, IV and V].

According to phylogenetic theory of general pathology, when living in ocean all were carnivorous (piscivorous) fatty acids transferring to cells in form of non-polar triglycerides nitially began apoB-48 chylomicrons, continued lipoproteins of very low and low density and fnalized its apoB-100 endocytosis. The fatty acids are transferred by chylomicrons + lipoproteins of very low density + lipoproteins of low density and non-polar triglycerides are hydrolyzed by hepatic glycerolhydrogenase and co-enzyme apoC-III; according WHO classifcation, hyperlipoproteinemia corresponds to type V. On land, in herbivorous who are not yet synthesized insulin, apoB-48 and chylomicrons left process of non-polar triglycerides transferring.

[The individual fatty acids of blood plasma: biological role of substrates, parameters of quantity and quality, diagnostic of atherosclerosis and atheromotosis.].

The atherosclerosis and atheromotosis are supposed to be, according to phylogenetic theory of general pathology, two etiologically different aphysiological processes, unified by community of pathogenesis. The atherosclerosis is a derangement of biological function of trophology (feeding), biological reaction of exotrophy (external feeding) and biological function of adaptation, biological reaction of compensation in response to deficiency of ῳ-3 and ῳ-6 polyenoic fatty acids. In case of deficiency of polyenoic fatty acids in cells and during synthesis of eicosanoids of group I from unsaturated endogenous ῳ-6 С20: 3 digomo-γ-linoleic unsaturated fatty acid, atherosclerosis is developed, a complex metabolism disorder in vivo.

[The clinical trials of agar and Mossel EE broth.].

The cultural diagnostic of acute intestinal infections is based on application of growth medium for selective cumulation of enterobacteria. The development of composition and technology of production of national import-substituting growth medium is an important task for supporting laboratories with high quality medical products. In the state research center of applied microbiology and biotechnology are developed and registered in established procedure a dry growth medium for selective cumulation of enterobacteria (Mossel EE broth) and dry growth medium for selective isolation and counting of enterobacteria (Mossel agar).

[The perspective fluoroplastic porous carrier for conservation and transfer of samples of blood plasma: application of "dry drop" technique with the purpose of detection of individual fatty acids.].

It is proposed for preserving samples of biological fluids (blood plasma first of all) and mailing them to analytical laboratory to significantly modify a well-known technique of "dried blood spot" (DBS) at the expense of application of home-made fluoroplastic filtering material type MFFK-G as a porous carrier of fluid biological sample. The synthetic material contains no chemically active molecular fragments and it has four times greater water capacity than filter and chromatographic paper. Thus, fluoroplastic porous disc of mass of 15 mg is suitable to carrying of 70-100 mkl (instead of 15-20 mkl) of fluid.

Effect of Audiogenic Seizures on the Dynamics of Fatty Acid Composition of Hippocampal Dental Gyrus in Krushinsky-Molodkina Rats.

Changes in the fatty acid composition of the hippocampal dentate gyrus in Krushinsky-Molodkina rats with hereditary predisposition to audiogenic seizures were studied in 1, 3, and 14 days after 1 or 5 seizures. Seizure activity changed the content of saturated and monounsaturated as well as polyunsaturated fatty acids at different terms after seizures. After seizures, the content of individual fatty acids changed in different directions.

[The positional specificity of individual triglycerides-substrates for lipase action. The oleic triglycerides of palm oil, palmitic triglycerides of milk, ions of calcium and magnesium.].

The counter-insulin effect of surplus of palmitic fatty acid in food is implemented under: a) formation in vivo of palmitic type of fatty acids metabolism with deficiency of substrate for ATP synthesis and permanent shortage of energy for accomplishment of biologic functions; b) compensatory activation of biologic function of adaptation, biologic reaction of compensation. The activation with catecholamines in visceral fatty cells of gland the hormone-dependent lipase which is not blocking insulin, increases content of unesterified fatty acids in blood plasma. Until in blood plasma the level of unesterified fatty acids is increased the cells phylogenetically justified stop absorption of glucose along with development of hyperglycemia and hypertriglyceridemia - insulin resistance syndrome.

[The positional isomers of palmitic and oleic triglycerides, lipolysis, conformation of apoB-100, formation of apoE/B-100 ligand and absorption of lipoproteins of very low density by insulin-dependent cells under action of statins.].

The study was carried out to trace back quantitative alterations of positional isomers, oleic and palmitic triglycerides and individual fatty acids in blood serum. After two weeks of taking of Simvastatin (40, 80 mg), analysis of blood serum established decreasing of content of Рhosphatidylcholine and more reliable decreasing of amount of non-etherized alcohol cholesterol. No alterations of content of particular fatty acids were detected.

[The insulin initiates "kinetic perfection" of biologic function of locomotion. The glucose as substratum for synthesis of ω-9 oleic fatty acid by cross-striated miocytes.].

The phylogenesis theory affords ground for the following propositions. 1. There is no absorption of glucose from intercellular medium by cells in vivo until there is possibility to absorb polar fatty acids from associates with albumin.

Adsorption chromatography of proteins.

A method for adsorption chromatography of proteins is proposed. A protein solution is passed through a cellulose column at a pH value corresponding to an isoelectric point of the protein. Depending on the charge of unwanted proteins, they either remain at the origin (if charges of protein and ion-exchanger are opposite) or are released from the column (if charges of protein and ion-exchanger coincide).

Ionogenic groups in the active site of lysostaphin. Kinetic and thermodynamic data compared with x-ray crystallographic data.

The active site of lysostaphin is shown to contain a residue of glutamic acid. As judged by a pK value of 9.2 (with pentaglycine bridges in peptidoglycan of staphylococci as a substrate), another ionogenic residue could be the epsilon-amino group of a lysine.

Cloning and expression of protective antigens of Mycobacterium tuberculosis Ag85B and ESAT-6 in Francisella tularensis 15/10.

The possibility of expression of genes encoding mycobacterial antigens in Francisella tularensis 15/10 vaccine strain cells has been shown for the first time. To obtain stable and effective expression of mycobacterial antigens in the F. tularensis cells, the plasmid vector pPMC1 and hybrid genes consisting of the leader part FL of the F.

Preventive and therapeutic effects of alpha-acid glycoprotein in mice infected with B. anthracis.

We studied the effects of alpha1-acid glycoprotein preparations on the survival rate of BALB/c mice infected with the lethal dose of B. anthracis STI-1. Apart from native alpha1-acid glycoprotein from donor blood, we studied 3 glycoforms differing in the affinity for concanavalin A and structure of carbohydrate chains.

Uncommon properties of pLD105 conjugative plasmid.

Plasmid pLD105 isolated from a clinical strain of E. coli determines nitrofuran resistance due to inactivation of low-molecular-weight nitrofuran reductase subunit. pLD105 plasmid belongs to IncF.

Michaelis-menten kinetics for determining enzymatic activity of lysostaphin.

The rate of lysostaphin-catalyzed lysis of staphylococci follows the Michaelis-Menten equation at [E](0) << [S](0), i.e., the activity of the enzyme is proportional to its concentration.

[Kinetic analysis of behavior of chemotactic bacterial population at the interface of two media].

The behavior of a population of bacteria at the boundary with a chemotactically active water medium containing the sites of specific binding to cell receptors was analyzed. A kinetic model of chemotaxis was used for the analysis. Differences in the behavior of strains metabolizing and not metabolizing the substrate were revealed.

[Factors of Yersinia pestis providing circulation and persistence of plague pathogen in ecosystems of natural foci. Communication 2].

To maintain continuous circulation of plague pathogen in natural foci, the pathogen should be capable of invading host organism, resisting the bactericide protective systems of rodent, and reproducing itself to maintain the content of bacteria at a level sufficient for further transmission by fleas to a new host. Each of these stages of the Yersinia pestis circulation is determined by a variety of factors of plague pathogen, which may act either individually or in combination. Each of the factors itself may be involved in the pathological process at different stages of its development or in pathogen transmission.

[Genetic typing of Mycobacterium tuberculosis strains by spoligotyping and genome fingerprinting techniques].

A total of 122 M. tuberculosis clinical drug-resistant strains isolated in Central Russia were studied by spoligotyping and genome fingerprinting techniques. According to spoligotyping results 77% of M.

[Biological properties of clinical isolates of Mycobacterium tuberculosis].

The invasiveness of 2 grud sensitive and 8 holy-resistant M. tuberculosis clinical isolated was evaluated in experiments on BALB/c mice. Mycobacterial suspension was injected into the caudal vein of experimental animals.

Structure of Cry3A delta-endotoxin within phospholipid membranes.

Interaction of delta-endotoxin and its proteolytic fragments with phospholipid vesicles was studied using electron microscopy, scanning microcalorimetry, and limited proteolysis. It was shown that native protein destroys liposomes. The removal of 4 N-terminal alpha-helices and the extreme 56 C-terminal amino acid residues did not affect this ability.

[Immunomodulatory properties of 2-C-methyl-D-erythritol-2,4-cyclopyrophosphate and search for its new derivatives].

A strong immunomodulatory effect of 2-C-methyl-D-erythritol-2,4-cyclopyrophosphate (MEC) responsible for the survival of bacteria was shown on isolated macrophages and in experimental infections in mice (typhoid and tularemia). Derivatives of MEC were found by 1H-NMR spectroscopy under stress conditions in colorless mutants of the bacteria and isolated to be subsequently purified and used for modulation of the immune system of animals.

[Microfungicid--a preparation based on trichoderma viride for plant diseases control].

A technology was designed for manufacturing a preparation based on Trichoderma viride Pers ex S.F. Gray that strongly suppresses the development of causative agents of certain plant diseases and displays a growth-stimulating activity.

[Identification of nontuberculous mycobacteria by methods of molecular biology].

A number of clinical and laboratory strains of microorganisms belonging to different species of mycobacteria of the nontuberculous complex were tested with the use of the polymerase chain reaction and the restriction analysis. The unique restriction profiles of the following species of mycobacteria have been obtained: M. fortuitum VI, M.

[Effect of space flight conditions on properties of hydrocarbon-oxidizing bacteria].

Results of experiments on the Mir space station (EO-25 and EO-26) demonstrated that the conditions of orbital flight, primarily the cosmic radiation, was a mutagenic factor affecting both the genotype and phenotype of an oil-oxidizing bacterial strain, Mycobacterium flavescens EX-91. The emerging mutants differed from original culture by the rate of colony growth and the ability to ferment certain carbohydrates or synthesize beta-galactosidase. Changes in the rate of utilization of raw oil and individual hydrocarbon types (constituting model mixtures) suggest that cosmic radiation may serve as a means of obtaining mutant clones of microorganisms with new properties.