Engineering of Glyceraldehyde-3-Phosphate Dehydrogenase with Dual NAD/NADP Cofactor Specificity for Improving Amino Acid Production.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme in the central metabolism of microbial cells. GAPDHs differ in cofactor specificity and use NAD, NADP, or both cofactors, reducing them to NADH and NADPH, respectively. Sufficient NADPH supply is one of the critical factors required for synthesis of the amino acids l-lysine, l-threonine, and l-proline in industrially important -based producer strains.

[The development and application of reagents kit for detection RNA of the causative agents of melioidosis and glanders by transcription-based amplification (NASBA) in real-time.].

The polymorphism of clinical manifestations of melioidosis and glanders and their high mortality require improvement of diagnostics for detection of this agents. The perspectivity of development of transcription-based amplification real-time NASBA diagnostic kits is determined by high analytical sensitivity and the opportunity to perfom the verification of the results of other methods for pathogenic Burkholderia species detection. The fragment of 23S rRNA gene was selected as the target for development of real-time NASBA kit.

[The comparative evalution of informativeness of immunologic and molecular genetic methods and means during stages of specific indication of melioidosis agent].

The reference-center of monitoring of agents of glanders and melioidosis carried out testing of reagents kits for diagnostic of agent of melioidosis and other close-related species of Burkholderiae in vitro. At the stage of specific identification of pathogenic Burkholderiae the diagnostic possibilities of commercial and experimental kits of reagents for express- and rapid analysis were evaluated. The criteria of evaluation of diagnostic value of kits of reagents were sensitivity, specificity and time of implementation of studies.

[Immunogenic properties of experimental production series of Staphylovac-2 vaccine].

Aim: Study experimental production series of Staphylovac-2 by accumulation of specific IgG and safety.

Materials And Methods: Experimental production samples of staphylococci vaccines were studied by the accumulation of specific IgG in sera of immunized BALB/c line mice in EIA. Safety was evaluated in tests of acute and chronic toxicity including pathomorphologic and histologic, hematologic and biochemical studies, studies of the effect on central nervous system.

[Immunobiological properties of Bordetella pertussis lipopolysaccharide in the acellular pertussis vaccine].

Aim: Study of Bordetella pertussis lipopolysaccharide (LPS) immunobiological properties in the acellular pertussis vaccine.

Materials And Methods: Experimental series of acellular pertussis vaccines (APV), lyophilized LPS were used. Antibody titers against LPS in mice sera were evaluated by using EIA with peroxidase conjugate of anti-species antibodies against mice IgG.

[Preclinical studies of an adsorbed diphtheria-tetanus-pertussis vaccine (ADTP-vaccine) with acellular pertussis component].

Aim: Evaluate standardness of antigenic composition of pertussis component, completeness of sorption of pertussis, diphtheria and tetanus components, specific activity and safety of experimental series ofADTP-vaccine with acellular pertussis component (ADTaP-vaccine).

Materials And Methods: The content of separate antigens (pertussis toxin, filamentous hemagglutinin and agglutinogens 1, 2, 3) in samples of acellular pertussis component of ADTaP-vaccine and completeness of sorption of pertussis component of ADTaP-vaccine were evaluated by using enzyme immunoassay. Completeness of sorption of diphtheria and tetanus components were determined in flocculation reaction and antitoxin-binding reactions, respectively.

[Experimental study on chemotherapy of acute glanders].

Glanders is a zoonotic infection inducing acute forms of the disease (pneumonia, sepsis) in humans and animals under certain conditions, which even with the use of modern chemotherapy have unfavourable prognosis. Insufficient of efficacy of antibiotics with in vitro low MIC for planktonic bacterial suspension of Burkholderia mallei in chemotherapy of acute forms of glanders was due to the capacity of the pathogen for intracellular survival and formation of biofilms. Under such conditions the susceptibility of B.

[The use of polymerase chain reaction for detection of the agents of glanders and melioidosis using experimental infection].

Glanders and melioidosis are severe infectious diseases of people and animals. The causative agents of these infections refer to the potential agents of bioterrorism of group B. In this work the possibility of use of flagellin-based primers for the identification of B.

[Use of infusoria Paramecium caudatum for the evaluation of the toxicity of antigens of different causative quarantine agents].

The results of the evaluation of the toxicity of bacterial antigens obtained from the causative agents of plaque, glanders, melioidosis, cholera on infusoria of the species P. caudatum, as well as on cell lines L-929, CHO K-1 and peritoneal macrophages of BALB/c mice, are presented. As revealed in this study, the method of toxicity determination on infusoria is similar in its sensitivity to the methods of testing on.

[Ultrastructural immunochemical study of pathogenic Burkholderia capsule].

The capsular structures of Burkholderia pseudomallei, B. mallei, B. cepacia and their avirulent noncapsular mutants were studied with the use of electron ahd immunocytochemical techniques.

[Experimental study on the possibility of using live tularemia vaccine to increase resistance to heterologous infection disease].

In experiments on guinea pigs immunized with Francisella tularensis 15, or live tularemia vaccine (LTV), the level of heterologous protective effect to dangerous infectious diseases caused by Yersinia pestis, Burkholderia pseudomallei, B. mallei, Mycobacterium tuberculosis was studied. The study revealed that during the first 4 weeks after the subcutaneous immunization with LTV the level of resistance of the immunized animals to heterologous infective agent reliably increased as indicated by the survival rate of the animals, as well as by the survival time of those killed by infection, in comparison with the controls.

[Dependence of mode of DNA binding to benzo crown derivatives of actinocin on the size and distance from the chromophore of crown fragments].

Complexes of DNA with benzocrown derivatives of actinocin were studied by viscometry and dynamic birefringence. Changes in the macromolecular structure of DNA caused by complex formation were determined. Models of DNA binding to the studied compounds were suggested on the basis of data obtained.

[Burkholderia thailandensis: biological properties, identification and taxonomy].

Burkholderia pseudomallei-like microorganisms have been isolated from soil and water in regions with endemic melioidosis. These strains have biochemical and antigenic profiles identical to melioidosis agents, except that they differ by virulence and L-arabinose (vir-, ara+). There are minor differences between these species by rRNA sequence.

[Immunotropic and immunogenic properties of Burkholderia pseudomallei surface and membrane antigens].

The immunotropic and immunogenic properties of some chromatographic fractions of B. pseudomallei surface antigenic complex, as well as the preparations of B. pseudomallei outer and cytoplasmic membranes, were studied.

[The design of a diagnostic test system based on magnetic sorbents and liposomes].

The results obtained in the study of the possibility of using magnetic sorbents for the construction of a diagnostic assay system based on the antigen-antibody interaction are presented. As a model, Yersinia pestis capsular antigen and immunoglobulins to it have been used. A solid-phase immunofluorescent liposomal assay method has been developed; this method can be used for the detection of biopolymers in the sample under study and for the determination of their activity.

[Study of protective properties of antigen-containing liposomes of different lipid composition in plague].

Immunization of animals with a liposomal capsular and major somatic antigen of a plague causative agent enhanced their protective effects against plague infection. Lipopolysaccharide incorporation into the lipid membrane resulted in an increased protective effect of antigen-containing liposomes, which may be related to the single delivery and interaction with immune-competent cells. Incorporation of phospholipids with high phase transfer temperature and positively charged stearylamine by the membrane facilitates the immunogenicity of agents.

[The action of gamma irradiation on trypsin immobilized on a modified polypropylene textile material].

In studying the proteolytic activity and ESR spectra of gamma-irradiated samples of trypsin immobilized at an inoculated copolymer of polypropylene with polyacrylic acid it was established that the carrier of a modified polypropylene increases the radioresistance of trypsin immobilized on it.

[Entrapping of hydrophobized plague capsular antigen into large unilamellar liposomes].

The entrapping of a plague capsular antigen into the liposomes, prepared by the "reverse phase" evaporating method, was investigated. Both the native antigen and the antigen modified by palmitoylchloride were used. The entrapping of the modified antigen into the liposomes was greater than that of the native one: 46.

[Various properties of urease encapsulated into liposomes].

The enzymic activity of plant urease encapsulated into liposomes from egg lecithin was studied. Liposomes contained 3-5% of the initial enzymic preparation. Incorporation of urease into liposomes increases the permeability of the lecithin membrane for urea.

[Recanalization of the hepatic ducts in high obstructive blockage].

Forty three patients were subjected to 48 operations for restoration of the passability of lobular ducts when the latter were obturated by tumor (36 patients) and by alveococcus (7 patients). Ten patients died immediately after operation. Death was caused by acute hepato-renal insufficiency and acute pancreatitis.