New Advantageous Method for the Production of Purified Cholera Toxin B-Subunit and Monoclonal Antibodies to It

Put forward is an efficient method for manufacturing cholera toxin B-subunit. Its advantages are relative simplicity and economy feasibility, as well as maximum output of the purified B-subunit, absolutely free from toxic A-subunit contaminant. All this is due to the deployment of cholera vibrio recombinant strain producing only cholera toxin B-subunit instead of cholera toxin as it is, which results in lack of residual preparation toxicity. Applied has been gel-penetration column chromatography, providing for stable native state and maximum antigen output. The method under discussion is verified experimentally. Sample purity has been analyzed after each phase of chromatographic investigation on TSK gel HW-60, using disc electrophoresis. It is established that three steps of purification are ample for the obtainment of cholera toxin B-subunit preparation free from admixtures. Immunological activity of the purified B-subunit is validated by monoclonal antibody obtainment. Designed preparation of cholera toxin B-subunit and monoclonal antibodies to it can serve as a basis for the development of various immune-diagnostic test-systems alternatives

Characteristics of Phenotypic and Genetic Properties of <i>Francisella tularensis</i> 15 NIIEG Vaccine Strain with an Extended Storage Period

Investigated have been cultural-morphological, biochemical and genetic properties of lyophilized cultures of F. tularensis 15 NIIEG vaccine strain, accumulated within 60-years term and deposited at the State Collection of Pathogenic Microorganisms of Scientific Center on Expertise of Medical Application Products. The studies undertaken have demonstrated that storing of the strains in such a form at low temperatures, does not prevent changes of their genetic and phenotypic properties to the full extent. It is established that F. tularensis 15 NIIEG strain lyophilized in 1953, 1966, 1969, 2003 and 2012 maintains its immunogenic properties when cultivated on nutrient media Ft-agar with or without addition of blood, based on dissociation rates (87-99 %) of SR-colonies. While F. tularensis 15 NIIEG strain 1990 contains specified amounts (not less than 80 %) of immunogenic colonies if cultivated on nutrient media with the addition of blood, and fails to meet the requirements - if cultivated without. Identified in F. tularensis 15 NIIEG strain 1987 SR-colony decrement of 70-75 % in case of cultivation with or without addition of blood testifies to the deterioration of its immunogenic properties. RAPD and ERIC typing has showed high stability of the genome of F. tularensis 15 NIIEG cultures lyophilized at different times. Tularemia microbe vaccine strain has unique RAPD and ERIC profiles, insignificant alteration of which is observed upon storage of pathogen subculture in the dried from

Improvement of Approaches to the Verification of the Vaccine Strain <i>Francisella tularensis</i> 15 NIIEG during Long-Term Storage

The aim of the study was to improve the methods for verifying the vaccine strain Francisella tularensis 15 NIIEG during long-term storage under current conditions.Materials and methods. The paper summarizes the results of studying the phenotypic and genetic properties of lyophilized cultures of the vaccine strain F. tularensis 15 NIIEG (1953, 1966, 1969, 1987, 1990, 2003, 2012 and 2013) stored at SCEMAP for a period of one to 60 years.Results and discussion. Previous studies have revealed that freeze-dried cultures of F. tularensis 15 NIIEG generally had the characteristics of the vaccine strain, with the exception of deviations from the regulatory requirements for residual virulence and specific safety. The stability of preservation of deletions in the pilA and pilE genes (the region of differentiation RD19) and the genes encoding lpp lipoprotein (RD18) in the vaccine strain, which was stored for various periods of time in a lyophilized state, has been confirmed. The vaccine-strain-specific mutation C178404T (by the genome of F. tularensis LVS strain, GenBank NCBI no. CP009694) has been identified, and an approach to determine it has been developed. The data obtained are promising as regards using the above deletions in the RD18/RD19 regions in combination with the C178404T mutation to assess the authenticity of the vaccine strain using molecular genetic methods. Thus, the conducted retrospective analysis of the data on the cultures of tularemia microbe vaccine strain from the 1940s to 2013 and the gathered experimental data, made it possible to supplement the uniform requirements for the manufacture, study, maintenance, storage and movement of F. tularensis 15 NIIEG vaccine strain with new evidence. Based on the results obtained, the authors have drawn a draft methodological recommendations of the federal level “Vaccinal strain Francisella tularensis 15 NIIEG: order of handling”

DIAGNOSTIC POTENTIAL OF THE ERYTHROCYTIC IMMUNOGLOBULIN DIAGNOSTICUM FOR INDICATION AND IDENTIFICATION OF THE CAUSATIVE AGENTS OF PARTICULARLY DANGEROUS (DEEP) MYCOSES

Objective of the study was to assess analytical and diagnostic sensitivity and specificity of the “Reagent kit. Erythrocytic coccidioidomycosal and histoplasmosal immunoglobulin dry diagnosticum”, designed for identification of causative agents of coccidioidomycosis and histoplasmosis in isolated cultures of micromycetes, as well as in clinical and biological samples using indirect hemagglutination test.Materials and methods. The investigation included 264 positive samples (216 samples of micromycete suspensions, 48 samples of biological and clinical material) containing pathogens of histoplasmosis and coccidioidomycosis concentrated up to 3,12·106 and 1,56·106 cells/ml, respectively, and 128 negative samples containing heterologous microorganisms in concentrations equal to 5·106 cells/ml. The study was carried out using biological samples that were artificially contaminated with stated pathogens of particularly dangerous mycoses and samples, obtained from bioassay animals with experimental infection.Results and conclusions. It is established that diagnostic sensitivity of the reagent kit is not less than 99,0 %. The diagnostic specificity is not less than 98,0 %. Reproducibility of the results in all cases was 100 %. The results obtained testify to the prospect of introduction of the developed kit into the health care practice

Comparison of chromosomal aberrations frequency and polymorphism of GSTs genes in workers occupationally exposed to cytostatics or anaesthetics

Authors compared the incidence of chromosomal aberrations (CAs) of workers occupationally exposed to cytostatics (group EXP1) or anaesthetics (group EXP2) in relationship to polymorphism of GSTM1, GSTP1 and GSTT1 genes. The cytogenetic analysis for chromosomal aberrations frequency and for polymorphisms of genes the PCR and PCR-RFLP method were used. Statistically higher frequency of total CAs was detected in both exposed groups: group EXP1 1.90±1.34%; Mann-Whitney U-test, p=0.001; group EXP2 2.53±1.46%, p=0.0008) as compared to control (1.26±0.93%). In group EXP2 was detected statistically higher frequency of aberrations CSA-type as compared to CTA-type. In xenobiotic metabolizing genes for GST higher frequency of total CAs and constituent types chromatid-type aberrations (CTAs) and chromosome-type aberrations (CSAs) of genes GSTM1 and GSTT1 with null genotype was detected. Statistically significant difference was detected only in CSA-type of aberrations in GSTT1 gene. In gene GSTP1 was not detected any difference in frequency of aberrations in presence of the variant allele. Presented results point out importance of individual susceptibility in evaluation of genotoxic agents of anaesthetics or cytostatics

The thermodynamic properties of the f-elements and their compounds. Part II. The Lanthanide and Actinide Oxides

A comprehensive review of the thermodynamic properties of the oxide compounds of the lan- thanide and actinide elements is presented. The available literature data for the solid, liquid and gaseous state has been analysed and recommended values are presented. In case experimental data are missing, estimates have been made based on the trends in the two series, which are extensively discussed.JRC.E.3-Materials researc