ABSTRACT
AIM: To perform preclinical study of subunit monovalent adsorbed inactivated influenza vaccine "PANDEFLU" [strains A/California/7/2009 (HIN1)v]. MATERIALS AND METHODS: Preclinical study of acute toxicity on experimental animals (assessment of vaccine's toxic effects on organs and body systems; pathomorphologic study of organs and tissues after administration of the vaccine; assessment of its influence on hematologic indicators). RESULTS: It was shown that administration of the vaccine did not lead to death of animals as well as to decrease of body mass or development of pathologic, focal sclerotic changes in parenchymal cells and visceral stroma; the vaccine did not negatively change hematologic and biochemical indicators of blood. Results of necropsy and histological study after acute administration of the vaccine in standard dose did not lead to irritation, inflammation or destruction of tissues in the place of inoculation. The vaccine was apyrogenic and did not have local irritating and allergenic effects. Status of animals after acute inoculation of the vaccine demonstrated its good tolerability and safety in doses exceeding standard human doses more than tenfold. CONCLUSION. Performed research demonstrated absence of contraindications for conduction of clinical trials of "PANDEFLU" vaccine on limited contingent of volunteers.
Subject(s)
Influenza A Virus, H1N1 Subtype/immunology , Influenza Vaccines/adverse effects , Influenza, Human/prevention & control , Animals , Cricetinae , Drug Evaluation, Preclinical , Female , Humans , Influenza Vaccines/administration & dosage , Influenza Vaccines/immunology , Male , Mice , Vaccines, Inactivated/administration & dosage , Vaccines, Inactivated/adverse effects , Vaccines, Inactivated/immunology , Vaccines, Subunit/administration & dosage , Vaccines, Subunit/adverse effects , Vaccines, Subunit/immunologyABSTRACT
The comparative study of adhesive, hemolytic, DNA-ase, lecithinase, antilysozymic, anticomplementary activities of mono- and associated cultures of 57 Enterobacter spp., 61 Citrobacter spp. and 55 Serratia spp. strains, isolated from patients with pyoinflammatory, intestinal and urological diseases is carried out. Different variations of cocultivated bacteria including Enterobacter and Citrobacter, Enterobacter and Serratia, Citrobacter and Serratia are used. It was shown, that cocultivated Enterobacter, Citrobacter and Serratia bacteria increased the persistent properties of mixt cultures.
Subject(s)
Citrobacter/physiology , Enterobacter/physiology , Enterobacteriaceae Infections/microbiology , Serratia/physiology , Animals , Bacterial Adhesion/physiology , Citrobacter/pathogenicity , Complement Inactivator Proteins/metabolism , Deoxyribonucleases/metabolism , Enterobacter/pathogenicity , Enterobacteriaceae Infections/complications , Enterobacteriaceae Infections/pathology , Enterocolitis/microbiology , Hemolysin Proteins/metabolism , Humans , Inflammation/pathology , Muramidase/antagonists & inhibitors , Muramidase/metabolism , Phospholipases/metabolism , Serratia/pathogenicity , Symbiosis , Urinary Tract Infections/microbiology , Urinary Tract Infections/pathologyABSTRACT
During the cultivation of B. subtilis strain 3H under optimum conditions (adequate nutrient medium, seed culture, temperature, the level of dissolved oxygen) protease was produced. Protease could be obtained in the purified form by means of gel chromatography and ultrafiltration. The isolated protease was immobilized on polyglucin and stabilized by intramolecular cross-linking with the use of glutaraldehyde. The comparison of native protease modified with polyglucin and glutaraldehyde, as well as with polyglucin, revealed advantages in the stability of the latter.
Subject(s)
Bacillus subtilis/enzymology , Peptide Hydrolases/isolation & purification , Bacillus subtilis/growth & development , Chromatography, Gel , Culture Media , Dextrans , Enzymes, Immobilized , Glutaral , Hydrogen-Ion Concentration , Oxygen , Peptide Hydrolases/metabolism , Temperature , UltrafiltrationABSTRACT
Proteolytic enzyme produced by Bacillus subtilis is characterized as typical metalloprotease with a molecular weight of 27.9 kD; the enzyme shows its highest activity at pH 7.0-9.0, possesses substrate specificity with respect to different proteins, its temperature optimum is 52 degrees C and its specific activity exceeds that of all known commercial analogues. At 37 degrees C the enzyme is half inactivated in 72 hours.
Subject(s)
Bacillus subtilis/enzymology , Metalloproteases/analysis , Bacterial Proteins/analysis , Bacterial Proteins/isolation & purification , Culture Media , Enzyme Activation , Kinetics , Metalloproteases/isolation & purification , Substrate SpecificityABSTRACT
The effectiveness of the purification of proteolytic enzyme produced by B. subtilis production strain 3H in different nutrient media was evaluated. As revealed in this study, the use of the initial material obtained in peptone nutrient medium made it possible to obtain proteolytic enzyme with the highest specific activity by the methods of salting out, gel and ion-exchange chromatography, but the use of these methods on a production scale led to the deterioration of purity characteristics. Changes in the nutrient medium composition used for the cultivation of the production strain resulted in greater effectiveness of the purification methods. The highly purified preparation of metalloprotease with specific activity exceeding that of available commercial preparations more than twofold was obtained.
