Cytogenetic differences in radiation effects on red-haired field mice from different geographic populations]

The paper presents the data on the effect of X-rays on the incidence of chromosome aberrations and on the mitotic activity of bone marrow cells in two geographical populations of Clethrionomys glareolus. It is shown that the percentage of aberrant cells at different intervals after the irradiation (350 r) is higher in the Moscow population than in the Komi ASSR population. The fractionation radiation results in different increase in the number of aberrant cells. The change of the mitotic activity after X-irradiation was also different in these two geographical populations.     

Isolation and characterization of the ALP1 protease from Aspergillus fumigatus and its protein inhibitor from Physarium polycephalum

It is known that Aspergillus fumigatus secretes a serine protease ALP1 of the subtilisin family in the presence of extracellular protein substrates. We found conditions of A. fumigatus culturing that provide a high ALP1 activity inside cells without induction by extracellular proteins. The identity of the properties of the secreted and intracellular enzymes was shown. A thermostable protein inhibitor of the ALP1 protease was isolated from the plasmodium of the myxomycete Physarum polycephalum. Its molecular mass is 32-33 kDa. The inhibitor inhibits the ALP1 protease activity with IC50 of 0.14 microM. This protein was also shown to be a less efficient inhibitor of the activity of HIV-1 protease (IC50 2.5 microM). The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 3; see also http://www.maik.ru.     

Molecular cloning,expression and characterization of human mini antibodies against enterotoxin C1 of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis>

We describe here the cloning, expression, and production of specific single-chain antibodies (scFv) against the recombinant enterotoxin C1 of Staphylococcus aureus. High-affinity scFv were selected from the phage library of human mini antibodies; afterwards, the cells of E. coli trxA gor double mutant were infected with a product obtained by fusion of DNA encoding of these mini antibodies with the trxA gene to induce soluble scFv synthesis in cell cytoplasm. The scFv obtained displayed high enterotoxin C1 affinity. Analysis for cross reactivity showed that mini antibodies interacted also with SEA- SEB-, SED-, SEE-, SEG-, and SEI-type enterotoxins, but they failed to interact with ricin, diphtheritic, and cholera toxins, or with both lethal and protective factors of the anthrax toxin. This property may be helpful in screening for staphylococcus enterotoxins.     

Preparation and characterization of monoclonal antibodies to the cholera toxin

Monoclonal antibodies to cholera toxin were obtained. They do not cross-react with the termolabile toxin (LT) of Escherichia coli, ricin, diphtherial toxin, staphylococcus enterotoxins of SEA, SEB, SEI, SEG, or the lethal factor and protective antigen of the anthrax toxin. Pairs of antibodies for the quantitative measurement of the cholera toxin in sandwich enzyme immunoassay (EIA) were selected. The detection limit of the toxin is 0.2 ng/ml for plate EIA and 0.44 ng/ml for microchip EIA. The presence of milk, broth, or surface water in the toxin samples does not reduce the sensitivity of EIA.     

Bacteriorhodopsin Retains the Conformation of Val69–Gly72 Fragment during Functioning

Effect of the monoclonal antibody (MAb) 5B6 produced to the solubilized preparation of bacteriorhodopsin on the protein photocycle was studied to examine conformational rearrangements on the surface of a functioning bacteriorhodopsin molecule. Using solid phase enzyme immunoassay, peptide phage display, and ¹H NMR spectroscopy, we demonstrated that the epitope recognized by MAb 5B6 is the Val69-Pro-Phe-Gly72 fragment of the protein, with the aromatic ring of Phe71 and the methyl groups of Val69 participating in the binding. MAb 5B6 exerted no significant effect on the photocycle of bacteriorhodopsin solubilized in Triton X-100 at pH 6.2 and 7.4 which suggested that, when functioning, bacteriorhodopsin retains the conformation and position of its Val69-Pro-Phe-Gly72 fragment.     

Bacteriorhodopsin retains the conformation of Val69-Gly72 fragment during functioning

Effect of the monoclonal antibody (MAb) 5B6 produced to the solubilized preparation of bacteriorhodopsin on the protein photocycle was studied to examine conformational rearrangements on the surface of functioning bacteriorhodopsin molecule. Using the methods of solid phase enzyme immunoassay, peptide phage display, and 1H NMR spectroscopy, we demonstrated that the epitope recognized by MAb 5B6 is the Val69-Pro-Phe-Gly72 fragment of the protein, with the aromatic ring of Phe71 and the methyl groups of Val69 participating in the binding. MAb 5B6 exerted no significant effect on the photocycle of bacteriorhodopsin solubilized in Triton X-100 at pH 6.2 and 7.4, which suggested that, when functioning, bacteriorhodopsin retains the conformation and position of its Val69-Pro-Phe-Gly72 fragment.     

Evaluation of immunobiological activity of Francisella tularensis C-complex preparations as promising component of subunit vaccines

Data on influence of Francisella tularensis C-complex preparations on formation of immunity against tularemia are presented. Study of cellular immunity characteristics as well as dynamics of antibody response was carried out on white mice and guinea pigs models. Absence of toxicity, pyrogenicity, and negative effects on immunocompetent cells in combination with protective activity points to possibility of use the C-complex as a component of a subunit vaccine.     

New hematological method for detecting long-term consequences of short-term myocardial ischemia

It has been shown in chronic experiments on dogs with reversible disorders of the coronary circulation that the sequels of a 3-5-minute reversible myocardial ischemia might be identified throughout 24-120 hours by an original method suggested by the authors. The method is based on the diagnosis of latent postischemic leukocytosis and other associated hematological shifts.