Effect of brain and thymus cationic proteins on membrane permeability]

Cationic proteins of brain lysosomes (LCP), myelin (MCP) and nuclear histone fractions from calf thymus (T) and rat brain (B) are shown to increase at different degree the permeability of brain lysosomes and neutrophiles for acid RNAase, acid phosphatase, catepsin D and beta-galactosidase. According to the effectivity, basic proteins can be listed in the following order: for lysosomes-f2aT, F3B, f3T greater than total histones B, f2bT greater than f2B greater than LCP, MCP greater than flT, flB; for neutriphiles-f3T larger than or equal to total histones B larger than or equal to f3b MCP larger than or equal to f2aT, f2bT greater than f2B greater than LCP greater than flB greater than flT. Fractions f2a and f3 considerably increased the release of acid RNAase from lysosomes in very low concentrations beginning from 0,2 mug/ml, while the release of catepsine and acid phosphatase took place beginning from 5-10 mug/ml. The effect of lysosome and myelin cationic proteins on the release of hydrolases occurred at concentrations ten to hundred times higher.     

Results of phase I clinical trials of a combined vaccine against HIV-1 based on synthetic polyepitope immunogens

The CombiHIVvac candidate vaccine against HIV-1/AIDS containing two synthetic polyepitope immunogens such as TBI and TCI to stimulate the humoral and cellular response is described. The recombinant TBI protein is constructed as a polypeptide with predetermined tertiary structure and contains epitopes of Env and Gag proteins of HIV-1. TCI contains CD8⁺ CTL and CD4⁺ Th epitopes of the major viral proteins such as Env, Gag, Pol and Nef which are highly conserved among subtypes A, B and C of HIV-1. A gene encoding the polyepitope TCI immunogen is inserted into a pcDNA-3.1 plasmid vector. The CombiHIVvac vaccine was designed as virus-like particles containing the pcDNA-TCI plasmid in their cores (DNA vaccine) and the TBI protein conjugated with polyglucin on their surfaces. Immunogenicity and safety of CombiHIVvac has been shown in preclinical studies in several animal species. Phase I clinical trials of the vaccine have been completed and the results obtained in human volunteers confirmed that the CombiHIVvac candidate vaccine was safe and did not cause side effects, at the same time, inducing the HIV-specific humoral and cellular immune response. The phase II clinical trials have been approved by the Ministry of Health and Social Development of the Russian Federation.     

Cooperation of Hepatocytes in vitro in the Rhythm of Protein Synthesis is Promoted by the GM1 Ganglioside Contained in Vesicles and Liposomes

The medium conditioned by dense, self-synchronized hepatocyte cultures was centrifuged at 150000 g to obtain two fractions. The light fraction (supernatant fluid) contained ganglioside monomers and micelles, and the heavy fraction (pellet) contained gangliosides in the vesicles shed from the cell membrane. In the test populations of hepatocytes, the rhythm of protein synthesis was used as an indicator of cell synchronization resulting from their cooperative activity. Diluted hepatocyte cultures with asynchronous fluctuations of protein synthesis proved to be synchronized by both the initial conditioned medium and its vesicular fraction. Our previous studies have shown that this occurs under the effect of GM1 monosialoganglioside, which is released from cultured cells and accumulated in the conditioned medium. Liposomes consisting of GM1 and phosphatidylcholine from egg yolk (1 : 19 mol%), compared to free exogenous GM1, synchronized the rhythm of protein synthesis more effectively: synchronization was observed at a GM1 concentration in liposome suspension of only 0.0003 M, compared to 0.06 M and higher in the case of free GM1. Thus, GM1 as a component of membranes and monolayer lipid structures proved to be much more effective than free GM1 in promoting hepatocyte cooperation with respect to the rhythm of protein synthesis.     

Isolation of Escherichia coli K12 mutants with deficient in precise excision of transposons

Plasmid pNM1, the derivative of R100.1, has been constructed by insertion of transposon Tn5 into structural tet genet (Tn10) of the parental plasmid. The frequency of precise excision of Tn5 from plasmidic genome is 10(-5). The high frequency of precise excision obtained in this system permits one, to use it for isolation of mutants having low frequencies of precise excision. Two mutants were isolated in which the frequencies of precise excision of Tn5 were decreased for two orders. The pex1 and pex2 mutations responsible for the effect decrease the precise excision of Tn5 from R100.1 as well as from RP4 genomes.     

Kanamycin levels in treating patients with acute and subacute gonorrhea

Kanamycin levels in blood serum were studied during treatment of patients with acute and subacute not complicated gonorrhea. It was concluded that in treatment of gonorrhea kanamycin should be administered intramuscularly in a dose of 1 g every 12 hours or in a dose of 0.5 g every 6 hours, the course dose being 3 g.     

Characteristics of continuous cell line 293 used for preparation of antineoplastic agent Cancerolysin]

Certification of continuous cell 293 culture used for cultivation of antineoplastic preparation Cancerolysin was carried out. The seeding and working banks of cells 293 were established and deposited for storage at the Vector Centre. The cells were certified in accordance with the WHO requirements. The cell 293 culture was shown to have high proliferative activity; morphology typical of the line; its karyotype and enzymogram are typical of human cells; the culture is not contaminated with bacteria, fungi, mycoplasms and viruses including oncogenic ones; it has high virus-producing activity; it preserves stability of all the biological properties in long-term cultivation. The seeding and working cell banks were recommended for the use in production of drugs for the treatment of oncologic patients.     

Adverse effects of tempol on hidrosaline balance in rats with acute sodium overload

We studied the effects of tempol, an oxygen radical scavenger, on hydrosaline balance in rats with acute sodium overload. Male rats with free access to water were injected with isotonic (control group) or hypertonic saline solution (0.80 mol/l NaCl) either alone (Na group) or with tempol (Na-T group). Hydrosaline balance was determined during a 90 min experimental period. Protein expressions of aquaporin 1 (AQP1), aquaporin 2 (AQP2), angiotensin II (Ang II) and endothelial nitric oxide synthase (eNOS) were measured in renal tissue. Water intake, creatinine clearance, diuresis and natriuresis increased in the Na group. Under conditions of sodium overload, tempol increased plasma sodium and protein levels and increased diuresis, natriuresis and sodium excretion. Tempol also decreased water intake without affecting creatinine clearance. AQP1 and eNOS were increased and Ang II decreased in the renal cortex of the Na group, whereas AQP2 was increased in the renal medulla. Nonglycosylated AQP1 and eNOS were increased further in the renal cortex of the Na-T group, whereas AQP2 was decreased in the renal medulla and was localized mainly in the cell membrane. Moreover, p47-phox immunostaining was increased in the hypothalamus of Na group, and this increase was prevented by tempol. Our findings suggest that tempol causes hypernatremia after acute sodium overload by inhibiting the thirst mechanism and facilitating diuresis, despite increasing renal eNOS expression and natriuresis.