[Humoral factors of immunity and the mononuclear cell responses to fetoproteins in patients with pulmonary tuberculosis].

The content of cytokines, the levels of antibodies (Ab) to proinflammatory cytokines and fetoproteins (FP) in serum, as well as the effects of fitohemagglutinin and FP on the level of cell production of cytokines into the conditioned medium were studied in relation to the pattern of a response of mononuclear cells (MNC) to FP. With the positive reaction of MNC to FP, estimated by an increment of CD9* cells and detectable in grades 2-3 dysplasias, the anti-inflammatory effect with lower anti-inflammatory cytokines was shown to be achieved due to elevated Ab levels, which may compensate for the low content of IL-4 and IL-10. With FP, there was an increase in the cell production of IL-10 that is known to stimulate antibody formation in the early phase of tumor growth, as evidenced by the association of the levels of Ab to FP with the grade of dysplasia.

[L-Glutamic acid modulates the cytotoxic effect of tumor necrosis factor in the HL-60 cell line].

L-Glutamic acid was shown to increase the stability of cells of the HL-60 line of human promyelocyte leukemia to the cytotoxic action of tumor necrosis factor alpha (TNF-alpha) due to the inhibition of apoptotic and NF-kappaB-activating cascades induced by this cytokine. At the same time, L-glutamic acid increases the TNF-alpha-mediated differentiating signal and the accompanying enhancement of the phosphatidylinositol-specific phospholipase C activity. Therefore, it is a promising agent for the reduction of total toxicity and inflammatory processes during treatment with TNF-alpha.

[Results of Befnorin clinical trails in healthy volunteers].

Clinical trails of Befnorin based on the human recombinant TNF-beta elaborated at the Research Design and Technology Institute of Biologically Active Substances, "Vector" State Research Center of Virology and Biotechnology, were carried out on healthy volunteers in compliance with a decision passed by the Committee of Medical and Immunobiological Preparations, Russia's Health Ministry. Single Befnorin doses of 5-10(4) U, 10(5) U, 5-10(5) U, and 10(6) U were administered as intramuscular injections. Clinical, biochemical and immunological parameters were registered for 7 days after a single dose.

[Effect of tumor necrosis factor alpha on the efficiency of anti-rabies vaccination].

Recombinant tumor necrosis factor-alpha (TNF-alpha) improved the survival of random-bred albino mice vaccinated with antirabies vaccine after infection with rabies CVS strain. The agent dose of 0.1 microgram/animal, injected 1 day postvaccination, was the most effective.

[Effect of human recombinant granulocyte colony-stimulating factor on mouse hematopoietic and lymphoid organs].

Experiments on intact CBA/CALac mice demonstrated that single (1.25 mg/kg) and repeated (0.125 and 1.

[The determination of lipopolysaccharide admixtures in protein preparations].

A new method for the determination of lipopolysaccharide (LPS) admixtures in protein solutions has been developed. The method includes the periodate oxidation of LPS, biotinylation with biotin hydraside, immobilization on a nitrocellulose membrane and the development of biotinylated LPS in the streptavidin--alkaline phosphatase system. Proteins are previously removed from the solution by treatment with hot phenol.

[The immunoregulatory properties of recombinant human tumor necrosis factor-beta in mice opposite-reacting to antigen].

The effect of 10(3)-10(5) E/20 g doses of the recombinant factor of human beta tumor necrosis (rFNT-beta) on formation of the immune response and macrophage functional activity was studied in CBA and C57Bl/6 mice that differ in genetically determined level of the immune response to an antigen (sheep erythrocytes). The rFNT-beta was found to cause a modulating effect on the cell and humoral links of the immune response. The effect of the agent depended on the dose and the genotype of the experimental animals.

[The blood-simulating properties of a recombinant colony-stimulating factor and glycyram during cytostatic myelosuppression].

The hemostimulating activity of the drug glycyrram and the recombinant granulocytic-macrophagic colony-stimulating factor (CSF) was compared on models of myelosuppression induced by injection of an alkylating agent cyclophospane (CF) or the antimetabolite 5-fluorouracil (5-FU). It was found that glycerram stimulated hematopoiesis more effectively after 5-FU injection but to a lesser extent (in relation to CSF) in CF injection.

[Study of antiinfectious potential of recombinant tumor necrosis factor-alpha].

Tumor necrosis factor-alpha (TNF) was shown to have an antiinfectious effect on murine and rat models of viral, bacterial and parasitic infections. The effective antiinfectious dose of TNF was determined to be 10(2)-10(3) U/kg body weight. TNF, interferon, and antibiotics were demonstrated to potentiate the antibacterial effect of each.

[Design of stimulating agents of non-specific resistance system].

The paper briefly reviews a study on the design of drugs enhancing the body's nonspecific resistance to pathogenic agents. To examine the potential regulatory effects on cytokine function was a main trend. The interferon inductor ridostin, dsRNA of microbiological origin, cytokines, as well as the recombinant probiotic strain yielding interferon alpha-2 synthesis were used as a pharmacological agent.

[Preparation of lymphotoxin and properties of it].

The process of isolation of highly purified human lymphotoxin was studied and optimized. A set of methods providing an increase in the content of the target product in the biomass (plasmid DNA amplification and selection of clones of transformed cells) was applied at the stage of cultivation. A two-step purification scheme (ion-exchange chromatography on DEAE-Biotone A1 and hydroxylapatite) was developed.

[Toxic properties of recombinant tumor necrosis factor].

Toxic properties of recombinant human tumor necrosis factor (TNF-beta) were studied on noninbred albino mice. In the maximum tolerable doses the preparation induced a decrease in the body weight and temperature of the animals as well as development of glyco- and leukopenic reactions and damage of the internal organ structures. The preparation effects were observed early after the exposure and were mainly reversible.

[Biosynthesis of alpha2-interferon and tumor necrosis factor at different stages of culturing].

Study of the biosynthesis of human alpha 2-interferon and tumor necrosis factor in derivatives of E. coli SG200-50 recipient strain containing pIF16, pTNF311, and pLT21 plasmids demonstrated elimination of the plasmids from the cells without degradation thereof in the course of culturing. Methods increasing the content of the end products in the biomass are proposed.

[Preparation of a mutant human tumor necrosis factor with increased resistance to proteases].

The potential proteolysis sites of human TNF are considered. By site-directed mutagenesis the Arg-31 residue of mature TNF was substituted by Gln. The analysis of cytotoxicity of initial and mutant (R31Q) proteins on mouse L929 fibroblasts did not reveal any differences in biological activity.

[Preparation and bacterial expression of a mutant gene for human lymphotoxin].

Using the oligonucleotide directed mutagenesis, a human lymphotoxin (TNF beta) mutant gene lacking 21 N-terminal codons has been obtained. Recombinant plasmid pLT21 for expression of the mutant gene has been constructed. The mutant gene in the plasmid was placed under control of a tandem of constitutive promoters from coliphage T7.

[BME 361 I--a new site-specific deoxyribonuclease type II from Bacillus megaterium 361].

Bme 361 I, a new site-specific type II deoxyribonuclease, was purified from Bacillus megaterium 361 by chromatography on phosphocellulose P 11 and hydroxylapatite. The enzyme recognizes and cleaves the nucleotide sequence 5'-GG decreases CC-3' in double-strand DNA. Thus it is a true isoschizomer of deoxyribonucleases Hae III and BspR I.

[Restriction endonucleases: the detection of new producer strains and the isolation of enzyme preparations].

The method for analysis of microorganisms for the presence of the modification-restriction systems has been developed. The method has permitted to detect more than 10 new producing strains of restrictases including microorganisms of Rhizobium genus. Some of them are promising for practical use.

[Site-specific restriction endonuclease RME211 from Rhizobium meliloti].

A new site-specific restriction endonuclease Rme211 from Rhizobium meliloti has been shown to recognize the following nucleotide sequence 5'-ATCGAT-3' in the double-stranded DNA. Thus, the enzyme is a true isoschizomer for restriction endonucleases Bsu151 and ClaI.

[Isolation of SAU 96 I restrictase, its physical and catalytic properties].

Restrictase Sau 96 I was isolated from Staphylococcus aureus PS 96 and purified by chromatography on DEAE-cellulose, phosphocellulose and hydroxylapatite. The preparation was studied by gel filtration on Toyopearl HW-55 and polyacrylamide gel electrophoresis under denaturing conditions. The active form of the enzyme is a dimer with a molecular weight of 54,000 +/- 5000 composed of two identical subunits.

[Substrate specificity of T4 RNA-ligase. The effect of the nucleotide composition of substrates and the size of phosphate donor on the effectiveness of intermolecular ligation].

Nucleoside 2' (3'),5'-diphosphates, dinucleotides pApA, pApC, pApU, pGpC, pCpC, pUpU (phosphate donors), and trinucleoside diphosphates, such as NpCpC, NpCpU, NpUpC, NpUpU and GpApN (N = U, C, A or G; phosphate acceptors) were used to study the substrate specificity of T4 RNA ligase. Relative efficiency of the mono- and dinucleotide donors depends on the 5'-terminal nucleoside moiety of the dinucleotide: upon ligation with the minimal phosphate acceptor GpUpC, dinucleotides pApA, pApC, and pApU are more effective than nucleotide diphosphate pAp; pGpC is more effective than pGp; efficiencies of pCpC and pCp are almost identical, and efficiency of pUpU is slightly lower than that of pUp. In relative efficiency, dinucleotide donors, varying only in 5'-terminal unit, do not correspond to mononucleotides: pApC greater than pCpC greater than pGpC and pCp greater than pUp approximately pAp much greater than pGp.