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Biomedical subjects

V A Kabanov

Publications and source records attributed to V A Kabanov.

At least 19 recordsLinked to original sources

Pluronic micelles as a tool for low-molecular compound vector delivery into a cell: effect of Staphylococcus aureus enterotoxin B on cell loading with micelle incorporated fluorescent dye.

Micelles of pluronic P85 (poly(oxyethylene)-poly(oxypropylene) block copolymer) are used as microcontainers for in vitro delivery of fluorescein into Jurkat and MDCK cells. In order to target the fluorescein containing micelles into the cell, Staphylococcus aureus enterotoxin B (SEB) is covalently conjugated with a pluronic molecule and the conjugate is incorporated into the micelle content. The incorporation of SEB capable of receptor-mediated endocytosis results in a drastic enhancement of the efficiency of cell loading with the fluorescent dye. This effect is not observed under the conditions (4 degrees C) when endocytosis is abolished.

Animals

DNA interpolyelectrolyte complexes as a tool for efficient cell transformation.

A tool was developed for enhancement of plasmid penetration into an intact cell, based on increasing DNA hydrophobicity via inclusion into a soluble interpolyelectrolyte complex (IPC) with polycations. The characteristics of formation of DNA IPC with synthetic polycations [poly(N-ethyl-4-vinylpyridinium)bromide (PVP) and PVP modified with 3% of N-cetyl-4-vinylpyridinium units (PVP-C)] were studied using ultracentrifugation and polyacrylamide gel electrophoresis methods. The conditions were established under which the mixing of DNA and polycation aqueous solutions results in the self-assembly of soluble IPC species. Incorporation of DNA into IPC results in the enhancement of DNA binding with isolated Bacillus subtilis membranes. A considerable increase in the efficiency of transformation of B. subtilis cells with pBC16 plasmid resulted from incorporation of the plasmid into the IPC with PVP and CVP.

Bacillus subtilis

Recognition of target colloid species by conjugates of a linear polyelectrolyte with a vector protein.

Redistribution reaction of quaternized poly-4-vinylpyridine polycations and their conjugates with alpha-chymotrypsin by oppositely charged latex particles is disclosed. The polycations are strongly adsorbed on the latex surface. Nevertheless, they are able to migrate between the latex species via occasional interparticle contacts. In the case of a homogeneous latex such interchange results in uniform distribution of polycations by latex particles. The distribution drastically changes, when alpha-chymotrypsin-polycation conjugates interact with a mixture of two latexes: one chemically modified by bovine serum albumin and the other one by specific protein inhibitor of alpha-chymotrypsin. In this case the interchanging polycations are finally fixed on the latex particles carrying the centres of specific binding of the enzyme vector, i.e. recognize them in the latex mixture. The obtained results are considered to mimic physico-chemical interaction and recognition of target supermolecular bio-objects by large macromolecules carrying relatively small molecular vectors.

Adsorption

Respecrins--a new type of compound with targeted action.

A new technique for the creation of compounds with targeted action is described. In such compounds, termed respecrins (receptor-specific, screened toxin), a physiologically active component carries an epitope-containing fragment of the target antigen and is screened or masked by antibody specific to this antigen. Interaction of the respecrin with the target cell results in dissociation of this immunocomplex, and thereby activation of the physiologically active component. The kinetics of dissociation of a respecrin during interaction with a cell were investigated by flow cytometry. Possible applications of the principle are illustrated by investigation on the antigen-dependent activation of the mitogenic activity of staphylococcal enterotoxin A.

Allosteric Regulation

[Plastic surgery using cutaneo-fascial flaps in the treatment of burns].

Results of 67 plasties with fasciocutaneous flaps in 59 patients operated upon for burns and their sequellae are described. The correlation of length and width of the flaps was from 1.5:1 and 3:1. Good results were obtained in 90% of the patients. Causes of unfavorable outcomes are analyzed. The method is recommended for the introduction into practice of treatment of burns and their sequellae.

Adolescent

Enzymes in polyelectrolyte complexes. The effect of phase transition on thermal stability.

Penicillin amidase, alpha-chymotrypsin and urease have been immobilized in water-soluble nonstoichiometric polyelectrolyte complexes (N-PEC). N-PEC are formed by modified poly(N-ethyl-4-vinyl-pyridinium bromide) (polycation) and excess poly(methylacrylic acid) (polyanion). N-PEC are a new class of polymers capable, characteristically, of phase transitions solution in equilibrium precipitate induced by slight change in pH or ionic strength. Neither the chemical structure of the carrier nor the number of cross-linkages between an enzyme and a carrier change on phase transition. That gives an unique opportunity to elucidate the difference between enzymes immobilized on water-soluble and water-insoluble supports. A detailed study of the phase transition effect on thermal stability of the enzymes and protein-protein interactions has been carried out. The following effects were found. Pronounced thermal stabilization of penicillin amidase and urease may be achieved on two conditions: the enzyme is in the precipitate; (b) the enzyme is linked to the N-PEC nucleus. Then the thermal stability of N-PEC-bound penicillin amidase increases 7-fold at pH 5.7, 60 degrees C, and 300-fold at pH 3.1, 25 degrees C, compared to the native enzyme. For urease, the thermal stabilization increases 20-fold at pH 5.0, 70 degrees C. The localization of enzyme on N-PEC has been established by titration of alpha-chymotrypsin bound to a polycation or polyanion with basic pancreatic trypsin inhibitor. Both in solution (pH 6.1) and in N-PEC precipitate (pH 5.7), an alpha-chymotrypsin molecule bound to a polyanion is fully exposed to the solution. If the enzyme is bound to a polycation, only 20% of alpha-chymotrypsin molecules in the precipitate and 40% in solution retain their ability for protein-protein interactions. This means that a polycation-bound enzyme is localized in the hydrophobic nucleus of the complex, whereas the polyanion-bound enzyme sits on the hydrophilic shell of the complex. On pH-induced phase transition (pH decreases from 6.1 to 5.7), there occurs a stepwise decrease in penicillin amidase activity which is due to a 9.8-fold increase in the Km for 2-nitro-4-phenylacetamidobenzoic acid. Change of the catalytic activity and thermal stability of N-PEC-bound penicillin amidase is fully reversible and reproducible. Such soluble-insoluble immobilized enzymes with controllable thermal stability and activity may be used for simulating events in vivo and in biotechnology.

Chymotrypsin

[Enhancement of the immune response to surface antigens of the influenza B virus as a result of their covalent binding with a synthetic polymer carrier].

The primary immune response (the number of antibody-forming cells, AFC) and the delayed type hypersensitivity (DTHS) were studied in mice immunized either with isolated glycoproteins of influenza virus (hemagglutinin, HA and HA plus neuraminidase, HA plus NA) or with their conjugates with an acrylic acid copolymer (CP) and N-vinylpyrrolidone of equimolar composition. Immunization of mice with conjugates containing virus proteins (virogates-HA-CP or HA plus NA-CP--entailed a 50-100 increment of the number of IgM- and IgG-AFC, anti-HA as compared with analogous parameters during immunization of animals with isolated virus proteins. Immunization of mice with the virogate HA-CP gives rise to the development of a more pronounced DTHS to HA. The authors discuss the possibility of the use of this basically new approach to the design of highly immunogenous vaccine preparations, effective in the control of influenza and other virus diseases.

Acrylic Resins

[Mechanisms of action of synthetic polyelectrolytes and polyampholytes on the immune response].

We investigated the effect of polyacrilic acid (PAA) on the immune response in mice of various strains on sheep red blood cells and also the influence of poly-2-methyl-5-vinyl-pyridine (PMVY), PAA and their statistical copolymers on antibody-forming cells (AFC) production in cultures of T- and B-lymphocytes in vivo. PAA was seen to increase accumulation of AFC in the spleen of mice depending on their genotypes. PMVP and PAA were found to intensify the cooperating interaction of T- and B-lymphocytes, whereas their copolymers exert quite an opposite effect. The injection of copolymers to the recipients of cooperating T- and B-lymphocytes practically results in the complete elimination of the cooperation effect between T- and B-lymphocytes in the immune response to sheep erythrocytes without cytostatic action of cell proliferation.

Acrylic Resins

[Cooperative interaction of serum albumin with quaternized poly-4-vinyl pyridine and structure of the complexes].

Interaction of bovine serum albumin (BSA) with quaternized poly-4-vinyl pyridine (PE) in aqueous solutions at pH 7 was studied. It was shown that in a wide range of the ratios of the components (nBSA/nPE) soluble stable cooperative complexes were formed. At the same time a certain critical content of the protein exists at which the system loses its homogeneity. Complex formation is not accompanied by protein denaturation. At smaller nBSA/nPE ratios non-homogeneous distribution of protein globulas among polyelectrolite macromolecules was found; this corresponded to the "all or none" principle. Using ultracentrifugation technique viscosimetric measurements and electron microscopy it was shown that the soluble complexes exist in the form of rode-like particles consisting of protein globules stabilized by polycation chains. Such particle can be considered as a model of nucleoprotein complex. At certain crytical nBSA/nPE rations the rod-like particles aggregate with additional number of BSA-molecules and form more complicate soluble and insoluble cooperative complexes. Possible structural models of the complexes described were suggested and the thermodinamic and kinetic cryteria of their self-assembly were discussed.

Drug Stability

[2 types of soluble complexes of bovine serum albumin with polyelectrolytes].

It has been shown by high-velocity sedimentation that BSA forms soluble complexes with polyacrylic acid (PAA) and sodium polystyrene sulfonate (PSSNa) with neutral water solutions. Factors responsible for polyanion binding of similarly charged BSA molecules are analysed. It has been found that occupation of PAA and PSSNa chains with BSA globules proceeds steadily. BSA-PAA and BSA-PSSNa complexes are compared with the earlier described products of BSA sorption on poly-4-vinyl-N-ethylpyridine bromide, where the occupation of polycations with protein globules proceeds unevenly. Reasons for the existence of two types of soluble BSA-polyelectrolyte complexes are described; probable structural model of BSA complex with polyacids and polyanions is suggested.

Acrylic Resins