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Biomedical subjects

V A Kolesnikov

Publications and source records attributed to V A Kolesnikov.

At least 19 recordsLinked to original sources
PubMed1991

The delivery of foreign genes into fertilized fish eggs using high-velocity microprojectiles.

Fertilized eggs of loach (Misgurnus fossilis), rainbow trout (Salmo gairdneri) and zebrafish (Brachydanio rerio) were bombarded with high-velocity tungsten microprojectiles covered with plasmid DNA containing sequences of beta-galactosidase and neomycin phosphotransferase genes. About 70% of the eggs survived the bombardment. The activity of both transferred genes was revealed in the fish developed from the bombarded eggs. Neomycin phosphotransferase gene sequences were detected by means of PCR amplification and Southern hybridization in the total DNA of zebrafish that survived after G418 treatment.

Animals↗
PubMed1991

High-velocity mechanical DNA transfer of the chloramphenicolacetyl transferase gene into rodent liver, kidney and mammary gland cells in organ explants and in vivo.

Mouse and rat liver, kidney and mammary gland explants were bombarded with high-velocity microprojectiles carrying a chloramphenicolacetyl transferase gene under different promoters (pTAT-cat, p chi-Casein-cat, p beta-Casein-cat). The expression of a CAT gene was revealed in all organ explants 24 h after transfection. The most pronounced expression was found when a TAT-CAT construction was used. In experiments in vivo rat liver was bombarded in situ with microprojectiles carrying pTAT-cat DNA. A marked activity of the CAT gene was detected 24 h after the bombardment.

Animals↗
PubMed1991

[Changes in the functional activity of blood lymphocytes in patients with destructive pulmonary tuberculosis during the treatment].

Lymphocyte function was studied by the Rigler method modified by V. A. Kolesnikov et. al. in 118 patients with different forms of pulmonary tuberculosis and 15 healthy subjects. This parameter was found to depend on the nature and dynamics of a tuberculosis process and the predominance of the exudative or productive inflammation phase. A considerable reduction of lymphocyte functional activity (LFA) up to 60% was found in the presence of dissemination in pulmonary tissue. Transition from infiltration to destruction was marked by a average rise of LFA from 67 to 79%. The initial LFA parameters can be taken into account as a prognostic criterion. Prescription of immunity stimulators in destructive pulmonary tuberculosis requires LFA evaluation by the above method.

Adjuvants, Immunologic↗
PubMed1991

[Use of high velocity mechanical injection for the transfer of foreign DNA into early mouse embryos].

The possibility of high velocity mechanical transfer of foreign DNA into inner cell mass of mouse blastocyst was shown. Penetration of tungsten microparticles into early embryo cell nuclei and their localization on mitotic chromosomes was demonstrated. About 70% of developing embryos survived the bombardment. Total DNA of the mice born from bombarded embryos was analyzed by blot-hybridization and PCR with Southern hybridization. In three cases, the presence of the transferred plasmid DNA (pSV3-neo) was revealed.

Animals↗
PubMed1990

[High velocity mechanical injection of foreign DNA into fish oocytes].

High-velocity tungsten microprojectiles were used to introduce into fertilized eggs of loach (Misgurnus fossilis), Rainbow trout (Salmo gairdneri Rich.) and Zebra fish (Brachydanio rerio) DNA sequences of beta-galactosidase and neomycin phosphotransferase genes. No more than 30% of fish oocytes died as a result of bombardment. Experiments revealed marked activity of both enzymes in developing fishes. Neo gene DNA sequences were found in total danio DNA using PCR technique.

Animals↗
PubMed1989

Genetic transformation of mouse cultured cells with the help of high-velocity mechanical DNA injection.

NIH 3T3 mouse cells were transfected by the plasmid pSV3neo (G418-resistant) with the help of high-velocity mechanical DNA injection based on the principle of bombarding cells with tungsten particles covered with the DNA. Stable transformants were obtained. Dot-hybridization and Southern analysis revealed the integration into the genome of 5-20 copies per cell of original plasmid DNA. The plasmid DNA was shown to have tandem organization.

Animals↗
PubMed1989

[Computerized tomography in aortic aneurysms].

Experience with computerized tomography, used for the diagnosis of aortic aneurysm, is summarized. This diagnostic method is shown, on the basis of 137 cases, to be highly valuable for this pathology. The results of computerized tomography were compared to postoperative data, which demonstrated high accuracy in the measurement of the size of aortic aneurysm. Being a noninvasive diagnostic method that yields an accurate assessment of the aneurysm size, computerized tomography has advantages over other investigation methods. It is concluded that, unless a combined lesion of other vessels is suspected, preoperative investigation may be limited to computerized tomography alone.

Aorta, Abdominal↗
PubMed1979

[Use of ethidium for the cytochemical study of chromatin].

A cytochemical method of chromatin study by means of nuclear staining with ethidium (bromide) is described. Dependence of stain binding by chromatin on ethidium concentration, ionic composition and buffer pH value has been analyzed. It is suggested that cells be stained in 2.10(-5) M solution of ethidium in 0.1 M tris-HCl buffer at pH 8.0 during 30 min. The fluorescence of nuclei stained with ethidium under conditions described is shown to reflect changes in physico-chemical properties of chromatin taking place in the course of its chemical modification and physiological activation in regenerating liver. The use of ethidium for chromatin cytochemistry allows to study chromatin properties in wide ranges of pH. Some other advantages of the method suggested over the commonly used method of acridine orange staining are discussed.

Acridine Orange↗
PubMed1976

Use of actinomycin D for the specific quenching of fluorescence of deoxyribonucleic acid in cells stained with acridine aminoderivatives.

Actinomycin D specifically quenches the fluorescence of acridine orange and quinacrine bound to deoxyribonucleic acid in cytologic preparations, but does not change the fluorescence of these fluorochromes bound to RNA. The following fluorescence-cytochemical applications of techniques based on these findings can be suggested: (a) distinction between deoxyribonucleic acid and ribonucleic acid; (b) detection of double-stranded virus ribonucleic acid; (c) approximate estimation of the lengths of A-T sequences in deoxyribonucleic acid molecules.

Acridines↗
PubMed1975

Influence of a specific histone kinase on the physico-chemical properties of chromatin in situ.

The influence of a specific histone kinase, phosphorylating lysine-rich histone F1, F2a2, F2b, on the physico-chemical properties of the chromatin in the whole undestroyed fixed cell, has been investigated. It was found that the exogenous histone kinase penetrates into the nuclei of the undestroyed fixed cells and into the isolated unfixed nuclei and changes the physico-chemical properties of the chromatin there, bringing about an increase in binding of a basic dye acridine orange and a decrease in its stability to heat.

Animals↗
PubMedDate not supplied

[Early changes in liver chromatin in response to partial hepatectomy].

Guinea-pig and mouse liver chromatin responds to the partial hepatectomy by an increase in binding of a basic dye acridine orange (AO) and by a decrease of its stability to heat in thermal denaturation test in situ. Degree of the changes in AO chromatin binding is identical in the cells of different ploidy and proportional to their DNA content. Treatment of the preparations by 0.6 M NaCl solutions under conditions bringing about the selective removal of histone H1 from the cells produces in vitro changes in DNA properties taking place in cells in vivo in the course of their activation. The treatment of cells with 0.35 M NaCl solution results in the disappearance of changes occurring in the chromatin of activated cells whereas the properties of control cells remain unchanged. The data obtained are interpreted as a result of the removal of some non-histone regulatory proteins from the chromatin of activated cells that is accompanied by changes in the character of DNA-histone interaction. At the time of maximum increase of AO binding a significant intensification of endogenous RNA polymerase activity was found, the incorporation of [3H] UTP in the nucleolus being higher than that in the extranucleolar part of the nucleus. High ionic strength in the incubation medium (0.4 M (NH4)2SO4) results in drastic increase of radioactive label in the nucleus and in the disappearance of differences between activated and non-activated chromatin. It is concluded that the intensification of RNA synthesis under the influence of proliferative stimulus is more likely dependent on the additional opening of DNA-matrix than on the direct activation of the enzyme.

Animals↗
PubMedDate not supplied

[Influence of histone kinase phosphorylating lysine-rich histones, on the physico-chemical properties of normal hepatocyte chromatin and after partial hepatectomy].

The influence of a specific histone kinase, phosphorylating lysin-rich histone H1, H2a, H2b on the physico-chemical properties of chromatin from hepatocytes of normal and hepatectomized guinea pigs has been investigated. A cytochemical method has been used which permits to obtain information about the physico-chemical properties of the chromatin in situ, i.e. without its isolation. This approach allows us to evaluate changes in chromatin properties in cell cultures as well as in the intact organism. It is found that the specific histone kinase changes the properties of chromatin in non-dividing cells bringing about an increase of acridine orange binding to the level characteristic for hepatocytes after partial hepatectomy. At the same time the chromatin properties in activated hepatocytes are not changed under the action of the histone kinase. It is concluded that the specific histone kinase, phosphorylating lysine-rich histones can play an important role in the course of chromatin activation in cells stimulated to proliferation.

Animals↗