Analysis of DNA polymorphism of cluster genes in cattle: casein genes and major histocompatibility complex (MHC) genes]

The PCR-analysis method has been used to study polymorphism of casein genes (k- and b-caseins) and genes of class of basic histocompatibility complex (DRB and DQB BoLA) in some cattle breeds. Genotyping of k-casein locus has been carried out using analysis of protein polymorphism and DNA polymorphism. It is shown that both methods give identical results and can be used for this purpose, but the PCR-analysis method is more informative. A new allele of k-casein gene (k-Cn F) has been revealed and sequenced using the above method. While analyzing DNA-polymorphism of 5'-nontranslated region of b-casein gene two previously non-described polymorphic sites for restrictase Hinf I are found which occur only in animals of "zebu-like" breed. The high level of polymorphism of bl-domain of gene DRB is shown: polymorphism of bl-domain was revealed already at the level of electrophoretic analysis of PCR-amplification products. For 5'-nontranslated domain gene DQB the polymorphism level was much lower and manifested itself during restriction analysis of amplified fragments.     

Identification of a protein product of a novel human gene SON and the biological effect upon administering a changed form of this gene into mammalian cells

We have investigated the functional activity of human son gene, that possesses the homology to mos and myc genes. Specific antibodies (antiserum) were raised to synthetic peptide, that corresponds to son-protein 943-963 amino acid residues. With this antiserum the presence of son-protein was showed in lysates of cultured human cells transformed by adenovirus type 5, RAT 2 cells and primary human embryonic fibroblasts. son-Protein molecular weight (92 kDa) was determined by the method of electrophoresis in SDS-polyacrylamide gel. Thus, it was shown the presence of son gene protein in animal and human cells. To determine a possible son gene role in mammalian cells we have cloned the 3' part (2667 b.p.) of son cDNA in retroviral vector pPS-3-neo. Transformed cells of different lines were selected. A large portion of this cells changed their morphology. New protein product (120 k), that reacted with antiserum to son specific peptide, was found together with p92son in these clones.     

Characterization of a new hybrid mink-mouse clone panel: chromosomal and regional assignments of the GLO,ACY, NP,CKBB, ADH2, and ME1 loci in mink (Mustela vison)

To expand the mink map, we established a new panel consisting of 23 mink-mouse clones. On the basis of statistical criteria (Wijnen et al. 1977; Burgerhout 1978), we developed a computer program for choice of clones of the panel. Assignments of the following mink genes were achieved with the use of the hybrid panel: glyoxalase (GLO), Chromosome (Chr) 1; acetyl acylase (ACY), Chr 5; creatine phosphokinase B (CKBB), Chr 10; alcohol dehydrogenase-2 (subunit B) (ADH2), Chr 8. Using a series of clones carrying rearrangements involving mink Chr 1 and 8, we assigned the gene for ME1 to the short arm of Chr 1 and that for ADH2 to Chr 8, in the region 8p12-p24. Mapping results confirm the ones we previously obtained with a mink-Chinese hamster panel. However, by means of an improved electrophoretic technique, we revised the localization of the gene for purine nucleoside phosphorylase (NP), which has been thought to be on mink Chr 2. It is reassigned to mink Chr 10.     

Oxidative phosphorylation in membrane vesicles of a gram-positive methylotroph

Membrane preparations, capable of high rates of respiration-linked ATP synthesis, have been obtained from a gram-positive methylotrophic bacterium Bacillus sp. MGA3. NADH, succinate, reduced TMPD and methanol were shown to be suitable substrates for the oxidative phosphorylation. Esterification of orthophosphate was dependent on electron transfer, as evidenced by the requirement for both substrate and oxygen. Phosphorylation was also dependent on ADP and was destroyed by boiling the membrane preparation. The phosphorylation was markedly uncoupled by carbonyl cyanide p-(trichloromethoxy)-phenylhydrazone (CCCP) and was inhibited by N,N-dicyclohexylcarbodiimide (DCCD). KCN caused strong inhibition of substrate oxidation as well as phosphorylation for all substrates tested. Rotenone, amytal and antimycin A caused inhibition when NADH or methanol were used as substrates. Antimycin A inhibited respiration and ATP synthesis with succinate as substrate and had no effect on ascorbate —N,N,N,N-tetramethyl-p-phenylenediimide (TMPD) oxidation by membrane preparations of Bacillus sp. MGA3. P/O ratios determined were 2.4 with NADH, 1.7 with succinate and 0.8 with reduced TMPD. The measured P/O ratio with methanol-oxidizing system was similar to that with NADH (about 2.4).Abbreviations CCCP Carbonyl cyanide p-(trichloromethoxy)-phenylhydrazone - DCCD N,N-dicyclohexylcarbodiimide - TMPD N,N,N,N-tetramethyl-p-phenylenediimide - Q ubiquinone Q     

The use of BrCN for assembling modified DNA duplexes and DNA-RNA hybrids; comparison with water-soluble carbodiimide.

Both cyanogen bromide (BrCN) and 1-ethyl-3-(3'-dimethylaminopropyl) carbodiimide may be used as coupling reagents for the template-directed assembly of DNA duplexes containing the sugar-phosphate backbone modification. Both reagents show similar ligation site structure-specific trend. Practical recommendations are given for selection of the condensing reagent depending on the properties of the duplex. Based on 31P NMR spectroscopy data, a scheme is suggested for BrCN activation of the nucleotide phosphomonoester group. Using both condensing reagents, we studied the condensation of oligonucleotides containing ribo-segments (from mononucleotide residue to full sequence) on the DNA template. Efficiency of the chemical ligation of RNA oligomers was shown to be much lower than that of DNA analogues. The coupling yield depends on the position of the RNA segment in the hybrid duplexes and on the position of the phosphate group in the nick.     

Mechanism of circulatory disorders in animals irradiated at high doses

Some data are reported on pathoanatomical changes, a status of the microcirculatory channel and the coagulogram of animals affected by high doses of ionizing radiation. The signs of disseminated intravascular blood coagulation have been revealed. The authors discuss the relationship between clinical manifestations with coagulopathy development and circulatory disturbances during intestinal and cerebral forms of acute radiation sickness.     

The state of hyporeactivity in interferon production: localization and partial purification of the factor repressing the interferon production

The hyporeactivity factor in interferon production by L-929 cells designated IRP (interferon repressing protein) has been studied. In particular, its localization and methods of its purification have been studied. The kinetics of IRP accumulation by producing cells correlate with the development of hyporeactivity condition. Most of IRP is localized in cell sap and in ribosomal fraction in evidence to regulatory role of repressor at the level of interferon mRNA translation. A 100-fold increase in repressor activity was achieved by IRP concentration by ammonium sulfate precipitation. IRP as well as interferon have been shown to possess high affinity to polyU sepharose. The preparations of IRP and interferon concentrated by ammonium sulfate precipitation were subsequently purified by fractioning in a polyI sepharose column. A 10,000-fold (6 x 10(4) U/mg) purification was achieved for IRP and 250-fold (10(4) U/mg) for interferon.     

Energy and plastic metabolism of the heart muscle in rabbits undergoing thyroid irradiation with decimeter waves

The functional state of myocardial mitochondria, the glycogen and nucleic acid contents in myocardium, and morphometry of structural elements of cardiomyocytes and myocardial capillary network were investigated in order to select the optimal regimen of decimeter wave exposures with power density of 10, 120 and 240 mW/cm2 on the area of thyroid gland. It was shown that the thyroid gland exposure to decimeter waves at these intensities resulted in functional shifts in energy and plastic processes in myocardium and capillary blood supply. These changes increased to a considerable extent as the intensity of exposure was increasing and reached the maximum at power density of 240 mW/cm2 but event at this intensity there was not found the injurious effect of decimeter waves on the myocardium.     

Electron microscopic study of the cytoskeleton of human podocytes

The ultrastructural study of man's cytoskeleton of podocytes is carried out. Populations of podocytes with two different types of structure of the cytoskeleton in dependence on age (2, 4, 6, 37 and 65 years) is revealed in kidneys. The first type of cytoskeleton of the podocyte is peculiar for children's age and is characterized by branched, high density microfilament network, expressed by system of microtubules and single myofilaments. The intermediate filaments here are either utterly absent or present so feebly they find themselves "disguised" by other strongly developed components of cytoskeleton and revealing them with the help of technique of electron microscope is impossible. In kidneys of adults, and especially of old aged persons podocytes with other type of organization of the cytoskeleton are mainly identified. The distinctive signs of the last are bundle arrangement of microfilaments, plural bundles of intermediate filaments and individual microtubules. This study permits to make a conclusion that during individual development and growing old in kidneys of high animals and man, probably, physiological changes causing morphological reconstruction of cytoskeleton which is accompanied by intensive development of intermediate filaments' system with simultaneous "involution" of microtubules and microfilaments' systems take place.