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Biomedical subjects

V I Baranov

Publications and source records attributed to V I Baranov.

At least 19 recordsLinked to original sources

[The oxygen diffusion coefficient in isolated skeletal muscle fibers].

The oxygen diffusion coefficients in isolated skeletal muscle fibres of Baikal seal and grayling, white rat, chicken were calculated on the basis of experimentally determined curve of dependence of the O2 consumption on pO2 and the radius of the fibres. In all the cases, the diffusion coefficient was smaller in the fibres than in water. The oxygen diffusion coefficient was increased during embryogenesis and postnatal period in chicken. The influence of different parameters of the oxygen transport and of properties of different fibres on the values of the oxygen diffusion coefficients, are discussed.

Animals

[Determining the level of yttrium oxide-coated silicon nitride in the air of the work area].

To measure the concentrations of yttrium-oxide-coated small-dispersed silicon nitride plasma powder, the air was aspirated at the amount of 10-15 litres per min through the AFA-XP filter. The filter was limed in a platinum crucible, the residuum being alloyed with Na2CO3, Na2B4O7, NaNO3 in ratio 10:5:1. The alloy was transferred into solution in which the silicon content was assessed through reactions with ammonium molybdate, and yttrium--through reactions with arsenaso III. The sensibility rate for silicon was at 0.07 mg/m3.

Air Pollutants, Occupational

Gene expression in a cell-free system on the preparative scale.

A cell-free system for preparative gene expression is described. It is composed of DNA-free Escherichia coli extract and added plasmid DNA; coupled transcription-translation proceeds with a continuous flow of the feeding solution containing nucleoside triphosphates and amino acids. The system works at a high constant rate for tens of hours. The yield of synthesised proteins after 20-50 h is hundreds of micrograms from 1 ml of the reaction mixture. Electrophoretic analysis of translation products confirms synthesis of proteins of the expected molecular mass.

Adenosine Triphosphate

A continuous cell-free translation system capable of producing polypeptides in high yield.

A cell-free translation system has been constructed that uses a continuous flow of the feeding buffer [including amino acids, adenosine triphosphate (ATP), and guanosine triphosphate (GTP)] through the reaction mixture and a continuous removal of a polypeptide product. Both prokaryotic (Escherichia coli) and eukaryotic (wheat embryos, Triticum sp.) versions of the system have been tested. In both cases the system has proven active for long times, synthesizing polypeptides at a high constant rate for tens of hours. With the use of MS2 phage RNA or brome mosaic virus RNA 4 as templates, 100 copies of viral coat proteins per RNA were synthesized for 20 hours in the prokaryotic or eukaryotic system, respectively. With synthetic calcitonin messenger RNA, 150 to 300 copies of calcitonin polypeptide were produced per messenger RNA in both types of continuous translation systems for 40 hours.

Bacteriophages

Does the channel for nascent peptide exist inside the ribosome? Immune electron microscopy study.

MS2 phage RNA-directed synthesis of an N-terminal polypeptide of the phage coat protein on Escherichia coli 70 S ribosomes was initiated in a cell-free system with the N-dinitrophenyl derivative of methionyl-tRNAFMet) and performed in the absence of tyrosine, lysine, cysteine and methionine. As a result, the translating ribosomes carried peptides up to 42 amino acid residues in length with the dinitrophenyl hapten at the N-ends. Using the immune electron microscopy technique the positions of the nascent peptide N-ends on the 70 S ribosomes have been visualized. It has been found that (i) the N-ends of nascent peptides of these lengths are accessible to antibodies, (ii) the exit site of a nascent peptide is the pocket between the base of the central protuberance and the L1 ridge on the 50 S subunit, i.e. presumably its peptidyl transferase center, and (iii) the further pathway of a nascent peptide seems to proceed along the groove on the external surface of the 50 S subunit.

Antibodies

Is the three-site model for the ribosomal elongation cycle sound?

The release of deacylated tRNA from the ribosome as a result of translocation has been studied. Translating ribosomes prepared with poly(U)-S-S-Sepharose columns have been used. It has been shown that deacylated tRNA released from the ribosomal P site as a result of translocation rebinds with the vacated A site. Consistent with the known properties of the A site of the ribosome, this interaction is reversible, Mg2+-dependent, codon-specific and is inhibited by the antibiotic tetracycline. It has been concluded that the proposed three-site model of the ribosomal elongation cycle (Rheinberger and Nierhaus (1983) Proc. Natl. Acad. Sci. USA 80, 4213-4217) is not sound: the experimentally observed 'retention' of the deacylated tRNA on the ribosome after translocation can be explained by a codon-dependent rebinding to the A site, rather than by its transition to the 'E site', i.e., in terms of the classical two-site model.

Models, Biological

Translocation makes the ribosome less compact.

Translating ribosomes of Escherichia coli were prepared either in the pre-translocation or in the post-translocation states by a special technique based on the use of poly(U)-Sepharose columns where the template was coupled to the matrix through splittable -S-S- bridges. Elongation factors were absent from the final preparations. A neutron scattering study of the translating ribosomes in the two functional states was performed at different contrasts (various 1H2O/2H2O mixtures). Under conditions of a high contrast for the protein constituent the radius of gyration of the post-translocation-state ribosomes was found to be slightly greater than that of the pre-translocation-state ribosomes. Using the results of this study the conclusion can be drawn that translocation is accompanied by a spatial displacement of some parts of the ribosome with a magnitude of several ångström units.

Escherichia coli

[Oxygen consumption of the Baikal seal during unrestrained swimming in a pool and diving of different duration].

In 7 male Baikal seals (14-49 kg) spending 60-90% of the time under water, swimming unrestrained in the swimming pool, the average VO2 ranged from 8.6 +/- 7.01 ml (kg min)-1 for a 14-kg seal up to 4.8 +/- 2.22 ml (min kg)-1 (+ sigma) for adult 49-kg seal. A short forced diving (3-5 min) induced no change of the VO2 level because of stored oxygen. More prolonged diving (10-30 min) induced a 2-3-fold reduction of general VO2 and accumulation of a considerable O2-dept. During 3-5 min, 50-80% of this debt is covered in the 1st phase of recovery. The rest of the debt is covered during 10-40 min, depending on diving duration, body mass and, probably, oxidizing of metabolites of anaerobic exchange.

Animals

Structural study of translating 70 S ribosomes from Escherichia coli. I. Electron microscopy.

Translating 70 S ribosomes of Escherichia coli either in the pre-translocation or in the post-translocation state have been prepared by using the cell-free translation system in poly(U)-S-S-Sepharose columns [Methods Enzymol. (1979) 59, 382-398]. Electron microscopy study of the preparations has demonstrated that: (1) the mutual orientation of the ribosomal subunits in the translating ribosomes is the same as proposed by Lake for routine 30 S.50 S couples [J. Mol. Biol. (1976) 105, 111-130]; (2) the L7/L12 stalk of the 50 S subunit sticks out from the 70 S particle and does not join the 30 S subunit; (3) pre-translocation and post-translocation state ribosomes do not differ in mutual orientation of the subunits and in the position of the L7/L12 stalk, within the limits of electron microscopy resolution.

Cell-Free System

[Preparation of translating ribosomes by using a column with immobilized polyuridylic acid].

An improved method for preparation of translating ribosomes using columns with immobilized polyuridylic acid is described. A peculiarity of the method is that, first, optimal ratios of the components are used in the translation system for obtaining high yields of translating ribosomes. Second, purification of translating ribosomes from admixtures of non-translating particles is achieved by passing the buffer containing 5 mM MgCl2 and 250 mM NH4Cl through the column. The purity of the translating ribosomes is no less than 95%, the yield of active ribosomes is 5-20% of the initial amount of the ribosomes. Reagent expenditure is cut 15 to 20 times.

Methods