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Biomedical subjects

V I Kiselev

Publications and source records attributed to V I Kiselev.

At least 19 recordsLinked to original sources

Developing additional resources.

We briefly outline existing information about several cohorts in the Southern Urals, Semipalatinsk and the Altai regions, in addition to those discussed in the companion papers in this issue of Radiation and Environmental Biophysics. These include: (a) the East-Urals Radiation Trace (EURT) cohort of individuals (exposed to fallout from the September 1957 explosion of a storage tank containing liquid radioactive waste from the Mayak Production Association) as well as their offspring, (b) the cohort of exposed parents (i.e. workers of the Mayak facility) and their children, having been established with the aim of examining reproductive health, and (c) several additional cohorts in the Altai region and in Semipalatinsk, where investment of additional resources would greatly facilitate the progress of ongoing studies. Furthermore, we include a brief description of the Russian Human Radiobiology Tissue Repository, which has been established in the city of Ozyorsk and is in the process of making an inventory of autopsied tissues from 700 deceased Mayak workers and of collecting and storing donations of blood and tumor tissues from the members of the Mayak workers cohort currently residing in the city.

Cohort Studies↗

A registry for exposure and population health in the Altai region affected by fallout from the Semipalatinsk nuclear test site.

A registry of the rural population in the Altai region exposed to fallout from nuclear tests at the Semipalatinsk test site (STS) was established more than four decades after the first Soviet nuclear explosion on August 29, 1949. Information about individuals living in an exposed and a control area was collected using all available local sources, such as kolkhoz documentation, school registries, medical treatment records and interviews with residents. As a result, a database comprising an exposed group of 39 179 individuals from 53 Altai region villages, 6769 external and 3303 internal controls was compiled. For several settlements, effective dose estimates reached the level of 1.5 Sv, while the average effective dose estimate in the exposed group was 340 mSv. Dosimetric data, vital status information and health records gathered at rayon and village medical facilities are held in the registry. Cause-of-death information for deceased residents is obtained from death registration forms archived at the Altai region vital statistics office. At present, a follow-up of approximately 40% of the population exposed in 1949 has been done. More will be added by searching for migrants to the larger towns of the Altai region, i.e. Barnaul, Rubtsovsk and Biisk. In order to assess the influence of radiation exposure, analytical studies with a case-control design for stomach and lung cancer are currently being prepared. The number of known cases is sufficient to detect an odds ratio of 1.5 at the 95% confidence level. Epidemiological studies in populations affected by fallout from STS may be equally important to the atomic bomb survivors' study for the direct quantification of radiation effects. The range of exposure rates experienced will extend the acute high-dose-rate findings from Hiroshima/Nagasaki towards acute and protracted lower exposures, which are more relevant for radiation protection issues.

Environmental Exposure↗

[Cloning the Staphylococcus aureus enterotoxin B gene, obtained by polymerase chain reaction, and its expression in Escherichia coli cells].

To determine the Staphylococcus aureus enterotoxigenicity, we have developed an approach based on polymerase chain reaction (PCR). Using this method several S. aureus strains have been screened for the presence of the enterotoxin B gene. A DNA fragment of the selected strain (FRI 722H) containing enterotoxin B gene has been obtained by the PCR method and cloned in the pUC19 vector. It is shown that enterotoxin B with the leader peptide forms insoluble complexes in E. coli cells, whereas the mature toxin is present in cytoplasmic fraction in a soluble form. The recombinant toxin made up for 1.7% of the total cellular protein in E. coli JM 109 cells.

Base Sequence↗

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↗

Ca(2+)-dependent phosphorylation of synapsin I as a possible regulatory mechanism of neurosecretion.

Phosphorylation of homogeneous synapsin I isolated from human brain by Ca2+, phospholipid-dependent protein kinase (protein kinase C) from the same source was studied. The inhibitory effect of calmodulin on this process was demonstrated. The kinetics of activation of synapsin I phosphorylation by acidic phospholipids, phosphatidylserine and phosphatidylinositol, in the absence and presence of phosphatidylinositol-4,5-bisphosphate and diacylglycerol was compared. The proteolytic effect of degradation of the synapsin I molecule phosphorylated by Ca2+, calmodulin-dependent protein kinase II was revealed. No proteolysis of synapsin phosphorylated under similar conditions either by protein kinase C or cAMP-dependent protein kinase was detected. In view of the process specificity, the physiological significance of the observed effect is suggested. The inter-relationship between two ways of neurosecretion regulation is discussed: an earlier known, conventional way, mediated by synapsin I phosphorylation by Ca2+, calmodulin-dependent protein kinase II, and another one, mediated by synapsin I phosphorylation by protein kinase C. The modulating role of polyphosphoinositides in the PK C-dependent way of regulation is considered.

Brain↗

Hydrophobized antiviral antibodies and antisense oligonucleotides.

A method of suppressing virus reproduction in cells has been proposed. The approach consists of affecting the cells with antiviral antibodies artificially hydrophobized with fatty acid residues. Reproduction of influenza viruses in MDCK cells and respiratory-synticial virus in HeLa cells was used as a model to demonstrate that poly- and monoclonal antibodies, modified by 1 or 2 stearic acid residues, are potent, unlike the non-modified antibodies, at inhibiting viral reproduction. The observed phenomenon is apparently due to penetration of hydrophobized antibodies into the cells. Thus, in particular, considerable antiviral activity is exhibited by monoclonal antibodies against NP-protein of influenza virus, which is an antigen accessible to antibodies only inside the infected cells. Hydrophobized antibodies do not affect the kinetics of viral protein synthesis; they block the virus withdrawal from the cells, probably by interfering with the assembling and budding of virus particles. To enhance penetration of oligonucleotides ("oligos") into cells, chemical modification of the former at the 5'-end phosphate group by fatty radicals has been suggested. The undecanol-modified oligo namely an oligo complementary to the protein binding sites located at the influenza virus polymerases encoding RNA, was synthesized using a DNA-synthesator. The above modified oligo effectively suppressed the influenza A/PR8/34 virus reproduction and inhibited synthesis of the virus-specific proteins in MDCK cells. The non-modified antisense oligo and the modified nonsense oligo did not affect the virus development under the same conditions.

Animals↗

[Neuroendocrine regulation under maximal and subjectively optimal physical loads].

The responses of neuroendocrine system to maximum and subjective-optimum physical loading were found to be similar in young men and characterized by activation of adrenocortical, somatotropic and thyroid functions, particularly under maximum loading. The subjective-optimum loading was dosed out by the subjects themselves, differentiated individually and averaged 62% of the maximum oxygen consumption. The parameters of subjective-optimum physical loading were close to the threshold level necessary to stimulate neuroendocrine system and ensure an increase of somatotropine, cortisole, thyroxine and aldosterone concentration in the blood.

Adolescent↗

A new class of antivirals: antisense oligonucleotides combined with a hydrophobic substituent effectively inhibit influenza virus reproduction and synthesis of virus-specific proteins in MDCK cells.

To enhance the penetration of oligonucleotide ('oligo') into cells, the oligo was combined with the hydrophobic undecyl residue. Using the 'DNA-synthesator', we synthesized oligo, complementary to the loop-forming site of the RNA, encoding polymerase 3 of the influenza virus (type A), and combined it with the undecyl residue added to the 5' terminal phosphate group. It was found that the modified oligo effectively suppresses the influenza A/PR8/34 (H1N1) virus reproduction and inhibits the synthesis of virus-specific proteins in MDCK cells. Under the same conditions, the non-modified antisense oligo and modified nonsense oligo did not affect the virus development.

Animals↗

Calmodulin-dependent enzymes as a target of staphylococcal enterotoxin A.

The response of lymphoid and nerve cells to the action of SEA has been investigated. It has been established that the toxin acts as a mitogen with respect to resting cells and suppresses the DNA biosynthesis in proliferating cells. Interaction of SEA with the systems of second messengers in lymphoblastoid cells has been studied. The results obtained suggest a mechanism of the antiproliferative action of SEA on these cells. Studies on the structural organization of the toxin molecule have revealed that the latter contains a polypeptide (BacM) capable of activating calmodulin-dependent enzymes both in the presence and absence of Ca2+. These findings permit us to assume that the cytostatic effect of SEA is conditioned by the formation of BacM and phosphorylation of elongation factor 2.

Adenylyl Cyclases↗

Amplification of DNA sequences of Epstein-Barr and human immunodeficiency viruses using DNA-polymerase from Thermus thermophilus.

Using thermophilic DNA-polymerase from Thermus thermophilus we have amplified by polymerase chain reaction (PCR) specific DNA sequences of Epstein-Barr virus (EBV) and human immunodeficiency virus (HIV). DNA-polymerase from Thermus thermophilus (molecular mass of 80-86 kDa) differs in its physico-chemical properties from DNA-polymerase from Thermus aquaticus (molecular mass of 62-68 kDa). To amplify the specific EBV DNA sequence, oligonucleotide primers for the virus replicon region (oriP region) were used. As a result of amplification, a specific 405-bp DNA fragment was produced.

Base Sequence↗

[Expression of Staphylococcus aureus enterotoxin A gene in heterologous systems].

The genomic library of Staphylococcus aureus genes on the plasmid vector pSL5 has been constructed. The library contains a 2.5 kb HindIII DNA fragment including the gene for enterotoxin A. The entA gene on the high copy number plasmids in the Escherichia coli cells deficient in proteolysis determines the synthesis of enterotoxin A in the amounts comparable to the ones in the parent strain Staphylococcus aureus FRI 722(H).

Bacillus subtilis↗

Effective inhibition of viral reproduction by hydrophobised antiviral antibodies.

A method is proposed for the inhibition of viral reproduction in cells by means of fatty-acylated antiviral antibodies which, in contrast to the unmodified antibodies, have the ability to enter the cells. The potential of this technique is demonstrated in experiments involving inhibition of the reproduction of various strains of influenza virus and respiratory syncytial virus.

Acylation↗