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V V Safronov

Publications and source records attributed to V V Safronov.

At least 19 recordsLinked to original sources

[Loss and stabilization of aminopterin resistance in murine cell lines].

Mouse L-cell lines (B-82, tk-) were obtained using the stepwise selection method, their aminopterin (AP) resistance being 10(3)-5 X 10(4) times higher than that of parental cells. This resistance increase results from dihydrofolate reductase (DHFR) gene amplification which was determined from the 15-120-fold rise of the enzyme activity and with the cytogenetical techniques. The development and loss of AP resistance have been studied and karyological analysis of the lines obtained carried out. Two types of karyological changes were found in stable DM and HSR cells which correspond to extrachromosomal and intrachromosomal forms of the amplified material organization. Localization of the DHFR gene in HSR was proved using the in situ hybridization technique. Extrachromosomal localization of the amplified genes in DM providing unstable AP resistance is dominant at the early stages of the development of resistance and for a long time. It was demonstrated that DM and HSR can exist in one cell during the prolonged period. DHFR gene copy number in such cells is regulated by a change in the DM number, whereas the HSR size and localization are highly stable. HSR covers 1.7-1.9% of the genome length and 38-40% of the marker chromosome length. The genes localized in HSR provide stable AP resistance. Evidence on some intermediate, relative stabilization of the resistance has been obtained. This stabilization is mediated by temporary integration of DHFR copies into other chromosomal sites, in addition to HSR.

Aminopterin

[Distribution of spontaneous and 8-methoxypsoralen-induced sister chromatid exchanges along the length of the 1st chromosome of Chinese hamster cells].

Distributions of spontaneous, induced by monoadducts and induced by crosslinks sister chromatid exchanges (SCE) along the first chromosome of Chinese hamster cells are nonrandom. All experimental distributions have low frequency of SCE in centromeric and telomeric regions. It can be explained by specific structural organization of the chromosome. However, there are some differences between experimental distributions. Distribution of SCE induced by crosslinks differs from that of spontaneous SCE. Distribution of SCE induced by monoadducts, unlike other distributions, has an increased frequency of exchanges in the q11 region. This region contains several narrow closely disposed G+ bands. It is possible that monoadducts lead to increasing SCE frequency on G+-G- junctions. Distribution of SCE induced by crosslinks resembles random distribution, except centromeric and telomeric regions. These results lead to conclusion that the mechanisms of formation of spontaneous, induced by monoadducts and induced by crosslinks SCE differ from each other.

Animals

[Biochemical and ultrastructural analyses of the synaptonemal complex in mammalian spermatocytes].

In order to study the molecular organization of synaptonemal complex (SC), a preparative method for isolation of relatively purified SC from rat, mouse and hamster testes was elaborated which involves isolation of SC-containing (pachytene) nuclei, their lysis, DNAase digestion of DNA and fractionation of nuclear elements by the discontinuous sucrose density gradient centrifugation. Electron microscopy revealed a rather good preservation of the SC structure after the isolation procedure. Effects of the dissociating agents on the SC structural integrity were studied. It has been demonstrated that the treatment with 2M NaCl, Triton X-100, sodium deoxycholate, 6M urea, and with a buffer containing 2% SDS and 5% mercaptoetthanol does not lead to a complete SC dissociation, though it results in some structural chanes. Possible reasons of the high resistance of SC to dissociating treatment are discussed.

Animals

[Autoradiographic analysis of molecular mechanisms of differential staining of sister chromatids].

The significance of procedures involved in the technique of differential staining of sister chromatids is estimated by quantitative autoradiography. Photolysis of bromodeoxyuridine-substituted DNA is shown to be the main process determining the quality of differential staining under certain experimental conditions. On the basis of modes provoking DNA degradation and washing DNA fragments off the chromosomes, a simplified method including only irradiation of chromosomes under a mercury lamp and treatment with a sodium citrate solution is proposed.

Animals

[Duration of meiotic prophase stages during oogenesis in the newt Triturus cristatus cristatus].

The duration of the early stages of meiotic prophase was determined in the oogenesis of T. cristatus cristatus by means of autoradiography. The oocytes were being investigated during 39 days from the moment of 3H-thymidine injection. It was shown that preleptotene lasts 1--2, leptotene ca. 4, zygotene 5 and pachytene 26 days. When studying the preparations obtained 1 day after the injection of 3H-thymidine, the silver grains were found to be localized over the nuclei at all stages of meiotic prophase; this suggests the amplification of rDNA which begins in leptotene-zygotene and ends in early diplotene.

Animals