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V S Ivanova

Publications and source records attributed to V S Ivanova.

At least 19 recordsLinked to original sources

[Primary characterization of the properties and diagnostic value of the antigenic complex "fraction V" in a plague microbe].

The experimentally obtained antigenic complex isolated by extraction in the gradient surfactants from live and acetone-dried bacteria of the capsule-free vaccine strain EV76 of a plague microbe that had lost its ability to synthesize the diagnostic species-specific capsular antigen F1 was investigated. The antigenic complex fraction V (FV) was obtained after the fifth stage of extraction at a concentration of 1.28% of surfactants and after additional purification. The thermostable FV was found to consist mainly of protein. The protein having a molecular mass of about 43 kD predominates in the fraction. The latter is nontoxic for albino mice and antigenic. It forms a precipitate with commercial antiplague serum antibodies. FV antigenic sensitization of tanned sheep red blood cells gave rise to a diagnostic agent that specifically reacted with an antiplague serum rather than with heterologous sera against enterobacteria. The sera immunized with FV specifically reacted in the JDJFR with all the strains of the pathogen of plague irrespective of the temperature of their cultivation, including "fraction-free", which did not interact with a diagnosticum on F1. The animal sera immunized with capsule-free plague microbial strain reacted only with a FV-erythrocytic diagnosticum and they did not interact with F1 antigen-sensitized red blood cells. The erythrocytic FV diagnosticum was tested in ABNR with 130 typical and atypical plague microbial strains and with 133 strains of heterologous bacteria of different species of the family Enterobacteriaceae. The FV diagnosticum identified all the variants of a plague microbe, while the F1 diagnosticum revealed only its capsular variants. Among the heterologous bacteria, some strains of the closely related pathogen of pseudotuberculosis in those who were in the R form, rather than S form, positively reacted. The use of FV identified 2 groups of hybridomas obtained after immunization of albino mice with the capsule-free variant of a plague microbe. Some hybridomas reacted only with plague bacteria while others did with two above pathogens. The authors substantiate the expediency of using FV, its components, and obtained monoclonal plague pathogen antibodies to improve antiplague diagnosticums with an activity spectrum that exceeds that of the existing commercial F1 antigen-based diagnosticums. They also discuss the lines of further studies.

Animals↗

[Characteristics of plague pathogen mutants, differing by pigment sorption signs].

Wild-type strains of plague agent Yersinia pestis are characterized by a pigmentation phenotype (Pgm+), which includes several traits: an ability of cells to adsorb pigments (Hms+), an ability to produce siderophore yersiniabactin (Ybt+) and an ability to cause lethal infections in laboratory animals (Vir+) after subcutaneous injections. All these traits are encoded in the chromosomal pgm-locus, which gets rapidly lost due to deletion. One more trait related with the Pgm+ phenotype was detected in the present study, i.e. its siderophoric activity at 28 degrees C on the indication agar plates containing chrome azurol S (Sid+). After the four phenotypic characteristics of the Pgm+ phenotype were analyzed as well as after the four pgm-locus genes (hmsF, hmsR, irp2 and fyuA/psn) were detected by the method of hybridization and PCR, we compared 33 isogenous Pgm- mutants isolated from typical Y. pestis strain 923 by Hms-. The comparison showed that the mutants differed from each other according to the analyzed properties, which suggested that they were formed by different genetic mechanisms. Apart from the known mechanism of pgm-locus deletion, which causes an irreversible loss of Hms+, Ybt+ and Vir+ properties, two more mechanisms were detected. One of them is related with insertion damages to the pgm-locus genes, which also leads to the loss of the four traits but which can be reversed by the cultivation of cells at low temperature. The other mechanism is predetermined by unknown genetic processes ensuring the formation of mutants, which loose only their Hms+ properties and which can trigger its high-frequency reversion at 28 degrees C.

Base Sequence↗

[Study of PGM mutation mechanism in Yersinia pestis (plague pathogen) vaccine strain EV76].

Yersinia pestis vaccine strain EV76 is a mutant of the virulent strain which has lost the pigmentation phenotype (Pgm+). This phenotype includes three characteristics: it absorbs pigments from agar media (Hms+), produces a siderophore yersiniabactin (Ybt+), and causes a lethal disease after subcutaneous inoculation of laboratory animals (Vir+). These characteristics are lost simultaneously after high frequency spontaneous deletion of 10 kB fragment of chromosomal DNA, termed the pgm locus. We compared the pgm locus-associated genetic and phenotypical properties of the vaccine strain with those of a typical Pgm- deletion mutant of a virulent strain. The results indicate that Pgm- phenotype of the vaccine strain results not from the deletion of the pgm locus, but from the insertion inactivation of the genes located in this locus. In contrast to the deletion mutant, the vaccine strain carries sequences detected by hybridization and PCR, which are complementary to the pgm locus genes. Moreover, the vaccine strain differed from the deletion mutant by a low level of Hms+ expression, a slower rate of cell death under iron-chelated conditions at 37 degrees C, "residual virulence" upon subcutaneous inoculation, and capacity to form revertants which restore the characteristics of Pgm+ phenotype after cell growth at 12 degrees C.

Animals↗

Characterization of the stability and folding of H2A.Z chromatin particles: implications for transcriptional activation.

H2A.Z and H2A.1 nucleosome core particles and oligonucleosome arrays were obtained using recombinant versions of these histones and a native histone H2B/H3/H4 complement reconstituted onto appropriate DNA templates. Analysis of the reconstituted nucleosome core particles using native polyacrylamide gel electrophoresis and DNase I footprinting showed that H2A.Z nucleosome core particles were almost structurally indistinguishable from its H2A.1 or native chicken erythrocyte counterparts. While this result is in good agreement with the recently published crystallographic structure of the H2A.Z nucleosome core particle (Suto, R. K., Clarkson, M J., Tremethick, D. J., and Luger, K. (2000) Nat. Struct. Biol. 7, 1121-1124), the ionic strength dependence of the sedimentation coefficient of these particles exhibits a substantial destabilization, which is most likely the result of the histone H2A.Z-H2B dimer binding less tightly to the nucleosome. Analytical ultracentrifuge analysis of the H2A.Z 208-12, a DNA template consisting of 12 tandem repeats of a 208-base pair sequence derived from the sea urchin Lytechinus variegatus 5 S rRNA gene, reconstituted oligonucleosome complexes in the absence of histone H1 shows that their NaCl-dependent folding ability is significantly reduced. These results support the notion that the histone H2A.Z variant may play a chromatin-destabilizing role, which may be important for transcriptional activation.

Chromatin↗

DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139.

When mammalian cell cultures or mice are exposed to ionizing radiation in survivable or lethal amounts, novel mass components are found in the histone H2A region of two-dimensional gels. Collectively referred to as gamma, these components are formed in vivo by several procedures that introduce double-stranded breaks into DNA. gamma-Components, which appeared to be the only major novel components detected by mass or 32PO4 incorporation on acetic acid-urea-Triton X-100-acetic acid-urea-cetyltrimethylammonium bromide or SDS-acetic acid-urea-cetyltrimethylammonium bromide gels after exposure of cells to ionizing radiation, are shown to be histone H2AX species that have been phosphorylated specifically at serine 139. gamma-H2AX appears rapidly after exposure of cell cultures to ionizing radiation; half-maximal amounts are reached by 1 min and maximal amounts by 10 min. At the maximum, approximately 1% of the H2AX becomes gamma-phosphorylated per gray of ionizing radiation, a finding that indicates that 35 DNA double-stranded breaks, the number introduced by each gray into the 6 x 10(9) base pairs of a mammalian G1 genome, leads to the gamma-phosphorylation of H2AX distributed over 1% of the chromatin. Thus, about 0.03% of the chromatin appears to be involved per DNA double-stranded break. This value, which corresponds to about 2 x 10(6) base pairs of DNA per double-stranded break, indicates that large amounts of chromatin are involved with each DNA double-stranded break. Thus, gamma-H2AX formation is a rapid and sensitive cellular response to the presence of DNA double-stranded breaks, a response that may provide insight into higher order chromatin structures.

Amino Acid Sequence↗

Characterization of the human histone H2A.X gene. Comparison of its promoter with other H2A gene promoters.

The human gene for the replication-unlinked histone protein H2A.X is a naturally occurring chimera that contains a replication-unlinked promoter yet produces a stemloop mRNA characteristic of replication-linked histone genes. Consistent with the latter attribute, the H2A.X gene was found to lack introns. The promoter of the H2A.X gene was localized to a 120-base pair region upstream of the transcription start site, a region which included a TATA and two CCAAT sequence elements. The proximal of the two CCAAT elements was shown to be an important determinant of H2A.X gene promoter activity. In a comparative study with the CCAAT elements from the replication-linked H2A.1a gene and the replication-unlinked H2A.Z gene, the proximal CCAAT element of the H2A.X gene was found to bind nuclear factors also bound by CCAAT elements in the latter but not in the former. The specificity of the replication-unlinked H2A.X and H2A.Z gene promoters for CCAAT-binding transcription factors appeared to also reside in short homologous sequences about 10 base pairs away on either side of the CCAAT sequence.

Animals↗

Expression of a marked H2A histone protein in mammalian cells.

Histone protein sequences are highly conserved. In order to determine whether histones with sequences not found in nature would be tolerated in the chromatin of tissue culture cells, a gene for histone H2A.1a was altered by extending the protein coding region with eight amino acids, including three residues for methionine which are lacking in H2A.1a. Isolated clones of HeLa cells transfected with the gene construct were found to produce a novel protein which was resolved from other histone proteins on AUT-AUC two-dimensional gels. One clone, HeLa-B4, in which the novel protein named H2A.E accounted for about 10% of total H2A protein, was studied further. The linkage of H2A.E mRNA concentrations to the rates of DNA and protein synthesis was found to be the same as that of other replication-linked histone mRNA species. The stability of H2A.E in chromatin as well as the partitioning of nascent H2A.E protein between soluble and nuclear fractions was found to be indistinguishable from that of other histone species. This study shows that histone proteins with sequences other than the conserved sequences found in nature may be utilized in tissue culture cells.

Amino Acid Sequence↗

Chromosomal localization of the human histone H2A.X gene to 11q23.2-q23.3 by fluorescence in situ hybridization.

The human histone H2A.X gene is unusual in that its transcripts are alternatively processed to yield two species, one a 0.6-kb replication-linked histone mRNA and the other a 1.6-kb polyadenylated mRNA. The H2A.X gene has been localized by fluorescence in situ hybridization to chromosome 11q23.2-q23.3, away from the known clusters of human histone genes on chromosomes 1, 6, and 12. Assignment to chromosome 11 was substantiated by analysis of human-hamster somatic cell hybrid lines. As this work was being completed, an 89-bps sequence overlap was found between the downstream regions of the H2A.X gene and the recently sequenced hydroxymethylbilane (HMB)-synthase gene. The H2A.X and HMB-synthase genes have an unusual arrangement, being transcribed towards each other with their polyadenylation sites 330 bp apart. In addition the HMB-synthase gene contains constitutive and erythroid specific promoters. K562, an erythroid cell line, was found to contain a high concentration of the 1.6-kb polyadenylated H2A.X mRNA.

Animals↗

[Comparative study of the quality of domestic albumin solution and foreign analogs].

The quality and standards of albumin solutions (5, 10 and 20%) made in this country meet the requirements of national specification documentation. This, however, lacks some standards included into European Pharmacopoeia and mandatory for foreign manufacturers (Na and K ions, hemipigments, polymers, thermostability). The comparative tests of the albumin solutions made in Russia and abroad by conventional European standards showed that Russian solutions by some parameters are inferior to foreign samples. This urges improvement of the solution production technology as well as updating technical documents regulating the product quality.

Quality Control↗

[Results of hematological examinations of crew members of the space vehicles EO-2, EO-3 and EO-4].

The authors' opinion that adaptation inhibition of erythron functioning under weightlessness produces an unfavourable effect on the optimal physical state of a cosmonaut and his working capacity in the post-flight period has been confirmed by the analysis of certain hematological parameters studied in cosmonauts. The data obtained have necessitated investigation of threshold values of erythron functioning under weightlessness, and creation of artificial gravitation on board the piloted space ship to normalize erythropoiesis. Investigation of these problems of space hematology would be helpful in validation of the limits of human's long-term stay under weightlessness, and in the pre- and post-flight period management.

Adaptation, Physiological↗

[Regulation of erythropoiesis in patients with iron deficiency anemia].

Erythropoietin level in the blood plasma, iron metabolism, and some erythron parameters characterizing anemia expression, erythropoiesis effectiveness and processes of hemoglobin synthesis were studied in patients with iron-deficiency anemia (IDA). The results of the investigation helped the authors to establish that hormone production in IDA patients is under the control of a feedback mechanism that is functioning at a lower level as compared to other non-renal anemias. It has been suggested that optimum possible erythron functioning in IDA patients is achieved under these conditions.

Adolescent↗

[Various properties of RifR mutants of the plague agent].

The properties of the RifR mutants of four vaccinal and two natural strains of the plague bacillus were studied. The frequency of the mutants not growing on the complete nutrient medium after increasing the cultivation temperature to 37 degrees C averaged to 3.10(-1). The frequency of the loss of every of the three known autonomous plasmids or combinations of the 6-mD and 65-mD plasmids was n.10(-1). In the mutants of some strains with the preserved 47-mD plasmid sensitivity to the Ca2+ deficiency markedly lowered (but was not lost). The mutants of the majority of the strains produced much lower quantities of bacteriocin (pesticin I). The nutritional requirements, enzymatic activity, sensitivity to the diagnostic phage and the level of the production of fraction I, a specific antigen, in the bacillus did not change.

Bacteriocins↗

[Structure and antitumor activity of polysaccharides from the micelles of Aspergillus oryzae].

Three polysaccharide fractions active against mammary gland adenocarcinoma Ca-755 in mice but inactive against sarcoma C-180 were isolated from Aspergillus oryzae strain 5214. The main fraction, extracted from the mycelium by cold dilute alkali in the presence of sodium borohydride, was shown to be linear (1----3)-a-D-glucopyranan (pseudonigeran) according to 13C NMR spectroscopy, partial acid hydrolysis and periodate oxidation data.

Adenocarcinoma↗

[Erythropoietin in differential diagnosis of erythremia and secondary erythrocytosis].

The results of estimating blood plasma erythropoietic activity in patients with polycythemia of unclear genesis, in 95% of cases coincided with the clinical diagnosis proved during further follow up. These data have permitted recommendation of estimating blood plasma erythropoietic activity in patients to verify the diagnosis of polycythemic states.

Adult↗

[Screening of plasmids in museum strains of Yersinia pestis isolated from various natural foci].

92 strains of Yersinia pestis isolated from different natural foci and stored for 3-40 years in the museum of live cultures have been studied. The strains having three typical plasmids, their different combinations, plasmidless strains or the strains carrying nontypical plasmids with the molecular masses 9, 15, 55, 80, 90 and 150 Md were found. The old museum strains are proposed to be used as a source of plasmids for the genetical research. The current control of plasmid contents in the museum strains is suggested by the plasmid changes in course of storage.

Conjugation, Genetic↗

[The phagocytic activity of peritoneal macrophages in relation to Yersinia pestis with defective and complete Fra genes].

In vitro study of phagocytosis has shown that in guinea pigs fraction 1 is conducive to the ingestion of Y. pestis by macrophages, to survival and proliferation of Y. pestis cells in these macrophages, as well as to their specific transformation leading to their increased ingestive and bactericidal activity with respect to Y. pestis. In mice the role of fraction 1 in phagocytosis has proved to be less significant.

Animals↗

[Erythropoiesis inhibitor in the plasma of patients with hemopoietic dysplasias].

Altogether 21 patients were examined for an erythropoiesis inhibitor contained by the IgG fraction of the blood plasma. All these patients suffered from hemopoiesis depression. The inhibitor identified in the immunoglobulin fraction of the blood plasma of 40% of the patients manifested itself at the stage of erythroid precursors of bone marrow cells. The role of the humoral inhibitor in the development of erythropoietic disorders in these patients is under discussion.

Animals↗