Biomedical subjects

E Stephanova

Publications and source records attributed to E Stephanova.

8 recordsLinked to original sources

PubMed2005

Volatile anaesthetic halothane causes DNA damage in A549 lung cells.

The present study was performed to elucidate the extent of damage and the ability of lung epithelial cells to recover or to undergo apoptosis after in vitro treatment with the volatile anaesthetic halothane. The results obtained from the comet assay clearly show that halothane, applied at 3.0mM concentration, causes DNA and cell damage. Cells exhibited nuclear fragmentation and budding early after treatment and these events gradually increased during the next few days. The presence of a large number of mini-comets after single cell gel electrophoresis was found to represent apoptotic bodies with fragmented DNA. Our results demonstrate apoptosis-like changes after in vitro exposure of A549 cells to the volatile anaesthetic halothane. The majority of the affected cells did not recover and were directed to cell death.

Anesthetics, Inhalation↗

PubMed1997

Rat chromosome 1: regional localization of seven genes (Slc9a3, Srd5a1, Esr, Tcp1, Grik5, Tnnt3, Jak2) and anchoring of the genetic linkage map to the cytogenetic map.

Seven genes were regionally localized on rat Chromosome (Chr) 1, from 1p11 to 1q42, and two of these genes were also included in a linkage map. This mapping work integrates the genetic linkage map and the cytogenetic map, and allows us to orient the linkage map with respect to the centromere, and to deduce the approximate position of the centromere in the linkage map. These mapping data also indicate that the Slc9a3 gene, encoding the Na+/H+ exchanger 3, is an unlikely candidate for the blood pressure loci assigned to rat Chr 1. These new localizations expand comparative mapping between rat Chr 1 and mouse or human chromosomes.

Animals↗

PubMed1996

Localization of nucleic acids in hepatocyte nucleoli of rats upon D-galactosamine-induced block of transcription.

The precise localization of DNA and RNA within rat hepatocyte nucleoli during the process of D-galactosamine-induced nucleolar segregation has been studied by using sensitive methods for their detection: osmium-ammine staining and terminal deoxynucleotidyl transferase reaction for DNA, and immunoelectron microscopy with anti-RNA antibodies, RNase-gold, and autoradiography with tritiated orotic acid for RNA. The blocking of transcription was followed by the disappearance of intranucleolar condensed chromatin. Agglomerates of thin extended DNA filaments were found to change their location to the nucleolar periphery and to coalesce with each other. At the last stage of nucleolar segregation they were concentrated at the pole of the nucleolar fibrillar remnant while the rest of the nucleolus did not contain any DNA. No DNA was found in the dense fibrillar component of both intact and treated hepatocyte nucleoli. During the process of nucleolar segregation the bulk of the nucleolar RNA was found within the so-called spherical bodies. This RNA appeared to be synthesized shortly before or even after drug administration. The results obtained are in agreement with the hypothesis that the fibrillar centers are the site of nucleolar transcription. They also show that uncompleted molecules of pre-rRNA whose synthesis has been blocked are segregated from the rest of nucleolar RNA species into the spherical bodies.

Animals↗

PubMed1996

The rat genes encoding the pancreatitis-associated proteins I, II and III (Pap1, Pap2, Pap3), and the lithostathin/pancreatic stone protein/regeneration protein (Reg) colocalize at 4q33-->q34.

Using fluorescence in situ hybridization, we determined that the three rat PAP genes, and the related REG gene map in the same chromosomes region, namely 4q33-->q34. This rat chromosome region is thus homologous to the human 2p12 region, which also contains the PAP gene, the REG1A gene, and a REG-related gene (REGL).

Acute-Phase Proteins↗

PubMed1993

Binding of sequences from the 5'- and 3'-nontranscribed spacers of the rat rDNA locus to the nucleolar matrix.

Nucleolar matrix structures were obtained under different extraction conditions from highly purified isolated nucleoli. Their ultrastructural appearance, protein composition and capacity to bind rDNA preferentially were studied in a model binding system. A region spanning approximately 25 kb in the rat ribosomal gene locus was screened for DNA sites capable of specifically interacting with the proteins of the nucleolar matrix (MARs). Two such sites were identified: one is located on an EcoRV-KpnI fragment in the 5'-nontranscribed spacer region, between two repetitive elements and close to the transcription initiation site; the other MAR is on a PvuII-BamHI fragment located in the 3'-nontranscribed region, encompassing an element 85% homologous to a B2-sequence. The two MARs are located in regions rich in polypyrimidine/polypurine tracks and contain a few elements homologous to the consensus sequence for topoisomerase II. This indicates that the "attachment sites" for the ribosomal genes belong to the same class of sequences as the MARs attaching the chromosomal DNA to the nuclear matrix.

Animals↗

PubMed1989

Histone variants in mouse centromeric chromatin.

Highly purified centromeric heterochromatin was isolated from mouse liver nuclei and the pattern of core histone variants was analyzed. In comparison with total chromatin, the centromeric heterochromatin of young animals was characterized by (1) enrichment in the replication-dependent variants H2A1, H2B2 and H3(2), (2) reduced amount of the minor variant H2Az and (3) absence of ubiquitinated molecules of H2A. This specific variant pattern changed upon ageing as a result of accumulation of replacement variants so that in adult animals both chromatin preparations exhibited similar pattern for H2A and H2B, while the difference in the profile of H3 variants was preserved.

Aging↗

PubMed1988

Mouse centromeric heterochromatin: isolation and some characteristics.

A method is suggested for isolation of highly purified mouse centromeric heterochromatin. Treatment of mouse liver nuclei with decreasing concentrations of Ca2+ resulted in the gradual unraveling of chromatin in the nucleus and at 0.1 mM Ca2+ electron microscopy revealed several dense particles per nucleus, surrounded by decondensed chromatin. These particles, assumed to represent centromere regions of interphase chromosomes by in situ hybridization with radioactive mouse satellite DNA and by differential staining for centromere heterochromatin, were isolated in preparative amounts and their DNA and protein composition was analyzed. The preparation represented practically pure mouse centromere heterochromatin, since more than 90% of its DNA was satellite DNA.

Animals↗

PubMed1983

Trout sperm chromatin. I. Biochemical and immunological study of the protein composition.

Compact sperm chromatin was obtained from mature trout sperm nuclei resistant to sonication and detergent treatments. 0.5 to 2 M NaCl caused a gradual decondensation of this chromatin and the dependence of the percentage of dissociated proteins on the salt concentration indicated cooperativity of the dissociation process. Urea alone was insufficient to decondense the nuclei. The only proteins dissociated from the sperm nuclei by NaCl alone or combined with urea were protamines. Besides protamines, tightly bound nonprotamine proteins resisting high salt-urea extraction were detected in the sperm nucleus. Part of them could be solubilized by 1% sodium dodecyl sulphate (SDS) and displayed the characteristics of the core histones: they were soluble in 0.25 N H2SO4, their electrophoretic mobilities were similar to those of trout liver core histones, and they shared common antigenic determinants with the latter. The rest of the tightly bound proteins resisted 1% SDS treatment and could be obtained after an extensive digestion of DNA with DNase I. These were nonhistone proteins similar in mobility to the protein triplet characteristic of the lamina-pore complex and an additional high molecular weight protein.

Animals↗