Search PubMed⌕ Search

Biomedical subjects

A Viron

Publications and source records attributed to A Viron.

31 records · Page 2Linked to original sources
PubMed1984

Structural organization of rat hepatocyte chromatin as visualized in thin frozen sections selectively stained for DNA.

We studied the structure of rat hepatocyte chromatin in situ using thin frozen sections selectively stained for DNA after aldehyde fixation. Our results indicate that intranucleolar chromatin is arranged into three different organization levels, confirming the observations on Epon-embedded chromatin. These are: completely extended DNA filaments, with a thickness of approximately 3 nm, clustered in loose, roundish agglomerates, very long fibers with a thickness ranging from 15 to 35 nm and compact chromatin clumps. Both the fibers and the chromatin clumps frequently appeared to be composed of nucleosome-like particles. In the extranucleolar chromatin, agglomerates of extended DNA filaments and long fibers were never visualized. In contrast to data from Epon-embedded chromatin, we noticed that in frozen sections neither the nucleolar nor the extranucleolar compact chromatin appear to be organized into discrete, 20 to 30 nm superordered fibers.

Animals↗
PubMed1983

Preferential nucleolar localization of Cis-DDP in human fibroblasts.

The use of complex platinum salts (Cis-DDP) in antitumoral therapy is well known. However the mode of action of this salt remains unknown. The present study was performed with human transformed cells in culture (WI-98-VAD), grown in the presence of Cis-DDP. The intracellular localization of platinum was studied by associating the technique of microanalysis by electronic probe (analysis by wavelength dispersion: W. D. S.) and that of fine tissue sections at low temperature: ultracryotomy. The platinum was found to be concentrated mainly in the nucleolar region.

Cell Nucleolus↗
PubMed1981

Effect of quinacrine on nuclear structure and RNA synthesis in cultured rat hepatocytes.

The effects of quinacrine, an antimetabolite which intercalates into DNA, on the ultrastructure of interphase nuclei and on RNA turnover were studied in primary cultures of rat hepatocytes. Procedures included ultrastructural cytochemical staining for ribonucleoprotein and DNA, autoradiography, and measurement of labeled uridine uptake and incorporation. Addition to the culture medium of a nontoxic dose (10 microM for 30 min) reduces the net accumulation of labeled uridine in RNA. This involves first heterogeneous RNA and then ribosomal RNA since their structural precursors, interchromatin fibrils and nucleolar fibrils, respectively, diminish in that order. Intranucleolar chromatin retracts, and perinucleolar chromatin becomes unusually condensed. A toxic dose (50 microM for 30 min) produces greater inhibition of tritiated uridine incorporation in RNA. This precedes and is not due to a drop in uridine uptake into the cells. Toxic doses produce unusually large clusters of interchromatin granules which are embedded in an unusual dense material which stains positively for ribonucleoprotein. Three regions of the chromatin are altered. (a) Perinuclear condensed chromatin retracts from the nuclear envelope, remaining attached by short DNA-containing bridges. (b) The normally dispersed nucleoplasmic chromatin condenses into a stainable network which retracts centrifugally. (c) Perinucleolar chromatin becomes a network of small highly condensed masses or bands interconnected by fibrils which are either decondensed or stretched. These alterations in chromatin structure probably form the basis of quinacrine-impaired nuclear metabolism.

Animals↗
PubMed1979

[Monolayer culture of adult mouse hepatic cells secreting albumin and alpha fetoprotein].

Adult mouse liver cells obtained by enzymatic dispersion were maintained in primary cultures for up to 4 weeks. They retained some of the typical morphological and ultrastructural characteristics of hepatocytes. After 2 days of culture, structures similar to bile canaliculi were found and after 6 days clusters of small proliferating cells were noticed in the gaps of the monolayer formed by large well-spread hepatocytes. Almost all 3-day cultures began to secrete alphafetoprotein for about 2 weeks, whereas albumin was secreted throughout the period of culture.

Albumins↗
PubMed1979

[Use of a cryoultramicrotomy method for the purpose of studying chlamydial ultrastructure].

The method of cryoultramicrotomy was adapted for the study of the ultrastructure of HeLa and McCoy cells in monolayer cultures infected with Chlamydia, obligatory intracellular procaryotic parasites, the causative agents of ornithosis (strain Loth) and paratrachoma (strain LB 1). The cryosections were obtained by the fixation of the monolayer with 2.5% glutaraldehyde, by the gradual infiltration of precipitated cells with sucrose (0.6--1.2--1.8--2.3 M) prior to freezing in liquid nitrogen, and by the treatment of sections with 1% aqueous methyl cellulose solution before drying. This method ensured good preservation of both Chlamydia, in intracytoplasmic inclusions and host cells, as well as regular reproducibility of the results. Ultrathin sections showed a considerable polymorphism in the vegetative forms of Chlamydia, which was probably due to the structure of their cell walls. Chlamydia, were found to form small vesicle-like structures in the cavities of inclusions. The cell walls and granules inside the elementary bodies of the causative agent of ornithosis were stained with the use of phosphotungstic acid--HCl, pH 0.5.

Chlamydia↗
PubMed1978

Morphological and cytochemical study of Chlamydia with EDTA regressive technique and Gautier staining in ultrathin frozen sections of infected cell cultures: a comparison with embedded material.

The cryo-ultramicrotomy technique was applied to study the ultrastructure of Chlamydia using two strains: one of C. psittaci and one of C. trachomatis. It clearly appeared that in both strains reticulate bodies show a high degree of plasticity, contrasting with the rigid spherical appearance of elementary bodies. Ultrastructural cytochemical study shows DNA fibrils dispersed throughout the cytoplasm in reticulate bodies whereas DNA is condensed in a nucleoid in elementary and intermediate bodies. The EDTA regressive technique reveals ribonucleoproteins in reticulate and elementary bodies of both studied strains.

Animals↗
PubMed1977

Diaminobenzidine cytochemistry in cryo-ultramicrotomy for the detection of peroxidases.

Rat erythrocytes and lymphoid cells were fixed by glutaraldehyde and encapsulated in bovine serum albumin plus rabbit IgG globulins for cryo-ultramicrotomy. A technical procedure is described by which endogenous peroxidases of erythrocytes in ultrathin frozen sections were detected by hydrogen peroxide and diaminobenzidine as hydrogen donor. Modifications of this classical cytochemical procedure proved also useful in cryoultramicrotomy for immunoperoxidase labeling of antigenic determinants in rabbit IgG globulins which have been crosslinked within the supporting matrix of cells.

3,3'-Diaminobenzidine↗
PubMed1971

Improved techniques for the preparation of ultrathin frozen sections.

Ultrathin frozen sections of biological tissues for electron microscopy provide certain advantages in cytochemical studies in which the penetration of cells by large molecules is necessary and in morphological studies of cellular constituents which are dissolved by the reagents employed in routine plastic embedding. The recent introduction of several types of commercially available cryo-ultramicrotomes makes it possible for many laboratories to employ this valuable tool. This paper summarizes recent improvements in the methods developed in this laboratory for preparing ultrathin frozen sections and reviews some of the inherent problems involved in their use. These procedures may serve as a baseline for other investigators who can then modify or adapt them for their specific purposes.

Adrenal Glands↗