Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Cell Fractionation”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 145 records · Page 8Linked to original sources
PubMed1978

Studies on secretory glycoproteins in the rat exocrine pancreas. III. Intracellular transport of fucose-labeled proteins as studied by cell fractionation.

The transcellular movement of fucosylated glycoproteins has been studied in vitro using rat pancreatic lobules and cell fractionation procedures, and has been compared with the well established pathway of secretory proteins. Using tritiated leucine as pulse label for the latter, their translocation from the rough endoplasmatic reticulum into the Golgi complex and finally into zymogen granules could be followed. In the case of glycoproteins, 14C-fucose was incorporated mainly into the smooth microsomal fraction (representative of the Golgi complex) and only one third of this specific activity was transported into the zymogen granule fraction. A detailed analysis of this fraction after separation of the content of zymogen granules from their membranes revealed a predominant labeling of membrane glycoproteins by 14C-fucose. In comparison, leucine-labeled bulk proteins were found almost exclusively in the zymogen granule content fraction, with little radioactivity in the membrane fraction. The data indicate a concomitant synthesis of fucosylated glycoproteins destined in part for the zymogen granule membrane and to a greater amount associated with the smooth microsomal fraction. The results are discussed in the light of recent findings indicating that about 40% of the proteins in the zymogen granule membrane are made up of one major glycoprotein which could be involved in the mechanism of exocytosis.

Amylases↗
PubMed1976

Some biological properties of mouse spleen cells fractionated by the adherence of Sephadex G 25 and glass bead columns.

The possibilities of separation of haemopoietic cells from lymphocytes capable of eliciting the graft-versus-host reaction through column chromatography were investigated. Strain-A mouse spleen cells were fractionated into the adherent and non-adherent fraction on Sephadex G-25, glass bead columns and glass beads coated with antibody against mouse globulin. Increased numbers of cells forming haemopoietic colonies were found in the cell fraction which did not adhere to the antibody-coated glass beads and in cells reversibly adhering to glass beads. No significant decrease in local graft-versus-host reaction was found in any fraction obtained, and the prolonged survival of irradiated semiallogeneic recipients was observed in both fractions obtained on Sephadex G-25 columns.

Animals↗
PubMed1994

Transferrin binding capacity as a marker of differentiation and maturation of rat erythroid cells fractionated by counter current distribution in aqueous polymer two-phase systems.

Rat bone marrow cell populations, containing different proportions of erythroid cells, have been fractionated by counter-current distribution in the non-charge-sensitive dextran/polyethyleneglycol two-phase systems on the basis of hydrophobic cell surface properties. Cell fractions with a low distribution coefficient, which contain non-erythroid cells and early erythoblasts, showed a low transferrin binding capacity and a low haemoglobin/cell ratio whereas cell fractions with a high distribution coefficient, which contain intermediate-late erythroblasts and mature red cells, showed an elevated transferrin binding capacity and the highest haemoglobin/cell ratio. These results support transferrin binding capacity as a good marker parameter for the erythroid bone marrow cell differentiation and maturation processes.

Animals↗
PubMed1975

Isolation and subfractionation on ficoll gradients of adult rat hepatocytes. Size, morphology, and biochemical characteristics of cell fractions.

The recirculating perfusion of adult rat liver with a Ca-++-free Hanks' solution produces a release of the adhesiveness of cells and a cleaving of the desmosomes. The addition of collagenase and hyaluronidase to the perfusion medium leads to complete dissociation of the liver tissue into a mixture of isolated cells and cell cords in which the hepatocytes remain connected with specific junctional differentiations, namely the gap and tight junctions. Individual cells are released by submitting the suspension of cell trabeculae to a gentle rolling. The gap junctions are ruptured at least in one of the two adjacent cells and remain generally attached to the other cell taking with them a small portion of cytoplasm. This technique of isolation of hepatocytes yields about 60-65% of the parenchymal cells contained in a liver; endothelial cells and other cells of the connective tissue are not recovered. The ultrastructural preservation of the isolated hepatocytes is excellent and the glucose-6-phosphatase activity, confined to the endoplasmic reticulum, appears unaltered in most cells. Protein, DNA and RNA recovery in the preparations of isolated hepatocytes is satisfactory, amounting to 70% of that found in liver homogenate; glycogen, the most labile component examined, is partly lost or degraded during the manipulations. Cell diameters measured by different methods confirm the preservation of the original volume of the in situ hepatocytes and the presence of more than one type of parenchymal cell. By submitting this heterogeneous cell population to an isopycnic density gradient centrifugation, two types of hepatocytes can be distinguished: the light hepatocytes, with a mean diameter of 20.5 mum and a mean density of 1.10, are characterized by an extended smooth-walled endoplasmic reticulum entrapping dispersed alpha-glycogen particles; the heavy hepatocytes, with a mean diameter of 19.0 mum and a mean density of 1.14, present a relatively reduced compartment of smooth endoplasmic reticulum, but large accumulations of glycogen. It is suggested that the cell fraction of low density is enriched in centrolobular cells and the high density fraction in perilobular hepatocytes.

Animals↗
PubMed1995

Detection of hepatitis C virus RNA in the cell fraction of saliva before and after oral surgery.

The presence of hepatitis C virus (HCV) RNA in serum, whole saliva, and saliva from the submaxillary glands was investigated before and after oral surgery. The presence of HCV RNA (positive and negative-strand RNA) was determined in serum and saliva by a nested polymerase chain reaction in 26 anti-HCV positive patients, of whom 11 were coinfected with human immunodeficiency virus-1. Oral surgery was carried out on five occasions on four of the patients. HCV RNA was detected in the sera of 23 of 26 (88%) patients, and in the saliva of 4 of the 23 (17%) of the viremic patients. In all four cases, HCV RNA was detected only in the cell fraction derived from centrifugation of whole saliva. Negative-stranded HCV RNA was not detected. At one of five occasions of oral surgery, HCV RNA was detected in saliva sampled immediately after surgery, but not before or 24 hours after surgery. The results suggest that HCV is present in saliva in less than 25% of HCV viremic persons. The presence of the virus in saliva is restricted to the cell fraction. Thus, saliva may serve as a possible, but low, nonparenteral transmission route of HCV. Contamination of saliva by blood during and after oral surgery may result in an increased risk of viral exposure. Except for trauma caused by sharp instruments during surgery, this might contribute to the higher HCV seropositivity found among dentists.

Dentists↗
PubMed1993

Identification by mass spectrometry of CMP-NANA in diffusible material released from high M(r) blood cell fractions that confers serum resistance on gonococci.

In previous work, a low M(r) component from human blood which converts serum-sensitive gonococci to resistance was shown to be indistinguishable from cytidine 5'-monophospho-N-acetyl neuraminic acid (CMP-NANA) by seven criteria. However, the presence of CMP-NANA was not proved by physicochemical methods. Purified, high M(r) fractions from human blood cells, which confer serum resistance on gonococci and enhance the transfer of sialyl groups from CMP-NANA to lipopolysaccharide (LPS) by a sialyltransferase in gonococcal extracts, were rechromatographed on DEAE Sepharose CL-6B. Both activities co-eluted from the column but on dialysis were found in the diffusate. After desalting the diffusate with Sephadex G10, the presence of CMP-NANA was proved by mass spectrometry. This confirmed previous work and is the first unequivocal demonstration of CMP-NANA in constituents of human blood cells.

Blood Cells↗
PubMed1998

Exploring the mechanisms of antigen processing by cell fractionation.

It is becoming increasingly clear that most of the intracellular compartments that contain MHC class II products in antigen-presenting cells simply represent the conventional endosomes and lysosomes that are expressed in all cell types. Data from recent cell fractionation studies, however, predominantly those using electrophoresis techniques, show that a population of class-II-containing vesicles exists that may comprise a class of endosomes that are specialized for antigen processing. Strong support for this possibility comes from the observation that such specialized structures, designated class II vesicles (CIIV), are particularly abundant in mature dendritic cells.

Antigen Presentation↗
PubMed1985

Autoradiographical localization of luteinizing hormone releasing hormone (LHRH) receptors on rat testicular intertubular cells fractionated on Percoll density gradients.

The specific binding of 125I-labelled [D-Ser(tBu)6,des-GlyNH2(10)] LHRH ethylamide (LHRH-A) to testicular intertubular cells fractionated on Percoll density gradients was investigated. The greatest binding per cell occurred in the density region which contained the largest proportion of Leydig cells (sp. gr. 1.0820-1.0585). Autoradiographs of the cells from this region confirmed that silver stains were predominantly located over the Leydig cell, significantly (P less than 0.01) more grains were observed over this cell type in the total binding fractions than in the non-specific binding fractions. However, 5.9% of cells other than Leydig cells (testicular macrophages and indeterminate connective tissue cells) from this region also displayed significant displaceable binding (P less than 0.01). The location of [125I]LHRH-A binding to cells in other density regions, which did not contain identifiable Leydig cells, could not be established by autoradiography. These results confirm that the Leydig cell possesses LHRH receptors, but also indicate that other testicular cells have specific, high-affinity binding sites for LHRH-A, and may either be responsive to direct stimulation by LHRH, or may partially mediate the effects of LHRH and its agonists on Leydig cell function.

Animals↗
PubMed1963

Antigenicity of some Brucella melitensis cell fractions.

Glenchur, Harry (Minneapolis Veterans Administration Hospital, and University of Minnesota School of Medicine, Minneapolis), Ulysses S. Seal, Horace H. Zinneman, and Wendell H. Hall. Antigenicity of some Brucella melitensis cell fractions. J. Bacteriol. 85:363-368. 1963.-The components of a Brucella melitensis strain were obtained by differential centrifugation and diethylaminoethyl-cellulose ion-exchange chromatography. Rabbit antisera to these Brucella fractions were tested for agglutinins, precipitins, and blocking phenomena. Dermal hypersensitivity was also determined in rabbits. Insoluble (cell-wall) fractions provoked agglutinins, blocking antibodies and skin sensitivity, and precipitins for soluble antigens. Most soluble antigen fractions were capable of producing all the secondary immunological phenomena.

Antigens↗
PubMed1978

Cell fractionation and cytological analysis of human lymphatic cells from tonsil and blood.

No differences could be detected in the discontinuous density gradient when comparing the distribution pattern of tonsillar cells and peripheral lymphocytes. When the Kiel nomenclature is applied, the presence of centrocytes, centroblasts, lymphocytes and plasma cells in the tonsil is shown by a cytological analysis. The biological importance of these results is discussed from the point of cellular proliferation and tonsillar function.

Adult↗
PubMed1997

Lecithin:retinol acyltransferase and retinyl ester hydrolase activities are differentially regulated by retinoids and have distinct distributions between hepatocyte and nonparenchymal cell fractions of rat liver.

The cellular distribution of enzymes that esterify retinol and hydrolyze retinyl esters (RE) was studied in liver of vitamin A-sufficient, -deficient, and deficient rats treated with retinoic acid or N-(4-hydroxyphenyl)-retinamide. Livers were perfused and cell fractions enriched in hepatocytes, and nonparenchymal cells were obtained for assays of RE and enzyme activity. The specific activity of lecithin:retinol acyltransferase (LRAT) was approximately 10-fold greater in the nonparenchymal cell than the hepatocyte fraction from both vitamin A-sufficient and retinoid-treated rats. Total RE mass, newly synthesized [3H]RE and LRAT activity were positively correlated in liver and isolated cells of both normal (P < 0.0001) and retinoid-treated rats (P < 0.0002). In nonparenchymal cells, these three constituents were nearly equally enriched as evaluated by their relative specific activity values (RSA, defined as the percentage of recovered activity divided by the percentage of recovered protein), which were each significantly greater than 1.0, with values of 4.3 for total RE mass (P < 0.05), 3.6 for newly synthesized [3H]RE (P < 0.01) and 3.8 for LRAT activity (P < 0.01). In contrast, the specific activities of neutral and acid bile salt-independent retinyl ester hydrolases (REH) did not vary with vitamin A status, and their RSA values were close to 1.0 in both hepatocytes and nonparenchymal cells. These data show that LRAT and REH are differentially regulated by retinoids and that these enzymes also differ in their spacial distribution between liver parenchymal and nonparenchymal cells.

Acyltransferases↗
PubMed1979

Distribution of newly formed ribosomal proteins in HeLa cell fractions.

The distribution of newly formed ribosomal proteins between cytoplasmic, nucleoplasmic, and nucleolar fractions of HeLa cells was determined. All but a few of the newly formed ribosomal proteins were concentrated 10- to 50-fold in the nucleolus and two- to fivefold in the nucleoplasm. Nevertheless, substantial amounts were found in the cytoplasm. Pretreatment of cells with actinomycin D to deplete the nucleolar pool of ribosomal precursor RNA had no effect on the concentration of newly formed ribosomal proteins in the nucleus, but did lead to an increased amount in the nucleoplasm at the expense of the nucleolus.

Cell Nucleolus↗
PubMed1969

Preparation of cell walls and protoplasm of Neisseria with the Ribi cell fractionator.

The varied pressures required for disruption of Neisseria gonorrhoeae and other species of Neisseria when the Sorvall-Ribi refrigerated cell fractionator is used in the preparation of cell walls and cellular protoplasm are reported. Optimal disruption pressure for the gonococcus was considerably less than that required for other members of the genus Neisseria. Pressures varied from 8,000 psi for N. gonorrhoeae F62, colony type 4, to 22,000 psi for the nonpathogenic Neisseria-N. sicca, N. flava, and N. catarrhalis. Representative electron photomicrographs are shown.

Bacteriological Techniques↗
PubMed1986

Characterization of early compartments in fluid phase pinocytosis: a cell fractionation study.

Flotation through a 5.6% Percoll gradient of pinosomes from Chinese hamster ovary (CHO) cells labelled during a 10 min internalization period with horseradish peroxidase (HRP), a solute, revealed two pinosomal populations, the expected low-buoyancy population and an unexpected buoyant population. The buoyant pinosomes that sedimented similarly to plasma membrane were not an artifact of HRP trapping during homogenization or of cell surface-adherent HRP. No trapping or cell surface adherence of HRP could be detected by biochemical or cytochemical assays, even after internalization periods as short as 15 s to 1 min. With short uptake times, the buoyant pinosome population was the major HRP positive vesicle population, suggesting a precursor-to-product relationship between the two populations. In pulse-chase experiments, the buoyant pinosome population was shown to be highly exocytic and the precursor to later pinosomes. By electron-microscope cytochemistry, rapidly labelled, HRP positive pinosomes (15 s to 1 min uptake) were typically smooth vesicles with a median diameter of approximately equal to 0.30 micron and a size range from approximately equal to 0.10 micron to greater than 1.0 micron in diameter. We suggest that these rapidly labelled structures are a very early stage in the intracellular processing of pinocytic vesicles.

Animals↗