Biomedical subjects
Publications and source records attributed to F Gil.
Ajoene, a garlic-derived compound that prevents platelet activation, inhibited the growth of Paracoccidioides brasiliensis, a fungal pathogen for humans, by affecting the integrity of the fungal cytoplasmic membrane. This action may be the basis for the study of ajoene as a possible specific antifungal drug.
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Glutathione metabolism was studied in isolated hepatocytes from 48-h starved rats. Phenylephrine (10 microM, final concentration) was incubated in the presence of a mixture of L-glutamine, glycine, L-serine, and L-methionine (at 10 times their normal plasma concentration). Alpha-adrenergic stimulation provoked a decrease in glutathione (GSH) synthesis. This effect was accompanied by an enhanced efflux of glutathione from the cells. Phenylephrine stimulated the rate of glutathione disulfide (GSSG) formation; however, this effect was clearly insufficient to explain the disappearance of GSH. Our results suggest that the decrease in cellular GSH levels observed under conditions of shock, stress, or peripheral inflammation can be explained by a dual effect, i.e., an increase in glutathione efflux and an inhibition of its synthesis.
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Isoflurane inhibits oxidative metabolism of halothane. Because hepatotoxicity of chemicals may be associated with their metabolism, whether isoflurane can protect the liver against chemical injury was investigated. Hepatic injury was produced in female F344 rats by a 30-minute exposure to 250 ppm of carbon tetrachloride. In this and all other parts of the study, the inspired oxygen concentration was maintained at 21%. The injury was accompanied by elevated activity of liver enzymes in serum (SGOT, SGPT, and SDH), enlarged liver, fatty infiltration of the liver, and vacuolar degeneration of hepatocytes. These signs of toxicity were partly or completely suppressed by concurrent exposure to subanesthetic concentrations of isoflurane (0.2 or 0.038%, respectively). The protective effect was concentration-dependent. Enflurane was protective, but less so than isoflurane. Nitrous oxide and fentanyl had no protective effect.
Early steps of replication (penetration and uncoating) of the OSU strain of porcine rotavirus were studied in MA-104 cells. After adsorption of trypsin-treated viruses at 4 degrees, followed by a shifting of the temperature to 37 degrees, particles were seen within coated pits, coated vesicles, and secondary lysosomes, indicating that virus entry occurred by receptor-mediated endocytosis, and that uncoating (removal of the outer capsid) could occur by the effect of lysosomal enzymes. This latter aspect was studied using lysosomotropic drugs (chloroquine and ammonium chloride), which were found not to inhibit rotavirus replication, indicating that the low intralysosomal pH is not responsible for virus uncoating. The effect of Ca2+ concentration on intracellular rotavirus uncoating was investigated by treating cells with the calcium ionophore A23187, to increase the intracellular Ca2+ concentration during the early stages of virus replication. Under these conditions rotavirus uncoating did not occur, suggesting that the low Ca2+ concentration in the intracellular microenvironment may be responsible for rotavirus uncoating.
Glutathione (GSH) decreases in dog myocardium upon acute coronary occlusion when compared with sham-operated dogs. Total glutathione content (GSHeq = GSH + 2GSSG) remains unchanged throughout the experiment (6 h after surgery) in both sham- and acute coronary occlusion-operated dogs. GSSG and GSH/GSSG ratio increases and decreases respectively in all animals but tends to reach the normal value after 6 h in sham-operated dogs. Both parameters (GSSG and GSH/GSSG ratio) remain altered in acute coronary occlusion-operated ones. This alteration of glutathione status in ischemic myocardium is discussed.
Human epidermoid carcinoma A431 cells attach more rapidly to collagen type-I and -IV substrates than to surfaces coated with laminin or fibronectin. The diminished intercellular interaction and rounding up manifested when these cells are exposed to epidermal growth factor (EGF) in tissue culture plastic or collagen films is not shown when the assay is performed on 3-dimensional collagen. In the latter substrate, cells exposed to EGF reveal greater cell cohesion with interdigitations and desmosomal junctions, compared to the limited intercellular interaction detected in similarly treated cells assayed in tissue culture substrate. Although the protein synthesis inhibitor, cycloheximide, did not prevent these EGF-mediated changes, metabolic labelling indicated a substrate-dependent effect of EGF on the synthesis of proteins associated with the Triton-insoluble cytoskeletal matrix. The involvement of cytoskeleton components in the EGF effects on intercellular adhesion was shown by its susceptibility to cytochalasin B, known to disorganize actin-containing microfilaments. Some of the mechanisms of epithelial morphogenesis and the influence of extracellular matrix components on cell-receptor/growth-factor interactions may now be suitably analyzed by examining the EGF effects on A431 cells grown on different substrata.
The inner capsid structure of the OSU strain of porcine rotavirus was studied by electron microscopy of freeze-dried preparations and of negatively stained chemically disrupted virus particles. The analysis of the particles by the freeze-drying technique revealed a T:13 l (laevo) symmetry for the organization of the inner capsid. Treatment of single-capsid rotavirus particles with 30% formamide or 5 M-urea resulted in their degradation, giving rise to very similar products, corresponding to isolated vertices, edges and faces of the virus icosahedron. An analysis of such structures confirmed the triangulation number and handedness of the rotavirus inner capsid, and provided evidence for the open-mesh model, in which the five- and six-coordinated axes are represented by 'holes' formed by smaller trimeric morphological subunits.
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B16 melanoma cells exhibited decreased adhesion to substration and the presence of cytoplasmic granules, resembling lipid inclusions, when cultured in vitro in the presence of syngenic serum from C57/BL6 mice. This constrasted with the rapid cell attachment and absence of cytoplasmic granules in cultures seeded in medium supplemented with an identical concentration of fetal bovine serum. Electrophoretic comparison of intracellular proteins revealed similar patterns in detergent-soluble and matrix-associated proteins from cells grown with bovine or mouse serum. However, a similar analysis of the conditioned media showed clear differences in methionine-rich species which migrated in the 100 kd region in cells grown with bovine serum, and as 110 kd component in cells grown with mouse serum. Our data indicate that the poor attachment to substratum and changes in cytoplasmic organization of B16 melanoma cells, is primarily associated with specific changes in secreted proteins.
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Thirty-four strains corresponding to 16 species of Clostridium were examined for growth inhibition activity and production of phage and bacteriocinlike particles. Twenty-five strains were found to inhibit growth of one or more of the other strains. The widest range of activity corresponded to C. bifermentans 18137 which inhibited growth of 23 strains. Cell-free filtrates of positive cultures were inactive with the exception of the one from C. bifermentans 18137. A wide variety of particles with diverse morphology was observed in lysates of cultures treated with mitomycin C (1 microgram/mL).
Human rotaviruses were capable of efficient multiplication in LLC-MK2 cells when the inoculum was pre-treated with trypsin, centrifuged on to the cell monolayer and the infected cells maintained in a medium containing trypsin. However, not all of the human rotavirus isolates used to infect cells resulted in efficient virus production. The ability of human isolates to multiply in cultured cells was studied by direct observation of virus in the electron microscope, by radioactive labelling with 3H-uridine of the newly synthesized virus and by electron microscopic examination of thin sectioned infected cells. With one of the specimens used (F-617) only 5 to 10% of the cells showed evidence of virus multiplication, with the great majority of the infected cells showing numerous complete (double-capsid) virus particles scattered in the cytoplasm. When cells were inoculated with another human specimen (SIB-I), infected cells were more abundant, reaching a maximum of 60%; however, a variety of particle types, probably representing different subviral structures or different steps of rotavirus morphogenesis, were commonly observed. The presence of these aberrant or incomplete virus structures may represent a manifestation of the defectiveness of this virus and may explain the difficulties encountered in its serial passage.
Selected functions of polymorphonuclear leukocytes were studied in patients with paracoccidioidomycosis (South American blastomycosis), in healthy control individuals, and in patients with diseases unrelated to paracoccidioidomycosis. Patients with paracoccidioidomycosis were also evaluated by standard immunological techniques. Phagocytosis and digestion of Paracoccidioides brasiliensis yeastlike cells in vitro was estimated by an original method. It was based on the appearance of phagocytosed P. brasiliensis in preparations stained by a modification of the Papanicolaou method and examined with phase-contrast optics. Interpretation of such findings was confirmed by electron microscopy. Two strains of P. brasiliensis were used. Strain 8506 was freshly isolated from a patient. Strain Pb9 was known to be nonpathogenic and to have a peculiar cell wall composition. Yeastlike cells of the Pb9 strain were digested significantly better than those of strain 8506. A higher number of leukocytes per fungus cells led to a higher proportion of digested P. brasiliensis. Leukocytes from patients with paracoccidioidomycosis phagocytosed the fungus in a normal way, but had a significant lower ability to digest it in vitro. When individual cases were analyzed, there was an excellent correlation between clinical evolution and digestive ability of polymorphonuclear leukocytes. There was good correlation between both of these and immunological parameters. Leukocytes from all groups behaved comparably in tests of general leukocyte function and in their abilities to kill and digest Candida albicans. Our results indicate that, as a group, polymorphonuclear leukocytes from patients with paracoccidioidomycosis had a significant, rather specific, defect in their in vitro digestive capacity against phagocytosed P. brasiliensis. There was also an inverse correlation between strain pathogenicity and its susceptibility to in vitro digestion by polymorphonuclear leukocytes. Our findings are concordant with and relevant to clinical reality.
Electron microscopic examination of an Aedes pseudoscutellaris mosquito cell line (Mos. 61) revealed the presence of a large number of virus-like particles (VLP) in the cytoplasm of approximately 10% of the cells. These particles have a diameter of 36 nm, do not contain a lipid envelope, and have a buoyant density of 1.40 g/ml in CsCl. VLP contain DNA which appears to be single-stranded by SU nuclease assay. PAGE of the VLP demonstrates the presence of four structural polypeptides with molecular weights of 52K, 70K, 75K and 100K daltons, the smaller one being the majro polypeptide species and accounting for 71% of the the total protein mass. The A. pseudoscutellaris VLP appear to share some, but not all, of the characteristics of the genus Densovirus, family Parvoviridae. Their possible biologic importance and taxonomic location are still unclear.
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