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Biomedical subjects

J Gouranton

Publications and source records attributed to J Gouranton.

At least 19 recordsLinked to original sources

Expression of RNA isolated from the water-shunting complex of a sap-sucking insect increases the membrane permeability for water in Xenopus oocytes.

The highly specialized membranes of the filter chamber found in the digestive tract of some homopteran insects could represent a favorable material for characterizing water channels. In order to demonstrate that membrane proteins of this epithelial complex serve as water channels, we have investigated the membrane permeability for water in Xenopus oocytes injected with RNA isolated from the filter chamber. Volumes of oocytes injected with filter chamber RNA were increased by 15% following a 16-min osmotic shock, while volumes of oocytes injected with RNA from midgut not of filter chamber or with water were increased only by 8.5 and 10%, respectively. This significant difference in oocyte swelling leads us to conclude that RNA isolated from the filter chamber contains mRNA coding for water channel proteins.

Animals

Electron microscopic observation of the respiratory tract of SPF piglets inoculated with Mycoplasma hyopneumoniae.

Seven hysterectomy derived piglets were repeatedly challenged with Mycoplasma hyopneumoniae during the first week of life. Samples of trachea, bronchi and lung tissue collected 2-11 weeks post-inoculation (p.i.) were examined using light and electron microscopy. Autoradiography was used to study in more detail the site of M. hyopneumoniae multiplication. Gross lesions were observed in lung tissue and were characterized by hyperplasia of the epithelium and an increased mononuclear cell accumulation in perivascular and peribronchiolar areas. Mild lesions of the trachea and the bronchi, including epithelial hyperplasia and infiltration of the lamina propria by inflammatory cells, were noted. Electron microscopy showed that, 2-6 weeks p.i., changes in the mid-trachea and bronchi surface consisted of the loss of cilia. Mycoplasmas covered tufts of cilia remaining on the epithelial cell surface. Scanning and transmission electron micrographs showed that they were predominantly found closely associated with the top of cilia. No specialized terminal structure could be seen and no mycoplasma cells were identified lying free in the lumen nor in close contact with the plasma membrane of cells or microvilli. Some fine fibrils radiating from one mycoplasma to another or to cilia were seen at higher magnification by scanning electron microscopy. Six to eleven weeks p.i., a disrupted epithelial surface lacking cilia was observed. Cells were desquamated and shed into the lumen with cellular remains containing droplets of mucus. Autoradiography revealed that label corresponded to the observed mycoplasma distribution. At the top of cilia, a high density of labeling was visible in the zone of high mycoplasma concentration. Therefore, incorporation of the label in the mycoplasma is proof or their multiplication in the trachea. The intimate association between the mycoplasma and cilia may be an important factor in the pathogenesis of the disease caused by M. hyopneumoniae (swine enzootic pneumonia).

Animals

Structural and biochemical observations on specialized membranes of the "filter chamber", a water-shunting complex in sap-sucking homopteran insects.

Many homopteran insects feed on plant sap which contains solutes in very low concentration. Their digestive tract presents a complex called the "filter chamber" where the excess dietary water is believed to flow directly from the initial part of the midgut to the terminal part of the midgut and the proximal regions of the Malpighian tubules. Freeze-fracture experiments carried out on the filter chamber of Cicadella viridis revealed the presence of intramembrane particles on the whole surface of the microvilli and of basal membrane infoldings of the cells. Examination of negatively stained isolated membranes and of freeze-dried shadowed membranes revealed that the inner surface of the membrane is covered with particles protruding into the cytoplasm; they correspond to the numerous intramembrane particles observed on the P fracture face of the membrane. The outer surface of the membrane exhibits a regular network which corresponds to that observed on the E fracture face. SDS-PAGE analyses were performed on purified membranes of the filter chambers of C. viridis and Philaenus spumarius. In both cases 2 major components, 25 kDa and 75 kDa, were detected. These 2 components appear to be specific for the filter chambers since they were not found in membranes isolated from the other parts of the midgut. Thus, the membranes of these filter chambers, thought to be water-shunting complexes, possess structural and biochemical peculiarities which are probably related to water permeability.

Animals

Characterization of a ferritin isolated from the midgut epithelial cells of a homopteran insect, Philaenus spumarius L.

Crystalline accumulations of ferritin-like particles are present within the cytoplasma and the nucleus in midgut epithelial cells of the homopteran Philaenus spumarius. A structural study at the electron microscope level reveals that these particles have the morphological characteristics of the ferritin molecule: crystals have a face-centered cubic structure with a lattice parameter of 14 +/- 1 nm; negatively stained isolated particles have the appearance of ferritin; on rotary-shadowed particles 3 axes of symmetry are clearly seen; image processing performed on selected molecules demonstrates a 4-fold symmetry. A semiquantitative electron microprobe analysis effected on aggregates of microcrystals in thin sections reveals a high atomic ratio Fe/P. Analyzed by SDS-PAGE, the protein subunit has a molecular weight of 18,600. The amino acid composition of the protein bears the general characteristics of the ferritin molecule in terms of polar and nonpolar residues. But in terms of sequences, this protein displays a strong dissimilarity to rat liver ferritin as demonstrated with a common amino acid index test and with immunoelectrophoresis experiments.

Animals

Flow cytometric quantitative evaluation of phagocytosis by human mononuclear and polymorphonuclear cells using fluorescent nanoparticles.

The use of fluorescent polymethacrylic nanoparticles (0.3 micron) as a flow cytometric reagent in the quantitative evaluation of phagocytosis by human mononuclear and polymorphonuclear cells is described. The preparation of the nanoparticles, by emulsion copolymerization of methacrylic monomers, and their physicochemical properties are briefly summarized. Nanoparticles coupled with a fluorescent agent (ethidium bromide) were used in a flow cytometric assay to study opsonin-independent phagocytosis by human polymorphonuclear cells and by human monocytes. The phagocytosis of nanospheres by monocytes was determined by flow cytometry from the fluorescence distribution and ingestion was visualized by scanning and transmission electron microscopy. One possible application of the fluorescent nanoparticles is the simultaneous analysis of cell surface antigens and cell phagocytic activity.

Acrylates

[Relation between cyclic AMP and phagocytosis in human monocytes].

The involvement of cyclic adenosine 3'-5' monophosphate (cAMP) in the regulation of human monocyte phagocytosis of staphylococcus aureus in vitro was demonstrated by assay of adenylate cyclase (AC) and cAMP phosphodiesterase (PDE). Phagocytosis was associated with diminished AC activity (p less than 0,05) and concurrently increased PDE activity (p less than 0,005). Transmission electron microscopy provided evidence that these changes coincide with peak phagocytic activity occuring 10-20 minutes after the start of phagocytosis.

3',5'-Cyclic-AMP Phosphodiesterases

Changes in cAMP metabolism during phagocytosis of S. aureus by human monocytes.

Involvement of cyclic adenosine 3',5'-mono-phosphate (cAMP) in the phagocytosis of staphylococci by human monocytes was demonstrated by assay of adenylate cyclase and cAMP phosphodiesterase (PDE). Monocyte adenylate cyclase and PDE activities were assayed on cell homogenates prepared from monocytes isolated by plate adherence. Phagocytosis of staphylococci was associated with diminished adenylate cyclase activity, which reached a minimum after 10 min of incubation (p less than 0.05), and an increase in PDE activity (p less than 0.005). Transmission electron microscopy provided evidence that these changes coincide with peak phagocytic activity occurring between 10 and 20 min after the start of phagocytosis but could not be assigned to a particular stage in the phagocytic process.

3',5'-Cyclic-AMP Phosphodiesterases

The intranuclear filamentous inclusions of a human glioma. Their relation with nuclear bodies.

The intranuclear filamentous inclusions of a human glioma were analysed with an electron microscope equipped with a goniometer stage. The inclusions consist of 6 to 8 filaments. Considering the organization of the constituent filaments we distinguish three basic types: 1. Filamentous bundles of more or less parallel filaments, forming a cigarshaped inclusion. 2. Crystalloid inclusions: a. Prisms. They consist of stacked layers of strictly parallel filaments. The angle formed by the filaments of adjacent layers if 60 degrees. b. Cylinders. The layers of filaments are bent up and may form either a circle or a spiral, when the inclusion is seen in cross-section. 3. Partially crystalloid or "intermediate" inclusions. We consider them to be transitional forms between types 1 and 2 inclusions. The crystalloid layers of such intermediate inclusions may form either prisms or cylinders. Finally, the similarity between the granulo-fibrillar capsules surrounding granular nuclear bodies and filamentous inclusions, as well as the existence of granular material dispersed between the filaments of some inclusions led us to investigate a relationship between these two structures.

Brain Neoplasms

Observation on intranuclear crystal and nucleolar size at different stages of cell differentiation in the midgut epithelium of several insects.

Based on an inverse size relationship between nuclear crystal and nucleolus in different cells it has been postulated by several authors that the crystal develops from nucleolar materials. The purpose of the present paper is to investigate the validity of this argument. Intranuclear proteinaceous crystals appear in differentiating midgut cells of Gyrinus marinus and Tenebrio molitor. In an autoradiographic study we have previously demonstrated in these two species that the crystals do not develop from nucleolar materials. However, an inverse relationship with regard to size is observed between these 2 structures during the cell differentiation: the cross-sectional area of the nucleolus decreases when the cross-sectional area of the crystal increases. But a decrease in size of the nucleolus is also observed during the differentiation of the midgut cells of Gyrinus natator where the crystals are not present. Consequently an inverse size relationship cannot be a sufficient argument to postulate that intranuclear crystals and nucleoli are interconvertible structures; decrease in size of the nucleolus is not related to development of the intranuclear crystal.

Animals

[Composition, structure and mode of formation of mineral concretions in the midgut of the Cercopid (Homoptera)].

The granules which occur in the cells of a part of the midgut wall in Cercopid larvae and adults (Homoptera) have been studied by biochemical and cytochemical methods and by electron microscopy. The granules have a diameter up to about 2micro and contain calcium, magnesium, iron, carbonates, and phosphates. Protein and acid mucopolysaccharide have also been detected. A chromatographic study shows that uric acid and guanine are not present. The young concretions occur primarily in ergastoplasmic cisternae. They are first wholly electron-opaque, but their center becomes more and more clear. In very old spheres, only a thin shell of electron-opaque material remains. The spheres which have reached about 1micro in diameter are all associated with myelin figures. The granule-containing cells, which nearly occlude the lumen of the midgut in larvae, are eliminated in the very young adults, but the storage excretion still continue in adults.

Animals