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  • 1
    ISSN: 1573-904X
    Keywords: tracheal epithelial cell monolayer ; transport ; metabolism ; Gly-L-Phe ; rabbit ; aminopeptidase M
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-0603
    Keywords: transport ; epithelium ; edema ; beta-agonist ; barrier ; permeability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To investigate the cell physiologic and biological properties of the alveolar epithelium, we studied rat alveolar epithelial cell monolayers grown on permeable supports in primary culture. Type II alveolar epithelial cells were disaggregated using elastase, and partially purified on a discontinuous metrizamide gradient. These isolated cells were plated onto tissue culture-treated Nuclepore membrane filters at 1.5×106 cells/cm2 and maintained in a humidified incubator (5% CO2 in air, 37° C). After 2 days in culture, the bathing media on both sides of the cell monolayers were changed to fresh culture medium, thus removing nonadherent cells (mostly leukocytes). These monolayers exhibit a high transmonolayer resistance (〉2000 Ω-cm2) and actively transport ions. Radionuclide flux studies indicate that Na+ is the predominant ionic species absorbed actively under baseline conditions, accounting for about 80% of the total active ion transport. Cl− seems to be passively transported across the epithelium. However, when the epithelium is exposed to a beta-agonist (terbutaline), active absorption of Na+ is increased and active absorption of Cl− occurs. Although it is clear that both active Na+ and Cl− transport are dependent on Na+/K+-ATPase activity, and that Na+ enters cells predominantly through channels, the specific mechanisms by which Cl− enters and exits the alveolar epithelial cells remain unclear. The stimulated reabsorption of Na+ and Cl− may be important in helping to remove excess fluid from alveolar air spaces in the lung.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-904X
    Keywords: tracheal epithelial cell layer ; transport ; peptide transporter ; carnosine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. To determine the functional presence of a H+/peptide cotransport process in rabbit tracheal epithelial cell layers cultured at an air-interface and its contribution to transepithelial dipeptide transport. Methods. Rabbit tracheocytes were isolated, plated on Transwells, and cultured at an air-interface. After 5 or 6 days in culture, uptake and transepithelial transport of carnosine were examined. Results. Carnosine uptake by tracheocytes was pH-dependent and was saturable with a Michaelis-Menten constant of 170 μM. Moreover, carnosine uptake was inhibited 94% by Gly-L-Phe, 28% by (β-Ala-Gly, but not at all by Gly-D-Phe or by the amino acids β-Ala and L-His. Unexpectedly, transepithelial carnosine transport at pH 7.4 (i.e., in the absence of a transepithelial pH gradient) was similar in both the apical-to-basolateral (ab) and basolateral-to-apical (ba) directions. Lowering the apical fluid pH to 6.5 reduced abtransport 1.6 times without affecting ba transport, consistent with predominantly paracellular diffusion of carnosine under an electrochemical potential gradient. Conclusions. The kinetic behavior of carnosine uptake into cultured tracheal epithelial cell layers is characteristic of a H+-coupled dipeptide transport process known to exist in the small intestine and the kidney. Such a process does not appear to be rate-limiting in the transport of carnosine across the tracheal epithelial barrier.
    Type of Medium: Electronic Resource
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