Mannoproteins, as the main constituents of the outer layer of yeast cell walls, are able to interact with phagocytic cells in an opsonin-independent manner through the mannose receptor (MR) and to induce yeast ingestion by the professional phagocytes. Moreover, the MR also mediates endocytosis of soluble ligands through clathrin-coated pits. Here, we studied some aspects of the interaction between the MR and Candida albicans using murine E-clone macrophages and the consequences on MR trafficking. Using a pull-down assay involving mixture E-clone macrophage detergent lysate with mannosylated Sepharose beads and glutaraldehyde-fixed, heat-killed (HK) C. albicans, we found that binding of solubilized MR to mannosylated particles occurred with characteristics similar to the receptor's cell-surface mannose-binding activity. We then demonstrated that MR expressed on E-clone macrophages contributed to phagocytosis of unopsonized, HK C. albicans and that yeast phagocytosis induced a decrease in MR endocytic activity without concomitant degradation of the receptor in the time lapse studied.
The early events in the interaction of two polyene (amphotericin B and nystatin) and five imidazole (clotrimazole, ketoconazole, miconazole, isoconazole, and econazole) antimycotics used at fungicidal concentrations with the surface of Candida albicans were studied by scanning electron microscopic examination of treated intact young yeast cells, treated spheroplasts, and spheroplasts liberated from treated young yeast cells. In all cases, treatment lasted 2 h. The polyenes passed through the yeast cell wall and interacted with the cytoplasmic membrane causing the spheroplasts to lose their characteristic spheric form and to liberate their contents. Clotrimazole caused the formation of numerous circular openings in the cytoplasmic membrane, but only when the agent was used to treat spheroplasts directly. Ketoconazole, miconazole, isoconazole, and econazole interacted with the cell wall causing formation of convolutions and wrinkles. The three imidazole derivatives that are structurally closely related, miconazole, isoconazole, and econazole, inhibited the enzyme-catalyzed release of spheroplasts from young yeast cells.
The ability of Succhuromyces boulardii to protect mice against intestinal pathology caused by toxinogenic Clostridiurn dzBcile was studied. Different regions of the intestine of experimental mice were prepared for observation by scanning electron microscopy or homogenized for C. dzBcile enumeration and quantification of toxin A by enzyme immunoassay and toxin B by cytotoxicity. The test group was treated for 6 d with an S.boulurdii suspension in drinking water and challenged with C. dz&le on day 4. The three control groups were: axenic mice, mice treated with only S. boulurdii and mice only challenged with C. dz@cile. The results showed that: (i) 70% of the mice infected by C. dzjicile survived when treated with S. boulurdii; (ii) the C. dzBcile-induced lesions on the small and large intestinal mucosa were absent or markedly less severe in S. boulurdii-treated mice; and (iii) there was no decrease in the number of C. dzBcile but rather a reduction in the amount of toxins A and B in S. boulardii-treated mice.
Caspofungin is a member of the echinocandin class of antifungal compounds that inhibit 1,3-β-d-Glucan synthase. As patient exposure to caspofungin (CAS) broadens, the number of infecting strains with reduced susceptibility to this drug is expected to rise. In the present study, the in vitro effects of varying concentrations of CAS against Candida albicans isolates presenting reduced susceptibility to CAS were studied in comparison with a reference strain. Two C. albicans isolates presenting high minimal inhibitory concentrations (MIC = 8 μg ml(-1) ) were selected: one isolate obtained in the laboratory under continuous antifungal selection pressure (CaIn-R) and one clinical isolate (CaClin-R) from a patient with a therapeutic failure. Results showed that after 24 h of CAS exposure, CaIn-R and CaClin-R presented a partial growth inhibition in comparison with the reference strain. Moreover, scanning electron microscopy and transmission electron microscopy studies showed that the cell walls of CaIn-R and CaClin-R were less altered than that of the reference strain. These observations suggested that although CaIn-R and CaClin-R cells were misshapen after CAS exposure, cell lysis was limited after 24 h of treatment indicating higher survival ability for CaIn-R and CaClin-R in the presence of CAS.
We conclude from this long-term experience that during ECPII, catheter blockage remains the major recurring complication, probably involving a local immune-inflammatory response in the peritoneum.
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