Core D will provide the techniques and morphologic expertise for the studies proposed in this SCOR Program. Services include the personnel, supplies and facilities for performing light and electron microscopy, immunohistochemistry on tissue sections and bronchoalveolar lavage cells, autoradiography and in situ hybridization. Quantitative techniques will be applied tumor necrosis factor-a (TNF-a), interleukin-8, endothelin-1 (ET-1) and PGH synthase will be examined in BAL cells from patients with the Adult Respiratory Distress Syndrome (ARDS). In situ hybridization with probes to each peptide will also be used to demonstrate alterations each of the genes. Such data will indicate whether TNF antibodies and glutathione ethyl esters alter regulation of these peptides. In addition, we will provide routine histological examination of all available autopsy lungs and lung biopsy tissue from these patients. This will allow correlations between the structural changes in our animal models and patients with ARDS. In we will examine the morphological alterations in the lung following administration of TNF-a, PGH synthase and ET-1 in the presence and absence of a TNF-a antibody, a neutrophil adhesion blocker, NPC15669, and the glutathione ethyl ester, GSE. These data will allow correlation between structural, functional and biochemical changes and provide insights into the efficacy of each """"""""protective"""""""" agent. We will use in situ hybridization to localize xanthine oxidase mRNA in ling and liver of sheep during endotoxemia. In project V. we will examine lung, liver and kidney tissues for evidence of injury following administration of various forms of liposomes, examination of DNA/liposome complexes by scanning microscopy, localization of liposomes in organs using a fluorescent marker (5,6- carboxyfluorescein), localization of cDNA's in tissue by in situ hybridization, visualization of reporter genes such as chloramphenicol acetyltransferase (CAT) and beta-galactosamine, as well as localization of PGH synthase by immunohistochemical techniques. The techniques and interpretative skills of this Core are crucial to the successful completion of this SCOR application.
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