The collagen architecture in normal, aging, and osteoarthritic articular cartilage was studied optically using a new silver staining technique based on specimens from 50 autopsy cases, four amputated limbs, and six osteoarthritic knees. In the normal articular cartilage, the collagen fibrils in the superficial zone were compactly arranged into layers of decussating flat ribbons mostly parallel to the artificial split lines. The fibrils showed a tendency to condense into vertical arcade columns undergirded by tangential bundles in the intermediate zone. In the deep zone, the fibrils formed a random meshwork with a slight preponderance of vertical fibrils in the perilacunar region. Three types of early degradative lesions involving the collagen network were identified. Type I lesions consisted of focal superficial disruptions related to age and friction. Type II lesions consisted of focal disruptions of tangential fibrils in the intermediate zone leading to cyst formation, probably representing a form of local stress failure. Type III lesions were found in the patella and consisted of marked swelling of the superficial zone, the cause of which was unknown. Lesions of varying severity were seen within each of the three types; the morphological changes of the more severe lesions overlapped with those of clinically overt osteoarthritis.
This study was aimed to establish embryonic stem (ES)-like cells from blastocysts derived from somatic cell nuclear transfer (SCNT) in pig. Somatic cells isolated from both day-30 fetus and neonatal cloned piglet were used for donor cells. A total of 60 blastocysts (46 and 14 derived from fetal and neonatal fibroblast donor cells, respectively) were seeded onto a mitotically inactive mouse embryonic fibroblast (MEF) monolayer and two ES-like cell lines, one from each donor cell type, were established. They remained undifferentiated over more than 52 (fetal fibroblast-derived) and 48 (neonatal fibroblast-derived) passages, while retaining alkaline phosphatase activity and reactivity with ES specific markers Oct-4, stage-specific embryonic antigen-1 (SSEA-1), SSEA-4, TRA-1-60 and TRA-1-81. These ES-like cells maintained normal diploid karyotype throughout subculture and successfully differentiated into embryoid bodies that expressed three germ layer-specific genes (ectoderm: beta-III tubulin; endoderm: amylase; and mesoderm: enolase) after culture in leukemia inhibitory factor-free medium. Microsatellite analysis confirmed that they were genetically identical to its donor cells. Combined with gene targeting, our results may contribute to developing an efficient method for producing transgenic pigs for various purposes.
A case of malignant carcinoid tumor (neuroendocrine carcinoma) of the cecum is described. The neoplasm arose in a patient who had chronic ulcerative colitis for 16 years. There is no previous documentation of this combination, which is surprising, in view of the known association of colonic adenocarcinomas and lymphomas with ulcerative colitis. The reasons for this discrepancy are discussed and it is postulated that ultrastructural examination of poorly differentiated colonic neoplasms may result in a more frequent identification of this association.
Mucosal biopsies from four women with collagenous colitis and ten controls were studied. By light microscopy the cells of the pericryptal fibroblast sheath appeared diminished in number but increased in size in collagenous colitis. Electron optically in the controls the pericryptal fibroblasts were in intimate contact with the epithelial basal lamina in the crypts. On the free surface the fibroblasts maintained contact with the epithelial cells by attenuated cell processes. In collagenous colitis, in the middle and upper thirds of the crypts the fibroblasts sheath was separated from the epithelium and the fibroblasts assumed the characteristics of myofibroblasts. The separation was accentuated towards the mouths of the crypts. Beneath the surface epithelium the attenuated fibroblast cell processes seen in normal colon were grossly deficient. The basal lamina was also deficient focally and the surface epithelial cells were resting directly on a thickened collagen table. In collagenous colitis the excess collagen appeared to be secreted by the activated myofibroblasts of the pericryptal sheath.
The present study was undertaken to study the mechanism of cyclosporine-induced nephrotoxicity and hypertension. Cultured rat microvascular endothelial cells were exposed to cyclosporine for up to six days at one of the following concentrations: 10, 50, 250, and 1000 ng/ml of culture medium. Cyclosporine inhibited endothelial cell replication in a dose-dependent manner; at higher concentrations (250 and 1000 ng/ml), cell replication decreased by as much as 70 to 90% of controls at four and six days post-treatment, with no evidence of increased cell death. Drug-treated endothelial cells revealed abnormal morphological changes such as formation of cytoplasmic vesicles and nucleolar changes. Prostacyclin release by endothelial cells was increased by about threefold with the addition of cyclosporine (P less than 0.01). Indomethacin significantly decreased prostacyclin release by endothelial cells in the presence or absence of cyclosporine (P less than 0.01). Our data suggest that cyclosporine-induced nephrotoxicity may be mediated, at least partly, by the inhibitory influence of cyclosporine on the regenerative response of microvascular endothelial cells to injury, and the resultant alterations in prostacyclin production by these cells.
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