Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.4.24.3 (collagenase)
18,340 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Fibronectin was localized in human cholesteatoma tissues by immunohistochemical methods. The avidin-biotin-peroxidase complex staining method was used with specific fibronectin antibody. Fibronectin appeared to be localized in the matrix of the cholesteatoma studied, particularly on the surface of the cell membranes and the nuclei of the basal cells and in connective tissue. Fibronectin was not seen in the granular layer or in the keratin area. Fibronectin was found on the surface of granulation tissue, mononuclear cells, fibroblasts and endothelial cells of blood vessels. These findings were confirmed by the immunofluorescent staining method. Our previous study showed that fibronectin induced a migration of keratinocytes, macrophages and fibroblasts demonstrated by the Boyden's chamber chemotaxis assay. Macrophages and fibroblasts were shown to produce collagenase, a bone resorption factor, in cholesteatomatous tissue. The present study showed the presence of fibronectin in the matrix of cholesteatoma and granulation tissue, suggesting that fibronectin might play an important role in the clinical development and invasive behavior of cholesteatoma.
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PMID:Localization of fibronectin in human middle ear cholesteatoma. 305 88

There is growing evidence suggesting that hepatic fat-storing cells (FSC) or Ito cells have an important function in vitamin A storage and metabolism and in the synthesis of connective tissue components in normal liver and during fibrogenesis. The purified FSC acquire a fibroblastic morphology and their vitamin A content decreases in culture. We cultivated cells under in vitro conditions that allowed the expression of FSC morphological and functional characteristics for 3-4 weeks of primary culture. Cells were isolated from rat liver by the collagenase-perfusion method without further purification and cultured with 3T3-conditioned medium, which seemed to stimulate the selective proliferation of the FSC. After 8-10 days, round and stellate cells grew actively from a few precursor cells in the primary culture and were not subcultivated; the stellate cells had the ability to become round and vice versa and were highly motile. The cells had intracytoplasmic lipid droplets, a well developed rough endoplasmic reticulum, Golgi complex, numerous vesicles filled with electron-dense material, and extracellular matrix (ECM) components on their surface. Both stellate and round cells showed the presence of desmin by immunofluorescence and vitamin A autofluorescence, but lacked peroxidase activity. The culture conditions we describe allowed the selective proliferation of cells with morphological and functional characteristics of the FSC in the normal liver, raising the possibility of studying FSC proliferation and differentiation.
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PMID:Culture of proliferating and differentiating fat-storing cells in 3T3-conditioned medium. 322 17

Kupffer cells and other sinusoidal cells were isolated after perfusion and incubation with pronase and collagenase of pieces of liver tissue obtained from organ donors. The resulting cell preparations contained endothelial cells, Kupffer cells and fat-storing cells as well as considerable numbers of leucocytes. Attempts to purify the different sinusoidal cell types by density centrifugation and centrifugal elutriation were successful only for Kupffer cells. Kupffer cells, in contrast to endothelial cells and fat-storing cells, could be kept in maintenance culture for at least 5 days. Cultured Kupffer cells were active in the endocytosis of foreign substances, such as colloidal carbon, latex beads, horseradish peroxidase and bacterial endotoxin. The cultured Kupffer cells synthesized and secreted considerable amounts of prostaglandins PGE2, PGF2 alpha, 6-keto-PGF1 alpha and thromboxane B2. The production of prostaglandins was influenced by the presence of Escherichia coli endotoxin.
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PMID:Isolation and culture of Kupffer cells from human liver. Ultrastructure, endocytosis and prostaglandin synthesis. 327 5

To investigate which cells of the liver express the receptor for transferrin, isolated rat liver cells produced by collagenase perfusion were fractionated by repeated differential centrifugation to produce hepatocytes (95% + 1%, mean +/- SD, n = 4) and nonparenchymal cells (97% + 1%, n = 3). Saturable, high-affinity binding of 125I-transferrin was demonstrated on intact cells at 4 degrees C, with average receptor numbers 20,900 +/- 3,160 (mean + SD, n = 4) for hepatocytes and 5,500 + 1,520 (n = 3) for nonparenchymal cells. Total cellular receptors measured in detergent permeabilized hepatocytes were 42,000 +/- 18,330 (mean +/- SD, n = 3) per cell and 14,760 +/- 7,120 (n = 3) per cell in the nonparenchymal fraction. Immunocytochemical demonstration of transferrin using antitransferrin, peroxidase antiperoxidase complex confirmed that both cell types bound transferrin. There was heterogeneity of the staining reaction since there was no detectable staining on 40% of hepatocytes and 60% of nonparenchymal cells. Microdensitometric analysis of the staining product corroborated the biochemical evidence that hepatocytes have, on average, more than three times more transferrin receptors than do nonparenchymal cells. These findings support the concept that the hepatocyte has a central role in the uptake and storage of transferrin iron.
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PMID:Heterogeneous distribution of transferrin receptors on parenchymal and nonparenchymal liver cells: biochemical and morphological evidence. 353 27

Dome and dome epithelial cells were selectively dissociated from gut-associated lymphoid tissues of rabbits. Sequential tissue washes in dithiothreitol, EDTA, and collagenase removed the dome epithelium, without disrupting the follicles or villi, and provided a cell suspension containing 74 +/- 6% lymphocytes, 9 +/- 4% columnar epithelial cells, 10 +/- 7% tangible-body macrophages, and 4 +/- 2% M cells (follicle-associated epithelial cells). The last mentioned cells were characterized by transmission electron microscopy as large (20 to 55 microns diameter) cuboidal, round, or oval cells with eccentric nuclei and thin membranous processes surrounding empty vacuoles. The M cells were occasionally joined together by tight junctions. Histochemical and immunocytochemical analyses of M cells with the light microscope showed that they were devoid of immunoglobulins and negative for T-cell antigen and secretory component and had no detectable alkaline phosphatase or endogenous peroxidase activity. The M cells had few vacuoles with faint acid phosphatase activity; nonspecific neutral esterase was abundant. Possible uses for dome and dome epithelial cells are discussed.
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PMID:Dome epithelial M cells dissociated from rabbit gut-associated lymphoid tissues. 354 8

Specific radioimmunoassays for the 7-S domain of type IV collagen and the fragment P1 of laminin were used to quantify these basement membrane proteins in human kidney cortex at different ages and in some patients with diabetes mellitus. The antigens were solubilized by treating the tissue samples with the proteolytic enzymes collagenase, trypsin and pepsin. Total collagen content (as indicated by hydroxyproline concentration) increased with age, and the proportion of the collagen that could be solubilized by any enzyme treatment decreased. The type IV collagen concentration increased significantly with age, whereas the laminin concentration tended to decrease. In the one case of a type I diabetic the amounts of both antigens exceeded those in the age matched controls. In four type II diabetics the results were comparable with those for other aged cases. The distribution of the proteins was studied using the peroxidase-antiperoxidase method. The staining intensity and thickness of both antigens increased with age in the mesangium and Bowmans capsules, the change in type IV collagen staining being more evident. In diabetic patients these changes were more pronounced and other basement membranes appeared thicker in the stainings. These results indicate that basement membrane material accumulates in the kidney cortex during aging and that an alteration takes place in the composition of the basement membranes, the proportion of type IV collagen increasing and that of laminin decreasing.
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PMID:Effect of age and diabetes on type IV collagen and laminin in human kidney cortex. 378 96

Pinocytosis and prostaglandin E2 production are two major functions of the mononuclear phagocyte system. The goal of this study was to compare the pinocytosis of horseradish peroxidase and the prostaglandin E2 production between the hepatic and peritoneal resident macrophages in the mouse. Hepatic resident macrophages were isolated by collagenase digestion, differential centrifugation and adherence. Peritoneal resident macrophages were isolated by peritoneal cavity washing followed by adherence. Horseradish peroxidase was endocytosed by hepatic macrophages at a significantly higher rate (118 +/- 12 ng/10(6) cells/60 min) than by peritoneal macrophages (21 +/- 4 ng/10(6) cells/60 min). Prostaglandin E2 production was measured in the culture medium of unstimulated and lymphokine-stimulated hepatic and peritoneal resident macrophages. Prostaglandin E2 concentration in the culture medium of unstimulated peritoneal macrophages was 36.6 +/- 26.8 ng/ml after a 24 h incubation. It was increased by 83 p. 100 in presence of a lymphokine-enriched secondary mixed lymphocyte culture supernatant. In contrast, hepatic macrophages did not produce any significant amount of prostaglandin E2, even if they were incubated in presence of lymphokines. This study shows that hepatic resident macrophages have a higher pinocytic capacity than peritoneal resident macrophages, suggesting that the role of the liver in the clearance of gut-derived antigens is not only due to its portal irrigation but also to the presence of macrophages highly differentiated in their endocytotic properties. The lack of prostaglandin E2 production in hepatic macrophages, in basal conditions as well as after lymphokine-stimulation, suggests that these cells play a minor role in the regulation of the immune response.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:[Comparative study of pinocytosis and the production of prostaglandin E2 by hepatic and peritoneal resident macrophages in mice]. 386 Apr 55

An immunohistochemical method was used to demonstrate the presence of gonadotrophins in isolated ovarian interstitial cells. The cells were obtained by collagenase digestion of large ovarian follicles after removal of the yolk and the granulosa layer. Using a peroxidase-labelled anti-rabbit serum with anti-chicken follicle stimulating hormone (FSH) serum raised in rabbits, a strong positive reaction was obtained. Anti-human FSH serum also produced a positive result but the reaction was weaker. There was no apparent difference in the staining reaction of cells which had been preincubated with ovine FSH serum. Treatment with anti-ovine luteinizing hormone (LH) resulted in a faintly positive reaction. The viability of the cells was tested by the Trypan Blue method and they were identified as steroid-producing cells by the histochemical demonstration of their 3 beta-hydroxysteroid dehydrogenase activity.
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PMID:Detection of gonadotrophic hormones on isolated thecal interstitial cells of the domestic hen, Gallus domesticus, by an immunohistochemical method. 388 6

Although some data suggest that macrophages in the reticuloendothelial system (RES) are important sources of thromboxane A2 (TxA2) and prostacyclin (PGI2) during endotoxic shock, we are unaware of data documenting the ability of hepatic macrophages (Kupffer cells) to release either TxA2 or PGI2 when exposed to lipopolysaccharide (endotoxin, LPS). In this study, Kupffer cells were examined for their ability to release prostaglandin E2 (PGE2), TxA2, and PGI2 following stimulation with 0, 1.0, 50.0, and 100.0 micrograms/ml of Escherichia coli LPS. Kupffer cells were obtained from rat livers by enzymatic digestion with 0.05% collagenase followed by enrichment of the macrophage population on the basis of differences in density and adherence among the various cell populations isolated. Based on several criteria (phagocytosis of opsonized sheep erythrocytes, positive staining for esterase and peroxidase, failure to replicate), 95% of adherent cells were Kupffer cells. After 4 days of incubation, cells were stimulated with various doses of LPS for 4 and 8 hr. Prostanoid concentrations in culture supernatants were determined by radioimmunoassay. Increasing doses of LPS significantly (P less than 0.001) increased the concentration of immunoreactive PGE2 (iPGE2) and iTxB2 (the stable metabolite of TxA2). The concentration of i6-keto-PFG1 alpha (stable metabolite of PGI2) increased following stimulation with 1.0 microgram/ml of LPS, but declined as the dose of LPS was increased. The results provide evidence that endotoxin-activated Kupffer cells, like other macrophage populations, release several metabolites of arachidonic acid. Kupffer cell-derived prostanoids, particularly TxA2, may be important mediators of some of the pathophysiologic manifestations of acute endotoxemia.
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PMID:Prostanoid production by lipopolysaccharide-stimulated Kupffer cells. 388 37

Antigen-retaining follicular dendritic cells (FDC) have been identified and studied in sections of lymph nodes and spleen, but studies of these cells in culture have been extremely limited. The purpose of this study was to establish techniques to release these fragile cells from mouse lymph nodes in a viable state and to identify these cells routinely in lymph node cell suspensions. FDC were obtained from passively or actively immunized popliteal lymph nodes of mice injected in the footpads with 125I-labeled human serum albumin (HSA) or horseradish peroxidase (HRP). Lymph nodes were removed 1 hr after the footpads had been injected with collagenase. After another hour of incubation in vitro with collagenase, protease, and deoxyribonuclease, FDC were released by gentle teasing and enriched by centrifugation on a low density bovine serum albumin (BSA) or Percoll gradient. Most FDC with the associated radiolabel floated at densities greater than 1.06 g/ml on BSA or Percoll gradients. Slides of the FDC-enriched fraction were prepared, using a cytobucket which allowed the cells to be affixed to glass slides by centrifugation in a less disruptive manner than by cytocentrifugation. FDC that were air-dried and fixed with 3% glutaraldehyde had a characteristic pink acidophilic cytoplasm after Wright's staining, and had a faintly basophilic euchromatic nucleus frequently with peripherally-clumped chromatin. In addition, these cells were large and irregularly shaped (up to 60 micron long). Fixation of FDC with 0.6% paraformaldehyde/ 0.9% glutaraldehyde on poly-L-lysine-coated slides resulted in a preservation of FDC which made possible visualization of long dendritic processes by Nomarski optics. Antigen presence on the cell surface was confirmed by autoradiography and, in the case of HRP, was also visualized enzymatically using diaminobenzidine. In contrast to resident peritoneal macrophages or some contaminating lymph node macrophages present on the same slides, FDC did not phagocytize opsonized sheep red blood cells (SRBC) or adhere to plastic surfaces although they did form rosettes with opsonized SRBC. Cell marker studies indicated FDC have a distinctive phenotype. They were positive for Ia, Fc receptor, and leukocyte common antigen, but negative for Thy-1, Ly-1, Ly-2, endogenous Ig, Mac-1, Mac-2, Mac-3, and F4/80, and negative to weakly positive for nonspecific esterase. Cultured FDC remained viable and retained radiolabeled antigen-antibody complexes on their surfaces and were significantly enriched for FDC.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Follicular dendritic cells in suspension: identification, enrichment, and initial characterization indicating immune complex trapping and lack of adherence and phagocytic activity. 396 23


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