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)

The process of connective tissue breakdown in chronic otitis media is described in the context of recent advances in our understanding of collagen degradation and bone resorption. The significance of the initial step in collagen breakdown, brought about by the action of a specific collagen dissolving enzyme is emphasized in terms of recent studies in other chronic inflammatory diseases characterized by connective tissue breakdown. Bone resorption, a characteristic feature of chronic otitis media, requires the breakdown of collagen, which comprises over 90 percent of bone protein. Evidence in support of collagenase in bone resorption from adjacent tissue (in this case, inflammatory connective tissue) would require identification of the enzyme in cells involved in the inflammatory process adjacent to the resorbing bone. Collagenase was found localized in frozen sections of canal wall skin, middle ear granulation and in cholesteatoma by a specific binding of the enzyme with an antiserum produced against purified human skin collagenase. The antigen antibody complex was labelled with fluorescein. Collagenase appeared in the subepithelial connective tissue of cholesteatoma, granulation tissue from the middle ear and the dermis of canal skin; but was not seen in the keratin layer, epithelium or the epidermal appendages. The enzyme appeared within certain fibroblasts, macrophages and endothelial cells of capillary buds. Collagenase enhanced by chronic inflammation attacks the intact collagen molecule, making it susceptible to further digestion by other proteases that are also products of inflammation. This process brings about resorption of connective tissue and bone.
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PMID:Localization of collagenase in human middle ear cholesteatoma. 19 5

The composition of extracellular proteinases from Actinomyces fradiae 0072 and their effect on proteins and synthetic substrates were studied. The enzyme preparation was found to have keratinolytic, caseinolytic, collagenolytic, collagenase, trypsin-like, carboxy- and aminopeptidase activities. Five low molecular weight proteinases capable to hydrolyse keratin, casein, azocollagen were obtained via fractionation of the enzyme preparation on DEAE-cellulose and Sephadex columns. Proteinases from Act. fradiae and aminopeptidase with a molecular weight of 31,000 were shown to be different enzymes. In hydrolysis of L-leucyl-2-naphthylamide the Michaelis constant and Vmax of the enzyme were found to be 3.02 X 10(-3) M and 0.35 X 10(2) micronM/ml-min, respectively.
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PMID:[Characteristics of extracellular proteolytic enzymes of Actinomyces fradiae 0072]. 19 35

The antibodies of keratin and vimentin were used as the histochemical probes determined by the immuno-fluorescent technique to recognize the rat epididymal epithelial cells in the different ages from the connective tissue both in intact epididymides and in isolated cultured cells. It also showed that an enriched suspension of epididymal epithelial cells could be obtained by sequential digestion with 0.05% trypsin and 0.1% collagenase. The morphological characteristics were appeared during the cells in culture. Therefore the epididymal epithelial cells isolated and cultured by present methods could be used as a research model to study their functions.
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PMID:[The recognition of rat epididymal epithelial cells]. 170 22

A protocol which was developed for the culture of epithelial cells from radical prostatectomy specimens was slightly modified to permit the culture of cells from ultrasound-guided prostatic needle biopsies. The collagenase digestion step of the standard protocol was omitted, and biopsies were simply minced and allowed to attach to collagen-coated dishes in serum-free medium. Cell outgrowths from biopsies were free of fibroblasts, and expression of keratin, prostate specific antigen, and prostatic acid phosphatase was maintained in vitro. The establishment of a bank of frozen cells from primary cultures permitted repetitive studies with individual cell strains, which could be serially passaged and were capable of clonal growth. The ability to derive cultures from biopsies will facilitate the biological characterization of cells from primary prostate tumors of high malignant grade, which are not commonly available from radical prostatectomy specimens.
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PMID:Culture of prostatic epithelial cells from ultrasound-guided needle biopsies. 171 64

Methacholine (MCh)- and isoproterenol (Iso)-stimulated 14CO2 production was compared between freshly dissociated rhesus sweat secretory coil cells (mainly clear cells) and cultured cells (grown on a collagen-coated plastic plate) derived from native cells. 14CO2 production was enhanced by MCh and by Iso in native coil cells (but not in ductal cells) in a pharmacologically specific and dose-dependent manner. 14CO2 production in subcultured coil cells (19-45 days in culture) was only one-third to one-fifth that of native cells. MCh-stimulated 14CO2 production was inhibited by ouabain and furosemide in both native and cultured coil cells. A decrease in 14CO2 production, of about one-half, was already evident in primary cells cultured for less than 1 wk. The decreased pharmacological responsiveness of the cultured coil cells was seen, although the cultured cells showed the typical epithelioid appearance, abundant mitochondria, the occasional presence of intercellular lacunae resembling intercellular canaliculi, and the persistence of immunoreactive keratin. We conclude that 1) a primary culture of sweat gland cells can be initiated from dissociated cells; 2) cultured sweat secretory coil cells qualitatively, but not quantitatively, retain the pharmacological responsiveness and transport activity of the native cells as determined by 14CO2 production; 3) collagenase-dissociated cells represent an excellent in vitro system for the study of glandular function at the cellular level; and 4) the decrease in pharmacological responsiveness is not simply due to trypsin treatment during harvesting of cultured cells, because that of organ-cultured, intact, secretory coils also declines with time of culture.
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PMID:Pharmacological responsiveness of dissociated native and cultured eccrine secretory coil cells. 236 Jun 86

The connective tissue adjacent to basal cell carcinomas (BCC) is frequently abnormal and contains increased numbers of fibroblasts and increased extractable collagenase. To determine whether BCC could produce these alterations by releasing mediators that regulated fibroblast function, we established BCC in culture and tested the ability of their culture supernatants to alter fibroblast proliferation and production of collagenase. Using tissue culture plates coated with type IV collagen and containing x-irradiated 3T3 feeder cells, we established epithelial colonies from 47% of the BCC cultured. The BCC-derived colonies differed from normal epidermal cell colonies in their morphology, growth rate, and keratin production. Culture supernatants from 4 out of 5 confluent BCC-derived colonies contained factors that stimulated fibroblasts to proliferate and release collagenase. These findings show that BCC-derived epidermal cell colonies release mediators which alter fibroblast functions and suggest that some of the connective tissue changes associated with BCC in vivo are the result of BCC-fibroblast interactions.
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PMID:Establishment of basal cell carcinoma in culture: evidence for a basal cell carcinoma-derived factor(s) which stimulates fibroblasts to proliferate and release collagenase. 241 70

Epididymal epithelial fragments, free of stromal elements were isolated from mature rats using two sequential collagenase digestions. Within 24 h these attached efficiently to a variety of substrates including glass, plastic, placental collagen, type IV collagen and epididymal extracellular matrix material. Cells spreading away from the fragments rapidly assumed a flattened, overlapping, monolayer appearance typical of epithelial cells in culture. Cells still associated with the fragments or adjacent to them remained more polarized and more closely resembled epididymal principal cells in vivo than did cells that had migrated to the periphery of the monolayer. Apical microvilli characteristic of these cells in vivo were common during the first 4 days in culture but diminished in number and size thereafter. Cultured cells maintained many of the structural features characteristic of principal cells in vivo, including a well developed Golgi apparatus, coated pits and vesicles, and many multivesicular bodies. An extensive filamentous network, shown immunocytochemically to consist of keratin, was present in the cytoplasm of all cells but was more obvious in flattened cells at the periphery of the monolayer. Rhodamine phalloidin labelling of filamentous actin showed that concentrations of actin occurred corresponding to microvilli on the apical surface, in a continuous ring just below the apical surface, and also in stress fibres at the base of the cells. Cells isolated and cultured from the distal caput epididymidis possessed lobulated nuclei, in contrast to the round or oval nuclei found in cells cultured from the proximal caput epididymidis. Cells from the distal caput epididymidis were also characterized by the presence of many lipid droplets in their cytoplasm. Autofluorescent granules were observed in epithelial cells from both regions but were larger and more numerous in cells isolated from the distal caput epididymidis. Tritiated thymidine incorporation by the cells after 4 days in culture showed that cells adjacent to the parent epithelial fragment were dividing at a greater rate than cells that had migrated to the periphery of the monolayer.
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PMID:Structural features of rat epididymal epithelial cells in vitro. 241 3

A culture of cells derived from the endometrium was established to study in vitro the effects of hormone on the endometrium. Endometrial tissue was cut into small pieces and suspended in trypsin and collagenase-containing medium. The primary cell culture was obtained by six passages. The following observations appeared to indicate that the cultured cells consisted of endometrial epithelium-derived adenocytes, endometrial stromal cells and fibroblasts: 1. The epithelium-derived adenocytes and the endometrial stromal cells were positive and the fibroblasts were negative for staining with keratin by the enzyme-labelled antibody method. 2. Transmission electron microscopic observation revealed microvilli on the surface of the adenocytes, abundant free ribosomes in the cytoplasm, and nucleoli in distinct nuclei whose margin was deeply stained. Junction complexes among the cells were also observed. There were no microvilli on the surface of the endometrial stromal cell. Filaments, mitochondria and secreting granules were distinctly seen in the cytoplasm. The adenocytes on the margin of the nucleus itself were more deeply stained, and the margin was notched. The fibroblasts were spindle-shaped, and there no basement membrane structure laterally along the surface of the fibroblast. There were no intercellular junction complexes but there were filaments in the intercellular borders and in the cytoplasm. Chromatin in the large oval nuclei was dispersed.
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PMID:[Establishment of primary culture of cells derived from human uterine endometrium]. 246 25

Enriched epithelial cell and fibroblast fractions were isolated from ovine placentomes by isopycnic centrifugation of collagenase/DNAse-dispersed cells through a density gradient of 45% Percoll. The epithelial cells formed confluent monolayers when plated onto filters impregnated with a 50-microns layer of Matrigel in medium containing 10% fetal bovine serum. These cells were maintained in dual environment culture chambers in serum-free medium for at least 12 days. The epithelium had a polarized appearance similar to that found in vivo only when cells were plated at high density (10(7)/cells/cm2). The epithelial monolayer consisted predominantly of a single population of uninucleate cells with intracellular features similar to those previously described for ovine trophoblast both in vivo and in vitro. These cells stained positively with an antiserum to alpha-keratin, a marker specific to epithelial cells, and no staining was observed with antisera raised against binucleate cells or leucocyte-common antigen. Binucleate cells were detected by microscopy and immunostaining in the pellet of cells obtained from the Percoll gradient but were rarely seen in the epithelium. The epithelial monolayer excluded 3H-inulin, added to the basal chamber, from the apical chamber, thus demonstrating the formation of a permeability barrier similar to that found in vivo. The maintenance of a monolayer of pure ovine trophoblast cells in vitro, which retain the characteristics of the epithelium in vivo, will enable the study of many cellular functions of the trophoblast.
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PMID:Identification and characterization of monolayer cultures of sheep trophoblast cells maintained in bicameral culture chambers. 248 Jan 61

Parenchymal organoidal structures that were obtained from collagenase digestion of reduction mammoplasty specimens of apparently normal human breasts have been grown in short-term primary cultures, either on plastic or on floating gels of polymerized rat-tail collagen. Three morphologically distinct major cell types are readily observed in both systems: cuboidal cells, which occupy apical positions on collagen gels; larger, epithelioid, or basal cells on gels; and elongated cells which penetrate into the gel. In addition, a fourth cell type, that of large, flat cell, is observed less readily by phase contrast microscopy on the surface of cultures grown on plastic. Immunofluorescent and immunocytochemical staining of cultures on plastic or histologic sections of cultures on gels have been undertaken with antisera and other histochemical reagents that stain the different parenchymal cell types in vivo. Thus antisera to epithelial membrane antigen(s), monoclonal antibodies (MABs) to the defatted mammary milk fat globule membrane, peanut lectin, and keratin MAB LE61, which preferentially stain the epithelial cells of ducts in vivo, also stain the cuboidal/apical cells in vitro. The large, flat cells are stained intensely by the first three reagents but not by the last one. Antisera to collagen IV, laminin, fibronectin, actin, keratin MAB LP34, MABs to the common acute lymphoblastic leukemia antigen, and MAB LICR-LON-23.10, which showed enhanced staining for the ductal myoepithelial cells in vivo, also stain the epithelioid/elongated cells in vitro. However, the effect of the last four reagents is reduced considerably in most elongated cells, and MAB LP34 stains the large, flat cells intensely. Heterogeneous cells of intermediate morphologies and staining patterns between the cuboidal cells and large, flat cells are related to mammary epithelial cells. whereas the large epithelioid/elongated cells have some characteristics of myoepithelial cells, and that intermediate forms may exist in culture between the two parenchymal cell types.
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PMID:Characterization of human mammary cell types in primary culture: immunofluorescent and immunocytochemical indicators of cellular heterogeneity. 264 83


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