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 objectives of this study were to establish a suitable and validated in vitro bioassay of piscine gonadotropins (GTHs) by using a carp testis androgen production system and to compare the androgenic responses in such an assay to gonadotropins from various vertebrate species. The testes from mature carp with gonadosomatic indices of 8-30% were used. Androgen production was first compared with respect to methods for preparation of the carp testis (sliced, minced, homogenized, and collagenase-dispersed testis preparations). The time course of androgen formation, the effects of xanthine and theophylline, and other factors on androgen production also were investigated. Theophylline was more effective than xanthine in potentiation of gonadotropin-evoked androgen formation by carp testis. The testis preparations were incubated in medium 199 (pH 7.40) containing 2 mM theophylline with shaking at 100 cycles/min at 25 degrees C for 4 hr. Homogenized testis preparations had limited ability for androgen production, while sliced, minced, and minced-collagenase-dispersed testis preparations were highly responsive to gonadotropins for androgen production. The minced testis preparation, utilizing 100 mg/ml incubation medium per vial, was chosen as the standard incubation procedure in this study. The minced testis androgen production assay was highly sensitive to gonadotropins from several piscine species (silver carp, common carp, and salmon), and all these GTHs produced parallel dose-related androgen production curves. Mammalian GTHs were also capable of promoting androgen formation by carp testis, but they were much less potent than were piscine GTHs. Pregnant mares' serum gonadotropin (PMSG) was more effective than human chorionic gonadotropin (hCG) in evoking carp testis androgen production.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Species variation of gonadotropic activity in an in vitro assay measuring androgen formation by carp (Cyprinus carpio) testis with special reference to bioassay of piscine gonadotropins. 377 Apr 34

Surfactant-associated glycoprotein A [molecular weight (Mr) = 34,000, isoelectric point (pI) 4.6-5.0] and its sulfhydryl dependent oligomers were purified and partially characterized from surfactant obtained from human alveolar lavage. Two major forms of the protein were identified by silver stain and immunoblot analysis of surfactant using human surfactant-associated glycoprotein A antisera: glycoprotein A2, Mr = 34,000 and glycoprotein A1, Mr = 28,000. The larger form was reduced to Mr = 28,000 by treatment with endoglycosidase F, indicating the presence of complex N-linked oligosaccharide on the molecule. Charge heterogeneity was decreased and the isoelectric point increased by treatment with neuroaminidase, supporting the presence of sialic acid. Homology between the proteins Mr = 34,000 and 28,000 was confirmed by analysis of two-dimensional tryptic and chymotryptic peptides of 125I-iodo-glycoproteins A1 and A2 which were identical. The protein was very rich in glycine and its amino acid composition was similar to that of glycoprotein A previously reported for the dog and rat. Treatment of glycoproteins A with bacterial collagenase resulted in the generation of highly glycosylated peptides Mr = 20,000-22,000, pI 4.6-5.0, which no longer formed sulfhydryl-dependent oligomers, supporting the presence of significant collagen-like region in the molecule. In the absence of reducing agents, glycoprotein A from surfactant was present as sulfhydryl-dependent dimers and larger oligomers. Higher molecular weight aggregates of glycoproteins A were also present in lavage material even after sulfhydryl reduction. Glycoproteins A were identified in surfactant from amniotic fluid, normal adult lung lavage, human cadaver lung lavage, and material obtained from lung lavage from a patient with alveolar proteinosis.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Characteristics of human surfactant-associated glycoproteins A. 383 68

Neuroepithelial bodies (NEB) in 29-day fetal rabbit lung were examined by light microscopy and cytochemistry to demonstrate their structural and biochemical properties in situ. Longitudinal sections of NEB at airway bifurcations demonstrated their chemoreceptor-like appearance. Furthermore, the cytochemical presence of serotonin, acetylcholinesterase, formaldehyde-induced fluorescence, and silver-staining properties demonstrated the neural-like biochemical properties of NEB cells. Forty-one NEB and eight single neuroendocrine cells from whole fetal lungs were examined ultrastructurally. Juxtaluminal junctional complexes composed of tight and intermediate junctions, desmosomes, and cytoplasmic filaments were demonstrated in the corpuscular-shaped NEB. Basal bodies were apparent in NEB cell cytoplasm; cilia extended from NEB cells. Dense-core vesicles (DCV) were of at least three types: type 1, type 2, and enterochromaffin type. The majority of epithelial cells adjacent to NEB in near-term airway epithelium were undifferentiated, with large amounts of glycogen. However, ciliated cells were adjacent to some small NEB and single neuroendocrine cells; mucus or Clara-type cells were not observed. NEB isolated by collagenase treatment revealed an intact organoid structure, DCV, and desmosomes and retained their argyrophilia and formaldehyde-induced fluorescence. NEB were recovered in cell fractions separated by unit gravity that had cells in clumps of four or more. One to five NEB stained with silver in cytocentrifuge preparations of control, mixed cells, whereas up to 20 intact NEB were demonstrated in the clump-containing, separated fractions. We propose that isolated NEB retain certain biochemical and metabolic properties similar to those of their counterparts in situ. Serotonin and 5-hydroxyindole acetic acid were found by high-performance liquid chromatography analysis in the fractions containing NEB, and amine precursor uptake and decarboxylation (APUD) activity were demonstrated. Moreover, muscarinic cholinergic receptors were detected, consistent with the occurrence of acetylcholinesterase in NEB. The elution profile of bombesin radioimmunoactivity substantiated that isolated fetal rabbit NEB contained this neuropeptide and that NEB were enriched by unit gravity sedimentation. These studies suggest that NEB are structurally and functionally developed before other cell types in immature airway epithelium and can be isolated as intact organoids, which retain some of their structural and metabolic integrity.
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PMID:Morphological and cytochemical characterization of neuroepithelial bodies in fetal rabbit lung. I. Studies of isolated neuroepithelial bodies. 613 13

Whole retinae of 4- to 10-day-old chick and quail embryos were spread on membrane filters and kept in culture for up to 4 days. Axon growth during culture was demonstrated by silver staining, anterograde labeling of fibers with RITC, time-lapse recording, and SEM. Fiber growth was observed in specimens from chick embryos up to 7 days old, with a growth maximum at E6 and from quail embryos up to E6 with the maximum at E5. Newly growing axons followed the optic fiber pattern already existing and, like axons in vivo, grew predominantly toward the optic fissure. Directional and orientational adaptation of newly growing axons to the preexisting fibers increased with the donor age. Retinae from donors up to E5 in chick and up to E4 in quail showed a high proportion of axons which crossed the optic fissure during the culture period and invaded the opposite retinal fiber layer. These fibers showed a correct radial orientation while growing in the opposite direction to normal. Likewise, in cultures from these young donors some fibers grew out initially in the diametrically opposite direction to normal toward the tissue periphery. Since all of the wrongly directed axons grew at the same rate as normal and adapted correctly to the already formed axon pattern, this suggests independent signals for the direction and orientation of growing fibers. Treatment of mounted retinae with collagenase or trypsin removed the vitreal retinal surface, leaving the existing axon pattern intact. Subsequently, new axons grew profusely in culture, but lost both their orientational and directional characteristics.
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PMID:Axon growth in embryonic chick and quail retinal whole mounts in vitro. 620 Mar 72

The sites of action of aldosterone (A) along the tubule of rabbit kidney were studied by autoradiographic localization of mineralocorticoid-binding sites on microdissected tubular segments. Kidney pyramids were incubated at 30 degrees C for 1 h in a collagenase solution with [3H]aldosterone at a concentration of 1.5 X 10(-9) M with and without an excess unlabeled A. Tubular segments were then microdissected and transferred onto dry film; fixation and staining were done only after exposure of the film 4 mo later in order to avoid diffusion. Specific nuclear labeling was 19.0 +/- 1.3 silver grains/100 micrometers2 in distal convoluted tubules (n = 28) and 21.0 +/- 1.8 in cortical collecting ducts (n = 18). No difference between these two structures was observed (P greater than 0.1, paired t test, n = 15). No specific binding was found in the proximal tubule (0.5 +/- 0.4, n = 17). In the thick ascending limb of Henle's loop, the labeling was low (3.9 +/- 0.9, n = 16). We conclude that, in the rabbit kidney, nuclear mineralocorticoid-binding sites, presumably receptors, are present in the distal and cortical collecting tubule.
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PMID:Aldosterone binding along the rabbit nephron: an autoradiographic study on isolated tubules. 625 52

Thyrocytes were isolated from porcine thyroid glands using a new method entailing two-step collagenase digestion and Percoll gradient centrifugation. Good yield and a high percentage of viable thyrocytes free from other contaminating cells was achieved. Proteins present on the surface of thyroid cell plasma membranes were then specifically labeled using Iodogen and 125I-. Membrane lysates were separated by electrophoresis on 10% polyacrylamide gel, under reducing and nonreducing conditions, followed by autoradiography. When the gels were stained with silver nitrate some 30 bands were visualized in both the presence and absence of reductant. Only 9 bands were found to be labeled under nonreducing conditions and 12 in the presence of reductant. Two bands involved in the thyrotropin receptor structure--Mr = 66,000 and Mr = 70,000, respectively--were visualized in the absence of reductant. Upon reduction the Mr 66,000 band was retained and a new band (Mr = 33,000) was seen. The mild enzymatic treatment used in isolating thyrocytes and the lack of contamination with other cells allowed the consistent labeling of exposed plasma membrane components by the Iodogen method such that the orientation of thyrotropin receptor components in the plasma membrane could be deduced.
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PMID:Surface labeling of thyrocytes isolated by a new method combining enzymatic digestion and percoll gradient centrifugation. 630 79

We have investigated the effects of the tumor-promoting phorbol diester, 12-O-tetradecanoylphorbol-13-acetate (TPA), on rabbit synovial fibroblasts, and found that this agent induced a major switch in gene expression in these cells that was marked by the specific induction of the neutral proteinase, collagenase, and was always accompanied by alterations in cell morphology. Procollagenase synthesis and secretion was first observed 6-12 h after the addition of TPA. The rate of collagenase production (1-5 U, or approximately 0.2-1 micrograms secreted procollagenase protein per 10(5) cells per 24 h) depended on the TPA concentration (1-400 ng/ml) and time of exposure (1-72 h). Procollagenase was the most prominent protein visible by direct silver staining or by autoradiography after SDS PAGE of [35S]methionine-labeled proteins. The two procollagenase bands of Mr 53,000 and 57,000, which migrated as a family of spots on two-dimensional gels and were immunoprecipitated by antibodies to purified rabbit collagenase, accounted for 23% of the newly synthesized, secreted protein in TPA-treated cells. Cell-free translation of mRNA from TPA-treated cells in rabbit reticulocyte lysate produced a single band of immunoprecipitable preprocollagenase (Mr 55,000) as a major product (5% of total) that migrated as a single spot on two-dimensional gels. Secreted procollagenase, preprocollagenase , and active collagenase (purified to homogeneity; specific activity 1.2 X 10(4) U/mg protein) had related peptide maps. Two other major secreted proteins, a neutral metalloproteinase of Mr 51,000 and a polypeptide of Mr 47,000, were also induced by TPA. In contrast to the induction of these four polypeptides, TPA decreased synthesis and secretion of a number of proteins, including collagen and fibronectin. Thus, collagenase is a convenient marker for major alterations in the pattern of protein synthesis and secretion by rabbit synovial fibroblasts treated with TPA.
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PMID:Collagenase is a major gene product of induced rabbit synovial fibroblasts. 632 17

The presence and the ontogeny of class I and class II major histocompatibility (MHC) antigens were investigated on human trophoblast cells in single cell suspensions of collagenase dispersed placentae during the first trimester of pregnancy by using a highly sensitive radioautographic approach. Cells were treated with monoclonal anti-HLA-A,B,C or anti-HLA-DR (or anti-human Ia) antibodies followed by 125I-labeled protein A, normal mouse serum, or medium alone used as negative controls, and monoclonal antibodies against trophoblast specific antigenic markers (anti-Trop 2, NDOG-1, and NDOG-2) used as positive controls for specificity of trophoblast labeling. Tissue controls were provided by Ficoll-paque-separated adult human peripheral blood lymphocytes (PBL), and placental lymphocytes were used as additional internal controls. Results revealed variable but specific labeling of cytotrophoblast cells for class I but not class II MHC antigens at all gestational stages during the first trimester. The incidence of HLA-A,B,C antigen-bearing cytotrophoblast cells in early gestational (5 to 8 wk) placentae was high (75 to 91%) in the minority, but was low to medium (35 to 66%) in placentae at later gestational intervals (9 to 12 wk). The initial antigen density on the labeled cytotrophoblast cells as reflected by the median silver grain counts per unit area was approximately 75% of that noted on adult PBL or placental lymphocytes at 5 to 7 wk. The density then dropped to 50% of the initial level by the eighth week of gestation with no further change up to 12 wk. Syncytiotrophoblast cells revealed no appreciable labeling for either antigen class. This could not be explained by antigen stripping due to the collagenase dispersion procedure; specific labeling was not detectable on the syncytiotrophoblast cell layer in situ in histologic sections after intact chorionic villi had been directly exposed to 125I-labeled monoclonal anti-class I antibody. These results are qualitatively similar to our findings in the mouse in revealing the presence of class I MHC antigens on a major population of trophoblast cells.
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PMID:Ontogeny of the MHC antigens on human trophoblast cells during the first trimester of pregnancy. 641 63

The growth, synthesis and regional specialization of the lens capsule has been studied in chicken embryos and compared to adult chickens and mammals. During the final 15 days of embryonic development the surface area of the capsule increased 11-fold. This represents the minimum estimate of capsule growth, since it also increased in thickness during this period. Autoradiographic examination of [3H]proline- or 35SO4-labeled, detergent-cleaned, flat-mounted capsules suggested that all lens cells participated in capsule synthesis. This was supported by the observation that both lens epithelial and fiber cells incorporated [35S]-methionine into collagenase-sensitive proteins with molecular weights similar to type IV collagen. Histochemical staining of detergent-cleaned capsule explants (DCCEs) revealed regional differences in the carbohydrate composition of the capsule. Differences in collagenous proteins were also seen between the anterior and posterior regions of the capsule of the embryonic lenses. Following SDS-PAGE and silver staining a total of six collagenase-sensitive proteins were seen with molecular weights between 150 and 180 K. Three of these polypeptides were common to both anterior and posterior capsules, one was found only in the anterior capsule and the other two were found predominantly in the posterior capsule. No regional differences were seen in the collagenous proteins of capsules from adult chickens, rats or hamsters. The possibility is discussed that the unique pattern of collagenous polypeptides seen in embryonic capsules is related to the rapid growth of the embryonic lens.
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PMID:Growth, synthesis and regional specialization of the embryonic chicken lens capsule. 646 38

The dose-dependent effects of heavy metals on cell proliferation, collagen synthesis, and non-collagen protein synthesis were studied in early passage cultures of human synovial cells exposed to 1-100 microM concentration of gold, silver, mercury, cadmium or lead for 5 days. The incorporation of [3H]thymidine into trichloroacetic acid insoluble material was inhibited 50% by each of the heavy metals at concentrations between 1 and 10 microM. Gold, lead and mercury (10 microM) decreased the DNA content of the cultures by less than 15%; silver (10 microM) and cadmium (10 microM) resulted in decreased DNA content, which was attributed to cytotoxicity. A dose-dependent inhibition of [3H]proline incorporation into bacterial collagenase resistant (non-collagen) protein was observed after incubation with 10 microM mercury, lead and silver. During incubations with 10 microM gold and cadmium, collagenase resistant protein accumulation increased. All the heavy metals except for gold inhibited collagen accumulation to a greater extent than non-collagen protein accumulation. Gold (10 microM) stimulated the amount of collagen produced per cell, and the percentage of collagen to total protein was increased 50%. The rate of collagen accumulation in medium decreased during incubation with 10 microM silver, mercury, cadmium and lead. The stimulation of collagen synthesis may be a unique property of gold related to the therapeutic indices of gold, compared to other heavy metals, in rheumatoid arthritis.
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PMID:Effect of heavy metals on human rheumatoid synovial cell proliferation and collagen synthesis. 662 46


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