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)

Effect of diltiazem on glucose-induced insulin secretion was investigated in the rat islets of Langerhans isolated by a collagenase digestion technique. It was found that B-cells, main constituents of isolated islet preparations, had a well-preserved ultrastructural appearance immediately following isolation or after incubation with glucose or glucose and diltiazem. The islets released a large amount of insulin upon stimulation with glucose and CaCl2. Diltiazem (10(-6)-10(-4) M) produced a dose-related inhibition of glucose-induced insulin secretion and this effect was antagonized by the increase in extracellular concentration of CaCl2. The inhibitory effect of diltiazem on the insulin secretion was also counteracted by dibutyryl-3',5'-cyclic AMP or by theophylline. Among calcium-antagonists tested, nifedipine produced the most powerful inhibitory action on the insulin secretion, while the effect of verapamil was similar to or somewhat stronger than that of diltiazem. It was suggested that diltiazem may reduce the intracellular concentration of free calcium ion, thus causing an inhibitory effect on the glucose-induced insulin secretion by the isolated islets of Langerhans.
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PMID:Effect of diltiazem on insulin secretion. I. Experiments in vitro. 20 92

Collagenase, a proteolytic enzyme, was injected intradiscally in nine clinically normal, middle-aged beagles. Calcium chloride diluent solution (control), 100 ABC units of collagenase, and 250 ABC units of collagenase, were injected in randomly selected intervertebral discs (T13-L1 to L5-L6). On day 11, the discs injected with collagenase were narrowed radiographically, but there was no significant change in myelograms. Grossly and histologically, there was dissolution of the intervertebral discs, mainly nucleus pulposus, and protrusion of nucleus material in the vertebral body through bony end-plates in discs injected with collagenase. Collagenase chemonucleolysis may be an alternative to spinal surgery for intervertebral disc protrusion in dogs.
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PMID:Chemonucleolysis with collagenase. A radiographic and pathologic study in dogs. 132 Jul 89

Transamidases are a class of calcium-dependent mammalian enzymes which cross-link proteins by catalyzing the formation of (gamma-glutamyl)-epsilon-lysine bonds. It is possible that these enzymes play an important anabolic role in tissue healing. This study was to quantitate transamidase activity in human gingival tissue and examine the relation between transmidase activity and degree of inflammation. Forty-four out of a total 120 collected human gingival specimens from healthy and diseased patients were selected based on histometric and microbiologic criteria. Specimens were minced and homogenized in 10 mM CaCl2 and then extracted for 30 min, in 50 mM tris-HCl buffer (pH 7.5) containing 100 mM CaCl2. Following low speed centrifugation at 4 degrees C, the supernatant solution was assayed for both transamidase and collagenase activities by radioactive amine incorporation, and digestion of tritiated collagen, respectively. Appreciable levels of transamidase and collagenase activities in healthy gingivae were found. These enzyme activities were significantly elevated in the diseased and healing tissues. Unlike other transamidases, calcium was required in the enzyme extraction process.
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PMID:Transamidase and collagenase activity in healthy and diseased human gingival tissues. 146 May 85

The ability of low-dose tetracyclines to inhibit collagenase activity and inactivate osteoclasts suggests that these compounds have great potential as a prophylaxis for metabolic bone disease. However, the cellular mechanism by which tetracyclines interact with skeletal tissue is not yet clear. To better understand the effects of tetracyclines on bone metabolism, we examined their effect on osteoclast activity in vitro. Because tetracyclines can enter the cell and bind calcium and have been reported to directly interact with osteoclasts, we postulated that exposure to either of two tetracyclines, minocycline or doxycycline, would alter cytosolic Ca2+ regulation in rat osteoclasts. [Ca2+]i was measured in single rat osteoclasts utilizing fura-2. Addition of extracellular Ca2+ (5 mM CaCl2), a potent osteoclast inhibitor, increased [Ca2+]i in all osteoclasts, but 10(-6) M salmon calcitonin (sCT) did so only in a subpopulation of osteoclasts. Neither minocycline nor doxycycline (10 micrograms/ml) altered steady-state osteoclast [Ca2+]i. Further, neither minocycline nor doxycycline pretreatment affected the sCT-mediated increases in [Ca2+]i. However, tetracycline pretreatment significantly decreased the cytosolic Ca2+ response to extracellular CaCl2. Our results strongly suggest that tetracyclines have a specific effect on extracellular Ca(2+)-stimulated cytosolic Ca2+ mobilization in osteoclasts, which is not solely dependent on their ability to buffer Ca2+. Furthermore, these results point to the potential use of tetracyclines as probes to study cytosolic Ca2+ regulation. However, that tetracyclines attenuate a signal response associated with decreased osteoclastic resorption suggests that the reported antiresorptive attributes of tetracyclines must be achieved independently of an effect on osteoclastic cytosolic Ca2+.
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PMID:Regulation of cytoplasmic calcium concentration in tetracycline-treated osteoclasts. 146 56

Collagenase is believed to be important for cell migration and collagen remodeling during tissue repair and regeneration. We have investigated collagenase concentrations in different types of surgically inflicted wounds in pigs. Collagenase was extracted from tissue homogenates of wounds by heating to 60 degrees C for 6 min in 0.1 M CaCl2. The molecular weight of latent collagenase was about 52 kDa. Activated collagenase produced the characteristic 3/4 fragment of collagen. Collagenase was assayed by the use of radiolabeled telopeptide-free collagen. To detect maximal collagenase activity, extracts were reduced and alkylated to destroy inhibitors, then activated with aminophenylmercuric acetate. Sutured incisions showed peak collagenase content on postoperative day 1 and thereafter steadily declining concentrations. Granulation tissue from non-sutured large defect full-thickness wounds showed high collagenase content on postoperative day 5 and then a sharp decline to day 7 followed by a slowly declining curve to postoperative day 21. Partial-thickness wounds exhibited a different time course, with collagenase increasing to peak concentrations on postoperative days 3-5; however, a large proportion of the detected collagenase was due to the adherent scab. By day 7 collagenase concentrations approached the low concentrations of normal skin when epithelialization was complete and the scab rejected. In general, collagenase shows an early maximum and then declines with postoperative time, with the sharpest decline occurring when epithelialization is complete.
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PMID:Collagenase in wound healing: effect of wound age and type. 146 86

The involvement of external Ca2+ in channel activity of nicotinic acetylcholine (nACh) receptors was investigated in the dissociated skeletal muscle fibers of adult mice. Single cells were prepared from flexor digitorum brevis muscles by treatment with collagenase and trypsin. Acetylcholine receptor-channel currents were recorded at endplates in the cell-attached mode, using the patch-clamp technique. Opening frequency, opening pattern and channel conductance were measured at different concentrations of ACh and succinylcholine (SuCh), using a patch pipette. Bath-applied ACh (30 microM) and SuCh (3-30 microM) decreased the ACh (1 microM)-induced channel conductance (SuCh was more potent than ACh). Large concentrations of both ACh (100-1000 microM) and SuCh (30-300 microM), which were applied via a patch pipette, also decreased channel conductance in a concentration-dependent manner. Increasing the concentration of calcium [Ca2+]0 in the patch pipette from 0 to 10 mM CaCl2 reduced ACh (1 microM)- and SuCh (1 microM)-activated single channel conductances. The increase in [Ca2+]0 prolonged the mean open times by 34% for ACh and by 22% for SuCh and decreased the channel opening frequency by 50% and 60%, respectively. These results demonstrate that ACh receptor-channel properties are dependent on [Ca2+]0 and that the ACh- and SuCh-induced decrease in channel currents may be also related to intracellular concentration of Ca2+. The effect of SuCh was greater than that produced by ACh.
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PMID:External Ca2+ dependence of acetylcholine- and succinylcholine-induced changes in channel conductance, open time and frequency at endplates of single muscle cells of adult mice. 166 97

Immature female Sprague-Dawley rats were primed with 20 IU eCG at 28 days of age and treated with 10 IU hCG 48 h later. Ovulation followed at 12-14 h. Ovaries were extracted at various times after hCG by use of Triton X-100 and 10 mM CaCl2 (Triton extract) followed by heating to 60 degrees C for 6 min with 0.1 M CaCl2 in 50 mM Tris/0.15 M NaCl, pH 7.5 (heat extract). These extracts were tested for their ability to inhibit tissue metalloproteinases by use of the small uterine metalloproteinase (UMP) of the rat. The ovary contains three plasma-derived inhibitors (alpha 1-macroglobulin [alpha 1 M], alpha 2-macroglobulin [alpha 2 M], and alpha 1 inhibitor3 [alpha 1I3]) and one tissue-derived inhibitor of the tissue inhibitor of metalloproteinases (TIMP) family. alpha 1 I3 was shown to inhibit UMP and rat collagenase. The concentration of the tissue inhibitor rose 5-fold from 0.6 micrograms UMP blocked per gram wet tissue in ovaries not primed with eCG to 3.2 micrograms UMP blocked at 8 h after hCG. Over this same time interval, the sum of alpha 1M + alpha 2M per gram of ovary rose 7-fold from 3.2 to 22.4 micrograms UMP inhibited and alpha 1I3 rose 2-fold from 4.4 to 10.7 micrograms UMP inhibited. The increases in the tissue inhibitor are interpreted to be due to increased synthesis by the tissue, whereas the changes in alpha-macroglobulins are postulated to be due to increased vascularity and increased permeability of the vessels.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:A tissue inhibitor of metalloproteinases and alpha-macroglobulins in the ovulating rat ovary: possible regulators of collagen matrix breakdown. 172 97

The studies included here identify factors affecting cartilage digestion by crude bacterial collagenase (cCGN) and describe a cartilage digestion medium that maximizes both tissue digestion rate and viable cell yield. The basal digestion medium contained 100 mM NaCl, 3 mM K2HPO4, 1 mM CaCl2, 1 mM MgSO4, 10 mM NaHCO3, 60 mM sorbitol, 5 mg/ml of dextrose, 1 mg/ml of albumin, and 2 mg/ml of cCGN in 25 mM HEPES at pH 7.2. Approximately 45% of articular cartilage tissue was digested in this basal medium in 6 h at 37 degrees C, yielding 6.8 x 10(6) viable cells per g tissue digested. The addition of 30 microM tosyllysylchloromethane (TLCM) increased the fraction of tissue digested in 6 h to 68% (p less than 0.05) and doubled viable cell yields to 13.6 x 10(6) per g tissue digested (p less than 0.05). Withholding Mg, decreasing NaCl to 70 mM, and adding 30 mM KCl increased fractional tissue digestion to 81% (p less than 0.01) and doubled viable cell yield yet again (to 29.9 x 10(6) viable cells per g tissue digested). Supplementation with TLCM increased the rate of cartilage digestion and the yield of viable cells regardless of cCGN source or lot. Additional trypsin (0.25%) inhibited tissue digestion and decreased cell yield; this effect was reversible with the addition of TLCM. The cartilage digestion medium developed in these studies (low Mg with added K and TLCM) was very effective in digesting articular, scapular, rib, and growth plate cartilage, as well as in yielding a large number of viable chondrocytes. These cells grew well in culture and maintained their chondrocytic characteristics, secreting predominantly type II collagen and large macromolecular forms of chondroitin sulfate-rich proteoglycans.
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PMID:Enzymatic isolation of chondrocytes from immature rabbit articular cartilage and maintenance of phenotypic expression in culture. 185 87

Oocytes of 40% of Xenopus laevis frogs respond to acetylcholine (ACh). Oocytes of the majority of responders exhibit the common two-component depolarizing muscarinic response (mean amplitude of the rapid component, 54 nA). Oocytes of approximately 10% of the responders ("variant" donors) exhibit a muscarinic response characterized by a very large transient, rapid current (mean amplitude 1242 nA, reversal potential -33 mV). Responses in oocytes of variant donors exhibit further qualitative differences: pronounced desensitization (absent in oocytes of common donors), characteristic prolonged latency (5.4 vs 0.9 s in oocytes of common donors) and marked inhibition of the response by activators of protein kinase C. Rapid responses in oocytes of variant donors are usually increased by treatment with collagenase, which, in common oocytes, often results in a complete loss of the response that correlates with the loss of muscarinic ligand binding. The number of muscarinic receptors was similar in oocytes of both types of donors (2.2 vs 3.0 fmol/oocyte). Also, the responses of oocytes of variant donors to microinjections of CaCl2 or inositol 1,4,5-trisphosphate were similar to those found in cells of common donors. These findings imply that altered receptor number, calcium stores and/or chloride channel density are not responsible for the variant responses. However, ACh caused an sixteen-fold greater efflux of 45Ca in oocytes of variant donors (35 vs 2.2% of total label in oocytes of common donors). Hence, the characteristics of the variant response may be related to a more efficient coupling between receptor stimulation and the mobilization of cellular calcium.
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PMID:Two types of intrinsic muscarinic responses in Xenopus oocytes. I. Differences in latencies and 45Ca efflux kinetics. 196 11

Investigation about the histamine receptors of gastric smooth muscle cells was done using isolated gastric smooth muscle cells of guinea pig. Isolated gastric smooth muscle cells were prepared from the stomach of guinea pig by percoll density gradient centrifugation after enzymatic digestion with collagenase and dyspase. Contraction of the cells was determined by image of phase-contrast microscopy and digitizer and expressed as the mean per cent of control in cell axis length. The concentration of intracellular Ca2+ ([Ca2+]i) was measured with the calcium-sensitive dye fura-2. Tested drugs were added to the incubation medium containing 2 mM CaCl2. Both contractile response and [Ca2+]i increased following histamine stimulation in a dose-dependent manner from 10(-7) to 10(-4) M, and those peaks were obtained in application of 10(-4) M pyridylethylamine, a H1 receptor agonist, made an increase in [Ca2+]i as much as 10(-4) M histamine. But 10(-4) M dymaprit, a H2 receptor agonist, made only small increase in [Ca2+]i, whose value was similar to that in application of only 2 mM CaCl2. 10(-4) M pyrilamine, a H1 receptor antagonist, inhibited the response to 10(-4) M histamine, and the value was similar to that in application of only 2 mM CaCl2. Cimetidine, a H2 receptor antagonist, did not inhibit the response to 10(-4) M histamine stimulation at all. From those results it is concluded that the action of histamine to the gastric smooth muscle cells of guinea pig may be mediated through H1 receptor, but not through H2 receptor.
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PMID:[Studies on the contractile mechanism of isolated gastric smooth muscle cells of guinea pig--investigation of the histamine receptor]. 198 90


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