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)

Stromal-vascular (S-V) cells isolated from adipose tissue of newborn pigs (NBPC) and mature pigs (MPC) by collagenase digestion were used to evaluate differences in preadipocyte culture and development. Cells were seeded at a density of 3 x 10(4) cells/cm2 on six-well (35-mm) tissue culture plates in 3 mL of DMEM/HAM's F12 medium plus 10% fetal calf serum and cultured at 37 degrees C under a humidified atmosphere of 95% air:5% CO2 for 24 h. Cells were then washed thoroughly in DMEM/HAM's F12 medium without fetal calf serum and maintained in serum free (SF) medium or SF medium supplemented with 2.5% newborn pig serum (NBPS) or mature pig serum (MPS) for 12 d. After 1 d, more NBPC adhered to the culture plates, as indicated by DNA values. After 12 d, protein per culture well was not significantly different, but DNA concentration per well remained higher (P < .05) in cultures of NBPC than in the MPC cultured in the same medium, indicating fewer MPC. Protein:DNA ratios were higher (P < .05) in cultures of MPC regardless of the medium, reflecting larger cell size. More cells containing fat deposits were seen with NBPC in all conditions in comparison with MPC, and more fat was deposited in NBPC in SF than in SF plus NBPS or MPS. The NBPC had higher (P < .05) sn-glycerol-3-phosphate dehydrogenase (GPDH; EC 1.1.1.8) per protein than MPC regardless of the medium. For both cell types, GPDH activity in either serum was less than activity of cells grown in SF.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Effect of age on the differentiation of porcine adipose stromal-vascular cells in culture. 773 Jan 75

Three isolates of a previously undescribed Dermatophilus sp. obtained from chelonids (two strains obtained from turtles and one strain obtained from a tortoise) were compared with 30 Dermatophilus congolensis isolates obtained from Australian mammals. The microscopic appearance, the colony morphology, and most biochemical test results for the chelonid isolates were characteristic of the genus Dermatophilus. Our isolates differed from the mammalian D. congolensis isolates in a number of cultural characteristics, including faster growth at 27 degrees C than at 37 degrees C, formation of two hemolysis zones around colonies on blood agar at 37 degrees C in the presence of 10% CO2, poor motility, and production of a distinctive odor. The DNA restriction enzyme digestion and protein electrophoresis patterns of our strains were distinct. The electrophoretic mobilities of 11 enzymes differed from the mobilities observed with D. congolensis strains. A monoclonal antibody to a surface antigen of an ovine isolate did not react with zoospores or filaments of the chelonid isolates. Biochemical differences between our isolates and D. congolensis included the ability of the chelonid isolates to reduce nitrate to nitrate and the fact that the chelonid isolates exhibit collagenase activity in vitro. We propose that the chelonid isolates should be placed in a new species, Dermatophilus chelonae. Strain W16, which was isolated from a nose scab on a snapping turtle, is the type strain; a culture of this strain has been deposited in the American Type Culture Collection as strain ATCC 51576.
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PMID:Dermatophilus chelonae sp. nov., isolated from chelonids in Australia. 785 7

The isolated bovine adrenocortical cells are prepared aseptically by the use of collagenase and deoxyribonuclease. The isolated cells are suspended in Ham F-10 medium containing 5% fetal calf serum, 10% newborn calf serum, 2.5% horse serum and antibiotics. The seeded cells are cultured at 37 degrees C in a humidified atmosphere of 5% CO2 in air. Steroidogenic activity for ACTH reached the maximum in the 2- to 3-day primary cultured cells; the maximum response to ACTH in these cells is more intense than that in freshly isolated bovine adrenocortical cells. The primary cultured cells have prostaglandin, muscarinic, ATP and beta-adrenergic receptors that are linked to steroidogenesis in addition to ACTH and aldosterone receptors. Thus primary cultured bovine adrenocortical cells are a useful tool to study these receptors and the intracellular events that are associated with the receptors. We also demonstrated that the fura 2 loaded primary cultured monolayer cells on glass cover slips provide us much more information than suspended cells in the study of intracellular Ca2+ mobilization in adrenocortical cells.
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PMID:[Primary culture of bovine adrenocortical cells]. 811 88

In our clinical use of lasers, mainly CO2 laser for oral surgery, we found that the laser had many advantages over an electrome and the laser improved the local control rate for malignant tumors. Low-power laser has been used to treat hypersensitive dentin, to relieve pain caused by neurotic disease around mouth, and to promote the healing of those diseases. The results obtained from the clinical applications showed that irradiation of the hypersensitive dentin with low-power laser was significantly effective in desensitization. An in vitro study showed no effects of diode or He-Ne laser irradiation on the growth of cells, but showed changes in the initial cell adhesion rate. He-Ne laser irradiation to the wound in the skin of hamsters caused to change the activities of the types I and III collagenase. This fact suggest that laser irradiation acted to promote the healing of wound.
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PMID:Clinical applications and basic studies of laser in dentistry and oral surgery. 812 80

The urinary bladder of Bufo marinus excretes H+ and this excretion is increased by metabolic acidosis (MA), insulin (IN), prostaglandin E2 (PGE2), increases CO2, and aldosterone. The purpose of this experiment was to determine whether MA, IN, PGE2, CO2, and aldosterone stimulate inositol phosphate's (IP) formation in isolated cells of toad urinary bladder. Cells were prepared by treating bladder sacs with collagenase. Cells were obtained from 10 toads in MA and 10 normal toads, suspended in 2 ml of Ringer's solution containing LiCl (10 mM), myo-inositol (5 mM), and [3H]myo-inositol (10 microCi), and then incubated for 2 hr at 25 degrees C. Cells were homogenized and the IP fractions quantitated by column chromatography and liquid scintillation counting. The results were expressed as dpm (mu MPO4)-1 (hr)-1. The IP in MA cells was 44,202 +/- 4,646 and in normal toad cells it was 31,637 +/- 3,613 (P < 0.05). In a separate experiment, cells from 10 paired hemibladders were isolated from normal toads. The cells were treated exactly as above except there were no LiCl in the bath. LiCl was added to all baths after 2 hr and the experimental cells were challenged with IN, PGE2, increases CO2, and aldosterone for 20 min. The IP were quantitated as above. IN treatment stimulated inositol bisphosphate and inositol triphosphate (P < 0.01). PGE2 and increases CO2 also stimulated inositol triphosphate (P < 0.05). Aldosterone did not alter formation of any of the IP fractions. We conclude that MA, IN, PGE2, and increases CO2 stimulate IP formation in cells of toad urinary bladder and inositol triphosphate may be an important second messenger in mediating the response of MA, IN, PGE2, and increases CO2.
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PMID:Stimulation of phosphoinositides by agents that stimulate proton secretion in toad urinary bladder. 841 76

The release of secretin was studied in secretin cell-enriched preparations isolated from canine duodenal mucosa. The crude enterocytes were isolated by treating the duodenal mucosa sequentially with collagenase and ethylenediaminetetraacetic acid. Secretin cell-enriched fraction was prepared by centrifugation of the crude enterocytes in a counterflow elutriation rotor to obtain a final preparation containing 3.2 +/- 0.3 pmol/10(6) cell of immunoreactive secretin, which was 13-fold greater than the crude cell preparation (N = 5). The cells were incubated in Hanks' balanced salt solution for 20 min at 37 degrees C under 95% O2/5% CO2 before adding various agents and further incubated for various periods of time. The amounts of secretin released into the medium and retained by the cells were then determined by a specific radioimmunoassay. The release of immunoreactive secretin was increased dose-dependently over the control by dibutyryl cyclic-3',5'-adenosine monophosphate, forskolin, 4 beta-12-O-tetradecanoylphorbol-13-acetate, the synthetic serine protease inhibitor, camostat, and the calcium ionophore, A23187. The effects of forskolin, the phorbol ester, and A23187 were time-dependent and not observed at 4 degrees C. The release of immunoreactive secretin was also stimulated by KCl in high concentration and by sodium oleate. The effect of A23187 was abolished in a Ca(2+)-free medium, while those of dibutyryl cyclic-3',5'-adenosine monophosphate and forskolin were potentiated by 3-isobutyl-1-methylxanthine, which did not have a significant effect when added alone. These results indicate that the release of secretin is regulated by both Ca(2+)- and cyclic-3',5'-adenosine monophosphate-dependent mechanisms.2+ release.
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PMID:Characterization of secretin release in secretin cell-enriched preparation isolated from canine duodenal mucosa. 842 47

The hypothesis that general anesthetics protect energy reserves by decreasing energy demand is widely accepted but poorly substantiated. Isoflurane at clinical doses preserved adenosine triphosphate (ATP) levels in anoxic isolated hepatocytes. Specific inhibitors were used to block mitochondrial and/or glycolytic ATP formation to ascertain whether pathways of energy supply or demand, or both, were involved in ATP preservation by isoflurane. Hepatocytes were isolated from fed adult male rats after perfusing livers with Krebs buffer containing collagenase. Cells were incubated in Krebs buffer for 0-30 min at 25 degrees C under N2/CO2 (95%/5%) +/- isoflurane 0.63 mM in liquid phase. Oligomycin, iodoacetate, or fasting were used to block mitochondrial and glycolytic ATP formation. Under anoxia alone, ATP levels declined more slowly in the presence than in the absence of isoflurane, confirming the ATP-protective effect of isoflurane reported previously. With oligomycin plus iodoacetate blocking all ATP formation, ATP decline (representing pure ATP consumption) was not slowed by isoflurane. Isoflurane's protective effect recurred when glycolytic ATP supply was restored by incubating with oligomycin only. The protective effect was accompanied by increased lactate accumulation, and both effects-ATP preservation and lactate formation-were similarly dependent on isoflurane concentration. We conclude that the protective effect of isoflurane on energy status in anoxic isolated hepatocytes was not associated with reduced ATP demand but with enhanced ATP supply via stimulation of glycolysis.
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PMID:Isoflurane preserves adenosine triphosphate levels in anoxic isolated rat hepatocytes by stimulating glycolytic adenosine triphosphate formation. 863 2

This study employed immunocytochemistry and toluidine blue counterstaining to compare different procedures utilized in primary myoblast cultures, from which an optimal culture model for normal myoblasts could be derived. The growth characteristics of normal and dystrophic myoblasts were also investigated by means of this model. Results indicate that the requirements for an ideal myoblast culture should include a combined enzyme of 0.25% trypsin and 0.2% collagenase (type IV) (1:1), a preplating time of approximately 15-20 minutes, and a seeding density of 1 x 10(5) cells/ml. Furthermore, the mouse samples should be newborn mice. A better proliferative capacity of myoblasts was noted in an incubator with 10% CO2 coupled with Dulbecco's MEM plus 15% fetal calf serum. With regard to the growth characteristics of normal and dystrophic myoblasts, the doubling time of normal myoblasts was shorter than that of dystrophic myoblasts. In terms of the fusion percentage of myoblasts, dystrophic myoblasts tended to fuse earlier than normal ones, especially after 5 days in culture. The findings of this study are valuable in understanding the myogenesis of myoblasts under different culture conditions. The establishment of requirements for good growth of myoblast cultures will facilitate myoblast transfer therapy. Finally, the growth characteristics of normal and dystrophic myoblasts as well as variances in the proliferation and differentiation of these two types of cells are clarified.
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PMID:Growth characteristics of normal and dystrophic myoblasts in primary myoblast cultures. 893 42

We have shown that cutaneous burn injury impairs cardiac contractile performance; however, the mechanisms remain unclear. In this study, New Zealand White rabbits were anesthetized with isoflurane, given a full-thickness scald burn over 30% of total body surface area, and resuscitated with lactated Ringer solution (4 ml.kg-1.%burn-1 for 24 h); rabbits handled in an identical fashion were given a sham burn. Serum obtained from burned and control (sham-burned) rabbits was aliquoted and frozen at -70 degrees C until assay. Polymorphonuclear neutrophils (PMN) were isolated 24 h postburn from both sham and burned rabbits to yield preparations with > 95% PMN with > 95% viability. Cardiac myocytes were isolated by retrograde perfusion of hearts with Ca(2+)-free collagenase-Tyrode buffer, suspended in Krebs-Henseleit buffer containing 10% fetal bovine serum and 1.8 mM Ca2+, and incubated (1 x 10(5) cells/well) in a CO2 incubator under several experimental conditions, including buffer alone, buffer plus 10% burn serum, buffer plus 10% sham serum, or buffer plus either burn or sham PMN (25 x 10(5) cells/well). Myocyte viability (%) and creatine kinase (CK; units.ml-1.10(5) cells-1) were unchanged after incubation with sham plasma or sham PMN. Incubation of sham myocytes with burn plasma caused viability to fall (from 79 +/- 3 to 54 +/- 4%, P < 0.002), whereas CK rose (from 1,639 +/- 115 to 2,803 +/- 132 units.ml-1.10(5) cells-1, P < 0.01). Similarly, incubation of sham myocytes with burn PMN reduced viability (from 83 +/- 2 to 50 +/- 3%, P < 0.01), whereas CK remained unchanged (1,880 +/- 168 units.ml-1.10(5) cells-1). Our data indicate that circulating myocardial depressant factors after burn injury contribute to cardiac myocyte injury.
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PMID:Cellular basis for burn-mediated cardiac dysfunction in adult rabbits. 899 23

Paraquat (PQ) induces lung, liver and kidney damage. Since PQ mainly is eliminated by the kidney, the kidney damage is of particular importance to the outcome of PQ poisoning. The exact toxic mechanism of PQ is still unclear but it is assumed to involve redox cycling and formation of reactive oxygen species. In this study, further investigations on the toxic mechanism and metabolic effects of PQ were performed using isolated renal proximal tubules from rabbits. Proximal tubules were isolated using a combined iron perfusion and collagenase method. Suspended tubules were incubated for varying periods and concentrations of PQ at 25 or 37 degrees C in Krebs-Ringer phosphate buffer or HCO3-/CO2 buffer. The cytotoxic effect of PQ was evaluated by (1) markers of oxidative stress: status of glutathione (GSH/GSSG) and formation of malondialdehyde (MDA); and (2) markers of tubular metabolism: oxygen consumption (QO2), transport of 14C-p-aminohippuric acid (PAH) and 14C-tetraethylammonium (TEA). Using 0.5 and 5 mM PQ, the GSH/GSSG ratio decreased whereas formation of MDA increased indicating oxidative stress. PQ reduced the accumulation of PAH and TEA, the basal QO2 and the ouabain sensitive QO2 indicating inhibition of the Na/K-ATPase. Nystatin-stimulated QO2 was reduced by PQ, excluding inhibition of Na+ entry as a possible cytotoxic mechanism and suggesting mitochondrial injury. This was confirmed by measuring FCCP-uncoupled QO2. Thus high concentrations of PQ appear to disrupt mitochondrial electron chain transfer resulting in reduction of metabolic functions.
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PMID:The cytotoxic effect of paraquat to isolated renal proximal tubular segments from rabbits. 927 8


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