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)

To determine intracellular pH gradients rabbit renal cortical tubular cells were prepared by collagenase separation, suspended in a Krebs-Ringer buffer solution, and gassed with 95% O2-5% CO2 in a special nuclear magnetic resonance (NMR) probe. Renal tubular cellular pH was determined simultaneously from the distribution of 14C-dimethadione (DMO) (pHDMO) or the chemical shift of inorganic phosphate (pHNMR). Experiments were performed at different external pH values (pHe) ranging between 6.52 and 7.20. pHNMR, a measure of cytoplasmic pH, changed by an amount equal to the change in pHe. pHDMO, however, a measure of cytoplasmic plus mitochondrial pH, changed less than pHe as the latter increased. pHDMO, higher than pHNMR at low pHe, became equal to pHNMR at higher pHe values. By use of assumed mitochondrial volumes of 30-40% mitochondrial pH was calculated from pHDMO and pHNMR. Mitochondrial pH remained relatively constant over the entire pHe range studied. Since cytoplasmic pH fell as pHe was lowered, the transmitochondrial pH gradient increased at low pHE values. These findings suggest that the transmitochondrial pH gradient may be important in regulating metabolism.
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PMID:Estimation of cellular pH gradients with 31P-NMR in intact rabbit renal tubular cells. 647 6

The isolated islets of Langerhans are the most available donors for transplantation. As the preservation of the isolated islets is difficult, we attempted to keep these tissues viable by use of an organ culture. Islets of Langerhans from adult Wistar rats were isolated by a collagenase technique and cultured in air-CO2 (95-5%) incubator at 37 degrees C. Insulin contents of the culture media which was changed every 3 days ranged from 1097 to 1434 microunits/ml during the 80 days' culture period. Transplantation of these islets into the portal vein of streptozotocin-induced diabetic rats resulted in a good recovery from the diabetic state. These studies indicate that cultured islets do preserve their original biological abilities.
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PMID:Culture of the islets of Langerhans for transplantation. 680 49

In this paper the theoretical basis of alloreactivity and its relevance to transplantation biology is discussed prior to a review of work showing that culture of adult mouse pancreatic islets for 7 days in 95% O2 and 5% CO2 facilitates successful grafting to nonimmunosuppressed allogeneic recipients. These allografts function by reversing both chemically induced and spontaneous diabetes. The fetal mouse pancreas is more immunogenic than adult islets, and even after a culture period of 10 days in 95% O2 and 5% CO2, BALB/c allografts are consistently rejected by nonimmunosuppressed recipient mice. The immunogenicity of fetal pancreas is thought to be due to the presence of contaminating lymphoreticular cells in the mesentery surrounding the fetal pancreas. Digestion of the fetal pancreas with collagenase allows the isolation of proislets that develop into functional islet tissue on transplantation. Fetal proislets are less immunogeneic than the whole fetal pancreas and may provide a source of tissue for clinical transplantation. Established islet allografts are relatively stable and are not rejected following nonspecific stimulation of the recipient's immune system or following passive transfer of either antibody or antibody and complement. After prolonged residence in the recipient a state of allograft tolerance develops and such grafts resist rejection by specific stimulation of the recipient. The administration of donor antigen in the form of uv-irradiated cells enforces this state of allograft tolerance.
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PMID:The reversal of diabetes by pancreatic islet transplantation. 681 62

In order to study the hormonal characteristics of human prolactin-secreting pituitary adenomas, in vitro monolayer and suspension cultures from human pituitary glands were established. Optimal conditions for cultures included enzymatic dispersion into viable single-cell suspensions with the use of 1% collagenase in phosphate-buffered saline solution. After pelleting the dispersed cells by centrifugation (800 rpm for 10 minutes), they were cultured in RPMI medium that contained 20% fetal calf serum and then incubated at 37degrees C in 5% CO2. Cells were subcultured weekly at a ratio of one plate to two. In an attempt to establish whether bromocriptine has a direct inhibitory effect on pituitary secretion of prolactin (PRL), variable doses of bromocriptine were added to duplicate plates. The addition of bromocriptine to the culture medium induced suppression of PRL within 7 days. In conclusion, this study demonstrated that either monolayer of suspension cultures of human PRL secreting adenomas can be established, and that bromocriptine in doses of 1 ng/plate or more has a direct inhibitory effect on the secretion of PRL.
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PMID:Monolayer and suspension culture of human prolactin-secreting pituitary adenoma. 743 30

The factors determining the outcome of human fetal islet transplantation in patients with insulin-dependent diabetes mellitus (IDDM) remain unclarified. In this study we analysed the ratio between immunoregulatory lymphocyte subpopulations in order to search for a possible marker of the immune destruction of transplanted islets. Human fetal islets were isolated by collagenase digestion, cultured for 14 days at 37 degrees C, 5% CO2, and implanted under fascia of m. rectus abdominis in 7 IDDM patients (5 pancreata per patient). After transplantation we evaluated simultaneously the level of metabolic control through HbA1c values determined by chromatography, the capacity of insulin secretion through the C-peptide levels (determined by radioimmunoassay) before and 6 minutes after 1 mg glucagon i.v. stimulation, and the ratio between CD4+ and CD8+ lymphocytes determined by immunofluorescence using monoclonal antibodies. We found that metabolic control after transplantation was improved together with the decrease of the insulin daily dose, and the improvement was simultaneous to the increase of both basal and glucagon-stimulated C-peptide levels. Four months after transplantation we detected a remarkable decrease in the secretion capacity, accompanied by the necessity for an increase in daily insulin dose to maintain optimal metabolic control. However, the loss of islet function was preceded by the increase in CD4+/CD8+ ratio, thus reflecting the presumable accumulation of CD4+ inducer T-lymphocytes. When the islet secretion capacity was destroyed, we found a decrease in CD4+/CD8+ ratio, reflecting the recruitment of CD8+ effector cells.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:[Human fetal pancreatic islet transplantation in insulin-dependent diabetics: possibilities of early detection of transplant destruction]. 759 Apr 18

On the basis of earlier reports of reduced growth rate and fat accumulation in animals fed trans 18:1, a study was conducted to determine whether the effects of trans 18:1 on lipolysis and glucose utilization by adipocytes differed from effects of the cis isomer. Two experiments compared three fatty acid isomers (oleic, elaidic and vaccenic acids) at several concentrations and at several fatty acid to albumin ratios in cell media. Adipocytes were isolated from adipose tissue of rats by collagenase digestion and incubated for 2 h in media containing added fatty acids. Compared with oleic acid, both trans isomers reduced (P < 0.01) the amount of glucose converted to cell lipid in both experiments. Glucose oxidation to carbon dioxide also was lower for both trans fatty acids in Experiments 1 (P < 0.05) and 2 (P < 0.06). Lipolytic rates were increased (P < 0.01) in both experiments by replacing oleic acid with either of the trans isomers. Trans isomers of octadecenoic acid had catabolic effects on adipocyte metabolism that occurred regardless of the position of the double bond, the fatty acid concentration in media or the fatty acid to albumin ratio. These catabolic effects explain previous observations of reduced growth rate and fat accumulation when oleic acid in animal diets is replaced with a trans isomer.
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PMID:Replacing cis octadecenoic acid with trans isomers in media containing rat adipocytes stimulates lipolysis and inhibits glucose utilization. 766 58

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


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