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)

A method for isolating and culturing osteoclast-like cells from cancellous bone material collected from external iliac crest bone of patients is described. Aseptic techniques were used for comminution of the bone material, treatment with collagenase and separation of the bone cells from the bulk bone through a nylon filter. The bone cells were cultured on various surfaces for ten days. Cell motility, mobility and fusion was be observed along with tartrate-resistant acidic phosphatase activity in a majority of the cells soon after they had been cultured. These large cells attached to human cortical bone fragments, where they produced resorption lacunae in vitro. These morphologic and functional characteristics indicate that the cells we had isolated were, in fact, human osteoclasts. SEM studies of these cells on various biomaterials (titanium, hydroxyl apatite, tricalcium phosphate) revealed different morphologic characteristics varying with the substrate used and allowing conclusions as to substrate acceptance. Large areas of cell contact and cell proliferation suggest a favorable response to the materials applied.
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PMID:[Cell cultures of human osteoclasts for testing biomaterials]. 217 62

An improved sensitive assay for collagenase, which uses [3H]telopeptide-free collagen as a substrate, was used to measure changes in serum collagenase levels in 96 women and ten men (18-35 years old). Both latent and active forms of collagenase were detected in serum by molecular sieve chromatography; these forms had a relative molecular weight (Mr) of 65,000 and 45,000, respectively. Only latent collagenase was detected in crude serum after destroying inhibitors by treatment with 3 M potassium thiocyanate. Collagenase levels in males were lower than in nongravid females (34 +/- 5 versus 53 +/- 5 U/dL; mean +/- SEM; 1 unit = 1 microgram collagen digested per minute at 30C). During pregnancy there was no significant change in serum collagenase levels until the onset of spontaneous labor in full-term pregnancies (37-42 weeks), at which point there was a 66% increase over the nongravid level to a value of 88 +/- 5 U/dL. There was a further rise at one day postpartum, and high levels continued for at least four days. Women in premature labor (24-36 weeks) exhibited an eightfold increase in the level of serum collagenase to 405 +/- 110 U/dL; 16 of 17 patients in this group had collagenase levels above the 95th percentile for women at 16-40 weeks but not in labor. This evaluation of serum collagenase may provide a key for detecting premature labor. It is suggested that the increase in serum collagenase arises from the lower uterine segment and cervix.
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PMID:High levels of serum collagenase in premature labor--a potential biochemical marker. 243 51

In vitro perfused kidneys of ovalbumin-sensitized guinea-pigs consistently released relatively large quantities of histamine when challenged with the specific antigen (mean +/- SEM in twelve experiments was 37.7 +/- 6.0% of total kidney histamine, maximum 70.6%, compared with a basal release of 0.5 +/- 0.46% over a comparable period) but not with non-cross-reacting antigens. There was also no release from non-sensitized kidney. Rabbit antisera to guinea pig IgG1 and IgG2 immunoglobulins (but not normal rabbit serum) also consistently released histamine from perfused kidneys of sensitized guinea-pigs, but the release was smaller than with antigen, and could also be obtained from kidneys of non-sensitized guinea-pigs (maximum release 62.4% with the most potent antiserum). Guinea-pig kidney cell suspensions prepared by collagenase dispersion in vitro responded similarly, but the release with antigen was small (less than 10% net release, minus the spontaneous release 9.46% on average) as compared to anti-IgG1 (net release up to 38%) or anti-IgG2 (up to 44%). Rat kidney cells prepared by a similar procedure, and passively sensitized in vitro by incubation with rat immunoglobulin E (IgE) myeloma protein also responded to the addition of antiserum to rat IgE by releasing substantial amounts of histamine (up to 44% net release). In addition, heparin-containing cells (presumably mast cells or equivalent) in the enzyme-dispersed kidney cell preparations in both species were identified and counted by an adaptation of the Technicon H 6000 system used for counting blood basophils, and shown to represent 1 in 10,000 or less of the total cell population, which was not different from the count of similar cells in lung and heart tissues.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Renal histamine: release by immune stimuli. 243 15

A comparison was made between collagenase-dispersed guinea-pig atrial and ventricular tissues. Heparin containing cells were stained with alcian blue at pH 2.2, and counted by an automated technique (Technicon H6000). The cells were challenged with the specific antigen (ovalbumin), with antisera to guinea-pig IgG (non-subclass specific), IgG1 and IgG2, and the calcium ionophore A23187. Histamine release was measured by an automated spectrofluorometric technique, and leukotriene C4 was measured by radioimmunoassay. All of the following parameters were higher in the atrial than in ventricular cells (mean ratio and SEM of atrial: ventricular mast cell parameters in parenthesis): Histamine content/g wet tissues (3.32 +/- 0.71:1) (p less than 0.05), Absolute mast cell number as a proportion of total cell count (3.75 +/- 1.64:1), Histamine release induced by antigen (significant in one out of four experiments), anti-IgG (significant in three out of four experiments), anti-IgG1 (significant in two out of four experiments), and anti-IgG2 (higher but not statistically significant). Ionophore A23187 gave an inconsistent histamine release pattern: significantly higher release from atria in five treatments (different concentrations in different experiments), and higher ventricular release in three. Significantly more leukotriene C4 was released by antigen and the ionophore A23187 (mean of 3-5 treatments), but not with anti-IgG.
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PMID:Comparison of the response of mast cells in guinea-pig cardiac atria and ventricles. 243 72

For the improvement of the adult osteoblast culture, the osteoblasts of young adult rabbit endosteal from long bones were isolated by collagenase digesting separation. 0.1% of type-I collagen precoated culture flasks were used as substrate for isolated bone cell growth. Morphological examination of cultured cells under a phase-contrast microscope, SEM and TEM observations showed a structure similar to osteoblast in vivo. Histochemical examination of alkaline phosphatase demonstrated 97% purity of cultured osteoblasts. The presence of calcium deposit activity in cultured cells was demonstrated by Van Kossa stain. High activity of alkaline phosphatase and inorganic pyrophosphatase in cultured osteoblasts as determined by biochemical analysis. High calcium uptake in cultured osteoblasts was demonstrated by radioisotope labelled 45CaCl12. According to these methods, it was indicated that the cells isolated from young rabbit long bone endosteal were osteoblast-like and still maintained their biological function. Our system for culturing osteoblast-like cells is a successful attempt in growing bone tissue in vitro starting from isolated bone cells. Therefore, this modified method for bone cell culture on collagen precoated culture flasks could be used as the experimental model in studies concerning the osteoblasts in vitro.
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PMID:Functional evaluation of cultured rabbit osteoblast-like cells. 255 21

Pancreas obtained from 34 adult human cadaver organ donors was divided into proximal and distal segments, and the duct to each segment was cannulated. Collagenase was injected into the proximal duct of 7 glands and into the distal duct of 7 others; the duct of the opposite segment was perfused with collagenase. The pancreas was then dispersed by teasing, trituration, and passage through filters. Perfused proximal and distal segments released 1461 +/- 287 and 2728 +/- 797 islets/g (+/- SEM) versus 710 +/- 149 (P less than 0.05) and 1950 +/- 636 after injection. Twenty other pancreases were perfused with collagenase warmed rapidly to 39 degrees C (n = 4) or warmed slowly to 37 degrees C (n = 6) or 39 degrees C (n = 10): the yield was 1625 +/- 632, 1320 +/- 116, and 2009 +/- 277 islets/g respectively. Total yields from the latter were 76 X 10(3) large (greater than 100 microns) and 85 X 10(3) small (less than 100 microns) islets with recoveries of 61% and 42%, respectively, after Ficoll density gradient purification. Histology showed highly purified islets. Perifusion with glucose elicited a biphasic release of insulin with the mean response (microU/islet/min) rising to a first peak of 0.5 and constant second phase secretion of 0.25, followed by a return to baseline. Reduced response was observed for islets from pancreas stored greater than 6 hr and tissue obtained from multiple centers. Less insulin was produced by freshly isolated islets, islets less than 100 microns, and after Ficoll separation. Secretion was similar for islets derived from proximal or distal segments. Perfusion of collagenase via the ducts of human pancreas improves islet isolation and Ficoll gradient separation yields highly purified islets. Important factors influencing insulin secretion are the source of donor tissue, cold storage of pancreas, Ficoll purification, islet size, and tissue culture.
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PMID:Studies of the isolation and viability of human islets of Langerhans. 283 43

A method for estimating the concentration of spermatozoa in the rat cauda epididymidis is described. Treatment of a sperm suspension with 0.05% collagenase for 20-60 min or 0.025% trypsin for 1-2 min at 34-37 degrees C was found to result in consistently homogeneous sperm. Sperm concentration ranged from 152.5 to 230.0 X 10(7) spermatozoa/ml, with a mean of 187.7 (+/- 5.6 SEM) X 10(7) spermatozoa/ml.
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PMID:A method for estimating the concentration of spermatozoa in the rat cauda epididymidis. 283 33

Seven biopsy specimens from the cervix and 17 from the lower uterine segment were obtained in 24 women at term (37 to 42 weeks). Collagenase was extracted and assayed on telopeptide-free [3H]collagen; typical collagen cleavage products were found on sodium dodecyl sulfate-gel electrophoresis. There was no significant difference between collagenase levels in the cervix and in the lower uterine segment in women not in labor and with the cervix closed. Levels of active and latent collagenase in 11 such specimens were 0.14 +/- 0.03 and 0.64 +/- 0.90 U/gm wet weight, respectively (mean +/- SEM; 1 U = 1 micrograms collagen digested per minute at 30 degrees C). Thirteen women at term in active labor with cervical dilation of 4 to 8 cm exhibited a thirteenfold increase in mean collagenase activity in the lower uterine segment. Active and latent collagenase increased to 2.06 +/- 0.92 and 8.64 +/- 2.87 U/gm, respectively. This is the first direct evidence that interstitial collagenase increases markedly during cervical dilation in human parturition.
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PMID:Elevated tissue levels of collagenase during dilation of uterine cervix in human parturition. 284 86

Adrenocortical function was assessed in six normal and six chronic (greater than 12 weeks), DOCA-hypertensive Yucatan miniature swine; mean arterial pressures were 115.3 +/- 11.7 and 163.6 +/- 27.2 mm Hg, respectively (mean +/- SEM). Adrenocortical function was evaluated in vivo by measuring changes in plasma cortisol and aldosterone in response to exogenous ACTH (0.25 mg, iv), and in vitro by measuring the responses of collagenase-isolated adrenocortical cells to ACTH and angiotensin II. Corticoids were measured by specific radioimmunoassay. Basal plasma cortisol values of conscious DOCA-hypertensive swine were approximately 53% of the values of normotensive swine (P less than 0.05). However, ACTH induced a 419% increase in plasma cortisol values in DOCA-hypertensive swine compared to a 261% increase in the normotensive swine (P less than 0.05). These differences between the two groups were not altered by anesthesia. There were no significant differences in ACTH-induced changes in plasma aldosterone between the normotensive and DOCA-hypertensive swine. Experiments in vitro showed that the corticoid secretory responses of adrenocortical cells from DOCA-hypertensive animals were 6 times more sensitive to ACTH and 3.2 times more sensitive to angiotensin II than those of cells from normotensive swine. Thus, despite the possibility of adrenocortical insufficiency due to suppressed plasma renin activity and the negative feedback of DOCA on the hypothalamic-hypophyseal-adrenal axis, adrenocortical function of DOCA-hypertensive swine was hyperresponsive to trophic hormones. Results from this study suggest that the DOCA-hypertensive swine may be a valuable model in elucidating the relationship between hypertension and adrenocortical function and in investigating nonclassical control of the adrenal cortex, that is, control exerted during the hypertensive state that exists apart from or in addition to that exerted by ACTH and angiotensin II.
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PMID:Adrenocortical function in deoxycorticosterone acetate (DOCA)-hypertensive Yucatan miniature swine. 298 91

A novel technique has been developed for microstructural studies of bone. The spatial organizations of the mineral and collagen fibres in bone have been a matter of discussion for some time, with numerous diverse observations arising from various preparative techniques. In this latest investigation details of the mineral structure are clearly revealed in the SEM by treating a cut and polished surface of bone with collagenase to remove the major organic component. This new procedure has minimal effect on the mineral and hence reveals microstructural detail which is far closer to that in vivo than in previous investigations. This paper concentrates on two aspects of the studies, namely the detailed morphology of the mineral component and the arrangement of the collagen fibres in the osteons of compact bone. Firstly, the mineral component is revealed as comprising 'crystallites' (approximately 20 nm diam.) which aggregate to form larger contiguous 'spheroidal particles' (approximately 100 nm diam.), which in turn form 'granules' (approximately 500 nm diam.). Secondly, the regions from which collagen fibres have been removed are clearly revealed, showing that within an osteon, alternating lamellae have collagen fibres oriented approximately parallel to and circumferential to the Haversian canal respectively.
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PMID:Bone microstructure by collagenase etching. 298 60


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