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

---

Query: EC:3.2.1.36 (hyaluronidase)
4,606 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The effects of all-trans retinoic acid on glycosaminoglycan (GAG) accumulation were determined in cultured primary human skin fibroblasts. Confluent cultures treated with retinoic acid accumulated less [3H]GAG than those without the compound, an effect with an apparent threshold of 10 nM which was dose dependent in the concentration range tested (0-10 microM). At 10 microM, the inhibition was 54%. Greater than 80% of the labeled macromolecular material was streptomyces hyaluronidase digestible in cultures labeled with [3H]acetate. The incorporation of H2[35S]O4 into chondroitin sulfate and dermatan sulfate was unaffected, as was total protein synthesis. Retinol also inhibited accumulation of [3H]GAG, but was far less potent. T3 and dexamethasone can inhibit [3H]hyaluronate synthesis. When retinoic acid was added to cultures treated with either of these hormones at concentrations that maximally inhibit [3H] GAG accumulation, there was a further decrease in the rate of macromolecular accumulation. The retinoic acid effect evolved over 24-48 h after addition to the culture medium. A pulse-chase study failed to demonstrate any effect on [3H]GAG degradation.
...
PMID:Retinoic acid inhibition of hyaluronate synthesis in cultured human skin fibroblasts. 230 23

The side-effects of "artificial ascites" induced with Dextran 60 (Makrodex 6%) as a mean of preventing adhesions were investigated in 47 patients (treatment group: 32 patients; control group: 15 patients) in whom microsurgery had been performed for infertility with adhesiolysis. On the day of surgery and the following four days 300 to 500 ml of Makrodex was instilled via an intraperitoneal catheter (7.5 ml/kg body weight on day of surgery; 5 ml/kg body weight on days 2 to 5). In addition, the patients received, on the day of surgery, single doses of 450 IU of hyaluronidase (Kinetin), 500,000 KIU of aprotinine (Trasylol) and 1 g of hydrocortisone acetate instilled intraperitoneally. In the group treated with Dextran, there was a significantly higher number of patients who felt unwell and had abdominal complaints and dyspnea. In six cases in the Dextran group a vulval edema was seen, and in 2 cases a thigh edema. A significant weight increase and elevation of central venous pressure occurred for the duration of the "artificial ascites" in this group. There were a few cases of bradycardia with frequencies of under 50 beats per minute. On the fifth p.o. day 75% of the patients in the Dextran group had a pleural effusion. Such changes were not observed in the control group. In view of these side-effects and the fact that it is still not proven that Dextran effectively prevents adhesions we no longer carry out this form of adhesion prophylaxis.
...
PMID:[Complications and side effects of artificial ascites for adhesion prevention]. 241 47

The association of hyaluronate with the surface of chondrocytes was examined by several approaches using primary cultures of chondrocytes derived from the Swarm rat chondrosarcoma. In culture, chondrosarcoma chondrocytes produced large pericellular coats, which can be visualized by particle exclusion, and which can be removed by Streptomyces hyaluronidase. Exposure of chondrocytes, which had been metabolically labelled with 3H-acetate, to exogenous hyaluronate or to Streptomyces hyaluronidase resulted in the release of 36-38% of the endogenous, labelled chondroitin sulfate from the cell layer into the incubation solution. These results imply that at least 37% of the cell layer chondroitin sulfate proteoglycan is retained there by an interaction with hyaluronate. Thus membranes were prepared from cultured chondrocytes and examined for sites which bind 3H-hyaluronate. Binding was observed and found to be saturable, specific for hyaluronate, of high affinity (Kd = approximately 10(-10) M), and destroyed by treating the membranes with trypsin. The 3H-hyaluronate-binding activity was inhibited competitively by hyaluronate decasaccharides but not by hexasaccharides or octasaccharides, indicating that the binding sites recognize a sequence of hyaluronate composed of five disaccharide repeats. The binding activity was partially purified from a detergent extract of chondrocyte membranes by ion exchange chromatography on DEAE-cellulose, followed by affinity chromatography on wheat germ agglutinin-agarose. Analysis of the partially purified binding activity by SDS-PAGE revealed five protein bands of 48,000-66,000 daltons in silver-stained gels. SDS-PAGE followed by Western blotting and exposure to monoclonal antibodies which recognize epitopes present in link protein and in the hyaluronate-binding region of cartilage proteoglycan revealed no immunoreactive protein bands in the partially purified material. We conclude that one mechanism by which hyaluronate associates with the chondrocyte surface may be via interaction with a membrane-bound hyaluronate-binding protein which is distinct from link protein and proteoglycan.
...
PMID:Membrane-associated hyaluronate-binding activity of chondrosarcoma chondrocytes. 247 51

Primary cultures of rabbit articular chondrocytes have been maintained for 10 days and labeled with [35S]sulfate, [3H]leucine, and [35S]cysteine in pulse-chase protocols to study the structure and hyaluronate binding properties of newly synthesized proteoglycan monomers. Radiolabeled monomers were purified from medium and cell-layer fractions by dissociative CsCl gradient centrifugation with bovine carrier monomer, and analyzed for hyaluronate binding affinity on Sepharose CL-2B in 0.5 M Na acetate, 0.1% Triton X-100, pH 6.8. Detergent was necessary to prevent self-association of newly synthesized monomers during chromatography. Monomers secreted during a 30-min pulse labeling with [35S]sulfate had a low affinity relative to carrier. Those molecules released into the medium during the first 12 h of chase (about 40% of the total) remained in the low affinity form whereas those retained by the cell layer rapidly acquired high affinity. In cultures where more than 90% of the preformed cell-layer proteoglycan was removed by hyaluronidase digestion before radiolabeling the newly synthesized low affinity monomers also rapidly acquired high affinity if retained in the cell layer. Cultures labeled with amino acid precursors were used to establish the purity of monomer preparations and to isolate core proteins for study. Leucine- or cysteine-labeled core proteins derived from either low or high affinity monomer preparations migrated as a single major species on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with electrophoretic mobility very similar to that of core protein derived from extracted proteoglycan monomer. Purified low affinity monomers were converted to the high affinity form by treatment at pH 8.6; however, this change was prevented by guanidinium-HCl at concentrations above 0.8 M. Conversion to high affinity was also achieved by incubation of monomers in aggregate with hyaluronic acid (HA) at pH 6.8 followed by dissociative reisolation of monomer. At both pH 6.8 and 8.6 the conversion process was slow, requiring up to 48 h for the maximum increase in affinity. It is suggested that the slow increase in HA binding affinity seen during extracellular processing of proteoglycans in cartilage and chondrocyte cultures is the result of an irreversible structural change in the HA binding domain following the binding of monomer to hyaluronate. The available evidence suggests that this change involves the formation or rearrangement of disulfide bonds.
...
PMID:Studies on the hyaluronate binding properties of newly synthesized proteoglycans purified from articular chondrocyte cultures. 249 59

Bilateral nephroblastomatosis was diagnosed in a 15-month-old white female. Prior to surgery, multiple peripheral blood smears (Wrights' stain) revealed an azurophilic staining extracellular material. When serum was added to a three percent acetic acid solution, a floccular, fibrous precipitate formed at the meniscus of the tube. Serum protein electrophoresis on cellulose acetate support media resulted in a distorted pattern which corrected to a normal pattern upon treatment with hyaluronidase. These peripheral blood abnormalities disappeared following a left nephrectomy. Quantitative chemical analysis of diseased renal tissue yielded 81 micrograms of readily extracted glycosaminoglycan (GAG) per gram of tissue. The importance of abnormal glycosaminoglycan production in patients with malignant disease is discussed both in terms of clinical importance and possible roles of cell exudates.
...
PMID:Glycosaminoglycan in the blood and renal tissue of a patient with nephroblastomatosis. 298 11

In neutrophils preincubated with non-inflamed synovial fluid, opsonized zymosan-induced O2- production was increased by 232.7 +/- 19.1% and degranulation induced by zymosan or phorbol myristate acetate was increased by 152.8 +/- 21.8% and 217.4 +/- 26.3 respectively. Unstimulated neutrophils were not directly activated by the fluid, nor did it affect their chemotactic response. The activity was not abolished by heat (56 degrees C, 30 min, or 100 degrees C, 3 min) or by treatment with trypsin or hyaluronidase. The activity was not present in the serum albumin-bound lipid extract of the synovial fluid. Thus, this activity may represent a heat- and trypsin-resistant factor which is neither a complement component nor hyaluronic acid. It is proposed that this synovial fluid factor plays a role in increasing the neutrophil killing potential during the inflammatory process.
...
PMID:The effect of non-inflamed synovial fluid on neutrophil function in vitro. 299 26

Methods for the analysis of urinary GAGs that can be used for or are applicable to routine assays are described. The most popular method for isolation of GAGs from a urine sample is CPC precipitation, in spite of the fact that it is time-consuming. To identify the different types of GAGs excreted, separation by one-dimensional cellulose acetate electrophoresis followed by staining with alcian blue or toluidine blue may suffice for routine purposes. Solvents such as barium acetate, calcium acetate, barbital buffer and pyridine-formic acid are used for the separation. However, the separation of the seven types of GAGs by conventional one-dimensional electrophoresis is difficult, and a discontinuous electrophoretic method with barium acetate buffer and barium acetate buffer containing ethanol has proved effective for the separation. HPLC separation methods are used for assaying the profiles of enzymatic digestion products of GAGs. Advanced HPLC methods for separating intact GAGs of different types are currently unavailable. Unsaturated disaccharides produced with heparitinase and/or heparinase from heparan sulphate and oligosaccharides produced by hyaluronidase digestion of hyaluronic acid can be separated by HPLC. For chondroitin sulphate isomers, unsaturated disaccharides produced by digestion of the samples with chondroitinase ABC or chondroitinase AC are separated by HPLC and determined by their UV absorbance or by fluorescence labelling. Highly sensitive quantitation of chondroitin sulphate isomers is possible by these methods, which are also efficient for the investigation of the constituents of GAG polymers. Some of these methods have been applied to urine samples from patients with, e.g., mucopolysaccharidoses.
...
PMID:Methods for analysis of urinary glycosaminoglycans. 306 22

The purpose of this study was to examine the nature of the linkage between cell-surface hyaluronate and the plasma membrane. To accomplish this, rat fibrosarcoma cells were cultured in the presence of [3H]-acetate to isotopically label the hyaluronate, and then fixed with glutaraldehyde, which cross-links proteins but does not react directly with hyaluronate. The glutaraldehyde fixation stabilized the cells so that they could be manipulated in ways which would otherwise destroy cells. The fixed cells were then subjected to various treatments, and the amount of hyaluronate remaining on the cell surface was assayed via exhaustive digestion with Streptomyces hyaluronidase. Using this technique, we found that 1) cell-surface hyaluronate was quite stable for extended periods of time even in the presence of a large excess of non-labeled hyaluronate; 2) 4 M guanidine HCl and detergents did not extract a significant portion of cell-surface hyaluronate; 3) solutions of varying ionic strength (0-1 M NaCl) had no effect on the retention of hyaluronate; 4) the cell coat was stable in the range of pH 4-11, but outside this range a significant amount of hyaluronate was released; and 5) treatment with proteases released cell-surface hyaluronate. These results are consistent with the possibility that hyaluronate is covalently linked to a protein associated with the plasma membrane. Further support for this model came from experiments with the detergent Triton X-114, which can be used to separate soluble proteins from hydrophobic proteins. When nonfixed rat fibrosarcoma cells were extracted with this detergent and then partitioned by centrifugation, approximately 30 times as much hyaluronate was present in the detergent fraction which contained the hydrophobic proteins, as compared to the extracts pretreated with trypsin prior to phase separation. Again, these results suggest that cell-surface hyaluronate is directly linked to a hydrophobic core protein intercalated in the plasma membrane.
...
PMID:Hyaluronate appears to be covalently linked to the cell surface. 312 2

Fibroblast cultures established from explants of mature scar and skin tissue were analyzed with regard to extracellular glycosaminoglycan (GAG) composition and response to interleukin-1 (IL-1). Following a serum-free 48 hour label with [3H]glucosamine, pericellular and medium GAGs were isolated by precipitation with cetylpyridinium chloride (CPC) and analyzed by cellulose acetate electrophoresis. In addition, susceptibility of the precipitates to Streptomyces hyaluronidase, chondroitinase ABC and heparitinase was determined. Labeled conditioned medium from the scar-derived cells contained both dermatan sulfate (DS) and hyaluronate (HA), as compared to medium from the control (skin-derived) cells which contained predominantly DS. IL-1 induced the appearance of chondroitin 4-sulfate (C4-S) in the medium of the scar cells with no concurrent effect on either DS or HA, and increased the amount of HA in the medium fraction of normal skin cells. The pericellular fraction of the scar-derived cells contained chondroitin 6-sulfate (C6-S) and DS; addition of IL-1 resulted in a shift from DS to heparan sulfate (HS), and the emergence of a pericellular GAG profile similar to that of normal dermal fibroblasts.
...
PMID:Interleukin-1-induced changes in extracellular glycosaminoglycan composition of cutaneous scar-derived fibroblasts in culture. 313 46

Gingival exudates from sites of acute ulcerative gingivitis (AUG) and chronic gingivitis (CG) in adults were investigated by cellulose-acetate electrophoresis for the hyaluronic acid (HA) content and assayed for the levels of HA-degrading enzymes. HA was the only glycosaminoglycan (GAG) in gingival exudate from CG sites. HA was not detected at untreated AUG sites but was evident, at increasing levels, after two and seven days of effective antibacterial treatment. In AUG exudates, the total HA-degrading enzyme activity, of bacterial origin, decreased to approx. 30 and 10 per cent of the high initial levels after two and seven days of treatment respectively, to that level found at sites of CG. The specific activity of HA-degrading enzyme of lysosomal origin was low initially and increased with treatment to a level comparable to CG. The notable absence of HA from gingival exudate from sites of untreated AUG thus appears to result from the increased levels of bacterial hyaluronidase. Electrophoresis of gingival exudate may be an indirect method of monitoring the rate of response of AUG to different antibacterial treatments.
...
PMID:Hyaluronic acid and hyaluronidase activity in gingival exudate from sites of acute ulcerative gingivitis in man. 332 9


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>

---