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

A new type of hyaluronidase was isolated from squid cranial cartilage. The enzyme seems to be localised extracellularly, since it is extracted from the tissue by 0.5 M sodium acetate, pH 7.0, in the presence of proteinase inhibitors. Degradation studies suggest that the enzyme belongs to the family of endoglycosidases generating oligosaccharides of rather large size. The best activity of the enzyme was observed at pH 7.0 and 37 degrees C and the optimum buffer for digestion was 0.15 M Tris acetate. It is inactive in sodium phosphate, morpholine acetate and HEPES buffers. The enzyme degrades aggrecan, hyaluronan, chondroitin sulphate and oversulphated chondroitin sulphate.
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PMID:The presence of a novel extracellular hyaluronidase in squid cranial cartilage. 1538 35

Neoplastic mast cells of mice (including long-established and newly derived lines) were grown in large-volume suspension cultures to provide enough cells for preparation of microsomal fractions. Microsomal preparations from P815Y and P815S cells synthesized (14)C-labelled glycosaminoglycan when incubated with UDP-[(14)C]glucuronic acid and UDP-N-acetylgalactosamine. No significant amount of (14)C-labelled glycosaminoglycan was formed when UDP-N-acetylglucosamine was substituted for the UDP-N-acetylgalactosamine. Microsomal preparations from X163 cells synthesized (14)C-labelled glycosaminoglycan when incubated with UDP-[(14)C]glucuronic acid and either UDP-N-acetylgalactosamine or UDP-N-acetylglucosamine. The (14)C-labelled glycosaminoglycan formed in the presence of UDP-N-acetylgalactosamine was degradable by testicular hyaluronidase, indicating that it was chondroitin-like. The (14)C-labelled glycosaminoglycan formed in the presence of UDP-N-acetylglucosamine was not degradable by testicular hyaluronidase. Microsomal preparations from P815S cells were tested for sulphating activity by incubation with adenosine 3'-phosphate 5'-sulphatophosphate, as well as UDP-[(14)C]glucuronic acid, and UDP-N-acetylgalactosamine. The resulting newly synthesized polysaccharide was shown by chondroitinase ABC digestion to be 70% chondroitin 4-sulphate and 30% chondroitin. The molecular size of this newly synthesized glycosaminoglycan was determined by gel filtration to be larger than 40000 mol.wt. In general, the glycosaminoglycan-synthesizing ability of the microsomal preparations appeared to reflect glycosaminoglycan synthesis by the intact cells.
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PMID:Biosynthesis of glycosoaminoglycans by microsomal preparations from cultured mastocytoma cells. 1674 6

1. Nine acid hydrolases, cytochrome oxidase, alkaline phenylphosphatase and catalase were demonstrated in 0.25m-sucrose homogenates of newborn-rat calvaria. The acid hydrolases were: acid phenylphosphatase, acid beta-glycerophosphatase, beta-glucuronidase, beta-N-acetylglucosaminidase (beta-N-acetylaminodeoxyglucosidase), acid ribonuclease and acid deoxyribonuclease, showing optimum activity at about pH5; cathepsin, beta-galactosidase and hyaluronidase, with optimum activity at about pH3.6. 2. The main kinetic characters of these enzymes have been studied and methods for their quantitative assay have been worked out. The activities present in bone are given and compared with those found in liver. 3. Acid-phosphatase activity was assayed with phenyl phosphate and beta-glycerophosphate as substrates: activities with these two substrates appeared to be due to two different enzymes. Acid phenylphosphatase is particularly labile and is readily inactivated by various physical or chemical agents.
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PMID:Studies on bone enzymes. The assay of acid hydrolases and other enzymes in bone tissue. 1674 42

We report the systematic elaboration of a cost-effective, interference-minimized assay for the label-free optical transduction of chemical reactions. Recently, we have found that certain complexes formed by arginine-rich cell-penetrating peptides (CPPs) and amphiphilic counteranions can act as synergistic anion carriers in lipid bilayer membranes. Application of this discovery to rapid and reversible cytosolic CPP delivery has been described (Futaki, S.; et al. ACS Chem. Biol. 2006, 1, 299). Here, we report the complementary use of polyarginine (pR)-counteranion complexes as general optical transducers of chemical reactions. Counterion screening revealed dodecyl phosphate (DP) as an ideal pR activator. Carboxyfluorescein (CF)-loaded vesicles with a shelf life of 3.5 years served best for the detection of fluorogenic CF release by pR-DP complexes with the naked eye. Inactivation of pR-DP complexes by counterion exchange with hyaluronan (HA) caused no CF emission, while HA removal by hyaluronidase (HAase) did. pR-DP complexes were further compatible with the optical detection of HA immobilization on solid support as well as inhibitor screening for HAase (cromolyn, heparin) with and without substrate immobilization. Controls concerning binary ATP/ADP discrimination for naked-eye kinase detection are mentioned to delineate scope but also limitations of this simple and quite universal method.
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PMID:A cost-effective method for the optical transduction of chemical reactions. Application to hyaluronidase inhibitor screening with polyarginine-counteranion complexes in lipid bilayers. 1698 84

The boundary lubrication function of articular cartilage is mediated in part by proteoglycan 4 (PRG4) molecules, found both in synovial fluid (SF) and bound to the articular cartilage surface. Currently the mechanism by which PRG4 binds to the articular surface is not well understood. The objectives of this study were to determine (1) the effect of bathing fluid contents on PRG4 concentration at the articular surface ([PRG4](cart)), and (2) whether native PRG4 can be removed from the surface and subsequently repleted with PRG4 from synovial fluid. In one experiment, cylindrical cartilage disks were stored in solutions of various PRG4 concentrations, either in phosphate-buffered saline (PBS) or SF as the carrier fluid. In a separate experiment, cartilage disks were stored in solutions expected to remove native PRG4 from the articular surface and allow subsequent repletion with PRG4 from SF. [PRG4](cart) was independent of PRG4 concentration of the bathing fluid, and was similar for both carrier fluids. PRG4 was removed from cartilage by treatment with hyaluronidase, reduction/alkylation, and sodium dodecyl sulphate, and was repleted fully by subsequent bathing in SF. These results suggest that the articular surface is normally saturated with tightly bound PRG4, but this PRG4 can exchange with the PRG4 in SF under certain conditions. This finding suggests that all tissues surrounding the joint cavity that secrete PRG4 into the SF may help to maintain lubrication function at the articular surface.
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PMID:PRG4 exchange between the articular cartilage surface and synovial fluid. 1754 55

The interaction of washed cocci, prepared under specified conditions, and a polynucleotide (AF) results in the formation of streptolysin S provided a fermentable carbohydrate is present. Maximum toxin formation requires, in addition, the presence of magnesium, potassium, and phosphate ions. Streptolysin S production proceeds anaerobically as well as aerobically but under the latter condition, apparently only if the system is sufficiently reducing. Temperature has a marked effect on the rate of appearance of toxin, the critical thermal increment having a value of approximately 36,000. The formation of streptolysin S is inhibited by mercuric ion, arsenite, iodoacetate, dinitrophenol, azide, and other enzyme poisons. The development of streptolysin S in resting cell systems depends neither upon autolysis nor upon physical extraction of preformed toxin but upon toxin synthesis. From the supernatant fluid of the resting cell system, a product containing 20,000 to 30,000 units of streptolysin S per mg. dry weight can be isolated. Information concerning the pH stability of the product is presented. The product is free of streptokinase, hyaluronidase, and proteinase, but possesses appreciable desoxyribonuclease activity. Chemical analyses and other findings indicate that polynucleotide and carbohydrate are present in major amount, and that a small but undetermined quantity of protein is present. Inactivation of streptolysin S by chymotrypsin, ficin, papain, or cathepsin, and not by a variety of other enzymes, indicates that protein is essential for activity, but the precise chemical composition of the toxin remains to be established.
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PMID:Formation of a bacterial toxin (streptolysin S) by resting cells. 1814 84

Previously, we reported the independent tuning of mechanical strength (crosslinking density) and gelation rate of an injectable hydrogel system composed of hyaluronic acid-tyramine (HA-Tyr) conjugates. The hydrogels were formed through the oxidative coupling of tyramines which was catalyzed by hydrogen peroxide (H(2)O(2)) and horseradish peroxidase (HRP). Herein, we studied the encapsulation and release of model proteins using the HA-Tyr hydrogel. It was shown that the rapid gelation achieved by an optimal concentration of HRP could effectively encapsulate the proteins within the hydrogel network and thus prevented the undesired leakage of proteins into the surrounding tissues after injection. Hydrogels with different mechanical strengths were formed by changing the concentration of H(2)O(2) while maintaining the rapid gelation rate. The mechanical strength of the hydrogel controlled the release rate of proteins: stiff hydrogels released proteins slower compared to weak hydrogels. In phosphate buffer saline, alpha-amylase (negatively charged) was released sustainably from the hydrogel. Conversely, the release of lysozyme (positively charged) discontinued after the fourth hour due to electrostatic interactions with HA. In the presence of hyaluronidase, lysozymes were released continuously and completely from the hydrogel due to degradation of the hydrogel network. The activities of the released proteins were mostly retained which suggested that the HA-Tyr hydrogel is a suitable injectable and biodegradable system for the delivery of therapeutic proteins.
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PMID:An injectable hyaluronic acid-tyramine hydrogel system for protein delivery. 1912 48

A novel protocol for the synthesis of biocompatible and degradation controlled poly(lactic-co-glycolic acid) grafted hyaluronic acid (HA-PLGA) was successfully developed for periodontal barrier applications. HA was chemically modified with adipic acid dihydrazide (ADH) in the mixed solvent of water and ethanol, which resulted in a high degree of HA modification up to 85 mol.%. The stability of HA-ADH to enzymatic degradation by hyaluronidase increased with ADH content in HA-ADH. When the ADH content in HA-ADH was higher than 80 mol.%, HA-ADH became soluble in dimethyl sulfoxide and could be grafted to the activated PLGA with N,N'-dicyclohexyl carbodiimide and N-hydroxysuccinimide. The resulting HA-PLGA was used for the preparation of biphasic periodontal barrier membranes in chloroform. According to in vitro hydrolytic degradation tests in phosphate buffered saline, HA-PLGA/PLGA blend film with a weight ratio of 1/2 degraded relatively slowly compared to PLGA film and HA coated PLGA film. Four different samples of a control, OSSIX(TM) membrane, PLGA film, and HA-PLGA/PLGA film were assessed as periodontal barrier membranes for the calvarial critical size bone defects in SD rats. Histological and histomorphometric analyses revealed that HA-PLGA/PLGA film resulted in the most effective bone regeneration compared to other samples with a regenerated bone area of 63.1% covering the bone defect area.
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PMID:Guided bone regeneration by poly(lactic-co-glycolic acid) grafted hyaluronic acid bi-layer films for periodontal barrier applications. 1947 4

Clinically, amniotic membrane (AM) suppresses inflammation, scarring, and angiogenesis. AM contains abundant hyaluronan (HA) but its function in exerting these therapeutic actions remains unclear. Herein, AM was extracted sequentially with buffers A, B, and C, or separately by phosphate-buffered saline (PBS) alone. Agarose gel electrophoresis showed that high molecular weight (HMW) HA (an average of approximately 3000 kDa) was predominantly extracted in isotonic Extract A (70.1 +/- 6.0%) and PBS (37.7 +/- 3.2%). Western blot analysis of these extracts with hyaluronidase digestion or NaOH treatment revealed that HMW HA was covalently linked with the heavy chains (HCs) of inter-alpha-inhibitor (IalphaI) via a NaOH-sensitive bond, likely transferred by the tumor necrosis factor-alpha stimulated gene-6 protein (TSG-6). This HC.HA complex (nHC*HA) could be purified from Extract PBS by two rounds of CsCl/guanidine HCl ultracentrifugation as well as in vitro reconstituted (rcHC*HA) by mixing HMW HA, serum IalphaI, and recombinant TSG-6. Consistent with previous reports, Extract PBS suppressed transforming growth factor-beta1 promoter activation in corneal fibroblasts and induced mac ro phage apoptosis. However, these effects were abolished by hyaluronidase digestion or heat treatment. More importantly, the effects were retained in the nHC*HA or rcHC*HA. These data collectively suggest that the HC*HA complex is the active component in AM responsible in part for clinically observed anti-inflammatory and anti-scarring actions.
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PMID:Biochemical characterization and function of complexes formed by hyaluronan and the heavy chains of inter-alpha-inhibitor (HC*HA) purified from extracts of human amniotic membrane. 1949 Nov 1

Native fluorescence spectroscopy has shown potential to characterize and diagnose oral malignancy. We aim at extending the native fluorescence spectroscopy technique to characterize normal and oral submucous fibrosis (OSF) patients under pre- and post-treated conditions, and verify whether this method could also be considered in the monitoring of therapeutic prognosis noninvasively. In this study, 28 normal subjects and 28 clinically proven cases of OSF in the age group of 20 to 40 years are diagnosed using native fluorescence spectroscopy. The OSF patients are given dexamethasone sodium phosphate and hyaluronidase twice a week for 6 weeks, and the therapeutic response is monitored using fluorescence spectroscopy. The fluorescence emission spectra of normal and OSF cases of both pre- and post-treated conditions are recorded in the wavelength region of 350 to 600 nm at an excitation wavelength of 330 nm. The statistical significance is verified using discriminant analysis. The oxidation-reduction ratio of the tissue is also calculated using the fluorescence emission intensities of flavin adenine dinucleotide and nicotinamide adinine dinucleotide at 530 and 440 nm, respectively, and they are compared with conventional physical clinical examinations. This study suggests that native fluorescence spectroscopy could also be extended to OSF diagnosis and therapeutic prognosis.
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PMID:In vivo native fluorescence spectroscopy and nicotinamide adinine dinucleotide/flavin adenine dinucleotide reduction and oxidation states of oral submucous fibrosis for chemopreventive drug monitoring. 2021 Apr 84


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