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)

Several methods for the preparation of cell suspensions from human gastrointestinal mucosa were investigated. Satisfactory suspensions were obtained by incubating tissue fragments in a solution of collagenase and hyaluronidase overnight at 4 degrees C followed by 30 minutes at 37 degrees C. The resulting suspension contained large numbers of intact lymphoid cells; in addition, variable amounts of epithelial cells and cell debris were present. A high proportion of the lymphoid cells were shown by immunofluorescence to contain immunoglobulin (mainly IgA). Viability of these cells was demonstrated by dye exclusion, their ability to survive in short-term culture, and their ability to incorporate radio-labelled amino acid into immunoglobulin in vitro.
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PMID:Preparation of lymphoid cells from small specimens of human gastrointestinal mucosa. 36 10

Chondrocytes were isolated from adult laryngeal cartilage by an enzymic procedure that included 6 h digestion with collagenase. The level of 35SO4(2-) incorporation into cetylpyridinium chloride-precipitable material by these cells depended upon the subsequent culturing conditions. Suspension cultures incorporated more 35SO4(2-)/cell than monolayer cultures. Hyaluronic acid in the medium inhibited 35SO4(2-) incorporation only when the cells were in primary suspension cultures. It had no effect on monolayer cultures, or monolayers organized into nodules, or suspension cultures derived from monolayers. Mild pretreatment with EDTA, however, rendered these susceptible to hyaluronic acid inhibition. In contrast EDTA abolished the inhibitory effect of hyaluronic acid on primary suspension cultures. Oligosaccharides, derived from hyaluronidase digestion of hyaluronic acid that were larger than decassaccharide, had some inhibitor effect on 35SO4(2-) incorporation by monolayer cultures. The total 35SO4(2-) incorporation was less in primary suspension cultures of chondrocytes isolated after 12 h than after 6 h digestion of cartilage and the inhibition by hyaluronic acid was also less. These differences persisted during 12 days of culture. It is suggested that the method of isolating chondrocytes and subsequent culture conditions may modify the cell surface and mask or abolish specific binding sites for hyaluronic acid.
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PMID:Synthesis of proteoglycans by suspension and monolayer cultures of adult chondrocytes and de novo cartilage nodules-the effect of hyaluronic acid. 41 57

Computerized image-analysis techniques have been employed to examine the sarcomere dynamics of isolated mammalian cardiac myocytes. The cells were prepared by perfusion of adult rabbit hearts with hyaluronidase-collagenase solutions; they exhibited phasic contractions in the presence of 10(-6) M Ca2+. The dissociated cells were visualized by phase microscopy and a video camera interfaced in a minicomputer. Digitized cell images were processed by an algorithm utilizing signal averaging and contrast enhancement to yield data showing individual sarcomere position and shortening vs. time, so that patterns of sarcomere activation could be observed in spontaneously contracting cells. Compared to records of whole-cell shortening and of striation displacement, computerized image analysis provided a much more faithful indication of time course and sequence of sarcomere shortening. Spontaneously contracting cells showed sequential sarcomere shortening beginning at one end and propagating longitudinally with a constant velocity, typically at 100--150 micron/s for beat rates of 40 min-1. Velocities of initial sarcomere shortening appeared to increase with elevated Ca2+. These observations are consistent with a regenerative mechanism of calcium-induced calcium release.
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PMID:Sarcomere motion in isolated cardiac cells. 43 41

Soluble 125I-labeled type I collagen binds to cultured fibroblasts but not to cultured epithelia. The binding of the ligand to fibroblasts is reversible, saturable and highly specific for sequences contained within the helical portions of the alpha1 and alpha2 chains. The amount of ligand bound is dependent upon cell number and ligand concentration. Binding is decreased but measurable at 4 degrees C. The steady state binding is greater at 26 degrees than at 37 degrees C due to a more rapid dissociation of the ligand-acceptor complex at 37 degrees C. The half-life of the complex is 46 min at 37 degrees C and approximately 2.5 hr at 26 degrees C. Scatchard plots of binding data indicate a single class of high affinity binding sites (KD = 1.2 X 10(-11) M) with each fibroblast binding approximately 500,000 molecules at saturation. Pretreatment of fibroblasts with bacterial collagenase, chondroitinase ABC or testicular hyaluronidase does not affect the binding reaction, whereas pretreatment of the cells with phospholipase C increases the amount of ligand bound. Ligand binding is decreased but not abolished after fibroblasts are treated with trypsin concentrations which remove surface fibronectin. Fibroblast monolayers treated with antiserum against fibronectin bind the radiolabeled ligand normally. In contrast to collagen, addition of excess fibronectin does not accelerate the dissociation of bound ligand from fibroblasts. Possible functions for surface-bound collagen are discussed.
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PMID:Binding of soluble type I collagen molecules to the fibroblast plasma membrane. 45 36

Erythrocytes of different species (chicken, sheep, man, mouse, rat, guinea pig) except rabbit erythrocytes strongly adhere to the marginal zone of mouse spleen follicles in frozen sections. This adherence reaction (AR) is not restricted to red blood cells but is also observed with human lymphocytes. Pretreatment of the tissue sections with trypsin, mercaptoethanol, periodate, chloroform/methanol, acetone, and heating the sections abolishes AR whereas neuraminidase (VCN) treatment of the sections has an amplifying effect. AR is inhibited by preincubation of the neuraminidase- or untreated sections with neuraminic acid (NA). Treatment of the erythrocytes with VCN completely abolishes AR whereas treatment with other enzymes (hyaluronidase, collagenase) is ineffective in this respect. Determination of NA in the erythrocyte membrane before and after VCN treatment reveals a positive correlation between the amount of NA and AR. Rabbit red blood cells have the lowest NA content in their membranes and, in addition, there is little effect of VCN treatment in further reducing it. It is possible that a lectin-like substance is responsible for AR. The biologic significance of AR is hypothetical, but since AR occurs in an area of the spleen playing a role in antigen trapping it is conceivable that this trapping may be mediated by interaction of NA and NA receptor(s).
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PMID:Erythrocyte adherence to the marginal zone of mouse spleen follicle mediated by receptor(s) for neuraminic acid. 46 83

The bulk isolation from rat cerebral cortex of viable neurons retaining synaptic complexes is described. The basis of this procedure is to dissociate the neurons in situ from the surrounding glial cells. The glial structures that are normally adjacent to the neuronal cell body and to the proximal parts of the neuronal processes are largely destroyed by perfusion of the brain under special conditions. The most important of these conditions was found to be a hyperosmolar concentration of hexoses in the perfusion medium. In addition, the presence of collagenase and hyaluronidase in the perfusion medium and specific perfusate flow characteristics were required to produce the structural changes throughout the brain tissue. When the perfused brain was further dissociated into a cell suspension by mincing and sieving, isolated neurons were obtained, the majority of which retained the proximal parts of their processes. A novel feature of these neurons was the retention of synaptic boutons on the plasma membrane. Presynaptic terminals with mitochondria and vesicles as well as pre- and postsynaptic membranes and densities were observed on the isolated neurons. The neurons were fractionated to 90--95% purity using discontinuous Ficoll density gradient centrifugation with a liquid fluorocarbon as cushion. Highly purified, viable cerebral neurons retaining synaptic complexes are thus available in bulk for neurobiological studies.
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PMID:Neurochemical and morphological studies of bulk isolated rat brain cells. II. Preparation of viable cerebral neurons which retain synaptic complexes. 46 41

During the first 10 days after peripheral deafferentation of the mouse olfactory bulb stereoselective binding of L-[3H]carnosine declines markedly. The initial phase of this decline is due to a decrease in binding site stereoselectivity, which is then followed by a loss of assayable binding sites. The specificity of inhibition of L-[3H]carnosine binding by various peptides is also altered after denervation. Competitive inhibitors of carnosine binding become less potent after denervation, while analogues which are not competitive inhibitors remain equipotent before and after denervation. Several carnosine analogues that are normally poor inhibitors become more potent after denervation. Treatment of bulb membranes with trypsin, RNase and hyaluronidase, but not DNase or collagenase, resulted in significant alterations in carnosine binding. L-, but not D-carnosine, protected the binding site from trypsin digestion, and induced additional binding in bulb membranes in a dose-and temperature-dependent fashion. Preincubation of membranes with L-carnosine also led to the induction of additional carnosine binding in membranes from cerebral cortex, cerebellum and deafferentated bulbs but not from muscle. Bulbs from newborn mice contain about one-half of the adult levels of binding and no significant sex differences in carnosine binding were detected in bulbs from adult rats. L-[3H]carnosine binding was two-fold higher in the anterior compared to the posterior portion of the bulb, but there were no significant differences in binding of opiate, GABA, alpha-adrenergic, muscarinic cholinergic, benzodiazepine of glutamic acid receptor ligands.
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PMID:L-[3H]Carnosine binding in the olfactory bulb. II. Biochemical and biological studies. 48 25

Sertoli cells have been insolated from the newborn calf testis using a combination of mechanical and enzymatic disruption. Testicular fragments, previously chopped into 1-mm pieces, are digested in an enzyme mixture consisting of hyaluronidase, collagenase, trypsin and DNAse, followed by a second digestion in trypsin and DNAse. Isolation of the resulting cellular fractions by sedimentation with unit gravity produces an aliquot of Sertoli cells which is over 95% pure when examined by light and electron microscopy. Cultures of these cells grow rapidly and produce Mullerian Inhibiting Substance as evidenced by their ability to cause the involution of the Mullerian duct of the female fetal rat when co-cultured in an organ-culture assay system.
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PMID:The secretion of Mullerian inhibiting substance by cultured isolated Sertoli cells of the neonatal calf. 51 48

1. A method for the preparation of isolated mammary gland cells of the rat is described. 2. The procedure involves disaggregation of the tissue in a collagenase-hyaluronidase mixture and subsequent purification of the heterogeneous population of cells by centrifugation in discontinuous Ficoll-400 gradients; the preparation takes 60 minutes. The yield of cells is approximately 14%. 3. The cells as prepared have high rates of metabolism and synthetic capacity and exhibit metabolic characteristics comparable to intact tissue. 4. Measurements of the content of metabolic intermediates show cells to have, and retain, outstandingly high levels of ATP and to have an energy charge close to 0.9. Levels of other intermediates approximate to those found in the intact tissue. The level of glycolytic intermediates below the triose phosphate stage indicate the highly aerobic state of the cells. 5. The pattern and scale of glucose utilization, measured using specifically labelled glucose incorporation into 14CO2 and 14C-labelled lipid production, approximates closely in isolated cells at 5 and 20 mM glucose and in tissue slices at 20 mM glucose concentration. Mammary gland slices incubated with 5 mM glucose have a considerably lower rate of metabolism. Isolated cells exhibit a higher proportionate rate of glucose utilization by way of the pentose phosphate pathway. 6. The isolated cells are hormone responsive. Insulin increases the oxidation of glucose by the pentose phosphate pathway and stimulates lipid synthesis. Addition of progesterone and cortisone in vitro (10 muM) leads to a marked and rapid decrease in the rate of glucose oxidation and conversion to lipid.
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PMID:Regulation of mammary gland metabolism: pathways of glucose utilization, metabolite profile and hormone response of a modified mammary gland cell preparation. 67 49

Inapparent nodule-transformed cells were recovered from five late-pregnant, first-pregnancy BALB/cfC3H females 4 months of age and from five late-pregnant multiparous females 6 to 7 months of age. Mammary tissues were removed from each donor and dissociated by means of the enzyme collagenase (0.1%), hyaluronidase (0.1%), and pronase (1.25%). Aliquots of 100,000 viable cells in 0.01 ml of media were injected into the gland-free mammary fat pads of 3-week-old syngeneic host mice. Ten weeks after the injection the outgrowths were classified as ductal, nodule, tumor, or combinations of these types of outgrowths. The recovery of nodule outgrowths indicated the presence of nodule-transformed cells in the cell suspension that was injected. All donors yielded nodule outgrowths, and the percentage of outgrowths was significantly greater than was the percentage recovered from virgin BALB/cfC3H females of the same age groups. The latent period for the emergence of nodules and tumors was reduced from 8 to 9 months in virgin females to 4 months in parous females. The incidence of both nodules and tumors was greatly increased. The data suggest that parity significantly increases the numbers of nodule-transformed cells in donor tissue, decreases the time required for the emergence of nodules and tumors, and increases the number of overt nodules and tumors.
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PMID:Effect of parity on recovery of inapparent nodule-transformed mammary gland cells in vivo. 69 53


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