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)

Two rapid methods were evaluated for their extraction of plasmids from Clostridium perfringens. The first method involved lysis of 1 to 2 ml of C. perfringens culture by treatment with hyaluronidase, lysozyme, and sarcosyl. DNA, extracted with phenol-chloroform, was treated with RNase, boiled, and electrophoresed in a 1.2% agarose gel. The second method involved lysis of 2 ml of culture by lysozyme treatment and extraction with alkaline sodium dodecyl sulfate (SDS). Extracted DNA was treated with RNase, boiled, and electrophoresed in a 0.7% agarose gel. Of 57 strains of C. perfringens analyzed by both extraction procedures, 11 were shown to have plasmids by the alkaline SDS method which were missed by the phenol-chloroform extraction method. These new plasmids were of higher molecular mass and ranged up to 68 megadaltons. Use of the DNase inhibitor diethyl pyrocarbonate did not further improve the yield of plasmid DNA. An additional 159 isolates of C. perfringens screened by the alkaline SDS method revealed plasmids up to 80 megadaltons in mass and an overall plasmid carriage rate of 69%.
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PMID:Rapid extraction of plasmids from Clostridium perfringens. 287 Jun 80

Antigens extracted from cells of Streptococcus pyogenes T6 and Streptococcus mutans strains AHT, BHT, 10449, OMZ175, and K1R adsorbed to the sarcolemmal sheath of cardiac muscle cells in vitro. Similar preparations from S. salivarius, S. sanguis, Staphylococcus aureus, and Lactobacillus casei had weak or negligible tissue-binding activity. Tissue-bound bacterial antigens were detected with homologous rabbit antisera with both indirect immunofluorescence tests and an indirect radioimmunoassay. Serological cross-reactivity was observed between the tissue-binding factors of S. pyogenes and S. mutans cells but not between the bacteria and muscle tissue. In a comparative study of extraction procedures, the greatest yield of tissue-binding factors was obtained from group A streptococci by cell disruption in buffer at 4 degrees C. Hot aqueous phenol and hot water extracts were inactive. Antibodies specific for the tissue-binding factor(s) were readily adsorbed from rabbit anti-S. pyogenes serum by a preparation of isolated cytoplasmic membranes but not by a suspension of cell wall fragments. The heart-binding component of S. pyogenes cell extracts was inactivated by protease digestion and heat treatment and to a lesser extent by periodic acid oxidation. The capacity of heart cell components to adsorb streptococcal antigens was reduced by protease treatment but not by the action of neuraminidase, hyaluronidase, organic solvents, or detergents.
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PMID:Binding of streptococcal antigens to muscle tissue in vitro. 699 20

Myocardial cells were isolated after treatment with collagenase (0.05%) and hyaluronidase (0.1%) by discontinuous-gradient centrifugation on 3% Ficoll. Nuclei derived from these myocardial cells were then fractionated on a discontinuous sucrose density gradient with the following steps: (I) 2.0M/2.3M, (II) 2.3M/2.4M, (III) 2.4M/2.5M, (IV) 2.5M/2.6M, and (V) 2.6M/2.85M. The myocardial nuclei were sedimented in the interfaces of gradient fractions (II) and (III). Nuclei from whole ventricles that had been treated with the enzymes before isolation sedimented into five major subsets of nuclei. These findings suggest that nuclei sedimented in the isopycnic gradient at fractions (II) and (III) are most probably derived from myocardial cells. However, this procedure is laborious and lengthy, and the recovery of myocardial-cell nuclei is low. An alternative method was developed to isolate an enriched fraction of myocardial-cell nuclei from whole ventricular tissue without exposing the tissues to enzyme digestion. These ventricular nuclei could be fractionated into five nuclear subsets by using the same discontinuous sucrose density gradient as that described above. The content of DNA, RNA and protein per nucleus for each band was determined. Although the DNA content per nucleus was constant (10pg), that of RNA varied from 1.5 to 4.5pg and that of protein from 16 to 24pg. Nuclei from each band were examined by light-microscopy: large nuclei occurred in the ligher regions whereas smaller nuclei were found in the denser regions of the gradient. From the size distribution pattern of myocardial-cell nuclei compared with that of total ventricular nuclei, it was found that nuclear subsets (II), (III), and (IV) were similar to myocardial nuclei. Electrophoretic analyses of the proteins solubilized in sodium dodecyl sulphate/phenol or Tris/EDTA/2-mercaptoethanol/phenol obtained from each nuclear subset indicate that these fractions are similar, with limited qualitative differences. These findings indicate that isolation of an enriched fraction of myocardial-cell nuclei could be achieved by discontinuous-sucrose-density-gradient centrifugation.
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PMID:Fractionation of rat ventricular nuclei. 739 68

Using the death of mice due to the growth of tumor homografts as an end point, the distribution and properties of a tumor homograft-promoting substance(s), the "enhancing substance," were studied. Activity was most largely present in tumor and spleen tissues while that in the liver, kidney, and stomach was relatively slight. Activity was widely distributed in fractions prepared from tumor. Although DNAP and crude RNAP fractions were active, activity could not be associated with either RNA or DNA. The fraction possessing the highest concentration of activity was a residue fraction which was insoluble in water or in dilute or strong saline solutions. Extraction of lyophilized tumor with acetone or 3:1 alcohol:ether reduced, but did not eliminate, activity in the insoluble fraction. However, activity was completely lost after extraction with 80 per cent alcohol at room temperature. Enhancing activity of a tumor fraction was irreversibly lost after exposure for 1 hour to a solution more acid than pH 5 or more alkaline than pH 9. Activity was completely lost after exposure to 50 per cent urea or 90 per cent phenol. Activity was not altered by incubation with trypsin, hyaluronidase, DNAase or the receptor-destroying enzyme of V. cholerae, but it was decreased when trypsin or hyaluronidase was injected with the incubated tumor fraction. Activity was rapidly destroyed by treatment with dilute solutions of sodium periodate at pH 7. Under similar conditions, treatment with KMnO(4) resulted in less extensive destruction of activity while H(2)O(2), K(2)Cr(2)O(7), and NaIO(4) previously inactivated by reaction with glucose had no apparent effect on activity. These results are interpreted as indicating the presence of both carbohydrate and protein in the structure of the activity substance.
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PMID:Studies on a substance that promotes tumor homograft survival (the enhancing substance); its distribution and some properties. 1340 73