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:3.1.21.1 (DNase)
7,655 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Spleen lymphocytes of BCG-immunized mice contain a soluble factor that inhibits in vitro the growth of the H37Rv strain of Mycobacterium tuberculosis within normal peritoneal macrophages. The water-soluble extracts of sensitized lymphocytes, disrupted by freezing and thawing, although less active than the corresponding viable cells retained a significant growth-inhibiting activity. Dialysis against distilled water, lyophilization, exposure to ribonuclease and deoxyribonuclease, and storage at -20 degrees C of the water-soluble extracts did not affect their antimycobacterial activity, whereas extracts heated at 100 degrees C were completely devoid of such an activity. All the inhibiting activity was recovered in the void volume of the column after chromatography on Sephadex G-200. Water-soluble constitutents of sensitized lymphocytes did not affect BCG grown in vitro, and on repeated treatments of tuberculous mice they led to a negligible protection against pulmonary tuberculosis. Preliminary observations seem to indicate that other soluble factors in lymphocytes of BCG-sensitized mice have the capacity to potentiate in vitro the phagocytic activity of normal macrophages.
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PMID:Partial characterization of a factor extracted from sensitized lymphocytes that inhibits the growth of Mycobacterium tuberculosis within macrophages in vitro. 82 9

Hyaluronic acid was prepared from adult rabbit skin. Defatted skin powder suspended in 0.5 M NaCl was homogenized, and total glycosaminoglycans were precipitated from this 0.5 M NaCl extract with cetylpyridinium chloride, then redissolved successively with increasing concentrations of NaCl and finally 0.5 N NaOH. Hyaluronic acid, the major acid glycosaminoglycan in the 0.5 M NaCl extract, was purified and fractionated by DEAE-Sephadex chromatography. The molecular weights ranged from 1 X 10(4) to 7.2 X 10(4). Alternatively, hyaluronic acid was obtained from adult rabbit skin without mechanical powdering and homogenizing. Defatted skin pieces were suspended in water and heated at 100 degrees, then the extract was digested with pronase followed by DNase [EC 3.1.4.5]. Glycosaminoglycans were excluded in gel filtration with Sephadex G-75. Hyaluronic acid and dermatan sulfate, the two major glycosaminoglycans of this tissue, were separated by gel chromatography on Sepharose 4B. The molecular weight of this hyaluronic acid ranged from 1.6 X 10(5) to 1.3 X 10(6). Yields of hyaluronic acid by these two methods were similar. Hyaluronic acid was probably degraded by the mechanical treatments in the first method. Other factors affecting the viscosity of the tissue extract were examined.
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PMID:Molecular weight of hyaluronic acid from rabbit skin. 84 39

O antigen extracted from whole cells of Bacteroides fragilis ss. fragilis NCTC 9343 with 45 per cent aqueous phenol has been purified by gel filtration and chromatography. First, the water phase was treated with RNase and DNase and passed through a column of agarose. The chromatographic procedures included ion exchange on a column of DEAE-cellulose and adsorption to hydroxylapatite. The O antigen was eluted from the DEAE-cellulose with a gradient of NaCl, and from the column of hydroxylapatite with 1 M phosphate buffer, pH 6.8. Inhibition of indirect haemagglutination was used to detect the O antigen in the eluates.
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PMID:Purification of the O antigen of Bacteroides fragilis ss. fragilis NCTC 9343 from phenol-water extracts by gel filtration and chromatography on deae-cellulose and hydroxylapatite. 96 34

The bone inducing factor derived from BF osteosarcoma was purified in the following manner. Step 1. The sarcoma, grown in CBA mice, was excised and lyophilized. Step 2. The powder was washed with chilled acetone. Step 3. The acetone-treated powder was then homogenized with chilled distilled water. Step 4. Washing with 0.15M KCl. Step 5. The precipitate was incubated in in 0.2 N NH2OH, pH7.0, for 48 H at 25 degrees. After Step 5, the bone-forming activity showed a slight increase; however, the factor remained insoluble. The properties of the factor were as follows. The factor is relatively relatively heat stable; the osteogenic activity survived the treatment at 75 degrees for 15 min or at 55 degrees for 19 h. The activity was easily lost by mechanical shaking. Incubation with DNase, RNase, neuraminidase, chondroitinase ABC and beta-galactosidase left the osteogenic activity intact, but treatment with either pronase or collagnease destroyed this activity. The results suggest that the factor may be a protein. The activity was seen with the lyophilized BF osteosarcoma cells (without matrix), and it is probable that the factor was exclusively synthesized in the cells. The bone formation, observed across a millipore filter when living BF osteosarcoma enclosed in a millipore chamber was implanted in mice, suggests the synthesis and secretion of the factor from the cells.
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PMID:Studies on a factor responsible for new bone formation from osteosarcoma in mice. 105 58

During bacteriophage studies on Haemophilus influenzer, it was observed that encapsulated type b and unencapsulated Rb strains released a bactericidal substance acitve against types a, c, d, e, and f H. influenzae, non-typable H. influenzae strains, other Haemophilus species, and certain members of the Enterobacteriaceae. The bactericidal activity was assayed by a plaque test utilizing an Rd strain as an indicator lawn and was also demonstrated in mixed broth cultures of a producer strain and an indicator strain. Immediately lysis of sensitive bacteria by the factor was not evident. The factor is sensitive to trypsin but resistant to deoxyribonuclease, treatment with 2-mercaptoethanol, lipase, alpha-amylase, and heating in a 100 degrees C water bath for 20 min. The activity is not dependent upon increased Ca2+ or Mg2+ concentration as is necessary for HP1C1 and S2 phage propagation. The bactericidal factor is not pelleted by high-speed centrifugation at 150,000 X g for 6 h. Treatment with ultraviolet light or mitomycin C does not result in observable phage, phage-like particles, or increased bactericidal activity. T-HE BACTERICIDAL FACTOR IS NOT A TYPICAL SMALL MOLECULAR WEIGHT "COLICIN-LIKE" BACTERiocin in that it is not inducible, has a wider range of activity, and does not kill by "single-hit" kinetics. On preliminary characterization, it is a thermostable protein toxic to certain bacterial strains.
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PMID:Bactericidal substance produced by Haemophilus influenzae b. 108 28

Experiments were carried out to assess the survival of recombinant plasmid DNA during large-scale production of recombinant human erythropoietin (rhuEPO) in a fermentation pilot plant. The analyses revealed DNA-degrading activities in the fermentation broth and in the waste-water, leading to rapid destruction of plasmid DNA added to medium or waste-water. The capability of the plasmid-DNA-spiked samples to transform competent bacteria was drastically reduced. The DNA-degrading activity in the waste-waters could be blocked by addition of EDTA or by boiling, indicating the presence of DNA-degrading enzymes (DNases). No plasmid-specific DNA sequences were detected in waste-water samples by in-vitro amplification with Taq-polymerase. Genomic DNA preparations of cell debris collected from waste-water samples only contained degraded plasmid DNA. Furthermore, it was shown that intact plasmid DNA could be degraded to fragments of less than 1000 bp by incubation at 121 degrees C for 20 min, leading to a decrease in the plasmid-specific transforming capacity by a factor of 10(3) per minute. Thus, DNA from the rhuEPO production pilot plant was efficiently inactivated at three different levels: (i) in the fermentation medium (DNase), (ii) in the waste-water container (DNase), and (iii) by heat inactivation for 20 min at 120 degrees C. These results indicate that the probability of delivery of recombinant DNA into the environment is extremely low in such biotechnological production processes.
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PMID:Inactivation of recombinant plasmid DNA from a human erythropoietin-producing mouse cell line grown on a large scale. 136 78

For simultaneous cytophotometric measurement of DNA and RNA, the standardized Methyl Green-Pyronin Y technique is an obvious choice. It is, however, first necessary to correlate the uptake of Pyronin Y to the staining intensity of RNA. The material consisted of paraffin sections of formalin- or Carnoy-fixed rat liver. The sections were pretreated with water, buffer, deoxyribonuclease, ribonuclease, or both enzymes in sequence, and stained with the standardized Methyl Green-Pyronin Y procedure, Gallocyanin chromalum, or the Feulgen reaction. Sections stained directly without pretreatment served as controls. Staining intensities were measured with an image analyser for cell nuclei, nucleoli and cytoplasm. After deoxyribonuclease treatment, nuclear staining intensity with Methyl Green, Gallocyanin chromalum, and Schiff's reagent dropped nearly to zero. The same was seen for both nucleoli and cytoplasm with Pyronin Y and Gallocyanin chromalum after ribonuclease treatment. Staining intensity of Pyronin Y correlated directly with that of Gallocyanin chromalum for nucleoli and cytoplasm. After ribonuclease treatment, a direct correlation was found between the nuclear staining intensity of Methyl Green and nuclear absorption of Gallocyanin chromalum. We conclude that the standardized Methyl Green-Pyronin Y stain is reliable for the simultaneous quantitative assessment of both RNA and DNA. The simplicity of this technique makes it a valuable tool even for daily routine.
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PMID:Simultaneous quantification of DNA and RNA in tissue sections. A comparative analysis of the methyl green-pyronin technique with the gallocyanin chromalum and Feulgen procedures using image cytometry. 137 24

Serum levels of IgM, IgG and IgG-antibody subclasses directed against cell envelopes, lipopolysaccharides and cytoplasmic fractions from Capnocytophaga sputigena, C. gingivalis and C. ochracea were examined in age-, race- and sex-matched periodontally healthy (n = 25) subjects and subjects with adult periodontitis (n = 25). The envelopes and cytoplasmic fractions were obtained by ballistic disintegration of the cells and ultracentrifugation. Cell envelopes were treated with DNase, RNase and lysozyme. Lipopolysaccharides were obtained by hot phenol-water extraction and treated with DNase and RNase. The relative levels of the antibodies in response to the cell fractions were measured by the streptavidinbiotin micro enzyme-linked immunosorbent assay. Both groups showed IgM and IgG antibodies to each fraction of the three Capnocytophaga species, but the frequency of positive IgG subclass responses varied. The IgG4 responses were lower than the other subclasses. There were no significant differences between the IgM antibody levels of the two groups. However, the adult periodontitis group had significantly lower IgG antibody titres to the cell envelopes and cytoplasmic fractions of C. gingivalis and C. ochracea, and lipopolysaccharide of C. gingivalis. These results were reflected in the depressed levels of IgG1 and/or IgG2 to these cellular fractions from the same bacterial species. The adult periodontitis group also showed a lower level of IgG1 to the cytoplasmic fractions of C. sputigena without any depression in the total IgG antibody level. There were no significant differences between the groups in IgG3 and IgG4 antibody levels to any of the cellular fractions.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Serum antibody responses in human periodontitis to cellular components of Capnocytophaga. 141 21

Enterococcus faecalis 226 NWC, isolated from natural whey cultures utilized as starter in water-buffalo Mozzarella cheese manufacture, produces a bacteriocin, designated Enterocin 226 NWC, which is inhibitory to Listeria monocytogenes. Plasmid analysis of E. faecalis 226 NWC showed a single 5.2-kb plasmid, pEF226. In conjugation experiments, pEF226 was transferred into a plasmid-free strain of E. faecalis JH2-2. The transfer required direct cell-to-cell contact and was not inhibited by DNase. The identity of conjugation was confirmed by digestion with SmaI restriction endonuclease and subsequent pulsed-field gel electrophoresis (PFGE) of the genomic DNA of E. faecalis 226, E. faecalis JH2-2 and of the isolates after the mating. The data indicate that the ability of E. faecalis 226 NWC to produce the bacteriocin is linked to the 5.2-kb conjugative plasmid pEF226.
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PMID:Conjugal transfer of plasmid-borne bacteriocin production in Enterococcus faecalis 226 NWC. 146 12

The glomerular basement membrane (GBM) is an integral structural component of the glomerular filter, but its contribution to the hydraulic and macromolecular permeability properties of the glomerulus has been the subject of much controversy. We have modified previously reported methods to develop a technique with which to study filtration properties of microgram quantities of isolated GBM in vitro at physiological pressures. Rat glomeruli were sieved, and cells were removed with N-laurylsarcosine and DNase. GBM (150 micrograms; greater than 95% pure) were added to a mini-ultrafiltration cell and consolidated under pressure to form a continuous filter at the base of the cell. Water flux was identical to inulin clearance at applied pressures less than 150 mmHg and increased with progressive increments in the transmembrane pressure. Hydraulic conductivity of GBM was inversely related to the prevailing transmembrane pressure gradient. The hydraulic conductivity depended on albumin concentration in a manner that was not monotonic, with the conductivity being lower at 4 g/dl albumin than at 0 or 8 g/dl. When plasma was utilized as the retentate, the fractional clearance of albumin was over twice that of immunoglobulin G, and the fractional clearance of each protein was much higher than that in the intact glomerulus. On the basis of these results, both the hydraulic and macromolecular permeability of an individual layer of GBM are much greater than that reported for the intact glomerulus. The large quantitative differences between GBM permeability and that of intact glomeruli suggest a major contribution of cellular elements to glomerular permeability properties.
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PMID:Glomerular basement membrane: in vitro studies of water and protein permeability. 162 16


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