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)

Large numbers of Pneumocystis carinii (2 X 10(10) nuclei) were isolated and separated from the lungs of immunosuppressed rats by an enzymatic (collagenase, hyaluronidase and DNase) digestion procedure. The nucleic acid isolated from this P. carinii-enriched preparation was characterized by melting point analysis and RNA-sizing gels. The GC content of P. carinii DNA was approximately 33% while the rat DNA was 41.4%. In addition, RNA isolated from the P. carinii-enriched preparation showed unique ribosomal RNA bands of 3.4 kb and 1.8 kb as compared with uninfected rat lung ribosomal RNA which banded at 4.8 and 1.9 kb. Following isolation and fragmentation by sonication, the P. carinii DNA fragments were inserted into the vector, lambda gt-11. The resultant library contained 1.1 X 10(5) phage, of which 40-45% hybridized to P. carinii DNA but not to rat DNA.
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PMID:Characterization and cloning of Pneumocystis carinii nucleic acid. 252 83

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

We have recently provided evidence from studies conducted in vivo that the ovary, particularly by means of estrogen, regulates placental androstenedione (delta 4A) production during the second half of rat pregnancy. In the present study, an incubation system of dispersed rat placental cells was established to determine if estrogen acts directly on the placenta to regulate delta 4A production. Placentas were obtained on Days 14-15 of rat gestation and dispersed in Hanks' Balanced Salt Solution containing 0.1% collagenase, 0.1% hyaluronidase, 0.01% DNase, and 1% fetal calf serum. Placental cells were incubated in Medium 199 for 16 h at 37 degrees C. A time-dependent increase (r = 0.96, p less than 0.05) in the release of delta 4A occurred over the 16-h incubation period. Mean +/- SE formation of the steroid intermediate progesterone (P4) and product delta 4A was 1.17 +/- 0.78 and 1.18 +/- 0.22 ng per 10(7) cells respectively. The addition of 1-10 microM diethylstilbestrol (DES) decreased (p less than 0.05-0.01) delta 4A production, and had no significant effect on P4 or pregnenolone (P5) formation. The percent decrease in delta 4A production was 14.2 +/- 12.9, 30.9 +/- 2.3, and 55.0 +/- 4.4 with 1, 5, and 10 microM DES, respectively. Treatment of placental cells with estradiol (E2) also resulted in a decrease (p less than 0.01) in delta 4A production with no effect on P4 formation. The percent inhibition of delta 4A production was 34.2 +/- 11.1 and 77.3 +/- 5.2 with the addition of 1 microM and 10 microM E2, respectively. E2 (10 microM) produced a concomitant threefold increase (p less than 0.01) in P5 formation.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:The effect of estrogen on androstenedione production by rat placental cells in vitro. 292 52

Using monoclonal antibodies, we examined the display of rabbit Ia by articular chondrocytes. We found that 29 to 46% of chondrocytes displayed Ia antigen compared with 46 to 60% of spleen cells. Ia antigen expression was not likely to be the result of enzyme treatment. To investigate antigen presenting activity of enzyme dissociated normal articular chondrocytes, adult rabbits were immunized in the front foot pads with ovalbumin (OVA) in complete Freund's adjuvant. Four to six weeks later, draining popliteal lymph node cells (LNC) were obtained. Articular chondrocytes were obtained by overnight collagenase, DNase, and hyaluronidase digestion of cartilage from both ends of femurs and proximal end of tibias. Antigen-presenting cells from spleen were used as positive controls. LNC and nylon wool-purified T cells were cultured with OVA pulsed and mitomycin C-treated chondrocytes or spleen cells, and lymphocyte proliferation was measured by 3H-TdR uptake. Both chondrocytes and spleen cells showed antigen presenting activity, and stimulation of lymphocyte proliferation was inhibited by murine monoclonal anti-rabbit Ia antibody (2C4), whereas control plasmacytoma cell supernatants had no effect. When T cells were purified first by Sephadex G-10 and later by nylon wool columns, these cells were dependent on antigen-presenting cells for immunogen (OVA)-induced lymphocyte proliferation. Again, chondrocytes under these strict experimental conditions presented antigen to T cells. Chondrocytes also stimulated autologous and allogeneic normal lymphocytes. Thus, normal chondrocytes have Ia antigens on their surface and can function as antigen-presenting cells. These results are significant for the understanding of local cellular interaction in the pathogenesis of rheumatoid arthritis.
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PMID:Class II histocompatibility antigen-mediated immunologic function of normal articular chondrocytes. 293 76

A total of 237 breast carcinomas have been studied with the Courtenay-Mills (C-M) soft agar method. Cell yields and plating efficiencies (PE) were recorded after various enzyme treatments. The highest cell yields and PEs were obtained with the combination of collagenase 0.5%, hyaluronidase 1000 IE ml-1 and DNase 0.1% and an incubation time of 2 h. Eighty percent of the specimens gave greater than 10 colonies, and 60% formed greater than 30 colonies permitting chemosensitivity studies. The C-M method gave significantly higher PEs than the Hamburger-Salmon (H-S) method. Hormone supplements (insulin, oestradiol, progesterone, hydrocortisone) and also reduced agar concentrations (less than 0.3%) gave marginal stimulation of colony formation. In chemosensitivity studies involving doxorubicin, vincristine and 4-OOH-cyclophosphamide, the C-M method gave dose-response relationships without plateaus.
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PMID:Cultivation of human breast carcinoma in soft agar. Experience with 237 fresh tumour specimens. 304 54

The glycosaminoglycans that exist in rabbit bone marrow were analyzed chemically, and their in situ localization was studied immunohistochemically. Femoral bone marrow of 3-month-old rabbits was defatted with organic solvents. Glycosaminoglycans were prepared from the defatted tissue after its digestion with pronase, treatment with mild alkali, and then digestion with DNase-I. The tissue contained glycosaminoglycans equivalent to 195 mg of hexosamine per femur, which accounted for 27.3% of the total hexosamine in the tissue. Studies with hyaluronidase from Streptomyces hyalurolyticus and chondroitinase ABC showed that the glycosaminoglycans were composed of hyaluronic acid (16% of the total glycosaminoglycan) and chondroitin 6-sulfate (79%). The chondroitin 6-sulfate was separated on Bio-Gel A-0.5m gel into two molecular species with mol wt of greater than 12,000 (Kd greater than 0.2) and approximately 8,000 (Kd = 0.47). Bone marrow digested with chondroitinase ABC and then treated with three monoclonal antibodies 4/8/9-A-2, 5/6/3-B-3, and 5/6/1-B-5, which were specific for unsaturated 4-sulfated, 6-sulfated, and nonsulfated disaccharide structures, respectively, at the nonreducing end of chondroitin sulfate chains, reacted with only 5/6/3-B-3. This result indicated that the chondroitin sulfate, isomer in the bone marrow is chondroitin 6-sulfate, consistent with the biochemical results. The chondroitin 6-sulfate was localized mainly in the extracellular compartment and was considered to be involved in construction of the hemopoietic microenvironment in the bone marrow.
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PMID:Isolation, characterization, and localization of glycosaminoglycans in rabbit bone marrow. 311 61

Thirty-three strains of Vibrio vulnificus of clinical and environmental origin were examined for production of 12 extracellular enzymes of potential importance to the virulence of this bacterium. Strains of Vibrio vulnificus were consistent in their production of protease, mucinase, lipase, chondroitinase, hyaluronidase, DNase, sulfatase, and hemolysin. No differences between clinical and environmental isolates were noted. Although none of the enzymes appeared to correlate with the ability of these strains to produce lethality in mice, the production of hemolysin and of a protease with activity against native serum albumin may be significant in the pathogenesis of the potentially fatal infections produced by this organism. The production of several of these exoenzymes also appeared to correlate with pathogenicity in the seven other Vibrio species examined. Culture filtrates of all virulent strains of Vibrio vulnificus were cytotoxic for Chinese hamster ovary cells, whereas those of the strains of Vibrio parahaemolyticus and Vibrio alginolyticus examined lacked this activity.
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PMID:Production of extracellular enzymes and cytotoxicity by Vibrio vulnificus. 352 90

Cell suspensions were prepared by either mechanical or enzymatic disaggregation methods from biopsy specimens from 54 patients with various tumors. The biologic activities of cells derived from the two suspensions were then examined. Biopsy specimens of solid tumors were minced, and one-half of each specimen was further processed by being teased with needles, whereas the other half was exposed to a combination of collagenase, hyaluronidase, and DNase. The enzymatic disaggregation method yielded fewer cells per gram of tissue than the mechanical method. However, the percentage of dye-excluding cells was increased by the enzymatic procedure in 93% of the cases. Cells obtained by enzymatic means also had higher cloning efficiencies than those obtained by mincing. The histologic types of cells present in the initial cell suspensions were the same for cells obtained by the enzymatic or mechanical disaggregation methods. The number of colonies obtained was linearly related to the number of cells plated in both cases. The tritiated thymidine suicide indices (estimates of the percentage of cells in the S-phase of the cell cycle) were the same for the two cell populations obtained by the two methods. The results indicate that cells obtained from solid tumors by enzymatic dissociation methods did not differ significantly from cells obtained by the more conventional mechanical techniques. However, cell viabilities and cloning efficiencies were significantly improved by the enzymatic technique.
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PMID:Effect of enzymatic disaggregation on proliferation of human tumor cells in soft agar. 628 27

Production of monodispersed cell suspensions from primary human breast tumors is difficult due to the predominant stromal composition of most breast tumors. Our studies were designed to optimize dispersion of breast tumors of known stromal content and histopathology. In a first series of experiments three enzymatic protocols were compared to disperse minced tissue: (A) treatment with collagenase (2 mg/ml) in the presence of 5% serum for 24 hours; (B) treatment with collagenase (6 mg/ml) and DNase (0.002%) in 10% serum for 3 hours; (C) treatment with collagenase (2 mg/ml) for 3 hours followed by pronase (0.075%) for 1 hour. Protocol A produced better cell yields than B or C for all tumors tested. The monodispersed cells were suspended in a 0.3% semi-solid agar with alpha modified Eagles medium (alpha MEM), 10% serum, and selected hormones, then layered over similarly enriched 0.5% semi-solid agar. The cells prepared by protocol A had a higher plating efficiency and larger average colony size than B or C. In a second series of experiments, protocol A was repeated and compared to two additional procedures: (D) treatment with collagenase (2 mg/ml) and hyaluronidase (1 mg/ml) in the presence of 5% serum for 24 hours; and (E) mechanical disaggregation. Protocol D exhibited a small but significant negative difference from A, while E was the least efficient in producing viable monodispersed cells from the tumors. All enzymatically monodispersed cells produced clonal growth in our agar system. However, mechanically dispersed cells gave growth in only 4 of 7 tumors. Protocol A, in addition to yielding the highest number of viable cells per gram of tissue, gave the highest plating efficiency of all protocols tested.
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PMID:A comparison of methods for the production of monodispersed cell suspensions from human primary breast carcinomas. 630 35

Advanced human periodontitis is considered to be a B-cell lesion, but the cellular infiltrate contains several cell types, the distribution of which has not been determined. This experiment was designed to characterize and identify the immunocompetent cells on histological sections and in eluates from diseased human gingiva. Immunoglobulin-bearing cells were detected on histological sections by direct immunofluorescence with F(ab')2 antisera monospecific for human immunoglobulin G (IgG), IgA, or IgM. Plasma cells predominated in the central portion of the lamina propria, with the proportions positive for IgG, IgA, and IgM accounting for 65.2 +/- 9.5, 11.2 +/- 1.1, and 1.3 +/- 1.1% of the total infiltrating cells, respectively. T lymphocytes, identified by indirect immunofluorescence with monoclonal antibody (Leu-1) against human T cells, accounted for 29.3 +/- 10.0% of the total infiltrated cells. Most of the T cells were located subjacent to the pocket epithelium, but there were a few in the central lamina propria. Similarly, Fc receptor-bearing cells detected by EA rosetting and macrophages and monocytes detected by nonspecific esterase staining with alpha-naphthylbutyrate esterase were also localized to the region immediately subjacent to the pocket epithelium. Infiltrated cells were harvested from minced gingival tissue after digestion with collagenase, hyaluronidase, and DNase. The eluates contained 35.3 +/- 6.0% T lymphocytes, 30.0 +/- 14.9% Fc receptor-bearing cells, and 12.9 +/- 4.4% monocytes and macrophages. Whereas T gamma cells comprised 13.3 +/- 1.4% of peripheral blood T cells, they accounted for only 6.0 +/- 2.0% of the eluate T cells. In contrast, T mu cells accounted for 44.7 +/- 4.9% of the T cells in the eluates and 51.6 +/- 4.4% in the peripheral blood. The decreased proportion of T gamma cells in the gingiva may indicate a form of abnormal immune regulation concerned with T suppression of B-cell proliferation.
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PMID:Identification and distribution of immunocompetent cells in inflamed gingiva of human chronic periodontitis. 660 70


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