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:3.2.1.31 (beta-glucuronidase)
7,680 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Enzymatic digestion with beta-glucuronidase (EC 3.2.1.31) was used to release intact oxazepam from urine samples containing the d5-analog internal standard. The resulting specimens were extracted with Du Pont PREP Type W cartridge (processed by a PREP Automated Sample Processor), Bond Elut Certify, and J.T. Baker "spe" columns for comparison of the columns' extraction recovery and overall effectiveness. Methyl iodide/tetrahexylammonium hydrogen sulfate and N,O-bis(trimethylsilyl)trifluoroacetamide/trimethylchlorosilane (10 g/L) were used for the methylation and trimethylsilylation studies. We used a Hewlett-Packard HP 5790 mass-selective detector equipped with a 13-m J & W DB-5 column (5% phenyl polysiloxane phase) for gas chromatography/mass spectroscopy (GC/MS) analysis and the Thru-Put Target software package for data processing. After several exploratory experiments, we adopted the Du Pont PREP system methylation procedure because of its effective recovery, the superior stability of the derivatization product, the possibility of incorporating a clean-up step, and the potential for high throughput. The extraction recovery from a set of control samples was 87%. Coefficients of variation obtained for six replicates of GC/MS analysis and for the overall procedure were 1% and 3%, respectively. Excellent linearity was established in the 50-8000 micrograms/L concentration range studied. With the use of 3-mL samples, a 20-microL final reconstitution volume, oxazepam at 50 micrograms/L was easily detected under the adopted operation conditions.
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PMID:Enzymatic digestion, solid-phase extraction, and gas chromatography/mass spectrometry of derivatized intact oxazepam in urine. 189 96

Nine healthy volunteers were studied before, during, and after ingesting a fermented dairy product containing Lactobacillus acidophilus, Bifidobacterium bifidum, and mesophilic cultures (Streptococcus lactis and S cremoris) for 3 wk. Hydrogen and methane productions and fecal beta-galactosidase and beta-glucosidase activities were measured as indicators of fermentation capacity of the colonic flora. Fecal concentrations of nitroreductase, azoreductase, and beta-glucuronidase, which may be implicated in colonic carcinogenesis, were also assessed. Hydrogen and methane productions, fecal beta-galactosidase, beta-glucuronidase, and azoreductase activities did not change over three 3-wk periods whereas fecal beta-glucosidase activity increased (42 +/- 6, 91 +/- 12, and 40 +/- 6 IU/g N, P less than 0.01) and nitroreductase decreased (0.87 +/- 0.13, 0.54 +/- 0.11, and 0.57 +/- 0.08 IU/g N, P less than 0.05).
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PMID:Effect of chronic ingestion of a fermented dairy product containing Lactobacillus acidophilus and Bifidobacterium bifidum on metabolic activities of the colonic flora in humans. 211 57

Although the environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), via its interaction with the Ah receptor, is an extremely potent carcinogen and immunosuppressive agent in experimental animals, its possible actions on polymorphonuclear (PMN) function have not been determined. In addition to their importance against infectious organisms, PMNs have been implicated in antitumor resistance. The present studies examined the effects of in vivo exposure to TCDD on PMN function in B6C3F1 (TCDD sensitive, presence of high affinity Ah receptor) and DBA/2N (TCDD resistant at low doses, defective Ah receptor) mice. Animals received a single oral exposure of 5 or 10 micrograms/kg of TCDD and PMNs were obtained 5 days later from the peritoneal cavity following elicitation with sodium caseinate. TCDD reduced the cytolytic and cytostatic activity of PMA-activated PMNs in B6C3F1, but not in DBA/2N mice, suggesting that this response segregates with the Ah locus. Furthermore, TCDD was found to bind specifically to PMNs from Ah-responsive mice. Neither the production of superoxide and hydrogen peroxide nor degranulation, the latter measured by beta-glucuronidase release, was impaired. Supernatants recovered from PMN cell cultures of TCDD-sensitive mice, but not from resistant DBA/2N mice, showed reduced killing capacity for actinomycin D-treated L929 tumor cells, while their ability to bind to tumor cells was not altered. These data suggest that TCDD interferes with PMN-mediated tumor cell killing by altering the production or secretion of a cytolytic factor. Examination of bone marrow stem cells revealed that granulocytic but not monocytic colonies were reduced after TCDD exposure in vivo and in vitro. Although mature PMNs had detectable levels of Ah receptor, exposure in vitro of these cells to TCDD had no effect on antitumor activity. Thus, it is possible that TCDD may affect PMNs at the level of hematopoiesis, via a direct interaction with granulocyte precursor cells, or modulate PMNs at different stages of maturation.
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PMID:Selective inhibition of polymorphonuclear neutrophil activity by 2,3,7,8-tetrachlorodibenzo-p-dioxin. 255 88

Because it has recently been hypothesized that human milk is antiinflammatory, the effects of aqueous human colostrum on human polymorphonuclear leukocyte (PMN) respiratory burst activity and selected enzymatic activities was examined. Aqueous colostrum was found to spontaneously reduce ferricytochrome C in a concentration-dependent manner, prohibiting use of the standard assay to measure superoxide production. It also caused a significant concentration-dependent prolongation of the lagtime from stimulation of PMN with phorbol myristate acetate to the appearance of hydrogen peroxide. Substitution of an enzymatic peroxide-generating system for PMN did not alter the effect of colostrum. Colostrum also suppressed myeloperoxidase activity and lysozyme activity, but not beta-glucuronidase activity in PMN lysates. Inclusion of colostrum in an in vitro assay of PMN-mediated cell detachment significantly suppressed this PMN-mediated effect. These data demonstrate that aqueous human colostrum significantly interferes with PMN oxygen metabolic and enzymatic activities that are important in the mediation of acute inflammation.
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PMID:Antioxidant properties of human colostrum. 284 22

4,4'-Methylenebis(2-chloroaniline) (MBOCA) metabolism in canine liver and kidney slices was investigated using HPLC to separate the metabolites. Liver slices metabolized 5-10% of the 14C-MBOCA in 60 min and produced seven metabolites resolved by HPLC. The major metabolite, representing approximately 80% of the metabolism, was 2-amino-5-[(4-amino-3-chlorophenyl)methyl]-3-chlorophenyl hydrogen sulfate, previously identified as the major urinary metabolite in dogs. An MBOCA-glucoside was identified by mild acid hydrolysis, which released MBOCA and glucose. An O-glucuronide was characterized as labile to beta-glucuronidase, stabile to arylsulfatase, and mild acid. It was formed in increased amounts when 2,6-dichloro-4-nitrophenol (DCNP) was added to the incubation. Two other glucuronide metabolites were labile to mild acid and beta-glucuronidase, stabile to arylsulfatase, and were formed in decreased amounts in the presence of D-(+)-galactosamine (D-gal) and p-nitrophenyl sulfate (PNPS). Renal cortical slices metabolized 3-5% of the 14C-MBOCA in 90 min, producing six metabolites. Based on retention time and lability to hydrolysis, three of these, the MBOCA-glucoside, a glucuronide, and 2-amino-5-[(4-amino-3-chlorophenyl)methyl]-3-chlorophenyl hydrogen sulfate were also found as kidney metabolites. One additional sulfur-containing metabolite was labile to mild acid and arylsulfatase. The major kidney metabolite represented 25-40% of the metabolism and was unaffected by mild acid, beta-glucuronidase, arylsulfatase, DCNP, and D-gal. Covalent binding in liver slices was 20-27 pmol/mg of wet weight/60 min and in kidney was 9-13 pmol/mg of wet weight/90 min.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Metabolism of 4,4'-methylenebis(2-chloroaniline) by canine liver and kidney slices. 287 Aug 90

Basophilic granulocytes were purified from the blood of normal individuals by successive isopyknic centrifugation and elutriation centrifugation. Starting with the leukocyte-rich fraction of 500 ml of blood, we recovered 31 to 80% (mean 51%, n = 20) of the basophils in 45 to 87% purity (mean 69%, n = 23). The contaminating cells were mainly lymphocytes. The basophils were greater than 98% vital (exclusion of ethidium bromide and hydrolysis of fluorescein diacetate). The histamine content of the basophils was 1.1 to 2 pg/cell (mean 1.6 pg/cell, n = 22). With anti-IgE, 30 to 50% of the histamine was released; with phorbol myristic acetate (PMA) or the calcium ionophore A23187, 70 to 100% of the histamine was released. Serum-opsonized zymosan (STZ) did not induce histamine release. Reactions with monoclonal antibodies revealed that the basophils expressed the C3bi receptor (CR3) and the leukocyte function-associated antigen 1 (LFA1), but not the gp 150,95 antigen, the C3b receptor (CR1), or the low avidity Fc gamma receptor. Basophils carry class I but not class II HLA antigens. During incubation of the basophils with serum-opsonized Staphylococcus aureus or Escherichia coli, these bacteria were neither phagocytized nor killed. STZ, PMA, A23187, or anti-IgE did not initiate an "oxidative burst" in the basophils. This was tested with oxygen consumption, cytochrome c reduction, NBT reduction, chemiluminescence, and release of hydrogen peroxide. Moreover, we did not detect cytochrome b558, superoxide dismutase, catalase, or peroxidase in the basophils. Of the typical granule-associated enzymes lysozyme, Vitamin B12-binding protein, and beta-glucuronidase, only beta-glucuronidase was present in the basophils in detectable amounts. This enzyme was released, together with histamine, on incubation of the cells with PMA, A23187, or anti-IgE, but not with STZ. We conclude that basophils from normal human blood are not phagocytes and are probably not involved in the oxidative defense of the host against foreign antigens.
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PMID:Metabolic comparison between basophils and other leukocytes from human blood. 300 19

We describe a liquid chromatographic screening procedure for the detection of stimulant laxatives in urine. A 2-ml urine sample was incubated with 500 U of beta-glucuronidase for 2 h at 60 degrees C. The sample was acidified with sodium acetate (pH 5.0) and extracted with 5 ml of an isopropanol-chloroform (1:9) mixture. The organic layer was cleaned up further by washing with 5 ml disodium hydrogen-phosphate (pH 7.5) before being transferred to a conical tube and evaporated to dryness. The residue was reconstituted in 100 microliters mobile phase and 3 microliters were injected onto a Hewlett-Packard Hypersil ODS (5 microns) column. The ultraviolet absorbance of the eluent was monitored at 225 nm. Rhein, bisacodyl diphenol, bisoxatin diphenol, phenolphthalein, bisacodyl, bisoxatin and danthron all eluted within 6 min. The screen was evaluated using urine specimens obtained from 19 patients who claimed they had taken one or more of the laxatives under consideration within the past 48 h. Only two patients who claimed to have taken Coloxyl and Danthron showed negative results. Eighteen of twenty laxatives (90%) taken by the patients were detected and their identity verified by plotting post-run ultraviolet spectra. We therefore conclude that the screen is sufficiently reliable to be of help in the early detection of surreptitious abusers of stimulant laxatives.
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PMID:Screening procedure for stimulant laxatives in urine using high-performance liquid chromatography with diode array detection. 323 41

The effect of a new prostacyclin analogue OP-41483 on ischemic colonic anastomosis was investigated in dogs. Colonic ischemia was produced by devascularization of the marginal vessels in the left colon and graded into three degrees by measuring colonic blood flow with a hydrogen gas clearance method. The agent was administered intravenously after devascularization using a continuous infusion pump. The parameters studied were colonic blood flow in the submucosal layer, rate of anastomotic leakage, beta-glucuronidase activity and protein content of the colonic mucosa, and histologic changes. After administration of the agent, blood flow increased significantly and beta-glucuronidase activity at the anastomotic site was well preserved at a relatively high level in spite of ischemic change. The anastomotic leakage rate was significantly decreased. The present study proved that administration of this new prostacyclin analogue minimizes ischemic damage, and may be of considerable importance in ischemic colonic anastomoses.
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PMID:Effect of a new prostacyclin analogue on anastomosis of ischemic colon in dogs. 329 62

Receptor-mediated endocytosis and receptor recycling involve a series of intracellular membrane fusion events that appear to play an important role in the regulation of the overall rate and efficiency of the process. An endosome-endosome fusion assay is described using two ligands that (i) rapidly and efficiently enter the endosomal compartment via the macrophage mannose receptor and (ii) that mutually recognize each other. Dinitrophenol-derivatized beta-glucuronidase (DNP-beta-glucuronidase), a ligand for the mannose receptor, was endocytosed by one population of J774 E clone cells, and mannose-derivatized monoclonal anti-DNP IgG (Man-IgG) was internalized by a second set of cells. Both ligands were localized in endosomes as determined by fractionation on Percoll gradients. Incubation of vesicles prepared from the two set of cells resulted in vesicle fusion as indicated by the formation of DNP-beta-glucuronidase-Man-IgG complexes. Under standard conditions, fusion was time-, ATP-, and temperature-dependent. KCl was required for fusion. Fusion required both cytosolic- and membrane-associated proteins. N-Ethylmaleimide treatment of cytosol inhibited fusion. Proton ionophores and amines had no effect on the fusion reaction. ATP-dependent fusion was only observed between early endocytic compartments. While in the presence of a Ca2+ chelator fusion was ATP-dependent, in its absence fusion was also observed in an ATP-independent fashion.
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PMID:In vitro fusion of endosomes following receptor-mediated endocytosis. 336 Jul 75

Tissue circulation of the adrenal gland which is known as antishock organ was investigated by the hydrogen clearance method in hemorrhagic and endotoxin shock using rabbits. Mean blood pressure was maintained at 35 mmHg for two hours in the hemorrhagic group, while 2mg/kg of endotoxin was injected intravenously in the endotoxin group. The data obtained were as follows: Adrenal tissue circulation and renal circulation were reduced to 40% and 14.9%, respectively in the hemorrhagic group and reduced to 69% and 48%, respectively in the endotoxin group. Plasma corticosterone concentration was significantly elevated in both groups. Plasma beta-glucuronidase was significantly elevated in both groups and most animals died within 72 hours. From these data we concluded that adrenal circulation was distributed preferentially at the sacrifice of renal circulation in the shocked state.
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PMID:[Adrenal circulation of rabbits in hemorrhagic and endotoxin shock]. 357 71


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