Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.2.1.31 (beta-glucuronidase)
 7,680 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The aim of the study was the isolation from faecal samples of patients with diarrhoea of verotoxigenic strains of E. coli (VTEC) on the basis of characteristic biochemical properties and production of enterohaemolysin and comparison of isolated verotoxigenic strains with reference strains of VTEC. For isolation of VTEC from 257 stool samples derived from patients with diarrhoea were used selective medium sorbiol--Mac Conkey agar (SMAC) and media supplemented with unwashed and washed in PBS sheep erythrocytes for detection of haemolysins of E. coli. In all haemolytic and sorbitolo-positive or -negative strains isolated from 93 stool samples were examined the activity of beta-glucuronidase using MUG (4-methylumbelliferyl-beta-D-glukuronid) as a substrate for that enzyme. All isolated haemolytic strains as well as reference VTEC were examined on Vero cell line. Verotoxigenic strains from examined samples were investigated by agglutination assay with antiserum to E. coli O157 and then with antisera to eneropathogenic E. coli (EPEC). After that they were examined with ID GN and ATB GN tests. In 93 (36.2%) examined samples there were haemolytic strains of E. coli which fermented or not sorbitol and were MUG-positive or negative. Only in 2 (0.2%) stool samples there were verotoxigenic strains of E. coli which were sorbiol-positive and MG-positive. Both strains belonged to O26 serotype and were derived from samples of two children with diarrhoea. Isolated verotoxigenic strains of E. coli O26 were susceptible on all tested antibiotics.
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PMID:[Characteristic of verotoxigenic strains of Escherichia coli]. 1080 May 77

9-aminocamptothecin glucuronide (9ACG) is a new water-soluble prodrug of 9-aminocamptothecin (9AC) that is a substrate for beta-glucuronidase and displays potent antitumor activity against human tumor xenografts. The lactone ring of camptothecins (CPTs) is required for antitumor activity but spontaneously opens under physiological conditions to an inactive carboxy form. The carboxy form of many CPTs, including 9AC, preferentially binds to human serum albumin (HSA), which further reduces the equilibrium amount of active lactone and greatly decreases antitumor efficacy. In this study, we examined the hypothesis that the unique structure of 9ACG might alter prodrug interaction with HSA and increase 9ACG lactone stability as compared with 9AC. HPLC analysis revealed that HSA did not affect the equilibrium level of 9ACG lactone whereas both CPT lactone and 9AC lactone were greatly reduced in the presence of HSA as compared to their equilibrium levels in PBS. Similar results were found in human serum and whole blood. The lactone ring of 9ACG also opened more slowly (t(1/2)=50 min) as compared with 9AC (t(1/2)=20 min) in the presence of HSA. Both 9ACG lactone and 9ACG carboxy bound HSA with similar affinities (K(D) approximately 4.5 x 10(-5)M(-1)). Binding of 9ACG to HSA reduced prodrug toxicity to cancer cells by about 10-fold in vitro. Injection of HSA into nude mice prolonged the half-life of 9ACG by about 3-fold, indicating that albumin-bound 9ACG lactone may act as a depot of active prodrug in vivo. Our results suggests that in contrast to CPT and 9AC, HSA does not appear to adversely affect 9ACG and may enhance the selective antitumor activity of 9ACG in tumors that contain beta-glucuronidase.
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PMID:Stability of the new prodrug 9-aminocamptothecin glucuronide (9ACG) in the presence of human serum albumin. 1450 97

To increase the therapeutic index of chemotherapeutic drugs, prodrugs have been investigated as anticancer agents, as they may present fewer cytotoxic side effects than conventional cytotoxic drugs, while therapeutic efficacy is maintained or even increased. Extracellular beta-glucuronidase (beta-GUS) in the tumors has been investigated as a target enzyme for prodrug therapy, as it can convert nontoxic prodrugs into cytostatic drugs. To optimize beta-GUS-based prodrug therapies, PET imaging could be a useful tool by providing information regarding the localization and quantification of beta-GUS. Here, we describe our first PET tracer for extracellular beta-GUS, [(18)F]-FEAnGA, which consists of a 2-[(18)F]fluoroethylamine ([(18)F]-FEA) group bound to a glucuronic acid via a self-immolative nitrophenyl spacer. [(18)F]-FEAnGA was synthesized by alkylation of its imidazole carbamate precursor with [(18)F]-FEA, followed by deprotection of the sugar moiety with NaOH in 10-20% overall radiochemical yield. [(18)F]-FEAnGA is about 10-fold more hydrophilic than the cleavage product [(18)F]-FEA, and it is stable in PBS and rat plasma for at least 3 h. In the presence of either Escherichia coli beta-GUS or bovine liver beta-GUS, in vitro cleavage of [(18)F]-FEAnGA with complete release of [(18)F]-FEA was observed within 30 min. C6 glioma cells incubated with the tracer and Escherichia coli beta-GUS or bovine liver beta-GUS showed a 4- and 1.5-fold higher uptake of radioactivity, respectively, as compared to control C6 cells without beta-GUS. Incubation of CT26 murine colon adenocarcinoma cells or the genetically engineered CT26mbetaGUS cells, which expressed membrane-anchored GUS on the outer cell membrane, with the tracer, resulted in a 3-fold higher uptake into GUS-expressing cells as compared to control cells. In a preliminary microPET study in mice bearing both CT26 and CT26mbetaGUS tumors, [(18)F]-FEAnGA exhibited a 2-fold higher retention of radioactivity in the tumor expressing beta-GUS than in the control tumor. [(18)F]-FEA did not show any difference in tracer uptake between tumors. These results suggest that [(18)F]-FEAnGA may be a suitable PET tracer for evaluation of beta-GUS activity, since it is specifically cleaved by beta-GUS and the released [(18)F]-FEA remains attached to targeted cells.
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PMID:Synthesis and evaluation of [18F]-FEAnGA as a PET Tracer for beta-glucuronidase activity. 2041 36

A combination immunoaffinity column (IAC-CZ) clean-up and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analytical method was successfully developed for zearalenol, beta-zearalenol and zearalenone) and chloramphenicol (CAP) in foodstuffs of animal origin. The samples (fish, liver, milk and honey) were enzymatically digested by beta-glucuronidase/sulfatase for about 16 h and then extracted with ether. The extracts were evaporated to dryness and then the residues were dissolved by 1.0 mL of 50% acetonitrile solution. After filtered and diluted with PBS buffer, the reconstituted solution were cleaned-up with a IAC-CZ and then analyzed by LC-MS/MS in multiple reaction monitoring (MRM) mode. The chromatographic separation was performed on a Shimadzu Shim-pack VP-ODS column with gradient elution by acetonitrile and 2 mmol/L ammonium acetate solution. The detection was carried out by electrospray negative ionization mass spectrometry in MRM mode. The proposed method was validated by the limit of detection (0.04-0.10 microg/kg), linearity (R2 > or = 0.999 0), average recoveries (70.9%-95.6%) and precisions (2.0% - 11.8%). The developed method is reliable, sensitive and has good applicability. The combination immunoaffinity column was proved to be an effective pretreatment technique to decrease the matrix effect, and it met the requirements of residue analysis of co-occurring zeranols and chloramphenicol.
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PMID:[Simultaneous determination of zeranols and chloramphenicol in foodstuffs of animal origin by combination immunoaffinity column clean-up and liquid chromatography-tandem mass spectrometry]. 2526 64