Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:6.3.4.6 (urease)
 7,490 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Intra-strain variation in the expression of lipopolysaccharide (LPS) by two clinical isolates of Helicobacter pylori was examined. Lipopolysaccharide was prepared from successive cultures of individual colonies from each strain, separated by SDS-PAGE, and detected by silver staining and by immunoblotting. The genetic 'relatedness' of the colonies was investigated using PCR-RFLP analysis of the urease and vacuolating cytotoxin genes. Although individual colonies of each of the two strains examined appeared to have the same genetic origins, variation in the expression of their long-chain LPS was observed. The same LPS profiles were maintained by individual colonies over four subcultures on solid media containing 10% (v/v) defibrinated horse blood.
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PMID:Intra-strain variation in expression of lipopolysaccharide by Helicobacter pylori. 971 8

A potentiometric urea-sensitive biosensor using a NH4(+)-sensitive disposable electrode in double matrix membrane (DMM) technology as transducer is described. The ion-sensitive polymer matrix membrane was formed in the presence of an additional electrochemical inert filter paper matrix to improve the reproducibility in sensor production. The electrodes were prepared from one-side silver-coated filter paper, which is encapsulated for insulation by a heat-sealing film. A defined volume of the NH4(+)-sensitive polymer matrix membrane cocktail was deposited on this filter paper. To obtain the urea-biosensor a layer of urease was cast onto the ion-sensitive membrane. Poly (carbamoylsulfonate) hydrogel, produced from a hydrophilic polyurethane prepolymer blocked with bisulfite, served as immobilisation material. The disposable urea sensitive electrode was combined with a disposable Ag/AgCl reference electrode to obtain the disposable urea biosensor. The sensor responded rapidly and in a stable manner to changes in urea concentrations between 7.2 x 10(-5) and 2.1 x 10(-2)mol/l. The detection limit was 2 x 10(-5) mol/l urea and the slope in the linear range 52 mV/decade. By taking into consideration the influence of the interfering K(+)- and Na(+)-ions the sensor can be used for the determination of urea in human blood and serum samples (diluted or undiluted). A good correlation was found with the data obtained by the spectrophotometric routine method.
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PMID:A disposable biosensor for urea determination in blood based on an ammonium-sensitive transducer. 1002 47

Cell surface proteins of the human gastric pathogen Helicobacter pylori extracted during different in vitro growth phases were analyzed by one- and two-dimensional gelelectrophoresis (1-DE and 2-DE) and by 2-DE immunoblot. Broth-cultured H. pylori cells were stained with an acridine-orange dye to monitor the morphological status of the organism. In 2-day-cultures, 96% of the bacterial cells were spiral-shaped and four days later a morphological switch to coccoid forms occurred. In 10-day cultures spiral-shaped forms were not found. By 1-DE, proteins with the molecular masses of 87 and 120 kDa were detected in the 2-day cultures that disappeared in cells of 12-day cultures. A protein corresponding in size to the heat shock protein (GroEl homolog, Hsp60) and a 62 kDa protein, the ureaseB-subunit, were identified in extracted proteins of 2-, 8-, and 12-day cultures. 2-DE revealed an increased number of silver-stained spots of 8-day cultures (in average 250 spots) compared with protein extracted from 2-day cells (in average 160 spots). 2-DE immunoblots performed with sera containing antibodies to major H. pylori proteins such as the A- and B-subunits of urease and the Hsp60 showed similar reactivity to surface proteins extracted from 2-, 8-, and 12-day cultures, suggesting that these proteins remain immunologically intact. Pooled sera from infected patients absorbed with spiral-shaped cells showed an almost total blocking of the antibody reactivity to extracted coccoid proteins in 2-DE immunoblot. Eighteen spots were still visible, but this reactivity probably represents a solid overexpression by the coccoid cells of Hsp60 and ureaseB proteins and is thus difficult to block.
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PMID:Two-dimensional electrophoretic and immunoblot analysis of cell surface proteins of spiral-shaped and coccoid forms of Helicobacter pylori. 1094 44

We compared immunohistochemical and silver stains of pediatric gastric biopsy sections for the identification of Helicobacter pylori infection with chronic inflammation and a negative urease screening test. Thirty-seven patients (age range 10 months to 21 years) whose gastric antral biopsies were negative for the rapid urease test (CLO(R)) but positive for lymphocytic infiltration were selected for a retrospective study. Specimens had been subjected to a rapid urease test (CLO(R)) and hematoxylin and eosin staining, and Dieterle silver staining and immunohistochemical staining specific for H. pylori were also performed. Twelve additional patients with urease-positive biopsies were used as controls. With Dieterle staining, 8/37 (22%) urease-negative biopsies contained organisms morphologically compatible with H. pylori, 21/37 (56%) contained organisms not compatible with H. pylori, and 8/37 (22%) were negative for organisms. Immunostaining confirmed 6/8 (75%) Dieterle-positive cases as being H. pylori, was negative in 2/8 (25%) Dieterle-positive cases, and was positive in 2/8 (25%) Dieterle-negative cases. Biopsies from 8/12 (67%) urease-positive specimens contained organisms seen with both Dieterle and immunohistochemical stains, and 4/12 (33%) were negative with both stains. Although both stains yielded comparable results with H. pylori-positive biopsies, Dieterle staining was potentially confusing because of nonspecific staining of other organisms. A significant proportion of (CLO(R))-negative biopsies was positive for H. pylori with special stains. We therefore recommend the use of immunohistochemical staining rather than silver staining in the evaluation of urease-negative gastric biopsies demonstrating chronic inflammation in children.
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PMID:Comparison of immunohistochemistry and silver stain for the diagnosis of pediatric Helicobacter pylori infection in urease-negative gastric biopsies. 1120 Apr 95

Detailed histopathological evaluation of the gastric mucosa of Helicobacter-infected cats is complicated by the difficulty of recognizing Helicobacter organisms on hematoxylin and eosin (HE)-stained sections and the ability of multiple Helicobacter species to infect cats. In this study, the presence and localization of different species of Helicobacter in the stomachs of cats was investigated using silver staining and immunohistochemistry. Five groups containing 5 cats each were established (group 1: urease negative and Helicobacter free; groups 2, 3, 4, and 5: urease positive and infected with Helicobacter heilmannii, unclassified Helicobacter spp., Helicobacter felis, and Helicobacter pylori, respectively). Gastric samples were evaluated by HE and silver staining and by immunohistochemistry with 3 different anti-Helicobacter primary antibodies. Helicobacter were detected by Steiner stain in all infected cats at the mucosal surface, in the lumen of gastric glands, and in the cytoplasm of parietal cells. In silver-stained sections, H. pylori was easily differentiated from H. felis, H. heilmannii, and unclassified Helicobacter spp., which were larger and more tightly coiled. No organisms were seen in uninfected cats. Helicobacter antigen paralleled the distribution of organisms observed in Steiner-stained sections for 2 of the 3 primary antibodies tested. The antisera were not able to discriminate between the different Helicobacter species examined. A small amount of Helicobacter antigen was present in the lamina propria of 3 H. pylori-, 3 H. felis-, and 1 H. heilmannii-infected cat. Minimal mononuclear inflammation was present in uninfected cats and in those infected with unclassified Helicobacter spp. and H. heilmannii cats. In H. felis-infected cats, lymphoid follicular hyperplasia with mild pangastric mononuclear inflammation and eosinophilic infiltrates were present. The H. pylori-infected cats had severe lymphoid follicular hyperplasia and mild to moderate mononuclear inflammation accompanied by the presence of neutrophils and eosinophils. These findings indicate that Steiner staining and immunohistochemistry are useful for detecting Helicobacter infections, particularly when different Helicobacter species can be present. Monoclonal antibodies specific for the different Helicobacter species could be important diagnostic aids. There appear to be differences in the severity of gastritis in cats infected with different Helicobacter species.
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PMID:Histological and immunohistochemical detection of different Helicobacter species in the gastric mucosa of cats. 1124 59

Four palladium(II) aqua complexes catalyze hydrolytic decomposition of urea into carbon dioxide and ammonia. The initial rates of carbon dioxide formation at 313 K and pH 3.3 fall in the range 6.7 x 10(-)(5) to 1.6 x 10(-)(4) M min(-)(1), depending on the catalyst. The pseudo-first-order rate constant for the formation of carbon dioxide is 1.7 x 10(-)(3) min(-)(1) in the presence of 0.30 M cis-[Pd(en)(H(2)O)(2)](2+) as the catalyst at 313 K and pH 3.3. This reaction is ca. 1 x 10(5) times faster than the uncatalyzed decomposition of urea. The reaction catalyzed by cis-[Pd(en)(H(2)O)(2)](2+) is monitored by (13)C and (15)N NMR spectroscopic methods. The following steps in the mechanism of this reaction are studied quantitatively: binding of urea to the catalyst, formation of carbamic acid (H(2)NCOOH) coordinated to palladium(II) via the nitrogen atom, and conversion of this intermediate into carbon dioxide and ammonia. These products are formed also by another pathway that does not involve carbamic acid. Kinetic effects of added acid and inhibition of the reaction by addition of thiourea and of bases are interpreted quantitatively. Ammonia inhibits the decomposition. When, however, this product is sequestered by metal cations, the reaction becomes relatively fast and catalytic turnover is achieved. The most effective of these sequestering agents is the silver(I) cation. Although the simple palladium(II) complexes are very different from the enzyme urease, which contains nickel(II) ions, the decomposition of urea catalyzed by both kinds of agents involves carbamic acid as the intermediate. Kinetic and mechanistic studies with metal complexes contribute to the understanding of the enzymatic mechanism.
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PMID:Kinetics and Mechanism of Urea Hydrolysis Catalyzed by Palladium(II) Complexes. 1167 Feb 15

The palladium(II) aqua complex cis-[Pd(en)(H(2)O)(2)](2+) catalyzes the alcoholysis of urea into alkyl carbamate and ammonia. The observed rate constants for the ester formation fall in the range from 1.8 x 10(-)(5) to 5.9 x 10(-)(1) min(-)(1) at 313 K and pH 3.3, depending on the alcohol. This catalyzed reaction is at least 10(5) times faster than the uncatalyzed alcoholysis of urea under the same conditions. This is the first example of catalytic, nonhydrolytic cleavage of the amide bond in urea. The following steps in the mechanism of the methanolysis reaction are studied quantitatively: binding of urea to the catalyst in the presence of various alcohols or various concentrations of water, direct methanolysis of O-bound and N-bound urea, formation of carbamic acid (NH(2)COOH) coordinated to palladium(II) via the nitrogen atom, methanolysis of this intermediate, and the fast dissociation resulting in free methyl carbamate. Ammonia, a product of alcoholysis, inhibits this reaction by binding to palladium(II). When, however, ammonia is sequestered by the silver(I) cation, alcoholysis becomes relatively fast, and catalytic turnover is achieved. Various alcohols are compared in their reactivity toward urea. The effects of nucleophilicity, steric bulk, size, and additional hydroxyl groups (in diols) are examined. The intramolecular alcoholysis in the 2,6-dithia-1,8-octanediol complex cis-[Pd(C(6)H(14)O(2)S(2))(H(2)O)(2)](2+) results in at least 100-fold rate enhancement relative to the intermolecular alcoholysis by cis-[Pd(en)(H(2)O)(2)](2+). Alkyl carbamates do not hydrolyze further into carbamic acid and alcohol. Aryl carbamates do hydrolyze further, and this reaction requires the palladium(II) aqua complex as a catalyst. Carbamic acid then spontaneously decomposes into carbon dioxide and ammonia. Observed rate constants for the appearance and disappearance of aryl carbamates agree with the relative nucleophilicities of aryl alcohols. This study of the catalysis by a metal complex may contribute to the understanding of the metalloenzyme urease. We propose a new method, alcoholysis, for cleaving amide bonds in peptides and proteins.
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PMID:Alcoholysis of Urea Catalyzed by Palladium(II) Complexes. 1167 May 66

A novel helicobacter with the proposed name Helicobacter cetorum, sp. nov. (type strain MIT 99-5656; GenBank accession number AF 292378), was cultured from the main stomach of two wild, stranded Atlantic white-sided dolphins (Lagenorhynchus acutus) and from the feces of three captive cetaceans (a Pacific white-sided dolphin [Lagenorhynchus obliquidens]; an Atlantic bottlenose dolphin [Tursiops truncatus]; and a beluga whale [Delphinapterus leucas]). The infected captive cetaceans were either subclinical, or clinical signs included intermittent regurgitation, inappetance, weight loss, and lethargy. Ulcers were observed in the esophagus and forestomach during endoscopic examination in two of the three captive animals. In the third animal, esophageal linear erosions were visualized endoscopically, and histopathological evaluation of the main stomach revealed multifocal lymphoplasmacytic gastritis with silver-stained spiral-shaped bacteria. Helicobacter cetorum is a fusiform gram-negative bacterium with a single bipolar flagellum. The isolates grow under microaerobic conditions at 37 and 42 degrees C but not at 25 degrees C. H. cetorum is urease, catalase, and oxidase positive, and it is sensitive to cephalothin. The isolates from the wild, stranded dolphins were sensitive to nalidixic acid, whereas the isolates from the collection animals were resistant. By 16S rRNA sequencing it was determined that H. cetorum represented a distinct taxon that clusters most closely with H. pylori. Further studies are necessary to determine the role of H. cetorum in the development of gastric ulcers and gastritis of cetaceans. This is the first description and formal naming of a novel Helicobacter species from a marine mammal.
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PMID:Helicobacter cetorum sp. nov., a urease-positive Helicobacter species isolated from dolphins and whales. 1245 48

The relationship between urinary infections and stone formation has been recognized since antiquity and it has been over a century since bacterial degradation of urea was postulated to cause struvite stones. Specific therapy for urease-producing bacteria, such as urease-inhibitors and antibiotics, has allowed for treatment for this subset of urinary stones. Future directions for research include development of novel urease-inhibitors and chemicals to enhance the protective glycosaminoglycan layer. An improved understanding of the pathogenesis of calcium-based stones has led to the discovery of potential roles for nanobacteria and Oxalobacter formingenes. Methods of altering intestinal regulation of oxalate by reintroduction of lactic acid bacteria may significantly impact the treatment of calcium oxalate stones. The use of catheters, both urethral and ureteral, is common in the urinary tract and is associated with significant morbidity, primarily from associated infections. Catheters to prevent bacterial colonization and formation of biofilms have been created using various coatings, including ciprofloxacin, hydrogel, and silver. Use of these types of catheters may minimize infections and encrustation inherent with their placement in the urinary tract.
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PMID:Infections and urinary stone disease. 1267 63

Subclinical gastritis was observed in 10 of 10 baboons (Papio spp.) from a toxicity study in a research facility. The lesions were similar in xenobiotic-treated and control animals, suggesting a spontaneous rather than chemical-induced disease. Histologic examination revealed lymphoplasmacytic gastritis in the antral mucosa. The fundic mucosa contained minor, scattered aggregates of lymphocytes and plasma cells. A Warthin-Starry silver stain and ultrastructural examination revealed numerous spiral-shaped bacteria morphologically resembling Helicobacter pylori in antral glands and numerous spiral-shaped bacteria morphologically consistent with H. heilmannii-like organisms in fundic glands. Polymerase chain reaction assay of paraffin-embedded antral and fundic tissue using primers for the urease gene and 16S ribosomal ribonucleic acid gene amplified deoxyribonucleic acid fragments with a high degree of sequence homology for H. pylori and H. heilmannii. This is the first report of gastritis associated with Helicobacter-like organisms in baboons.
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PMID:Gastritis associated with Helicobacter-like organisms in baboons. 1294 14


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