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)

The role of the periplasmic alpha-carbonic anhydrase (alpha-CA) (HP1186) in acid acclimation of Helicobacter pylori was investigated. Urease and urea influx through UreI have been shown to be essential for gastric colonization and for acid survival in vitro. Intrabacterial urease generation of NH3 has a major role in regulation of periplasmic pH and inner membrane potential under acidic conditions, allowing adequate bioenergetics for survival and growth. Since alpha-CA catalyzes the conversion of CO2 to HCO3-, the role of CO2 in periplasmic buffering was studied using an alpha-CA deletion mutant and the CA inhibitor acetazolamide. Western analysis confirmed that alpha-CA was bound to the inner membrane. Immunoblots and PCR confirmed the absence of the enzyme and the gene in the alpha-CA knockout. In the mutant or in the presence of acetazolamide, there was an approximately 3 log10 decrease in acid survival. In acid, absence of alpha-CA activity decreased membrane integrity, as observed using membrane-permeant and -impermeant fluorescent DNA dyes. The increase in membrane potential and cytoplasmic buffering following urea addition to wild-type organisms in acid was absent in the alpha-CA knockout mutant and in the presence of acetazolamide, although UreI and urease remained fully functional. At low pH, the elevation of cytoplasmic and periplasmic pH with urea was abolished in the absence of alpha-CA activity. Hence, buffering of the periplasm to a pH consistent with viability depends not only on NH3 efflux from the cytoplasm but also on the conversion of CO2, produced by urease, to HCO3- by the periplasmic alpha-CA.
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PMID:The periplasmic alpha-carbonic anhydrase activity of Helicobacter pylori is essential for acid acclimation. 1562 43

Improvements to the efficiency of dietary nitrogen use by lactating dairy cattle can be made by altering the concentration and form of protein in the diet. This study collected urine and feces from dairy cows from selected crude protein (CP) treatments of 2 lactation studies. In the first trial, collections were made from cattle fed a diet with high (19.4%) or low (13.6%) CP content (HCP and LCP, respectively). In the second trial, collections were made from cattle fed diets in which the forage legume component was alfalfa (ALF) or birdsfoot trefoil with a low (BFTL) or high (BFTH) concentration of condensed tannins (CT). A system of small laboratory chambers was used to measure NH3 emissions over 48 h from applications of equal quantities of urine and feces to cement (simulating a barn floor) and from applications of slurries, made by combining feces and urine in the proportions in which they were excreted for each treatment, to soil. Reducing dietary CP content resulted in less total N excretion and a smaller proportion of the excreted N being present in urine; urine N concentration was 90% greater for HCP than LCP. Surprisingly, NH3 emissions from the barn floor were similar in absolute terms despite the great differences in urine urea-N concentrations, presumably because urease activity was limiting. Cumulative emissions from fresh slurries applied to soil represented 18% of applied N for both HCP and LCP. Following storage at 20 degrees C for 2 wk, cumulative emissions from LCP were much lower than for HCP, representing 9 and 25% of applied N, respectively. Emissions were also lower when expressed as a proportion of slurry total ammoniacal N (TAN) content (24 and 31%, respectively) because of treatment differences in slurry pH. Increasing CT content of the dietary forage legume component resulted in a shift in N excretion from urine to feces. Cumulative NH3 emissions from the barn floor were greater for ALF than for BFTL or BFTH. Emissions from fresh and stored slurries were in proportion to slurry TAN contents, with approximately 35% of applied TAN being lost for all treatments. Emissions expressed as a proportion of total N applied were consistently lower for BFTH than for ALF.
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PMID:Dietary manipulation in dairy cattle: laboratory experiments to assess the influence on ammonia emissions. 1582 70

The localization of the activity of immobilized urease on chitosan membrane was studied by X-ray microanalysis. BaCl2 and urea were selected as the capture and substrate respectively. The substrate was hydrolyzed by immobilized urease to produce NH3 and CO2 in Tris-HCl buffer (pH 7.0), and the latter was captured by BaCl2 to form precipitate. The precipite was deposited on the active site of immobilized urease. It is shown that the method is practicable and reliable. The optimum condition for the localization of activity of immobilized urease was studied.
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PMID:[X-ray microanalysis of the activity of immobilized urease on chitosan membrane]. 1601 32

Tight junctions, paracellular permeability barriers that define epithelial cell polarity, play an essential role in transepithelial transport, cell-cell adhesion and lymphocyte transmigration. They are also important for the maintenance of innate immune defence and intestinal antigen uptake. Ammonium (NH4+) is elevated in the gastric aspirates of Helicobacter pylori-infected patients and has been implicated in the disruption of tight-junction functional integrity and the induction of gastric mucosal damage during H. pylori infection. The precise mechanism of the effect of ammonium and the molecular targets of ammonium in host tissue are not yet identified. To study the effects of ammonium on epithelial tight junctions, the human colon carcinoma cell line Caco-2 was cultured on permeable supports and the transepithelial resistance (TER) was measured at different time intervals following exposure to ammonium salts or H. pylori-derived ammonium. A biphasic response to treatment with ammonium was found. Acute exposure to ammonium salts or NH3/NH4+ derived from urea metabolism by wild-type H. pylori resulted in a 20-30 % decrease in TER. After 24 h, the NH4Cl-treated cells showed a partial recovery of TER. In contrast, the control culture, or cultures that were exposed to supernatants derived from urease-deficient H. pylori, showed no significant decrease in TER. Occludin-specific immunoblots revealed the expression of a low-molecular-weight form of occludin of 42 kDa upon NH3/NH4+ exposure. The results indicate that modulation of tight-junction function by H. pylori is ammonium-dependent and linked to the accumulation of a low-molecular-weight and detergent-soluble form of occludin.
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PMID:Production of ammonium by Helicobacter pylori mediates occludin processing and disruption of tight junctions in Caco-2 cells. 1620 10

The aim is the assessment of the HP infection in stomach using breath test and comparison to other diagnostic methods, as well as following up the effect of therapy. In 83 patients with digestive discomfort rapid urease test, histology and breath test were performed, while in 25 patients with proven HP infection the effect of therapy was followed up using breath test and clinical findings. For rapid urease test and histology, samples were taken from antral mucosa. Breath test was performed after per oral administration of the capsule of 14C- urea (37 kBq) (Izotop, Hungary and Laboratory for radioactive isotopes, Vinca) which, in the presence of Helicobacter pylori breaks up to 14CO2 and NH3. Radioactivity was measured by beta counter in the exhaled air fasting and 30 minutes after ingestion of the capsule. According to our results, the rise of activity over 100% was considered positive. From 83 patients, 58 were breath test was positive, 24 negative and one equivocal. Fast urease test was in 54 positive, in 29 negative while histology was in 57 postitive and 26 negative. Findings of the breath and urease tests were in accordance in 93% patients while breath test and histology in 98% patients. During follow up of the therapeutic effects, breath test and clinical findings were in accordance in 98% patients. Breath test can be useful in diagnosis but is a method of choice in following up the patients after therapy for H. pylori infection, because it is non-invasive, fast and precise.
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PMID:[Radionuclide detection of Helicobacter pylori infection]. 1640 32

A novel determination method for urea using an acid urease column-FIA system was developed, and the system was applied to the determination of urea in rice wine. This novel FIA system was characterized by CO2 detection due to the property of acid urease and by a microfluidic gas-diffusion device with the use of an ultra-thin hollow fiber membrane. A biosensing system fabricated in this study was assembled with a double-plunger pump, a sample-injection valve, an immobilized acid urease column as a recognition element for the assay of urea, a gas-diffusion unit, and a flow-type spectrophotometer. The gas-diffusion unit consisted of a double-tubing structure in which the outer tubing was made of PTFE (i.d. 1.0 mm; o.d. 1.5 mm) and the inner tubing was of porous PTFE (i.d. 0.19 mm; o.d. 0.25 mm). Standard urea solutions (20 microl) were measured through monitoring variations in the absorbance of a coloring agent solution resulting from a pH shift due to carbon dioxide molecules being enzymatically generated. A wide and linear relationship was obtained between the concentration of urea (16 microM - 1.0 mM) and the change in absorbance. This FIA system has great advantages that the system did not suffer from ammonia and ethanol in samples. This system, armed with a microfluidic gas-diffusion device, was applicable to the determination of various substrates of many kinds of decarboxylase, amino-acid oxidase, and amino-acid oxygenase, producing CO2 and NH3 molecules.
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PMID:Novel determination system for urea in alcoholic beverages by using an FIA system with an acid urease column. 1642 98

Due to its low digestibility in the small intestine, a major fraction of the polyol isomalt reaches the colon. However, little is known about effects on the intestinal microflora. During two 4-week periods in a double-blind, placebo-controlled, cross-over design, nineteen healthy volunteers consumed a controlled basal diet enriched with either 30 g isomalt or 30 g sucrose daily. Stools were collected at the end of each test phase and various microbiological and luminal markers were analysed. Fermentation characteristics of isomalt were also investigated in vitro. Microbiological analyses of faecal samples indicated a shift of the gut flora towards an increase of bifidobacteria following consumption of the isomalt diet compared with the sucrose diet (P<0.05). During the isomalt phase, the activity of bacterial beta-glucosidase decreased (P<0.05) whereas beta-glucuronidase, sulfatase, nitroreductase and urease remained unchanged. Faecal polyamines were not different between test periods with the exception of cadaverine, which showed a trend towards a lower concentration following isomalt (P=0.055). Faecal SCFA, lactate, bile acids, neutral sterols, N, NH3, phenol and p-cresol were not affected by isomalt consumption. In vitro, isomalt was metabolized in several bifidobacteria strains and yielded high butyrate concentrations. Isomalt, which is used widely as a low-glycaemic and low-energy sweetener, has to be considered a prebiotic carbohydrate that might contribute to a healthy luminal environment of the colonic mucosa.
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PMID:Effect of isomalt consumption on faecal microflora and colonic metabolism in healthy volunteers. 1644 15

We present a high-level quantum chemical study of possible elimination reaction mechanisms associated with the catalytic decomposition of urea at the binuclear nickel active site cluster of urease. Stable intermediates and transition state structures have been identified along several possible reaction pathways. The computed results are compared with those reported by Suarez et al. for the hydrolytic catalyzed decomposition. On the basis of these comparative studies, we propose a monodentate coordination of urea in the active site from which both the elimination and hydrolytic pathways can decompose urea into CO2 and NH3. This observation is counter to what has been experimentally suggested based on the exogenous observation of carbamic acid (the reaction product from the hydrolysis pathway). However, this does not address what has occurred at the active site of urease prior to product release. On the basis of our computed results, the observation that urea prefers the elimination channel in aqueous solution and on the observation of Lippard and co-workers of an elimination reaction channel in a urease biomimetic model, we propose that the elimination channel needs to be re-examined as a viable reaction channel in urease.
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PMID:Competitive hydrolytic and elimination mechanisms in the urease catalyzed decomposition of urea. 1767 90

The intensification of grassland systems is leading to serious environmental risks due to the large input of nitrogen (N) in fertilizers and the subsequent gaseous losses. Addition of nitrification inhibitors (NI) or urease activity inhibitors to fertilizers could reduce these losses to the atmosphere. In the present study, the effects of the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) and the urease activity inhibitor N-(n-butyl) thiophosphoric triamide (NBPT) were evaluated on NH3, N2O, NO, and CO2 emissions. Ammonium sulphate nitrate (ASN), urea and cattle slurry were applied at a rate of 70 kg N ha(-1) to a mixed clover-ryegrass sward in the Basque Country (northern Spain) under different soil water contents. NH3 and NO emissions were determined by photoacoustic and chemiluminescence respectively using an open chamber technique while N2O and CO2 emissions were measured by photoacoustic using a closed chamber technique. When the water filled pore space (WFPS) was under 60%, the application of NBPT reduced NO emissions a 34% on urea and an 18% on slurry, and the application of DMPP reduced them a 2% on ASN and a 4% on slurry. No significant effect was observed on NH3 losses. When WFPS was over 60%, no effect could be observed on NO and N2O emissions after the application of both inhibitors, but NH3 losses were reduced a 31% by NBPT when applied with the slurry. Carbon dioxide emissions were unaffected by the use of DMPP or NBPT at any soil water content. Neither grassland yield nor herbage N concentration were influenced by the application of both inhibitors.
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PMID:Effect of N-(n-butyl) thiophosphoric triamide and 3,4 dimethylpyrazole phosphate on gaseous emissions from grasslands under different soil water contents. 1914 92

Manure odor, which results in the increasing complaints and lawsuits, has increased the tension among swine producers and surrounding residents. The effects of Lactobacillus plantarum and different rates of soluble carbohydrates additions to swine manure on odorous compounds, chemical compounds and indigenous flora were evaluated. Additions were calculated on dried manure weight basis. Variables monitored included ammonia (NH3), hydrogen sulfide (H2S), odor offensiveness, pH, ammonium nitrogen (NH4(+)-N), volatile fatty acids (VFAs), urease and indigenous flora. The results indicated that the combination of L. plantarum and soluble carbohydrates dramatically reduced manure pH. Lower pH resulted in the reduction of NH3 volatilization (34.6%-92.4%, P < 0.01), the increases of H2S (P < 0.05) and NH4(+)-N (5.3%-17.5%, P < 0.05). In addition, L. plantarum and soluble carbohydrates additions significantly reduced odor offensiveness, those VFAs related to malodor indicators (valeric acids, 12.3%-47.7%, P < 0.05; iso-valeric, 3.5%-23.8%) and the main microorganisms responsible for odor production, with the number of Eubacteria in swine manure reducing by 4.9%, 11.6%, 17.4%, 34.1% and 32.2% respectively.
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PMID:Effect of adding Lactobacillus plantarum and soluble carbohydrates to swine manure on odorous compounds, chemical composition and indigenous flora. 2005 May 73


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