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:6.3.4.6 (urease)
7,490 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

After a brief review of the methods for determination of urea by continuous flow analyzers, a method is described based on the urease splitting of urea followed by NH3 reaction with alfa-ketoglutarate + NADH2 catalysed by GLDH. The method has been applied to continuous flow analyzers and seems to be promising.
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PMID:[Review of the methods of determination of blood urea with continuous-flow analyzers and a proposal of a completely enzymatic UV method for urea by a continuous-flow analyzer]. 718 46

An amount of 0, 0.1, 0.5 and 1.0% (related to N) of phosphoric phenyl ester diamide (PPD) effective as urease blocking substance was applied to the surface of urea with oilbitumen and fed to cows with rumen cannulae over a period of 31 resp. 164 ... 167 days. (The ration essentially consisted of 4.5 kg dried roughage and 1.5 kg starch or 1.1 kg starch plus 0.4 kg sugar and contained 50 g urea). By treating the urea with PPD, the activity of urease, the hydrolysis rate of urea and the NH3-concentration in the rumen were significantly diminished 0.5 to 2 hours after feeding (alpha = 0.05). The effect of PPD was greatest in the first days and decreased with the advancing feeding period. In the variant with 0.5% PPD the examined parameters were significantly reduced after 142 to 164 days, too. This effect remained traceable in its diminished form even after the preparation was discontinued over a period of 35 days. The dynamics of the NH3-concentration was not altered by PPD after a longer feeding period. One can conclude that PPD inhibits the hydrolysis of urea but does not retard it. In conclusion one can say that, because of PPD, the toxicity of urea is lower without the utilisation of urea being better. Due to PPD the molar propionate level in volatile fatty acids decreases significantly and the acetate-propionate relation is expanded.
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PMID:[Studies on the effect of phosphoric phenyl ester diamide as inhibitor of the rumen urease of dairy cows. 1. Influence on urea hydrolysis, ammonia release and fermentation in the rumen]. 728 28

Two lactating dairy cows supplied with rumen and duodenal re-entrance cannulae received with their rations containing 8.2% vegetable crude protein 180 g urea per day (I) or urea treated with the urease inhibitor phosphoric phenyl ester diamide (PPD, 1% of the N-quota). The cows, accustomed to urea, received the PPD-urea without adaptation (II) and after a 30-day adaptation (III) to PPD. On the first day of the experiment one half of the urea was given ruminally in its 15N-labelled form. 2 h after the isotope supplementation 62.5 and 24 mg NH3 and 6.58 and 21 mg urea/100 ml could be detected in the rumen juice of I to III. Within 72 h 16.6, 26.1 and 25.2% of the 15N-excess given (15N') passed the duodenum in TCA - soluble form and 31.2, 28.4 and 41.7% in TCA - precipitable form. 15.6, 24.4 and 21.5% of the total amount of 15N were excreted in urine and 4.5, 4.6 and 6.0% in the milk protein. The values for faeces were 14.4, 14,4 and 15.4%. The conclusion from these results and from the dynamics of the relative 15N' in the fractions of the rumen fluid is that with a limited inhibition of rumen urease by PPD as it develops after the adaptation, the utilisation of urea-N can be improved.
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PMID:[Studies on the effect of phosphoric phenyl ester diamide as inhibitor of the rumen urease of dairy cows. 2. The metabolism of 15N-urea]. 728 29

The factors that determine which Helicobacter pylori infected subjects develop duodenal ulcer (DU) are unclear. This study tested the hypothesis that infection density and urease activity are higher in DU than non-DU subjects. Fifty five DU and 55 age and sex matched non-DU subjects were studied. Quantitative methods were used for measuring infection density (viable organism count) and urease activity (Berthelot reaction). DU subjects had a greater antral infection density (geometric mean of colony forming units/mg biopsy protein; 10.5 x 10(5) v 1.3 x 10(5), p < 0.001). They also had higher biopsy urease activity (geometric mean of NH3 nmol/min-1/mg protein-1; 103 v 25, p < 0.001). Urease activity per organism, however, was similar in the two groups showing that high antral urease activity in DU was a reflection of organism density. DU was not present in subjects with an antral infection density less than 10(5) colony forming units/mg protein. A correlation was present between H pylori viable counts and the severity and activity of gastritis. Both severity and activity of gastritis were greater in the antrum of DU compared with non-DU subjects but there was no difference in the body between the two groups. It is concluded that antral H pylori infection density is probably an important determinant of DU development, and that there is a baseline of infection density that is necessary for ulcer formation.
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PMID:Helicobacter pylori infection density and gastric inflammation in duodenal ulcer and non-ulcer subjects. 870 3

Urease is a virulence determinant, a taxonomic and diagnostic marker, and immunogen for Helicobacter pylori, an aetiologic agent of gastritis and peptic ulceration. This enzyme requires Ni2+ ions in the active site for successful hydrolysis of urea. When expressed in Escherichia coli, recombinant urease is only weakly active unless urease structural subunits are overexpressed, exogenous NiCl2 is added, and the host strain is grown in medium that does not chelate free Ni2+. As wild-type H. pylori does not require such conditions for very high levels of urease expression, we reasoned that additional genes were required to accumulate the metal ion. To isolate such genes, E. coli SE5000 (pHP808), which carries the H. pylori urease gene cluster, was complemented with a lambda ZAP-derived plasmid library of the H. pylori chromosome. One of 1000 ampicillin-resistant clones, plated onto urea segregation agar, produced detectable urease. Urease activity of this co-transformant, grown in Luria broth containing 1 microM NiCl2, was 36 mumol NH3 min-1 mg-1 protein. Urease-enhancing activity, which is not directly linked to the urease gene cluster, was localized by subcloning and nucleotide sequencing. The largest open reading frame, designated nixA, predicted a polypeptide of 34,317 Da that displayed characteristics of an integral membrane protein. In vitro transcription-translation of nixA sequences yielded a polypeptide estimated to be 32 kDa in size. An in-frame Bal31 deletion within nixA abolished urease-enhancing activity. At 50 nM NiCl2, E. coli containing the nixA clone transported 1250 +/- 460 pmol Ni2+ min-1 10(-8) cells, whereas the vector control transported only 140 +/- 85 pmol Ni2+ min-1 10(8) cells, i.e. significantly less (P = 0.01). We conclude that NixA confers upon E. coli a high-affinity nickel-transport system (KT = 11.3 +/- 2.4 nM; Vmax = 1750 +/- 220 pmol Ni2+ min-1 10(-8) cells) and is necessary for expression of catalytically active urease, regardless of growth conditions.
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PMID:Helicobacter pylori nickel-transport gene nixA: synthesis of catalytically active urease in Escherichia coli independent of growth conditions. 765 Nov 42

Helicobacter pylori was transurethrally inoculated into the mouse urinary tract. The organism established infection and induced inflammation in the urinary bladder and pelvis. During the infection, urinary pH was elevated, probably due to the production of NH3 by bacterial urease. H. pylori was recovered from the urinary bladder, kidney and urine of the infected mice. Histopathologically, severe neutrophil infiltration was observed in the mucosal layer of both organs. H. pylori was detected on the surface of the epithelial cells. These results indicate that low pH and bacterial flora were not essential factors in establishing the mucosal infection with H. pylori. This experimental system is useful to investigate the pathogenicity of H. pylori in mucosal organs.
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PMID:Helicobacter pylori induces inflammation in mouse urinary bladder and pelvis. 793 56

A low ventilation model to induce ascites was introduced and characterized. In addition, the effect of supplemental air mixing via ceiling fans (CF) and the feeding of a urease inhibitor (0, 125, and 250 ppm) on incidence of ascites were investigated. Twelve environmental chambers were utilized in the trial; six were fitted with CF. Each dietary treatment was replicated twice per CF treatment. One hundred and twenty day-old male commercial broilers were reared per chamber. Atmospheric O2, CO2, and NH3, temperature, and humidity, as well as weekly litter moisture and pH, were monitored. Chamber CO2 levels increased immediately then stabilized. Chamber NH3 levels increased between 2 to 4 wk of age and rapidly declined when ventilation rates were increased to 1 cfm per bird. The CF and dietary treatments had little effect on air or litter variables except for NH3. Supplementing the diet with urease inhibitor resulted in a greater than 50% reduction in cumulative mortality due to ascites and a slight reduction in weekly BW gains. The CF treatment had no effect on production variables such as weekly feed intake, gain, and feed to gain ratio, or survivability due to ascites.
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PMID:Effect of a urease inhibitor and ceiling fans on ascites in broilers. 1. Environmental variability and incidence of ascites. 807 22

Texel wethers (68 +/- 2.5 kg BW) fitted with catheters in the ruminal veins and a mesenteric artery, blood flow probes on ruminal arteries, and a ruminal cannula were fed 500 g of orchardgrass hay every 12 h. During the last third of the feeding cycle, intraruminal injections were performed to evaluate the effect of urease activity, osmolality, and concentrations of NH3, butyrate, and CO2 in the rumen on urea and NH3 fluxes across the rumen wall. At pH 6.7, NH3 absorption increased with NH3 and butyrate concentrations in the rumen, and to a lesser extent with CO2 concentration. The increase in ruminal blood flow associated with CO2 and butyrate increase was always greater than the increase in NH3 absorption. Increasing ruminal osmolality slightly decreased NH3 absorption. Ruminal NH3 concentration and ruminal blood flow seemed to be the main determinant of NH3 absorption. Decreasing urease activity in the rumen decreased urea net transfer. The net transfer of urea to the rumen was stimulated by CO2. High concentrations of NH3 (330 mg of N/L) and butyrate (25 mM) in the rumen decreased urea net uptake, whereas osmolality (up to 420 mOsmol/L) did not affect it. Modifications in ruminal blood flow or water net movement across the ruminal wall did not seem to account for the effect of CO2, NH3, and butyrate on urea net uptake.
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PMID:Net transfer of urea and ammonia across the ruminal wall of sheep. 822 81

The in vitro effect of urea and hydrolysis of urea by urease on mucus H+ permeability is reported here. The effective DHCl values indicate a strong pH dependence for H+ diffusion in both water and mucus layers, with no apparent trend at concentrations between 1 and 50 mM urea. However, the estimated DHCl at near-neutral and alkaline pH are 4- to 10-fold lower through mucus than through aqueous films. Moreover, the pKa values of HCO3- and NH3 (generated by urease action on urea) had a profound effect on measured DHCl. These in vitro studies suggest that a high local concentration of NH3 and HCO3- within the mucus layer, generated by the action of Helicobacter pylori urease on endogenous intragastric urea, could greatly accelerate proton flux to the surface epithelium by operation of a buffer shuttle. This results in enhanced H+ permeability, particularly at pKa values of HCO3- and NH3, and in extreme circumstances it may result in gastric ulcer formation.
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PMID:An in vitro study of enhanced H+ diffusion by urease action on urea. Implications for Helicobacter pylori-associated peptic ulceration. 826 22

In the rat kidney, arginine (Arg) synthesis is restricted to the proximal tubule with a decreasing intensity from its convoluted (PCT) to its straight part (PST). The present study was designed to investigate the pattern of Arg synthesis along the nephron in other mammals, the mouse and rabbit. Microdissected representative nephron segments were incubated with 0.1 mM L-[ureido-14C]citrulline in a sealed chamber. Addition of arginase and urease to the incubation medium led to the hydrolysis of Arg into ornithine, NH3, and 14CO2. The latter was trapped in KOH and counted (results are in fmol Arg.min-1.mm tubular length-1). As in the rat, the main site of Arg synthesis in both species was found to be the PCT (mouse, 191; and rabbit, 57). A lower production was observed in rabbit and mouse PST and in rabbit distal segments. Along the PCT (from 1st to 4th mm after the glomerulus), a steep decrease is observed in mouse (595 and 37, respectively) but not in rabbit (57 and 23). The fate of the newly synthesized Arg probably depends on its site of production. Intracellular arginase activity is known to be present in the cortical (C) and medullary (OS) PST, in both mouse and rabbit. In rabbit only, arginase activity is also found in the PCT. We observed that a large part of Arg was further hydrolyzed into urea and ornithine in CPST and OSPST of mouse (66 and 80%, respectively) and rabbit (40 and 70%) but not in rabbit PCT (8%). Thus Arg produced by PCT in both species is probably released in the cortical blood, whereas Arg produced in PST may serve locally to produce urea and ornithine, and the latter could be used for polyamine synthesis.
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PMID:Arginine synthesis in mouse and rabbit nephron: localization and functional significance. 832 90


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