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)

A total of 16 strains of Enterohemorrhagic Escherichia coli (EHEC) isolated from diarrea patients in Saitama from 1990 to 1992 were tested for their serotype, verotoxin production, biochemical characteristics, antibiotics sensitivity and plasmid profiles. By serotype analysis, 14 strains from two outbreaks and 12 sporadic cases were classified as type O157:H7, one as O111:H-(not motility) and one as O128:H2. Typing of verotoxin by gene analysis using Polymerase Chain Reaction (PCR) showed that 9 of O157:H7 strains including two cases from outbreaks and O128:H2 have VT1 and VT2 genes, other O157:H7 have the VT2 gene and O111:H-has only the VT1 gene. Biochemical characteristic analysis indicated two strains of O157:H7 type from outbreaks were biotype II and the rest of O157:H7 were biotype I. One of the O157:H7 strain from a sporadic case showed positive for urease production. According to sensitivity tests against antibiotics, out of the O157:H7 group, one strain was resistant against ABPC, one against SM and two strains resistant to SM-TC. For plasmid profiles, all strains had 94 Kb plasmids and several smaller sizes of plasmids. But 5 strains of O157:H7 had 94 Kb plasmid only.
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PMID:[Biological characters of enterohemorrhagic Escherichia coli isolates from diarrhea patients in Saitama (1990-1992)]. 787 66

Polymerase chain reaction (PCR) amplification of the urease genes of Ureaplasma urealyticum was compared with culture for detection of the organism in 100 endotracheal aspirates from 54 ventilated preterm infants. Ninety specimens gave negative results by both culture and PCR and three specimens gave positive results by both culture and PCR. Six specimens were negative by culture but positive by PCR. The one specimen positive by culture and negative by PCR was interpreted as a false-positive culture result. Overall agreement between results obtained by culture and PCR was 93%. PCR is a sensitive and reliable method for the detection of U. urealyticum in neonatal endotracheal secretions. Detection by PCR (1-2 days) is more rapid than culture (2-5 days) and this will be important if early therapeutic intervention is shown to be effective.
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PMID:Comparison of culture with the polymerase chain reaction for detection of Ureaplasma urealyticum in endotracheal aspirates of preterm infants. 866 7

CURRENT DIAGNOSTIC METHODS: Helicobacter pylori infection plays a central role in the pathophysiology of gastrointestinal disease, and its accurate diagnosis and successful eradication is crucial in a wide range of different circumstances. Currently, serology is recommended for initial screening, followed by histology and/or culture to confirm the diagnosis before treatment. Since H. pylori is developing greater resistance to certain antibiotics, culture is becoming increasingly important in some populations to test for susceptibility to antibiotics. To confirm eradication after treatment, the urea breath test is used. This test is presently the best non-invasive test to determine eradication. NEW APPROACHES: Considerable efforts are being made to improve diagnostic methods, and a host of new or improved approaches can be expected in the near future. For general screening, tests are being developed that use whole blood and can be used by general practitioners to give rapid results in a cost-effective manner. The evidence so far suggests that these new 'office' tests are not as accurate as laboratory tests, but they are nevertheless important for general diagnostic purposes. Serological tests for cagA antibodies and immunoblot tests are also under development. New biopsy-based tests include the development of a true rapid urease test which will give accurate results in 1 h. Polymerase chain reaction/DNA enzyme immunoassay detection is another field receiving attention. Non-invasive direct tests of the future are likely to include the use of the polymerase chain reaction in faeces. This paper reviews current diagnostic modalities for H. pylori and gives an overview of expected future developments.
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PMID:The most important diagnostic modalities for Helicobacter pylori, now and in the future. 2249 1

We report, for the first time, the presence in Helicobacter pylori of an aliphatic amidase that, like urease, contributes to ammonia production. Aliphatic amidases are cytoplasmic acylamide amidohydrolases (EC 3.5.1.4) hydrolysing short-chain aliphatic amides to produce ammonia and the corresponding organic acid. The finding of an aliphatic amidase in H. pylori was unexpected as this enzyme has only previously been described in bacteria of environmental (soil or water) origin. The H. pylori amidase gene amiE (1017 bp) was sequenced, and the deduced amino acid sequence of AmiE (37746Da) is very similar (75% identity) to the other two sequenced aliphatic amidases, one from Pseudomonas aeruginosa and one from Rhodococcus sp. R312. Amidase activity was measured as the release of ammonia by sonicated crude extracts from H. pylori strains and from recombinant Escherichia coli strains overproducing the H. pylori amidase. The substrate specificity was analysed with crude extracts from H. pylori cells grown in vitro; the best substrates were propionamide, acrylamide and acetamide. Polymerase chain reaction (PCR) amplification of an internal amiE sequence was obtained with each of 45 different H. pylori clinical isolates, suggesting that amidase is common to all H. pylori strains. A H. pylori mutant (N6-836) carrying an interrupted amiE gene was constructed by allelic exchange. No amidase activity could be detected in N6-836. In a N6-urease negative mutant, amidase activity was two- to threefold higher than in the parental strain N6. Crude extracts of strain N6 slowly hydrolysed formamide. This activity was affected in neither the amidase negative strain (N6-836) nor a double mutant strain deficient in both amidase and urease activities, suggesting the presence of an independent discrete formamidase in H. pylori. The existence of an aliphatic amidase, a correlation between the urease and amidase activities and the possible presence of a formamidase indicates that H. pylori has a large range of possibilities for intracellular ammonia production.
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PMID:Identification and characterization of an aliphatic amidase in Helicobacter pylori. 936 23

NixA, the high affinity nickel transport protein of Helicobacter pylori, imports Ni2+ ions across the cytoplasmic membrane for insertion into the active site of the urease metalloenzyme, which is essential for colonization of the gastric mucosa. Twelve conserved aspartate (aspartates 47, 49, 55, 194, 231, and 234), glutamate (glutamates 106, 198, and 274), and histidine (histidines 44, 50, and 79) residues were identified by alignment of NixA with homologous transporters. Polymerase chain reaction-generated site-directed mutants of these residues were expressed in E. coli along with the H. pylori urease gene cluster. Mutations in residues within the predicted periplasmic domains of NixA maintained near wild type levels of Ni2+ uptake and urease activity, as did control mutations of conserved positively charged residues (lysines 140 and 268; arginines 162 and 167). Mutations in highly conserved motifs in predicted helices II and III of NixA abolished Ni2+ uptake and urease activity. Mutations in helices V and VI and the cytoplasmic domains decreased Ni2+ transport rates by >/=90%. Reduction in rates of Ni2+ transport correlated with reduction in urease activities (r = 0.77). Ni2+ transport was inhibited in the presence of Co2+, Cu2+, and Zn2+, indicating that these ions may also be bound or transported by NixA. We conclude that conserved Asp, Glu, and His residues in the transmembrane domains of NixA are critical for the transport of the divalent cations Ni2+, Co2+, Cu2+, and Zn2+ into the cytoplasm of H. pylori.
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PMID:Conserved residues and motifs in the NixA protein of Helicobacter pylori are critical for the high affinity transport of nickel ions. 941 70

The aim of this study was to identify the natural reservoir and route of transmission of Helicobacter pylori infection. Two hundred eight (208) dyspeptic patients (114 males, 94 females; peak age of cohort, 50-59.9) were recruited. Specimens were collected from saliva, supra- and subgingival dental plaque, tongue scrapings, and oropharyngeal swabs. At subsequent endoscopy, gastric antral biopsy was performed for the rapid urease test (RUT), microbiological culture, and, in some patients, histology. Gastric juice samples were aspirated, and in 50 patients duodenal aspirate was collected. Polymerase chain reaction (PCR) with primers targeted to the 16S rRNA sequence of H. pylori was also employed for each of the specimens. In those patients where H. pylori was detected from multiple sites (dental plaque, gastric juice, gastric biopsy, and duodenal aspirate), restriction endonuclease digestion with Hae III was performed to determine if they were epidemiologically linked. The results indicated that 15/208 patients (7%) tested positively for H. pylori by PCR in dental plaque; only 2 samples were positive by culture. In none of the other oral sites sampled was H. pylori detected by any test used in the study. Gastric juice and gastric biopsy specimens from 36/ 208 patients (17%) and 114/208 patients (55%), respectively, were positive by PCR. Duodenal aspirate from 6/50 patients (12%) also tested positively by PCR. All specimens tested by restriction endonuclease digestion with Hae III (15/15 patients) were positive in both antral biopsy and gastric juice specimens, as well as 5 specimens from the duodenal aspirate. Four of the dental plaque strains had restriction patterns similar to those of the stomach and duodenal sites, providing evidence that these sites were infected with the same strain of H. pylori. In conclusion, the results suggest that H. pylori selects the gastric mucosa as its preferred site. The detection in dental plaque could indicate that the oral cavity may act as a reservoir or sanctuary for the organism. Whether H. pylori is a resident or transient oral microorganism is still unclear, although it is more likely to be transient in nature.
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PMID:Helicobacter pylori: the mouth, stomach, and gut axis. 972 11

Several species of Helicobacter colonize the hepatobiliary tract of animals and cause hepatobiliary diseases. The aim of this study is to investigate Helicobacter found in the biliary tract diseases of humans. Thirty-two bile samples (15 from bile duct cancer, 6 from pancreatic head cancer, and 11 from intrahepatic duct stone) were obtained by percutaneous transhepatic biliary drainage. Polymerase chain reaction analysis using Helicobacter specific urease A gene and 16S rRNA primers, bile pH measurement, and Helicobacter culture were performed. Helicobacter DNA was detected in 37.5%, and 31.3% by PCR with ureA gene, and 16S rRNA, respectively. The bile pH was not related to the presence of Helicobacter. The cultures were not successful. In conclusion, Helicobacter can be detected in the bile of patients with bile duct diseases. The possibility of pathogenesis of biliary tract diseases in humans by these organisms will be further investigated.
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PMID:Detection of Helicobacter DNA in bile from bile duct diseases. 1033 65

In this study, we evaluated the usefulness of a culture system used in our laboratory for the detection of Ureaplasma urealyticum in the endotracheal aspirates of neonates. Ureaplasmal broth was used to enhance the growth of U. urealyticum followed by observation of colonies on A7 agar. Of the 68 samples of endotracheal aspirates tested, 60 gave positive indication of urease activity by the broth. However, only 14 yielded U. urealyticum colonies on A7 agar medium. Polymerase chain reaction detected U. urealyticum in 21 samples. The use of Ureaplasmal broth was therefore not specific for the diagnosis of U. urealyticum. We suggest that subculture onto agar medium or PCR is essential for definite identification of U. urealyticum.
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PMID:An evaluation of the usefulness of a culture system for the detection of Ureaplasma urealyticum in the endotracheal aspirates of neonates. 1087 44

Polymerase chain reaction assay using ureC gene specific primers for the detection of Helicobacter pylori in gastric biopsy specimens from 116 dyspeptic patients was compared with other routine invasive diagnostic methods (culture, rapid urease test [RUT] and histology). In parallel, gastric biospy specimens from 54 patients and their corresponding Helicobacter pylori isolates were subjected to PCR with cagA targeting primers using standard protocols. Helicobacter pylori were detected in 53%, 43%, 48% and 50% of patients by PCR, RUT, culture and histological examination respectively. Based on histology and culture positive and at least three test positive result, 44 (37%), 46 (39%) and 26 (22%), and 56 (48%), 52 (44%) and 8 (6%) patients were classified as Helicobacter pylori positive, negative and indeterminate respectively. The sensitivity and specificity of PCR assay was the highest-95% and 100% when compared with both culture and histology positive, and at least any three positive results respectively. The result of cagA positivity in 54 gastric biopsy specimens and their corresponding Helicobacter pylori isolates were identical; 18 of 20 (90%) duodenal ulcer patients and 23 of 28 (82%) patients with chronic gastritis and 2 (40%) of 5 patients with portal hypertension and one gastric biopsy specimens from gastric cancer patients were found to be cagA positive. PCR-based method to detect Helicobacter pylori and the virulence gene cag A directly from gastric biopsy specimens appears to be promising and can curtail the lengthy process of culture-based approaches. The procedure proved to be rapid and reliable and could be utilized for diagnostic purposes.
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PMID:UreC PCR based diagnosis of Helicobacter pylori infection and detection of cag A gene in gastric biopsies. 1259 61

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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