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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)

Four Helicobacter pylori strains were used to develop in vitro methods to assess adherence to HeLa cells. Using direct detection by microscopy, adhesion scores increased with the initial bacteria-to-cell ratio. The urease method assessed H. pylori bound to HeLa cells by their urease activity. The percentage of the original inoculum adhering to HeLa cells remained constant for initial ratios from 10(2) to 10(5) bacteria per cell. An ELISA using anti-H. pylori serum assessed whole bacteria or components bound to HeLa cell fractions. By all three methods, the four H. pylori strains were adherent to HeLa cells or membranes whereas Campylobacter fetus and Providencia control strains were not. The adherence of H. pylori whole cells decreased following extraction with saline, water, or glycine buffer and most of the superficial adhering material (SAM) was present in the saline or water extracts. SAM bound better to HeLa membranes than to calf fetuin or bovine serum albumin (BSA); binding was inhibited by preincubation of SAM with HeLa membranes but not with fetuin or BSA or by pretreatment of HeLa membranes with neuraminidase. These data indicate that SAM has a specific receptor on the HeLa cell membranes. By gel exclusion chromatography of bacterial extracts, the most adherent components were found in the fractions which also contained the highest urease activity; these fractions included urease subunit antigens. We conclude that adherence of H. pylori can be assessed by microtiter assays and involves bacterial surface material which co-purifies with urease and is different from the N-acetyl-neuraminyl-lactose binding hemagglutinin.
Microb Pathog 1990 Dec
PMID:Adherence of Helicobacter pylori cells and their surface components to HeLa cell membranes. 209 96

Helicobacter pylori (formerly Campylobacter pylori) has been recently described as a gastritis-associated bacterium. We examined endoscopic biopsies of 100 patients with dyspepsia and found H. pylori in the gastric antrum of 34 (34%) by either culture, urease tests and/or histology. Thirty-one out of 41 patients (75.6%) confirmed to have chronic active gastritis histologically had H. pylori in their gastric antrum compared to 3 out of 59 patients (5.1%) with dyspepsia but normal histology (p less than 0.01). Histological examination, using gram stain and the Warthin-Starry Silver stain, detected 29 of the 34 positive cases (85.3%); urease test, 26 cases (76.5%) and culture, 22 cases (64.7%). A combination of histological examination and urease test increased the detection rate to 97.1%. Therefore we felt that for the detection of H. pylori in endoscopic biopsies, culture, which is time consuming and expensive, is not necessary in routine diagnosis as it did not improve the diagnostic rate over a combination of histology and urease test. A comparative study on three media (blood agar, chocolate agar and Skirrow's agar) used in the isolation of the organism showed that non-selective blood agar and chocolate agar were superior to Skirrow's agar. The strains isolated appeared to be homogeneous in their morphological and biochemical characteristics.
Malays J Pathol 1990 Dec
PMID:Detection of Helicobacter pylori from endoscopic biopsies and the biochemical characteristics of these isolates. 210 69

Studies with two uropathogenic urease-producing Escherichia coli strains, 1021 and 1440, indicated that the urease genes of each are distinct. Recombinant plasmids encoding urease activity from E. coli 1021 and 1440 differed in their restriction endonuclease cleavage sites and showed minimal DNA hybridization under stringent conditions. The polypeptides encoded by the DNA fragments containing the 1021 and 1440 urease loci differed in electrophoretic mobility under reducing conditions. Regulation of urease gene expression differed in the two ureolytic E. coli. The E. coli 1021 locus is probably chromosomally encoded and has DNA homology to Klebsiella, Citrobacter, Enterobacter, and Serratia species and to about one-half of the urease-producing E. coli tested. The E. coli 1440 locus is plasmid encoded; plasmids with DNA homology to the 1440 locus probe were found in urease-producing Salmonella spp., Providencia stuartii, and two E. coli isolates. In addition, the 1440 urease probe was homologous to Proteus mirabilis DNA.
J Bacteriol 1990 Dec
PMID:Genetic analysis of Escherichia coli urease genes: evidence for two distinct loci. 217 68

In the filtrate and/or dialysate regeneration system, which is expected to miniaturize the artificial kidney, ammonium ion decomposed from urea by immobilized urease is removed competitively by ion-exchangers from coexisting cations. Since divalent cations such as Ca2+ and Mg2+ are more favorably exchanged than ammonium ion, this system needs supplementation of these cations and, thus, additional amount of ion-exchanger. To minimize these requirements, we utilized positively charged membrane to process cation-free filtrate in which urea is dialyzed. Positively charged membranes were tested in vitro to evaluate separation efficiency between urea and cations. Equilibrium adsorption of ammonium ion to ion-exchanger with or without co-existing cations demonstrates that this filtrate regeneration system can reduce the amount of ion-exchanger up to one-half to one-third of that of the conventional system. In an ex vivo experiment with a mongrel dog, blood urea nitrogen (BUN) concentration was maintained at the same level or up to 23% less than the initial value.
Artif Organs 1990 Dec
PMID:A new approach for the filtrate regeneration system in the wearable artificial kidney. 217 68

We have constructed an opal suppressor system in Escherichia coli to complement an existing amber suppressor system to study the structural basis of tRNA acceptor identity, particularly the role of middle anticodon nucleotide at position 35. The opal suppressor tRNA contains a UCA anticodon and the mRNA of the suppressed protein (which is easily purified and sequenced) contains a UGA nonsense triplet. Opal suppressor tRNAs of two tRNA(Arg) isoacceptor sequences each gave arginine in the suppressed protein, while the corresponding amber suppressors with U35 in their CUA anticodons each gave arginine plus a second amino acid in the suppressed protein. Since C35 but not U35 is present in the anticodon of wild-type tRNA(Arg) molecules, while the first anticodon position contains either C34 or U34, these results establish that C35 contributes to tRNA(Arg) acceptor identity. Initial characterizations of opal suppressor tRNA(Arg) mutants by suppression efficiency measurements suggest that the fourth nucleotide from the 3' end of tRNA(Arg) (A73 or G73 in different isoacceptors) also contributes to tRNA(Arg) acceptor identity. Wild-type and mutant versions of opal and amber tRNA(Lys) suppressors were examined, revealing that U35 and A73 are important determinants of tRNA(Lys) acceptor identity. Several possibilities are discussed for the general significance of having tRNA acceptor identity in the same positions in different tRNA acceptor types, as exemplified by positions 35 and 73 in tRNA(Arg) and tRNA(Lys).
Proc Natl Acad Sci U S A 1990 Dec
PMID:Nucleotides that determine Escherichia coli tRNA(Arg) and tRNA(Lys) acceptor identities revealed by analyses of mutant opal and amber suppressor tRNAs. 225 Dec 70

We evaluated the diagnostic accuracy of endoscopic finding of nodular antritis and rapid urease test (RUT) in order to simplify the approach to the diagnosis of Helicobacter pylori (H. pylori) infection. Forty-four consecutive patients (mean age 7.9 yr, range 6-13 yr) referred because of recurrent abdominal pain as the main symptom, were prospectively investigated for the presence of H. pylori. H. pylori positivity or negativity was defined as the concordance of two of the following tests: RUT, microbiologic culture, and histologic examination on bioptic samples. RUT sensitivity was 100%, whereas specificity was 87.5%. The presence of nodular antritis had a sensitivity of 96.4% and specificity of 87.5% in H. pylori infection diagnosis. The predictivity value of combined RUT and nodular antritis, whether positive or negative, was 100%. Only in case of discordance do we suggest the utilization of other expensive tools for diagnosis of H. pylori infection.
Am J Gastroenterol 1990 Dec
PMID:Helicobacter pylori infection: a simplified diagnostic approach. 188 14

Arginine production was measured in isolated rat nephron segments. Segments were incubated with 0.3 mM aspartate and 0.1 mM L-[ureido-14C]-citrulline in a sealed chamber. Arginase and urease were added to the medium to hydrolyze arginine and to release 14CO2, which was trapped in KOH and counted. Arginine synthesis was found only in the proximal tubule, with decreasing intensity from proximal convoluted (PCT) to proximal straight tubule (PST). Results were as follows (in fmol.min-1.mm tubule length-1): PCT, 122 +/- 15; cortical PST, 71 +/- 6; outer medullary PST, 41 +/- 4; all other segments, less than 6. Arginine synthesis changed almost proportionally with precursor concentration of less than or equal to 0.4 mM. We had shown previously that PST but not PCT was able to hydrolyze arginine into urea and ornithine. In this study arginine was further hydrolyzed in cortical (40%) and medullary (64%) PST but not in PCT. These observations suggest that the arginine formed in PCT contributes to the maintenance of the whole body arginine pool, whereas most of the arginine formed in PST might contribute, by its conversion to urea, to the process of urine concentration.
Am J Physiol 1990 Dec
PMID:Localization of arginine synthesis along rat nephron. 226 Jun 85

CDC group M-6 is the vernacular name given to a gram-negative, oxidase-positive, aerobic, nonmotile, rod-shaped bacterium. This organism is biochemically similar to Kingella denitrificans and displays a cellular fatty acid profile consistent with CDC groups M-5 and EF-4 and with Neisseria elongata. Of the 95 M-6 strains referred to the Centers for Disease Control (CDC) for identification, 32 (64%) of the first 50 were from the throat or sputum and only 3 (6%) were from blood; only 5 (11%) of the next 45 isolates were from the upper respiratory tract and 23 (51%) were from blood, with many of these (15 or 65%) being associated with endocarditis. The major characteristics of CDC group M-6 include reduction of nitrate and nitrite with no gas formation; positive reaction for oxidase; negative reactions for catalase, urease, indole, and motility; and no acid production from carbohydrates. Guanine-plus-cytosine content determined spectrophotometrically by thermal denaturation was 55 to 58 mol % for six M-6 strains tested: 56 mol % for the N. elongata subsp. elongata type strain and for the N. elongata subsp. glycolytica type strain. By the hydroxyapatite method, DNAs from 24 M-6 strains showed an average of 78% relatedness to M-6 reference strain B1019 in reactions at 60 degrees C and 73% relatedness in reactions at 75 degrees C. M-6 strain B1019 was 79% related to the N. elongata type strain at 60 degrees C and 71% related at 75 degrees C; it was 75% related to the type strain N. elongata subsp. glycolytica at 60 degrees C and was 66% related at 75 degrees C. DNAs from CDC group EF-4, K. denitrificans, and CDC group M-5 were all less than 14% related to CDC group M-6 at 75 degrees C. The DNA relatedness data showed conclusively that all the M-6 strains belong in the species N. elongata. M-6 is different from N. elongata subsp. elongata in that M-6 reduces nitrate and sometimes weakly acidifies D-glucose, and it is different from N. elongata subsp. glycolytica in that it reduces nitrate and is negative for glucose and catalase. Because of the apparent clinical significance of M-6 compared with the clinical significance of N. elongata subsp. elongata and N. elongata subsp. glycolytica and the ease in distinguishing it biochemically, we propose M-6 as a third subspecies of N.elongata, N. elongata subsp. nitroreducens subsp. nov.
J Clin Microbiol 1990 Dec
PMID:Neisseria elongata subsp. nitroreducens subsp. nov., formerly CDC group M-6, a gram-negative bacterium associated with endocarditis. 227 87

Sera from 100 children (ages, 6 to 16 years) presenting with upper gastrointestinal symptoms were examined for antibodies to Helicobacter pylori by enzyme-linked immunosorbent assay (ELISA) based on crude, loosely cell-associated antigens and a partially purified urease antigen preparation. All children underwent endoscopy, and 20 children were shown to have H. pylori infection by histology or direct culture. Serum anti-H. pylori immunoglobulin G (IgG) levels (crude antigen) were clearly raised in the infected group, particularly after preabsorption of sera against a Campylobacter jejuni antigen preparation, while IgM and IgA ELISA determinations did not discriminate between infected and H. pylori-negative patients. Only 14 children in the infected group had raised anti-urease IgG levels. Two patients in whom the organism was not demonstrated or cultured had raised specific IgG levels against both crude and urease antigens and pathological features consistent with H. pylori disease. Immunoblotting studies did not reveal any single protein antigen or simple combination of antigens that could be considered as a candidate for a more defined serodiagnostic reagent. Anti-H. pylori antibody determinations (crude antigen) performed on posttreatment samples from children in whom the organism could no longer be demonstrated suggested that sustained IgG levels may not be a reliable index of treatment failure. An IgG ELISA based on crude, loosely cell-associated antigens of H. pylori can be used for the serodiagnosis of H. pylori infection in childhood.
J Clin Microbiol 1990 Dec
PMID:Serodiagnosis of Helicobacter pylori infection in childhood. 227 95

We have developed a novel and practical DNA-RNA hybridization assay for the detection and identification of Campylobacter pylori in the gastric mucosa. This technique utilizes a [32P]ddATP-labeled synthetic oligonucleotide probe complementary to a nucleotide sequence present in C. pylori 16S rRNA. This probe is very sensitive and reacted with all 23 strains of C. pylori tested. It is also highly specific, since there was no cross-reactivity with the heterologous organisms Campylobacter coli, C. fetus subsp. fetus, C. jejuni, and C. laridis or with Escherichia coli. Hybridization of the oligonucleotide probe with C. pylori RNA was completely inhibited by treatment of the membrane filters with RNase but not DNase. Although a gastric mucosa tissue homogenate slightly inhibited the hybridization, as few as 10(4) C. pylori cells could be detected even in the presence of 5 mg of gastric mucosa. Gastric biopsy specimens obtained from patients referred for upper gastrointestinal tract endoscopy were tested for C. pylori infection by direct oligonucleotide hybridization, and the results were compared with those of bacteriological cultures, the urease test, and histological observations. A comparison of the urease test and the oligonucleotide hybridization results showed an excellent correlation between the two methods. The clinical usefulness of this oligonucleotide-RNA hybridization method is discussed.
J Clin Microbiol 1989 Dec
PMID:Oligonucleotide probe for detection and identification of Campylobacter pylori. 248 Mar 60


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