Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:6.3.4.6 (urease)
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In order to improve the isolation and identification of yeasts in a cancer research hospital, a protocol was developed utilizing an improved blood culture methodology and a four-test schema for rapid yeast identification. The blood culturing technique, based upon centrifugation, has shown a ten-fold increase in isolation of fungi from blood and has provided for: quantitation or organisms, unlimited selection of media and atmospheres for primary culturing, and a 1:200 dilution of microorganisms away from serum antimicrobial factors and antibiotics. The four-test schema, which may be adapted for the identification of any unknown yeast in pure culture, consists of a dye pour plate auxanogram (DPPA), Tween 80-Oxgall-Caffeic acid (TOC), a rapid nitrate-reductase test (swab test) and Urea 'R' Broth. Using this protocol, over 95% of the clinical isolates received were correctly identified within 24 hours and 100% by 48 hours. By using DPPA, a 14 sugar assimilation pattern for each isolate was determined within 12 to 16 hours; and in some cases, as little as 6 hours. Growth on TOC yielded one of the following results: (1) Candida albicans and Candida stellatoidea sequentially produced germ tubes and chlamydospores in 3 hours and 24 hours, respectively; (2) Cryptococcus neoformans produced a brown pigment specific for its identification in 12 hours or less. The swab test gave results on nitrate utilization in less than 15 minutes and urease was detected within 4 hours.
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PMID:Isolation and rapid identification of yeasts from compromised hosts. 37 Jun

The new API 20C yeast identification system together with appropriate microscopic morphology determinations achieved a 97% correlation with a rapid conventional method. Whereas a group composed of Candida, Torulopsis, Saccharomyces, and Rhodotorula was identified with ease (98% overall correlation), a second group, containing Cryptococcus, Trichosporon, and Geotrichum species, appeared to give the system the most difficulty (90% correlation). Within this group particular difficulty was encountered in identifying varieties of Cryptococcus albidus, C. terreus, C. laurentii, Trichosporon beigelli, and Geotrichum spp. as to species. The API 20C system should be incubated the full 72 h prescribed by the manufacturer. However, when used in conjunction with appropriate morphological tests, presumptive identifications of some Candida and Torulopsis species may be made at 24 to 48 h. To facilitate identifications of the more difficult group of yeasts, ancillary tests for determining nitrate reductase, urease, and phenol oxidase activities should be considered as additions to the strip. Incorporating the phenol oxidase test would be especially important for identification of Cryptococcus neoformans, a yeast which should be identified as quickly and as accurately as possible. The API 20C system with computer assistance has proved to be an easy-to-inoculate, versatile, and fairly rapid method of yeast identification, giving results comparable to those obtained by conventional methodologies.
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PMID:Evaluation of the new API 20C strip for yeast identification against a conventional method. 38 21

A rapid simplified screening method that selectively detected the urease activity of 99.6% of 286 isolates of Cryptococcus neoformans within 15 min was developed for use in clinical microbiology laboratories.
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PMID:Rapid selective urease test for presumptive identification of Cryptococcus neoformans. 38 22

We report an urease negative Cryptococcus neoformans derived from pigeon dropping. This isolate produced brown pigmented colonies on cornmeal Tween-80 agar with 300 micrograms/ml caffeic acid, but was failure to hydrolyze urea. More identification tests were performed for this isolate, such as assimilation and fermentation of carbohydrates, nitrate assimilation, production of starch like compound, growth on GCP medium, germ tube formation and inoculation of mice, ect. Most of the results showed that the microbiological characteristics of the isolate were typical of C. neoformans except for negative urease test. Even though there has been a report about an urease negative C. neoformans derived from an AIDS patient, but we have never found any report about isolation from pigeon dropping or nature environment. We should pay attention to the exist of this atypical strain of C. neoformans in nature environment and the possibility of infection to human being. Additionally, we also be aware of the possibility of neglect when this urease negative C. neoformans is identified with urease test.
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PMID:[An urease negative Cryptococcus neoformans]. 141 36

We report a case of fungemia and disseminated disease caused by a urease-negative strain of Cryptococcus neoformans in a patient with the acquired immune deficiency syndrome. Except for failure to hydrolyze urea, the microbiological characteristics of the isolate were typical of C. neoformans. Laboratory specialists should be aware of the occurrence of atypical strains of C. neoformans, particularly those recovered from patients with the acquired immune deficiency syndrome.
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PMID:Disseminated infection caused by urease-negative Cryptococcus neoformans. 305 68

Cryptococcus neoformans var. neoformans (74 isolates) and C. neoformans var. gattii (44 isolates) were used to test urease activity after growth on both yeast extract-glucose-peptone agar (YEPG) and on YEPG supplemented with 100 microM EDTA. Every isolate grown on YEPG agar for 48 h at 30 degrees C produced a positive reaction within 1 h in a modified rapid urease assay at 37 degrees C. However, isolates grown on YEPG with 100 microM EDTA showed a distinct pattern which corresponded to their varietal status. All but 1 of 74 C. neoformans var. neoformans isolates (98.7%) produced a positive reaction within 1 to 4 h, while none of 44 C. neoformans var. gattii isolates produced a positive reaction within the same period. The urease inhibition results and the canavanine-glycine-bromthymol blue agar test results showed 100% correlation among isolates of C. neoformans var. gattii and 98.7% correlation among isolates of C. neoformans var. neoformans. Two representative isolates of C. neoformans var. gattii (serotypes B and C) were further tested for urease during a prolonged incubation period in urea broth. These isolates failed to show a positive reaction even after 11 h of incubation. The uptake of EDTA was negligible in the two varieties. Extracts of cells grown on YEPA agar showed a high level of urease activity in both varieties. Extracts of cells grown on the agar with 100 microM EDTA showed a marked reduction (86%) of urease activity in one isolate of C. neoformans var. gattii but showed only a 30% reduction in one isolate of C. neoformans var. neoformans. Based on these results, the differential effect of EDTA on the two varieties of C. neoformans appeared to be due to greater inhibition of urease synthesis in C. neoformans var. gattii.
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PMID:Urease inhibition by EDTA in the two varieties of Cryptococcus neoformans. 311 9

Ninety-seven strains of Cryptococcus neoformans and C. bacillisporus were examined for 44 biochemical characters and the results were analyzed numerically. One phenon emerged at the 86% level of similarity when strains were clustered according to their M-similarity values. All strains grew in ten carbon sources (D-glucose, D-galactose, arbutin, maltose, sucrose, D-melezitose, D-xylose, D-mannitol, D-glucitol, and meso-inositol), and also grew at 37 degrees C and produced urease and phenoloxidase. None of them grew in melibiose, lactose, nor valine, and none reduced nitrate to nitrite. Comparison of selected biochemical characters, creatinine utilization, and serotypes of 49 aberrant strains is presented. Forty-eight of the 97 strains produced the Filobasidiella state either alone or when paired with a strain of compatible mating-type. Filobasidiella neoformans serotypes A and D were interfertile with compatible mating-types of F. bacillispora serotypes B and C. The 44 biochemical characters and 4 serotypes did not predict barriers to mating competence. The present study further substantiates that Filobasidiella neoformans and F. bacillispora are one species.
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PMID:Biochemical variation of Cryptococcus neoformans. 637 40

Antigen capable of eliciting delayed hypersensitivity reactions in the skin of sensitized guinea pigs could be extracted from Cryptococcus neoformans cells by stirring the cells from 3 to 5 days in concentrated urea or guanidine. Hydrolysis of urea to ammonia by cryptococcal urease accompanied urea extraction, but alkalinity appeared neither necessary nor sufficient for extraction. Antigen from live cells gave larger delayed skin reactions than did antigen from Formalin-killed cells. Peak skin test reactivity appeared to reside in protein-rich fraction having an elution volume on Sephadex G50 corresponding to a molecular weight of 10(4). Activity precipitated with half-saturated ammonium sulfate and could be detected in a single, narrow, rapidly migrating band on disc electrophoresis. Dialyzable proteinaceous antigen and high-molecular-weight, serologically active polysaccharide were present in the antigen, but not active in the delayed hypersensitivity reactions.
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PMID:Cryptococcal skin test antigen: preparation variables and characterization. 701 33

Cryptococcus neoformans strains with various biological characteristics were examinated the phenoloxidase activity on the caffeic acid cornmeal agar (CACA). Other medically important yeasts were also included in this research. Firstly, thirteen reference strains of Cryptococcus neoformans were confirmly found positive phenoloxidase production on the CACA medium. Then, in total 150 isolates of yeasts, forty three Cryptococcus neoformans strains were phenoloxidase positive, other than that 107 other yeast strains were all negative. It is suggested that Cryptococcus neoformans strains with different biological features specifically and uniquely produce phenoloxidase, thus the phenoloxidase test can be applied as a useful tool to identify this pathogenic yeast. Our results also show that the culture on the CACA medium is an effective method to test the phenoloxidase activity within 72 hours. It is further approved that the urease test will not be able to be a screening test for Cryptococcus neoformans in the clinical laboratory, but it is still valuable to identify urease negative Cryptococcus neoformans when the test is used in combination with the phenoloxidase test.
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PMID:[The phenoloxidase test and its application to identify Cryptococcus neoformans with various biological characteristics]. 748 79

This is the first report of a strain of urease negative Cryptococcus neoformans isolated from the environment in China. The colonies of this isolate showed brown pigmentation on cornmeal agar with 300 micrograms/ml caffeic acid, but failed to hydrolyze urea. Microbiological identification and pathogenicity tests in mice confirmed this as a strain of C. neoformans. A similar strain had reportedly been isolated from an AIDS patient.
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PMID:A strain of urease negative Cryptococcus neoformans isolated from the environment in China. 827 25


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