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An assay was developed for the specific detection of Salmonella enterica serotype Enteritidis, using a novel application of the polymerase chain reaction (PCR). This PCR assay is based on the mismatch amplification mutation assay, an allele-specific reaction, and can discriminate Enteritidis from all other salmonella. PCR primers were selected to amplify a 351-base pair (bp) DNA fragment from the salmonella plasmid virulence A (spv A) gene of Enteritidis. A single base difference at position 272 is present between the nucleotide sequence of the spvA gene of Enteritidis and other salmonellae. The downstream PCR primer, that encompasses position 272 of the Enteritidis spvA gene, was designed to contain a single base mismatch at the penultimate position, resulting in a 1-base mismatch with Enteritidis and a 2-base mismatch with other salmonellae that harbour the virulence plasmid. The upstream primer was completely homologous with the region immediately 5' to the spvA gene. When these primers were used and the annealing and extension reactions were performed at the same temperature, the PCR assay was specific for Enteritidis; no PCR product was detected for 40 other serotypes and 28 different genera examined. In pure culture, 120 colony forming units (c.f.u.) could be detected; a PCR product was observed from template derived from a 5 h enrichment broth culture of chicken seeded with 1 c.f.u. per gram of Enteritidis. This PCR assay is specific, reproducible, and less time consuming than the standard bacteriological methods used to detect Enteritidis. 相似文献
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TR O''Brien WA Blattner D Waters E Eyster MW Hilgartner AR Cohen N Luban A Hatzakis LM Aledort PS Rosenberg WJ Miley BL Kroner JJ Goedert 《Canadian Metallurgical Quarterly》1996,276(2):105-110
Normal women produce small amounts of active androgens. When androgen levels are elevated, such as for example in the polycystic ovary syndrome, this is followed by the development of male physical characteristics and muscle mass, structure and function as well as android adipose tissue distribution and function. Psychological features and stress reactions also seem similar to those of men. Such women have an increased risk of developing hypertension, non-insulin-dependent diabetes mellitus and cardiovascular disease. Recent data have shown that these physical, and psychological characteristics, as well as risk of ill health, are also found in the population of women selected at random. Women in the lowest quintiles of levels of sex-hormone-binding globulin--an indicator inversely related to active androgens--are at risk of developing hypertension, non-insulin-dependent diabetes mellitus and cardiovascular mortality. The mechanism probably includes muscular insulin resistance, following a relative androgen excess. It is thus apparent that androgens, even within the highest levels of the nonselected population of women, are powerful predictors of serious disease development. The population at risk might be as large as about 20% of middle-aged women. This is an area of female disease risk which requires more attention in screening and intervention procedures. 相似文献
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N6-(p-Azidobenzyl)adenosine (ABA) and nitrobenzylthioinosine (NBMPR) were employed as covalent probes of the nucleoside transport mechanism in human erythrocytes. NBMPR, a potent inhibitor of nucleoside transport, binds tightly (KD 0.3-1 nM) to specific sites on nucleoside transporter elements. ABA, a less potent inhibitor of uridine influx, competitively inhibited NBMPR binding (Ki 15 nM). [3H]ABA was bound tightly (KD 13.4 nM) but reversibly to sites on erythrocytes which appeared to be those which bind NBMPR. ABA binding was inhibited by uridine and adenosine. Irradiation with UV light caused site-bound [3H]ABA on erythrocyte membranes to become covalently bound and, similarly, photoactivation resulted in covalent attachment of membrane-bound [3H]NBMPR. In the presence of dithiothreitol, a free radical scavenger, photoactivation of the site-bound 3H-ligand on membranes depleted of extrinsic membrane proteins resulted in selective incorporation of 3H into band 4.5 of the membrane polypeptides which were resolved on sodium dodecyl sulfate-polyacrylamide gel electropherograms. This result, when considered with previous findings, indicates that the NBMPR-binding component of the nucleoside transport mechanism (or the entire mechanism, if the NBMPR site is an integral part) is a band 4.5 polypeptide. 相似文献
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A group of public health scientists from the United States and The Netherlands met at a Bicentennial Round Table Conference December 1-2, 1982, to discuss the latest developments in immunization against infectious diseases, focusing on pertussis, poliomyelitis, measles, and rubella. The major differences in immunization practices in the two countries are: (a) In The Netherlands, inactivated polio vaccine is used exclusively; in the United States, the oral polio vaccine is used. Polio-myelitis has virtually disappeared from both countries. (b) In The Netherlands, the pertussis component of DTP (diphtheria, tetanus, pertussis) is not given to children over the age of 1 year, whereas in the United States, it is given to children up to their seventh birthday. (c) Rubella vaccine is given only to girls at ages 11-12 years in The Netherlands, but to all children at ages 12-15 months in the United States. (d) Mumps vaccine is not administered to children in The Netherlands, but in the United States it is given routinely to children at 12-15 months (in combination with measles and rubella vaccine). The participants concluded that both the United States and The Netherlands have effective immunization programs that have significantly reduced the impact of these diseases. 相似文献
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The relative optical reflectance and electrical resistivity have been measured of thick films (~1μm) deposited by sputtering Al in a planar magnetron using Ar and Ar + O2 mixtures. The results obtained have a practical value in indicating the power input and related deposition rate required for given discharge conditions to obtain Al-films with a high reflectance and bulk resistivity. 相似文献