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41.
We have used a variety of methods to characterize the genome of the archaeon Methanosarcina thermophila TM-1. Pulsed-field gel analysis indicates a genome size of 2.8 Mb. We have constructed a bacterial artificial chromosome (BAC) library of M. thermophila and have used it to generate physical maps for this organism. The library is made up of 384 clones with an average insert size of 58 kb representing 8.0 genome equivalents. The utility of the library for low-resolution physical mapping was shown by identifying NotI linking clones and using these to order the NotI macrorestriction fragments of M. thermophila into a 2.8 Mb map. Hybridization of nine single copy genes and a 16S rRNA sequence to these macrorestriction fragments forms the basis for the first genetic map in this organism. High-resolution physical maps, consisting of overlapping clones, have been created using HindIII fingerprints of BAC clones. In this way, we identified a minimal path of five clones that span a 270 kb NotI fragment. The ease of manipulating BAC clones makes the BAC system an excellent choice for the construction of low-resolution and high-resolution physical and genetic maps of archaeal genomes. It also provides a substrate for future genome-sequencing efforts.  相似文献   
42.
Finite element simulations of the high-temperature behavior of single-phase γ, dual-phase α2+γ, and fully lamellar (FL) α2+γTiAl intermetallic alloy microstructures have been performed. Nonlinear viscous primary creep deformation is modeled in each phase based on published creep data. Models were also developed that incorporate grain boundary and lath boundary sliding in addition to the dislocation creep flow within each phase. Overall strain rates are compared to gain an understanding of the relative influence each of these localized deformation mechanisms has on the creep strength of the microstructures considered. Facet stress enhancement factors were also determined for the transverse grain facets in each model to examine the relative susceptibility to creep damage. The results indicate that a mechanism for unrestricted sliding of γ lath boundaries theorized by Hazzledine and co-workers leads to unrealistically high strain rates. However, the results also suggest that the greater creep strength observed experimentally for the lamellar microstructure is primarily due to inhibited former grain boundary sliding (GBS) in this microstructure compared to relatively unimpeded GBS in the equiaxed microstructures. The serrated nature of the former grain boundaries generally observed for lamellar TiAl alloys is consistent with this finding.  相似文献   
43.
Factor V Leiden mutation was initially detected in thrombophilic patients and relatives by PCR RFLP (Restriction Fragment Length Polymorphism) according to Bertina (1). This technique presents some drawbacks and the current trend is to simplify the diagnosis. We describe a technique of Allele Specific Amplification (ASA) which is optimized in terms of reliability: an additional mismatch in antepenultimate position enables to obtain the same specificity as PCR RFLP. Furthermore, coamplification of internal control warrants an optimal sensitivity. All the PCR have been simplified: the DNA extraction improvement allows to analyse the genotype with only a few microliters of whole blood whatever the anticoagulant and the procedure of preservation (freezing, dried blood spots, storage at +4 degrees C for several days). This technique saves time. Moreover, full automation of the ASA technique may be shortened thanks to the lack of extraction and the positive/negative reading of the PCR signal.  相似文献   
44.
The road to employee empowerment runs between true commitment on the one side and enforced compliance and chaos on the other. By following this column's guidelines, IS managers can steer their organizations to success and help free employees to be more creative in terms of both quality and productivity.  相似文献   
45.
We recently reported that epidermal immunization using the PowderJet particle delivery device with plasmid vector pcDNA1/EDIM6 encoding rotavirus VP6 of murine strain EDIM induced high levels of serum rotavirus IgG but failed to protect mice against EDIM infection (Choi, A. H., Knowlton, D. R., McNeal, M. M., and Ward, R. L. (1997) Virology 232, 129-138.). This was extended to determine whether pcDNA1/EDIM4 or pcDNA1/EDIM7, which encode either rotavirus VP4 or VP7, the rotavirus neutralization proteins, could also induce rotavirus-specific antibody responses and if these responses resulted in protection. Titers of rotavirus serum IgG increased with the first dose in mice immunized with pcDNA1/EDIM7, but little or no serum rotavirus IgG was detected in mice immunized with pcDNA1/EDIM4. In vitro assays with these plasmids in rabbit reticulocyte lysates showed that VP4 was expressed but the amount was considerably lower than VP6 or VP7. To improve expression of VP4 and induction of rotavirus-specific humoral responses, the coding region of VP4 was cloned into the high-expression plasmid WRG7054 as a fusion protein containing the 22-amino-acid secretory signal peptide of tissue plasminogen activator (tPA) at its N terminus. In vitro expression of tPA::VP4 was significantly higher than unmodified VP4, and mice inoculated with WRG7054/EDIM4 generated high titers of rotavirus IgG. The coding sequence of VP7 without the first 162 nucleotides was also cloned into WRG7054, but no difference was observed between titers of serum rotavirus IgG in mice immunized with this plasmid (WRG7054/EDIM7Delta1-162) and pcDNA1/EDIM7. The rotavirus-specific IgG titers in all immune sera were predominantly IgG1 indicating induction of Th 2-type responses. None of the mice immunized with any of the VP4 or VP7 plasmids developed serum or fecal rotavirus IgA or neutralizing antibody to EDIM. When immunized mice were challenged with EDIM virus, there was no significant reduction in viral shedding relative to unimmunized controls. Therefore epidermal immunization with VP4 or VP7 alone elicited rotavirus IgG responses but did not protect against homologous rotavirus challenge.  相似文献   
46.
Helical distributions of stokeslets can valuably model microbial locomotion through a fluid, and also the flow field generated, wherever a flagellum actively executes helical undulations (as in many single-celled algae and protozoa) or where (as in many bacteria) the action of rotary motors causes a passive structure of helical shape (which may be a flagellum or else the cell body itself) to rotate. Here, previous biomechanical studies of such modes of locomotion are extended to include analyses of three-dimensional flow fields. In some cases, a rotlet field (curl of a stokeslet) needs to be incorporated in the models. For example, spirochete swimming is modelled by combined helical distributions of stokeslets and rotlets; the computed flow field being confined to within distances of less than twice the radius of the cell body's helical shape from its axis, while including a powerful jet-like interior flow through the coils of the swimming spirochete.  相似文献   
47.
Lorentz [1] pioneered the representation of flows at very low Reynolds number by a surface distribution of stokeslets — whose strengths, nowadays, are computed by surface-velocity collocations. That method is here compared with a representation widely used in flagellar hydrodynamics, by a curvilinear distribution of stokeslets and dipoles along the flagellar centreline; with the velocity of each cross-section expressed as a centreline value of the combined fields of singularities beyond a certain cutoff distance. The latter is also a good representation, and offers moreover some computational advantages. This paper establishes the equivalence of the two representations, and identifies those properties of Stokes flows which make both the dipoles and the cutoff essential to that equivalence.  相似文献   
48.
OBJECTIVE: To determine the accuracy of patient recall of a question about smoking in a specified consultation in general practice; the accuracy of smokers' recall of advice to stop smoking; and predictors of accurate recall. DESIGN: Analysis of 1075 audiotapes was compared blind with matching patient questionnaires administered after the consultation to calculate sensitivity and specificity for patient recall as a dichotomous variable. Predictors of recall were determined by logistic regression. SETTING: General practitioner training practices, New South Wales, Australia. SUBJECTS: Thirty-four trainees and consecutive samples of their patients 16-65 years of age. RESULTS: Patient recall of a question about smoking had a high false positive rate of 21% but a sensitivity of 93%. Smokers over-reported advice to stop smoking (specificity 82%; sensitivity 92%). Predictors of accuracy of a question about smoking included patient sex (women 1.58 times more likely than men to be accurate); smoking status (smokers 1.7 times as likely as non-smokers to be accurate); and interval since consultation (those who completed their questionnaires within a week were 1.84 times more likely to be accurate). CONCLUSIONS: Patient recall is systematically biased towards over-reporting of a question about smoking status and, among smokers, of advice to quit. Although we recommend its continued application in health services evaluation, findings should be interpreted with caution, particularly if subjects are male or the interval between recall of smoking cessation advice and the occasion of service in which it might have occurred is considerably delayed.


  相似文献   
49.
Treatment of the addictions is changing. Psychosocial treatment programs and research projects have changed the way helping professionals view, treat, and prevent drug abuse. This article reviews the contributions included in this special series; they encompass several general issues facing psychologists who conduct treatment, research, and teaching in the field of addictive behaviors. (PsycINFO Database Record (c) 2010 APA, all rights reserved)  相似文献   
50.
Abstract: Environmental justice advocates have recently focused attention on cumulative exposure in minority neighborhoods due to multiple sources of pollution. This article uses U.S. EPA's National Air Toxics Assessment (NATA) for 1996 to examine environmental inequality in California, a state that has been a recent innovator in environmental justice policy. We first estimate potential lifetime cancer risks from mobile and stationary sources. We then consider the distribution of these risks using both simple comparisons and a multivariate model in which we control for income, land use, and other explanatory factors, as well as spatial correlation. We find large racial disparities in California's “riskscape” as well as inequalities by other factors and suggest several implications for environmental and land use policy.  相似文献   
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