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Phospholamban (PLN) was expressed in Escherichia coli as a protein fusion with glutathione S-transferase (GST). GST-PLN was mostly present in the insoluble protein fraction and accounted for approximately 50% of total insoluble protein. Attempts to suppress inclusion body formation or to use GST as an affinity-purification tag failed. A successful purification method is based on preparative SDS/PAGE and electrodialysis. From 1 g cells we typically purified 13.5 mg fusion protein with a PLN content of 2.8 mg. We genetically inserted an enterokinase (EK) protease site just in front of the PLN sequence and demonstrated the proteolytical liberation of PLN from the carrier protein. The approach described represents a substantial advancement in PLN expression and purification.  相似文献   
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Tumor necrosis factor alpha (TNF-alpha) may play a central role in the disease pathogenesis which occurs as a consequence of chlamydial infection. To investigate the importance of TNF-alpha gene promoter polymorphisms and TNF-alpha levels in tear fluid in scarring trachoma, a large matched-pair case-control study was performed in The Gambia. The -308A allele was present in a higher proportion of patients (28.4%) than controls (18.4%), with an increasing association for homozygotes (chi2 for trend, P = 0.032; allele frequency, 0.163 in patients and 0.099 in controls; chi2, P = 0.025). For the -238A allele, the association was similar but not significant. The disease association was highly significant when the number of either -308A or -238A sites in an individual was considered (P = 0.003). TNF-alpha promoter alleles are tightly linked to some HLA class I and II alleles, but multivariate analysis confirmed that the disease associations were independent of HLA, although a class I allele, A*6802, is also associated with disease. TNF-alpha was more frequently detected in tear samples from patients (27.6%) than from controls (15.9%), increasingly so for higher levels of detectable TNF-alpha (P = 0.015). Among patients, detectable TNF-alpha in tears was highly associated with the presence of ocular chlamydial infection (P < 0.001). The results indicate that TNF-alpha plays a major role in the tissue damage and scarring which occurs as a consequence of Chlamydia trachomatis infection.  相似文献   
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This paper addresses the existence of loop gain-phase shaping (LGPS) solutions for the design of robust digital control systems for SISO, minimum-phase, continuous-time processes with parametric uncertainty. We develop the frequency response properties of LGPS for discrete-time systems using the Δ-transform, a transform method that applies to both continuous-time and discrete-time systems. A theorem is presented which demonstrates that for reasonable specifications there always exists a sampling period such that the robust digital control problem has a solution. Finally, we offer a procedure for estimating the maximum feasible sampling period for LGPS solutions to robust digital control problems.  相似文献   
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Computer-aided software engineering ( CASE) is changing the way IS professionals develop information systems. This article demonstrates how traditional systems development methods are changing with automation, describes a new six-phase software development methodology— the CASE life cycle— designed to maximize the effective use of CASE technologies, and discusses how, organizations can make the transition to this new methodology.  相似文献   
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Examined the theory that chronic migraine headaches can lead to permanent changes in brain function in 71 female college students. Ss were 20 classic migraineurs, 27 common migraineurs, and 24 controls. No significant group differences were found on the Halstead Reitan Neuropsychological Test Battery and additional memory tests. Classic migraineurs had significantly higher scores than common migraineurs on the Hysteria and Paranoia scales of the Minnesota Multiphasic Personality Inventory (MMPI) and significantly higher scores than the controls on Hypochondriasis and Hysteria scales. Explanations for the lack of cognitive deficits in this group are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)  相似文献   
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Cross-wire anemometry was used to identify and characterize coherent flow pulsations in isothermal air flow near the gap regions of a five-rod bundle with a design pitch-to-diameter ratio of 1.149 and contained in a quasi-trapezoidal duct. It was confirmed that such pulsations are quasi-periodic and contribute significantly to the velocity fluctuations across the gap. The frequency of pulsations was found to decrease with diminishing rod–wall gap size in the range between 0.015D and 0.250D, where D is the rod diameter. The pulsations in a rod–wall gap and an adjacent rod–rod gap were strongly coupled and occurred at the same frequency as one rod was displaced towards the duct wall.  相似文献   
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