Purification of the cardiac sarcoplasmic reticulum membrane protein phospholamban from recombinant Escherichia coli

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.     

Scarring trachoma is associated with polymorphism in the tumor necrosis factor alpha (TNF-alpha) gene promoter and with elevated TNF-alpha levels in tear fluid

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.     

A delta transform approach to loop gain-phase shaping design of robust digital control systems

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.     

Optimal ensemble construction via meta-evolutionary ensembles

In this paper, we propose a meta-evolutionary approach to improve on the performance of individual classifiers. In the proposed system, individual classifiers evolve, competing to correctly classify test points, and are given extra rewards for getting difficult points right. Ensembles consisting of multiple classifiers also compete for member classifiers, and are rewarded based on their predictive performance. In this way we aim to build small-sized optimal ensembles rather than form large-sized ensembles of individually-optimized classifiers. Experimental results on 15 data sets suggest that our algorithms can generate ensembles that are more effective than single classifiers and traditional ensemble methods.     

The effects of recent practice on task switching.

Four experiments investigated the effect of recent selective practice on the cost of switching between 2 tasks afforded by letter-digit pairs: alphabet arithmetic and shape comparison. Experiments 1 and 2 found a greater cost associated with switching to the more recently practiced task: evidence that task-set inertia contributes to switching costs. Experiment 3 found this effect to be limited to trials on which a recently trained stimulus followed another such stimulus: a result problematic for all current theories of task-set priming. Experiment 4 showed that the effect of recent practice was eliminated by active preparation for a task switch: It appears that endogenous task-set preparation reduces the effects of task-set inertia. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Equivalence of models for polynomial learnability

In this paper we consider several variants of Valiant's learnability model that have appeared in the literature. We give conditions under which these models are equivalent in terms of the polynomially learnable concept classes they define. These equivalences allow comparisons of most of the existing theorems in Valiant-style learnability and show that several simplifying assumptions on polynomial learning algorithms can be made without loss of generality. We also give a useful reduction of learning problems to the problem of finding consistent hypotheses, and give comparisons and equivalences between Valiant's model and the prediction learning models of Haussler, Littlestone, and Warmuth (in “29th Annual IEEE Symposium on Foundations of Computer Science,” 1988).