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.     

Peer drug associations and emotional restraint: Causes or consequences of adolescents' drug use?

A 3-wave longitudinal design was used to examine the relationships among emotional restraint, peer drug associations, and gateway drug use in a sample of 1,256 middle school students. Structural equation modeling was used to compare 3 models: (1) One model viewed drug use as a consequence of emotional restraint and peer variables; (2) 1 viewed drug use as a cause of restraint and peer variables; and (3) 1 included reciprocal effects. All 3 models fit the data fairly well. However, the reciprocal model fit the data significantly better than either of the others. Within this model, low emotional restraint was significantly related to subsequent increases in gateway drug use among boys. In contrast, peer drug models and peer pressure were not related to subsequent changes in gateway drug use. Changes in peer drug models were, however, predicted by previous levels of gateway drug use. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The effect of topical corticosteroids on refractive outcome and corneal haze after photorefractive keratectomy

BACKGROUND: The effect of topical corticosteroids after excimer laser photorefractive keratectomy (PRK) remains a matter of some controversy. Refractive effects may be different according to the amount of myopia and timing of instillation. METHODS: Two groups of patients were studied: Study A consisted of 215 eyes (128 patients) with PRK (mean baseline myopia, -6.53 +/- 2.22 D) that received no corticosteroids (No Corticosteroid Group) unless significant regression or corneal haze appeared (Delayed Corticosteroid Group), and in Study B, we randomly assigned eyes to the Initial Corticosteroid Group (mean baseline myopia, -6.39 +/- 1.84 D) or the No/delayed Corticosteroid Group (mean baseline myopia -5.78 +/- 2.02 D). Clinical results after PRK for low-to-moderate and high myopia were compared. RESULTS: In the first group, 70.9% (73 eyes) of moderately myopic eyes (mean, -4.56 +/- 1.10 D) belonged to the No Corticosteroid Group that had a mean refraction of -5.39 +/- 1.77 D. Delayed Corticosteroid Group eyes were more myopic (mean, -7.52 +/- 2.10 D), and showed more severe haze than those in the No Corticosteroid Group. In study B, only in high myopes with more than -6.00 D (mean, -7.76 +/- 1.15 D) did refraction and corneal haze outcomes show significant difference between the Initial Corticosteroid Group and the No/delayed Corticosteroid Group. CONCLUSIONS: The effects of topical corticosteroids after PRK were less in moderate myopes compared to high myopes. Delayed instillation of corticosteroids did not reverse the regression or haze whereas initial instillation showed a beneficial effect on high myopes but not on moderate myopes.     

Sequential tripping strategy for a transmission network back-upprotection expert system

This paper describes a sequential tripping strategy used in a wide area back-up protection expert system (BPES) to combat situations in which protection relays have maloperated or information is missing. The BPES is an innovative back-up protection scheme designed to prevent the occurrence of widespread blackouts. The BPES evaluates the certainty that transmission lines are likely to be affected by the fault and uses a sequential tripping strategy to isolate the fault if a firm decision is not available due to maloperated relays and/or missing information. The mode of analysis and the sequential tripping strategy ensures that the BPES will clear a fault at minimum risk to the network. An example is included to demonstrate how the certainty factor based sequential tripping strategy is employed by the BPES to clear a fault which occurred on the South Western part of the UK National Grid System     

Investigation on Single-Mode–Multimode– Single-Mode Fiber Structure

This paper presents an investigation on a single-mode-multimode-single-mode fiber structure. A one-way guided-mode propagation analysis for the circular symmetry waveguide is employed to model the light propagation and the approximated formulations are derived and evaluated concerning the accuracy. Phase conjunction of the multimode interference within the fiber structure is revealed. A simple way to predict and analyze the spectral response of the structure is presented through the space to wavelength mapping with the derived approximated formulations. The prediction of spectral response is verified numerically and experimentally.     

Uniform and optimal schemes for stiff initial-value problems

We formulate a class of difference schemes for stiff initial-value problems, with a small parameter ε multiplying the first derivative. We derive necessary conditions for uniform convergence with respect to the small parameter ε, that is the solution of the difference scheme u_i^h satisfies |u_i^h−u(x_i)| Ch, where C is independent of h and ε. We also derive sufficient conditions for uniform convergence and show that a subclass of schemes is also optimal in the sense that |u_i^h−u(x_i)| C min (h, ε). Finally, we show that this class contains higher-order schemes.     

Larynx-associated lymphoid tissue (LALT) in young children

BACKGROUND: Mucosa-associated lymphoid tissue (MALT) plays a central role in mucosal immunity. Whereas the characteristics and function of MALT in the intestine are well established, almost nothing is known about MALT in the larynx. METHODS: In this study we examined the morphology and the lymphocyte subset composition of MALT in the larynges of children who had died of sudden infant death or various defined traumatic or nontraumatic causes. RESULTS: Organized lymphoid tissue was found in the supraglottic parts of the larynx in nearly 80% of the children in both groups. This lymphoid tissue showed all morphological signs of MALT, such as typical lymphoid follicles with germinal centers, infiltration of the overlying epithelium by lymphocytes, and high endothelial venules (HEV). Thus we will use the term LALT (larynx-associated lymphoid tissue) to refer to this tissue. The lymphoid follicles of LALT contained mainly B lymphocytes with some CD4+ lymphocytes in the germinal centers. Remarkably, T lymphocytes of both subset types and B lymphocytes were observed in comparable numbers in the parafollicular area. CONCLUSIONS: We assume that LALT is a physiological structure of the larynx in young children. The morphology and the distribution of lymphocyte subsets are similar to those of MALT in the human gut. LALT may be a regular part of the mucosal immune system in young children with the role of respiratory inductive site for mucosal immunity.     

Primary afferent depolarizations of sensory origin within contact-sensitive mechanoreceptive afferents of a crayfish leg

Recordings from the central branches of single identified dactyl sensory afferent (DSA) neurons in a crayfish in vitro preparation were performed to study modifications of the sensory message occurring before the first central synapse. These afferents comprised hairs and force-sensitive mechanoreceptors with phasic and phasotonic response characteristics in the terminal segment (dactyl) of the crayfish leg. More than one afferent spike size was often observed in intracellular recordings from these afferents, thus indicating the presence of electrical coupling between the central processes of DSA fibers. Additionally, in identified DSA fibers with large spike sizes, primary afferent depolarizations (PADs) of up to 15 mV were observed, which sometimes triggered antidromic spikes in the afferent. Nevertheless, PADs were clearly inhibitory, because they shunted the afferent spikes. They exhibited the following properties. First, each PAD was preceded by an afferent spike from a neighboring hair, indicating that the PADs had a sensory rather than central origin. Second, PADs could follow high frequencies of afferent discharges without failure, a property suggestive of monosynaptic connections, but because PAD latencies varied by +/-0.5 ms it is more likely that they were mediated by a disynaptic pathway. Third, although PADs were evoked in an extremely reliable manner, their amplitude varied in a quantal manner. Most unitary PADs were the result of the release of < 12 quanta, the mean quantal content lying between 4 and 5; quantal size was large, approximately 1 mV. Fourth, PADs showed facilitation in some fibers, whereas in others they became much smaller when occurring at brief intervals. We suggest that PADs may be an efficient and parsimonious way to limit sensory inflow in space and time, allowing the crayfish to identify precisely both weak and strong mechanical stimuli.