Detection of G proteins by affinity probe capillary electrophoresis using a fluorescently labeled GTP analogue

An affinity probe capillary electrophoresis (APCE) assay for guanine-nucleotide-binding proteins (G proteins) was developed using BODIPY FL GTPgammaS (BGTPgammaS), a fluorescently labeled GTP analogue, as the affinity probe. In the assay, BGTPgammaS was incubated with samples containing G proteins and the resulting mixtures of BGTPgammaS-G protein complexes and free BGTPgammaS were separated by capillary electrophoresis and detected with laser-induced fluorescence detection. Separations were completed in less than 30 s using 25 mM Tris, 192 mM glycine at pH 8.5 as the electrophoresis buffer and applying 555 V/cm over a 4-cm separation distance. BGTPgammaS-Galpha(o) peak heights increased linearly with Galpha(o) up to approximately 200 nM using a 50 nM BGTPgammaS probe. The detection limit for Galpha(o) was 2 nM, corresponding to a mass detection limit of 3 amol. The high speed of the APCE assays allowed reaction kinetics and the dissociation constant (Kd) to be determined. The on-rate and off-rate of BGTPgammaS to Galpha(o) were 0.0068 +/- 0.0004 and 0.000 23 +/- 0.000 01 s(-1), respectively. The half-life of the BGTPgammaS-Galpha(o) complex was 3060 +/- 240 s and Kd was 8.6 +/- 0.7 nM. The estimates of these parameters are in good agreement with those obtained using established techniques, indicating the suitability of this method for such measurements. Lowering the temperature of the separation improved the detection of the complex, allowing the assay to be performed on a commercial instrument with longer separation times. Additionally, the capability of the technique to detect several G proteins based on their binding to BGTPgammaS was demonstrated with assays for Galpha and Galpha(i1) and for Ras and Rab3A.     

Cognitive and learning difficulties and how they affect access to IT systems

In October 2005, the IBM Human Ability and Accessibility Center and T.J. Watson Research Center hosted a symposium on “cognitive and learning difficulties and how they affect access to IT systems”. The central premise of the symposium was the recognition that cognitive and learning difficulties have a profound impact on a person’s ability to interact with information technology (IT) systems, but that little support is currently being offered by those systems. By bringing together internationally renowned experts from a variety of different, but complementary, research fields, the symposium aimed to provide a complete overview of the issues related to this topic. This paper summarises the discussions and findings of the symposium.     

Integrating writing into psychotherapy practice: A matrix of change processes and structural dimensions.

Research on therapeutic writing indicates that it can offer a range of physical and psychological benefits. There is no consensus, however, concerning how writing achieves these benefits. To address this question, the authors propose a matrix framework with emotional-cognitive change processes (what can be activated) along its horizontal dimension and abstract-concrete structure (how the processes are activated) along its vertical dimension. On the horizontal dimension, writing can encourage clients who are distant from their emotional world to approach or to modulate emotional intensity, and to create meaning and coherence. Along the vertical dimension, these processes can be activated through tasks that vary in structure, including programmed writing, diaries, journaling, autobiography, storytelling, and poetry. Finally, the authors consider constraints on writing that apply to particular client groups. The matrix framework is meant to encourage clinicians to use therapeutic writing and to assist researchers in framing questions to advance our knowledge of writing as a therapeutic practice. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The role of dorsal hippocampus and basolateral amygdala NMDA receptors in the acquisition and retrieval of context and contextual fear memories.

The authors used 3-phase context preexposure facilitation methodology to study the contribution of N-methyl-D-aspartate (NMDA) receptors in dorsal hippocampus (DH) and the basal lateral region of the amygdala (BLA) to (a) acquisition of the context memory, (b) retrieval of the context memory, (c) acquisition of context-shock association, and (d) retrieval of the context-shock association. The NMDA receptor antagonist D-2-amino-5 phosphonopentanoic acid (D-AP5) was injected into either the DH or BLA prior to (a) the context preexposure phase, (b) the immediate shock phase, or (c) the test for contextual fear. Antagonizing NMDA receptors in the DH impaired the acquisition of the context memory but did not affect its retrieval or retrieval of the fear memory. Antagonizing NMDA receptors with D-AP5 in the BLA impaired acquisition of the context-shock association but had no effect on the expression of fear. However, both DL-AP5 and L-AP5 reduced the expression of fear when they were injected into the amygdala prior to testing for contextual fear. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Reciprocal relations between rumination and bulimic, substance abuse, and depressive symptoms in female adolescents.

The authors examined the reciprocal relations between rumination and symptoms of depression, bulimia, and substance abuse with longitudinal data from 496 female adolescents. Rumination predicted future increases in bulimic and substance abuse symptoms, as well as onset of major depression, binge eating, and substance abuse. Depressive and bulimic, but not substance abuse, symptoms predicted increases in rumination. Rumination did not predict increases in externalizing symptoms, providing evidence for the specificity of effects of rumination, although externalizing symptoms predicted future increases in rumination. Results suggest rumination may contribute to the etiology of depressive, bulimic, and substance abuse pathology and that the former two disturbances may foster increased rumination. Results imply that it might be beneficial for prevention programs to target this cognitive vulnerability. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Driver training conditions affect sensitivity to the impairing effects of alcohol on a simulated driving test to the impairing effects of alcohol on a simulated driving test.

[Correction Notice: An erratum for this article was reported in Vol 16(2) of Experimental and Clinical Psychopharmacology (see record 2008-03846-009). The correct title of the article should read "Driver training conditions affect sensitivity to the impairing effects of alcohol on a simulated driving test".] Research shows that prior behavioral training in a challenging environment reduces alcohol-induced impairment on simple psychomotor tasks. However, no studies have examined if this relationship generalizes to driving performance. The present study examined simulated driving performance and tested the hypothesis that a challenging training history would protect against the impairing effects of alcohol on driving performance. The challenging training history involved driving in a visually-impoverished environment. Thirty adults were randomly assigned to one of three groups. Two groups were tested under alcohol (0.65 g/kg) after prior experience performing the task under either a visually-impoverished environment or a normal visual environment. The remaining group served as a control and was trained and tested under the visually-impoverished condition environment. Results showed that individuals trained in the impoverished environment displayed sober levels of performance when their performance was subsequently tested under alcohol. By contrast, volunteers trained in a normal environment showed impairment under alcohol. The findings suggest that differences in driving training history can affect a driver's sensitivity to the impairing effects of alcohol. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

In situ synthesized ceramic–polymer composites for bone tissue engineering: bioactivity and degradation studies

As an alternative to current bone grafting strategies, a poly-lactide-co-glycolide/calcium phosphate composite microsphere-based scaffold has been synthesized by the direct formation of calcium phosphate within forming microspheres. It was hypothesized that the synthesis of low crystalline calcium phosphate within forming microspheres would provide a site-specific delivery of calcium ions to enhance calcium phosphate reprecipitation onto the scaffold. Both polymeric and composite scaffolds were incubated in simulated body fluid (SBF) for 8 weeks, during which time polymer molecular weight, scaffold mass, calcium ion concentration of SBF, pH of SBF, and calcium phosphate reprecipitation was monitored. Results showed a 20% decrease in polymeric scaffold molecular weight compared to 11–14% decrease for composite scaffolds over 8 weeks. Composite scaffold mass and SBF pH decreased for the first 2 weeks but began increasing after 2 weeks and continued to do so up to 8 weeks, suggesting interplay between pH changes and calcium phosphate dissolution/reprecipitation. Free calcium ion concentration of SBF containing composite scaffolds increased 20–40% over control values within 4 h of incubation but then dropped as low as 40% below control values, suggesting an initial burst release of calcium ions followed by a reprecipitation onto the scaffold surface. Scanning electron micrographs confirm calcium phosphate reprecipitation on the scaffold surface after only 3 days of incubation. Results suggest the composite scaffold is capable of initiating calcium phosphate reprecipitation which may aid in bone/implant integration.     

Daunorubicin‐Loaded DNA Origami Nanostructures Circumvent Drug‐Resistance Mechanisms in a Leukemia Model

Many cancers show primary or acquired drug resistance due to the overexpression of efflux pumps. A novel mechanism to circumvent this is to integrate drugs, such as anthracycline antibiotics, with nanoparticle delivery vehicles that can bypass intrinsic tumor drug‐resistance mechanisms. DNA nanoparticles serve as an efficient binding platform for intercalating drugs (e.g., anthracyclines doxorubicin and daunorubicin, which are widely used to treat acute leukemias) and enable precise structure design and chemical modifications, for example, for incorporating targeting capabilities. Here, DNA nanostructures are utilized to circumvent daunorubicin drug resistance at clinically relevant doses in a leukemia cell line model. The fabrication of a rod‐like DNA origami drug carrier is reported that can be controllably loaded with daunorubicin. It is further directly verified that nanostructure‐mediated daunorubicin delivery leads to increased drug entry and retention in cells relative to free daunorubicin at equal concentrations, which yields significantly enhanced drug efficacy. Our results indicate that DNA origami nanostructures can circumvent efflux‐pump‐mediated drug resistance in leukemia cells at clinically relevant drug concentrations and provide a robust DNA nanostructure design that could be implemented in a wide range of cellular applications due to its remarkably fast self‐assembly (≈5 min) and excellent stability in cell culture conditions.     

Inhibition of hIAPP Amyloid Aggregation and Pancreatic β‐Cell Toxicity by OH‐Terminated PAMAM Dendrimer

Human islet amyloid polypeptide (hIAPP, or amylin) forms amyloid deposits in the islets of Langerhans, a phenomenon that is associated with type‐2 diabetes impacting millions of people worldwide. Accordingly, strategies against hIAPP aggregation are essential for the prevention and eventual treatment of the disease. Here, it is shown that generation‐3 OH‐terminated poly(amidoamine) dendrimer, a polymeric nanoparticle, can effectively halt the aggregation of hIAPP and shut down hIAPP toxicity in pancreatic MIN6 and NIT‐1 cells as well as in mouse islets. This finding is supported by high‐throughput dynamic light scattering experiment and thioflavin T assay, where the rapid evolution of hIAPP nucleation and elongation processes is halted by the addition of the dendrimer up to 8 h. Discrete molecular dynamics simulations further reveal that hIAPP residues bound strongly with the dendrimer near the c‐terminal portion of the peptide, where the amyloidogenic sequence (residues 22–29) locates. Furthermore, simulations of hIAPP dimerization reveal that binding with the dendrimer significantly reduces formation of interpeptide contacts and hydrogen bonds, thereby prohibiting peptide self‐association and amyloidosis. This study points to a promising nanomedicinal strategy for combating type‐2 diabetes and may have broader implications for targeting neurological disorders whose distinct hallmark is also amyloid fibrillation.     

Do problematic and non-problematic video game players differ in extraversion,trait empathy,social capital and prosocial tendencies?

The increasing popularity of online and multiplayer games has meant that for many, social interaction and cooperation are vital components of the gaming experience. Previous research has suggested that not only has this made gaming more attractive to socially oriented people but also that it may be socially beneficial in terms of social capital and prosocial behaviors. However, for problematic video game players (those showing signs of compulsive or detrimental video game use), this may not be the case, and a number of theories hold deficiencies in socializing in real life as central to the development of this issue. In the present study, an online questionnaire completed by 416 participants assessed problematic video game use, extraversion, trait empathy, online and offline social capital and prosocial tendencies. Contrary to hypotheses, non-problematic, problematic and non-gamers did not differ in empathy, extraversion or prosocial tendencies. Problematic video game play was, however, associated with significantly higher online social capital and lower offline social capital whereas non-problematic players demonstrated only higher online capital than non-players. This highlights the importance of social support but suggests personality is not an influential factor.