Proton-Detected Solid-State NMR of the Cell-Free Synthesized α-Helical Transmembrane Protein NS4B from Hepatitis C Virus

Proton-detected 100 kHz magic-angle-spinning (MAS) solid-state NMR is an emerging analysis method for proteins with only hundreds of microgram quantities, and thus allows structural investigation of eukaryotic membrane proteins. This is the case for the cell-free synthesized hepatitis C virus (HCV) nonstructural membrane protein 4B (NS4B). We demonstrate NS4B sample optimization using fast reconstitution schemes that enable lipid-environment screening directly by NMR. 2D spectra and relaxation properties guide the choice of the best sample preparation to record 2D ¹H-detected ¹H,¹⁵N and 3D ¹H,¹³C,¹⁵N correlation experiments with linewidths and sensitivity suitable to initiate sequential assignments. Amino-acid-selectively labeled NS4B can be readily obtained using cell-free synthesis, opening the door to combinatorial labeling approaches which should enable structural studies.     

On keys' meanings and modes: the impact of different key solutions on children's efficiency using a mobile phone

The present study investigates the impact of different key solutions of mobile phones on users' effectiveness and efficiency using the devices. In the first experiment, 36 children (9 - 14 years) and in the second experiment 45 young adults (19 - 33 years) completed four common phone tasks twice consecutively on three simulated phones that had identical menus, but different key solutions. An approach was undertaken to quantify the complexity of keys in three models, incorporating different factors contributing to the keys' complexity (number of key options, number of modes and number of modes with a semantically dissimilar meaning), in order to predict users' performance decrements. As a further main factor, the degree of the users' locus of control (LOC) was measured and interactions with performance outcomes were studied. As dependent measures, the number of inefficient keystrokes, the number of tasks solved and the processing time were determined. Results showed a significant effect of control key solutions on users' efficiency and effectiveness for both children and young adults. Moreover, children's LOC values significantly interacted with performance: children with low LOC values showed the lowest performance and no learnability, especially when using keys with a high complexity. From the three factors contributing to the complexity of keys, keys exerting different functions with semantically inconsistent meanings had the worst effect on performance. It is concluded that in mobile user interface design keys with semantically inconsistent meanings should be generally avoided.     

Differential effects of cue changes and task changes on task-set selection costs.

A task-switching paradigm with a 2:1 mapping between cues and tasks was used to separate cue-switching processes (indexed through pure cue-switch costs) from actual task-switching processes (indexed through additional costs in case of cue and task changes). A large portion of total switch costs was due to cue changes (Experiments 1 and 2), and cue-switch costs but not task-switch costs were sensitive to effects of practice (Experiment 1) and preparation (Experiment 2). In contrast, task-switch costs were particularly sensitive to response-priming effects (Experiments 1 and 2) and task-set inhibition (Experiment 3). Results suggest two processing stages relevant during task-set selection: cue-driven retrieval of task rules from long-term memory and the automatic application of rules to a particular stimulus situation. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

In vitro cytotoxicity and DNA damage production in Chinese hamster ovary cells and topoisomerase II inhibition by 2-[2''-(dimethylamino)ethyl]-1, 2-dihydro-3H-dibenz[de,h]isoquinoline-1,3-diones with substitutions at the 6 and 7 positions (azonafides)

The p53 tumor suppressor gene encodes a phosphoprotein which when overexpressed can induce growth arrest at the G1 and G2/M phases of the cell cycle, promote differentiation and apoptosis. This paper demonstrates that p53 can associate with trk tyrosine kinase. Expression of a murine temperature-sensitive (ts) p53 mutant in PC12 cells overexpressing trk (a model system to analyse cellular differentiation and signal transduction induced by NGF) induces morphological changes in the absence of NGF stimulation at 32 degrees C but not at 37 degrees C. In cells differentiated by p53, trk, but not EGFr, was hyperphosphorylated on tyrosine. Furthermore trk was not phosphorylated when expressed in Saos-2 cells (human osteosarcoma cells that lack expression of both endogenous trk and p53) at either temperature. However, transfection of ts p53 into these cells induces trk phosphorylation at 32 degrees C in the absence of NGF stimulation. Association of trk and p53 can be detected in NIH3T3 and PC12 cells co-expressing trk and the ts p53 mutant, in NIH3T3 and PC12 cells transfected with trk alone, and in untransfected PC12 cells, showing that overexpressed and/or endogenous trk associates with endogenous, low levels of p53. These data suggest a novel function for p53 which involves the stimulation of signal transduction pathways (mediating morphological properties of cells), possibly through association with and hyperphosphorylation of trk.     

A Study of Integrated Document and Connection Caching in the WWW

Document caching and connection caching are extensively studied problems. In document caching, one has to maintain caches containing documents accessible in a network. In connection caching, one has to maintain a set of open network connections that handle data transfer. Previous work investigated these two problems separately while in practice the problems occur together: In order to load a document, one has to establish a connection between network nodes if the required connection is not already open. In this paper we present the first study that integrates document and connection caching. We first consider a very basic model in which all documents have the same size and the cost of loading a document or establishing a connection is equal to 1. We present deterministic and randomized online algorithms that achieve nearly optimal competitive ratios unless the size of the connection cache is extremely small. We then consider general settings where documents have varying sizes. We investigate a FAULT model in which the loading cost of a document is 1 as well as a BIT model in which the loading cost is equal to the size of the document.