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.     

Static energy reduction techniques for microprocessor caches

Microprocessor performance has been improved by increasing the capacity of on-chip caches. However, the performance gain comes at the price of static energy consumption due to subthreshold leakage current in cache memory arrays. This paper compares three techniques for reducing static energy consumption in on-chip level-1 and level-2 caches. One technique employs low-leakage transistors in the memory cell. Another technique, power supply switching, can be used to turn off memory cells and discard their contents. A third alternative is dynamic threshold modulation, which places memory cells in a standby state that preserves cell contents. In our experiments, we explore the energy and performance tradeoffs of these techniques. We also investigate the sensitivity of microprocessor performance and energy consumption to additional cache latency caused by leakage-reduction techniques.     

The transjugular stent implantation for the treatment of malignant portal and hepatic vein obstruction in cancer patients

BACKGROUND: The increase in portal vascular resistance is a significant complication of metastatic disease to the liver or locally advanced cancer, e.g., biliary cancer. PATIENTS AND METHODS: This paper describes the successful palliative treatment of two cancer patients with portal hypertension presenting with the symptoms of tense ascites, mesenteric congestion, and severe variceal bleeding. By creating a stenttract between a hepatic vein and a main branch of the portal vein and/or by placing an extendable stent into the portal vein, the transjugular intrahepatic portosystemic stent-shunt (TIPS) technique was used to decompress the portovascular system. RESULTS: The TIPS-technique offers a new, safe and effective palliation for malignant portal hypertension. In both patients, the symptoms of the portal hypertension disappeared after the procedure. This was accompanied by a significant improvement of the patients performance status allowing an early ambulation. CONCLUSION: Our findings demonstrate the feasibility and effectiveness of the TIPS procedure as a minimal invasive treatment for portal vein decompression in selected tumor patients.     

Treatment of radiation-induced nervous system injury with heparin and warfarin

When radiation is used to treat nervous system cancer, exposure of adjacent normal nervous system tissue is unavoidable, and radiation-induced injury may occur. Acute injury is usually mild and transient, but late forms of radiation-induced nervous system injury are usually progressive and debilitating. Treatment with corticosteroids, surgery, and antioxidants is often ineffective. We treated 11 patients with late radiation-induced nervous system injuries (eight with cerebral radionecrosis, one with a myelopathy, and two with plexopathies, all unresponsive to dexamethasone and prednisone) with full anticoagulation. Some recovery of function occurred in five of the eight patients with cerebral radionecrosis, and all the patients with myelopathy or plexopathy. Anticoagulation was continued for 3 to 6 months. In one patient with cerebral radionecrosis, symptoms recurred after discontinuation of anticoagulation and disappeared again after reinstitution of treatment. We hypothesize that anticoagulation may arrest and reverse small-vessel endothelial injury--the fundamental lesion of radiation necrosis--and produce clinical improvement in some patients.     

Structural and electrophysiological analysis of annexin V mutants. Mutagenesis of human annexin V, an in vitro voltage-gated calcium channel, provides information about the structural features of the ion pathway, the voltage sensor and the ion selectivity filter

Annexin V binds to phospholipids in a calcium-dependent manner and exhibits calcium channel activity in vitro. We prepared a variety of mutants yielding information about the structure-function relationship of the ion channel activity. All mutants were characterized by X-ray crystallography, electron microscopy and electrophysiological measurements. Their structures are insignificantly changed whereas their electrophysiological properties are drastically different. Glu95, located in the central hydrophilic pore of the molecule, is crucial for the ion selectivity filter as its exchange leads to reduced calcium and increased sodium conductance. The removal of Glu17, located on the protein surface and far from the ion conduction pathway, leads to the appearance of a second conductance level of 9 pS in addition to the conductance level of about 30 pS in the wild-type molecule. This was also the case for Glu78, which is part of a weak calcium binding site. The exchange of Glu17 and Glu78 produced a mutant retaining only the smaller conductance level. We conclude that these two residues influence the angle between the two halves of the molecule, which determines the diameter of the ion conduction pathway, thereby leading to the occurrence of a second conductance level.     

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.