Biochemical studies of the mechanism of action of the Cdc42-GTPase-activating protein

The small GTP-binding proteins Rac, Rho, and Cdc42 were shown to mediate a variety of signaling pathways including cytoskeletal rearrangements, cell-cycle progression, and transformation. Key to the proper function of these GTP-binding proteins is an efficient shut-off mechanism that ensures the decay of the signal. Regulatory proteins termed GAPs (GTPase-activating proteins) enhance the intrinsic GTP hydrolysis of the GTP-binding proteins, thereby ensuring signal termination. We have used site-specific mutagenesis to elucidate the limit domain for GAP activity in Cdc42-GAP, and show that in addition to the known GAP-homology domain (three conserved boxes), a C-terminal region outside that domain is also essential for GAP activity. In addition, we have replaced the conserved arginine (Arg305), which was suggested by structural studies to be a key catalytic residue, with an alanine and found that the R305A Cdc42-GAP mutant has a greatly diminished catalytic capacity but is still able to bind Cdc42 with high affinity. Thus, a key catalytic role for this residue is confirmed. However, we also present evidence for the involvement of an additional residue(s), since the R305A Cdc42-GAP mutant still exhibits measurable activity. Some of this residual activity might result from a neighboring arginine, since a double mutant R305A/R306A shows a further decrease in catalytic activity.     

Integrative computational approach to evaluate risk genes for postmenopausal osteoporosis

In recent years, numerous studies reported over a hundred of genes playing roles in the etiology of postmenopausal osteoporosis (PO). However, many of these candidate genes were lack of replication and results were not always consistent. Here, the authors proposed a computational workflow to curate and evaluate PO related genes. They integrate large‐scale literature knowledge data and gene expression data (PO case/control: 10/10) for the marker evaluation. Pathway enrichment, sub‐network enrichment, and gene–gene interaction analysis were conducted to study the pathogenic profile of the candidate genes, with four metrics proposed and validated for each gene. By using the authors'' approach, a scalable PO genetic database was developed; including PO related genes, diseases, pathways, and the supporting references. The PO case/control classification supported the effectiveness of the four proposed metrics, which successfully identified eight well‐studied top PO genes (e.g. TGFB1, IL6, IL1B, TNF, ESR2, IGF1, HIF1A, and COL1A1) and highlighted one recently reported PO genes (e.g. IFNG). The computational biology approach and the PO database developed in this study provide a valuable resource which may facilitate understanding the genetic profile of PO.Inspec keywords: biology computing, genetics, diseases, boneOther keywords: risk genes, postmenopausal osteoporosis, integrative computational approach, gene expression data, gene‐gene interaction analysis, IL1B, TNF, ESR2, HIF1A, COL1A1, PO genes     

Scalable stability detection using logical hypercube

This paper proposes to use a logical hypercube structure for detecting message stability in distributed systems. In particular, a stability detection protocol that uses such a superimposed logical structure is presented, and its scalability is compared with other known stability detection protocols. The main benefits of the logical hypercube approach are scalability, fault-tolerance, and refraining from overloading a single node or link in the system. These benefits become evident both by an analytical comparison and by simulations. Another important feature of the logical hypercube approach is that the performance of the protocol is in general not sensitive to the topology of the underlying physical network.     

Immune response to an intercalated enhanced inactivated polio vaccine/oral polio vaccine programme in Israel: impact on the control of poliomyelitis

A combined enhanced inactivated polio vaccine (EIPV) and oral polio vaccine (OPV) programme was introduced in Israel in 1990, with the purpose of providing a solution to the persistent polio morbidity in spite of a 30 year long OPV programme. The schedule comprised two doses of EIPV administered at the age of 2 and 4 months, intercalated with two doses of OPV at 4 and 6 months, followed by a reinforcing dose with the two vaccines simultaneously administered at 12 months. The 5-year evaluation of the programme included: the assessment of clinical suspicions of polio, early immune response in successive cohorts administered the new schedule, dynamics of the immune profile in a cohort followed up to the age of 5, and monitoring of wild poliovirus excretion in sewage specimens collected in 25 permanent sites throughout Israel as well as from the Palestinian Authority. No paralytic polio cases associated with a wild or vaccinal poliovirus strain were detected since the introduction of the programme. At the age of 4 months, one week after administration of the second EIPV and first OPV dose, 100% seropositivity and high geometric mean titres (GMTs) of neutralizing antibody (NA) to the three vaccinal and to the wild poliovirus type 1, responsible for the 1988 polio outbreak, were observed. No change in percent of seropositivity occurred between the age of 6 and 12 months. Thirty days after the IPV and OPV reinforcing doses, GMTs to each of the four poliovirus strains were > or = 3037. Up to the age of 5, the seropositivity was unchanged. After a 2.5-10-fold decline in the first year following the completion of the programme, GMTs to the three vaccinal and the wild poliovirus strain levelled off at rather high values, considered protective. Between 1990 and 1995, 16 wild poliovirus type 1 strains were isolated in three separate episodes in Gaza Strip sewage and once only in one Israeli site very close to Gaza City. The rapidly established, high and persistent NA titre to the vaccinal and wild poliovirus strains and the presence of immunological memory are indicative of high individual protection throughout the first 5 years of life. The only one-time introduction, without circulation, of a wild poliovirus strain in a single Israeli settlement suggests community protection. The intercalated programme offers a contribution to polio eradication by providing a solution to the primary and secondary failure associated with OPV, as well as to the control of vaccine-associated paralytic poliomyelitis.     

Improved neutron detection by gamma-ray spectroscopy

Most high-efficiency interactions for neutron detection are with thermal neutrons, and the detection of the prompt gamma radiation emitted as a result of thermal neutron absorption in specific materials was sometimes used. These materials are named converters, the most widespread being 10B, 113Cd or 73Ge (the last two materials are detector materials themselves for CdZnTe and HPGe detectors). However, for these materials, the main gamma-ray energies are relatively low (in the region 473-600 keV), and there are interferences from other gamma rays present in background. Different combinations of moderator-converter-detector systems were employed to optimise the system efficiency and signal-to-background ratio, using B, Cd, Gd and Cl converters and HPGe, NaI(Tl) and BGO detectors. When using a Cl converter, the 1164.7 keV peak seems to be a good choice when a HPGe spectrometer is used. A very practical material containing chlorine is PVC.     

Time-Resolved and Spatio-Temporal Analysis of Complex Cognitive Processes and their Role in Disorders like Developmental Dyscalculia

The aim of this article is to report on the importance and challenges of a time-resolved and spatio-temporal analysis of fMRI data from complex cognitive processes and associated disorders using a study on developmental dyscalculia (DD). Participants underwent fMRI while judging the incorrectness of multiplication results, and the data were analyzed using a sequence of methods, each of which progressively provided more a detailed picture of the spatio-temporal aspect of this disease. Healthy subjects and subjects with DD performed alike behaviorally though they exhibited parietal disparities using traditional voxel-based group analyses. Further and more detailed differences, however, surfaced with a time-resolved examination of the neural responses during the experiment. While performing inter-group comparisons, a third group of subjects with dyslexia (DL) but with no arithmetic difficulties was included to test the specificity of the analysis and strengthen the statistical base with overall fifty-eight subjects. Surprisingly, the analysis showed a functional dissimilarity during an initial reading phase for the group of dyslexic but otherwise normal subjects, with respect to controls, even though only numerical digits and no alphabetic characters were presented. Thus our results suggest that time-resolved multi-variate analysis of complex experimental paradigms has the ability to yield powerful new clinical insights about abnormal brain function. Similarly, a detailed compilation of aberrations in the functional cascade may have much greater potential to delineate the core processing problems in mental disorders.     

Effect of Bilayer Phospholipid Composition and Curvature on Ligand Transfer by the α-Tocopherol Transfer Protein

We report here our preliminary investigations on the mechanism of α-TTP-mediated ligand transfer as assessed using fluorescence resonance energy transfer (FRET) assays. These assays monitor the movement of the model α-tocopherol fluorescent derivative ((R)-2,5,7,8-tetramethyl-chroman-2-[9-(7-nitro-benzo[1,2,5]oxadiazol-4-yl amino)-nonyl]-chroman-6-ol; NBD-Toc) from protein to acceptor vesicles containing the fluorescence quencher TRITC-PE. We have found that α-TTP utilizes a collisional mechanism of ligand transfer requiring direct protein–membrane contact, that rates of ligand transfer are greater to more highly curved lipid vesicles, and that such rates are insensitive to the presence of anionic phospholipids in the acceptor membrane. These results point to hydrophobic features of α-TTP dominating the binding energy between protein and membrane. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. An erratum to this article can be found at     

Crystallization of 60SiO2–20MgO–10Al2O3–10BaO Glass Ceramics

Three distinctly different microstructures of silica (as quartz and crystobalite), alumina, enstatite, and celsian, were found to develop in a 60SiO₂–20MgO–10Al₂O₃–10BaO glass ceramic. At 1010°C, growth of wormy fibrillar crystals was observed, indicating that crystal growth was diffusion controlled. At the intermediate temperature of 1080°C, a coarse cellular microstructure developed with multiple spherical particles nucleated on their surfaces and in the surrounding glass. At 1200°C, the glass crystallizes in a denderitic morphology but the dendrites were actually fragmented into multiple cube-shaped enstatite crystals, indicating a transition to interface-controlled growth. The crystals coarsen with time but maintain their order along the dendrite skeletons.