Identification of a neurosteroid binding site contained within the GluR2-S1S2 domain

Glutamate receptors play a major role in neural cell plasticity, growth, and maturation. The degree to which ionotropic glutamate receptors (iGluR) conduct current is dependent on binding of extracellular ligands, of which glutamate is the native agonist. Although the glutamate binding site of the GluR2 class of amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) iGluR has been structurally characterized, the allosteric sites attributed to neurosteroid binding have yet to be localized. Here, using intrinsic tryptophan fluorescence spectroscopy, we show that the extracellular glutamate binding core of the GluR2 class of AMPA receptors also binds to two neurosteroids, pregnenolone sulfate (PS) and 3α-hydroxy-5β-pregnan-20-one sulfate, both of which negatively modulate its activity. Interest in these sulfated neurosteroids stems from their differential modulation of other members of the iGluR family and their potential use as endogeneous agents for stroke therapy. In particular, whereas PS inhibits AMPA and other non-N-methyl-d-aspartate (NMDA) family members, it activates the NMDA receptor. In addition to providing evidence for binding of these neurosteroids to the glutamate binding core of the GluR2 class of AMPA receptors, our data suggests that both neurosteroids bind in a similar manner, consistent with their modulation of activity of this class of iGluR. Interestingly, the conformational change induced upon binding of these neurosteroids is distinct from that induced upon glutamate binding.     

Dynamic subset selection based on a fitness case topology

A large training set of fitness cases can critically slow down genetic programming, if no appropriate subset selection method is applied. Such a method allows an individual to be evaluated on a smaller subset of fitness cases. In this paper we suggest a new subset selection method that takes the problem structure into account, while being problem independent at the same time. In order to achieve this, information about the problem structure is acquired during evolutionary search by creating a topology (relationship) on the set of fitness cases. The topology is induced by individuals of the evolving population. This is done by increasing the strength of the relation between two fitness cases, if an individual of the population is able to solve both of them. Our new topology-based subset selection method chooses a subset, such that fitness cases in this subset are as distantly related as is possible with respect to the induced topology. We compare topology-based selection of fitness cases with dynamic subset selection and stochastic subset sampling on four different problems. On average, runs with topology-based selection show faster progress than the others.     

Bisulphite Setting of Woollen–spun Carpet Yarns

The twist in carpet yarns of suitable construction can be adequately set by a continuous treatment with sodium bisulphite in a tape–scouring machine. Different levels of set can be produced by varying the conditions: increasing the bisulphite concentration, the pH of the solution, the dwell time, and the bath temperature all improve the set. Improvements in the colour of the yarn and changes in pH are produced. While the yarn strength and extensibility decrease, the changes are comparable with those produced by the autoclave and water–setting treatments examined. Adverse changes in carpet abrasion resistance may occur when concentrations in excess of 10g/l are used. Bisulphite setting has been found to be more suitable for yarns to be jet–printed than setting in an autoclave or by hot water.     

Useful formula for moment computation of normal random variables with nonzero means

It is well known that higher order moments of normal random variables (RV) with zero means can be expressed by terms of second-order moments. In this correspondence an extension will be given for moments of normal RV with nonzero means.     

An integrated microfluidic system for reaction, high-sensitivity detection, and sorting of fluorescent cells and particles

Presented is a novel approach for an integrated micro total analysis system (microTAS) based on a microfluidic on-chip device that supports ultrasensitive confocal detection of fluorescent cells and particles and subsequently allows for their precise sorting in the fluid phase with respect to spectroscopic properties, such as brightness and color. The hybrid silicone elastomer/glass chip first comprises a branched channel system to initiate fluid mixing and to hydrodynamically focus the sample solution down to a thin flow layer, matching the size of the confocal detection volume placed at that position and, thus, providing a high detection efficiency. In the subsequent on-chip module, the dispersed cells or particles can be sorted into two different output channels. The sorting process is realized by a perpendicular deflection stream that can be switched electrokinetically. The performance of the automated sorting routine is demonstrated by precise partition of a mixture of differently colored fluorescent beads. Moreover, the specifically branched channel geometry allows for direct implementation of reaction steps prior to detection and sorting, which is demonstrated by inducing a selective recognition reaction between the fluorescent protein R-phycoerythrin and a mixture of live bacterial cells exhibiting or lacking the respective surface antigens.     

Sample preparation strategies for targeted proteomics via proteotypic peptides in human blood using liquid chromatography tandem mass spectrometry

The simultaneous quantification of protein concentrations via proteotypic peptides in human blood by liquid chromatography coupled to quadrupole MS/MS is an important field of bioanalytical research with a high potential for routine diagnostic applications. This review summarizes currently available sample preparation procedures and trends for absolute protein quantification in blood using LC-MS/MS. It discusses approaches of transferring established qualitative protocols to a quantitative analysis regarding their reliability and reproducibility. Techniques used to enhance method sensitivity such as the depletion of high-abundant proteins or the immunoaffinity enrichment of proteins and peptides are described. Furthermore, workflows for (i) protein denaturation, (ii) disulfide bridge reduction and (iii) thiol alkylation as well as (iv) enzymatic digestion for absolute protein quantification are presented. The main focus is on the tryptic digestion as a bottleneck of protein quantification via proteotypic peptides. Conclusively, requirements for a high-throughput application are discussed.     

The Influence of Disorder on Thermotropic Nematic Liquid Crystals Phase Behavior

We review the theoretical research on the influence of disorder on structure and phase behavior of condensed matter system exhibiting continuous symmetry breaking focusing on liquid crystal phase transitions. We discuss the main properties of liquid crystals as adequate systems in which several open questions with respect to the impact of disorder on universal phase and structural behavior could be explored. Main advantages of liquid crystalline materials and different experimental realizations of random field-type disorder imposed on liquid crystal phases are described.     

MIMC reliability and electrical behavior defined by a physical layer property of the dielectric

Metal–insulator–metal capacitor (MIMC) reliability and electrical properties are defined by the TDDB lifetime, breakdown voltage and leakage current. In this article, the correlation is determined between these electrical properties and the physical and chemical properties of the SiN dielectric layer. It is demonstrated how a SiN dielectrics with a high refractive index have high Si content and show an increased initial leakage current. However, contradictory to the high leakage current, these dielectrics also show high lifetimes. It is shown that SiN dielectrics with a high Si content contain high numbers of charge trapping centers. Over time, a high concentration of trapped charges is build up to such an extend that the local electric field over the dielectric is significantly decreased. This results in the observed reliability improvement of the dielectric. The final intrinsic quality and reliability of MIMC capacitors can therefore be determined by measurable physical properties of the MIMC dielectric at the time of the deposition of this layer.