Validation of a new enzyme-linked immunosorbent assay to detect the triggering proteins and peptides for celiac disease: interlaboratory study

The performance of Gluten-Tec (EuroProxima, Arnhem, The Netherlands) was tested through an interlaboratory study in accordance with AOAC guidelines. Gluten-Tec is a competitive ELISA that detects an immunostimulatory epitope of a-gliadin in dietary food for celiacs. Fifteen laboratories, representing 14 different countries, announced their interest in taking part in this study. Of the 12 laboratories that sent the results within the established timeframe, two submitted inappropriate standard curves and were excluded from the statistical analysis. Four different food matrixes (rice-based baby food, maize bread, chocolate cake mix, and beer) were selected for preparing the test samples. Two gliadin extraction procedures were used: the conventional 60% ethanol, and a new method based on the reducing reagent dithiothreitol. The 38 samples (19 blind duplicates) tested in this study were prepared by diluting the different extracts in order to cover a wide range of gliadin levels. Both sample extraction and dilution were performed by EuroProxima; the present interlaboratory study was focused only on testing the ELISA part of the Gluten-Tec kit protocol. Repeatability values (within-laboratory variance), expressed as RSD(r) ranged from 6.2 to 25.7%, while reproducibility values (interlaboratory variance), expressed as RSD(R), ranged from 10.6 to 45.9%. Both statistical parameters were in the acceptable range of ELISAs under these conditions, and the method will be presented to the Codex Alimentarius as a preferred method for gluten analysis.     

Reactivity of amino acid nucleoside phosphoramidates: a mechanistic quantum chemical study

Recent experimental evidence (Maiti et al. Chem.-Eur. J., submitted) indicates that hydrolysis of nucleoside phosphoramidates is subjected to anchimeric influence by carboxyl moieties in the leaving group but also by the base in the nucleotide. A quantum chemical analysis of these findings is presented. First the intrinsic hydrolysis mechanism is investigated for simplified model compounds, and then both amino acid and nucleoside substituents are included. It is found that hydrolysis is assisted by the α-carboxyl group via formation of a five-membered intermediate and that the barrier for the reaction of this intermediate toward the product state can be influenced by the nucleobase. The adenine base protonated on N3 interacts with the transition state and considerably lowers the barrier for hydrolysis. The influence of several base modifications is explained by calculating the pK(a) for protonation on N3.     

Speeding up Martins’ algorithm for multiple objective shortest path problems

The latest transportation systems require the best routes in a large network with respect to multiple objectives simultaneously to be calculated in a very short time. The label setting algorithm of Martins efficiently finds this set of Pareto optimal paths, but sometimes tends to be slow, especially for large networks such as transportation networks. In this article we investigate a number of speedup measures, resulting in new algorithms. It is shown that the calculation time to find the Pareto optimal set can be reduced considerably. Moreover, it is mathematically proven that these algorithms still produce the Pareto optimal set of paths.     

Synthesis of modified peptidoglycan precursor analogues for the inhibition of glycosyltransferase

The peptidoglycan glycosyltransferases (GTs) are essential enzymes that catalyze the polymerization of glycan chains of the bacterial cell wall from lipid II and thus constitute a validated antibacterial target. Their enzymatic cavity is composed of a donor site for the growing glycan chain (where the inhibitor moenomycin binds) and an acceptor site for lipid II substrate. In order to find lead inhibitors able to fill this large active site, we have synthesized a series of substrate analogues of lipid I and lipid II with variations in the lipid, the pyrophosphate, and the peptide moieties and evaluated their biological effect on the GT activity of E. coli PBP1b and their antibacterial potential. We found several compounds able to inhibit the GT activity in vitro and cause growth defect in Bacillus subtilis . The more active was C16-phosphoglycerate-MurNAc-(L-Ala-D-Glu)-GlcNAc, which also showed antibacterial activity. These molecules are promising leads for the design of new antibacterial GT inhibitors.     

Electrospun polystyrene fibers for HIV entrapment

The high versatility and ease of electrospinning of polymer solutions have recently resulted in electrospun fibers, which are of interest for a wide variety of chemical and biomedical applications. This is partially due to the high surface area of the fibers, which is attractive for the detection and capture of (bio)chemicals. In the present work, polystyrene (PS) fibers were electrospun and coated with cationic poly(allylamine hydrochloride) (PAH) or anionic dextran sulfate sodium (DSS). The fibers were physicochemically characterized. Upon incubation in a dispersion of inactivated HIV‐1, avid binding of HIV to all types of fibers occurred. By atomic force microscopy and spatial selective photobleaching, the binding of the inactivated HIV‐1 particles to the fibers could be confirmed. Interestingly, all fibers, especially the DSS‐coated and PAH‐coated ones, resulted in a significant reduction of infection of CD4⁺ TZMbl cells by replication‐competent HIV‐1. On top, DSS‐coated PS fibers were not toxic for vaginal epithelial cells, which may make these fibers of potential interest to inhibit HIV infection in the context of topical prevention. Copyright © 2014 John Wiley & Sons, Ltd.     

Properties of the Interfacial Transition Zone in Model Concrete

The interfacial transition zone (ITZ) between aggregates and cement paste in cementitious materials is a crucial element in mechanical and transport systems. Computer simulation by the SPACE system is used to approach this problem in the present paper. For the particle-packing phenomenon in the fresh state of concrete, the SPACE system relies on a dynamic generation algorithm, reflecting the production conditions of concrete. Hence, structure of the model cement has been proven more realistic than can be achieved by random generator-based system.A natural phenomenon in the ITZ around aggregate particles is size segregation leading to different gradients in porosity, particle size and surface area. Size segregation implies the difference size fractions in the binder mixture to have peak values in their densities on different distances from the aggregate surface. Structural evolution of the ITZ is stereologically quantified with the help of composition and configuration parameters in the fresh and hardened states of concrete.The addition of mineral admixtures is a successful approach to improving the ITZ microstructure. Experiments demonstrated the blending efficiency to be higher for coarser grained Portland cement (PC), due to the positive effect exerted by gap grading, i.e. by having distinctly different size ranges of particles. This is confirmed by computer simulation of the ITZ microstructure in model concretes made with blended cements. In addition, it is very important in concrete production to achieve good workability conditions to ensure sufficient dispersion of the fine rice husk ash (RHA) particles and proper migration of them into the ITZ through the structure network of large cement particles.