Design and Characterization of a New pVII Combinatorial Phage Display Peptide Library for Protease Substrate Mining Using Factor VII Activating Protease (FSAP) as Model

We describe a novel, easy and efficient combinatorial phage display peptide substrate-mining method to map the substrate specificity of proteases. The peptide library is displayed on the pVII capsid of the M13 bacteriophage, which renders pIII necessary for infectivity and efficient retrieval, in an unmodified state. As capture module, the 3XFLAG was chosen due to its very high binding efficiency to anti-FLAG mAbs and its independency of any post-translational modification. This library was tested with Factor-VII activating protease (WT-FSAP) and its single-nucleotide polymorphism variant Marburg-I (MI)-FSAP. The WT-FSAP results confirmed the previously reported Arg/Lys centered FSAP cleavage site consensus as dominant, as well as reinforcing MI-FSAP as a loss-of-function mutant. Surprisingly, rare substrate clones devoid of basic amino acids were also identified. Indeed one of these peptides was cleaved as free peptide, thus suggesting a broader range of WT-FSAP substrates than previously anticipated.     

Removal of nitrates from groundwater through the use of artificial wetlands

Intensive fertilizer use has led to nitrate contamination of groundwater in many areas. This article examines the possibility of removing nitrates through the construction of artificial wetlands. A pilot study conducted in Denmark has found that the method is effective and economical, at least for small waterworks, though further treatment is necessary before an acceptable microbiological water quality is attained.     

Effective modelling of percolation at the landscape scale using data-based approaches

Process-based models have been extensively applied to assess the impact of landuse change on water quantity and quality at landscape scales. However, the routine application of those models suffers from large computational efforts, lack of transparency and the requirement of many input parameters. Data-based models such as Feed-Forward Multilayer Perceptrons (MLP) and Classification and Regression Trees (CART) may be used as effective models, i.e. simple approximations of complex process-based models. These data-based approaches can subsequently be applied for scenario analysis and as a transparent management tool provided climatic boundary conditions and the basic model assumptions of the process-based models do not change dramatically. In this study, we apply MLP, CART and Multiple Linear Regression (LR) to model the spatially distributed and spatially aggregated percolation in soils using weather, groundwater and soil data. The percolation data is obtained via numerical experiments with Hydrus1D. Thus, the complex process-based model is approximated using simpler data-based approaches. The MLP model explains most of the percolation variance in time and space without using any soil information. This reflects the effective dimensionality of the process-based model and suggests that percolation in the study area may be modelled much simpler than using Hydrus1D. The CART model shows that soil properties play a negligible role for percolation under wet climatic conditions. However, they become more important if the conditions turn drier. The LR method does not yield satisfactory predictions for the spatially distributed percolation however the spatially aggregated percolation is well approximated. This may indicate that the soils behave simpler (i.e. more linear) when percolation dynamics are upscaled.     

The use of quantitative bacteriologic assessment of bone

A new sub-class of binding protein-dependent transporter with specificity for a broad range of polar amino acids has been identified by sequence comparison, in Rhizobium leguminosarum, Rhodobacter capsulatus, Escherichia coli and Pseudomonas fluorescens. Southern blotting and PCR analysis has shown that transporters from this new sub-class are widely distributed in Gram-negative bacteria, including, in addition to the above, Citrobacter freundii, Erwinia carotovorum and Rhizobium meliloti. ABC transporters of polar amino acids can be divided into two groups: those with narrow solute specificity and the newly identified sub-class with broad solute specificity. The binding and inner membrane proteins from transporters with a broad solute specificity are larger by approximately 30% than those with a narrow solute specificity. Multiple alignment of the inner membrane proteins from all sequenced polar amino acid transporters indicates there is an N-terminal conserved region that may be involved in solute specificity. A conserved arginine or lysine at residue 30 of this region is changed to glutamate in arginine transporters. Residue 53 also has a strong correlation with the charge on the transported solute, with basic amino acid transporters replacing an aliphatic amino acid at this position with a negatively charged amino acid. The general amino acid permease from R. leguminosarum, which will transport aliphatic as well as basic and acidic amino acids, juxtaposes two prolines at residues 52 and 53 of the N-terminal conserved region.     

Gambling in Norway: An obscure activity?

Suggests that it is difficult to get an exact account of gambling behavior in Norway. It appears that gambling behavior in Norway has increased 50% during the observed 5-yr period. Little is known about the prevalence of problematic gambling. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Fungicidal effect of 15 disinfectants against 25 fungal contaminants commonly found in bread and cheese manufacturing.

Resistance of 19 mold and 6 yeast species to 15 commercial disinfectants was investigated by using a suspension method in which the fungicidal effect and germination time were determined at 20 degrees C. Disinfectants containing 0.5% dodecyldiethylentriaminacetic acid, 10 g of chloramine-T per 1, 2.0% formaldehyde, 0.1% potassium hydroxide, 3.0% hydrogen peroxide, or 0.3% peracetic acid were ineffective as fungicides. The fungicidal effect of quaternary ammonium compounds and chlorine compounds showed great variability between species and among the six isolates of Penicillium roqueforti var. roqueforti tested. The isolates of P roqueforti var. carneum, P. discolor, Aspergillus versicolor, and Eurotium repens examined were resistant to different quaternary ammonium compounds. Conidia and vegetative cells were killed by alcohols, whereas ascospores were resistant. Resistance of ascospores to 70% ethanol increased with age. Both P. roqueforti var. roqueforti and E. repens showed great variability of resistance within isolates of each species.     

Room electromagnetics

In analogy with the established discipline of room acoustics, various aspects of diffuse wideband microwave propagation in a room are treated. It is shown that an equivalent to Sabine's equation for reverberation time in a room is valid for the completely diffused field, depending only on the volume, the surface area, and an effective absorption coefficient. An exponential decay of the power as a function of the delay is a consequence of the assumptions. Furthermore, the concept of a reverberation distance is also valid. This is the distance from a transmitting antenna where the received diffuse, randomly scattered power equals the direct line-of-sight received power, such that the diffuse power dominates for distances larger than the reverberation distance. A number of measurements in a large room support the theory with an effective absorption coefficient of 0.5. The power delay profiles around the room from a transmitter in the ceiling vary only in the first arriving part of the impulse, whereas the tail, being dominated by the diffuse field, has the same power level for a given delay and the same decay rate all over the room. It is also a consequence of the theory that the diffuse fields incident on an antenna are uniformly distributed in angle.     

Sites of alcohol and volatile anaesthetic action on GABA(A) and glycine receptors

Volatile anaesthetics have historically been considered to act in a nonspecific manner on the central nervous system. More recent studies, however, have revealed that the receptors for inhibitory neurotransmitters such as gamma-aminobutyric acid (GABA) and glycine are sensitive to clinically relevant concentrations of inhaled anaesthetics. The function of GABA(A) and glycine receptors is enhanced by a number of anaesthetics and alcohols, whereas activity of the related GABA rho1 receptor is reduced. We have used this difference in pharmacology to investigate the molecular basis for modulation of these receptors by anaesthetics and alcohols. By using chimaeric receptor constructs, we have identified a region of 45 amino-acid residues that is both necessary and sufficient for the enhancement of receptor function. Within this region, two specific amino-acid residues in transmembrane domains 2 and 3 are critical for allosteric modulation of both GABA(A) and glycine receptors by alcohols and two volatile anaesthetics. These observations support the idea that anaesthetics exert a specific effect on these ion-channel proteins, and allow for the future testing of specific hypotheses of the action of anaesthetics.     

The effect of cyclic stress on the physical properties of a poly(dimethylsiloxane) elastomer

The dynamic creep behavior of a filled poly(dimethylsiloxane) elastomer was studied under cyclic stress. The stress level was chosen such that the increase in the internal temperature was small and that microcracks were not observed. This work has demonstrated that cyclic stress in combination with high temperature accelerates the degradation of the elastomer. The results suggest that because of the applied force, breaks in the load-bearing chains of the network occur. These breaks, while relieving the mechanical stress, create highly reactive ionic fragments. It is believed that because of the subsequent reactions of the ionic fragments, changes in the specific gravity, storage modulus, effective crosslink density, and length of the sample (creep) are observed. The observed decrease in the storage modulus is thought to occur because of the reaction of the ionic fragments with moisture, which results in the formation of silanol chain ends that reduce the effective crosslink density. The results also show that contrary to the prediction of the Boltzmann's Superposition Principle, the rate of creep is greatly enhanced when the sample is subjected to a sinusoidally varying dynamic load as compared to a comparable static load. The polymer weight loss was found to be linear with time and strongly dependent on the level of applied dynamic and static force. In addition, the weight loss and rate of creep were also found to be strongly dependent upon temperature.