Thioether Analogues of Disulfide‐Bridged Cyclic Peptides Targeting Death Receptor 5: Conformational Analysis,Dimerisation and Consequences for Receptor Activation

Cyclic peptides containing redox‐stable thioether bridges might provide a useful alternative to disulfide‐bridged bioactive peptides. We report the effect of replacing the disulfide bridge with a lanthionine linkage in a 16‐mer cyclic peptide that binds to death receptor 5 (DR5, TRAIL‐R2). Upon covalent oligomerisation, the disulfide‐bridged peptide has previously shown similar behaviour to that of TNF‐related apoptosis inducing ligand (TRAIL), by selectively triggering the DR5 cell death pathway. The structural and biological properties of the DR5‐binding peptide and its desulfurised analogue were compared. Surface plasmon resonance (SPR) data suggest that these peptides bind DR5 with comparable affinities. The same holds true for dimeric versions of these peptides: the thioether is able to induce DR5‐mediated apoptosis of BJAB lymphoma and tumorigenic BJELR cells, albeit to a slightly lower extent compared to its disulfide homologue. NMR analysis revealed subtle variation in the conformations of the two peptides and suggests that the thioether peptide is slightly less folded than its disulfide homologue. These observations could account for the different capability of the two dimers to cluster DR5 receptors on the cell surface and to trigger apoptosis. Nevertheless, our results suggest that the thioether peptide is a potential candidate for evaluation in animal models.     

Influence of High-Pressure Coolant Assistance on the Machinability of the Titanium Alloy Ti555–3

The origin of this article is the quantification of productivity gains and the improvement in surface integrity seen for a recent titanium alloy that is seeing increasing use in the aeronautical industry. The Ti555–3 titanium alloy, which is starting to find greater application in the aeronautical field, exhibits certain difficulties in terms of machining. High Pressure Coolant (HPC) assisted turning consists of projecting a high pressure coolant jet between the chip and the tool. Comparisons are made between assisted turning using variable jet pressure and conventional turning (dry and classical lubrication). It is shown that it is possible to improve productivity by using HPC-assisted machining. The results highlight good chip fragmentation and a great improvement of tool life with HPC assistance. Surface integrity is also shown to be improved, through surface roughness parameters that decrease, and surface residual stresses that become more compressive. These effects have been attributed to the thermo-mechanical action of the coolant jet resulting in lower cutting forces, lower coefficient of friction and lower temperature in the cutting zone.     

A long-term view of nutrient transfers through the Seine river continuum

A model (the Riverstrahler model) is used to describe nutrient transfer and transformation at the scale of the whole drainage network of the Seine based on information concerning the basic mechanisms governing N, P and Si inputs to the drainage network and in-stream transformation and retention. It was used to calculate the budget of these nutrients through the whole river continuum from land to sea. With the help of historical documents, the constraints used as forcing function in this model were reconstructed to express the changing conditions of land-use and urban population over the last five hundred years. The corresponding scenarios were run for different hydrological regimes including dry, mean and wet conditions. The results were validated on the long-term series of nutrient measurements spanning more than a century available at some stations on the Seine, upstream and downstream of the city of Paris. The model was also used to explore past and future trends in nutrient loading, retention and delivery to the coastal zone, in response to human management of the terrestrial watershed. Beside the initial pristine state, used as an idealized reference state (with N, P and Si delivered fluxes of about 45-110 kg N km(-2) yr(-1), 2-5 kg P km(-2) yr(-1), 510-1325 kg Si km(-2) yr(-1)), four periods were distinguished. The first one is that of the traditional cottage economy which prevailed, with quite a constant total population, until the end of the 18th century. N, P and Si fluxes were about 235-750 kg N km(-2) yr(-1), 15-60 kg P km(-2) yr(-1) and 425-1280 kg Si km(-2) yr(-1), depending on hydrological conditions. The second period, from the beginning of the 19th century to about the 1950's, corresponded to rapid increase in the total and urban population with a corresponding increase of point sources of N and P. From 1950 onwards, modern farming practices resulted in a dramatic increase in diffuse sources of nitrogen and to a lesser extent phosphorus: riverine N and P export reached 1320-2800 kg N km(-2) yr(-1), and 310-340 kg P km(-2) yr(-1): silica export remained fairly constant at around 410-1260 kg Si km(-2) yr(-1) depending on the hydrological conditions. In the 1990's, the fourth period is represented by a stabilized population and improved wastewater treatment, when the export of phosphorus is reduced to values as low as 40-60 kg P km(-2) yr(-1), but without as effective a reduction of nitrogen export. This represents an unprecedented situation for the marine coastal system, i.e. a shift from nitrogen to phosphorus limitation, as nitrogen is still delivered far in excess of the amount of silica available for diatom blooms.     

Evaluation of enrichment-free PCR-based detection on the rfbE gene of Escherichia coli O157--application to municipal wastewater

Escherichia coli O157 strains have emerged as important human enteric pathogens. Strains that express the O-antigen 157 are commonly associated with severe clinical manifestations, including bloody diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome. E. coli O157 strains may be transmitted in a variety of ways, including food, water and person-to-person or animal-to-person contact. Fecal contamination is one source of environmental contamination and is responsible for the presence of these pathogens in the environment. We used a specific and sensitive PCR assay based on the rfbE gene to detect low levels of these pathogens in wastewater. The set of primers used was designed to amplify an intragenic segment of the rfbE gene. The amplification assay detected 200 CFU of E. coli O157 in pure water. The prevalence of E. coli O157 in the effluents of 44 wastewater treatment plants was determined (7%).     

Impact of microparticles on UV disinfection of indigenous aerobic spores

Numerous studies have shown that the efficacy of ultraviolet (UV) disinfection can be hindered by the presence of particles that can shield microorganisms. The main objective of this study was to determine to what extent natural particulate matter can shield indigenous spores of aerobic spore-forming bacteria (ASFB) from UV rays. The extent of the protective shielding was assessed by comparing the inactivation rates in three water fractions (untreated, dispersed and filtered on an 8 microm membrane) using a collimated beam apparatus with a low-pressure lamp emitting at 254 nm. Levels of inactivation were then related to the distribution and abundance of particles as measured by microflow imaging. Disinfection assays were completed on two source waters of different quality and particle content. A protocol was developed to break down particles and disperse aggregates (addition of 100mg/L of Zwittergent 3-12 and blending at 8000 rpm for 4 min). Particle size distribution (PSD) analysis confirmed a statistically significant decrease in the number of particles for diameter ranges above 5 microm following the dispersion protocol and 8 microm filtration. The fluence required to reach 1-log inactivation of ASFB spores was independent of particle concentration, while that required to reach 2-log inactivation or more was correlated with the concentration of particles larger than 8 microm (R(2)>0.61). Results suggest that natural particulate matter can protect indigenous organisms from UV radiation in waters with elevated particle content, while source water with low particle counts may not be subject to this interference.     

Parameter estimation and prediction uncertainties for multi-response kinetic models with uncertain inputs

Error-in-variables model (EVM) methods are used for parameter estimation when independent variables are uncertain. During EVM parameter estimation, output measurement variances are required as weighting factors in the objective function. These variances can be estimated based on data from replicate experiments. However, conducting replicates is complicated when independent variables are uncertain. Instead, pseudo-replicate runs may be performed where the target values of inputs for repeated runs are the same, but the true input values may be different. Here, we propose a method to estimate output-measurement variances for use in multivariate EVM estimation problems, based on pseudo-replicate data. We also propose a bootstrap technique for quantifying uncertainties in resulting parameter estimates and model predictions. The methods are illustrated using a case study involving n-hexane hydroisomerization in a well-mixed reactor. Case-study results reveal that assumptions about input uncertainties can have important influences on parameter estimates, model predictions and their confidence intervals.     

Additional vectors for PCR-based gene tagging in Saccharomyces cerevisiae and Schizosaccharomyces pombe using nourseothricin resistance

The one-step PCR-mediated technique used for modification of chromosomal loci is a powerful tool for functional analysis in yeast. Both Saccharomyces cerevisiae and Schizosaccharomyces pombe are amenable to this technique. However, the scarce availability of selectable markers for Sz. pombe hampers the easy use of this technique in this species. Here, we describe the construction of new vectors deriving from the pFA6a family, which are suitable for tagging in both yeasts owing to the presence of a nourseothricin-resistance cassette. These plasmids allow various gene manipulations at chromosomal loci, viz. N- and C-terminal tagging with 3HA (haemagglutinin) or 13Myc epitopes, GST (glutathione S-transferase), 4TAP (tandem affinity purification) and several GFP (green fluorescent protein) isoforms. For N-terminal modifications, the use of different promoters allows constitutive (PADH1) or regulatable (PGAL1) promoters for S. cerevisiae and derivatives of Pnmt1 for Sz. pombe expression.     

Encoding and selective activation of "metabolic memories" in the rat.

Experiment 1 used Pavlovian conditioning procedures to show that rats formed distinct memorial representations of 2 (peanut oil and sucrose pellets) unconditioned stimuli (USs) that could be activated by 2 different conditioned stimuli (CSs). After training in Experiment 2, rats injected with the lipid antimetabolite Na-2-mercaptoacetate (MA) responded more to the CS for oil than to the CS for sucrose. This pattern was not shown by rats that received isotonic saline or systemic 2-deoxy-d-glucose (a glucose antimetabolite). By contrast, intracerebroventricular infusion of the glucose antimetabolite 5-thioglucose selectively promoted responding to the CS for sucrose (Experiment 4). Thus, lipoprivic and glucoprivic treatments selectively promoted the activation of the memories of fat and carbohydrate USs, respectively. In Experiment 3, the capacity of MA to augment responding to a CS for oil was abolished for rats that received subdiaphragmatic vagal deafferentation. This indicates that the capacity of lipoprivic signals to selectively activate the representations of fat USs may depend on vagal afferent fibers. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Toxic effects of hexavalent chromium on brook trout (Salvelinus fontinalis) and rainbow trout (Salmo gairdneri)

Exposing brook trout to various concentrations of chromium [Cr(VI)] for up to 22 months (including reproduction) significantly increased alevin mortality at 0.35 mg Cr l⁻¹ and retarded growth of young brook trout at the lowest concentration tested (0.01 mg Cr l⁻¹). Eight month exposures of rainbow trout significantly increased alevin mortality at 0.34 mg Cr l⁻¹ and also retarded growth at the lowest concentration tested (0.10 mg Cr l⁻¹). Exposures of brook trout lasting 22 months showed, however, that growth was only temporarily affected, and therefore, it was not used as an end point to measure the affects of chromium on either species. Reproduction, and embryo hatchability of brook trout were unaffected at Cr(VI) concentrations that affected survival of newly hatched alevins. The maximum acceptable toxicant concentration (MATC) for brook and rainbow trout exposed to Cr(VI) in water with a hardness of 45 mg l⁻¹ (as CaCO₃) and a pH range of 7–8 lies between 0.20 and 0.35 mg Cr l⁻¹. The 96-h lc₅₀ for brook and rainbow trout was 59 and 69 mg Cr l⁻¹, respectively: therefore, the application factor (MATC/96-h lc₅₀) for both species lies between 0.003 and 0.006.     

WIRELESS BODY-AREA NETWORKS

The last decade has witnessed the convergence of three giant worlds:electronics,computer science and telecommunications.The next decade should follow this convergence in most of our activities with the generalization of sensor networks.In particular with the progress in medicine,people live longer and the aging of population will push the development of wireless person-