Development of Photoactivatable Allosteric Modulators for the Chemokine Receptor CXCR3

The CXCR3 receptor, a class A G protein‐coupled receptor (GPCR), is involved in the regulation and trafficking of various immune cells. CXCR3 antagonists have been proposed to be beneficial for the treatment of a wide range of disorders including but not limited to inflammatory and autoimmune diseases. The structure‐based design of CXCR3 ligands remains, however, hampered by a lack of structural information describing in detail the interactions between an allosteric ligand and the receptor. We designed and synthesized photoactivatable probes for the structural and functional characterization, using photoaffinity labeling followed by mass spectrometry, of the CXCR3 allosteric binding pocket of AMG 487 and RAMX3, two potent and selective CXCR3 negative allosteric modulators. Photoaffinity labeling is a common approach to elucidate binding modes of small‐molecule ligands of GPCRs through the aid of photoactivatable probes that convert to extremely reactive intermediates upon photolysis. The photolabile probe N‐[({1‐[3‐(4‐ethoxyphenyl)‐4‐oxo‐3,4‐dihydropyrido[2,3‐d]pyrimidin‐2‐yl]ethyl}‐2‐[4‐fluoro‐3‐(trifluoromethyl)phenyl]‐N‐{1‐[4‐(3‐(trifluoromethyl)‐3H‐diazirin‐3‐yl]benzyl}piperidin‐4‐yl)methyl]acetamide ( 10 ) showed significant labeling of the CXCR3 receptor (80 %) in a [³H]RAMX3 radioligand displacement assay. Compound 10 will serve as an important tool compound for the detailed investigation of the binding pocket of CXCR3 by mass spectrometry.     

Externalizing psychopathology and gain–loss feedback in a simulated gambling task: Dissociable components of brain response revealed by time-frequency analysis.

Externalizing is a broad construct that reflects propensity toward a variety of impulse control problems, including antisocial personality disorder and substance use disorders. Two event-related potential responses known to be reduced among individuals high in externalizing proneness are the P300, which reflects postperceptual processing of a stimulus, and the error-related negativity (ERN), which indexes performance monitoring based on endogenous representations. In the current study, the authors used a simulated gambling task to examine the relation between externalizing proneness and the feedback-related negativity (FRN), a brain response that indexes performance monitoring related to exogenous cues, which is thought to be highly related to the ERN. Time-frequency (TF) analysis was used to disentangle the FRN from the accompanying P300 response to feedback cues by parsing the overall feedback-locked potential into distinctive theta (4–7 Hz) and delta (     

Point mutations modifying the thrombin inhibition kinetics and antithrombotic activity in vivo of recombinant hirudin

The hirudin variant HV2 was modified by in vitro site-specificmutagenesis of HV2 cDNA to generate HV2(Asn-47 – Lys),HV2(Asn-47 – Arg) and HV2(Lys-35 – Thr, Asn-47 –Lys). Residues 35 and 47 are positioned respectively withinthe finger and prothrombin-like domains of hirudin, both ofwhich have been suggested as thrombin binding sites. The modifiedpolypeptides were synthesized in Saccharomyces cerevisiae usinga secretion vector and purified from culture supernatants. Byanalysis of the human -thrombin: hirudin inhibition reactionin steady-state conditions it was shown that the dissociationconstants for HV2(Lys-47) and HV2(Arg-47) were 5- to 14-foldlower than for unmodified HV2, whereas mutation of Lys-35 didnot significantly alter the inhibition kinetics. Furthermore,HV2(Lys-47), whose sequence is identical to a natural hirudinvariant, displayed enhanced antithrombotic activity in vivo,having a 100-fold lower ED₅₀ compared to HV2 in the rabbit Wesslervenous thrombosis model. These results support a role for theprothrombin-like domain in thrombin binding and, moreover, demonstratethat in vivo antithrombotic efficiency correlates with the dissociationconstant of the inhibition reaction.     

Quality of neurological care. Balancing cost control and ethics

As the quality of neurological care becomes a mutual objective of physicians, patients, and health planners, increased demands on cost savings will create conflicts that could threaten the ethical basis of medical practice. Physicians will see increasing ethical conflicts between their fiduciary duties to make treatment decisions in the best interest of their patients and their justice-based duties to conserve societal resources. These conflicts can be best mitigated if physicians maintain their orientation as patient advocates but practice cost-conscious clinical behaviors that consider the cost-effectiveness of tests and treatments and do not squander society's finite resources by ordering medical tests and treatments of zero or marginal utility. Health system planners should resolve their conflicting objectives of quality and cost control by rigorously defining and measuring quality through physician leadership and by implementing cost-control measures that enhance the quality of medical care. Managed care organizations voluntarily should forsake financially successful but blatantly unethical cost-saving schemes, such as gag clauses and end-of-year kickback payments to physicians, because these schemes diminish patients' trust in physicians and degrade the integrity of the patient-physician relationship. State and federal laws should prudently regulate these unethical cost-saving schemes to the same extent as they have for the harmful conflicts in fee-for-service medicine.     

Implications of Technological Changes in Vehicle Routing Interfaces for Planners' Constraint Processing

This study sought to assess the consequences of technological changes in vehicle routing interfaces for planners' constraint processing during route selection. An experiment featuring vehicle routing problems was designed to test interfaces reflecting technological changes, including automation leading to simplified interfaces and the display of multiple routes computed by algorithms. Twelve participants who had worked for a small transport company for 9 months were exposed to all these interfaces. Mental workload, performance, and decision‐making times were measured. Results revealed that automation decreases mental workload and decision times, attributable to the abridged (vs. unabridged) display of constraints on the interface. Results also showed that the perceptual (vs. analytical) display of routes greatly decreases decision times and enhances performances. Results are finally discussed in terms of information reliance, automation accuracy, and graphical forms tailored to meet genuine support needs. © 2012 Wiley Periodicals, Inc.     

Scheduler-oriented algorithms to improve human–machine cooperation in transportation scheduling support systems

A decision support system designed to enhance human–machine interaction in transportation scheduling is proposed. We aim to integrate human factors and ergonomics from the beginning of the design phase and to propose a system fitted with enough flexibility to be able to deal with the characteristics of a dynamic context such as transportation scheduling. In this interdisciplinary approach, a link is done between problem solving methods (operations research technics and data classification algorithms) and human–machine interaction (solving control modes). A set of scheduler-oriented algorithms favoring human–machine cooperation for problem solving is proposed. Some of these algorithms have been efficiently tested on instances of the literature. Finally, an original framework aiming to assist scheduler in constraint relaxation when the problem becomes infeasible is proposed and evaluated.     

Minimization of Sewage Network Overflow

We are interested in the optimal control of sewage networks. It is of high public interest to minimize the overflow of sewage onto the streets and to the natural environment that may occur during periods of heavy rain. The assumption of linear flow in a discrete time setting has proven to be adequate for the practical control of larger systems. However, the possibility of overflow introduces a nonlinear and nondifferentiable element to the formulation, by means of a maximum of linear terms. This particular challenge can be addressed by smoothing methods that result in a nonlinear program (NLP) or by logical constraints that result in a mixed integer linear program (MILP). We discuss both approaches and present a novel tailored branch-and-bound algorithm that outperforms competing methods from the literature for a set of realistic rain scenarios.     

Synthesis of silver nanoparticles by Bacillus subtilis T‐1 growing on agro‐industrial wastes and producing biosurfactant

In this study, culture supernatnats of Bacillus subtilis T‐1 growing on brewery effluents and molasses was used for silver nanoparticles (Ag‐NPs) synthesis. The biosurfactant production of B. subtilis T‐1 was confirmed by the detection of genes in the genome and by the identification of the product in the supernatants. The genes for synthesis of surfactin (sfp, srfAA) and iturin (ituC) were noted by PCR reactions. Also, in examined culture supernatants the presence of C₁₃, C₁₄ and C₁₅ surfactin homologues with the sodiated molecules [M + Na]⁺ at m /z 1030, 1044 and 1058 was confirmed using LC/MS/MS analysis. The formation of NPs in the culture supernatants was confirmed by UV–vis spectroscopy. The dynamic light scattering measurements and transmission electron microscopy images showed the nanometric sizes of the biosynthesised Ag‐NPs which ranged from several nm to several tens of nm depending on the used culture supernatant. Biological properties of Ag‐NPs were evaluated by binding of Ag‐NPs with DNA isolated from the Escherichia coli ATCC 25922 and B. subtilis ATCC 6633. Biogenic Ag‐NPs were actively bound to DNA in increased concentration which could be the one important mode of antibacterial action of the Ag‐NPs.Inspec keywords: silver, nanoparticles, nanofabrication, materials preparation, microorganisms, antibacterial activity, industrial waste, agrochemicals, surfactants, breweries, genomics, genetics, chromatography, mass spectroscopic chemical analysis, ultraviolet spectroscopy, visible spectroscopy, spectrochemical analysis, light scattering, transmission electron microscopy, DNA, bonds (chemical), biochemistry, molecular biophysics, nanobiotechnology, biological techniques, particle size, enzymesOther keywords: silver nanoparticle synthesis, Bacillus subtilis T‐1 growth, agro‐industrial waste, biosurfactant production, brewery effluent, molasses, Ag‐NP synthesis, B. subtilis T‐1, gene detection, genome, supernatant product identification, surfactin synthesis, sfp, srfAA, iturin synthesis, ituC, PCR reaction, C₁₃ surfactin homologue, C₁₄ surfactin homologue, C₁₅ surfactin homologue, sodiated molecules, LC‐MS‐MS analysis, UV‐vis spectroscopy, dynamic light scattering measurement, transmission electron microscopy image, Ag‐NP nanometric size range, Ag‐NP biosynthesis, used culture supernatant dependence, biological properties, DNA isolation, Escherichia coli ATCC 25922, B. subtilis ATCC 6633, biogenic Ag‐NP‐DNA binding, Ag‐NP antibacterial action, Ag     

Diversity,competition, extinction: the ecophysics of language change

As indicated early by Charles Darwin, languages behave and change very much like living species. They display high diversity, differentiate in space and time, emerge and disappear. A large body of literature has explored the role of information exchanges and communicative constraints in groups of agents under selective scenarios. These models have been very helpful in providing a rationale on how complex forms of communication emerge under evolutionary pressures. However, other patterns of large-scale organization can be described using mathematical methods ignoring communicative traits. These approaches consider shorter time scales and have been developed by exploiting both theoretical ecology and statistical physics methods. The models are reviewed here and include extinction, invasion, origination, spatial organization, coexistence and diversity as key concepts and are very simple in their defining rules. Such simplicity is used in order to catch the most fundamental laws of organization and those universal ingredients responsible for qualitative traits. The similarities between observed and predicted patterns indicate that an ecological theory of language is emerging, supporting (on a quantitative basis) its ecological nature, although key differences are also present. Here, we critically review some recent advances and outline their implications and limitations as well as highlight problems for future research.     

Design Proposal to Avoid Peeling Failure in FRP-Strengthened Reinforced Concrete Beams

According to the available experimental work, the most common failure in existing structures strengthened by plate bonding is the laminate peeling off. In the last few years, an important effort in the development of mathematical models to avoid premature peeling failures has been made. However, a suitable and reliable design method to predict debonding due to the shear flow between crack discontinuities or at the laminate end is still not available. This paper describes a new design procedure for structures strengthened by plate bonding to avoid peeling failure at any location. After calculating the laminate area required for flexural strengthening, a two-step procedure to prevent peeling failure is proposed. The first step, to avoid peeling failure along the span, is based on a shear-bending interaction diagram associated with the theoretical maximum transferred force between laminate and support along the crack spacing before laminate debonding. This interaction diagram can be obtained through the application of nonlinear fracture mechanics. The second step consists of checking for peeling failure at the laminate end. The bonded length between the laminate end and the nearest crack should be enough to transfer the laminate tensile force acting on this crack. The proposed method has been verified with available experimental results assembled in a database. A good agreement between the experimental and predicted failure load has been obtained. Finally, an application example is presented to show the applicability of the method.