Integrative Study of the Structural and Dynamical Properties of a KirBac3.1 Mutant: Functional Implication of a Highly Conserved Tryptophan in the Transmembrane Domain

ATP-sensitive potassium (K-ATP) channels are ubiquitously expressed on the plasma membrane of cells in several organs, including the heart, pancreas, and brain, and they govern a wide range of physiological processes. In pancreatic β-cells, K-ATP channels composed of Kir6.2 and SUR1 play a key role in coupling blood glucose and insulin secretion. A tryptophan residue located at the cytosolic end of the transmembrane helix is highly conserved in eukaryote and prokaryote Kir channels. Any mutation on this amino acid causes a gain of function and neonatal diabetes mellitus. In this study, we have investigated the effect of mutation on this highly conserved residue on a KirBac channel (prokaryotic homolog of mammalian Kir6.2). We provide the crystal structure of the mutant KirBac3.1 W46R (equivalent to W68R in Kir6.2) and its conformational flexibility properties using HDX-MS. In addition, the detailed dynamical view of the mutant during the gating was investigated using the in silico method. Finally, functional assays have been performed. A comparison of important structural determinants for the gating mechanism between the wild type KirBac and the mutant W46R suggests interesting structural and dynamical clues and a mechanism of action of the mutation that leads to the gain of function.     

A fragment-based approach to understanding inhibition of 1-deoxy-D-xylulose-5-phosphate reductoisomerase

The inhibition of 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) by fosmidomycin was studied by using a kinetic assay based on the consumption of NADPH and synthetic substrate. Fosmidomycin is a slow tight-binding inhibitor of DXR that shows strong negative cooperativity (absolute value(h) = 0.3) in binding. Cooperativity is displayed during the initial (weak, K0.5 = 10 microM) binding event and does not change as the binding tightens to the equilibrium value of 0.9 nM over a period of seconds to minutes. A series of fosmidomycin fragments was examined, but all showed much weaker inhibition, in the mM range. A series of cyclic fosmidomycin analogues was also synthesised and tested, but these showed high-microM binding at best. None of the synthetic compounds showed time-dependent inhibition. We concluded that the slow tight-binding behaviour, and perhaps also cooperativity, are mediated by significant reorganisation of the active site upon fosmidomycin binding. This makes the rational design of new inhibitors of DXR difficult at best.     

Searching information with a natural language dialogue system: a comparison of spoken vs. written modalities

This paper examines the effects of spoken vs. written dialogue modalities on the effectiveness of information search with a computerized retrieval system. Forty-eight adults familiar with the use of computers were asked to carry out six information retrieval tasks, engaging with the system using either spoken or written communication. The written modality was more efficient with regard to the number of dialogue turns, length of interaction with the system and mental workload. Even though the turns lasted longer in the written mode, they appeared to yield less mental workload. Moreover, spoken and written dialogues did not differ as regards the use of pronouns and articles. The implications for the development of natural-language dialogue systems are discussed.     

Reduction of aqueous chromate by Fe(II)/Fe(III) carbonate green rust: kinetic and mechanistic studies

This work describes the heterogeneous reaction between FeII in carbonate green rust and aqueous chromate, in NaHCO3 solutions at 25 degrees C, and at pH values of 9.3-9.6. Evidence for reduction of CrVI to CrIII and concomitant solid-state oxidation of lattice FeII to FeIII was found from FeII titration and from structural analysis of the solids using FTIR, XRD, SEM, and XPS methods. Results indicate the formation of ferric oxyhydroxycarbonate and the concomitant precipitation of CrIII monolayers at the surface of the iron compound that induce passivation effects and progressive rate limitations. The number of CrIII monolayers formed at the completion of the reaction depends on [FeII]t=0, the molar concentration of FeII(solid) at t=0; on [n(o)]t=0, the molar concentration of reaction sites present at the surface of the solid phase at t=0; and on [CrVI]t=0, the molar concentration of CrVI at t=0. Kinetic data were modeled using a model based on the formation of successive CrIII monolayers, -(d[CrVI]/dt) = sigma(1)j k(i)[S] [CrVI]([n(i - 1)] - [n(i)]) with k(i)[S] (in s(-1) L mol(-1)), the rate coefficient of formation of CrIII monolayer i, and [n(i)] and [n(i - 1)], the molar concentration of CrIII precipitated in monolayer i and monolayer i - 1, respectively. Good matching curves were obtained with kinetic coefficients: k(1)[S] = 5-8 x 10(-4), k(2)[S] = 0.5-3 x 10(-5), and k(3)[S] about 1.7 x 10(-6) s(-1) m(-2) L. The CrVI removal efficiency progressively decreases along with the accumulation of CrIII monolayers at the surface of carbonate green rust particles. In the case of thick green rust particles resulting from the corrosion of iron in permeable reactive barriers, the quantity of FeII readily accessible for efficient CrVI removal should be rather low.     

Is Dimethyldecanal a Common Aggregation Pheromone of Tribolium Flour Beetles?

Flour beetles are cosmopolitan and common pests in grain stores and flour mills. Their ability to exploit a wide variety of stored products has contributed to their status as major pests of stored food. Although it was previously reported that the same aggregation pheromone, 4,8-dimethyldecanal (DMD), is shared by three flour beetles species (Tribolium castaneum, T. confusum, and T. freemani), the volatiles released by the other Tribolium species associated with stored products have not yet been examined. In the present study, the volatiles produced by males and females of eight Tribolium species were examined by solid phase microextraction (SPME). SPME samples were analyzed by gas chromatography coupled to mass spectrometry (GC-MS). Experiments were conducted to identify volatiles emitted by the adults of different Tribolium species and to determine whether DMD is a common aggregation pheromone. We observed that DMD is not a common pheromone of the eight species tested, but is common to T. castaneum, T. confusum, T. freemani, and T. madens. Two other volatiles were detected, 1-pentadecene, which is shown here to be a common semiochemical of flour beetles, and 1,6-pentadecadiene, which was detected in five species (T. audax, T. brevicornis, T. destructor, T. freemani, and T. madens).     

Engine calibration: multi-objective constrained optimization of engine maps

We present two new approaches to address the optimization problem associated with engine calibration. In this area, the tuning parameters are traditionally determined in a local way, i.e., at each engine operating point, via a single-objective minimization problem. To overcome these restrictions, the first method we propose is able to cope with several objective functions simultaneously in the local formulation. The second method we put forward relies on a global formulation, which allows the whole driving cycle to be taken into account while remaining single-objective. At the practical level, the two methods are implemented by combining various existing techniques such as the LoLiMoT (Local Linear Model Tree) parameterization and the MO-CMA-ES (Multi-Objective Covariance Matrix Adaptation Evolution Strategy) algorithm. A better compromise appears to be achieved on real case applications.     

Long-term Uncertainty of Hydropower Revenue Due to Climate Change and Electricity Prices

Hydropower represents the world’s largest renewable energy source. As a flexible technology, it enhances reliability and security of the electricity system. However, climate change and market liberalization may hinder investment due to the evolution of water runoffs and electricity prices. Both alter expected revenue and bring uncertainty. It increases risk and deters investment. Our research assesses how climate change and market fluctuation affect annual revenue. But this paper focuses on the uncertainty, rather than on forecasting. This transdisciplinary topic is investigated by means of a mixed method, i.e. both quantitative and qualitative. The quantitative approach uses established models in natural sciences and economics. The uncertainty is accounted for by applying various scenarios and various datasets coming from different models. Based on those results, uncertainty is discussed through an analysis discerning three dimensions of uncertainty. Uncertainty analysis requires the assessment of a large panel of models and data sets. It is therefore rarely carried out. The originality of the paper also lies on the combination of quantitative established models with a qualitative analysis. The results surprisingly show that the greenhouse gas scenarios may in fact represent a low source of uncertainty, unlike electricity prices. Like forecasting, the main uncertainties are actually case study related and depend on the investigated variables. It is also shown that the nature of uncertainty evolves. Runoff uncertainty goes from variability, i.e. inherent randomness, to epistemic, i.e. limitation of science. The reverse situation occurs with the electricity price. The implications for scientists and policy makers are discussed.     

An enhanced Lemaitre model formulation for materials processing damage computation

The Lemaitre damage model is now widely used to deal with coupled damage analyses for various mechanical applications. In this article, different extensions of the model are presented and discussed to deal with complex multiaxial configurations—such as multi-stages bulk forming processes. A specific treatment is done to account for compressive damage growth, and a stress triaxiality cut-off value is considered to avoid any damage evolution below a critical negative triaxiality. The damage potential is also modified to deal with highly ductile materials, and the plastic strain is split into a negative part and a positive part to differentiate damage growth for compressive states of stress and for tensile states of stress. Finally, an anisotropic damage approach based on the comparison between grain flow orientation and principal loading directions is defined. A combination of these extensions is achieved within a single Lemaitre formulation. Application on different examples show the robustness and accuracy of the model defined in this paper.     

Hydraulic Modeling of a Mixed Water Level Control Hydromechanical Gate

This article describes the hydraulic behavior of a mixed water level control hydromechanical gate present in several irrigation canals. The automatic gate is termed “mixed” because it can hold either the upstream water level or the downstream water level constant according to the flow conditions. Such a complex behavior is obtained through a series of side tanks linked by orifices and weirs. No energy supply is needed in this regulation process. The mixed flow gate is analyzed and a mathematical model for its function is proposed, assuming the system is at equilibrium. The goal of the modeling was to better understand the mixed gate function and to help adjust their characteristics in the field or in a design process. The proposed model is analyzed and evaluated using real data collected on a canal in the south of France. The results show the ability of the model to reproduce the function of this complex hydromechanical system. The mathematical model is also implemented in software dedicated to hydraulic modeling of irrigation canals, which can be used to design and evaluate management strategies.