Aequorin-based functional assays for G-protein-coupled receptors,ion channels,and tyrosine kinase receptors

Aequorin is a photoprotein originating from jellyfish, whose luminescent activity is dependent on the concentration of calcium ions. Due to the high sensitivity and low background linked to luminescent assays, as well as to its absence of toxicity and its large linear dynamic range, aequorin has been used as an intracellular calcium indicator since its discovery in the early 1960s. The first applications of aequorin involved its microinjection in cells. The cloning of its gene in 1985 opened the way to the stable expression of aequorin in cell lines or even entire organisms. Here we present the validation of aequorin as a functional assay for the screening of G-protein-coupled receptors, ion channels, and tyrosine kinase receptors, as well as for their pharmacological characterization in agonist and antagonist detection assays. We optimized our cell suspension-based assay and determined that the most sensitive assay was performed at room temperature, with mitochondrially expressed aequorin and using coelenterazine derivative h for reconstitution of aequorin. The robustness of the assay and the current availability of luminometers with integrated injectors allow aequorin to fit perfectly with high throughput functional assays requirements.     

Population computation of vectorial transformations

Many neurons of the central nervous system are broadly tuned to some sensory or motor variables. This property allows one to assign to each neuron a preferred attribute (PA). The width of tuning curves and the distribution of PAs in a population of neurons tuned to a given variable define the collective behavior of the population. In this article, we study the relationship of the nature of the tuning curves, the distribution of PAs, and computational properties of linear neuronal populations. We show that noise-resistant distributed linear algebraic processing and learning can be implemented by a population of cosine tuned neurons assuming a nonuniform but regular distribution of PAs. We extend these results analytically to the noncosine tuning and uniform distribution case and show with a numerical simulation that the results remain valid for a nonuniform regular distribution of PAs for broad noncosine tuning curves. These observations provide a theoretical basis for modeling general nonlinear sensorimotor transformations as sets of local linearized representations.     

A Unifying and Rigorous Shape from Shading Method Adapted to Realistic Data and Applications

We propose a new method for the Lambertian Shape From Shading (SFS) problem based on the notion of Crandall-Lions viscosity solution. This method has the advantage of requiring the knowledge of the solution (the surface to be reconstructed) only on some part of the boundary and/or of the singular set (the set of the points at maximal intensity). Moreover it unifies in an unique mathematical formulation the works of Rouy et al. [34, 50], Falcone et al. [21], Prados et al. [46, 48, 49], based on the notion of viscosity solutions and the work of Dupuis and Oliensis [17] dealing with classical solutions and value functions. Also, it allows to generalize their results to the “perspective SFS” problem recently simultaneously introduced in [13,46,55]. While the theoretical part has been developed in [44], in this paper we give some stability results and we describe numerical schemes for the SFS based on this method. We construct provably convergent and robust algorithms. Finally, we apply our SFS method to real images and we suggest some real-life applications.     

Large-Eddy Simulation of the Lid-Driven Cubic Cavity Flow by the Spectral Element Method

This paper presents the large-eddy simulation of the lid-driven cubic cavity flow by the spectral element method (SEM) using the dynamic model. Two spectral filtering techniques suitable for these simulations have been implemented. Numerical results for Reynolds number Re=12,000 are showing very good agreement with other experimental and DNS results found in the literature.     

Uniform controllability of semidiscrete approximations of parabolic control systems

Controlling an approximation model of a controllable infinite dimensional linear control system does not necessarily yield a good approximation of the control needed for the continuous model. In the present paper, under the main assumptions that the discretized semigroup is uniformly analytic, and that the control operator is mildly unbounded, we prove that the semidiscrete approximation models are uniformly controllable. Moreover, we provide a computationally efficient way to compute the approximation controls. An example of application is implemented for the one- and two-dimensional heat equation with Neumann boundary control.     

Sex- and site-dependent variations in the thickness and mechanical properties of human skin in vivo

A new method for the in vivo characterization of the physical properties of skin is presented. This comprises an ultrasound device to measure the vertical displacement of the surface of the skin, as well as its thickness and that of the hypodermis under suction. In combination with this, a mathematical model is used to calculate the following skin parameters: Young's modulus, the initial stress and an index of non-elasticity. These parameters were evaluated from the volar forearm and the forehead of 30 male and 30 females, of similar ages (28 +/- 6-years-old). The sensitivity of the testing procedure, allowing the characterization of the mechanical parameters of the skin, easily differentiated these two sites, and in some cases, differences between women and men were demonstrated. The main results showed for both sexes that the thickness (P = 0.0001), Young's modulus (P = 0.0001), and the index of non-elasticity (P = 0.0001) were greater for the forehead than for the ventral forearm, but that the initial stress was lower (P = 0.0001). The results show that the skin is thicker, stiffer and less tense and elastic on the forehead than on the ventral forearm, suggesting structural differences between these two sites (collagen fibre network, elastic fibres, epidermis, stratum corneum, microvascularization, actinic damage, presence of sebaceous glands, etc.). It is hoped that this device will be useful for the evaluation of certain skin disorders (scleroderma, Ehlers-Danlos syndrome, cutis laxa, oedema, etc.) and their therapy, as well as being a useful tool in skin ageing and cosmetic product assessment.     

The Addiction-Stroop test: Theoretical considerations and procedural recommendations.

Decisions about using addictive substances are influenced by distractions by addiction-related stimuli, of which the user might be unaware. The addiction-Stroop task is a paradigm used to assess this distraction. The empirical evidence for the addiction-Stroop effect is critically reviewed, and meta-analyses of alcohol-related and smoking-related studies are presented. Studies finding the strongest effects were those in which participants had strong current concerns about an addictive substance or such concerns were highlighted through experimental manipulations, especially those depriving participants of the substance. Theories to account for addiction-related attentional bias are discussed, of which the motivational theory of current concerns appears to provide the most complete account of the phenomenon. Recommendations are made for maximizing the precision of the addiction-Stroop test in future research. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Covalent modifications of aminophospholipids by 4-hydroxynonenal

Lipid oxidation is implicated in a wide range of pathophysiological disorders, which leads to reactive compounds such as aldehydes. Among them 4-hydroxynonenal (4-HNE) reacts strongly with the NH2 groups of amino acids and forms mainly Michael adducts and minor Schiff-base adducts. Such reactions occur also with compounds containing thiol groups. No data are available describing 4-HNE interactions with amino-phospholipids. To investigate such a possibility, 4-HNE was incubated with either phosphatidylethanolamine (PE) or phosphatidylserine (PS) in an aqueous-organic biphasic system and the resulting products were identified by liquid chromatography-mass spectrometry (LC-MS). Our study points out the potential capacity of 4-HNE to react with phospholipids containing amino groups and particularly PE. The main resulting compounds found were a Michael adduct plus a minor Schiff base adduct, which was partly cyclized as a pyrrole derivative via a loss of water. Its stabilization as a pyrrole derivative allows to differentiate 4-HNE from the other aldehydes generated via lipid oxidation (e.g., malondialdehyde, 2-nonenal) that lack the 4-hydroxyl group. Their formation seems not to be affected when the pH varies from 6.5 to 8.5. Surprisingly, PS reacted poorly producing only a small amount of Michael adduct, the Schiff-base adduct being nondetectable. We conclude that such adducts, if they are formed in cell membranes, could alter the phospholipase-dependent cell signaling.     

Elastic wave velocities in various alloy strips

The zeroth order plate mode shear wave velocity has been measured in thin strip specimens of Oriented Electrical Steel, Elinvar-Extra, and RMI 464 titanium alloy as a function of the angle between the propagation direction and the rolling direction. Also, the plane wave shear and longitudinal velocities have been measured along the normal to the rolling plane. The results in the Oriented Electrical Steel agree with the known (110)[001] texture. In the Elinvar-Extra there is a texture indistinguishable elastically from (100)[Oil], or partial (100)[011] superimposed on a random background. Different annealing temperatures following cold-rolling yield different degrees of anisotropy and different dependences (1/G)(dG/dT) of the shear modulus upon ambient temperature. At an annealing temperature of about 850‡C, (1/G)(dG/dT) evaluated near room temperature changes from positive to negative, and anisotropy becomes minimum. In the RMI 464 titanium alloy, the shear velocity anisotropy was only 1 pct in the rolling plane. Formerly with Bell Telephone Laboratories, Allen town, Pa.