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.     

The location of a resistive shunt harmonic impedance along a distribution feeder and its influence on harmonic propagation

Although complete compensation is only possible using true active filters, resistive shunt harmonic impedances (SHI) have been shown to provide a considerable reduction of the harmonic propagation. In this paper, the harmonic mitigation potential of a resistive SHI is discussed concerning its influence on the voltage distortion profile along a typical distribution feeder, with the location of the SHI along the feeder as a parameter. Although the distortion values are dependent on the SHI location, it is shown that the end of the feeder is generally a good location to install the SHI, especially when the power system parameters can vary or are unknown. Calculations are performed on a typical radial distribution feeder. Both the power factor correction capacitors and the non-linear loads are concentrated in single nodes. The linear loads are disconnected to obtain the worst case for the voltage distortion. Also some measurements on a scale model of a typical distribution feeder are done and the experimental results confirm the results obtained from simulations.     

Effect of pindolol on hormone secretion and body temperature: partial agonist effects

Pindolol has been shown to be a partial agonist at 5-HT1A receptors in preclinical studies. It has also been reported to inhibit the effects of other 5-HT1A partial agonists such as ipsapirone and buspirone on hormone secretion and body temperature in man, indicating its antagonist action at 5-HT1A receptors in man. To determine if pindolol has 5-HT1A agonist as well as antagonist effects in man, pindolol, 30 mg, p.o. and placebo, were given single blind in random order to 23 normal men with indwelling venous catheters and its effects on hormone secretion and body temperature noted. Pindolol significantly increased basal plasma cortisol concentrations, whereas it decreased plasma prolactin (PRL) concentrations and body temperature. The increase in plasma cortisol due to pindolol suggests a 5-HT1A agonist action and is consistent with a 5-HT1A partial agonist mechanism in man whereas the PRL effects are consistent with an antagonist action at 5-HT1A receptors. The effects of pindolol on plasma cortisol concentration and body temperature were significantly negatively correlated. Furthermore, these results indicate significant differences in the 5-HT1A-dependent regulation of PRL and the hypothalamo-pituitary-adrenal (HPA) axis and body temperature, and suggest that human basal PRL secretion is tonically stimulated by 5-HT1A mechanism whereas the HPA axis and body temperature are not. Since rodent studies suggest differences in 5-HT1A receptor sensitivity between males and females, the results reported here need to be replicated in females. These differences in the effect of pindolol are discussed in terms of receptor reserve theory.     

Spreading resistance correction formula more suited for the Gauss-Laguerre quadrature

A modified formula is derived for the calculation of the correction factor in spreading resistance measurements. Due to its better convergence this formulation yields more accurate results when the Gauss-Laguerre quadrature formulae are used for the integration. In this way the obtained accuracy is found to be always better than 0.15% compared to 1.3% when the classic formulation is used. A very high accuracy for the correction factor is shown to be very important since an error of 2% in the correction factors may result in a 16% error in the derived resistivities.The CPU time when using the new formula is always smaller than or equal to the time used for the classic formulation and is typically about 15 seconds on a IBM 3033 computer for the calculation of 300 points.     

Dynamical diseases of brain systems: different routes to epileptic seizures

In this overview, we consider epilepsies as dynamical diseases of brain systems since they are manifestations of the property of neuronal networks to display multistable dynamics. To illustrate this concept we may assume that at least two states of the epileptic brain are possible: the interictal state characterized by a normal, apparently random, steady-state electroencephalography (EEG) ongoing activity, and the ictal state, that is characterized by paroxysmal occurrence of synchronous oscillations and is generally called, in neurology, a seizure. The transition between these two states can either occur: 1) as a continuous sequence of phases, like in some cases of mesial temporal lobe epilepsy (MTLE); or 2) as a sudden leap, like in most cases of absence seizures. In the mathematical terminology of nonlinear systems, we can say that in the first case the system's attractor gradually deforms from an interictal to an ictal attractor. The causes for such a deformation can be either endogenous or external. In this type of ictal transition, the seizure possibly may be anticipated in its early, preclinical phases. In the second case, where a sharp critical transition takes place, we can assume that the system has at least two simultaneous interictal and ictal attractors all the time. To which attractor the trajectories converge, depends on the initial conditions and the system's parameters. An essential question in this scenario is how the transition between the normal ongoing and the seizure activity takes place. Such a transition can occur either due to the influence of external or endogenous factors or due to a random perturbation and, thus, it will be unpredictable. These dynamical changes may not be detectable from the analysis of the ongoing EEG, but they may be observable only by measuring the system's response to externally administered stimuli. In the special cases of reflex epilepsy, the leap between the normal ongoing attractor and the ictal attractor is caused by a well-defined external perturbation. Examples from these different scenarios are presented and discussed.     

Metal partitioning in a sulfidic canal sediment: metal solubility as a function of pH combined with EDTA extraction in anoxic conditions

The chemical forms of low concentrations of metals (Zn, Pb, Cu, Cd, Co and Mn) and main components (Fe and Ca) were determined under the original reducing conditions of a sulfide-rich sediment from the Gent-Terneuzen Canal (Belgium). Therefore, dissolution experiments as a function of pH were made in a salt solution mimicking the canal water. The centrifugates remaining after the dissolution vs. pH experiments were subsequently further extracted with 0.1 M EDTA/0.5 M NaAc (pH 4.65) to determine eventually readsorbed and/or reprecipitated metal ions. The experimental dissolution vs. pH edge of calcium, iron, manganese and cobalt had a lower slope than theoretically expected on the basis of the solubility of, respectively, calcium carbonate, iron sulfide/iron carbonate, manganese sulfide/manganese carbonate and cobalt sulfide and was explained by the combination of (a) the solubilities of the various minerals and (b) metal readsorption onto clay minerals and organic matter. Higher metal recoveries were measured in the 0.1 M EDTA/0.5 M NaAc mixture and proved that in addition coprecipitate formation with iron sulfide/iron carbonate minerals may occur. The solubility of zinc, lead, cadmium and copper was very low in the mimicking salt solution even at very low pH values (up to pH 1) in agreement with the theoretical solubility of their discrete metal sulfides. However, by using an additional 0.1 M EDTA/0.5 M NaAc extraction on the centrifugates remaining after the dissolution vs. pH experiments, it was qualitatively shown that zinc and lead were partly associated with iron sulfide/iron carbonate phases in the real sediment (in addition to their presence in discrete metal sulfides). Cadmium was present solely in discrete cadmium sulfide phases. It was not possible to verify whether copper was present in discrete copper sulfide phases and/or in mixed coprecipitates with iron sulfide/iron carbonate minerals.     

Ultra-Low-Power Interface Chip for Autonomous Capacitive Sensor Systems

Traditionally, most of the sensor interfaces must be tailored towards a specific application. This approach results in a high recurrent design cost and time to market. On the other hand, generic sensor interface design reduces the costs and offers a handy solution for multisensor applications. This paper presents a generic sensor interface chip (GSIC), which can read out a broad range of capacitive sensors. It contains capacitance-to-voltage converters, a switched-capacitor amplifier, an analog-to-digital converter, oscillators, clock generation circuits and a reference circuit. The system combines a very low-power design with a smart energy management, which adapts the current consumption according to the accuracy and speed requirements of the application. The GSIC is used in a pressure and an acceleration monitoring system. The pressure monitoring system achieves a current drain of 2.3 muA for a 10-Hz sample frequency and an 8-bit accuracy. In the acceleration monitoring system, we measured a current of 3.3 muA for a sample frequency of 10 Hz and an accuracy of 9 bits     

CrimeSPOT: A language and runtime for developing active wireless sensor network applications

Advances in wireless sensing and actuation technology allow embedding significant amounts of application logic inside wireless sensor networks. Such active WSN applications are more autonomous, but are significantly more complex to implement. Event-based middleware lends itself to implementing these applications. It offers developers fine-grained control over how an individual node interacts with the other nodes of the network. However, this control comes at the cost of event handlers which lack composability and violate software engineering principles such as separation of concerns. In this paper, we present CrimeSPOT as a domain-specific language for programming WSN applications on top of event-driven middleware. Its node-centric features enable programming a node’s interactions through declarative rules rather than event handlers. Its network-centric features support reusing code within and among WSN applications. Unique to CrimeSPOT is its support for associating application-specific semantics with events that carry sensor readings. These preclude transposing existing approaches that address the shortcomings of event-based middleware to the domain of wireless sensor networks. We provide a comprehensive overview of the language and the implementation of its accompanying runtime. The latter comprises several extensions to the Rete forward chaining algorithm. We evaluate the expressiveness of the language and the overhead of its runtime using small, but representative active WSN applications.     

Multi-cue hand detection and tracking for a head-mounted augmented reality system

With the recent developments in wearable augmented reality (AR), the role of natural human–computer interaction is becoming more important. Utilization of auxiliary hardware for interaction introduces extra complexity, weight and cost to wearable AR systems and natural means of interaction such as gestures are therefore more desirable. In this paper, we present a novel multi-cue hand detection and tracking method for head-mounted AR systems which combines depth, color, intensity and curvilinearity. The combination of different cues increases the detection rate, eliminates the background regions and therefore increases the tracking performance under challenging conditions. Detected hand positions and the trajectories are used to perform actions such as click, select, etc. Moreover, the 6 DOF poses of the hands are calculated by approximating the segmented regions with planes in order to render a planar menu (interface) around the hand and use the hand as a planar selection tool. The proposed system is tested on different scenarios (including markers for reference) and the results show that our system can detect and track the hands successfully in challenging conditions such as cluttered and dynamic environments and illumination variance. The proposed hand tracker outperforms other well-known hand trackers under these conditions.