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.     

A new method for modeling preoperative diagnosis of ovarian tumors

In this paper, we present a sequential nonuniform procedure, an inference method which combines feature selection based on the Kullback information gain and a step-wise classification procedure to produce a reliable, interpretable, and robust model. We applied the model to an ovarian tumor data set to distinguish between malignant and benign tumors. The performance of the model was assessed using receiver operating characteristic (ROC) analysis and gave an overall accuracy over 85%, and area under the curve (AUC) of 0.887 which compares well with existing methods. The method presented here is significant because of its ability to handle missing values, and it only uses a small number of variables which are graded according to their discriminative relevance. This, together with the fact that the resulting model is interpretable and has good performance, is likely to lead to widespread clinical acceptance of the method. The method is also generic and can be readily adapted for other classifications problems in biomedicine.     

Market power mitigation by regulating contract portfolio risk

Abuse of market power by dominant generation firms is a growing concern in worldwide electricity markets. This paper argues that relying only on general competition rules—as is the case in most European countries—is insufficient and that complementary ex-ante regulation is needed. In particular, regulators should incentivize firms to sign contracts with retailers by regulating their risk exposure. In a simulation model we show that this type of regulation can significantly reduce the deadweight loss in the market, without imposing large costs on regulatees.     

Exploring time series retrieved from cardiac implantable devices for optimizing patient follow-up

Current cardiac implantable devices (IDs) are equipped with a set of sensors that can provide useful information to improve patient follow-up and prevent health deterioration in the postoperative period. In this paper, data obtained from an ID with two such sensors (a transthoracic impedance sensor and an accelerometer) are analyzed in order to evaluate their potential application for the follow-up of patients treated with a cardiac resynchronization therapy (CRT). A methodology combining spatiotemporal fuzzy coding and multiple correspondence analysis (MCA) is applied in order to: 1) reduce the dimensionality of the data and provide new synthetic indexes based on the "factorial axes" obtained from MCA; 2) interpret these factorial axes in physiological terms; and 3) analyze the evolution of the patient's status by projecting the acquired data into the plane formed by the first two factorial axes named "factorial plane." In order to classify the different evolution patterns, a new similarity measure is proposed and validated on the simulated datasets, and then, used to cluster observed data from 41 CRT patients. The obtained clusters are compared with the annotations on each patient's medical record. Two areas on the factorial plane are identified, one being correlated with a health degradation of patients and the other with a stable clinical state.     

Effect of propionic acid on kinetics of acetate and oleate and on plasma and milk fatty acid composition of goats

Entry rates of acetate and oleate and their incorporation into lipids of blood plasma and fatty acids of milk were studied in lactating goats fed a concentrate-roughage ration with propionic acid infused intraruminally at 0, 5.52, and 13.74 g/h by primed constant intravenous infusion of [1-carbon-14] acetate and [9, 10-hydrogen-3] oleate. Means for infusion rates were acetate, 60, 52, and 39 micrograms/ml blood plasma; propionate 9, 12, and 22 micrograms/ml; oleate, 19, 14, and 12 micrograms/ml; acetate entry rate, 3.9, 2.7, and 1.8 mmol/h per kg bodyweight; oleate entry rate, 47, 29, and 19 mumol/h per kg bodyweight; acetate oxidation rate, 2.0, 1.7, and 1.4 mmol/h per kg, and its contribution to the total carbon dioxide production, 16, 14, and 11%. Propionic acid increased incorporation of carbon-14 and hydrogen-3 into plasma lipids, elevated proportions of 7:0, 9:0, 11:0, 13:0, 15:0, and 17:0 fatty acids in milk, and tended to lower others. Specific radioactivities of milk fatty acids during infusion of propionic acid were elevated by 1.8 to 2.8 times, and total fatty acids in milk and plasma were lowered by 22 and 38%. Data support the glucogenic theory that propionic acid either directly or through gluconeogenesis stimulates insulin secretion, which in turn inhibits release of fatty acids from adipose tissue, resulting in milk fat depression.     

Experimental investigation on the combustion behaviour of pre-dried Greek lignite

The present paper includes the results of the combustion tests with Greek dried lignite performed at a 1 MWth semi industrial scale pulverized coal combustion facility. Scope of the campaign is the investigation of the combustion behaviour of Greek lignite, i.e. temperature fields, ignition, burnout, emissions, as well as slagging and fouling tendency, while firing with varying levels of recirculated flue gas. Dry coal co-firing conditions in a large scale boiler are simulated by adjusting the volume flow of recirculated flue gas.Two test series representing different boiler operation modes are performed. During the first series the maximum flue gas temperature increase, when co-firing dry coal, is determined, while in the second test series the needed load decrease, in order to keep constant furnace outlet temperature in dry coal co-firing conditions is recorded. A detailed measurement set is carried out including temperature profiles, emissions, fuel, fly ash sampling and slagging and fouling investigations through the installation of dedicated deposition probes.The anticipated increase of the furnace temperature profiles by decreasing the inserted recirculated flue gas is confirmed by the experimental results. No clear trend of dry coal co-combustion on the emissions' behaviour is noticed, while dry coal firing appears to have a moderate effect on the deposition behaviour of Greek lignite. These preliminary investigations indicate that no significant operational problems are expected during a potential future demonstration of dry lignite co-firing in a Greek large scale boiler.     

Reynolds stress and turbulence estimates in bottom boundary layer of Fall of Warness

A broadband acoustic Doppler current profiler (ADCP) moored on the seabed at 42 m depth has been used to observe the mean and turbulent flow components in the tidally energetic Fall of Warness channel over two tidal cycles. The Reynolds stress has been estimated from the difference in variance between the along-beam velocities of opposing acoustic beams. Near bed stress at 2.63 m above seabed (mab) exceeds 7.5 Pa at the time of mean flow (speed of ~ 1.3 m s^− 1) while the ebb stresses are limited to ~ 3.31 Pa during the peak ebb, mean, flow of ~ 1.3 m s^− 1. The production of turbulent kinetic energy (TKE), P was found to be negative below 2 × 10^− 9 W m^− 3 and up to 6 × 10^− 4 W m^− 3 was estimated during flood flows and decreasing to 3 × 10^− 4 W m^− 3. The TKE dissipation rate ε was estimated by inertial dissipation method (IDM) with the greatest value of 2.43 × 10^− 2 W m^− 3 observed near the seabed around maximum ebb, falling to 5.75 × 10^− 5 W m^− 3 around slack water. The comparison between P and ε was performed by calculating individual ratios of P corresponding to ε using a bootstrap resampling technique. The study shows that the ratio ε/P averaged over whole flood and ebb were found to be ~ 0.4138 and ~ 0.4177, respectively, indicating that production exceeded dissipation. The uncertainties in Reynolds stress estimates due to instrument noise were found to be 3 × 10^− 4 Pa while 4.52 × 10^− 2 Pa can be attributed to the uncertainties due to the increase in the flow-related component.