No abstract

Phospholipase A2 (PLA2) enzymes catalyze the hydrolysis of the sn-2 ester linkage of glycerophospholipids to produce fatty acids and lysophospholipids. A significant number of mammalian phospholipases comprise a family of secreted PLA2 enzymes, found in specific tissues and cellular locations, exhibiting unique enzymatic properties and distinct biological functions. Development of new real-time spectrofluorimetric PLA2 assays should facilitate the kinetic characterization and mechanistic elucidation of the isozymes in vitro, with the potential applicability to detect and measure catalytic PLA2 activity in tissues and cellular locations. Here we report a new synthesis of double-labeled phosphatidylcholine analogues with chain-terminal reporter groups including coumarin fluorophores for fluorescence resonance energy transfer (FRET)-based kinetic studies of PLA2 enzymes. The use of coumarin derivatives as fluorescent labels provides reporter groups with substantially decreased size compared to the first generation of donor-acceptor pairs of fluorescent phospholipids. The key advantage of the design is to interfere less with the physicochemical properties of the acyl chains, thereby improving the substrate quality of the synthetic probes. In order to assess the impact of the fluorophore substituents on the catalytic hydrolysis and on the phospholipid packing in the lipid-water interface of the assay, we used the experimentally determined specific activity of bee-venom phospholipase A2 as a model for the secretory PLA2 enzymes. Specifically, the rate of PLA2 hydrolysis of the coumarin labeled phosphatidylcholine analogues was less than three times slower than natural substrate dipalmitoyl phosphatidylcholine (DPPC) under the same experimental conditions. Furthermore, variation of the mole fraction of the synthetic phosphatidylcholine vs. that of the natural DPPC substrate showed nearly ideal mixing behavior in the phospholipid-surfactant aggregates of the assay. The synthesis provides a rapid and efficient method for preparation of new synthetic phosphatidylcholines with the desired target structures for enzymatic and physicochemical studies.

Price played an important role in most purchases. Buying behavior was strongly determined by consumers' price expectations. Emotion as a research hotspot was demonstrated to be ubiquitous in marketing and influenced purchase processing as well. This study addressed interests upon whether emotion arousal would influence consumers' price perceptions and their willingness to purchase. Compared to such emotion researches which normally adopted emotional pictures as priming stimuli, we creatively employed a two-player “Finger Play” (FP) game without monetary gains or losses to arouse subjects' emotion in the experiment. A 2 (FP Game Results: Continuous Win vs. Continuous Lose) by 2 (Price Conditions: High Price vs. Low Price) Event-Related Potentials (ERPs) experiment was designed to investigate whether game results would arouse different emotions and influence subjects' perception of product price. Both behavioral and ERP results indicated that subjects' price perception was deeply impacted by emotions induced from continuous win/lose experiences.

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Real-time flood control operations of a cascade reservoir system under emergency conditions can reduce the social and economic loss caused by natural disasters. This paper proposes a risk-based model for real-time flood control operation of reservoirs under emergency conditions and uncertainties. The proposed model consists of three modules: emergency scenarios establishing, Monte Carlo simulations, and risk analysis. The emergency scenarios considered are earthquakes, extreme floods and failure of the spillways of a reservoir. The uncertainty factor considered is the forecast error of reservoir inflows, arising from model structural uncertainty and parameter estimating. The Monte Carlo simulations conduct the real-time flood control operation of reservoirs considering the proposed emergency events and uncertainties. The module of risk analysis performs the assessment of the operation schedules and calculates the risk of dam overtopping, based on the results from Monte Carlo simulations. The proposed model is applied to a cascade reservoir system in the upper reaches of Daduhe river basin in China. The results show that the maximum initial water level of the Shuangjiangkou reservoir is 2447 m a.s.l. (meters above sea level) using the release capacity model and is 2444.5 m a.s.l. using the command model under the scenario of upstream dam break. The integrated risk of the reservoir increases with the initial water level and the uncertainty degree of the reservoir inflows. The decision-makers can choose the operation models according to the actual initial water level of the reservoir under different emergency scenarios.