Endothelial Cell-Specific Molecule 2 (Ecsm2) Localizes To Cell-Cell Junctions And Modulates Bfgf-Directed Cell Migration Via The Erk-Fak Pathway

Background: Despite its first discovery by in silico cloning of novel endothelial cell-specific genes a decade ago, the biological functions of endothelial cell-specific molecule 2 (ECSM2) have only recently begun to be understood. Limited data suggest its involvement in cell migration and apoptosis. However, the underlying signaling mechanisms and novel functions of ECSM2 remain to be explored. Methodology/Principal Findings: A rabbit anti-ECSM2 monoclonal antibody (RabMAb) was generated and used to characterize the endogenous ECSM2 protein. Immunoblotting, immunoprecipitation, deglycosylation, immunostaining and confocal microscopy validated that endogenous ECSM2 is a plasma membrane glycoprotein preferentially expressed in vascular endothelial cells (ECs). Expression patterns of heterologously expressed and endogenous ECSM2 identified that ECSM2 was particularly concentrated at cell-cell contacts. Cell aggregation and transwell assays showed that ECSM2 promoted cell-cell adhesion and attenuated basic fibroblast growth factor (bFGF)-driven EC migration. Gain or loss of function assays by overexpression or knockdown of ECSM2 in ECs demonstrated that ECSM2 modulated bFGF-directed EC motility via the FGF receptor (FGFR)-extracellular regulated kinase (ERK)-focal adhesion kinase (FAK) pathway. The counterbalance between FAK tyrosine phosphorylation (activation) and ERK-dependent serine phosphorylation of FAK was critically involved. A model of how ECSM2 signals to impact bFGF/FGFR-driven EC migration was proposed. Conclusions/Significance: ECSM2 is likely a novel EC junctional protein. It can promote cell-cell adhesion and inhibit bFGF-mediated cell migration. Mechanistically, ECSM2 attenuates EC motility through the FGFR-ERK-FAK pathway. The findings suggest that ECSM2 could be a key player in coordinating receptor tyrosine kinase (RTK)-, integrin-, and EC junctional component-mediated signaling and may have important implications in disorders related to endothelial dysfunction and impaired EC junction signaling. © 2011 Shi et al

Immunogenicity and Protective Efficacy against Murine Tuberculosis of a Prime-Boost Regimen with BCG and a DNA Vaccine Expressing ESAT-6 and Ag85A Fusion Protein

Heterologous prime-boost regimens utilizing BCG as a prime vaccine probably represent the best hope for the development of novel tuberculosis (TB) vaccines. In this study, we examined the immunogenicity and protective efficacy of DNA vaccine (pcD685A) expressing the fusion protein of Ag85A and ESAT-6 (r685A) and its booster effects in BCG-immunized mice. The recombinant r685A fusion protein stimulated higher level of antigen-specific IFN-γ release in tuberculin skin test- (TST-) positive healthy household contacts of active pulmonary TB patients than that in TST-negative population. Vaccination of C57BL/6 mice with pcD685A resulted in significant protection against challenge with virulent Mycobacterium tuberculosis H37Rv when compared with the control group. Most importantly, pcD685A could act as a BCG booster and amplify Th1-type cell-mediated immunity in the lung of BCG-vaccinated mice as shown the increased expression of IFN-γ. The most significant reduction in bacterial load of both spleen and lung was obtained in mice vaccinated with BCG prime and pcD685A DNA booster when compared with BCG or pcD685A alone. Thus, our study indicates that pcD685A may be an efficient booster vaccine against TB with a strong ability to enhance prior BCG immunity

Protective Role of Nuclear Factor E2-Related Factor 2 against Acute Oxidative Stress-Induced Pancreatic β

Oxidative stress is implicated in the pathogenesis of pancreatic β-cell dysfunction that occurs in both type 1 and type 2 diabetes. Nuclear factor E2-related factor 2 (NRF2) is a master regulator in the cellular adaptive response to oxidative stress. The present study found that MIN6 β-cells with stable knockdown of Nrf2 (Nrf2-KD) and islets isolated from Nrf2-knockout mice expressed substantially reduced levels of antioxidant enzymes in response to a variety of stressors. In scramble MIN6 cells or wild-type islets, acute exposure to oxidative stressors, including hydrogen peroxide (H2O2) and S-nitroso-N-acetylpenicillamine, resulted in cell damage as determined by decrease in cell viability, reduced ATP content, morphology changes of islets, and/or alterations of apoptotic biomarkers in a concentration- and/or time-dependent manner. In contrast, silencing of Nrf2 sensitized MIN6 cells or islets to the damage. In addition, pretreatment of MIN6 β-cells with NRF2 activators, including CDDO-Im, dimethyl fumarate (DMF), and tert-butylhydroquinone (tBHQ), protected the cells from high levels of H2O2-induced cell damage. Given that reactive oxygen species (ROS) are involved in regulating glucose-stimulated insulin secretion (GSIS) and persistent activation of NRF2 blunts glucose-triggered ROS signaling and GSIS, the present study highlights the distinct roles that NRF2 may play in pancreatic β-cell dysfunction that occurs in different stages of diabetes

Forecasting Collector Road Speeds Under High Percentage of Missing Data

Accurate road speed predictions can help drivers in smart route planning. Although the issue has been studied previously, most existing work focus on arterial roads only, where sensors are configured closely for collecting complete real-time data. For collector roads where sensors sparsly cover, however, speed predictions are often ignored. With GPS-equipped floating car signals being available nowadays, we aim at forecasting collector road speeds by utilizing these signals. The main challenge compared with arterial roads comes from the missing data. In a time slot of the real case, over 90% of collector roads cannot be covered by enough floating cars. Thus most traditional approaches for arterial roads, relying on complete historical data, cannot be employed directly. Aiming at solving this problem, we propose a multi-view road speed prediction framework. In the first view, temporal patterns are modeled by a layered hidden Markov model; and in the second view, spatial patterns are modeled by a collective matrix factorization model. The two models are learned and inferred simultaneously in a co-regularized manner. Experiments conducted in the Beijing road network, based on 10K taxi signals in 2 years, have demonstrated that the approach outperforms traditional approaches by 10% in MAE and RMSE

Large scale blast emulator based explosive gas loading methods for structures and recent advances in experimental studies

This paper discusses the methods of utilizing acetylene-air mixture explosion for blast loading realization and its related experimental implementations, as well as the numerical simulation on comparison of dynamite explosion and acetylene-air mixture explosion based on LS-DYNA nonlinear explicit dynamic codes. The characteristic parameters of shock wave, such as over pressure and arriving time with respect to propagation distance, are compared for different blast cases. Then, the blast heat based equivalent principle and modified index of scale distance for gas explosion is proposed from the point view of energy conservation. The analysis results show that the Euler algorithm is useful for the numerical simulation of shock wave propagation caused by acetylene-air mixture explosion, and the results are in good accordance with the prediction results through different empirical formulations. With the increasing of propagation distance, the relative error between the two explosion methods is shown to be decreased. As a result, the explosive acetylene-air mixture can be used to conditionally realize the explosion effect of traditional chemical dynamites, such as TNT explosives. The results establish a theoretical foundation for using such explosive gas material in generating shock wave loadings based on the large scale blast emulator facility. At last, some preliminary test results on Tension Membrane structure, scaled steel frame structure and model energy resources structure are summarized based on this blast test method

Determination of oxytetracycline, tetracycline and chloramphenicol antibiotics in animal feeds using subcritical water extraction and high performance liquid chromatography

A rapid analytical method for the determination of oxytetracycline (OTC), tetracycline (TC) and chloramphenicol (CAP) antibiotics in animal feeds has been developed based on subcritical water extraction (SWE) without further sample clean-up followed by high performance liquid chromatography (HPLC) with ultraviolet (UV) detection. On extracting target antibiotics from spiked samples, the efficiency of the water extraction device was evaluated in terms of pH and volume of the extractant, temperature and time of the static extraction. The best extraction conditions were obtained by using 5.5mL of water adjusted to pH 2 with hydrochloric acid as the extractant at 100°C with 5-min static extraction. After filtration, 20μL of the aqueous extract was directly injected into the HPLC column. Recoveries between 82.1% and 90.0% with relative standard deviations ranging between 1.6% and 4.8% were achieved from spiked animal feed samples by using this method. Compared with the traditional ultrasonic extraction, this procedurewas remarkablymore efficient in extracting OTC, TC and CAP, simpler to perform, and there was no use of toxic organic solvents

Axial Thrust, Disk Frictional Losses, and Heat Transfer in a Gas Turbine Disk Cavity

The gas turbine is a kind of high-power and high-performance energy machine. Currently, it is a hot issue to improve the efficiency of the gas turbines by reducing the amount of secondary air used in the disk cavity. The precondition is to understand the effects of the through-flow rate on the axial thrust, the disk frictional losses, and the characteristics of heat transfer under various experimental conditions. In this paper, experiments are conducted to analyze the characteristics of flow and heat transfer. To ensure the safe operation of the gas turbine, the pressure distribution and the axial thrust are measured for various experimental conditions. The axial thrust coefficient is found to decrease as the rotational speed and the through-flow rate increases. By torque measurements, the amounts of the moment coefficient drop as the rotational speed increases while increase with through-flow rate. In order to better analyze the temperature field within the cavity, both the local and the average Nusselt number are investigated with the help of thermochromic liquid crystal technique. Four correlations for the local Nusselt number are determined according to the amounts of a through-flow coefficient. The results in this study can help the designers to better design the secondary air system in a gas turbine