RhoA/ROCK-dependent moesin phosphorylation regulates AGE-induced endothelial cellular response

<p>Abstract</p> <p>Background</p> <p>The role of advanced glycation end products (AGEs) in the development of diabetes, especially diabetic complications, has been emphasized in many reports. Accumulation of AGEs in the vasculature triggers a series of morphological and functional changes in endothelial cells (ECs) and induces an increase of endothelial permeability. This study was to investigate the involvement of RhoA/ROCK-dependent moesin phosphorylation in endothelial abnormalities induced by AGEs.</p> <p>Methods</p> <p>Using human dermal microvascular endothelial cells (HMVECs), the effects of human serum albumin modified-AGEs (AGE-HSA) on the endothelium were assessed by measuring monolayer permeability and staining of F-actin in HMVECs. Activations of RhoA and ROCK were determined by a luminescence-based assay and immunoblotting. Transfection of recombinant adenovirus that was dominant negative for RhoA (RhoA N19) was done to down-regulate RhoA expression, while adenovirus with constitutively activated RhoA (RhoA L63) was transfected to cause overexpression of RhoA in HMVECs. H-1152 was employed to specifically block activation of ROCK. Co-immunoprecipitation was used to further confirm the interaction of ROCK and its downstream target moesin. To identify AGE/ROCK-induced phosphorylation site in moesin, two mutants pcDNA3/HA-moesinT^558Aand pcDNA3/HA-moesinT^558Dwere applied in endothelial cells.</p> <p>Results</p> <p>The results showed that AGE-HSA increased the permeability of HMVEC monolayer and triggered the formation of F-actin-positive stress fibers. AGE-HSA enhanced RhoA activity as well as phosphorylation of ROCK in a time- and dose-dependent manner. Down-regulation of RhoA expression with RhoA N19 transfection abolished these AGE-induced changes, while transfection of RhoA L63 reproduced the AGE-evoked changes. H-1152 attenuated the AGE-induced alteration in monolayer permeability and cytoskeleton. The results also confirmed the AGE-induced direct interaction of ROCK and moesin. Thr558 was further identified as the phosphorylating site of moesin in AGE-evoked endothelial responses.</p> <p>Conclusion</p> <p>These results confirm the involvement of RhoA/ROCK pathway and subsequent moesin Thr558 phosphorylation in AGE-mediated endothelial dysfunction.</p

Phylogenetic, Expression, and Bioinformatic Analysis of the ABC1

We studied 17 ABC1 genes in Populus trichocarpa, all of which contained an ABC1 domain consisting of about 120 amino acid residues. Most of the ABC1 gene products were located in the mitochondria or chloroplasts. All had a conserved VAVK-like motif and a DFG motif. Phylogenetic analysis grouped the genes into three subgroups. In addition, the chromosomal locations of the genes on the 19 Populus chromosomes were determined. Gene structure was studied through exon/intron organization and the MEME motif finder, while heatmap was used to study the expression diversity using EST libraries. According to the heatmap, PtrABC1P14 was highlighted because of the high expression in tension wood which related to secondary cell wall formation and cellulose synthesis, thus making a contribution to follow-up experiment in wood formation. Promoter cis-element analysis indicated that almost all of the ABC1 genes contained one or two cis-elements related to ABA signal transduction pathway and drought stress. Quantitative real-time PCR was carried out to evaluate the expression of all of the genes under abiotic stress conditions (ABA, CdCl2, high temperature, high salinity, and drought); the results showed that some of the genes were affected by these stresses and confirmed the results of promoter cis-element analysis

Pressure induced superconductivity bordering a charge-density-wave state in NbTe4 with strong spinorbit coupling

Transition-metal chalcogenides host various phases of matter, such as charge-density wave (CDW), superconductors, and topological insulators or semimetals. Superconductivity and its competition with CDW in low-dimensional compounds have attracted much interest and stimulated considerable research. Here we report pressure induced superconductivity in a strong spin-orbit (SO) coupled quasi-one-dimensional (1D) transition-metal chalcogenide NbTe$_4$, which is a CDW material under ambient pressure. With increasing pressure, the CDW transition temperature is gradually suppressed, and superconducting transition, which is fingerprinted by a steep resistivity drop, emerges at pressures above 12.4 GPa. Under pressure $p$ = 69 GPa, zero resistance is detected with a transition temperature $T_c$ = 2.2 K and an upper critical field $H_{c2}$= 2 T. We also find large magnetoresistance (MR) up to 102\% at low temperatures, which is a distinct feature differentiating NbTe$_4$ from other conventional CDW materials.Comment: https://rdcu.be/LX8

Quantum Well Laser Diodes With Slightly-Doped Tunnel Junction

We experimentally investigate the electrical and optical characteristics of conventional quantum well laser diodes and the quantum well laser diodes with slightly-doped tunnel junction N++GaAs/undoped-GaAs. The results show that the slightly-doped tunnel junction give significant role on the laser diodes performances in the InGaAs/GaAs quantum well material system. The TJ LD has a internal quantum efficiency of 21% and the loss is 6.9 em -1 , the current threshold is 35 mA, both the lasers are operating at 1.06 μm, but the slightly-doped tunnel junction diode show nonlinear S-shaped current-voltage and broadband lasing characteristics. The results may also lead to the realization of more applications

Asymmetric quantum well broadband thyristor laser

A broadband thyristor laser based on InGaAs/GaAs asymmetric quantum well (AQW) is fabricated by metal organic chemical vapor deposition (MOCVD). The 3-μm-wide Fabry–Perot (FP) ridge-waveguide laser shows an S-shape I−V characteristic and exhibits a flat-topped broadband optical spectrum coverage of ~27 nm (Δ−10 dB) at a center wavelength of ~1090 nm with a total output power of 137 mW under pulsed operation. The AQW structure was carefully designed to establish multiple energy states within, in order to broaden the gain spectrum. An obvious blue shift emission, which is not generally acquired in QW laser diodes, is observed in the broadening process of the optical spectrum as the injection current increases. This blue shift spectrum broadening is considered to result from the prominent band-filling effect enhanced by the multiple energy states of the AQW structure, as well as the optical feedback effect contributed by the thyristor laser structure

Development of a Computerized Adaptive Testing for Internet Addiction

Internet addiction disorder has become one of the most popular forms of addiction in psychological and behavioral areas, and measuring it is growing increasingly important in practice. This study aimed to develop a computerized adaptive testing to measure and assess internet addiction (CAT-IA) efficiently. Four standardized scales were used to build the original item bank. A total of 59 polytomously scored items were finally chosen after excluding 42 items for failing the psychometric evaluation. For the final 59-item bank of CAT-IA, two simulation studies were conducted to investigate the psychometric properties, efficiency, reliability, concurrent validity, and predictive validity of CAT-IA under different stopping rules. The results showed that (1) the final 59 items met IRT assumptions, had high discrimination, showed good item-model fit, and were without DIF; and (2) the CAT-IA not only had high measurement accuracy in psychometric properties but also sufficient efficiency, reliability, concurrent validity, and predictive validity. The impact and limitations of CAT-IA were discussed, and several suggestions for future research were provided

Ultrasound microbubble-mediated delivery of the siRNAs targeting MDR1 reduces drug resistance of yolk sac carcinoma L2 cells

<p>Abstract</p> <p>Background</p> <p>MDR1 gene encoding P-glycoprotein is an ATP-dependent drug efflux transporter and related to drug resistance of yolk sac carcinoma. Ultrasound microbubble-mediated delivery has been used as a novel and effective gene delivery method. We hypothesize that small interfering RNA (siRNA) targeting MDR1 gene (siMDR1) delivery with microbubble and ultrasound can down-regulate MDR1 expression and improve responsiveness to chemotherapeutic drugs for yolk sac carcinoma <it>in vitro</it>.</p> <p>Methods</p> <p>Retroviral knockdown vector pSEB-siMDR1s containing specific siRNA sites targeting rat MDR1 coding region were constructed and sequence verified. The resultant pSEB-siMDR1 plasmids DNA were encapsulated with lipid microbubble and the DNA release were triggered by ultrasound when added to culture cells. GFP positive cells were counted by flow cytometry to determine transfection efficiency. Quantitative real-time PCR and western blot were performed to determine the mRNA and protein expression of MDR1. P-glycoprotein function and drug sensitivity were analyzed by Daunorubicin accumulation and MTT assays.</p> <p>Results</p> <p>Transfection efficiency of pSEB-siMDR1 DNA was significantly increased by ultrasound microbubble-mediated delivery in rat yolk sac carcinoma L2 (L2-RYC) cells. Ultrasound microbubble-mediated siMDR1s delivery effectively inhibited MDR1 expression at both mRNA and protein levels and decreased P-glycoprotein function. Silencing MDR1 led to decreased cell viability and IC₅₀of Vincristine and Dactinomycin.</p> <p>Conclusions</p> <p>Our results demonstrated that ultrasound microbubble-mediated delivery of MDR1 siRNA was safe and effective in L2-RYC cells. MDR1 silencing led to decreased P-glycoprotein activity and drug resistance of L2-RYC cells, which may be explored as a novel approach of combined gene and chemotherapy for yolk sac carcinoma.</p