Identification of Sortilin Alternatively Spliced Variants in Mouse 3T3L1 Adipocytes

Type 2 diabetes mellitus is a metabolic disorder defined by systemic insulin resistance. Insulin resistance in adipocytes, an important regulator of glucose metabolism, results in impaired glucose uptake. The trafficking protein, sortilin, regulates major glucose transporter 4 (Glut4) movement, thereby promoting glucose uptake in adipocytes. Here, we demonstrate the presence of an alternatively spliced sortilin variant (Sort^17b), whose levels increase with insulin resistance in mouse 3T3L1 adipocytes. Using a splicing minigene, we show that inclusion of alternative exon 17b results in the expression of Sort^17b splice variant. Bioinformatic analysis indicated a novel intrinsic disorder region (IDR) encoded by exon 17b of Sort^17b. Root mean square deviation (RMSD) and root mean square fluctuation (RMSF) measurements using molecular dynamics demonstrated increased flexibility of the protein backbone within the IDR. Using protein–protein docking and co-immunoprecipitation assays, we show robust binding of Glut4 to Sort^17b. Further, results demonstrate that over-expression of Sort^17b correlates with reduced Glut4 translocation and decreased glucose uptake in adipocytes. The study demonstrates that insulin resistance in 3T3L1 adipocytes promotes expression of a novel sortilin splice variant with thus far unknown implications in glucose metabolism. This knowledge may be used to develop therapeutics targeting sortilin variants in the management of type 2 diabetes and metabolic syndrome.     

Microtubule Destabilizing Sulfonamides as an Alternative to Taxane-Based Chemotherapy

Pan-Gyn cancers entail 1 in 5 cancer cases worldwide, breast cancer being the most commonly diagnosed and responsible for most cancer deaths in women. The high incidence and mortality of these malignancies, together with the handicaps of taxanes—first-line treatments—turn the development of alternative therapeutics into an urgency. Taxanes exhibit low water solubility that require formulations that involve side effects. These drugs are often associated with dose-limiting toxicities and with the appearance of multi-drug resistance (MDR). Here, we propose targeting tubulin with compounds directed to the colchicine site, as their smaller size offer pharmacokinetic advantages and make them less prone to MDR efflux. We have prepared 52 new Microtubule Destabilizing Sulfonamides (MDS) that mostly avoid MDR-mediated resistance and with improved aqueous solubility. The most potent compounds, N-methyl-N-(3,4,5-trimethoxyphenyl-4-methylaminobenzenesulfonamide 38, N-methyl-N-(3,4,5-trimethoxyphenyl-4-methoxy-3-aminobenzenesulfonamide 42, and N-benzyl-N-(3,4,5-trimethoxyphenyl-4-methoxy-3-aminobenzenesulfonamide 45 show nanomolar antiproliferative potencies against ovarian, breast, and cervix carcinoma cells, similar or even better than paclitaxel. Compounds behave as tubulin-binding agents, causing an evident disruption of the microtubule network, in vitro Tubulin Polymerization Inhibition (TPI), and mitotic catastrophe followed by apoptosis. Our results suggest that these novel MDS may be promising alternatives to taxane-based chemotherapy in chemoresistant Pan-Gyn cancers.     

Inhibition of RANKL-Induced Osteoclastogenesis by Novel Mutant RANKL

Background: Recently, it was reported that leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4, also called GPR48) is another receptor for RANKL and was shown to compete with RANK to bind RANKL and suppress canonical RANK signaling during osteoclast differentiation. The critical role of the protein triad RANK–RANKL in osteoclastogenesis has made their binding an important target for the development of drugs against osteoporosis. In this study, point-mutations were introduced in the RANKL protein based on the crystal structure of the RANKL complex and its counterpart receptor RANK, and we investigated whether LGR4 signaling in the absence of the RANK signal could lead to the inhibition of osteoclastogenesis.; Methods: The effects of point-mutated RANKL (mRANKL-MT) on osteoclastogenesis were assessed by tartrate-resistant acid phosphatase (TRAP), resorption pit formation, quantitative real-time polymerase chain reaction (qPCR), western blot, NFATc1 nuclear translocation, micro-CT and histomorphological assay in wild type RANKL (mRANKL-WT)-induced in vitro and in vivo experimental mice model. Results: As a proof of concept, treatment with the mutant RANKL led to the stimulation of GSK-3β phosphorylation, as well as the inhibition of NFATc1 translocation, mRNA expression of TRAP and OSCAR, TRAP activity, and bone resorption, in RANKL-induced mouse models; and Conclusions: The results of our study demonstrate that the mutant RANKL can be used as a therapeutic agent for osteoporosis by inhibiting RANKL-induced osteoclastogenesis via comparative inhibition of RANKL. Moreover, the mutant RANKL was found to lack the toxic side effects of most osteoporosis treatments.     

基于可调谐半导体激光吸收光谱技术的甲烷气体监测研究

设计了一种基于可调谐半导体激光吸收光谱技术的甲烷(CH4)气体监测装置,该装置具有温度监测范围大、监测精度高和实时监测的优点。监测装置的激光光源为波长1650 nm的反馈式激光器,应用波长调谐与锁相放大器技术,对周围环境进行温度补偿与背景扣除,可以较准确地测量周围环境中CH4的体积分数。实验表明,当CH4体积分数小于1%时,该装置的监测精度为±0.02%;当CH4体积分数大于1%时,装置监测误差小于±0.8%,实时监测的最长响应时间为10 s。装置的温度范围为0~40℃,可满足大多数工业生产中对CH4等气体体积分数的监测需求。     

Eddy Current Detection of Retained Austenite in Ni-Hard4 Cast Iron

In this article, the nondestructive eddy current (EC) method was used for microstructural evaluation of Ni-hard4 cast iron (NiHCI). Different destabilizing heat treatments were used to produce different microstructures and hardness. The microstructural analysis and hardness measurement were done for conventional characterization of the reference blocks. Then, the EC techniques were applied to evaluate the hardness and microstructural changes by the detection of EC parameters at different frequencies. Metallurgical investigations showed that increasing the destabilizing soaking time and temperature decreases the amount of retained austenite in NiHCIs. The nondestructive evaluation method (NDT) revealed that it can be good correlated between retained austenite amount and hardness with EC parameters. This shows that the EC technique can detect NiHCIs products according to the metallurgical characterization with good accuracy.     

FDM用PETG系列耗材的改性研究

对市售聚对苯二甲酸乙二醇酯-1，4-环己烷二甲醇酯（PETG）丝材及原材料进行研究，选用PETG 2012作为制备熔融堆积成型技术（FDM）用PETG丝材的基体树脂。选用聚碳酸酯（PC）对基体PETG进行增韧增强改性，并制成3D打印高分子丝材进行打印测试，探讨了PC含量对PETG耗材及制件性能的影响。结果表明，在低PC含量下，PETG与PC的相容性较好，PETG与PC制件未出现明显的分层，分布比较均匀；共混物的流动性随着PC含量的增加而降低，且打印件的维卡软化温度和热变形温度随着PC用量的增加而增加；此外，PC材料的加入使得PETG打印件的力学性能得到较大改善：打印件的拉伸强度随着PC用量的增加而增大，缺口冲击强度却随着PC用量的增加呈现先增加后减小的趋势； PETG/PC的配比为1∶1时，可制得力学性能优异的3D打印耗材。     

Hydrogen embrittlement of the coarse grain heat affected zone of a quenched and tempered 42CrMo4 steel

The coarse grain heat affected zone (CG-HAZ) of welds produced in a quenched and tempered 42CrMo4 steel was simulated by means of a laboratory heat treatment consisting in austenitizing at 1200 °C for 20 min, oil quenching and finally applying a post weld heat treatment at 700 °C for 2 h (similar to the tempering treatment previously applied to the base steel). A tempered martensite microstructure with a prior austenite grain size of 150 μm and a hardness of 230 HV, similar to the aforementioned CG-HAZ weld region, was produced. The effect of the prior austenite grain size on the hydrogen embrittlement (HE) behaviour of the steel was studied comparing this coarse-grained microstructure with that of the fine-grained base steel, with a prior austenite grain size of 20 μm.The specimens used in this study were charged with hydrogen gas in a reactor at 19.5 MPa and 450 °C for 21 h. Cylindrical specimens were used to determine hydrogen uptake and hydrogen desorption behaviour. Smooth and notched tensile specimens tested under different displacement rates were also used to evaluate HE.Embrittlement indexes, EI, were generally quite low in the case of hydrogen pre-charged tensile tests performed on smooth tensile specimens. However, very significant embrittlement indexes were obtained with notched tensile specimens. It was observed that these indexes always increase as the applied displacement rate decreases. Moreover, hydrogen embrittlement indexes also increase with increasing prior austenite grain size. In fact, the embrittlement index related to the reduction in area, EI_(RA), reached values of over 20% and 50% for the fine and coarse grain size steels, respectively, when tested under the lowest displacement rates (0.002 mm/min).A comprehensive fractographic analysis was performed and the main operative failure micromechanisms due to the presence of internal hydrogen were determined at different test displacement rates. While microvoids coalescence (MVC) was found to be the typical ductile failure micromechanism in the absence of hydrogen in the two steels, brittle decohesion mechanisms (carbide-matrix interface decohesion, CMD, and martensitic lath interface decohesion, MLD) were observed under internal hydrogen. Intergranular fracture (IG) was also found to be operative in the case of the coarse-grained steel tested under the lowest displacement rate, in which hydrogen accumulation in the process zone ahead of the notch tip is maximal.     

Improved multi-recessed 4H–SiC MESFETs with double-recessed p-buffer layer

An improved multi-recessed 4H–SiC metal semiconductor field effect transistor (MRD-MESFET) with double-recessed p-buffer layer (DRB-MESFET) is proposed in this paper. By introducing a double-recessed p-buffer layer, the gate depletion layer is further modulated, and higher drain saturation current and DC transconductance are obtained compared with the MRD-MESFET. The simulations show that the drain saturation current of the DRB-MESFET is about 42.4% larger than that of the MRD-MESFET. The DC transconductance of the DRB-MESFET is almost 15% higher than that of the MRD-MESFET and very close to that of double-recessed structure (DR-MESFET) at the bias conditions of V_gs=0 V and V_ds=40 V. The proposed structure has an improvement of 26.1% and 74.2% in the output maximum power density compared with that of the MRD-MESFET and DR-MESFET, respectively. In the meanwhile, the proposed structure possesses smaller gate-source capacitance, which results in better RF characteristics.     

Foodborne transmission of Listeria monocytogenes via ready-to-eat salad: A nationwide outbreak in Switzerland, 2013–2014

From 26 October 2013 to 23 April 2014, 32 cases of listeriosis infected with an Listeria monocytogenes strain serovar 4b, sequence type 4 and belonging to a single distinct PFGE pulsotype were registered in patients from several cantons of Switzerland. L. monocytogenes was detected in blood (75%), CSF (16%), ascites (6%) and in joint fluid (3%) samples. By the end of March 2014, a food producing company reported an L. monocytogenes contamination of ready-to-eat salads to the authorities after detecting the pathogen through its in-house routine quality control. Product and environmental samples collected during subsequent investigations yielded isolates, matching the outbreak strain, thus confirming that ready-to-eat salad from this company was most likely the outbreak source. The cause for the product contamination was related to a design-inherent hygienic problem of one specific product-feeding belt. Complementary patient interviews also identified ready-to-eat green salads bought at one retailer as the likely outbreak source.