Trypsiligase-Catalyzed Labeling of Proteins on Living Cells

Fluorescent fusion proteins are powerful tools for studying biological processes in living cells, but universal application is limited due to the voluminous size of those tags, which might have an impact on the folding, localization or even the biological function of the target protein. The designed biocatalyst trypsiligase enables site-directed linkage of small-sized fluorescence dyes on the N terminus of integral target proteins located in the outer membrane of living cells through a stable native peptide bond. The function of the approach was tested by using the examples of covalent derivatization of the transmembrane proteins CD147 as well as the EGF receptor, both presented on human HeLa cells. Specific trypsiligase recognition of the site of linkage was mediated by the dipeptide sequence Arg-His added to the proteins’ native N termini, pointing outside the cell membrane. The labeling procedure takes only about 5 minutes, as demonstrated for couplings of the fluorescence dye tetramethyl rhodamine and the affinity label biotin as well.     

Fabrication and joining mechanism of Nano-Cu/Si3N4 ceramic substrates

A novel method for fabricating a nano-Cu/Si₃N₄ ceramic substrate is proposed. The nano-Cu/Si₃N₄ ceramic substrate is first fabricated using spark plasma sintering (SPS) with the addition of nanoscale multilayer films (Ti/TiN/Ti/TiN/Ti) as transition layers. The microstructures of the nano-Cu metal layer and the interface between Cu and Si₃N₄ are investigated. The results show that a higher SPS temperature increases the grain size of the nano-Cu metal layer and affects the hardness. The microstructure of the transition layer evolves significantly after SPS. Ti in the transition layer can react with Si₃N₄ and with nano-Cu to form interfacial reaction layers of TiN and Ti–Cu, respectively; these ensure stronger bonding between nano-Cu and Si₃N₄. Higher SPS temperatures improve the diffusion ability of Ti and Cu, inducing the formation of Ti₃Cu₃O compounds in the nano-Cu metal layer and Ti₂Cu in the transition layer. This study provides an important strategy for designing and constructing a new type of ceramic substrate.     

Evaluation of standard and real fire exposures on thermal response of rail car floor assembly

This work developed a computational methodology to evaluate and compare standard fire exposures such as those outlined in ASTM E119 with real fire exposures and determine the difference in the temperature rise of a rail car floor assembly. The real fire exposures simulated in this work were identified in a review of incidents and consisted of a constantly-fed diesel fuel spill, a localized trash fire, and a gasoline spill simulated from a collision of the railcar with an automobile. These realistic fire exposures were applied to a variety of exemplar rail cars representative of single-level and bi-level passenger cars. These floor assembly models exposed to realistic fires were simulated in Fire Dynamics Simulator (FDS). The thermal exposure at the underside of railcar provided by FDS was coupled with a thermal model in ABAQUS, which provided the evolution of temperature in different components of the floor assembly. The standard scenarios were simulated for 2 hours instead of the typical 30 minutes to identify the appropriate exposure duration in ASTM E119, which can better represent a real fire scenario. The average and maximum temperatures predicted at the unexposed surface for both scenarios were compared with the threshold values given in NFPA 130.     

Design and Characterization of a New pVII Combinatorial Phage Display Peptide Library for Protease Substrate Mining Using Factor VII Activating Protease (FSAP) as Model

We describe a novel, easy and efficient combinatorial phage display peptide substrate-mining method to map the substrate specificity of proteases. The peptide library is displayed on the pVII capsid of the M13 bacteriophage, which renders pIII necessary for infectivity and efficient retrieval, in an unmodified state. As capture module, the 3XFLAG was chosen due to its very high binding efficiency to anti-FLAG mAbs and its independency of any post-translational modification. This library was tested with Factor-VII activating protease (WT-FSAP) and its single-nucleotide polymorphism variant Marburg-I (MI)-FSAP. The WT-FSAP results confirmed the previously reported Arg/Lys centered FSAP cleavage site consensus as dominant, as well as reinforcing MI-FSAP as a loss-of-function mutant. Surprisingly, rare substrate clones devoid of basic amino acids were also identified. Indeed one of these peptides was cleaved as free peptide, thus suggesting a broader range of WT-FSAP substrates than previously anticipated.     

二氧化锡溅射工艺研究

SnO2 是最早使用也是目前使用最广泛的一种气敏材料 ,使用该材料设计制作的气敏传感器具有许多优点。在简要介绍溅射镀膜的成膜过程和特点的基础上 ,着重介绍了SnO2 膜的制备流程 ,分析了功率和温度变化对成膜质量的影响     

地源热泵——辐射地板联供试验

通过地源热泵－辐射地板系统冬夏联供试验，讨论了该系统作为新型环保节能空调方式的可行性及优越性，并对运行中出现的问题进行分析。     

彩色PDP用玻璃基板

本文讨论了彩色ＰＤＰ现用钠钙玻璃的热稳定性，指出由于这种玻璃在热处理过程中易变形和收缩，因而不适合作大面积彩色ＰＤＰ基板材料，介绍了彩色ＰＤＰ基板玻璃的制造方法和特点。最后介绍了日本旭硝子公司和美国康宁公司各自为彩色ＰＤＰ新开发的玻璃基板材料ＰＤ２００和ＣＳ２５。     

无粘结预应力楼板新规范设计方法

目前国内现浇预应力楼板几乎均采用无粘结预应力方案,因而掌握其设计方法无疑具有现实意义。本文根据新规范,系统地介绍了高层建筑无粘结预应力楼板的设计方法,内容包括:设计原理、基本步骤及设计中需要注意的一些问题。     

环保中/高密度纤维板生产成本与性能问题 系统解决方案的研究(Ⅲ)

使用永港·伟方UF-1改性剂制备的改性脲醛胶粘剂生产E1级中纤板,胶粘剂生产成本降低10%,板材的各项力学性能均能达到国家标准,甲醛释放量达到了E1级标准,具有推广价值。     

晶体硅薄膜电池制备技术及研究现状

晶体硅薄膜太阳电池近些年来得到广泛的研究和初步的商业化探索。根据所采用的晶体硅薄膜沉积工艺中温度范围的不同，晶体硅薄膜电池研究可分为高温路线和低温路线两个不同发展方向。本文分别从这两个方向综述了目前国外晶体硅薄膜电池制备技术的最新进展，最新实验室研究结果。报导了晶体硅薄膜电池商业化进展状况，指出了晶体硅薄膜电池实现产业化必须解决的问题。