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.     

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.     

Food recognition using neural network classifier and multiple hypotheses image segmentation

ABSTRACT

This paper proposes the multiple-hypotheses image segmentation and feed-forward neural network classifier for food recognition to improve the performance. Initially, the food or meal image is given as input. Then, the segmentation is applied to identify the regions, where a particular food item is located using salient region detection, multi-scale segmentation, and fast rejection. Then, the features of every food item are extracted by the global feature and local feature extraction. After the features are obtained, the classification is performed for each segmented region using a feed-forward neural network model. Finally, the calorie value is computed with the aid of (i) food volume and (ii) calorie and nutrition measure based on mass value. The experimental results and performance evaluation are validated. The outcome of the proposed method attains 0.947 for Macro Average Accuracy (MAA) and 0.959 for Standard Accuracy (SA), which provides better classification performance.     

加工高酸油品加氢反应进料加热炉炉管的选材

论述了加氢装置反应进料加热炉炉管经常出现的晶间腐蚀的形成原因、控制方法以及近年来油品中出现的环烷酸对炉管的腐蚀影响,并结合工程实例,讨论了综合考虑强度和防止这两种腐蚀的炉管选材的问题。     

地震平衡技术及其在地震反演中的应用

地震平衡技术是一种在地震反演中改善井震关系的新技术。它通过单井对比、空间分形建模以及平衡处理的手段 ,克服地震反演中子波空变的影响 ,使得经过地震平衡技术处理的地震剖面井震关系得到较好改善 ,从而可以用统一的子波对研究区的地震资料进行反演     

Al、Mo含量对铸造钛合金力学性能的影响

运用正交实验 ,考察了Al、Mo含量对Ti Al Mo 1Zr系铸造钛合金力学性能的影响。试验结果表明 :随Al、Mo含量提高 ,铸造合金的强度增加 ,塑性和冲击韧性降低 ,但Al、Mo的交互作用却使合金塑性提高 ,强度和冲击韧性降低     

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     

Eine Definitionssprache für die Kommunikationsverbindungen in verteilten Automatisierungssystemen

Eingegangen am 13.11.1996, in überarbeiteter Form am 12.5.1997     

网络安全中的防火墙技术

介绍了防火墙的原理、分类及其应用