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.     

Simulation studies on a rotating ring disk electrode: Effects of ionic migration with kinetic complication-disk results

In continuation to my previous work (Guha S. AIChE J. 2013;59(4):1390-1399), in this work, effects of ionic migration are evaluated for disk region of a rotating ring disk electrode system by numerically solving complex differential equations, developed for mass transfer along with kinetic complication in presence of ionic migration under limiting current condition. The system for simulation is 0.01 M Fe₂(SO₄)₃ solution with H₂SO₄ as supporting electrolyte. Simulation cases are presence and absence of ionic migration with kinetic complication (oxidation of Fe²⁺ to Fe³⁺ under O₂ pressure). Results show that concentration boundary layer thickness of reactant Fe³⁺ reduces appreciably and steady-state disk current reduces substantially in presence of migration. Simulated steady-state disk current in absence of migration case agrees well with published data. Results indicate higher Fe²⁺ concentration in presence of migration and thereby higher rate of oxidation of Fe²⁺ to Fe³⁺ at all rate constant values.     

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     

Electrochemical mass transfer between an impinging jet and a rotating disk in a confined system

This paper presents the mass transfer results from an impinging liquid jet to a rotating disk. The mass transfer coefficients were measured using the electrochemical limiting diffusion current technique (ELDCT). Rotational Reynolds number (Re_r) in the range of 3.4 × 10⁴–1.2 × 10⁵, jet Reynolds number (Re_j) 1.7 × 10⁴–5.3 × 10⁴ and non-dimensional jet-to-disk spacing (H/d) 2–8 were taken into consideration as parameters. It was found that the jet impingement resulted in a substantial enhancement in the mass transfer compared to the case of the rotating disk without jet.     

Midplane-symmetry breaking in the flow between two counter-rotating disks

This paper considers the axisymmetric steady flow driven by exact counter rotation of two co-axial disks of finite radius. At the edges of the rotating disks one of three conditions is (typically) imposed: (i) zero velocity, corresponding to a stationary, impermeable, cylindrical shroud (ii) zero normal velocity and zero tangential fluid traction, corresponding to a (confined) free surface and (iii) an edge constraint that is consistent with a similarity solution of von Kármán form. The similarity solution is valid in an infinite geometry and possesses a pitchfork bifurcation that breaks the midplane symmetry at a critical Reynolds number. In this paper, similar bifurcations of the global (finite-domain) flow are sought and comparisons are made between the resulting bifurcation structure and that found for the similarity solution. The aim is to assess the validity of the nonlinear similarity solutions in finite domains and to explore the sensitivity of the solution structure to edge conditions that are implicitly neglected when assuming a self-similar flow. It is found that, whilst the symmetric similarity solution can be quantitatively useful for a range of boundary conditions, the bifurcated structure of the finite-domain flow is rather different for each boundary condition and bears little resemblance to the self-similar flow.     

改善磁盘阵列性能的方法

磁盘阵列是解决计算机Ｉ／Ｏ瓶颈问题的有效方法之一，通过对现有磁盘阵列结构的研究，提出了４种改善磁盘阵列性能的方法，即良分利用磁盘带宽，平衡多盘的负载，减少奇偶检验数据存取时间和磁盘阵列Ｃａｃｈｅ技术等，分析结果表明：在事务处理应用领域，利用Ｃａｃｈｅ来把小写转化为大写可大大改善目前一般情况下，以一道数据大小来作为磁盘的基本存取大小是合适的选择，磁盘阵列负载平衡设计不足是指正常模式下系统请求在多盘间     

凸轮机构基圆半径的确定方法及其分析

基圆半径的确定方法，除了用图解法和试算法外，还可用解析法和图解分析法求解。４种方法各有优缺点，设计时要根据具体情况选择。     

某型杀伤枪榴弹引信弹道炸原因分析及再设计思考

通过对某型杀伤枪榴弹引信所发生的三次弹道炸的原因进行分析，提出用延期索替代延期药盘，并对有关机构进行再设计的对策。