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.     

多晶硅表面金属催化化学腐蚀法制绒研究现状

在多晶硅太阳能电池的生产过程中, 金刚线切割技术(Diamond wire sawn, DWS)具有切割速度快、精度高、原材料损耗少等优点, 受到了广泛关注。金刚线切割多晶硅表面形成的损伤层较浅, 与传统的酸腐蚀制绒技术无法匹配, 金属催化化学腐蚀法应运而生。金属催化化学腐蚀法制绒具有操作简单、结构可控且易形成高深宽比的绒面等优点, 具有广阔的应用前景。本文总结了不同类型的金属催化剂在制绒过程中的腐蚀机理及其形成的绒面结构, 深入分析和讨论了具有代表性的银、铜的单一及复合催化腐蚀过程及绒面结构和电池片性能。最后对金刚线切割多晶硅片表面的金属催化化学腐蚀法存在的问题进行了分析, 并展望了未来的研究方向。     

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.     

Mechanical Properties of Single-Layer Diamond Reinforced Poly(vinyl alcohol) Nanocomposites through Atomistic Simulation

Low-dimensional carbon nanostructures are ideal nanofillers to reinforce the mechanical performance of polymer nanocomposites due to their excellent mechanical properties. Through molecular dynamics simulations, the mechanical performance of poly(vinyl alchohol) (PVA) nanocomposites reinforced with a single-layer diamond – diamane is investigated. It is found the PVA/diamane exhibits similar interfacial strengths and pull-out characteristics with the PVA/bilayer-graphene counterpart. Specifically, when the nanofiller is fully embedded in the nanocomposite, it is unable to deform simultaneously with the PVA matrix due to the weak interfacial load transfer efficiency, thus the enhancement effect is not significant. In comparison, diamane can effectively promote the tensile properties of the nanocomposite when it has a laminated structure as it deforms simultaneously with the matrix. With this configuration, the interlayer sp³ bonds endows diamane with a much higher resistance under compression and shear tests, thus the nanocomposite can reach very high compressive and shear stress. Overall, enhancement on the mechanical interlocking at the interface as triggered by surface functionalization is only effective for the fully embedded nanofiller. This work provides a fundamental understanding of the mechanical properties of PVA nanocomposites reinforced by diamane, which can shed lights on the design and preparation of next generation high-performance nanocomposites.     

Investigation of fabrication of gas diffusion substrate for proton exchange membrane fuel cells

The gas diffusion substrate (GDS) is essential in the proton exchange membrane fuel cells. Its fabrication techniques affect the performance significantly and are worthy of investigation. In this study, a manufacturing process of the GDS is proposed to understand the formation process of GDS and promote its structure and performance more pertinently. Different states during the preparation process, raw carbon paper, pre-curing, curing, carbonation, and graphitization, are characterized and measured. Experimental and numerical methods are employed to determine the relationships between microstructure, transport, and mechanical performance variation with the fabricating processes. The results show that its porosity, average pore size, and effective diffusivity decrease first and increase after curing. These parameters after graphitization are lower than that of the carbon paper (CP). The electrical resistivity increases dramatically while pre-curing and decreases gradually after curing, carbonation, and graphitization, and it is much reduced after graphitization. Moreover, mechanical measurement results show that both the picks of tensile strength and flexural modulus occur after curing. Its tensile strength shows little change after graphitization compared to the initial paper's. In contrast, the flexural modulus is improved significantly.     

Chromaticity-tunable remote LuYAG: Ce phosphor-in-glass film on regular textured glass substrate for white light emitting diodes

All inorganic remote phosphor-in-glass film exhibits excellent properties in high power white light-emitting-diodes (WLEDs) thanks to their easy fabrication and thermal stability. Herein, fabrication of (Lu, Y)₃Al₅O₁₂: Ce³⁺ (LuYAG: Ce)phosphors embedded in borosilicate glass film by the conventional solid state reaction and spin coating technology has been reported. The introduction of Y³⁺ ions reduces the difference of relative growth rate along some directions in growth of LuYAG microparticles, yielding a finer grain with smooth edges. By adjusting the molar concentration of Y³⁺ ions in LuAG phosphor, a series of tunable broadband emission from green to yellow region is observed and maintains excellent thermal stability. Meanwhile, the decay curves of samples with different Y³⁺ are almost same. SEM images show that phosphor particles are homogenously distributed within the glass matrix and keep their original morphology, suggesting the phosphor-in-glass films were synthesized as expected. Finally, a simple WLEDs based on the films was constructed using the commercial blue chip. The correlated color temperature ranging from 4853K to 4627K and high color rendering index from 81.4–79.7 were obtained. Upon the different driving current, the chromaticity coordinates of as-fabricated film exhibit good light color stability. These results bring an inspiring insight to tune the luminescent performance for remote WLEDs.     

Failure mechanism of MCrAlY coating at the coating‐substrate interface under type I hot corrosion

MCrAlY coatings are widely used to provide protection of hot component in modern gas turbine engines against high‐temperature oxidation and hot corrosion. Coating‐substrate interface, where the substrate is only partially covered by the ?coatings, is vulnerable to the hot corrosion attack. The accelerated degradation at the coating‐substrate interface can cause fast spallation of the coating, leading to the early failure of the gas turbine components. In this paper, MCrAlY powder was deposited on IN792 disks by high‐velocity oxygen‐fuel spraying. The hot corrosion behavior of the coated sample was investigated using (0.8Na, 0.2K)₂SO ₄ salt deposition at 900°C in lab air. Results showed a minor attack in the coating center, however, an accelerated corrosion attack at the coating‐substrate interface. The fast growth of corrosion products from substrate caused large local volume expansions at the coating‐substrate interface, resulting in an early coating spallation.     

基于数据拼接的金刚石刀具刃口三维形貌表征技术

原子力显微镜(AFM)是获取金刚石刀具刃口纳米级三维形貌的重要手段,但其存在测量范围小,难以表征参与切削区域整段刃口形貌特征的问题。采用基于迭代最近点法的刃口AFM测量数据拼接算法,获得了由多组数据拼接得到的刃口纳米级三维形貌,并进行了切削实验验证。实验结果表明,采用的数据拼接方法可较好地用于表征刀具刃口三维形貌。     

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.