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.     

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.     

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.     

Thermal stability enhancement of Cr2AlC coatings on Zr by utilizing a double layer diffusion barrier

Decomposition of Cr₂AlC deposited onto a Zr substrate and vacuum-annealed is observed at 800 °C as Al diffuses from the MAX phase into the Zr substrate. A double layer of ZrN and AlN has been predicted by CALPHAD calculations to act as diffusion barrier between the Zr substrate and Cr₂AlC. Experimental thermal stability investigations corroborate this prediction by confirming that the proposed double layer diffusion barrier coatings suppress the decomposition of Cr₂AlC for one hour at temperatures of up to 1000 °C.     

Polylactic acid as a biodegradable material for all-solution-processed organic electronic devices

In this paper we report on the fabrication of spin-coated biodegradable polylactic acid (PLA) thin films to be used as substrates for the realisation of all-solution-processed organic electronic devices. The full mechanical and electrical characterisation of these substrates shows that they exhibit good mechanical and dielectric properties and are therefore suitable for the fabrication of disposable electronics. To demonstrate practically the functionality of such PLA thin films, organic electronic devices were realised on the top of them, exclusively by means of solution-process fabrication techniques and in particular inkjet-printing. Also, a photonic curing procedure is here presented as a means for sintering the conductive inks without heating up the PLA substrates. Two types of organic transistors were fabricated on the top of PLA: organic field-effect transistors (OFETs), where the PLA film was used not only as a substrate but also as the gate dielectric, and all-inkjet-printed organic electrochemical transistors (OECTs). The second typology of transistors exhibited one of the highest transconductance reported so far in the literature (up to 2.75 mS). This study opens an avenue for the fabrication of disposable, low-cost organic electronic devices.     

The impact of inoculum source,inoculum to substrate ratio and sample preservation on methane potential from different substrates

Batch experiments were conducted to evaluate the impact of inoculum sources, inoculum to substrate (IS) ratio and storage conditions on the potential and production rate of methane (CH₄) from different substrates: wheat straw, whole crop maize, cattle manure, grass and cellulose.The results of the test with four inocula and four substrates indicated that inoculum source could have a significant impact on both CH₄ potential (BMP) and the kinetics parameters of different substrates. The two inocula showing the highest BMP and production rates in each period were those coming from a feeding with more than 70% of animal manure under thermophilic conditions. The impact of the IS ratio in the range 0.25–2.5, in terms of g volatile solids (VS) substrate/g VS inoculum, depended on substrate type. Maize silage was more affected to changes in the IS ratio than wheat straw. The optimal IS ratio range for maize was 1.0–1.5, however, a wider IS range can be used in wheat straw (0.5–2.5). The impact of freezing and drying depended on biomass type. Freezing, drying and ensiling of grass increased the CH₄ yield compared to fresh grass. Drying of maize had no impact while freezing reduced the CH₄ potential. Drying and freezing had no impact on straw.