Sofosbuvir Selects for Drug-Resistant Amino Acid Variants in the Zika Virus RNA-Dependent RNA-Polymerase Complex In Vitro

The nucleotide analog sofosbuvir, licensed for the treatment of hepatitis C, recently revealed activity against the Zika virus (ZIKV) in vitro and in animal models. However, the ZIKV genetic barrier to sofosbuvir has not yet been characterized. In this study, in vitro selection experiments were performed in infected human hepatoma cell lines. Increasing drug pressure significantly delayed viral breakthrough (p = 0.029). A double mutant in the NS5 gene (V360L/V607I) emerged in 3 independent experiments at 40–80 µM sofosbuvir resulting in a 3.9 ± 0.9-fold half- maximal inhibitory concentration (IC₅₀) shift with respect to the wild type (WT) virus. A triple mutant (C269Y/V360L/V607I), detected in one experiment at 80 µM, conferred a 6.8-fold IC₅₀ shift with respect to the WT. Molecular dynamics simulations confirmed that the double mutant V360L/V607I impacts the binding mode of sofosbuvir, supporting its role in sofosbuvir resistance. Due to the distance from the catalytic site and to the lack of reliable structural data, the contribution of C269Y was not investigated in silico. By a combination of sequence analysis, phenotypic susceptibility testing, and molecular modeling, we characterized a double ZIKV NS5 mutant with decreased sofosbuvir susceptibility. These data add important information to the profile of sofosbuvir as a possible lead for anti-ZIKV drug development.     

影响煤层气开发效果的地质因素再认识

地质特征认识对煤层气开发效果起着重要作用。在资源特征相差不大的情况下,发现煤层气相邻井的产量差异仍较大。排除工程因素后,通过选取8类地质参数,细致比对了保德区块低产井与邻井的参数特征,筛查出其关键因素为煤层微幅构造与顶板封盖条件,其中以微幅构造为主。据此,重新认识并划分出区块新的次生褶皱背斜单元、向斜单元和斜坡单元,获得了不同次生褶皱构造单元的开发特征。结果表明,高、低产井分布与次生褶皱背斜、向斜相关性高达92%。其中:高产井主要分布在次生褶皱背斜变化较缓、呈隆起状的“平台”,且煤层顶板以泥岩、碳质泥岩为主,封盖性较好;低产井主要分布在次生褶皱向斜,同一井台各井开发效果差异表现为从向斜条带轴部—向斜条带内—向斜条带外的煤层气井平均单井产量不断增加,到向斜轴部的距离大于向斜曲率半径73.5%的范围为主力产气区,小于向斜曲率半径40.0%范围为产水主力区。这对煤层气新井部署、生产管理、开发调整等,具有一定的指导意义。     

Electrocatalytic Reduction of CO2 to Ethylene by Molecular Cu‐Complex Immobilized on Graphitized Mesoporous Carbon

The electrochemical reduction of carbon dioxide (CO₂) to hydrocarbons is a challenging task because of the issues in controlling the efficiency and selectivity of the products. Among the various transition metals, copper has attracted attention as it yields more reduced and C2 products even while using mononuclear copper center as catalysts. In addition, it is found that reversible formation of copper nanoparticle acts as the real catalytically active site for the conversion of CO₂ to reduced products. Here, it is demonstrated that the dinuclear molecular copper complex immobilized over graphitized mesoporous carbon can act as catalysts for the conversion of CO₂ to hydrocarbons (methane and ethylene) up to 60%. Interestingly, high selectivity toward C2 product (40% faradaic efficiency) is achieved by a molecular complex based hybrid material from CO₂ in 0.1 m KCl. In addition, the role of local pH, porous structure, and carbon support in limiting the mass transport to achieve the highly reduced products is demonstrated. Although the spectroscopic analysis of the catalysts exhibits molecular nature of the complex after 2 h bulk electrolysis, morphological study reveals that the newly generated copper cluster is the real active site during the catalytic reactions.     

Antitumor Effects of Vitamin D Analogs on Hamster and Mouse Melanoma Cell Lines in Relation to Melanin Pigmentation

Deregulated melanogenesis is involved in melanomagenesis and melanoma progression and resistance to therapy. Vitamin D analogs have anti-melanoma activity. While the hypercalcaemic effect of the active form of Vitamin D (1,25(OH)₂D₃) limits its therapeutic use, novel Vitamin D analogs with a modified side chain demonstrate low calcaemic activity. We therefore examined the effect of secosteroidal analogs, both classic (1,25(OH)₂D₃ and 25(OH)D₃), and novel relatively non-calcemic ones (20(OH)D₃, calcipotriol, 21(OH)pD, pD and 20(OH)pL), on proliferation, colony formation in monolayer and soft-agar, and mRNA and protein expression by melanoma cells. Murine B16-F10 and hamster Bomirski Ab cell lines were shown to be effective models to study how melanogenesis affects anti-melanoma treatment. Novel Vitamin D analogs with a short side-chain and lumisterol-like 20(OH)pL efficiently inhibited rodent melanoma growth. Moderate pigmentation sensitized rodent melanoma cells towards Vitamin D analogs, and altered expression of key genes involved in Vitamin D signaling, which was opposite to the effect on heavily pigmented cells. Interestingly, melanogenesis inhibited ligand-induced Vitamin D receptor translocation and ligand-induced expression of VDR and CYP24A1 genes. These findings indicate that melanogenesis can affect the anti-melanoma activity of Vitamin D analogs in a complex manner.     

A facile method for preparation of PGS@ZB‐N and gas‐sol alternating synergistic effect for fire resistance of EVA

Through the simple precipitation of palygorskite (PGS) by zinc borate (ZB) (to make PGS@ZB) and the decoration of PGS@ZB by dodecylamine (N), a novel organic‐inorganic@inorganic hybrid flame retardant of PGS@ZB‐N was prepared and was incorporated with ethylene vinyl acetate copolymer (EVA) to improve its flame retardance. The structure and morphology of PGS@ZB‐N were characterized by Fourier transform infrared (FTIR) spectroscopy, X‐ray diffraction (XRD), and scanning electron microscopy (SEM), and it was confirmed that the PGS@ZB‐N hybrid had been successfully prepared. The flame retardancy and burning behavior of EVA/PGS@ZB‐N/EG (EG = expandable graphite) composite were studied through thermogravimetric analysis (TGA), limiting oxygen index (LOI), UL‐94 (by the vertical burning test), and cone calorimeter test (CCT) characterizations. The prepared EVA/PGS@ZB‐N/EG composite obtained an LOI value of 41.2% with the addition of 30 wt% PGS@ZB‐N/EG. It was found that EVA/PGS@ZB‐N/EG was protected through a gas phase and condensed phase alternating synergistic effect mechanism.     

Genes Identification,Molecular Docking and Dynamics Simulation Analysis of Laccases from Amylostereum areolatum Provides Molecular Basis of Laccase Bound to Lignin

An obligate mutualistic relationship exists between the fungus Amylostereum areolatum and woodwasp Sirex noctilio. The fungus digests lignin in the host pine, providing essential nutrients for the growing woodwasp larvae. However, the functional properties of this symbiosis are poorly described. In this study, we identified, cloned, and characterized 14 laccase genes from A. areolatum. These genes encoded proteins of 508 to 529 amino acids and contained three typical copper-oxidase domains, necessary to confer laccase activity. Besides, we performed molecular docking and dynamics simulation of the laccase proteins in complex with lignin compounds (monomers, dimers, trimers, and tetramers). AaLac2, AaLac3, AaLac6, AaLac8, and AaLac10 were found that had low binding energies with all lignin model compounds tested and three of them could maintain stability when binding to these compounds. Among these complexes, amino acid residues ALA, GLN, LEU, PHE, PRO, and SER were commonly present. Our study reveals the molecular basis of A. areolatum laccases interacting with lignin, which is essential for understanding how the fungus provides nutrients to S. noctilio. These findings might also provide guidance for the control of S. noctilio by informing the design of enzyme mutants that could reduce the efficiency of lignin degradation.     

Sequence-Dependent Nanofiber Structures of Phenylalanine and Isoleucine Tripeptides

The molecular design of short peptides to achieve a tailor-made functional architecture has attracted attention during the past decade but remains challenging as a result of insufficient understanding of the relationship between peptide sequence and assembled supramolecular structures. We report a hybrid-resolution model to computationally explore the sequence–structure relationship of self-assembly for tripeptides containing only phenylalanine and isoleucine. We found that all these tripeptides have a tendency to assemble into nanofibers composed of laterally associated filaments. Molecular arrangements within the assemblies are diverse and vary depending on the sequences. This structural diversity originates from (1) distinct conformations of peptide building blocks that lead to different surface geometries of the filaments and (2) unique sidechain arrangements at the filament interfaces for each sequence. Many conformations are available for tripeptides in solution, but only an extended β-strand and another resembling a right-handed turn are observed in assemblies. It was found that the sequence dependence of these conformations and the packing of resulting filaments are determined by multiple competing noncovalent forces, with hydrophobic interactions involving Phe being particularly important. The sequence pattern for each type of assembly conformation and packing has been identified. These results highlight the importance of the interplay between conformation, molecular packing, and sequences for determining detailed nanostructures of peptides and provide a detailed insight to support a more precise design of peptide-based nanomaterials.     

Influence of the ZrB2 content on the anti-oxidation ability of ZrB2-SiC coatings in aerobic environments with broad temperature range

ZrB₂-SiC coatings with different ZrB₂ contents were prepared by liquid phase sintering. The oxidation processes of coatings were explained according to TG results of ZrB₂-SiC coatings and powders tested from 298 K to 1773 K. Results show that, increasing ZrB₂ content made the weight of the samples changed from weight-loss of 10.04% to weight-gain of 0.14%, while the fastest weight-loss regions were narrowed, whose inflection points reduced from 1310℃ to 1050℃. Increasing ZrB₂ content made the relative oxygen permeability of the ZrB₂-SiC/SiC coatings reduced from 40%–60% to -10%-5%. Increasing ZrB₂ content enhanced high-temperature stability of coatings, making final weight of samples changed from weight-loss of 0.16% to weight-gain of 0.11% after oxidation at 1773 K for 200 h. The peeling and dispersion of Zr-oxides formed Zr-B-Si-O compound glass layer, presenting enhanced stability, dispersion strengthening and pinning effect of Zr-oxides, which were responsible for the excellent anti-oxidation protective effect of coatings in a broad temperature region.     

“Size effect” in the fatigue behavior of Friction Stir Welded plates

Comparative fatigue tests were carried out on Friction Stir Welded specimens of a 2195-T8 aluminum–lithium alloy that differed significantly in width. The width of the larger specimens was over thirteen times greater than that of the small specimens. Fatigue results showed a clear “size effect”, i.e. fatigue life of large specimens was about 40% of the corresponding value of small specimens. The Equivalent Initial Flaw Size methodology was adopted to correlate the two sets of results. Fatigue crack initiation life was disregarded with respect to crack propagation life, and fatigue life was evaluated only as propagation of a small pre-existing defect. Following this methodology, test results of small specimens were used to evaluate the initial equivalent flaw contained in each specimen. It was assumed that this data followed a normal distribution. The equivalent initial flaw in larger specimens was evaluated by simple geometrical considerations. A very good assessment of mean fatigue life and scatter in the fatigue results of large specimens was obtained by simulating the propagation of these defects. Calculations were carried out by taking also welding residual stresses into account, but the results demonstrated that this effect was not significant.     

Facile construction of composition-tuned ruthenium-nickel nanoparticles on g-C3N4 for enhanced hydrolysis of ammonia borane without base additives

Developing high-efficiency and low-cost catalysts for hydrogen evolution from hydrolysis of ammonia borane (AB) is significant and critical for the exploitation and utilization of hydrogen energy. Herein, the in-situ fabrication of well-dispersed and small bimetallic RuNi alloy nanoparticles (NPs) with tuned compositions and concomitant hydrolysis of AB are successfully achieved by using graphitic carbon nitride (g-C₃N₄) as a NP support without additional stabilizing ligands. The optimized Ru₁Ni_7.5/g-C₃N₄ catalyst exhibits an excellent catalytic activity with a high turnover frequency of 901 min^?1 and an activation energy of 28.46 kJ mol^?1 without any base additives, overtaking the activities of many previously reported catalysts for AB hydrolysis. The kinetic studies indicate that the AB hydrolysis over Ru₁Ni_7.5/g-C₃N₄ is first-order and zero-order reactions with respect to the catalyst and AB concentrations, respectively. Ru₁Ni_7.5/g-C₃N₄ has a good recyclability with 46% of the initial catalytic activity retained even after five runs. The high performance of Ru₁Ni_7.5/g-C₃N₄ should be assigned to the small-sized alloy NPs with abundant accessible active sites and the synergistic effect between the composition-tuned Ru–Ni bimetals. This work highlights a potentially powerful and simple strategy for preparing highly active bimetallic alloy catalysts for AB hydrolysis to generate hydrogen.