Automatic polymerase chain reaction product detection system for food safety monitoring using zinc finger protein fused to luciferase

An automatic polymerase chain reaction (PCR) product detection system for food safety monitoring using zinc finger (ZF) protein fused to luciferase was developed. ZF protein fused to luciferase specifically binds to target double stranded DNA sequence and has luciferase enzymatic activity. Therefore, PCR products that comprise ZF protein recognition sequence can be detected by measuring the luciferase activity of the fusion protein. We previously reported that PCR products from Legionella pneumophila and Escherichia coli (E. coli) O157 genomic DNA were detected by Zif268, a natural ZF protein, fused to luciferase. In this study, Zif268–luciferase was applied to detect the presence of Salmonella and coliforms. Moreover, an artificial zinc finger protein (B2) fused to luciferase was constructed for a Norovirus detection system. In the luciferase activity detection assay, several bound/free separation process is required. Therefore, an analyzer that automatically performed the bound/free separation process was developed to detect PCR products using the ZF–luciferase fusion protein. By means of the automatic analyzer with ZF–luciferase fusion protein, target pathogenic genomes were specifically detected in the presence of other pathogenic genomes. Moreover, we succeeded in the detection of 10 copies of E. coli BL21 without extraction of genomic DNA by the automatic analyzer and E. coli was detected with a logarithmic dependency in the range of 1.0 × 10 to 1.0 × 10⁶ copies.     

Phosphorothioate Modification of mRNA Accelerates the Rate of Translation Initiation to Provide More Efficient Protein Synthesis

Messenger RNAs (mRNAs) with phosphorothioate modification (PS‐mRNA) to the phosphate site of A, G, C, and U with all 16 possible combinations were prepared, and the translation reaction was evaluated using an E. coli cell‐free translation system. Protein synthesis from PS‐mRNA increased in 12 of 15 patterns when compared with that of unmodified mRNA. The protein yield increased 22‐fold when the phosphorothioate modification at A/C sites was introduced into the region from the 5′‐end to the initiation codon. Single‐turnover analysis of PS‐mRNA translation showed that phosphorothioate modification increases the number of translating ribosomes, thus suggesting that the rate of translation initiation (rate of ribosome complex formation) is positively affected by the modification. The method provides a new strategy for improving translation by using non‐natural mRNA.     

Fluorinated Phenanthrenes as Aryne Precursors: PAH Synthesis Based on Domino Ring Assembly Using 1,1‐Difluoroallenes

On treatment with the catalyst InBr₃, 1,1‐difluoroallenes that bear a cyclopentene moiety and an aryl group underwent domino ring assembly in the presence or absence of N‐bromosuccinimide or N‐iodosuccinimide to afford aryne precursors such as three‐ringed ortho‐fluoro(halo)phenanthrenes, four‐ringed ortho‐fluoro(halo)tetraphenes, ortho‐fluoro(halo)chrysenes and fluoro[4]helicenes. Metalation of the aryne precursors followed by elimination of the fluoride resulted in the unprecedented systematic generation of arynes bearing π‐extended systems. Diels?Alder reactions of these arynes with isobenzofurans afforded the corresponding cycloadducts whose reductive aromatisation in an SnCl₂/HBr system furnished fully aromatised benzotriphenylenes. In addition, oxidative aryl?aryl coupling (the Scholl reaction) of these benzotriphenylenes facilitated the synthesis of ‘half HBCs’ (hexabenzocoronenes).     

On the first homology of automorphism groups of manifolds with geometric structures

Hermann and Thurston proved that the group of diffeomorphisms with compact support of a smooth manifold M which are isotopic to the identity is a perfect group. We consider the case where M has a geometric structure. In this paper we shall survey on the recent results of the first homology of the diffeomorphism groups which preserve a smooth G-action or a foliated structure on M. We also work in Lipschitz category. This research was partially supported by Grant-in-Aid for Scientific Research (No. 16540058), Japan Society for the Promotion of Science. This research was partially supported by Grant-in-Aid for Scientific Research (No. 14540093), Japan Society for the Promotion of Science.     

Effect of the Propionylation Method on the Deformability under Thermal Pressure of Block-Shaped Wood

Converting wood waste into thermoplastic materials is an attractive means of increasing its utilization because complex three-dimensional molded products can easily be obtained by press molding wood with thermoplasticity. Chemical modification, especially esterification, is a promising method for imparting thermoplasticity to wood. In this study, we produced multiple propionylated wood specimens using several propionylation methods and elucidated the factors affecting the deformability of the wood. Regardless of the method, all of the propionylated wood samples showed deformability in the tangential direction. However, in the longitudinal direction, not only the degree of propionylation but also the propionylation method had a significant influence on the deformability. The flow in the tangential direction occurred under a relatively low pressure, whereas the flow in the longitudinal direction occurred under higher pressure. The chemical composition and motility of each sample were evaluated using solid-state NMR measurements. With some propionylation methods, decomposition of the cellulose main chain occurred during the reaction, which had a dominant effect on the deformability of the wood in the longitudinal direction. These results indicate that the deformability of wood can be controlled by the appropriate selection of a propionylation method and its treatment conditions.     

The line broadening and shift effects on the CO J=5-4 transition at 576 GHz induced by collisions with rare gases

The line profile of the CO J=5-4 transition at 576 GHz was measured precisely by a BWO-based submillimeter wave spectrometer. The foreign-gas effect on the broadening and shift of rare gases were determined together with the self-collision effects. The observed profiles exhibit clearly the Dicke narrowing effect for the cases of rare gas mixtures, and the Galatry function was used as the lineshape function in the analysis.     

Simplified technique of fabricating dielectric-coated silver hollow nickel waveguides by the outer-coating method of the liquid phase process

Considerable simplification has been made in the fabrication of rugged dielectric-coated silver hollow nickel waveguides. The new method is based on the outer-coating method and does not use any expensive equipment; such as RF-sputtering apparatus. In order to show its effectiveness; polyimide-coated silver hollow nickel-waveguides have been successfully fabricated with small attenuation.     

Investigating the competitive assumption of Multinomial Logit models of brand choice by nonparametric modeling

Summary  The Multinomial Logit (MNL) model is still the only viable option to study nonlinear responsiveness of utility to covariates nonparametrically. This research investigates whether MNL structure of inter-brand competition is a reasonable assumption, so that when the utility function is estimated nonparametrically, the IIA assumption does not bias the result. For this purpose, the authors compare the performance of two comparable nonparametric choice models that differ in one aspect: one assumes MNL competitive structure and the other infers the pattern of brands? competition nonparametrically from data.