Study of Complete Genome Sequences of Rotavirus A Epidemics and Evolution in Japan in 2012–2014

A comprehensive molecular epidemiological study using next-generation sequencing technology was conducted on 333 rotavirus A (RVA)-positive specimens collected from six sentinel hospitals across Japan over three consecutive seasons (2012–2014). The majority of the RVA isolates were grouped into five genotype constellations: Wa-like G1P[8], DS-1-like G1P[8], G2P[4], G3P[8] and G9P[8]. Phylogenetic analysis showed that the distribution of strains varied by geographical locations and epidemic seasons. The VP7 genes of different G types were estimated to evolve at 7.26 × 10-4–1.04 × 10-3 nucleotide substitutions per site per year. The Bayesian time-scaled tree of VP7 showed that the time to the most recent common ancestor of epidemic strains within a region was 1–3 years, whereas that of the epidemic strains across the country was 2–6 years. This study provided, for the first time, the timeframe during which an epidemic strain spread locally and within the country and baseline information needed to predict how rapidly RVAs spread

Mutational study of sapovirus expression in insect cells

Human sapovirus (SaV), an agent of human gastroenteritis, cannot be grown in cell culture, but expression of the recombinant capsid protein (rVP1) in a baculovirus expression system results in the formation of virus-like particles (VLPs). In this study we compared the time-course expression of two different SaV rVP1 constructs. One construct had the native sequence (Wt construct), whereas the other had two nucleotide point mutations in which one mutation caused an amino acid substitution and one was silent (MEG-1076 construct). While both constructs formed VLPs morphologically similar to native SaV, Northern blot analysis indicated that the MEG-1076 rVP1 mRNA had increased steady-state levels. Furthermore, Western blot analysis and an antigen enzyme-linked immunosorbent assay showed that the MEG-1076 construct had increased expression levels of rVP1 and yields of VLPs. Interestingly, the position of the mutated residue was strictly conserved residue among other human SaV strains, suggesting an important role for rVP1 expression

Crystalline maricite NaFePO₄ as a positive electrode material for sodium secondary batteries operating at intermediate temperature

Maricite NaFePO₄ (m-NaFePO₄) was investigated as a positive electrode material for intermediate-temperature operation of sodium secondary batteries using ionic liquid electrolytes. Powdered m-NaFePO₄ was prepared by a conventional solid-state method at 873 K and subsequently fabricated in two different conditions; one is ball-milled in acetone and the other is re-calcined at 873 K after the ball-milling. Electrochemical properties of the electrodes prepared with the as-synthesized m-NaFePO₄, the ball-milled m-NaFePO₄, and the re-calcined m-NaFePO₄ were investigated in Na[FSA]-[C₂C₁im][FSA] (C₂C₁im⁺ = 1-ethyl-3-methylimidazolium, FSA⁻ = bis(fluorosulfonyl)amide) ionic liquid electrolytes at 298 K and 363 K to assess the effects of temperature and particle size on their electrochemical properties. A reversible charge-discharge capacity of 107 mAh g⁻¹ was achieved with a coulombic efficiency >98% from the 2nd cycle using the ball-milled m-NaFePO₄ electrode at a C–rate of 0.1 C and 363 K. Electrochemical impedance spectroscopy using m-NaFePO₄/m-NaFePO₄ symmetric cells indicated that inactive m-NaFePO₄ becomes an active material through ball-milling treatment and elevation of operating temperature. X-ray diffraction analysis of crystalline m-NaFePO₄ confirmed the lattice contraction and expansion upon charging and discharging, respectively. These results indicate that the desodiation-sodiation process in m-NaFePO₄ is reversible in the intermediate-temperature range

Inhibition of Cellular Protein Secretion by Norwalk Virus Nonstructural Protein p22 Requires a Mimic of an Endoplasmic Reticulum Export Signal

Protein trafficking between the endoplasmic reticulum (ER) and Golgi apparatus is central to cellular homeostasis. ER export signals are utilized by a subset of proteins to rapidly exit the ER by direct uptake into COPII vesicles for transport to the Golgi. Norwalk virus nonstructural protein p22 contains a YXΦESDG motif that mimics a di-acidic ER export signal in both sequence and function. However, unlike normal ER export signals, the ER export signal mimic of p22 is necessary for apparent inhibition of normal COPII vesicle trafficking, which leads to Golgi disassembly and antagonism of Golgi-dependent cellular protein secretion. This is the first reported function for p22. Disassembly of the Golgi apparatus was also observed in cells replicating Norwalk virus, which may contribute to pathogenesis by interfering with cellular processes that are dependent on an intact secretory pathway. These results indicate that the ER export signal mimic is critical to the antagonistic function of p22, shown herein to be a novel antagonist of ER/Golgi trafficking. This unique and well-conserved human norovirus motif is therefore an appealing target for antiviral drug development

Genetic Diversity of Sapovirus in Children, Australia

Sapovirus was detected in 7 of 95 stool specimens from children with gastroenteritis of unknown etiology in Sydney, Australia, from August 2001 to August 2002 and from February 2004 to August 2004, by using reverse transcription–polymerase chain reaction. Sequence analysis of the N-terminal capsid region showed all human sapovirus genogroups

Degradation of Organic Coatings on Steel Investigated by Dynamic Electrochemical Impedance Spectroscopy

ABSTRACT Degradation of the organic coatings on steel has been investigated by the dynamic impedance measurements. The electrochemical impedance spectroscopy (EIS) has been used to evaluate the corrosion of steel coated by the organic film. Generally the electrochemical impedance of the coated steel shows complicated behavior, because the degradation of organic coating involves various steps until the steel corrodes under the coating. In the present paper, time variation of electrochemical impedance was measured to investigate the initial stage of coating degradation. The dynamic impedance was displayed on 3-dimension plots, whose axes were real and imaginary components and time. The time variation of the film resistance related to the permeation of water was monitored to discuss the corrosion mechanisms of coated steel

Synthesis of carbon nanotubes by microwave heating: Influence of diameter of catalytic Ni nanoparticles on diameter of CNTs

We rapidly synthesized multi walled carbon nanotubes (MWCNTs) by calcination of granulated polystyrene with nickel nanoparticles having different average diameter (D-Ni = 10, 20, 50 or 90 nm) under nitrogen gas at a certain temperature and time (700 degrees C, 15 min or 800 degrees C, 10 min), using a domestic microwave oven in order to systematically investigate the influence of the diameter of nickel nanoparticles on the diameter of MWCNTs. The MWCNTs synthesized here were characterized by a transmission electron microscope, a Raman spectrophotometer and a wide angle X-ray diffractometer. We found that for the calcination condition of (800 degrees C, 10 min), a relationship between the outer diameter of the resulted carbon nanotubes (D-CNT) and the diameter of catalytic nickel nanoparticles (D-Ni) can be described as a linear function, D-CNT = 1.01D(Ni) + 14.79 nm with the correlation coefficient R = 0.99, and that for the calcination condition of 700 degrees C, 15 min, D-CNT = 1.12D(Ni) + 7.80 nm with R = 0.95. Thus, we revealed that when the diameter of the catalytic nickel nanoparticles (D-Ni) increases by 1 nm, the outer diameter of the obtained MWCNTs (D-CNT) increases by about 1 nm.ArticleJOURNAL OF MATERIALS CHEMISTRY A. 2(8):2773-2780 (2014)journal articl

Improvement of Rotavirus Genotyping Method by Using the Semi-Nested Multiplex-PCR With New Primer Set

Rotavirus A (RVA) is a major cause of gastroenteritis in infants and young children. After vaccine introduction, RVA surveillance has become more important for monitoring changes in genotype distribution, and the semi-nested multiplex-PCR is a popular method for RVA genotyping. In particular, the VP7 primer set reported by Gouvea and colleagues in 1990 is still widely used worldwide as the recommended WHO primer set in regional and national reference RVA surveillance laboratories. However, this primer set yielded some mistakes with recent epidemic strains. The newly emerged equine-like G3 strains were mistyped as G1, G8 strains were mistyped as G3, the G9 lineage 3 strains showed very weak band, and the G9 lineage 6 strains showed a G9-specific band and a non-specific band. Gouvea’s standard protocol has become relatively unreliable for identifying genotypes correctly. To overcome this limitation, we redesigned the primer set to include recent epidemic strains. Our new primer set enabled us to correctly identify the VP7 genotypes of representative epidemic strains by agarose gel electrophoresis (G1, G2, human typical G3, equine-like G3, G4, G8, G9, and G12). We believe that the multiplex-PCR method with our new primer set is a useful and valuable tool for surveillance of RVA epidemics

Predicting Directions of Changes in Genotype Proportions Between Norovirus Seasons in Japan

The norovirus forecasting system (NOROCAST) has been developed for predicting directions of changes in genotype proportions between human norovirus (HuNoV) seasons in Japan through modeling herd immunity to structural protein 1 (VP1). Here 404 nearly complete genomic sequences of HuNoV were analyzed to examine whether the performance of NOROCAST could be improved by modeling herd immunity to VP2 and non-structural proteins (NS) in addition to VP1. It was found that the applicability of NOROCAST may be extended by compensating for unavailable sequence data and observed genotype proportions of 0 in each season. Incorporation of herd immunity to VP2 and NS did not appear to improve the performance of NOROCAST, suggesting that VP1 may be a suitable target of vaccines