Whole Genome Sequencing of Influenza A and B Viruses With the MinION Sequencer in the Clinical Setting: A Pilot Study

Introduction: Whole genome sequencing (WGS) of influenza viruses is important for preparing vaccines and coping with newly emerging viruses. However, WGS is difficult to perform using conventional next-generation sequencers in developing countries, where facilities are often inadequate. In this study, we developed a high-throughput WGS method for influenza viruses in clinical specimens with the MinION portable sequencer.Methods: Whole genomes of influenza A and B viruses were amplified by multiplex RT-PCR from 13 clinical specimens collected in Tokyo, Japan. Barcode tags for multiplex MinION sequencing were added with each multiplex RT-PCR amplicon by nested PCR with custom barcoded primers. All barcoded amplicons were mixed and multiplex sequencing using the MinION sequencer with 1D2 sequencing kit. In addition, multiplex RT-PCR amplicons generated from each clinical specimen were sequenced using the Illumina MiSeq platform to validate the performance of MinION sequencer. The accuracy, recall, and precision rates of MinION sequencing were calculated by comparing the results of variant calling in the Illumina MiSeq platform and MinION sequencer.Results: Whole genomes of influenza A and B viruses were successfully amplified by multiplex RT-PCR from 13 clinical samples. We identified 6 samples as influenza type A virus H3N2 subtype and 7 as influenza B virus Yamagata lineage using the Illumina MiSeq platform. The overall accuracy, recall, and precision rates of the MinION sequencer were, respectively 99.95%, 89.41%, and 97.88% from 1D reads and 99.97%, 93.28%, and 99.86% from 1D2 reads.Conclusion: We developed a novel WGS method for influenza A and B viruses. It is necessary to improve read accuracy and analytical tools in order to better utilize the MinION sequencer for real-time monitoring of genetic rearrangements and for evaluation of newly emerging viruses

Increased Stathmin1 Expression in the Dentate Gyrus of Mice Causes Abnormal Axonal Arborizations

Pituitary adenylate cyclase-activating polypeptide (PACAP) is involved in multiple brain functions. To clarify the cause of abnormal behavior in PACAP deficient-mice, we attempted the identification of genes whose expression was altered in the dentate gyrus of PACAP-deficient mice using the differential display method. Expression of stathmin1 was up-regulated in the dentate gyrus at both the mRNA and protein levels. PACAP stimulation inhibited stathmin1 expression in PC12 cells, while increased stathmin1expression in neurons of the subgranular zone and in primary cultured hippocampal neurons induced abnormal arborization of axons. We also investigated the pathways involved in PACAP deficiency. Ascl1 binds to E10 box of the stathmin1 promoter and increases stathmin1 expression. Inhibitory bHLH proteins (Hes1 and Id3) were rapidly up-regulated by PACAP stimulation, and Hes1 could suppress Ascl1 expression and Id3 could inhibit Ascl1 signaling. We also detected an increase of stathmin1 expression in the brains of schizophrenic patients. These results suggest that up-regulation of stathmin1 in the dentate gyrus, secondary to PACAP deficiency, may create abnormal neuronal circuits that cause abnormal behavior

Microstructure and analysis of oxide scales formed on Cr-Si-Ni compacts in air and H2O-containing atmosphere

The purpose of this study was to characterize the oxide scales formed on various Cr-Si-Ni compacts at 1273 K in air and H2O-containing atmosphere by TEM. It was found that CrSi2-(5-20) mass%Ni compacts form double layer scales, consisting of an outer Cr2O3 layer and an inner SiO2 layer. The oxide scale changed from SiO2- to Cr2O3-based scale with an increase in the Ni concentration. However, it was observed that the oxide scale formed in H2O-containing atmospheres showed local SiO2 growth into the substrate. This result suggests that the inward oxidant diffusion promotes the local growth of SiO2 in the H2O-containing atmospheres

Characteristics of Problem Consciousness of Indonesian Returnee Nurses Who Experienced Intercultural Exchange in Foreign Countries

The purpose of this study was to reveal the problems recognized by Indonesian nurses after returning to work in Indonesian health and medical facilities after work experience in nursing or nursing care work in Japan. The Root Cause Analysis (RCA) with the theme of "Issues of my own workplace" was conducted on three people including two Indonesian nurses and one Japanese nurse as a facilitator. The issue tree as the result of RCA was an analysis subject. As a result, a series of eight events emerged in the issue tree. Four root causes were extracted. Problem consciousness emerged when subjects became culturally aware of the difference in standards between their home country and Japan which was then identified as cultural awareness

Moralization of Japanese cultural norms among student sojourners in Japan

This study investigated the relationship between different domains of cultural adaptation among international students in Japan and the moralization of culture-specific norms. Newcomers may adapt certain norms of the host culture and ascribe moral meanings to initially nonmoral activities or objects. Building on the existing model of sociocultural adaptation, we investigated how different types of sociocultural adaptation are associated with the moralization of Japanese cultural norms. For international students in Japan, there are three aspects of sociocultural adaptation: academic, daily living, and interpersonal. Our results showed that cultural adaptation in the interpersonal domain, but not in the academic and daily living domains, predicted harsher moral judgments of behaviors that violated Japanese cultural norms. These findings suggest that international students who are well adjusted in the interpersonal domain gain an understanding of what is sanctioned in the Japanese cultural context and come to see certain behaviors as morally appropriate. We discuss several implications for further investigating the moralization of certain behaviors within the context of acculturation