A hydroxypropyl methylcellulose plaque assay for human respiratory syncytial virus

ウイルスの研究において感染性のあるウイルス粒子を正確に定量することは，病原性の評価のみならずウイルスに対する新規治療薬の効果を客観的に評価するために極めて重要な手技である．プラークアッセイ法はウイルス粒子を定量するために不可欠な技術であるが，Respiratory syncytial virus（RSV）において，プラークアッセイ法を安定して行うことはしばしば困難である．今回我々は，overlay material（充填化合物）が細胞増殖に与える影響を中心にRSVおよびhuman metapneumovirus（hMPV）におけるプラークアッセイ法の最適化の検討を行った

The whole blood transcriptional regulation landscape in 465 COVID-19 infected samples from Japan COVID-19 Task Force

「コロナ制圧タスクフォース」COVID-19患者由来の血液細胞における遺伝子発現の網羅的解析 --重症度に応じた遺伝子発現の変化には、ヒトゲノム配列の個人差が影響する--. 京都大学プレスリリース. 2022-08-23.Coronavirus disease 2019 (COVID-19) is a recently-emerged infectious disease that has caused millions of deaths, where comprehensive understanding of disease mechanisms is still unestablished. In particular, studies of gene expression dynamics and regulation landscape in COVID-19 infected individuals are limited. Here, we report on a thorough analysis of whole blood RNA-seq data from 465 genotyped samples from the Japan COVID-19 Task Force, including 359 severe and 106 non-severe COVID-19 cases. We discover 1169 putative causal expression quantitative trait loci (eQTLs) including 34 possible colocalizations with biobank fine-mapping results of hematopoietic traits in a Japanese population, 1549 putative causal splice QTLs (sQTLs; e.g. two independent sQTLs at TOR1AIP1), as well as biologically interpretable trans-eQTL examples (e.g., REST and STING1), all fine-mapped at single variant resolution. We perform differential gene expression analysis to elucidate 198 genes with increased expression in severe COVID-19 cases and enriched for innate immune-related functions. Finally, we evaluate the limited but non-zero effect of COVID-19 phenotype on eQTL discovery, and highlight the presence of COVID-19 severity-interaction eQTLs (ieQTLs; e.g., CLEC4C and MYBL2). Our study provides a comprehensive catalog of whole blood regulatory variants in Japanese, as well as a reference for transcriptional landscapes in response to COVID-19 infection

DOCK2 is involved in the host genetics and biology of severe COVID-19

「コロナ制圧タスクフォース」COVID-19疾患感受性遺伝子DOCK2の重症化機序を解明 --アジア最大のバイオレポジトリーでCOVID-19の治療標的を発見--. 京都大学プレスリリース. 2022-08-10.Identifying the host genetic factors underlying severe COVID-19 is an emerging challenge. Here we conducted a genome-wide association study (GWAS) involving 2, 393 cases of COVID-19 in a cohort of Japanese individuals collected during the initial waves of the pandemic, with 3, 289 unaffected controls. We identified a variant on chromosome 5 at 5q35 (rs60200309-A), close to the dedicator of cytokinesis 2 gene (DOCK2), which was associated with severe COVID-19 in patients less than 65 years of age. This risk allele was prevalent in East Asian individuals but rare in Europeans, highlighting the value of genome-wide association studies in non-European populations. RNA-sequencing analysis of 473 bulk peripheral blood samples identified decreased expression of DOCK2 associated with the risk allele in these younger patients. DOCK2 expression was suppressed in patients with severe cases of COVID-19. Single-cell RNA-sequencing analysis (n = 61 individuals) identified cell-type-specific downregulation of DOCK2 and a COVID-19-specific decreasing effect of the risk allele on DOCK2 expression in non-classical monocytes. Immunohistochemistry of lung specimens from patients with severe COVID-19 pneumonia showed suppressed DOCK2 expression. Moreover, inhibition of DOCK2 function with CPYPP increased the severity of pneumonia in a Syrian hamster model of SARS-CoV-2 infection, characterized by weight loss, lung oedema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 has an important role in the host immune response to SARS-CoV-2 infection and the development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target

Microscopic characterization of metal-carbon-hydrogen composites (metal5Li, Mg)

Li-C-H system, which can store about 5.0 mass% of rechargeable H2, has been reported as a promising hydrogen storage system by Ichikawa et al. [Appl. Phys. Lett. 86, 241914 (2005); Mater. Trans. 46, 1757 (2005)]. This system was investigated from the thermodynamic and structural viewpoints. However, hydrogen absorption/desorption mechanism and the state of hydrogen atoms absorbed in the composite have not been clarified yet. In order to find new or better hydrogen storage system, graphite powder and nano-structural graphite ball-milled under H2 and Ar atmosphere were prepared and milled with Li and Mg under Ar atmosphere in this study. Microstructural analysis for those samples by transmission electron microscope revealed that LiC6 and/or LiC12 were formed in Li-C-H system. On the other hand, MgC2 was found in Mg-C-H system ball-milled under H2 atmosphere, but not in the system ball-milled under Ar atmosphere. These results indicated that nano-structure in composites of nano-structural graphite is different from that of alkali (-earth) metal. For these reasons, metal-C-H system can be recognized to be a new family of hydrogen storage materials