Mutation Analysis of 2009 Pandemic Influenza A(H1N1) Viruses Collected in Japan during the Peak Phase of the Pandemic

BACKGROUND: Pandemic influenza A(H1N1) virus infection quickly circulated worldwide in 2009. In Japan, the first case was reported in May 2009, one month after its outbreak in Mexico. Thereafter, A(H1N1) infection spread widely throughout the country. It is of great importance to profile and understand the situation regarding viral mutations and their circulation in Japan to accumulate a knowledge base and to prepare clinical response platforms before a second pandemic (pdm) wave emerges. METHODOLOGY: A total of 253 swab samples were collected from patients with influenza-like illness in the Osaka, Tokyo, and Chiba areas both in May 2009 and between October 2009 and January 2010. We analyzed partial sequences of the hemagglutinin (HA) and neuraminidase (NA) genes of the 2009 pdm influenza virus in the collected clinical samples. By phylogenetic analysis, we identified major variants of the 2009 pdm influenza virus and critical mutations associated with severe cases, including drug-resistance mutations. RESULTS AND CONCLUSIONS: Our sequence analysis has revealed that both HA-S220T and NA-N248D are major non-synonymous mutations that clearly discriminate the 2009 pdm influenza viruses identified in the very early phase (May 2009) from those found in the peak phase (October 2009 to January 2010) in Japan. By phylogenetic analysis, we found 14 micro-clades within the viruses collected during the peak phase. Among them, 12 were new micro-clades, while two were previously reported. Oseltamivir resistance-related mutations, i.e., NA-H275Y and NA-N295S, were also detected in sporadic cases in Osaka and Tokyo

ダイ49ジ ナンキョク チイキ カンソクタイ ナツタイ ニオケル コショウ カンソク

第49次日本南極地域観測隊(第49次)夏隊において,湖沼観測として湖沼環境観測,生物・生態学的研究試料としての湖水と湖底の生物群集採取,及び現場実験を宗谷海岸露岩域にある複数の湖沼で実施した.この湖沼観測報告は南極観測事業第VII期計画の一般プロジェクト研究(P3)「極域環境変動と生態系変動に関する研究」及びモニタリング研究観測(M4)「生態系変動のモニタリング」の両課題にかかわる観測を記録したものである.野外観測は2007年12月22日から2008年2月13日の期間,砕氷船「しらせ」が昭和基地沖近傍に滞在中に実施した.今回は夏季の湖沼環境変動と湖底の生物(藻類群集)の応答を集中的に観測すべく,スカルブスネスの長池にて観測とサンプリング・現場実験を繰り返し実施する一方,きざはし浜生物観測小屋から徒歩日帰り圏内にある周辺の14湖沼,及びヘリコプターを利用した日帰り観測にてスカルブスネス東部の4湖沼,及び他の露岩,スカーレンにあるスカーレン大池,ラングホブデ域の雪鳥池・東雪鳥池,ぬるめ池にて湖沼水質環境観測と試料採集を適宜実施した.このうち,スカルブスネス東部のなまず池 (仮称)では潜水による水中設置ビデオ装置の回収と,湖底のコケ類・藻類が作り上げている「とさか・筍状」の群落の採集,ラングホブデぬるめ池では湖底から小型カイアシ類の定量サンプリングを実施,これらを研究試料として日本に持ち帰ることができた.また,第47次隊により雪の堤防の決壊の発見(第46次越冬期間中に決壊したとみられる)が報告されたラングホブデ南部の平頭氷河末端にあった「氷河池」(仮称)の現状視察も実施,決壊前後での3m以上と思われる大幅な水位変動痕からフィルム状の生物試料を採集し持ち帰った.Observations on the limnological properties, samplings of waters and bottom assemblages for biological and ecological studies, and some field experimental studies at several lakes in Soya Coast ice-free areas, were carried out during the austral summer season in the 49th Japanese Antarctic Research Expedition (JARE), 2007-2008. These studies were planned as one of the research projects named, "Studies on the changes of polar environments and ecosystems (P-3)" and the monitoring studies named "Monitoring for ecosystems (M-4)" during the 7th term of the Japanese Antarctic Research Expedition Plans. Field studies were done from 22 December 2007 to 13 February 2008, while our Ice Breaker Shirase stayed at/near off Syowa Station. To clarify the relationships among seasonal changes of environmental factors and biological responses, frequent field observations were performed at Naga Ike, one of the freshwater lakes in the Skarvsnes ice-free area. General limnological and biological samplings at the other lakes in the area (14 lakes near the Kizahasi Beach field base camp) were also done during the term. Observations and samplings distant from the base camp, four lakes in eastern Skarvsnes, a lake in Skallen, and three lakes in Langhovde, were also done using a helicopter for transportation. From Namazu Ike (temporary name) in eastern Skarvsnes, submersible video cameras were retrieved and so-called `algal crest\u27, benthic moss-algal assemblages, were sampled by scuba diving. Benthic copepods were sampled quantitatively from Nurume Ike in Langhovde. From Hyoga Ike (temporary name), a snow-dammed glacial lake which lost its water by recent breakage (during the JARE-46 wintering period), thin bio-film samples were collected from the present lake shore formerly part of the lake bed

Evolutionary histories of breast cancer and related clones

乳がん発生の進化の歴史を解明 --ゲノム解析による発がんメカニズムの探索--. 京都大学プレスリリース. 2023-07-28.Tracking the ol' mutation trail: Unraveling the long history of breast cancer formation. 京都大学プレスリリース. 2023-08-31.Recent studies have documented frequent evolution of clones carrying common cancer mutations in apparently normal tissues, which are implicated in cancer development1, 2, 3. However, our knowledge is still missing with regard to what additional driver events take place in what order, before one or more of these clones in normal tissues ultimately evolve to cancer. Here, using phylogenetic analyses of multiple microdissected samples from both cancer and non-cancer lesions, we show unique evolutionary histories of breast cancers harbouring der(1;16), a common driver alteration found in roughly 20% of breast cancers. The approximate timing of early evolutionary events was estimated from the mutation rate measured in normal epithelial cells. In der(1;16)(+) cancers, the derivative chromosome was acquired from early puberty to late adolescence, followed by the emergence of a common ancestor by the patient’s early 30s, from which both cancer and non-cancer clones evolved. Replacing the pre-existing mammary epithelium in the following years, these clones occupied a large area within the premenopausal breast tissues by the time of cancer diagnosis. Evolution of multiple independent cancer founders from the non-cancer ancestors was common, contributing to intratumour heterogeneity. The number of driver events did not correlate with histology, suggesting the role of local microenvironments and/or epigenetic driver events. A similar evolutionary pattern was also observed in another case evolving from an AKT1-mutated founder. Taken together, our findings provide new insight into how breast cancer evolves

Muscle Weakness and Fibrosis Due to Cell Autonomous and Non-cell Autonomous Events in Collagen VI Deficient Congenital Muscular Dystrophy

Congenital muscular dystrophies with collagen VI deficiency are inherited muscle disorders with a broad spectrum of clinical presentation and are caused by mutations in one of COL6A1–3 genes. Muscle pathology is characterized by fiber size variation and increased interstitial fibrosis and adipogenesis. In this study, we define critical events that contribute to muscle weakness and fibrosis in a mouse model with collagen VI deficiency. The Col6a1GT/GT mice develop non-progressive weakness from younger age, accompanied by stunted muscle growth due to reduced IGF-1 signaling activity. In addition, the Col6a1GT/GT mice have high numbers of interstitial skeletal muscle mesenchymal progenitor cells, which dramatically increase with repeated myofiber necrosis/regeneration. Our results suggest that impaired neonatal muscle growth and the activation of the mesenchymal cells in skeletal muscles contribute to the pathology of collagen VI deficient muscular dystrophy, and more importantly, provide the insights on the therapeutic strategies for collagen VI deficiency

Allele-specific Gene Silencing of Mutant mRNA Restores Cellular Function in Ullrich Congenital Muscular Dystrophy Fibroblasts

Ullrich congenital muscular dystrophy (UCMD) is an inherited muscle disorder characterized clinically by muscle weakness, distal joint hyperlaxity, and proximal joint contractures. Sporadic and recessive mutations in the three collagen VI genes, COL6A1, COL6A2, and COL6A3, are reported to be causative. In the sporadic forms, a heterozygous point mutation causing glycine substitution in the triple helical domain has been identified in higher rate. In this study, we examined the efficacy of siRNAs, which target point mutation site, on specific knockdown toward transcripts from mutant allele and evaluated consequent cellular phenotype of UCMD fibroblasts. We evaluated the effect of siRNAs targeted to silence-specific COL6A1 alleles in UCMD fibroblasts, where simultaneous expression of both wild-type and mutant collagen VI resulted in defective collagen localization. Addition of mutant-specific siRNAs allowed normal extracellular localization of collagen VI surrounding fibroblasts, suggesting selective inhibition of mutant collagen VI. Targeting the single-nucleotide COL6A1 c.850G>A (p.G284R) mutation responsible a sporadic autosomal dominant form of UCMD can potently and selectively block expression of mutant collagen VI. These results suggest that allele-specific knockdown of the mutant mRNA can potentially be considered as a therapeutic procedure in UCMD due to COL6A1 point mutations

Report on limnological, biological and ecological observations of lakes on the Soya Coast, East Antarctica

Observations on the limnological properties, samplings of waters and bottom assemblages for biological and ecological studies, and some field experimental studies at several lakes in Soya Coast ice-free areas, were carried out during the austral summer season in the 49th Japanese Antarctic Research Expedition (JARE), 2007-2008. These studies were planned as one of the research projects named, "Studies on the changes of polar environments and ecosystems (P-3)" and the monitoring studies named "Monitoring for ecosystems (M-4)" during the 7th term of the Japanese Antarctic Research Expedition Plans. Field studies were done from 22 December 2007 to 13 February 2008, while our Ice Breaker Shirase stayed at/near off Syowa Station. To clarify the relationships among seasonal changes of environmental factors and biological responses, frequent field observations were performed at Naga Ike, one of the freshwater lakes in the Skarvsnes ice-free area. General limnological and biological samplings at the other lakes in the area (14 lakes near the Kizahasi Beach field base camp) were also done during the term. Observations and samplings distant from the base camp, four lakes in eastern Skarvsnes, a lake in Skallen, and three lakes in Langhovde, were also done using a helicopter for transportation. From Namazu Ike (temporary name) in eastern Skarvsnes, submersible video cameras were retrieved and so-called `algal crest', benthic moss-algal assemblages, were sampled by scuba diving. Benthic copepods were sampled quantitatively from Nurume Ike in Langhovde. From Hyoga Ike (temporary name), a snow-dammed glacial lake which lost its water by recent breakage (during the JARE-46 wintering period), thin bio-film samples were collected from the present lake shore formerly part of the lake bed

Branchpoints as potential targets of exon-skipping therapies for genetic disorders

Fukutin (FKTN) c.647+2084G>T creates a pseudo-exon with a premature stop codon, which causes Fukuyama congenital muscular dystrophy (FCMD). We aimed to ameliorate aberrant splicing of FKTN caused by this variant. We screened compounds focusing on splicing regulation using the c.647+2084G>T splicing reporter and discovered that the branchpoint, which is essential for splicing reactions, could be a potential therapeutic target. To confirm the effectiveness of branchpoints as targets for exon skipping, we designed branchpoint-targeted antisense oligonucleotides (BP-AONs). This restored normal FKTN mRNA and protein production in FCMD patient myotubes. We identified a functional BP by detecting splicing intermediates and creating BP mutations in the FKTN reporter gene; this BP was non-redundant and sufficiently blocked by BP-AONs. Next, a BP-AON was designed for a different FCMD-causing variant, which induces pathogenic exon trapping by a common SINE-VNTR-Alu-type retrotransposon. Notably, this BP-AON also restored normal FKTN mRNA and protein production in FCMD patient myotubes. Our findings suggest that BPs could be potential targets in exon-skipping therapeutic strategies for genetic disorders