Volume 5 - 2018 | https://doi.org/10.3389/fmolb.2018.00044 This article is part of the Research TopicRNA Diseases in Humans – From Fundamental Research to Therapeutic ApplicationsView all 12 articles Fused in sarcoma (FUS) is an RNA binding protein that regulates RNA metabolism including alternative splicing FUS is genetically and pathologically involved in frontotemporal lobar degeneration (FTLD)/amyotrophic lateral sclerosis (ALS) Multiple lines of evidence across diverse models suggest that functional loss of FUS can lead to neuronal dysfunction and/or neuronal cell death Loss of FUS in the nucleus can impair alternative splicing and/or transcription whereas dysfunction of FUS in the cytoplasm especially in the dendritic spines of neurons Alternative splicing of the MAPT gene at exon 10 which generates 4-repeat Tau (4R-Tau) and 3-repeat Tau (3R-Tau) is one of the most impactful targets regulated by FUS loss of FUS function can affect dendritic spine maturations by destabilizing mRNAs such as Glutamate receptor 1 (GluA1) and Synaptic Ras GTPase-activating protein 1 (SynGAP1) FUS is involved in axonal transport and morphological maintenance of neurons These findings indicate that a biological link between loss of FUS function and phenotypic expression might lead to the sequential cascade culminating in FTLD to facilitate development of early disease markers and/or therapeutic targets of FTLD/ALS it is critical that the functions of FUS and its downstream pathways are unraveled This review provides an overview of recent findings that reveal the effects of functional loss of FUS on the pathogenesis of FTLD/ALS loss of FUS in the nucleus leads to imbalanced Tau isoforms due to insufficient skipping of exon 10 in the MAPT gene loss of FUS in the cytoplasm causes decreased stability in GluA1 and SynGAPα2 mRNA resulting in aberrant maturation of dendritic spines we summarize the roles of FUS in neurite maintenance and axonal transport and provide a briefly overview of the FUS liquid-phase-transition which may alter its various physiological functions and contribute to the development of toxic cellular effects under pathological conditions the functional properties of FUS may influence multiple cellular processes of neurons and/or glial cells whose dysfunction could be the most plausible explanation for neuronal toxicity mediated by loss of FUS the clinical data and FUS-silenced mice model findings support the hypothesis that FUS dysfunction results in early cognitive impairments These findings suggest that aberrant interactions between FUS and its spliceosome binding partners in the nucleus of neurons might lead to neuronal dysfunction and subsequent neurodegeneration future pathological studies examining the FUS/SFPQ nuclear interaction in both FTLD/ALS and tauopathies are necessary Proposed pathway underlying neurodegeneration following a qualitative loss of function of FUS in alternative splicing Under normal neuronal physiological conditions FUS and SFPQ interact in the nucleus to regulate alternative splicing of MAPT by skipping exon 10 When this functional machinery is impaired such as occurs following FUS or SFPQ depletion or qualitative loss due to disease-associated mutations or other unknown aberrant modifications the splicing ratio of MAPT exon 10+/exon 10- is increased which in turn results in an increased 4R-Tau/3R-Tau ratio The quantitative or qualitative loss of FUS or SFPQ causes various phenotypes Normalization of imbalanced Tau isoforms by co-injection with shRNA against 4R-Tau can successfully rescue these phenotypes our findings suggest that a pathophysiological link between FUS/SFPQ and the regulation of 4R-Tau/3R-Tau isoforms is involved in the pathogenesis of FTLD and 4R-tauopathy FUS also binds to the U-rich small nuclear ribonucleoproteins (U snRNPs) and SMN complexes in the spliceosome with disease-associated mutations in FUS affecting this alternative splicing machinery these findings indicate that both quantitative and qualitative losses of FUS function are likely involved in the pathogenesis of FTLD/ALS and should provide clues for therapeutics that clarify the functional properties of FUS The alteration of gene expression and/or alternative splicing of these genes may have a large impact on neuronal function which contributes to the neurodegeneration observed in FTLD/ALS We speculate that disruptions to FUS functionality could result in a partial effect rather than fatal damage by altering isoforms or expression levels of these genes it is possible that neurodegeneration only results after the transcriptional disruption triggered by loss of FUS functionality reaches a critical threshold such that the expression of individual genes and alternative splicing events are not critical by themselves Additional targets of FUS-mediated exon skipping could likewise contribute to FTLD/ALS pathogenesis. Among these genes is FUS itself in which FUS-mediated splicing at exon 7 contributes to autoregulation of expression with the exon 7 skipped variant undergoing nonsense-mediated decay (NMD). The auto regulatory function is deficient in ALS-associated FUS mutants (Zhou et al., 2013) Other FUS-regulated genes, such as NTNG1 or BRAF, which could be important for neuronal cell survival, have been identified in multiple reports (Orozco and Edbauer, 2013) Further study is necessary to evaluate their significance in FTLD/ALS pathogenesis these findings suggest that the cytoplasmic function of FUS may be involved in regulating mRNA stability suggest that the loss of regulatory control of synaptic molecule mRNA stability in response to impaired FUS functionality causes synaptic dysfunction and could lead to post-synapse impairments in FTLD/ALS the morphological abnormalities in neurites might be one of the earliest biomarkers and could thus be used in therapeutic screens or as a diagnostic tool further investigation to clarify the common downstream pathomechanism is necessary These findings strongly suggest a biochemical link between RNA-binding proteins and other amyloid-formable proteins including Tau and its association with RNA processing in neurodegenerative diseases Since those findings were based on in vitro experiments further investigation is necessary to clarify whether/how liquid-liquid phase transitions are associated with biological function and whether transitions that occur in the cytoplasm of dendritic spines and/or the nucleus utilize the same or a different molecular process to determine the utility of FUS and its downstream pathways as early disease markers and/or therapeutic targets of FTLD/ALS it is crucial that their functional properties be more precisely clarified Functional loss of FUS in the nucleus and the cytoplasm can cause neuronal dysfunction and degeneration FUS regulates alternative splicing and transcription which is regulated by FUS in complex with SFPQ FUS also regulates transcription of a number of genes including Ntng1 cytoplasmic FUS stabilizes mRNAs involved in the dendritic spine the functional impairments caused by FUS deficiency can affect neuronal function and morphology and subsequently lead to aberrant behaviors and neurodegeneration FUS has also been implicated in the axon transport machinery which is impaired by disease-associated mutations in FUS The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest This work was supported by Mext Grant–in-aid project Scientific Research on Innovation Area (Brain Protein Aging and Dementia control) by Mext Grant-in-Aid for Scientific Research on Innovative Areas (Comprehensive Brain Science Network) This work was also supported by the Integrated Research on Neuropsychiatric Disorders and Integrated Research on Depression Dementia and Development Disorders projects carried out under the Strategic Research Program for Brain Sciences and Brain/MINDS of the Japan Agency for Medical Research and Development Liquid-liquid phase separation of the microtubule-binding repeats of the Alzheimer-related protein Tau Localization of fused in sarcoma (FUS) protein to the post-synaptic density in the brain Loss and gain of FUS function impair neuromuscular synaptic transmission in a genetic model of ALS Axonal transport defects are a common phenotype in Drosophila models of ALS Tau-mediated neurodegeneration in Alzheimer's disease and related disorders Juvenile ALS with basophilic inclusions is a FUS proteinopathy with FUS mutations Novel FUS deletion in a patient with juvenile amyotrophic lateral sclerosis a novel RNA/ssDNA-binding protein with homology to the pro-oncoproteins TLS/FUS and EWS is associated with both TFIID and RNA polymerase II Phase separation of C9orf72 dipeptide repeats perturbs stress granule dynamics Boillée Onset and progression in inherited ALS determined by motor neurons and microglia PINK1 and parkin are genetic modifiers for FUS-induced neurodegeneration Liquid-liquid phase separation in cellular signaling systems ALS mutations in TLS/FUS disrupt target gene expression TDP-43 and FUS RNA-binding proteins bind distinct sets of cytoplasmic messenger RNAs and differently regulate their post-transcriptional fate in motoneuron-like cells Fusion of CHOP to a novel RNA-binding protein in human myxoid liposarcoma FUS-immunoreactive inclusions are a common feature in sporadic and non-SOD1 familial amyotrophic lateral sclerosis Silencing of FUS in the common marmoset (Callithrix jacchus) brain via stereotaxic injection of an adeno-associated virus encoding shRNA CrossRef Full Text FUS affects circular RNA expression in murine embryonic stem cell-derived motor neurons Familial behavioral variant frontotemporal dementia associated with astrocyte-predominant tauopathy TLS facilitates transport of mRNA encoding an actin-stabilizing protein to dendritic spines FUS-regulated region- and cell-type-specific transcriptome is associated with cell selectivity in ALS/FTLD Dysregulated molecular pathways in amyotrophic lateral sclerosis-frontotemporal dementia spectrum disorder HDAC6 inhibition reverses axonal transport defects in motor neurons derived from FUS-ALS patients RNA-binding proteins with prion-like domains in health and disease Dendritic spine pathology in Neurodegenerative diseases Fus deficiency in mice results in defective B-lymphocyte development and activation high levels of chromosomal instability and perinatal death The ALS/FTLD-related RNA-binding proteins TDP-43 and FUS have common downstream RNA targets in cortical neurons Mutation-specific functional impairments in distinct tau isoforms of hereditary FTDP-17 Tau mislocalization to dendritic spines mediates synaptic dysfunction independently of neurodegeneration Extensive FUS-immunoreactive pathology in juvenile amyotrophic lateral sclerosis with basophilic inclusions Establishment of in vitro FUS-associated familial amyotrophic lateral sclerosis model using human induced pluripotent stem cells Altered tau isoform ratio caused by loss of FUS and SFPQ function leads to FTLD-like phenotypes Position-dependent FUS-RNA interactions regulate alternative splicing events and transcriptions FUS and TARDBP but not SOD1 interact in genetic models of amyotrophic lateral sclerosis Distinct and shared functions of ALS-associated proteins TDP-43 FUS and TAF15 revealed by multisystem analyses Intracellular localization and splicing regulation of FUS/TLS are variably affected by amyotrophic lateral sclerosis-linked mutations FUS/TLS deficiency causes behavioral and pathological abnormalities distinct from amyotrophic lateral sclerosis Male sterility and enhanced radiation sensitivity in TLS(−/−) mice Mutations in the FUS/TLS gene on chromosome 16 cause familial amyotrophic lateral sclerosis Lagier-Tourenne Lagier-Tourenne Divergent roles of ALS-linked proteins FUS/TLS and TDP-43 intersect in processing long pre-mRNAs Pathological heterogeneity in amyotrophic lateral sclerosis with FUS mutations: two distinct patterns correlating with disease severity and mutation Position-specific binding of FUS to nascent RNA regulates mRNA length Phosphorylation of the FUS low-complexity domain disrupts phase separation ALS/FTD mutation-induced phase transition of FUS liquid droplets and reversible hydrogels into irreversible hydrogels impairs RNP granule function Structure of FUS protein fibrils and its relevance to self-assembly and phase separation of low-complexity domains FUS regulates genes coding for RNA-binding proteins in neurons by binding to their highly conserved introns A new subtype of frontotemporal lobar degeneration with FUS pathology FUS-mediated alternative splicing in the nervous system: consequences for ALS and FTLD Loss of fused in sarcoma (FUS) promotes pathological Tau splicing Differential regulation of microtubule dynamics by three- and four-repeat tau: implications for the onset of neurodegenerative disease A liquid-to-solid phase transition of the ALS protein FUS accelerated by disease mutation ALS-associated mutation FUS-R521C causes DNA damage and RNA splicing defects State of play in amyotrophic lateral sclerosis genetics Lower motor neuron involvement in TAR DNA-binding protein of 43 kDa-related frontotemporal lobar degeneration and amyotrophic lateral sclerosis The changing scene of amyotrophic lateral sclerosis Widespread binding of FUS along nascent RNA regulates alternative splicing in the brain Knockdown of the Drosophila fused in sarcoma (FUS) homologue causes deficient locomotive behavior and shortening of motoneuron terminal branches Scekic-Zahirovic Toxic gain of function from mutant FUS protein is crucial to trigger cell autonomous motor neuron loss Super-resolution microscopy reveals presynaptic localization of the ALS/FTD related protein FUS in hippocampal neurons FUS binds the CTD of RNA polymerase II and regulates its phosphorylation at Ser2 genetic and pathological heterogeneity of frontotemporal dementia: a review Activity-dependent FUS dysregulation disrupts synaptic homeostasis The function of RNA-binding proteins at the synapse: implications for neurodegeneration ALS-associated mutant FUS induces selective motor neuron degeneration through toxic gain of function TDP-43 and FUS/TLS: sending a complex message about messenger RNA in amyotrophic lateral sclerosis ALS-causative mutations in FUS/TLS confer gain and loss of function by altered association with SMN and U1-snRNP The phenotypic variability of amyotrophic lateral sclerosis TLS/FUS (translocated in liposarcoma/fused in sarcoma) regulates target gene transcription via single-stranded DNA response elements Non-nuclear pool of splicing factor SFPQ regulates axonal transcripts required for normal motor development Spliceosome integrity is defective in the motor neuron diseases ALS and SMA FUS regulates AMPA receptor function and FTLD/ALS-associated behaviour via GluA1 mRNA stabilization Neurodegenerative disorder FTDP-17-related tau intron 10 +16C –> T mutation increases tau exon 10 splicing and causes tauopathy in transgenic mice cause familial amyotrophic lateral sclerosis type 6 TDP-43 loss-of-function causes neuronal loss due to defective steroid receptor-mediated gene program switching in Drosophila Van Langenhove The molecular basis of the frontotemporal lobar degeneration-amyotrophic lateral sclerosis spectrum The ALS-associated proteins FUS and TDP-43 function together to affect Drosophila locomotion and life span Induced ncRNAs allosterically modify RNA-binding proteins in cis to inhibit transcription Slowing of axonal transport is a very early event in the toxicity of ALS-linked SOD1 mutants to motor neurons The LC Domain of hnRNPA2 adopts similar conformations in hydrogel polymers Mutant SOD1 in cell types other than motor neurons and oligodendrocytes accelerates onset of disease in ALS mice Astrocytes as determinants of disease progression in inherited amyotrophic lateral sclerosis Sporadic juvenile amyotrophic lateral sclerosis caused by mutant FUS/TLS: possible association of mental retardation with this mutation Frameshift and novel mutations in FUS in familial amyotrophic lateral sclerosis and ALS/dementia The RNA-binding protein Fus directs translation of localized mRNAs in APC-RNP granules 3'UTR length-dependent control of SynGAP isoform α2 mRNA by FUS and ELAV-like proteins promotes dendritic spine maturation and cognitive function Cellular tau pathology and immunohistochemical study of tau isoforms in sporadic tauopathies ALS-associated FUS mutations result in compromised FUS alternative splicing and autoregulation Citation: Ishigaki S and Sobue G (2018) Importance of Functional Loss of FUS in FTLD/ALS Received: 28 February 2018; Accepted: 17 April 2018; Published: 03 May 2018 Copyright © 2018 Ishigaki and Sobue. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) distribution or reproduction in other forums is permitted provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited in accordance with accepted academic practice distribution or reproduction is permitted which does not comply with these terms *Correspondence: Shinsuke Ishigaki, aXNoaWdha2ktbnNAdW1pbi5uZXQ= Gen Sobue, c29idWVnQG1lZC5uYWdveWEtdS5hYy5qcA== Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish （左から）祖父江さん、志間さん、ドミニクさん。届いたばかりの書籍が入って“いた”段ボールに囲まれた（笑）祖父江さんのデザイン事務所「コズフィッシュ」にて 早稲田駅、飯田橋方面ホームにある広告。SJC学生スタッフの田邊さん（文化構想学部4年）が記念撮影 2024年春、東京メトロ東西線早稲田駅ホームに、『早稲田ウィークリー』の広告が掲示されました。デザインを担当したのは、夏目漱石『心』やさくらももこの著書をはじめ、小説、漫画など奇想天外かつ常識を覆す装丁を世に送り続ける、デザイン界の巨匠・祖父江慎さん。そのデザインは、拍子抜けするほどシンプルで、随所に“ヘン”なバランスが潜んでいます。 今回は、『早稲田ウィークリー』広告の違和感のナゾやデザインに込めた思いについて、祖父江さんにお聞きしようとスペシャル対談を企画。対談相手は、長年祖父江さんを敬愛し、自身がディレクターを務める展覧会でもデザインをお願いしたというドミニク・チェン教授（文学学術院）。さらに、祖父江さんと共にウィークリーの新しいロゴデザインに携わった、デザイナーの志間かれんさんにも加わっていただきました。 生粋の自由人であり、「わかりにくさ」や「うまくいかなさ」の中に面白さを見いだす達人たちの話は、『早稲田ウィークリー』の広告から、自由に生きるための秘訣(ひけつ)にまで及んで…。さあ、縦横無尽な対談の始まりです。 ドミニク：祖父江さんの存在を初めて知ったきっかけは、9歳くらいのときに読んだ吉田戦車さんの漫画『伝染るんです。』。子ども心に「何だこのハチャメチャなブックデザインは！」とドキドキしたことを覚えています。だから2020年に、21_21 DESIGN SIGHTで僕がディレクションした「トランスレーションズ展」で祖父江さんの事務所コズフィッシュにグラフィックデザインを引き受けていただけたときは、天にも昇る気持ちでした（笑）。そんな祖父江さんが今回、早稲田ウィークリーの駅広告をデザインしたと聞いて、うれしさと同時に驚きもありました。どうして引き受けてくださったのですか？ 祖父江：僕ね、やったことないことって、無性にやりたくなるんです。まず、大学が作る媒体のデザインが初めてだった。駅広告のデザインは何度かやったことがあるのだけど、それらは自分が関わった展覧会の広告など。『早稲田ウィークリー』の仕事は、そういう流れがあるわけでなく、突然早稲田駅のホームに掲示する広告だけを依頼されたんです。その唐突さが面白いなぁって思ったんです。 ドミニク：広告を見て最初に「おお、文章がメインなのか」と思いました。大学関係の媒体って、キャンパスや学生の写真を多く使うのに、そういうのが一切ないのが新鮮ですね。 祖父江：そう、まずは文章を読んでほしいと思って。そのために、写真を使用せず、WASEDA BEAR（以下、ワセダベア）のイラストとエンブレムのみ、文字数も最小限に、本当に伝えたい内容だけを載せています。 ドミニク：よーく見ると、真ん中の「早稲田ウィークリー」の文字だけフォントが違う？  一見普通の明朝体のようで、文字の周りがにじんでいるように見えます。これは意図的なんですか？ 祖父江：もっとよく見てみて。実は文字ごとに違うんですよ。ひらがなやカタカナはにじませているけど、漢字はそうじゃない。あえて統一しないことで、「なんで？」とツッコミどころが生まれる。するとね、そこから会話が生まれるんです。 祖父江 慎（そぶえ・しん）愛知県生まれ。多摩美術大学グラフィックデザイン科を中退後、グラフィックデザイナー、アート・ディレクターとして活躍しながら、現在、コズフィッシュ代表。日本のブックデザインの最前線で幅広いジャンルを手掛けている。スヌーピーミュージアム東京、ミッフィー展などの展覧会グラフィック、グッズデザインにも力を発揮する。過去の仕事をまとめた『祖父江 慎＋cozfish』（PIE BOOKS）。「いつか祖父江さんと仕事をするのが夢」と編集者を志す学生も多数 ドミニク：おお、ほんとだ！ 祖父江さんからデザインの授業を受けているようでうれしいです（笑）。あと、デザインとは関係ないのですが、広告の文字を見て、早稲田ウィークリーが創刊59年だということにも驚きました。 祖父江：中途半端な数字で面白いですよね。他の広告ではなかなかお目にかかりません（笑）。初の広告を出すなら、あと1年待って「60年」にすればいいのにって。 創刊当時の『早稲田ウィークリー』は週1回刊行される紙の新聞だったそう。それが8年前からWebマガジンになって、授業期間中は平日毎日更新している。なら、「デイリー」じゃんとか、創刊59年「Webマガジン」ってインターネットがない時代じゃんとか。大学の媒体なのに、ツッコミどころ満載なんです。そういうのを見つけると、誰かにしゃべりたくなるでしょ。 ドミニク：とはいえ、広告内でツッコミどころについて細かく説明するわけでもない。どう捉えるかは、見る人にお任せしていますね。 祖父江：クレームが付かないよう、もしもの場合に備えた説明や注釈ばかり付け加えるのは好きじゃないんです。そうすると情報過多の、押し付けがましい、嫌われ者の広告になってしまいます。 それよりも、伝えたいと思ったことが、人それぞれの捉え方をされて伝わる方が面白いじゃない？ うまく伝わったり伝わらなかったりするのは、誰かとやりとりできているということだから、その「うまくいかなさ」を楽しんだ方がいいんです！ ドミニク：大学で教えていると同じことを感じます。僕は教員になって7年目になるのですが、「教授」や「教員」っていう字面が苦手で。決まってる正解を教えないといけない、伝えないといけないという気持ちが生まれて苦しくなります。反対に、面白いなと感じるのは、僕が想定したことが良い感じに誤読されたときなんですよ。こちらの想像の斜め上を行かれるときほどうれしい（笑）。 祖父江：デザイナーも一緒ですよ。生きる上でも、うまくいかなさを面白がる力が大事なんじゃないかな。 ドミニク：もう一つ、右下にある「早稲田ウィークリー」のロゴが面白いですね。「W」の形が不揃いで、全然別の方向を向いているのが早稲田っぽくていい（笑）。 新しいロゴとウィークリー仕様の早稲田ベア。スマホの画面は、ウィークリーのテーマカラーのライトグリーンに 祖父江：そうそう、ついでに新しいロゴも作って、ワセダベアもウィークリー仕様に微調整しました。ロゴはデザイナーの志間さんがいろいろと試して作ってくれて、「W」はこの向きが一番良かった。ぼくは早稲田出身じゃないけど、早稲田って自由な大学じゃないですか。活動的な学生たちが、みんなそれぞれ好きなことをやっている。それが二つの「W」のてんでバラバラな感じに表れていると思います。 志間：私も違う大学の出身ですが、大学へ進学した同級生たちは、明るくキャンパスライフを満喫している印象を持っていました。だから、ロゴのポイントとなる「W」の文字も、のびのび自由に動かしてみたんです。一つめのWと二つめのWの幅も意図的に変えています。 ドミニク：なるほど、一つだけの正解に向かうんじゃなくて、異質な人が同じ場所に集まってる感じがして、すごい好きです。ワセダベアはどんな風に変えたのですか？ 祖父江：もともとは、がっしりした体格の、いかにも男っぽいキャラクターです。つえを持っていることもあるし、まるで先生みたい。ウィークリーのワセダベアはもっと親しみやすい方がいいと思ったんです。 志間：そこで、大きく印象が変わらないように気を付けながら、クッキーのようなおいしそうな色使いにして、スマホを持たせて、学生らしさを出しました。眉毛はちょっと気弱な角度にしています。目も横を向かせて、口元も困っているみたいにしました。 祖父江：いわば昭和っぽいベアから令和のベアへ変身です。でもその方が、距離は縮まりますよね。 ドミニク：どこか困ってるような、弱さが出ちゃってるベアの方が愛着や親近感が持てますね。それと、左のエンブレム。主張する気の無さそうな控えめな大きさで、宙に浮いているような、不思議な位置にあって、どうしても気になってしまいます。 祖父江：エンブレムについては、実は早稲田ウィークリー編集室から、「不思議すぎる位置だから、真ん中とか左下のわかりやすい場所に移動した方がいいのでは」と何度も相談を受けました。でもね、左下とか真ん中って一番目立ちそうでいて、一番人の目に留まらない場所なんです。みんな、当たり前のように真ん中に置いてしまうことの危険性をもっと考えた方がいい（笑）。 かといってテキトーに配置したわけではなく、縦の位置は最後の2行の行間に配置してあって、配置に論理はあるんです。でも一般的な論理ではない。そうすることで、見た人に考える喜びが生まれるんです。ドミニクさんみたいに、これはどうしてここにあるんだろう、どうしてこんなに小さいのだろうって。考える喜びが生まれて、見れば見るほど味わいが出てくる、俳句みたいな広告なんです。ぜひ、皆さんも毎日の通学で眺めてみてください。 この大学には、自由でのびのびとした人がたくさん集まっている。教えようとしても、みんな自分のやり方で勝手に動いていってしまう。今回の『早稲田ウィークリー』の広告には、そんな早稲田大学の素敵さを出すことができたかなと思います。 ドミニク：僕は子どもの頃に祖父江さんのブックデザインに出合ったことで、「自由ってこういうことか！」と教えてもらったと思ってて、祖父江さんのように自由でのびのびと生きるための秘訣(ひけつ)についてもお聞きしたいです。早稲田は多様な学生が多い校風の中で、無理に個性を出そうとして、かえって苦しくなってしまう学生も多いような気がします。 祖父江：時々インタビューなどで「どうしてこのような個性的なデザインにしたのですか」と聞かれることがあります。でも、自分を出したいとか、無理やりにでもこれを伝えたいとか、そういうことは考えてないんです。 大事にしているのは、「うっとり」すること。それは、われを忘れて何かに浸ってしまう時間のこと。そこから思いも寄らなかった面白いものが出てきます。だから、僕がやっているのは、自分を出すのではなく、自分を忘れていくようなデザインなんです。 ドミニク：「自分を忘れてしまう」って重要ですよね。でも、そのためには注意深い観察が必要な気もします。僕のゼミでは今、みんなで日常生活の定点観測をしています。ふとした景色に心を動かされたとか、机の模様が気に入ったとか、そういうすぐに記憶から消えてしまいそうなものを写真に撮ったり文章に記録したりして、自分なりの感覚や物の見方を見つけてみよう、と。それは言い換えれば、自分が自分であることの証拠集めをしているということなんですよね。そうすると、自分や周りの世界の輪郭がおのずから浮かび上がってくる感覚があります。 ドミニク・チェン 早稲田大学文学学術院教授。博士（学際情報学）。文化構想学部の表象・メディア論系で発酵メディア研究ゼミを主催し、しゃべるぬか床Nukabotの開発や「発酵」をコンセプトとしたデザインワークショップを開催しながら、テクノロジーと人間、そして自然存在の望ましい関係性を研究している。2020年に、21_21 DESIGN SIGHT『トランスレーションズ展―「わかりあえなさ」をわかりあおう』の展示ディレクターを務め、コズフィッシュにグラフィックデザインをお願いした。著書に『未来をつくる言葉 わかりあえなさをつなぐために』（新潮社）など多数 祖父江：そのときは必要なさそうなものが、後から見れば実はものすごく偉大で尊いものなんですよね。それに気付くと、自分や世界がより分かるようになる。でもその発見は自分ではコントロールできない。だからこそうれしさがあります。 ドミニク：なるほど。「完全にコントロールできないけど自分が関わりながら変化している」過程のことを、僕は「発酵」と呼んでいます。ゼミの名前も「発酵メディア研究ゼミ」にしてます（笑）。というのも、自分自身についての発見は、ぬか床で漬物を作るのに似ているんです。漬物の味は、入れる野菜の種類でも変わるし、ぬか床の置く場所でも変化します。手入れしないとカビが生えたり、きちんとケアすると良い香りが生まれたりするけれども、かといって味を完全にコントロールできるわけでもない。 自分の個性を見つける過程も、野菜をぬか床に入れて、どう発酵していくか、どんな味になるかと眺めている感じと一緒だと思います。 祖父江：そう、眺める感じだよね。僕の言葉では、「われを忘れて、うっとりしてしまう状態」。でも、どれだけ自分を忘れても、残り続けるクセみたいなものがある。忘れたいけど残ってしまう、失くしたいけど残ってしまう。でも、なんだか許せちゃうものが自分の個性になるんです。 祖父江さんが「われを忘れる」ものの一つが「文字で表現されているもの」。『坊っちゃん』から始まり、『ピノッキオ』『南総里見八犬伝』は、同じタイトルのものを収集して比べている。写真は、世界中の『ピノッキオ』を集めた書棚。文字や絵、ビジュアル表現の変遷だけでなく、本ごとのストーリーの変わり方にも注目して読んでいるとか ドミニク：ああ、「許せちゃう」っていい言葉ですね。はっきりと「良い／悪い」で区切ったものだけだと、結局は予定調和になってしまう。けど、うまくいかないことを許せると、心に厚みがでるというか、容量が増えるイメージがしますね。 祖父江：良くないなと思っていたものが、いつのまにか自分の個性だと感じるようになる。そんな状況を楽しめることが、生きるということの喜びでもあるんじゃないのかな。20歳くらいの人たちがわれを忘れて何かに熱中するからこそ生まれてくる面白さを、学生の皆さんには大事にしていってほしいなと思います。 早稲田大学オフィシャルサイト（https://www.waseda.jp/inst/weekly/）は、以下のWebブラウザでご覧いただくことを推奨いたします。 推奨環境以外でのご利用や、推奨環境であっても設定によっては、ご利用できない場合や正しく表示されない場合がございます。より快適にご利用いただくため、お使いのブラウザを最新版に更新してご覧ください。 このままご覧いただく方は、「このまま進む」ボタンをクリックし、次ページに進んでください。 このまま進む  Google ChromeWindowsバージョン38 以上Macintoshバージョン38 以上Webサイト Fire FoxWindowsバージョン33 以上Macintoshバージョン33 以上Webサイト SafariWindowsバージョン38 以上Macintoshバージョン38 以上Webサイト Internet ExplorlerWindowsバージョン10 以上Webサイト Volume 13 - 2019 | https://doi.org/10.3389/fncir.2019.00074 Human brain imaging studies have revealed several regions that are activated in patients with chronic pain functional changes due to chronic pain have not been fully elucidated as brain imaging techniques such as functional magnetic resonance imaging and positron emission tomography (PET) require the use of anesthesia to suppress movement conclusions derived from existing imaging studies in rodents may not accurately reflect brain activity under awake conditions we used quantitative activation-induced manganese-enhanced magnetic resonance imaging to directly capture the previous brain activity of awake mice We also observed and quantified the brain activity of the spared nerve injury (SNI) neuropathic pain model during awake conditions SNI-operated mice exhibited a robust decrease of mechanical nociceptive threshold 14 days after nerve injury Imaging on SNI-operated mice revealed increased neural activity in the limbic system and secondary somatosensory We present the first study demonstrating a direct measurement of awake neural activity in a neuropathic pain mouse model direct measurement of neural activity in awake resting-state rodents has not been successful The present study investigates the steady-state brain activity of neuropathic pain in awake mice. AIM-MRI with quantitative T1 measurement (qAIM-MRI), quantifying T1 values (Tambalo et al., 2009; Kikuta et al., 2015), was used on a spared nerve injury (SNI) model of neuropathic pain (Yamamoto et al., 2016) We found that steady-state brain activity in an awake neuropathic pain mouse model showed increased activity of the prefrontal-limbic-basal ganglia circuit this study is first to directly measure neural activity in animals with neuropathic pain under awake conditions All animal studies were approved by the Animal Care Committee of the Graduate School of Pharmaceutical Sciences Nagoya City University and by the Tohoku University Committee for Animal Experiments Experiments were conducted in accordance to the guidelines of the National Institute of Health and the Japanese Pharmacological Society We used male C57BL/6J mice (6 weeks old; CLEA Japan All mice were housed in a room maintained at 23 ± 2°C with an alternating 12-h light-dark cycle and had ad libitum access to food and water The surgical procedure for producing the SNI model was originally described by Decosterd and Woolf (2000) Animals were anesthetized with isoflurane (4% for induction An incision was made in the skin on the lateral surface of the left thigh followed by a section through the biceps femoris muscle to expose the sciatic nerves The common peroneal and tibial nerves were then tightly ligated with 8–0 silk suture and 1 mm of the distal nerve stump was cut Sham-operated controls were subjected to exposure of the sciatic nerve and its branches without any lesions All measurements were carried out in a 9.4 T MRI scanner (AV400WB Bruker) equipped with a 45 G/cm gradient and a 38 mm 1H volume coil (Bruker) The total time required to obtain all magnetic resonance images was approximately 45 min MRI analysis methods were also described previously (Kikuta et al., 2015) Brief methods are as follows: after spatial filtering parametric T1 maps were calculated pixel-by-pixel by fitting with the following equation using Para Vision 5.1 software (Bruker BioSpin) where SI is signal intensity in each pixel we could then identify brain regions by querying structures from the brain atlas An unpaired Student’s t-test was used to determine which T1 voxels decreased or increased in the SNI group compared with the sham group using SPM12 A parametric map of voxels with statistically significant changes in T1 was created and overlaid on the T2-weighted template image ROIs were set in the area where significant changes in T1 values were observed in SPM analysis The ROI size was 3 × 3 × 3 voxels Average T1 values in ROIs were calculated for statistical analysis Statistical analysis was performed using R For the statistical parametric mapping (SPM) analysis statistical significance (p < 0.025) was assessed by the unpaired Student’s t-test using SPM12 software To ascertain the variability of T1 values within each group and if a significant difference (p < 0.05) was detected we confirmed the significance of the mean T1 values by bootstrapping within a 95% confidence interval after 1,000 randomizations All data are presented as mean ± standard error of mean Fourteen days after SNI surgery, mice exhibited a prominent decrease of mechanical nociceptive threshold in the injured paw (Figure 1). In contrast, sham-operated mice did not exhibit the reduction of mechanical nociceptive threshold (Figure 1) Mechanical nociceptive threshold of the hind paw in spared nerve injury (SNI) and sham-operated mice 14 days after nerve ligation The SNI group exhibited decreased mechanical nociceptive threshold compared with the sham-operated group Each column represents the mean ± SEM from 5 (sham group) and 8 (SNI group) we conducted Mann-Whitney U-test and bootstrap analysis of the mean T1 value in each ROI Significant shortening of T1 values were detected in NAc (Sham; 2.35 ± 0.04 The active regions within the limbic structure in SNI mice compared with sham mice analyzed by AIM-MRI Regions with significant shortening of T1 in SNI mice are indicated by the pseudo-colored regions over the T2-enhanced brain image template in coronal planes (n = 6 for sham The active regions within the limbic structure are defined in text: nucleus accumbens (NAc) and ventral posterolateral nucleus of thalamus (VPL) The distance from bregma in mm is shown at the bottom The active regions in the cortex in SNI mice compared with sham mice analyzed by AIM-MRI The active regions in the cortex are defined in text: piriform cortex (Pir) and T1 value of the active region in SNI compared with sham mice (n = 6) analyzed by AIM-MRI The present results indicate that the neuropathic pain mouse model experienced an increase of resting-state brain activity and that qAIM-MRI can successfully distinguish neural activity in both the neuropathic pain mouse model and normal control mice the animals could still be anesthetized during MRI for visualization of brain activity The qAIM-MRI method used in this study enables quantitative neuronal activity mapping over the entire brain and can help characterize changes to brain activity in neuropathic pain There are several reasons behind excluding measurement from the PAG and cerebellum in this study since our major focus in this study was to explore the function of the frontal part of cerebral cortex in pain processing the hind brain regions such as PAG and cerebellum were not evaluated Further studies are required to reveal the activation of the PAG and cerebellum under chronic pain we also observed increased activation of the prefrontal-limbic-basal ganglia network during resting-state brain activity in SNI mice using qAIM-MRI Since the qAIM-MRI method directly determines the neuronal activity in freely moving animals our present results provide comprehensive evidence that the regions in the nervous system related to emotion might be activated under neuropathic pain and insular cortex were also activated 8 days after SNI surgery we observed the activation of several limbic structures in SNI mice 14 days after surgery Since the duration of chronic pain might differentially affect neural plasticity in each brain region the functional changes across brain regions might be widespread upon prolongation of chronic pain The advantage of qAIM-MRI lies in its ability to measure the absolute T1 value, as opposed to several studies using MEMRI that measures the signal intensities obtained from T1-weighted images (Chao et al., 2018) T1-weighted images are quantified by signal intensities that provide relative values and therefore can be unreliable for comparisons between animals can be used to quantify the Mn2+ concentration our present results reflect absolute neural activation under chronic pain intravenous administration of MnCl2 at a dose of 75 mg/kg seems very high to evaluate the brain activity by MEMRI Mn2+ accumulation in astrocytes may correlate with the activity of adjacent neurons changes to neural activity in areas that process pain cognition and emotion contribute to the chronification of neuropathic pain Nagoya City University and the Tohoku University Committee for Animal Experiments and MOh performed the experiments and data analysis MOs and MOh calculated the statistical significance and wrote the manuscript This study was supported by the KAKENHI Grant Numbers 16H05460 (MOh) and 17H05543 (MOs) from the Ministry of Education and Brain/MINDS [Mapping by Integrated Neurotechnologies for Disease Studies (JP19dm0207001)] We thank the Institute for Animal Experimentation and Biomedical Research Core Graduate School of Medicine Tohoku University for the permitting the use of its facilities and their technical assistance Manganese-enhanced magnetic resonance imaging (MEMRI) of brain activity and applications to early detection of brain ischemia Human brain mechanisms of pain perception and regulation in health and disease Neuropathic pain symptoms relative to overall pain rating The use of the transition elements manganese cobalt and nickel as synaptic blocking agents on isolated CrossRef Full Text | Google Scholar Chronic pain and the emotional brain: specific brain activity associated with spontaneous fluctuations of intensity of chronic back pain Corticostriatal functional connectivity predicts 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mechanical hypersensitivity Current status of functional MRI on small animals: application to physiology Clinical and pre-clinical pain assessment: are we measuring the same thing Ultrasound vocalisation by rodents does not correlate with behavioural measures of persistent pain N- and L-type calcium channels blocker cilnidipine ameliorates neuropathic pain In vivo auditory brain mapping in mice with Mn-enhanced MRI Osanai M and Ohsawa M (2019) Visualization of Brain Activity in a Neuropathic Pain Model Using Quantitative Activity-Dependent Manganese Magnetic Resonance Imaging Copyright © 2019 Inami, Tanihira, Kikuta, Ogasawara, Sobue, Kume, Osanai and Ohsawa. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited *Correspondence: Masahiro Ohsawa, b2hzYXdhQHBoYXIubmFnb3lhLWN1LmFjLmpw †These authors have contributed equally to this work Japanese cross-country skiers Ryo Hirose and Rin Sobue won the mixed team sprint classic today securing the first gold medal of the Lake Placid 2023 FISU World University Games American athletes Finn Sweet and Renae Anderson opened Team USA’s medal count with an impressive silver at the end of an intense battle with bronze medallist Norway whose words really set the tone of the intense joy from the Japanese team The skiers from Waseda University controlled the sprint final from beginning to end never letting a chance to their competitors Hirose and Sobue met the expectations put on their shoulders finishing with a near 10-second lead over the USA After finishing fourth in their semi-final the Japanese favorites stood up during the final using the experience they gained from their participation at the 2022 Beijing Olympic Games The nippon team singularly took advantage of the intense weather conditions and the falling snow during the final which made the tracks slower and the race even more tense than during the semi-finals “It’s very similar to what we know in Japan,” Sobue explained with a large smile on her face after the race But the surprise of this event came from Team USA’s second pair which also managed to step up in these tough conditions as local fans were incredibly excited to see the American duo get the silver medal at the first event of these home Games “Our mindsets were in the right place for toughness,” said Anderson after an impressive finish that allowed her team to edge the Norwegian squad Cheering crowds also witnessed a glimpse of the fast-changing weather conditions in Lake Placid from rain in the early morning to intense snow for the final This has been acknowledged as a critical topic by the Organizing Committee as the Save Winter FISU World Conference takes place over the next three days while athletes are competing The Japanese team hopes these conditions will continue to help them as they were not afraid to express their ambitions after the mascot ceremony “We want to win five medals in all five races,” they said jointly © International University Sports Federation The dates displayed for an article provide information on when various publication milestones were reached at the journal that has published the article activities on preceding journals at which the article was previously under consideration are not shown (for instance submission All content on this site: Copyright © 2025 Elsevier B.V. Researchers find that kidney dysfunction can be used as a criterion to stratify patients requiring implantable defibrillators This study by Fujita Health University researchers revealed that kidney function considered in terms of estimated glomerular filtration rate (eGFR) can be used as a predictor for SCD in patients with congestive heart failure Image courtesy of Yoshihiro Sobue from Fujita Health University 2024 — Patients with congestive heart failure (CHF) having a compromised blood supply are at greater risk of sudden cardiac death (SCD) With an estimated incidence as high as 22% among these patients current clinical guidelines recommend using implantable cardioverter defibrillators (ICDs) to help mitigate the risk of SCD Although the application of ICDs in patients with CHF is backed by substantial evidence these studies did not include people with chronic kidney disease (CKD) it is unclear whether the use of ICDs is justified in patients with CKD especially with mild and moderate presentations given the inherent risk involved in installing these devices and Eiichi Watanabe from Fujita Health University Bantane Hospital Elaborating on the rationale behind this examination “The objective of our study was to prospectively investigate the risk factors for SCD in a cohort of over 1,500 patients hospitalized with CHF classified based on the left ventricular ejection fraction (LVEF) and the New York Heart Association (NYHA) functional class as well as the potential role of kidney function as a determinant of SCD.” Adding further on the importance of using renal dysfunction for SCD “The determination of renal dysfunction as an independent risk factor of SCD is important because patients with moderate-to-severe renal dysfunction were excluded in the studies on which current ICD implantation guidelines are based.” The study ultimately included 1,676 patients who had visited Fujita Health University Hospital for decompensated heart failure On conducting statistical analyses of the gathered data the research team identified two key independent predictors for the risk of SCD following discharge which is already considered in the current guidelines The second was the estimated glomerular filtration rate (eGFR) which is a measure of how well the kidneys can filter blood By adding eGFR as a predictor alongside LVEF regression models could predict SCD more accurately the predictive power of kidney function in this way diminished over time being more effective in the first few months after discharge roughly a quarter of all cases of SCD occurred within three months of discharge the findings of this study underline the importance of considering additional factors when weighing the benefits of using ICD in a patient “Refining the criteria for ICD implantation holds the potential for significantly improving the prevention of SCD and reducing the incidence of complications arising from unnecessary ICD implantations applying an enhanced stratification methodology involving eGFR may yield better clinical outcomes,” remarks Dr these findings may steer the way for the revision of guidelines for the use of ICDs and improve the outcomes for patients with CHF For more information: https://www.fujita-hu.ac.jp/en/index.html Title of original paper: Renal dysfunction is a time-varying risk predictor of sudden cardiac death in heart failure DOI: https://doi.org/10.1002/ehf2.14892 — A new cohort study among 103,642 adults found that current use of cigars was associated with increased risk .. Food and Drug Administration (FDA) approved .. 2024 — Silence Therapeutics presented end-of-treatment data from its Phase 2 ALPACAR-360 study of zerlasiran 2024 — According to a new study being presented at ACC Asia 2024 in Delhi As part of DAIC's continuing Thought Leadership Series this month Editorial Director Melinda Taschetta-Millane sits .. a global leader in cellular and cell-derived therapeutics for the treatment of .. a pioneering AI medical technology company providing physicians with imaging analysis software .. a private company specializing in cell-based therapies for repairing ischemic tissues announced the completion of the first procedure in a patient utilizing its LIBERTY .. a medication initially developed for type 2 diabetes and obesity See All Webinars See All Videos See All Blogs See All Comparison Charts Metrics details Alzheimer’s disease (AD) is the most common form of dementia and is characterized by the accumulation of amyloid β (Aβ) and phosphorylated tau Although there is growing evidence for the anti-neuroinflammatory and neuroprotective effects of the cannabinoid system modulation we analyzed the expression levels of cannabinoid receptor type II (Cnr2/Cb2) in AppNL-G-F/NL-G-F mice and human AD precuneus which is vulnerable to amyloid deposition in AD on neuroinflammation in primary glial cells and neuroinflammation and cognitive impairment in AppNL-G-F/NL-G-F mice The levels of Cnr2/Cb2 were upregulated in microglia isolated from the cerebral cortex of AppNL-G-F/NL-G-F mice CNR2 expression was also increased in RNAs derived from human precuneus with advanced AD pathology Chronic oral administration of JWH 133 significantly ameliorated the cognitive impairment of AppNL-G-F/NL-G-F mice without neuropsychiatric side effects Microglia and astrocyte mRNAs were directly isolated from the mouse cerebral cortex by magnetic-activated cell sorting and the gene expression was determined by quantitative PCR JWH 133 administration significantly decreased reactive astrocyte markers and microglial C1q an inducer for the reactive astrocytes in AppNL-G-F/NL-G-F mice JWH133 administration inhibited the expression of p-STAT3 (signal transducer and activator of transcription 3) in astrocytes in AppNL-G-F/NL-G-F mice JWH 133 administration suppressed dystrophic presynaptic terminals surrounding amyloid plaques stimulation of microglial CB2 ameliorates cognitive dysfunction in AppNL-G-F/NL-G-F mice by controlling astrocyte activation and inducing beneficial neuroinflammation and our study has implications that CB2 may represent an attractive therapeutic target for the treatment of AD and perhaps other neurodegenerative diseases involving neuroinflammation whether CB2 stimulation is beneficial for AD pathologies is still under debate uncovering the alteration of CB2 expression in the precuneus is important to better understand the inflammatory response in amyloid pathologies we first performed a comparative analysis of CB2 gene expression in isolated microglia from AppNL-G-F/NL-G-F mice and the precuneus of the patients with AD we examined whether chronic CB2 receptor stimulation confers neuroprotection and ameliorates cognitive dysfunction in mice with AD the detailed gene expression of isolated glial cells in mice with AD was analyzed A Schematic overview of gene expression analysis of microglia and astrocytes isolated from AppNL-G-F/NL-G-F B Cnr2 gene expression was analyzed using RNA sequencing (RNA-Seq) in isolated microglia (WT: n = 4 and AppNL-G-F/NL-G-F: n = 4) Data are presented as means ± standard error of the mean (SEM) C Quantitative PCR analysis to determine the expression levels of Cnr2 mRNA in glial cells isolated from 2 and 8-month-old AppNL-G-F/NL-G-F mice and WT mice D Human brain samples were selected for analysis based on the Braak staging as follows: control brain (non-AD) defined as Braak stage (senile plaque: SP): 0–A Braak stage (neurofibrillary tangle: NFT): 0–I; mild-AD brain defined as Braak stage (SP): C Braak stage (NFT): III–IV; and advanced AD brain defined as Braak stage (SP): C and Braak stage (NFT): V-VI Schematic overview of the gene expression analysis of CNR2 in the precuneus of non-AD (n = 12) E Quantitative PCR analysis to determine the expression levels of CNR2 mRNA in the precuneus of non-AD (n = 12) These results indicate that CB2 is involved in AD progression and is commonly upregulated in the human AD brain and microglia isolated from AppNL-G-F/NL-G-F mice A A schematic protocol for JWH 133 treatment in primary microglia C Expression levels of mRNAs in JWH 133-treated primary microglia determined by quantitative PCR Relative expression levels for Tnf (B) and Cxcl10 (C) were determined and are presented as means ± SEM D A schematic protocol for microglial condition medium (MCM) treatment of primary astrocytes F Expression levels of mRNAs in MCM-treated primary astrocytes determined by quantitative PCR Relative expression levels for Psmb8 (E) and H2d (F) were determined and are presented as means ± SEM A Experimental timeline for JWH 133 administration and analysis of AppNL-G-F mice B An experimental protocol for the novel object recognition test D Effects of JWH 133 on performance in the novel object recognition test in WT and AppNL-G-F/NL-G-F mice Exploratory preference (%) and time are plotted as means ± SEM [Veh-WT (n = 9) *p < 0.05 and **p < 0.01 (two-way ANOVA) E A schematic protocol for the Barnes maze test one hole indicated as black was designated as the target hole with an escape box A probe test was performed 1 day after the last training session where the escape box was hidden (E middle) TQ: target quadrant; OQ: opposite quadrant; RQ: right quadrant; LQ: left quadrant the target hole (gray) was relocated to the position opposite to the original position 1 day after the probe test G Effects of JWH 133 on the performance of WT and AppNL-G-F/NL-G-F mice in the Barnes maze test Values are presented means ± SEM [Veh-WT (n = 9) *p < 0.05 and **p < 0.01 (training: repeated measures three-way ANOVA H Image showing the travel path recorded for 5 min in the open field test arena J Locomotor activity and anxiety were evaluated in an open field test as the time spent in each zone (I) and total distance traveled (J) These results indicate that JWH 133 ameliorates cognitive impairment in AppNL-G-F/NL-G-F mice without neuropsychiatric side effects A–D Representative immunofluorescent images demonstrating the expression of IBA1 (red C) in the cerebral cortices of Veh-/JWH 133-administered WT (left panels) and AppNL-G-F/NL-G-F mice (right panels) at 12 months old Percentages of the area immunopositive for IBA1 (B) or GFAP (D) are quantified Values are presented as means ± SEM (B and D) Seven sections per mouse were quantified.] *p < 0.05 and **p < 0.01 (two-way ANOVA) (B and D) E Expression levels of mRNAs in microglia isolated from JWH 133-administered WT and AppNL-G-F/NL-G-F mice determined using quantitative PCR Relative expression levels of DAM marker (Cd11c) and complementary marker (C1q) are plotted as the mean ± SEM F Expression levels of mRNAs in astrocytes isolated from JWH 133-administered WT and AppNL-G-F/NL-G-F mice determined using quantitative PCR Relative expression levels of reactive astrocytic markers (H2d and Psmb8) are plotted as means ± SEM H Representative immunofluorescence images demonstrating the expression of GFAP (green) or IBA1 (white) in the cerebral cortices of Veh-WT (top panels) Veh-AppNL-G-F/NL-G-F mice (middle panels) and JWH 133-AppNL-G-F/NL-G-F mice (bottom panels) Arrowheads indicate pSTAT3- and GFAP-positive astrocytes (dashed rectangle areas have been magnified in the far right panels) Scale bars: 50 μm (magnified images) and 25 μm (others) (G) Percentages of pSTAT3 / GFAP cells are quantified Seven sections per mouse were quantified.] *p < 0.05 (Student’s t-test) (H) These results suggest that JWH 133 ameliorates neuroinflammation in AppNL-G-F/NL-G-F mice through microglial CB2 receptor and suppresses polarization of reactive astrocytes by inhibiting the STAT3 pathway A Representative micrographs showing BACE1 (green) and Aβ (red) in the cerebral cortices of Veh- or JWH 133-administered WT or AppNL-G-F/NL-G-F mice C Percentage of the area immunopositive for BACE1 (B) or Aβ (C) in the cerebral cortices of Veh- or JWH 133-administered AppNL-G-F/NL-G-F mice Seven sections per mouse were quantified.] *p < 0.05 and **p < 0.01 (Student’s t-test) D–I Expression levels of mRNAs in the cerebral cortices from JWH 133-administered WT and AppNL-G-F/NL-G-F mice were determined using quantitative PCR Relative expression levels of GABAergic markers (Gabra1 Gabrb2 and Gabrg2) are plotted as means ± SEM (D–F) Relative expression levels of glutamatergic markers (Slc17a7 Slc17a6 and Grin1) are plotted as means ± SEM (G–I) *p < 0.05 and **p < 0.01 (two-way ANOVA) (D–I) J–L Expression levels of mRNAs in the cerebral cortices and glial cells isolated from JWH 133-administered WT and AppNL-G-F/NL-G-F mice determined using quantitative PCR Relative expression levels of Bdnf are plotted as means ± SEM (J: cerebral cortex microglia/astrocyte: Veh-AppNL-G-F/NL-G-F (n = 4) and JWH 133-AppNL-G-F/NL-G-F (n = 3)] *p < 0.05 and **p < 0.01 (two-way ANOVA or Student’s t-test) These results suggest that microglial CB2 stimulation does not affect Aβ clearance These data suggest that microglial CB2 stimulation significantly affects neuronal changes we report that the expression of the CNR2/Cnr2 (CB2) gene was commonly upregulated in the precuneus of patients with AD and in microglia isolated from the cerebral cortices of AppNL-G-F/NL-G-F mice using magnetic-activated cell sorting Sustained stimulation of microglial CB2 ameliorates cognitive dysfunction in AppNL-G-F/NL-G-F mice by inducing beneficial neuroinflammation through controlling astrocyte activation we found that JWH 133 administration ameliorated the neuronal abnormalities in AppNL-G-F/NL-G-F mice such as dystrophic neurites and decreased expression of GABAergic molecules in the cerebral cortex Cnr2/CNR2 is predominantly expressed in microglia and is commonly upregulated in AD mouse models and patients with AD microglial CB2 stimulation may reduce the expression of proinflammatory molecules not only in the microglia but also in the astrocytes of patients with AD to control neuroinflammation CB2 stimulation ameliorated the cognitive impairment without neuropsychiatric side effects such as hyperlocomotion and anxiety JWH 133 binds to CB2 with greater affinity than CB1 and acts as a potent CB2 selective agonist Stimulation of microglial CB2 reduces the release of C1q and induces the release of BDNF from microglia Regulated astrocytic activation ameliorates cognitive dysfunction in AppNL-G-F/NL-G-F mice by protecting neuronal functions The results of the present study indicate that sustained stimulation of microglial CB2 ameliorates cognitive dysfunction in AppNL-G-F/NL-G-F mice by inducing beneficial neuroinflammation by controlling microglia–astrocyte crosstalk CB2 may represent an attractive therapeutic target for treating AD and other neurodegenerative diseases involving neuroinflammation The use of human postmortem brain tissue was approved by the Ethics Committees of Research Institute of Environmental Medicine Nagoya University (approval number #328) and Tokyo Metropolitan Institute (approval number # R21-145) Brain tissue samples for RNA preparation were immediately frozen using liquid nitrogen and stored at − 80 °C before use The homozygous AppNL-G-F/NL-G-F mice obtained by crossbreeding were maintained as inbred lines Age-matched C57BL/6 Jcl mice were used as control All mice were maintained under a standard specific pathogen-free environment (12 h light–dark cycle; 23 ± 1 °C; 50 ± 5% humidity) with free access to food and water throughout the experiments Animals were treated according to the guidelines of the Institutional Animal Care and Use Committee of Nagoya University (approval numbers R240025 and R240026 No difference in the body weight or ingested water was observed between the groups throughout the experiments (Supplementary Fig. S1A, B) except that the experiments were conducted under moderately lit conditions (12 lx) The mice were placed in the center of the arena and allowed to explore the open field (40 cm × 40 cm× 30 cm) for 5 min under moderately lit conditions (100 lx) The open field was divided into an inner zone (30 cm × 30 cm) The movement of the mice was measured using a camera mounted above the open field and their activity was analyzed automatically using TimeOFCR1 software (O’Hara & Co. Measurements included distance and time spent in the inner and outer zones The circular open field was elevated to a height of 75 cm above the floor A black acrylic box (17 × 13 × 7 cm) was placed under one of the holes as an escape box The location of the target hole was consistent for a given mouse and mice within a squad were assigned to the same target hole location across sessions We conducted each trial in a spaced fashion so that all mice within a squad completed a given trial before subsequent trials were run We performed training sessions for 5 days (four trials per session) The mice were individually placed in a white acrylic box (13 × 13 × 13 cm) before the start of each trial the white acrylic box was removed to start the trial Each trial ended when the mouse entered the escape box or after 5 min had elapsed we guided it to the hole manually and allowed it to enter the escape box and remain there for 1 min we measured the latency (sec) to reach the target hole using SMART (Bio Research Center to confirm that the mice learned the location of escape box based on navigation by distal cues in the surrounding environment We measured the time spent in each quadrant (sec) of the visits to the target hole using the software Hole exploration in the target quadrant was defined as the percentage of the visits to the three holes in the target quadrant for total hole visits during the test the escape box was moved to 180o from its previous location and the mice were retrained for 5 days of reversal sessions to determine the new location of the escape box we also measured the latency (sec) to reach the new target using the software the cerebral cortex dissected from mice transcardially perfused with phosphate-buffered saline (PBS) after deep anesthesia was dissociated at 37 °C for 15 min using the Neural Tissue Dissociation Kit-Postnatal Neurons (Miltenyi Biotec Germany) with a gentle MACS Dissociator (Miltenyi Biotec) myelin debris was removed by using Myelin Removal Beads II (Miltenyi Biotec) Purified cells were incubated with anti-CD16/CD32 antibodies (Thermo Fisher Scientific and then incubated with anti-CD11b microBeads (Miltenyi Biotec) to isolate microglia CD11b-positive microglia were isolated via magnetic cell sorting through an LS column (Miltenyi Biotec) we incubated each sample with anti-ACSA2 microBeads (Miltenyi Biotec) to isolate astrocytes ACSA2-positive astrocytes were isolated via magnetic cell sorting through an LS column (Miltenyi Biotec) the mice were deeply anesthetized and perfused intracardially with 4% paraformaldehyde in PBS Twenty-micrometer-thick coronal brain sections were fixed with 4% paraformaldehyde in PBS for 5 min and permeabilized with 0.1% Triton X-100/PBS for 10 min After an incubation in blocking solution (5% goat or donkey serum/PBS) for 1 h the sections were incubated with a combination of the following antibodies: rabbit anti-Iba-1 (#019–19741 or mouse anti-Human Amyloid-β (N) (82E1) (#10323 the sections were incubated with fluorescent-conjugated anti-rabbit or anti-goat IgGs (1:1,000; Thermo Fisher Scientific) or Streptavidin Alexa Fluor™ 546 conjugate (1:500; Thermo Fisher Scientific) at room temperature for 1 h the sections were mounted on slides and analyzed using a confocal microscope (LSM700 Quantitative analysis of microscopic images was performed using ImageJ (National Institutes of Health) Protein extraction was performed as previously described [70] mouse brains were homogenized in 5× volumes of Tris buffer (50 mM Tris–HCl pH7.6 and PhosSTOP phosphatase inhibitor cocktail) with 25 strokes using a potter homogenizer and centrifuged at 200,000 × g for 20 min at 4 °C The resultant supernatant was collected as a brain Tris buffer-soluble (TBS) fraction After the addition of the same amount of 2% Triton X-100/Tris buffer the pellet was homogenized on ice and centrifuged at 200,000 × g for 20 min at 4°C The resultant supernatant was collected as the brain Triton X fraction the same amount of 2% SDS containing Tris buffer was added to the pellet the pellet was incubated for 2 h at 37 °C and centrifuged at 200,000 × g for 20 min at 20 °C The resultant supernatant was collected as the brain SDS fraction the pellet was sonicated with 500 μL of 70% formic acid (WAKO) solution The samples were centrifuged at 200,000 × g for 20 min at 4 °C and the resultant supernatant was evaporated for 2 h The pellet was dissolved in the same volume of DMSO as the brain weight and stored at −80 °C until use as the formic acid-soluble (FA) fraction The BCA assay was performed to estimate the protein concentration Equal amounts of total protein were separated via sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE) and transferred to a polyvinylidene difluoride membrane (Immobilon-P; Merck Millipore The membrane was incubated with a blocking buffer (50 mM Tris-HCl (pH7.4) and 2% (w/v) bovine serum albumin (FUJIFILM Wako) followed by incubation with mouse anti-Aβ (#10323 Japan) or mouse anti-ACTB (#a5441,1:5000; Sigma–Aldrich MO) antibody diluted in the blocking buffer at 4 °C for at least 6 h Bound antibodies were detected using horseradish peroxidase-conjugated secondary antibodies and Immobilon Crescendo Western HRP substrate (Merck Millipore) Images were obtained using LAS-4000 mini (Cytiva Japan) with the equipped software (Multi-Gauge; Cytiva) The gels were stained with SYPRO Ruby Protein Stains (BioRad USA) according to the manufacturer’s protocols Images were acquired using FAS-IV (Nippon Genetics brains from mice at postnatal days 1-2 were dissociated in 0.25% Trypsin and 10 mg/mL Dnase I containing PBS at 37 °C for 10 min Dissociated cells were washed and plated on poly-L-lysine-coated flasks in 10% FBS DMEM (DMEM medium supplemented with 10% FBS and 50 µg /mL streptomycin) (Life Technologies) in a 5% CO2 incubator One to two weeks after seeding the cells dissociated from neonatal mouse brains microglia began to appear on the top of the astrocytic monolayer Microglia were separated from the underlying astrocytic monolayer The resulting astrocyte layer was detached using 0.25% trypsin and replated onto culture plates the cultured astrocytes were used as primary astrocytes in all experiments and maintained in 10% FBS DMEM in a 5% CO2 incubator Microglia were pretreated with 5 µM JWH 133 for 1 h and then stimulated with IFN-γ (10 ng/mL; Peprotech EC Ltd RNA samples were isolated from microglia 6 h after IFN-γ stimulation primary microglia were pretreated with 5 μM JWH 133 or DMSO (vehicle; Veh) for 1 h followed by treatment with lipopolysaccharide (LPS) (50 ng/mL; Sigma) for 24 h The culture medium of primary astrocytes was replaced with Veh / LPS- or JWH 133 / LPS-MCM 24 h after seeding RNA samples were isolated from astrocytes 24 h after MCM treatment and software information supporting the conclusions of this article is included within the article and its Supplementary information 2023 Alzheimer’s disease facts and figures Neuroinflammation in Alzheimer’s disease: microglial signature and their relevance to disease Microglia and neuroinflammation: a pathological perspective Microglial dysfunction in brain aging and Alzheimer’s disease Targeting the endocannabinoid system with cannabinoid receptor agonists: pharmacological strategies and therapeutic possibilities Central side-effects of therapies based on CB1 cannabinoid receptor agonists and antagonists: focus on anxiety and depression The CB(2) receptor and its role as a regulator of inflammation Cannabinoid receptors CB1 and CB2 form functional heteromers in brain CB2 cannabinoid receptor agonist ameliorates Alzheimer-like phenotype in AbetaPP/PS1 mice a cannabinoid CB2 receptor selective compound delays disease progression in a mouse model of amyotrophic lateral sclerosis Prolonged oral cannabinoid administration prevents neuroinflammation lowers beta-amyloid levels and improves cognitive performance in Tg APP 2576 mice Microglial CB2 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cell model A Comprehensive Analysis Identified Hub Genes and Associated Drugs in Alzheimer’s Disease GABA from reactive astrocytes impairs memory in mouse models of Alzheimer’s disease Neuropathological stageing of Alzheimer-related changes Neuropathological diagnostic criteria for Alzheimer’s disease Astroglial major histocompatibility complex class I following immune activation leads to behavioral and neuropathological changes Genetic and animal model analyses reveal the pathogenic role of a novel deletion of RELN in schizophrenia Cognitive and emotional alterations in App knock-in mouse models of Abeta amyloidosis Dimethyl fumarate improves cognitive impairment and neuroinflammation in mice with Alzheimer’s disease Innate immune activation of astrocytes impairs neurodevelopment via upregulation of follistatin-like 1 and interferon-induced transmembrane protein 3 Violating the normality assumption may be the lesser of two evils Download references We thank Center for Animal Research and Education (CARE) Nagoya University for a technical support on animal experiments This work was supported by Grant-in-Aid for Scientific Research JP23K14687 (A.S.) and JP22H04923 (CoBiA) (Y.S S.M.) from the Japan Society for the Promotion of Science (JSPS) JST (Moonshot R&D; Grant Number JPMJMS2024) (K.Y.) JP24wm0425014 (K.Y.) and JP21wm0425019 (Y.S. MHLW Research on rare and intractable diseases Program Grant Number Integrated Research Initiative for Living Well with Dementia (IRIDE) of the Tokyo Metropolitan Institute for Geriatrics and Gerontology IRIDE (Y.S.) and The Hori Sciences & Arts Foundation (A.S. Department of Neuroscience and Pathobiology Research Institute of Environmental Medicine Nagoya University Graduate School of Medicine Medical Interactive Research and Academia Industry Collaboration Center Brain Bank for Aging Research (Neuropathology) Tokyo Metropolitan Geriatric Hospital and Institute of Gerontology United Graduate School of Child Development Nagoya City University Graduate School of Medical Sciences Institute for Glyco-core Research (iGCORE) Center for One Medicine Innovative Translational Research (COMIT) Research Institute for Quantum and Chemical Innovation Institutes of Innovation for Future Society CK and YM performed the experiments with support from OK YS and SM selected and provided the postmortem brain tissues TS and TCS provided the AppNL-G-F mice with critical inputs AS and KY interpreted the data and wrote the manuscript with approval from all authors The authors declare no competing interests The experiments using human brains were approved by the Ethics Committees at Nagoya University (approval number #328) and Tokyo Metropolitan Institute (approval number # R21-145) Informed consent was obtained from their families The experiments using genetically modified mice were approved by the Animal Care and Use Committee and the recombinant DNA experiment committee of Nagoya University (approval numbers R240025 and R240026 All procedures were conducted in accordance with the Declaration of Helsinki Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Download citation DOI: https://doi.org/10.1038/s41419-024-07249-6 Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. Volume 12 - 2018 | https://doi.org/10.3389/fnhum.2018.00148 The stability of the MRI scanner throughout a given study is critical in minimizing hardware-induced variability in the acquired imaging data set which could generate image artifacts and would require the replacement of a major component such as its gradient coil randomly occurring hardware-related noise due to a faulty gradient coil on brain morphometric measures derived from T1-weighted images and resting state networks (RSNs) constructed from resting state functional MRI We also introduced a method to detect and minimize the effect of the noise associated with a faulty gradient coil we assessed the reproducibility of these morphometric measures and RSNs before and after gradient coil replacement Our results showed that gradient coil noise could introduce a large number of voxels exhibiting spurious significant connectivity changes in several RSNs censoring the affected volumes during the analysis could minimize these spurious connectivity changes and could lead to reproducible RSNs even after gradient coil replacement One of these factors is the hardware used to acquire the imaging dataset software and coil architectures need to be considered maintaining the scanner’s stability throughout a given imaging study is very important in minimizing hardware-related variability in the acquired imaging dataset scanner stability ensures that the variability in any MRI-derived measures is mainly due to the population being studied scanner stability guarantees that changes observed over time are driven by disease progression which could necessitate replacement of major components such as the scanner’s gradient coil This major change in hardware could introduce unwanted variability into the imaging dataset Faulty MRI scanner gradient coil could also generate artifacts in the acquired images the degree of which could vary depending on the severity of the problem Large artifacts are easier to detect but low intensity randomly occurring artifacts are difficult to discover Some low intensity gradient-coil-generated noise are barely noticeable but have the potential to significantly affect the outcome of the analysis a commonly used approach for analyzing T1-weighted images we also computed regional GM volume from several regions of interest (ROIs) and other RSN-derived metrics including within-network functional connectivity (WNFC) and similarity measures for test-retest reliability assessment Twenty healthy volunteers (male/female = 15/5) from Nagoya University were recruited for this study The participants’ age ranged from 20 years to 25 years (mean = 22 years All participants had no history of psychiatric or neurological disorder twice before (sessions S1 and S2) and twice after (sessions S3 and S4) the MR gradient coil was replaced The average inter-scan interval was 67.8 days between S1 and S2 Gradient coil noise affected some of the scans in S1 and S2 (pre-replacement) but not scans in S3 and S4 (post-replacement) The study was approved by the Ethical Committee of Nagoya University Graduate School of Medicine with approval number 1014-2 All participants signed a written informed consent before joining the study Magnetic resonance images were acquired using a Siemen’s Magnetom Verio (Siemens, Erlanger, Germany) 3.0T MRI scanner with a 32-channel head coil. For each scanning session, a T1-weighted MR image was acquired using a 3D Magnetization Prepared Rapid Acquisition Gradient Echo (MPRAGE; Siemens; Mugler and Brookeman, 1990) pulse sequence with the following imaging parameters: repetition time (TR)/MPRAGE repetition time = 7.4/2500 ms 192 sagittal slices with a distance factor of 50% and 1-mm thickness acquisition matrix dimension = 256 × 256 and in-plane voxel resolution of 1.0 × 1.0 mm2 Resting state functional MRI scans were also acquired using gradient echo (GE) echo planar imaging (EPI) with the following parameters: TR = 2.5 s 39 transverse slices with a 0.5-mm inter-slice interval and 3-mm thickness 3 × 3 × 3 mm3 voxel resolution and 198 volumes Participants were instructed to close their eyes during the scan but not to fall asleep Other MRI scans were also acquired during the same imaging session however the analysis of these datasets will be reported elsewhere which was then normalized to the Montreal Neuroimaging Institute (MNI) standard space together with the deformation fields from DARTEL were used to normalize the component images to MNI The normalized images were modulated to preserve the amount of signal from each region re-sampled to an isotropic voxel size equal to 2 × 2 × 2 mm3 and smoothed using an 8-mm full-width-at-half-maximum (FWHM) Gaussian filter These preprocessed images were then used in the succeeding analysis To compare differences between sessions at the voxel level a commonly used approach for the analysis of T1-weighted images the preprocessed GM images were entered into a paired sample t-test to identify changes between sessions using SPM12 S4 to compare datasets obtained using the same gradient coil as well as S1 vs S4 to compare datasets obtained from different gradient coils Resulting statistical maps were corrected for multiple comparisons using a family-wise error (FWE) correction rate with p < 0.05 Aside from voxel-wise comparisons to assess differences between sessions in anatomical (T1) images we also estimated other metrics for test-retest reliability assessment These metrics were then entered into an intra-class correlation (ICC) analysis In the above equation, BMS is the between-subject variance, EMS is the condition error variance, and k is the number of conditions (Shrout and Fleiss, 1979) closer to 1) indicating that the between-subject error dominates closer to 0) indicating that the effect of conditions dominates the error For this analysis, we used regional GM volumes computed from the preprocessed GM images from several ROIs defined by the AAL template (Tzourio-Mazoyer et al., 2002) The estimated regional GM volume for each ROI from all participants and sessions were then entered into the ICC analysis Mean signals from selected ROIs within WM and CSF and the global signal plus the temporal difference of these signals The cleaned data were then bandpass filtered within 0.01–0.1 Hz Rotation parameters were converted to displacements along the surface of a sphere with radius equal to 50 mm this value being the approximate mean distance from the cerebral cortex to the center of the head referred to as “noise-corrected” analysis we censored volumes with gradient coil noise before running dual regression analysis An algorithm to detect the gradient coil noise is outlined in “Detection of Gradient Coil Noise” section we performed a one-sample t-test using participant-specific RSN images from all participants in all sessions using SPM12 to generate the mask for the dorsal default mode network (DMN) a one-sample t-test was performed using participant-specific dorsal DMN images from all participants and sessions A threshold value equal to p < 0.0001 corrected for multiple comparisons using FWE correction was then applied to the resulting statistical map and all voxels with p-values below the threshold were included in the mask We used a stringent threshold value to include only voxels with very significant network connectivity the generated mask for that RSN was then applied to the participant-specific RSN image and the mean of the voxel values within the mask was computed and assigned to WNFC The estimated WNFC values from all participants and sessions were then entered into the ICC analysis for test-retest reliability assessments Aside from WNFC, we also assessed the spatial similarity of the participant-specific RSNs relative to the mean RSN image computed over all participants and sessions. We used a spatial similarity measure given by η2 (Cohen et al., 2008; Choe et al., 2015): where ai and bi are values at voxel i in maps a and b mi is the mean value of the two images at voxel i M¯ is the grand mean across the mean image m a is the participant-specific RSN and b is the mean RSN with 0 indicating no similarity and 1 being identical The main advantage of this measure is that it enables the quantification of the similarity or the difference between two images instead of just the correlation between the two images Sample echo planar images from the resting state data of subject 001 (volume 60) 30 and 33 displayed using intensity values ranging from 10 to 1000 (B) Same slices as in (A) but displayed using intensity values ranging from 10 to 50 to highlight the gradient coil noise throughout the image we used the mean intensity of the background image outside the brain we assumed the following: (1) the mean background intensity Ibg outside the brain without gradient coil noise is almost constant throughout the scan and can have different value for each slice; and (2) the mean background intensity with gradient coil noise Inoise is greater than Ibg we generated an individualized outside-brain mask using the outside-brain component obtained from the segmentation of the anatomical image This component was co-registered to the mean functional image to be in the same subject space as the realigned functional images Volumes with at least one slice labeled as noisy were censored in the noise-corrected analyses In order to obtain a reasonable estimate of the individualized RSNs participants whose remaining number of volumes after noise correction was less than 120 volumes (5 min) were excluded Outline of the proposed method for the automatic detection of gradient coil noise All paired sample t-test comparisons of preprocessed GM images did not show any significant difference between sessions using FWE p < 0.05. Estimated ICC values for the 116 ROIs in the AAL template are plotted in Figure 3A The mean ICC value across all ROIs was 0.96 (SD = 0.02) and ICC values ranged from 0.87 to 0.99 All values are close to 1 indicating that between-subject differences dominate compared to the differences among the four sessions This result is reasonable as T1-weighted images appeared to be not affected by the faulty gradient coil Intra-class correlation (ICC) values of regional gray matter (GM) volume within 116 AAL regions of interest (ROIs; A) and estimated η2 values of the 18 resting state networks (RSNs; B) Figure 1 demonstrates the effect of a faulty gradient coil on the echo planar images obtained during sessions S1 and S2. The generated gradient coil noise was not immediately visible when slices were viewed at full intensity range (Figure 1A). However, the noise, characterized by a striped pattern throughout the image, became more evident at low intensity values (Figure 1B) This gradient coil noise appeared randomly sometimes affecting most of the volumes (and slices within the volume) in the scan only affecting a limited number of volumes or slices or none at all Mean background intensity for the entire image (1st row) and per slice (2nd row) showed sharp intensity increases in volumes/slices corrupted by the noise The number of slices considered as noisy for each volume is plotted in the 3rd row with the specific noisy slices shown in the last row A value of 3 was chosen for the threshold in the succeeding analyses since most noisy datasets have plateaued at around this value Performance of the proposed automatic detection method Plots of the detected number of volumes with gradient coil noise as a function of the threshold value ranging from 0.1 to 10 for all datasets Inset: receiver operating characteristic (ROC) curves for two representative datasets (from subject 001 in S1 and subject 002 in S2) affected by gradient coil noise Detection of gradient coil noise using the approach described in the main text for two representative datasets from subject 001 (A) and subject 002 (B) The first row is the mean intensity of all voxels within the individualized outside-brain mask plotted against volume number The second row is the mean intensity per slice The third row is the number of slices affected by the gradient coil noise plotted against volume number as detected by the described method The last row shows the same information per slice (shown in yellow) Number of resting state functional magnetic resonance imaging (fMRI) volumes censored due to head motion (mean FD > 0.2) and gradient coil noise (threshold = 3) Participants’ data excluded in paired sample comparisons for the motion-corrected and scanner-noise-corrected dual regression analyses we identified 18 components related to known RSNs These include primary visual (anterior and posterior) salience (anterior and posterior) and basal ganglia networks Mean and standard deviation (SD) of within-network functional connectivity (WNFC) values across participants for each session and the estimated intra-class correlation (ICC) value Box plots of η2 values for the 18 RSNs are shown in Figure 3B The degree of spatial similarity of the participant-specific RSNs compared with the mean RSN was found to be high with mean η2 values across participants and sessions ranging from 0.73 for the cerebellum (Cer) to 0.79 for the primary visual (anterior Note that there were some participant-specific RSNs with η2 values that were identified as outliers (red plus sign in the plot) particularly in primary visual network (PVis_A and PVis_B) and high visual network (HVis_A) We also observed significant connectivity changes in primary visual and high visual networks between S1 and S3 and in high visual network between S1 and S4 the cerebellum showed significant connectivity changes between S3 and S4 Voxel count for no-correction dual regression analyses Voxel count for motion-corrected dual regression analyses Voxel count for gradient-coil-noise-corrected dual regression analyses Effects of the correction methods in removing the spurious significant connectivity changes between sessions S1 and S2 observed in (A) primary visual network and (B) high visual network (lateral) RSNs are shown in green (group independent components (ICs) z > 3) censoring volumes affected by gradient coil noise could lead to reproducible RSNs on datasets obtained with the same gradient coil (i.e. S4) and between different gradient coils (i.e. simultaneously censoring both gradient coil noise and head motion was not possible since it would result to several datasets with volumes less than 120 we included in Supplementary Table S1 (excluded data sets) and Supplementary Table S2 (voxel count) the case where both gradient coil noise and head motion were simultaneously censored lowering the minimum number of volumes to 96 (4 min) and for a limited data set spurious connectivity changes were significantly reduced in comparisons between sessions within the same gradient coil (S1 vs S2) and across different gradient coils (e.g. S4) suggesting the reproducibility of RSNs before and after gradient coil replacement the authors did not mention the presence of noise in the used resting state fMRI data before gradient coil replacement or considered the effect of noise on the reproducibility of resting state fMRI To mitigate this issue, we introduced a technique which significantly reduced, if not completely removed, spurious connectivity changes driven by the gradient coil noise in some resting state fMRI data. We employed a similar method, called scrubbing or censoring, used to minimize motion artifacts in the presence of large head motion (Power et al., 2015) Application of this method reduced the number of voxels exhibiting spurious connectivity changes in all networks except in high visual and basal ganglia networks The remaining voxels could be just due to other factors including motion artifact The disadvantage of the method is that it reduced the number of volumes included in the analysis and led to the exclusion of the entire dataset for extremely noisy scans a method that would only remove the noise from the image without discarding the volumes in the analysis would be more beneficial Another concern with censoring relates to the differences in the number of volumes included in the analysis (degrees of freedom) for each participant, which could introduce variability in the estimates of the correlation values. In our analysis, we set the minimum number of volumes to 120 (5 min) and datasets with volumes less than this minimum were excluded. As shown by Van Dijk et al. (2010) estimates of correlation strengths started to stabilize with acquisition times as brief as 5 min We can therefore assume that in the preceding analyses the variability introduced due to the differences in the number of volumes would be minimal most of the datasets with censored volumes were excluded and only one dataset (subject 019 in S1) where the number of censored volumes was greater than six was included The findings of these analyses were therefore minimally affected by this issue the number of censored volumes for each participant did vary from session to session the results of these analyses should be carefully interpreted with this potential limitation We also proposed a reliable method to automatically detect the gradient coil noise from the resting state fMRI data The method relied on the noise characteristics observed in our dataset it assumed that the mean intensity of the background image (outside brain) was constant in the absence of gradient coil noise It may also work with other types of randomly occurring image artifacts such as image reconstruction failures as demonstrated in one of the resting state fMRI data its general applicability to other types of noise will depend on the characteristics of the noise being considered As we have mentioned in the results section the observed plateau in the plot could indicate the range of threshold values separating Ibg from Inoise the value where the plot starts to plateau could therefore be used as the appropriate threshold This worked for most of our noisy datasets although its general applicability still remains to be validated hardware-related noise such as those due to a faulty gradient coil could significantly affect the reproducibility of RSNs this type of noise could introduce spurious connectivity changes in several RSNs which could be easily mistaken as valid changes By censoring corrupted image volumes during the analysis the number of spurious connectivity could be significantly minimized if not completely removed Applying this correction method could also make RSNs reproducible even after gradient coil replacement resting state fMRI datasets affected by hardware problems could still be used to generate consistent and reproducible RSNs SN and GS conceived and designed the study SK and HI performed the experiments and analyzed the dataset HW and SM wrote the draft of the manuscript and all authors reviewed and approved the final manuscript This work was supported by Grants-in-Aid from the Research Committee of Central Nervous System Degenerative Diseases by the Ministry of Health Labour and Welfare and from the Integrated Research on Neuropsychiatric Disorders project carried out under the Strategic Research for Brain Sciences by the Ministry of Education This work was also supported by a Grant-in-Aid for Scientific Research from the Ministry of Education Science and Technology (MEXT) of Japan (grant number 80569781) and a Grant-in-Aid for Scientific Research on Innovative Areas (Brain Protein Aging and Dementia Control; 26117002) from MEXT The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fnhum.2018.00148/full#supplementary-material A fast diffeomorphic image registration algorithm Voxel-based morphometry—The methods Beyond feeling: chronic pain hurts the brain disrupting the default-mode network dynamics Functional reorganization of the default mode network across chronic pain conditions Investigations into resting-state connectivity using independent component analysis How reliable are the results from functional magnetic resonance imaging Changes in structural and functional connectivity among resting-state networks across the human lifespan Group independent component analysis reveals consistent resting-state networks across multiple sessions Reproducibility and temporal structure in weekly resting-state fMRI over a period of 3.5 years Defining functional areas in individual human brains using resting functional connectivity MRI Identifying and mapping connectivity patterns of brain network hubs in Alzheimer’s disease Consistent resting-state networks across healthy subjects Modifications of default-mode network connectivity in patients with cerebral glioma Distinct patterns of brain activity in young carriers of the APOE-ε4 allele Clinical applications of resting state functional connectivity The human brain is intrinsically organized into dynamic Resting-state networks in the infant brain Resting-state functional connectivity in neuropsychiatric disorders Functional connectivity in the resting brain: a network analysis of the default mode hypothesis Default-mode network activity distinguishes Alzheimer’s disease from healthy aging: evidence from functional MRI One-year test-retest reliability of intrinsic connectivity network fMRI in older adults Resting state functional connectivity of the striatum in Parkinson’s disease resting-state BOLD and DTI data between identical scanners Resting-state fMRI: a review of methods and clinical applications Altered periaqueductal gray resting state functional connectivity in migraine and the modulation effect of treatment Tutorial on use of intraclass correlation coefficients for assessing intertest reliability and its application in functional near-infrared spectroscopy-based brain imaging Evaluation of resting state networks in patients with gliomas: connectivity changes in the unaffected side and its relation to cognitive function The posterior medial cortex in urologic chronic pelvic pain syndrome: detachment from default mode network-a resting-state study from the MAPP Research Network Three-dimensional magnetization-prepared rapid gradient-echo imaging (3D MP RAGE) Nonparametric permutation tests for functional neuroimaging: a primer with examples Spurious but systematic correlations in functional connectivity MRI networks arise from subject motion characterize and remove motion artifact in resting state fMRI Recent progress and outstanding issues in motion correction in resting state fMRI Reorganization of brain networks in aging: a review of functional connectivity studies Resting-state spontaneous fluctuations in brain activity Decoding subject-driven cognitive states with whole-brain connectivity patterns Intraclass correlations: uses in assessing rater reliability Threshold-free cluster enhancement: addressing problems of smoothing threshold dependence and localisation in cluster inference Longitudinal analysis of neural network development in preterm infants Szewczyk-Krolikowski Functional connectivity in the basal ganglia network differentiates PD patients from controls CrossRef Full Text | Google Scholar Tzourio-Mazoyer Automated anatomical labeling of activations in SPM using a macroscopic anatomical parcellation of the MNI MRI single-subject brain Intrinsic functional connectivity as a tool for human connectomics: theory Neurometrics of intrinsic connectivity networks at rest using fMRI: retest reliability and cross-validation using a meta-level method The default mode network is disrupted in Parkinson’s disease with visual hallucinations Reliable intrinsic connectivity networks: test-retest evaluation using ICA and dual regression approach Naganawa S and Sobue G (2018) Effects of Gradient Coil Noise and Gradient Coil Replacement on the Reproducibility of Resting State Networks Received: 05 August 2017; Accepted: 03 April 2018; Published: 19 April 2018 Copyright © 2018 Bagarinao, Tsuzuki, Yoshida, Ozawa, Kuzuya, Otani, Koyama, Isoda, Watanabe, Maesawa, Naganawa and Sobue. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) *Correspondence: Gen Sobue, c29idWVnQG1lZC5uYWdveWEtdS5hYy5qcA== Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher. 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish. Tiia Olkkonen of Finland celebrates with teammate Vilja Kauranen after the Women's 3 x 5km Relay at the World University Games on Friday in Lake Placid. (Provided photo — Isaiah Vazquez, FISU Games) LAKE PLACID — Finland was a favorite to win the cross-country skiing women’s 3 x 5-kilometer relay at the World University Games. Of the three on the team — Tiia Olkkonen, Vilja Kauranen, and Hilla Niemela — two had won medals earlier in the games. But with several inches of fresh snow on the course, it was not an easy victory for Finns; after a long, hard week of racing, they “were not the freshest ones anymore,” noted Olkkonen. Still, Niemela was able to hold off Norway’s Karianne Olsvik Dengerud, thrusting her ski across the finish line first, 0.7 seconds ahead of Dengerud. “I didn’t think about anything but I have to be the first to the finish line; it’s my job,” said Niemela. “Hilla is my best friend, and we train together,” added Kauranen. “I trusted her, and it was so good to see her come in first.” In the final leg of the 3 x 5km relay, Niemela passed first Norway, then Japan to take the lead in the race. Norway’s Dengerud kept pace with Niemela and Kazakhstan’s Xeniya Shalygina gained ground on Japan’s Rin Sobue, who started the final leg in first. Sobue won a gold medal in the team sprint on the first day of Lake Placid 2023 FISU Games competition a week ago. On the final climb, Niemela surged, and only Dengerud could go with her. Niemela and Dengerud battled up the finish stretch, with the Finnish skier crossing the line for the win in 43 minutes, 49.8 seconds and Dengerud clocking 43 minutes, 50.3 seconds, giving Norway the silver. “I was honestly pretty sure I was going to overtake her,” said Dengerud. “But then it was really hard to pass anywhere in the downhill, with the really heavy snow, and I had a terrible last turn, and then it was hard to get the pace back up. “But I am super happy with the team effort. It’s like a gold for us today and we are just super happy with the race.” Astrid Slav started the relay for the Norwegians and tagged Selma Anderson, who then tagged Dengerud, who won a bronze medal in the cross-country mixed team sprint earlier in the Games. In a surprise move, Japan–which has been dominant in Nordic competition at the Lake Placid 2023 FISU Games–did not make the podium. Instead, Shalygina pulled Kazakhstan from sixth place into podium contention on the final lap of the relay, winning bronze for her team in 42 minutes, 33.2 seconds. Sobue for Japan crossed the line in fourth place. This is Kazakhstan’s first medal at the Lake Placid 2023 FISU World University Games. Aisha Rakisheva and Nadezhda Stepashkina skied the first two legs of the relay. Shalygina, 24, finished fourth in both the women’s individual and pursuit races. She is a 2022 Olympian, three-time national champion for Kazakstan, and competed at the 2022/2023 Tour de Ski earlier this month. She studies at the Kazakh Academy of Sport and Tourism in Almaty. The entire Norwegian women’s relay team studies at American universities: Astrid Stav at the University of Alaska-Anchorage, Delma Anderson at Denver University, and Dengerud at Utah. Cross-country races at Lake Placid 2023 conclude on Sunday at Mount Van Hoevenberg with the women’s and men’s 15k/30k mass starts in the freestyle technique. LAKE PLACID — Registration for the first-ever event on the Adirondack Rail Trail — Sunday, May 4, starting at ... Copyright © 2025 Adirondack Publishing Company | https://www.adirondackdailyenterprise.com | 54 Broadway, Saranac Lake, NY 12983 | 518-891-2600 Neuromuscular Disorders and Peripheral Neuropathies Volume 13 - 2022 | https://doi.org/10.3389/fneur.2022.905613 This article is part of the Research TopicNeuromuscular Disorders and Peripheral Neuropathies – Case Report Collection 2022View all 32 articles and hereditary neurodegenerative disease caused by the testosterone-dependent accumulation of pathogenic polyglutamine-expanded androgen receptor protein two treatments for SBMA have recently been approved in Japan; this decision was based on the results of randomized controlled trials: First anti-androgen therapy using leuprorelin acetate (leuprorelin) a disease-modifying drug that can inhibit the progression of dysphagia but has not yet been proved to improve gait function; second cybernic treatment with a wearable cyborg hybrid assistive limb (HAL®) (Cyberdyne Inc The HAL is an innovative walking exercise system that has been shown to significantly improve gait function in eight neuromuscular diseases without reduction in muscle function It is possible that the combination of these two approaches might yield better outcomes the long-term effects of such a combined approach have yet to be clinically evaluated we describe the case of a 39-year-old male with SBMA who commenced anti-androgen therapy with leuprorelin 1 year previously; this was followed by cybernic treatment with HAL The duration of walking exercise with HAL was 20–30 min a day in one session the patient underwent nine sessions (one course) The efficacy of HAL was evaluated by gait function tests before and after one course of cybernic treatment leuprorelin treatment was combined with cybernic sessions every 2 months for 2 years (13 courses in total) improved by 20.3% in the first course and peaked 10 months after the commencement of combined therapy (a 59.0% improvement) Walking function was maintained throughout the period SBMA is characterized by moderately increased serum levels of creatine kinase (CK) reflecting neuromuscular damage; interestingly the patient's CK levels decreased dramatically with combined therapy indicating remarkable functional improvement Long-term combined therapy improved the patient's gait function with a steady reduction in CK levels The combination of leuprorelin with cybernic treatment can improve and maintain gait function without damaging the motor unit and may also suppress disease progression Over time, SBMA gradually impairs gait function, thus leading to the need for patients to use a wheelchair ~20 years after the onset (6) Bulbar palsy symptoms occur in almost all patients and the most common cause of death is aspiration pneumonia caused by dysphagia the treatment strategy for SBMA is to prevent aspiration and maintain ambulatory function this disease is characterized by a slow progressive deterioration of various motor functions thus resulting in a prolonged and gradually increasing burden on the patient there is a clear need to establish treatments that can alleviate each symptom and potentially cure the disease completely Randomized controlled clinical trials of leuprorelin for the treatment of SBMA ultimately demonstrated the benefit of this drug for the preservation of swallowing function in a patient treatment group (11, 12). In addition, some patients receiving leuprorelin exhibited a significant reduction in the accumulation of mutant ARs in scrotal skin biopsies (11) There was also concern that even if treatment led to an improvement in dysphagia if muscle weakness and walking function did not improve the patient would fall more frequently; this would lead to bone fractures and reduce the patient's quality of life and life expectancy There are cases of patients with SBMA being treated with various exercise therapies, ranging from high to low intensity. However, there have been no reports of improvement in terms of gait function in these cases (16–18) when patients with SBMA are treated with conventional exercise therapy the remaining motor neurons may activate more frequently thus resulting in overexcitement in the motor unit and a likelihood of neuronal loss The HAL controls the torque of the electric power unit based on the wearer's motor unit potentials (CVC) The wearer can walk without overloading the lower limbs the HAL and the wearer can move the lower limbs with HAL's built-in ideal gait pattern based on force plate signals and joint angle measurements (CAC) the wearer senses that the gait patterns close to the ideal have been achieved using CIC These mechanisms allow the wearer to repeat the successful gait in a safe manner without fatigue thus promoting motor learning and the regeneration of walking function Figure 1. A photograph showing a hybrid-assisted limb (HAL) for medical use (lower limb type) (20) we also used a mobile hoist (All-in-One® Denmark) to prevent falls during cybernic treatment with HAL it had been thought that exercise therapy might cause the overexcitement of motor units and exacerbate diseases However, a randomized controlled clinical trial (NCY-3001) demonstrated that innovative HAL-based gait exercise, or cybernic treatment, was safe and efficacious when used to treat eight neuromuscular diseases, including SBMA (20) cybernic treatment was officially approved in Japan and the USA in 2015 and 2020 As with the limitations of leuprorelin therapy when used alone (12, 15), we considered that, without the combined effect of a disease-modifying drug, such as leuprorelin, the effect of HAL alone would not last long enough to inhibit disease progression (20) we hypothesized that the combination of leuprorelin and HAL might overcome the limitations of each of the two therapies alone we focused on the lack of efficacy for leuprorelin when used alone to treat gait disturbance in SBMA We hypothesized that the combination of leuprorelin and HAL would further enhance the efficacy of disease-modifying drugs Japan is the only country in which both therapies are officially approved in a unique position in that they can use leuprorelin and HAL simultaneously to treat patients with SBMA Since conducting a randomized clinical trial of such a combined treatment is not easy we believe that a well-designed observational study like ours is essential and have reported our data here as a case report The patient's serum testosterone levels were quickly suppressed and maintained below the lower limit of the normal range the patient noticed progressive limb weakness indicating the anti-anabolic effect of the anti-androgen therapy the patient received no form of physical therapy One year after the patient's first injection of leuprorelin the patient started intensive and innovative motor learning in the form of cybernic treatment with HAL; this treatment took place in a hospital with specialized rehabilitation facilities for neuromuscular diseases the following neurological findings were noted There was mild muscle weakness in the oris muscles and slight atrophy of the tongue Dysarthria only involved a slight nasal voice; there was no dysphagia The patient had a slight tremor in both upper limbs There was mild weakness in the proximal muscles of the extremities and the tendon reflexes of the extremities were slightly attenuated There was no joint deformity or limitations in the range of motion although he used a cane to walk long distances these data were better than before the onset of HAL treatment The effect of cybernic treatment with a hybrid-assisted limb (HAL) on gait function (A) The black dots indicate the measured walk distance (a 2-min walk test (B) The black dots indicate the speed (a 10-min walk test Vertical gray bars indicate one course of cybernic treatment with HAL this implied that the walking endurance continued to improve with long-term cybernic treatment regular cybernic treatment with HAL appears to improve and maintain walking function The improvement and maintenance of gait function the repeated drastic reduction in serum CK levels and the increase and maintenance of serum creatinine levels were observed only after the combined therapy of HAL and leuprorelin and not during the period of leuprorelin monotherapy (from −13 to 0 months) The effect of cybernic treatment with a hybrid-assisted limb (HAL) on the serum levels of creatine kinase and creatinine (A) The black dots indicate the serum levels of creatine kinase (B) The black dots indicate the serum levels of creatinine This report describes the first successful case in the history of SBMA treatment in which the improvement and maintenance of ambulatory motor function were observed over 2 years we also observed a reduction in serum CK levels as a biomarker of muscle damage and the maintenance of serum creatinine levels as a biomarker of muscle volume retention Our interpretation is that these outcomes are the result of the combination of leuprorelin and HAL Cybernic treatment might improve motor function and inhibit the degeneration of motor neurons Furthermore, this combined treatment led to an improvement of ambulatory function that was maintained for 2 years, with high levels of patient satisfaction. During a previous study of the natural history of SBMA, the 6-min walk test (6MWT) was found to decrease by 11.3% per year (27). The 2MWT is a simplified version of the 6MWT that correlates accurately with a lower patient burden (28); the rate of decrease is expected to be similar We found that the 2MWT at the start of HAL was 111.5 m If we apply a rate of decrease based on the natural history of this disease we can estimate that the 2MWT would have decreased to ~88 m after 2 years the actual value increased to 178.0 m; this was +102% of the estimated value and Leuprorelin was approved in Japan for the treatment of SBMA (13) primarily because of its biological mechanism as a disease-modifying agent that acts on the primary etiology of SBMA and, secondly, because of its proven clinical efficacy for dysphagia in clinical trials (12, 24) Clinical trials of leuprorelin monotherapy did not demonstrate any improvement in ambulatory function In addition to SMBA, similar problems were observed in the lack of improvement in gait function, following the administration of exon-skipping therapy after the onset of DMD (29) and the lack of effect of SMN protein-enhancing treatment in patients with SMA several years after the onset (30) to significantly amplify the efficacies of disease-modifying drugs acting on the primary etiologies of DMD and SMA it is essential that we combine drug therapy and cybernic treatment with HAL This observational single case report clarifies the clinical efficacy of a combination of leuprorelin treatment for dysphagia and HAL treatment for gait disturbance there is a limitation to be considered in that this combination of therapies was not compared statistically to monotherapies it is still unclear whether the combined treatment of leuprorelin and HAL can constantly reduce serum CK levels and maintain ambulatory function over the long term it appears to be crucial that we conduct future RCTs to compare the combined treatment to HAL monotherapy with regard to changes in 2MWT this type of randomized trial may be challenging in a region where both treatments are officially approved A comparative study may be possible in observational cohorts with more patients in Japan the novel form of combined therapy overcame the apparent limitations of disease-modifying drugs acting on primary etiology and improved the gait function of patients with SBMA over the long term Although this is only a single case report it may be possible to improve the treatment efficacy of DMD and SMA The original contributions presented in the study are included in the article/Supplementary Material further inquiries can be directed to the corresponding author The studies involving human participants were reviewed and approved by the Ethics Committee of National Hospital Organization Niigata National Hospital The patients/participants provided their written informed consent to participate in this study Written informed consent was obtained from the individual(s) for the publication of any potentially identifiable images or data included in this article HN drafted and revised the manuscript and was responsible for the acquisition of data MK revised the manuscript and analyzed/interpreted the data GS was responsible for analysis/interpretation of the data and was also involved with the study design and concept All authors contributed to the article and approved the submitted version This research was supported by Grants-in Aid from the Research Committee of CNS Degenerative Diseases (Reference: 20FC1049) Research on Dissemination of Best Practicable Care for Muscle Dystrophy (Reference: 21FC1006) and Research on Policy Planning and Evaluation for Rare and Intractable Diseases Labour and Welfare Sciences Research Grants AH and GS received royalties from Takeda Pharmaceutical Co. and royalties from Takeda Pharmaceutical Co. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher both physiotherapists in National Hospital Organization Niigata National Hospital and Mr Andrew Paul Wood and Charlesworth Author Services for reviewing the manuscript and providing editing support The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fneur.2022.905613/full#supplementary-material Changes in gait speed and endurance in the 2MWT after commencing combined therapy This video clip compares the baseline (an upper row) with a time point 10 months after the commencement of treatment (a lower row) The first 20 s (a left column) and the last 20 s (a right column) of the test can be observed The lower left video (10 months later and the first 20 s in the 2MWT) shows dramatic gait improvement in speed compared to the baseline (the upper left video) The lower right video shows a dramatic improvement in gait endurance compared to the baseline (the upper right video) The patient used a mobile hoist (All-in-One® the same person in the team of evaluators pulls the mobile hoist according to the patient's gait but is careful to never strain the patient's sling and never affect the patient's gait Progressive proximal spinal and bulbar muscular atrophy of late onset Epidemiological survey of X-linked bulbar and spinal muscular atrophy PubMed Abstract | CrossRef Full Text | Google Scholar Elevated creatine kinase and transaminases in asymptomatic SBMA PubMed Abstract | CrossRef Full Text | Google Scholar Biomarkers of spinal and bulbar muscle atrophy (SBMA): acomprehensive review Natural history of spinal and bulbar muscular atrophy (SBMA): a study of 223 Japanese patients Androgen receptor gene mutations in X-linked spinal and bulbar muscular atrophy Widespread nuclear and cytoplasmic accumulation of mutant androgen receptor in SBMA patients Testosterone reduction prevents phenotypic expression in a transgenic mouse model of spinal and bulbar muscular atrophy Leuprorelin rescues polyglutamine-dependent phenotypes in a transgenic mouse model of spinal and bulbar muscular atrophy Phase 2 trial of leuprorelin in patients with spinal and bulbar muscular atrophy Efficacy and safety of leuprorelin acetate for subjects with spinal and bulbar muscular atrophy: pooled analyses of two randomized-controlled trials 13. 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Available online at: https://www.pmda.go.jp/files/000216634.pdf (accessed February 6 Spinal and bulbar muscular atrophy: skeletal muscle pathology in male patients and heterozygous females Biomarker-based analysis of preclinical progression in spinal and bulbar muscular atrophy Efficacy and safety of leuprorelin in patients with spinal and bulbar muscular atrophy (JASMITT study): a multicentre 25. Nakajima T. Nerve function recovery mechanism by the robot suit. Clin Neurosci. (2016) 34:936–7. Available online at: https://scholar.google.com/scholar?cluster=1089814798738094105 clinical trials and the subjective evaluation of patients: the cyborg-type robot HAL and the treatment of functional regeneration in patients with rare incurable neuromuscular diseases in Japan Humans and Devices in Medical Contexts: Case Studies From Japan Google Scholar Walking capacity evaluated by the 6-minute walk test in spinal and bulbar muscular atrophy Two- and 6-minute walk tests assess walking capability equally in neuromuscular diseases Eteplirsen for paediatric patients with Duchenne muscular dystrophy: a pooled-analysis Nusinersen versus sham control in later-onset spinal muscular atrophy Keywords: spinal and bulbar muscular atrophy Sobue G and Nakajima T (2022) The Combined Efficacy of a Two-Year Period of Cybernic Treatment With a Wearable Cyborg Hybrid-Assistive Limb and Leuprorelin Therapy in a Patient With Spinal and Bulbar Muscular Atrophy: A Case Report Received: 27 March 2022; Accepted: 25 May 2022; Published: 24 June 2022 Copyright © 2022 Nakatsuji, Ikeda, Hashizume, Katsuno, Sobue and Nakajima. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) *Correspondence: Takashi Nakajima, bmFrYWppbWEtbWRAbXFiLmJpZ2xvYmUubmUuanA= Metrics details Rheumatoid arthritis (RA) patients often exhibit finger/wrist joint symptoms and reduced grip strength This study aimed to validate grip strength as a measure of frailty in RA patients Subjects were 424 female RA patients (mean age ± standard deviation Frailty was defined as a score of ≥ 8 points on the Kihon Checklist (KCL) Finger/wrist joint symptoms were defined based on tender or swollen joints Associations between frailty and grip strength were determined using receiver operating characteristic (ROC) curve analysis and multivariable logistic regression analysis There were 179 subjects with frailty (42.2%) Multivariable logistic regression analysis revealed that frailty was significantly associated with grip strength independently of finger/wrist joint symptoms cut-off scores of grip strength for frailty in subjects without and with finger/wrist joint symptoms were 17 kg (sensitivity The results of the present study suggest that grip strength in female RA patients is associated with frailty with a cut-off score of 17 kg (equivalent to Cardiovascular Health Study criteria < 18 kg) when RA patients have no finger/wrist joint symptoms when RA patients have finger/wrist joint symptoms it may be considered to reduce the cut-off score of grip strength the extension of health life expectancy is important also for RA patients A major benefit of grip strength is that it is easy to measure in clinical settings Findings from previous studies have highlighted the importance of detecting frailty early and preventing it in order to promote good health outcomes RA patients often exhibit finger/wrist joint symptoms and reduced grip strength this study aimed to validate the use of grip strength as a measure of frailty in RA patients taking into consideration disease activity Since this study aimed to confirm the validity of the cut-off score of grip strength we first targeted females given the small number of male patients in the source population Comorbidities were defined as diseases that were currently or previously treated by other medical specialists and included diabetes mellitus This retrospective study was approved by the Ethics Committees of Nagoya University School of Medicine (2017-0271) Japanese Red Cross Nagoya Daiichi Hospital (2020-451) Japan Community Health Care Organization Kani Tono Hospital (20110901) and Yokkaichi Municipal Hospital (2017-29) We disclosed information pertaining to the study at the cooperating facilities according to the procedure stipulated by the respective Ethics Committees Informed consent was obtained from all subjects The study was conducted in accordance with the World Medical Association of Helsinki ethical principles for medical research involving human subjects Patients’ individual information was anonymized and grip strength with or without finger/wrist joint symptoms The point closest to the upper left corner was identified as the best cut-off point Univariate analysis of variance was performed and estimated marginal means of grip strength were calculated with age and stratifying by frailty and finger/wrist joint symptoms and SPSS version 28.0.0 software (IBM Corp. P < 0.05 was considered statistically significant A total of 630 RA patients consecutively visited our hospitals between June and August 2021. Among these patients, data on clinical characteristics, including scores for grip strength, KCL, and DAS28-CRP were available for 591. Among these 591 patients, 424 were female. Subject characteristics are summarized in Table 1 and the proportions of those with finger/wrist joint symptoms and ever or current malignancy were all significantly different between those with and without frailty we found that finger/wrist joint symptoms were significantly associated with DAS28-CRP and grip strength was significantly associated with HAQ-DI Table 3 shows ORs for frailty based on multivariable logistic regression analyses Since this study aimed to validate grip strength instead of HAQ-DI as a measure of frailty we decided not to include HAQ-DI in the same model as grip strength DAS28-CRP was entered as a variable in Model 1 whereas finger/wrist joint symptoms was entered as a variable in Model 2 (with comorbidities) and Model 3 (without comorbidities) Frailty was significantly associated with age and grip strength (Model 1/Model 2/Model 3) ROC curves for frailty and grip strength (a) without finger/wrist joint symptoms Area under curve and Cut-offs were (a) 0.697 (95% CI: 0.633–0.761) and 17 kg (sensitivity and (b) 0.726 (95% CI: 0.645–0.808) and 14 kg (sensitivity Estimated marginal means of grip strength with age subjects with no frailty and no finger/wrist joint symptoms (n = 171 with frailty and no finger/wrist joint symptoms (n = 103 with no frailty and finger/wrist joint symptoms (n = 74 and with frailty and finger/wrist joint symptoms (n = 76 the proportion of those with frailty was 58.5/26.7% (grip strength < 18 kg / ≥ 18 kg) the proportion of those with finger/wrist joint symptoms was 43.5/27.6% and mean grip strength (SD) was 11.2 (4.5)/23.2 (4.4) kg which were all significantly different between those with grip strength < 18 kg vs those with grip strength ≥ 18 kg the cut-off score of grip strength in RA patients with finger/wrist joint symptoms was 14 kg which was lower than that in those without finger/wrist joint symptoms the upper limit of 95% CI for mean grip strength in subjects with frailty and no finger/wrist joint symptoms was 17.3 kg and that for those with frailty and finger/wrist joint symptoms was 14.7 kg indicating that RA patients with no finger/wrist joint symptoms having a grip strength less than 18 kg or RA patients with finger/wrist joint symptoms having a grip strength less than 15 kg are frailty These findings suggest that grip strength in RA patients reflects frailty regardless of finger/wrist joint symptoms and disease activity The cut-off score of grip strength corresponding to frailty is < 18 kg (equivalent to CHS criteria) when RA patients have no finger/wrist joint symptoms grip strength can be measured more easily than lower limb muscle strength and done in a shorter time than answering the questions of KCL and finger/wrist joint symptoms can be identified immediately it will be important to clarify which RA patients can be evaluated for frailty using grip strength to lower the grip strength cut-off score when the patients have finger/wrist joint symptoms The significance of our present findings is that it reveals the need to reduce the cut-off score of grip strength to predict frailty in RA patients with finger/wrist joint symptoms maintaining grip strength from the perspective of preventing frailty may lead to also aiming for clinical and functional remission which can be the underlying goal of care in RA patients From the perspective of preventing frailty exercise may have a synergistic effect when performed in combination with RA drug therapy Since some RA patients do not have frailty despite having poor grip strength evaluating frailty based only on upper limb function is insufficient upper limb function measurements are simple to perform and may serve as a screening index for evaluating frailty in daily clinical practice we investigated the association between grip strength and frailty in RA patients Frailty was significantly associated with grip strength Measuring grip strength and checking finger/wrist joint symptoms offer a useful and simple way to assess frailty in daily clinical practice Our findings serve as a foundation for the development of methods to detect and screen for frailty The datasets used and/or analysed during the current study available from the corresponding author on reasonable request Kontis, V. et al. 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PLoS ONE 12, e0178383. https://doi.org/10.1371/journal.pone.0178383 (2017) Download references This research was supported by AMED under Grant Number JP21ek0410086 Takako Sashikata for their assistance with information collection Japanese Red Cross Nagoya Daiichi Hospital Japan Community Health Care Organization Kani Tono Hospital National Center for Geriatrics and Gerontology National Hospital Organization Nagoya Medical Center contributed to the conception and design of the study and drafting/revising the manuscript critically for important intellectual content and revising the manuscript critically for important intellectual content All authors read and approved the final manuscript Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Download citation DOI: https://doi.org/10.1038/s41598-022-21533-5 a shareable link is not currently available for this article Sign up for the Nature Briefing newsletter — what matters in science Metrics details plays multiple roles that range from embryonic neuronal migration to spine formation in the adult brain Results from genetic studies have suggested that RELN is associated with the risk of psychiatric disorders We previously identified a novel exonic deletion of RELN in a patient with SCZ High-resolution copy number variation analysis revealed that this deletion included exons 52 to 58 which truncated the RELN in a similar manner to the Reln Orleans mutation (Relnrl-Orl) We examined the clinical features of this patient and confirmed a decreased serum level of RELN To elucidate the pathophysiological role of the exonic deletion of RELN in SCZ we conducted behavioral and neurochemical analyses using heterozygous Relnrl-Orl/+ mice These mice exhibited abnormalities in anxiety and motor learning; the deficits in motor learning were ameliorated by antipsychotics Methamphetamine-induced hyperactivity and dopamine release were significantly reduced in the Relnrl-Orl/+ mice the levels of GABAergic markers were decreased in the brain of these mice our results suggest that the exonic deletion of RELN plays a pathological role implicating functional changes in the dopaminergic and GABAergic systems Relnrl-Orl mice have not yet been well studied which truncated RELN in a similar manner to the Orleans Reln mutation We performed behavioral and neurochemical analyses using heterozygous Relnrl-Orl/+ mice These mice showed abnormalities in anxiety We also found alterations in the dopaminergic and GABAergic neuronal systems These results suggest that the exonic deletion of RELN plays a pathological role The patient (SCZ0782) was a 58-year-old Japanese male with a family history of SCZ His developmental history during early childhood was unremarkable He had a cheerful disposition and was a class representative in primary school he began to show bizarre behavior and violent outbursts against family members he was seen by a psychiatrist for the first time he repeatedly relapsed with severe delusions and hallucinations his delusions and hallucinations remained unchanged despite adequate trials of antipsychotics and his cognitive functioning progressively deteriorated He complained of a feeling of electricity moving through his head and exhibited disorganized speech and behavior At the time of evaluation for this study at the age of 58 years he still had prominent delusions and auditory hallucinations despite being treated with multiple antipsychotics he was considered to have treatment-resistant SCZ and poverty of thought) and repetitive behavior (handwashing and checking) He spent most of his time alone and strongly refused to interact with others such as gait and balance coordination deficits The blood tests of this patient were unremarkable. T1-weighted magnetic resonance imaging showed atrophy of the left cerebral hemisphere, particularly in the frontal and parietal lobes (Supplementary Fig. S1) There were low-intensity areas in the right cerebral peduncle and left basal ganglia His cognitive impairment at the time of the present study was confirmed with the Minimum Mental State Examination (22/30) His scores on neuropsychological tests were generally 1 to 2 standard deviations below the mean of normal controls and were 1 standard deviation below the mean of SCZ patients These results suggested that the truncated RELN of SCZ0782 was not secreted into the extracellular space Behavioral abnormalities in Relnrl-Orl/+ mice The distance moved in the inner (a) and outer (b) sectors and frequency of sector transition between the inner and outer sectors (c) Data represent the mean ± SEM (n = 23 for WT mice; n = 28 for Relnrl-Orl/+ mice) significantly different from WT mice (Student’s t-test) (d–f) Performance in the social interaction test: the habituation phase (d) sociability (e) and social novelty (f) Data represent the mean ± SEM (n = 16 for WT mice; n = 13 for Relnrl-Orl/+ mice) (g and h) Performance in the rotarod test: the training phase (g) and test phase (h) Data represent the mean ± SEM (n = 27 for WT mice; n = 28 for Relnrl-Orl/+ mice) significantly different from WT mice (two-way ANOVA with repeated measures) (i and j) Locomotor activity in the habituation period (i) and METH-induced hyperlocomotion (j) Data represent the mean ± SEM (n = 14 for saline-treated WT mice; n = 20 for saline-treated Relnrl-Orl/+ mice; n = 15 for METH-treated WT mice; n = 21 for METH-treated Relnrl-Orl/+ mice) significantly different from METH-treated WT mice (Tukey’s multiple comparison test) indicating that social approach behaviors were impaired in the Relnrl-Orl/+ mice There were no apparent differences in the performance of Relnrl-Orl/+ mice and WT mice in the elevated plus maze test, Y-maze test, novel object recognition test, and prepulse inhibition (PPI) test (Supplementary Fig. S3a–d) Expression analysis of dopaminergic markers in the NAc and VTA of Relnrl-Orl/+ mice (a and b) TH immunostaining in the NAc (a) and VTA (b) of Relnrl-Orl/+ mice (c–e) mRNA levels of DAT and dopamine receptors in Relnrl-Orl/+ mice Data represent the mean ± SEM (n = 4 for WT mice; n = 4 for Relnrl-Orl/+ mice) (d) D1 receptor (Drd1) mRNA levels in the NAc (e) D2 receptor (Drd2) mRNA levels in the NAc Decreases in GABAA receptor subunits and GAD mRNA levels in the brain of Relnrl-Orl/+ mice The transcripts in WT and Relnrl-Orl/+ mice were quantified by real-time RT-PCR using the 2−ΔΔCT method and normalized to the internal reference housekeeping gene GAPDH Data represent the mean ± SEM (n = 4 for WT mice; n = 3 to 4 for Relnrl-Orl/+ mice) significantly different from WT mice (two-way ANOVA) Dendritic spine analysis in Relnrl-Orl/+ mice Representative images of the spines of cortical pyramidal neurons (a) (Golgi staining; scale bar: 5 µm) Quantitative analysis of spine density (b) spine length (c) spine diameter (d) spine surface area (e) and spine volume (f) of the cortical pyramidal neurons Data represent the mean ± SEM (n = 20 neurons from four WT mice and n = 25 neurons from five Relnrl-Orl/+ mice in b–f) Effects of antipsychotics on impaired rotarod performance in Relnrl-Orl/+ mice (a) Experimental protocol for the pharmacological study with haloperidol or clozapine (b and c) Effects of haloperidol on impaired performance in the training phase (b) and test phase (c) Data represent the mean ± SEM (n = 25 for vehicle-treated WT mice; n = 24 for vehicle-treated Relnrl-Orl/+ mice; n = 22 for haloperidol-treated WT mice; n = 22 for haloperidol-treated Relnrl-Orl/+ mice) significantly different from vehicle-treated WT mice significantly different from vehicle-treated Relnrl-Orl/+ mice (d and e) Effects of clozapine on impaired performance in the training phase (d) and test phase (e) Data represent the mean ± SEM (n = 21 for vehicle-treated WT mice; n = 20 for vehicle-treated Relnrl-Orl/+ mice; n = 22 for clozapine-treated WT mice; n = 22 for clozapine-treated Relnrl-Orl/+ mice) Whether spine abnormalities are observed in the cerebellum should also be examined involving functional changes in the dopaminergic and GABAergic systems we examined the boundaries of the deletion in more detail High-resolution array comparative genomic hybridization (aCGH) was performed using a NimbleGen custom-made fine-tiling array (90-bp probe spacing; NimbleGen USA) targeting the deletion plus the flanking regions on both sides Using genomic DNA extracted from the blood of SCZ0782 aCGH was performed according to the manufacturer’s instructions CNV determinations were made with Nexus Copy Number software v7.5 (BioDiscovery USA) using the Fast Adaptive States Segmentation Technique 2 (FASST2) algorithm The thresholds used to assess copy number loss were set at log2 values of −0.5 The significance threshold p-value was set at 1 × 10−6 and at least five contiguous probes were required for CNV calls Genomic locations are reported in GRCh38/hg38 coordinates We obtained information on the clinical characteristics of the SCZ patient (SCZ0782) with the exonic deletion of RELN from medical records and results of brain imaging and laboratory tests This patient was administered neuropsychological tests This study was approved by the ethics committee of Nagoya University and written informed consent was obtained from the subject The protein concentration of human serum was measured using the Quick Start Bradford Protein Assay (Bio-Rad Laboratories Human serum was solubilized with sodium dodecyl sulfate sample buffer and equal aliquots of protein (45 µg) were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by western blotting using anti-RELN antibody 142 (MAB5366 Images were captured using a LAS-4000 mini (Fujifilm RBRC00063) inbred strains of mice with the BALB/c background were provided by the RIKEN BioResource Center (Tsukuba The strain was maintained in our laboratory Heterozygous reeler (Relnrl-Orl/+) mice were generated by intercrossing Relnrl-Orlr-Orl/+ males and BALB/c females WT BALB/c littermates were used as controls Relnrl-Orl/+ and WT mice were 10 to 15 weeks old when used in the experiments All of the animal protocols were approved by the Animal Care and Use Committee of Nagoya University Graduate School of Medicine; in addition the Principles for the Care and Use of Laboratory Animals which were approved by the Japanese Pharmacological Society and the National Institutes of Health Guide for the Care and Use of Laboratory Animals were followed 40 cm) and an outer area surrounding the inner area The movement of mice was recorded via a camera mounted above the open field Measurements of activity included distance travelled in each area All sessions were conducted under a condition of illumination (15 lx) the test mouse was placed in the chamber and allowed to explore for 10 minutes who had no prior contact with the subject mouse The test mouse was allowed to explore the entire social test box for a 10-minute session a second unfamiliar mouse (stranger 2) was placed in the chamber that had been empty during the sociability test Measurements were taken of the amount of time spent in each zone by the EthoVision automated tracking program (Noldus A zone was defined as the area surrounding the Plexiglas cylinder (diameter of 19 cm) mice were injected with saline or METH (2 mg/kg intraperitoneal injection (i.p.); Sumitomo Dainippon Pharma and locomotor activity was measured for 180 minutes mice were placed individually in a transparent acrylic cage with a black frosted Plexiglas floor (25 × 25 × 20 cm) and locomotor activity was measured every 5 minutes for 60 minutes using digital counters with an infrared sensor (NS-DAS-8; Neuroscience Relnrl-Orl/+ and WT mice were habituated to the test environment for 120 minutes before the measurement of locomotor activity (habituation period) the rotarod test was performed under a moderate light condition (15 lx) mice were placed on a rod rotating at 6 rpm and the time until the mouse falls from the rod was measured If a mouse stayed on the rod until the end of the 2-minute trial Mice were placed on a rod rotating at 12 rpm Each mouse was subjected to six trials per day with a 15-minute inter-trial interval in the training and test phases We calculated the average value of a set of measurements The apparatus was routinely cleaned with water and ethanol following each session We also examined the effects of antipsychotic drugs Mice were administered vehicle (1% carboxymethyl cellulose-saline i.p.) every day 30 minutes before the rotarod test The arms converged in an equilateral triangular central area that measured 4 cm at its longest axis Mice were placed in the central area of the apparatus and allowed to move freely during an 8-minute session The arm entries were sequentially recorded Alternation was defined as multiple entries into three different arms (A The percent alternation was calculated as the number of alternations divided by the number of total arm entries minus 2 multiplied by 100 defined as successive entries into the three arms on overlapping triplet sets was associated with the capacity of short-term memory The apparatus was made of plastic material and was elevated to a height of 50 cm above the ground Each arm of the plus maze was 16 cm in length and 10 cm in width the closed arms had wall enclosures that were 20 cm high The central platform was a square measuring 10 × 10 cm The light intensity around the maze was set at 100 to 120 lx Mice were placed on the elevated plus maze for 5 minutes They were placed on the elevated plus maze facing the open arm opposite to the experimenter The number of entries and time spent in the open and closed arms were recorded over the entire duration of the test mice were placed in an apparatus (30 × 30 × 35 cm) for 3 days two novel objects were placed in the apparatus and the mice were allowed to explore each object for 10 minutes under a moderate light condition (15 lx) The time spent exploring each object was recorded The preference index was calculated as the ratio of time spent exploring one of the objects to the total exploration time mice were placed back into the same apparatus 24 hours after the training session one of the familiar objects used during training was replaced by a novel object and the mice were allowed to explore each object (familiar and novel object) for 5 minutes The preference index in the test session was defined as the ratio of the amount of time spent exploring the novel object over the total time spent exploring the two objects After mice were placed in the chamber under a moderately bright light condition (180 lx; San Diego Instruments they were habituated for 10 minutes in the presence of background white noise (65 dB) The mice received three kinds of trials (10 startle trials The inter-trial interval ranged between 10 and 20 seconds The startle trial included a single 120-dB white noise burst lasting 40 milliseconds The PPI trials consisted of a prepulse (20-millisecond burst of white noise at an intensity of 69 mice received sixty different trials pseudo-randomly ensuring that each trial was carried out 10 times and that no two consecutive trials were identical We measured the resulting movement of the animal in the startle chamber for 100 milliseconds after startle stimulus onset (sampling frequency which calculated the maximal response over the 100-millisecond period The basal startle amplitude was defined as the mean amplitude of 10 startle trials The PPI was defined as 100 × [1 − (PPx/P120)]% in which PPx was the mean of 10 PPI trials (each of the four prepulse trials; PP69 or PP80) and P120 was the basal startle amplitude Relnrl-Orl/+ and WT mice were deeply anesthetized with tribromoethanol (Avertin; 200 mg/kg i.p.) and transcardially perfused with ice-cold 0.1 M phosphate buffer (pH 7.4) followed by 4% paraformaldehyde in phosphate buffer Their brains were removed and incubated with 4% paraformaldehyde in phosphate buffer overnight brains were incubated with 20% and 30% sucrose in phosphate buffer for 1 day each at 4 °C Brains were cut into 20-µm-thick coronal sections on a cryostat (CM3000; Leica Microsystems GmbH and free-floating sections were used for TH immunohistochemistry Brain slices were incubated with blocking solution (5% normal goat serum and 0.3% Triton X-100 in phosphate-buffered saline) for 30 minutes then incubated with a mouse anti-TH antibody (1:200 Germany) at 4 °C for 24 hours with constant shaking Sections were incubated with peroxidase-labeled polymer at room temperature for 1 hour and the reaction was visualized using 3,3′-diaminobenzidine (EnVision Kit Brains were cut into 80-µm-thick coronal sections on a cryostat (CM3000; Leica Microsystems GmbH The images of pyramidal neurons located in layer II/III of the mPFC were obtained using bright-field microscopic (Keyence Only fully impregnated neurons displaying dendritic trees without obvious truncations and isolated from neighboring impregnated neurons were retained for analysis The quantification of spine density was limited to dendrites 50 to 200 µm from the soma Spine density was calculated as the number of spines per 10 µm of dendrite length All dendrites and spines within images were traced using Neurolucida software (MicroBrightField Bioscience USA) and analyzed using NeuroExplorer (MicroBrightField) Mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal reference housekeeping gene All data are expressed as the mean ± standard error of the mean (SEM) or three-way ANOVA with or without repeated measures was used followed by Tukey’s test when F ratios were significant (p < 0.05) The statistical significance of differences between two groups was assessed using the Student’s t-test Ogawa, M. et al. 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Methods 25, 402–408, https://doi.org/10.1006/meth.2001.1262 (2001) Download references This research was supported by research grants from the Ministry of Education Science and Technology of Japan; the Ministry of Health Labour and Welfare of Japan; the Japan Agency for Medical Research and Development (AMED) under grants No and JP18mk0101076; the Scientific Research on Innovative Areas “Glial assembly: a new regulatory machinery of brain function and disorders” program; the Scientific Research on Innovative Areas “Comprehensive Brain Science Network” program; and the SENSHIN Medical Research Foundation; Uehara Memorial Foundation Taku Nagai and Wei Shan contributed equally Department of Neuropsychopharmacology and Hospital Pharmacy Graduate School of Pharmaceutical Sciences Center for Advanced Medicine and Clinical Research Advanced Diagnostic System Research Laboratory Fujita Health University wrote the main text and prepared most of the figures; A.S. performed most of the behavioral experiments and neurochemical analyses in mice; I.K performed the aCGH experiments and phenotype analysis in the patient; T.K performed western blotting and cell biological experiments; Y.A examined the effects of antipsychotics on impaired rotarod performance in Relnrl-Orl/+ mice; N.K prepared the neuropsychological assessment session; all other authors commented on and refined the manuscript; N.O. All authors have carefully read the paper and approved the final manuscript Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Download citation DOI: https://doi.org/10.1038/s41598-018-31390-w Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily. The painting is the centrepiece of an exhibition to mark the 200th anniversary of what is said to be the world’s most famous poem: Wordsworth’s Daffodils I would like to be emailed about offers, events and updates from The Independent. Read our Privacy notice The first new portrait of William Wordsworth in two centuries years – depicting him much younger than in previous likenesses – is to go on display after a Japanese artist found a “life mask” made years before the poet became a Victorian celebrity. The painting forms the centrepiece of an exhibition by Hideyuki Sobue which opens to mark the 200th anniversary of what is said to be the world’s most famous poem: Wordsworth’s “Daffodils”. Sobue’s diptych features the first new painting of Wordsworth since he died, aged 80, in April 1850. It is based on a mask made by the artist Benjamin Robert Haydon using moulds of the poet’s face in June 1815 – the same year that the final version of “Daffodils” was published. Most of the surviving images of Wordsworth show him in later life. But Sobue’s portrait captures him at the peak of his physical powers, when he was a keen gardener and walker who routinely trod 20 miles to Keswick for lunch and then walked back again. “Most of the portraits are romanticised, but I wanted to capture the emotion of the poet, knowing that he was a keen outdoors man,” said Sobue, who has been living and working in the Lake District since 2005. “I was not satisfied with the images I saw. But when I came across the mask in my researches I was ecstatic.” Sobue’s technique uses sumi ink, traditionally used in East Asian calligraphy, together with acrylic paints on canvas. He builds the images line by line. The work is being shown at Rydal Mount, the house near Ambleside in Cumbria where Wordsworth lived for most of his life, and where he published the definitive version of “Daffodils”. The mask lies beneath the oriel window of the Upper Library in St John’s College, at the University of Cambridge. But Sobue came across it in the National Portrait Gallery’s online archive. The centuries-old tradition of making life and death masks from moulds, capturing a person’s likeness for posterity, expanded in the 19th century thanks to the growing interest in phrenology. According to Haydon’s diary, Wordsworth bore “like a philosopher” the trial of sitting with his face covered with plaster, and with straws up his nose to allow breathing, while the mould was made. Sobue is a great admirer of Wordsworth’s work. He and the poet Gary Boswell worked on a collaborative project at Rydal Mount 10 years ago, in an attempt “to trace back to the poet’s spirituality and creativity from the perspective of our contemporary society”. The new exhibition takes the “Daffodils” poem and the opening words “I wandered” as its inspiration. The staff for Fureru., the new original anime film from the Super Peace Busters creative team began streaming the film's second full trailer on Wednesday The story follows three childhood friends who are connected by a mysterious creature. The film shows the three as young adults in Tokyo. Thee movie is set in Takadanobaba, a different location from the team's previous Chichibu setting of the Super Peace Busters trio's previous three works (Chichibu is Mari Okada's hometown) The film features nine cast members from the team's previous three anime works including anohana: The Flower We Saw That Day, The Anthem of the Heart, and Her Blue Sky The Super Peace Busters creative team consists of director Tatsuyuki Nagai, writer and director Mari Okada, and artist Masayoshi Tanaka. Nagai (anohana: The Flower We Saw That Day, The Anthem of the Heart, and Her Blue Sky) is directing the film at CloverWorks YOASOBI performs the theme song "Monotone." Aniplex is producing the film with Story Inc. and distributing it with TOHO Sources: Aniplex's YouTube channel, Comic Natalie 2018 at 9:00 am ET.css-79elbk{position:relative;}AVON CT — Here are some recent notable accomplishments by local college students To sign up for Avon breaking news alerts and more, click here. Get more local news delivered straight to your inbox. Sign up for free Patch newsletters and alerts. At the AnimeJapan 2024 stage event for the upcoming anime film Fureru. the Super Peace Busters creative team revealed three new character designs for the film’s lead characters The film is set to premiere in Japanese theaters sometime later in Fall 2024 The new designs all showcase the main characters as close childhood friends in their 20s and their individual personalities Ryo Sobue is a first-time real estate agent who is an active and sporty older brother type to his friends Aki Onoda is a part-timer at a bar who has trouble speaking and tends to act first before saying anything Yuta Inohara is a designer and fashion school student who suffers from many unlucky things in his life tending to act very passive because of them reunites members of the creative collective Super Peace Busters which includes Tatsuyuki Nagai (A Certain Scientific Railgun T) as director Dragon Pilot: Hisone & Masotan) as screenwriter CloverWorks is the main animation production company The story is set in modern Tokyo and follows three young men who find a strange creature Source: Fureru. Official Twitter, Anime News Network The official website for Fureru., the new original anime film from the Super Peace Busters creative team revealed on Sunday that nine cast members from the team's previous three anime works will take part in the film The cast members (and the characters they played in the previous anime works) are (from top left to bottom right): Sources: Fureru. anime film's website, Comic Natalie The APRSAF/SAFE Executive Board Meeting was held on November 25 Shashikant Sharma (Indian Space Research Organisation (ISRO)) and Dr Shinichi Sobue (Japan Aerospace Exploration Agency (JAXA)) The main topic of meeting was discussing on progress and future plan of three SAFE projects and SAFE CH4Rice (Assessment of Methane Emission from Rice Paddies and Water Management) Project The participants confirmed the good progress of the two ongoing SAFE projects (Rice Crop Monitoring led by GISTDA and CH4Rice led by VNSC) and also confirmed the successful completion of the Agromet project at the APRSAF-30/SAWG on November 26 2024 and its achievements including its contribution to regional food security through the provision of validated satellite-derived agromet data to ASEAN countries with capacity building events It is expected to expand this activity to South Asia under ISRO-JAXA cooperation and other framework further discussions and collaborations between the public and private sector stakeholders on carbon credit training and IoT devices for water-level and CH4 flux in-situ measurements was outlined Zoom » Agenda and Presentation materials Earth Observation Research Center, Japan Aerospace Exploration Agency Copyright © JAXA EORC All rights reserved Refrigerators are indispensable appliances in our daily lives. In order to add enjoyment and attachment to such refrigerators, ANTBY, a Japanese home appliance manufacturer, has invited poet Toshi Sugawara as a producer to create a new series of "Refrigerator Showcase," which will be held on March 25, 2012.TOBIRAE Focusing on the smallest type of refrigerator we collaborated with photographer Naoki Ishikawa the result is not just an inorganic appliance but an art piece that can be displayed in a room The lineup includes not only refrigerators that can actually be used but also refrigerator speakers that have been upcycled from unusable refrigerators with musician Purdon Kimura REIZOKO SPEAKER Production by PARDON KIMURA in conjunction with the launch of the project Ginza Tsutaya on the 6th floor of GINZA SIX is currently holding a fair with refrigerators from the project Please come and open the door to "Tobirae," a stylish embodiment of the cross-industrial collaboration of home appliances and art Text_Hiroshi Yamamoto The story follows three childhood friends who are connected by a mysterious creature. The film shows the three as young adults in Tokyo. The movie is set in Takadanobaba, a different location from the Super Peace Busters trio's previous three works (Chichibu is Mari Okada's hometown) YOASOBI performs the theme song "Monotone." Sources: Fureru. official website, Comic Natalie The official website for Fureru., the new original anime film from the Super Peace Busters creative team, unveiled a new trailer, visual, cast, and October 4 opening on Friday. The video reveals and previews the theme song "Monotone" by YOASOBI Sources: Fureru. anime film's website, Comic Natalie Japan'smassive Kaguya lunar orbiter stands poised to launch spaceward this week on amission that Equippedwith a veritable arsenal of science instruments and two baby satellites thethree-ton moon probe is set to liftoff from Japan early Friday (Local Time) ona one-year mission to Earth's nearest neighbor "TheJapanese people are very interested in this mission," said Shinichi Sobue,Kaguya's science coordinator and public outreach for the Japan AerospaceExploration Agency (JAXA) is our first mission for reallyobserving the moon." Japan launcheda previous lunar mission in 1990 but the flight served primarily as atechnology demonstrator That mission -- dubbedMuses-A -- sent the Hiten spacecraft on a series of lunar flybys and orbits releasedthe small microsatellite Hagoromo and intentionally crashed into the moon'ssurface in 1993 Kaguya's SELenologicaland ENgineering Explorer (SELENE) mission, meanwhile, is designed for in-depth lunarstudy The probe is slated to lift off atop an H-2A rocket from Japan'sTanegashima Space Center on Sept 14 GMT),though it will be Friday morning at the island launch site Touted byJAXA as the largest lunar mission since NASA's manned Apollo flights Kaguya isnamed after a moon princess in a Japanese folktale and carries 14 primaryscience instruments to map the lunar surface and study its composition it is hoped that we can get closer to the core ofthe mystery of the origin and evolution of the moon," Kaguya's SELENEproject manager Yoshisada Takizawa has said in a JAXA Web site statement Otherinstrument suites will study mineral distribution on the moon's surface; usecameras radar and lasers to catalogue lunar terrain and subsurface structure; andprobe the moon's ionosphere and magnetic field A high-definition camera is alsolaunching aboard Kaguya but is destined for a more aesthetic purpose "TheJapanese people would like to see the very beautiful high-definition movies ofthe Earth rising" over the moon Atop theorbiter sit the relay(RSAT) and VRAD microsatellites two solar-powered probes about three feet(one meter) in diameter Kaguya will jettison the 110-pound (50-kilogram) probesas it enters lunar orbit The two satellites are designed to then work togetherwith their mothership to generate a complete global map of the moon'sgravitational field VRAD will also probe the moon's thin ionosphere "It'sthe first [mission] ever to study the gravity field of the far side of themoon," Sobue said of Kaguya's mission "The data accumulated bySELENE should serve as a basis for mankind's future utilization of the moon." JAXAmission scientists already plan to share Kaguya's gravity field measurementswith NASA researchers as the U.S space agency prepares to launch its ownmission - the Lunar Reconnaissance Orbiter (LRO) - in 2008 and returnastronauts to the moon by 2020 too,hopes to work with international partners to develop a lunar base as part theagency's strategic vision 2025 "Thus,Kaguya will also contribute to the manned exploration of the moon,"Takizawa stated Kaguya is expected to orbit Earth about 2.5 times before beginning thefive-day trek to the moon Thespacecraft is designed to enter into a polar lunar orbit and then deploy the RSATand VRAD microsatellite before ultimately settling into a 62-mile(100-kilometer) high orbit around the moon The probe's one-year science missionshould begin in earnest after a three-month checkout period Kaguya isthe first of series of new spacecraft bound for the moon over the next twoyears China plansto launch its firstmoon shot Chang'e-1 sometime later this year with NASA's LRO and India'sChandrayaan-1 spacecraftset to fly in 2008 Japan also plans to follow Kaguya's flight with more ambitiousmissions under the SELENE banner "[W]ewill plan to have SELENE follow-on missions for moon landing and samplereturn," Sobue said Join our Space Forums to keep talking space on the latest missions, night sky and more! And if you have a news tip, correction or comment, let us know at: community@space.com. Meta Quest 3S review: Fantastic VR performance at a budget price A giant crater on the moon may hold remnants of an ancient magma ocean Artemis astronauts could bring home samples of it a novelist forced to live a bitterly frugal life even as he aims to win the prestigious Akutagawa Prize for literature Oguri is seen in costume in the photograph below right alongside Yurina Hirate as protagonist Hibiki Akui Principal photography for the film began on June 1 The film will open in Japan on September 14 Shō Tsukikawa (live-action Kurosaki-kun no Iinari ni Nante Naranai, I Want to Eat Your Pancreas) is directing the film, with a script by Masafumi Nishida (Tiger & Bunny) a handwritten novel manuscript is sent into a rookie contest but it doesn't meet the conditions and is thrown in the trash an editor named Hanai picks it up by chance and believes it is a revolutionary novel but there is no contact address on the submission Hibiki – a 15-year-old student – joins her school's literary club not knowing that Hanai is trying to track her down Yanamoto launched the manga in Big Comic Superior in August 2014. Shogakukan released the manga's ninth volume on April 27. The manga won the 10th Manga Taisho award last year Source: Comic Natalie