3D multiple immunoimaging using whole male organs in rice

Spatiotemporal regulation of proteins and RNAs is essential for the precise development of reproductive tissues in many organisms. The anther, a prominent part of the male reproductive organ in plants, contains several somatic cell layers named the anther wall and, within it, the germ cells. Here, we successfully developed a simple 3D organ-immunoimaging technique for rice anthers, which distinguishes each individual cell from the four somatic cell layers and germ cells without the need for transformation, embedding, sectioning, or clearing. The 3D immunostaining method is also applicable to the intracellular localization of meiosis-specific proteins in meiocytes, as exemplified by MEL1, a germ cell-specific ARGONAUTE in the cytoplasm, and ZEP1, a pachytene marker on meiotic chromosomes. Our 3D multiple immunostaining method with single-cell and intracellular resolution will contribute to a comprehensive organ-level elucidation of molecular mechanisms and cellular connectivity

miR2118-dependent U-rich phasiRNA production in rice anther wall development

Reproduction-specific small RNAs are vital regulators of germline development in animals and plants. MicroRNA2118 (miR2118) is conserved in plants and induces the production of phased small interfering RNAs (phasiRNAs). To reveal the biological functions of miR2118, we describe here rice mutants with large deletions of the miR2118 cluster. Our results demonstrate that the loss of miR2118 causes severe male and female sterility in rice, associated with marked morphological and developmental abnormalities in somatic anther wall cells. Small RNA profiling reveals that miR2118-dependent 21-nucleotide (nt) phasiRNAs in the anther wall are U-rich, distinct from the phasiRNAs in germ cells. Furthermore, the miR2118-dependent biogenesis of 21-nt phasiRNAs may involve the Argonaute proteins OsAGO1b/OsAGO1d, which are abundant in anther wall cell layers. Our study highlights the site-specific differences of phasiRNAs between somatic anther wall and germ cells, and demonstrates the significance of miR2118/U-phasiRNA functions in anther wall development and rice reproduction

商業施設で行う健康事業としての「健幸大学」への 参加が高齢者の健康に与える影響

本研究は，商業施設で行う健康事業としての「健幸大学」への参加が高齢者の健康にどのような影響を与えたのか明らかにした。「健幸大学」は商業施設を活動の場とし，身体機能測定・講話・レクリエーションを中心としたプログラムで構成し，2019年度の活動は月1～2 回のペースで合計8回実施した。今回,「健幸大学」に複数回参加した高齢者にアンケートを実施し，参加するようになってから①健康への取り組みにどのような変化があったのか，②具体的な気持ちや行動の変化，に関して自由記載で回答を求めた。アンケート回収率81.5％，有効回答数は65 名（有効回答率86.6％）であった。得られた自由記載を，テキストマイニングを用いて単語頻度分析と共起分析を行った。分析の結果，総抽出語数は182 語で，単語頻度分析では，高頻度順に『運動』・『参加』（17 回），『身体』・『良い』（13回），『気を付ける』・『毎日』（11 回），『健康』・『自分』・『少しずつ』（10 回）と続いた。また，共起分析では，3 つのネットワークが形成された。日常生活と関連性が強い買い物が行える商業施設で行う健康事業は，高齢者にとって気軽に，そして繰り返し参加しやすい環境であった。このことが知識獲得の機会となり，複数回参加した高齢者に健康への関心の高まりと健康づくりに対する行動変容が少しずつ起き始めていた。今後は，地域との連携を含めた高齢者の健康行動が習慣化される仕掛けづくりを検討していく

認知症高齢者に対する「聞き書き」による看護学生の実習での学び

本研究の目的は，学生が実施している認知症高齢者への「聞き書き」の学びを明らかにすることである。A大学老年看護学実習において学生が行った認知症高齢者に対する「聞き書き」のリフレクションレポートを質的帰納的に分析した。その結果，【語り手の個人史や価値観の理解が深まる】【聞き書きは個別性のあるケアを創り出す】【語り手の豊かな感情を呼び起こす】【語ることで人生を振りかえり，過去の記憶や思い出に親しむ】【語りを引き出すための事前準備の重要性】【語りを傾聴し，ありのままを受けとめる姿勢の重要性】【記憶の想起を助ける工夫の重要性】【聞き書き中は語り手に合わせた状況判断が求められる】の8つのカテゴリが抽出された。学生は「聞き書き」を通して，認知症高齢者がもつコミュニケーション障害を考え，学生自らが相手に応じて瞬時に状況判断を行って工夫し，認知症高齢者との関係づくりに取り組めていたと考える。また学生は，「聞き書き」を通して，認知症高齢者に対する態度形成とQOLを高めるケア提供の機会を得ていたと考える。これらのことから，「聞き書き」は，学生が認知症高齢者との対話を通して，老年看護学教育において重要な認知症高齢者の理解が深まる機会になることが示唆された。教員は，今後も学生個々が「聞き書き」という相互行為を通して体験した内容，感情，思考などを言語化したものから，学生が認知症高齢者理解を深めていく過程を確認していくことが重要である

Mild Electrical Stimulation with Heat Shock Ameliorates Insulin Resistance via Enhanced Insulin Signaling

Low-intensity electrical current (or mild electrical stimulation; MES) influences signal transduction and activates phosphatidylinositol-3 kinase (PI3K)/Akt pathway. Because insulin resistance is characterized by a marked reduction in insulin-stimulated PI3K-mediated activation of Akt, we asked whether MES could increase Akt phosphorylation and ameliorate insulin resistance. In addition, it was also previously reported that heat shock protein 72 (Hsp72) alleviates hyperglycemia. Thus, we applied MES in combination with heat shock (HS) to in vitro and in vivo models of insulin resistance. Here we show that 10-min treatment with MES at 5 V (0.1 ms pulse duration) together with HS at 42°C increased the phosphorylation of insulin signaling molecules such as insulin receptor substrate (IRS) and Akt in HepG2 cells maintained in high-glucose medium. MES (12 V)+mild HS treatment of high fat-fed mice also increased the phosphorylation of insulin receptor β subunit (IRβ) and Akt in mice liver. In high fat-fed mice and db/db mice, MES+HS treatment for 10 min applied twice a week for 12–15 weeks significantly decreased fasting blood glucose and insulin levels and improved insulin sensitivity. The treated mice showed significantly lower weight of visceral and subcutaneous fat, a markedly improved fatty liver and decreased size of adipocytes. Our findings indicated that the combination of MES and HS alleviated insulin resistance and improved fat metabolism in diabetes mouse models, in part, by enhancing the insulin signaling pathway

Cytoplasmic Fragment of Alcadein alpha Generated by Regulated Intramembrane Proteolysis Enhances Amyloid beta-Protein Precursor (APP) Transport into the Late Secretory Pathway and Facilitates APP Cleavage

The neural type I membrane protein Alcadein alpha (Alc alpha), is primarily cleaved by amyloid beta-protein precursor (APP) alpha-secretase to generate a membrane-associated carboxyl-terminal fragment (Alc alpha CTF), which is further cleaved by gamma-secretase to secrete p3-Alc alpha peptides and generate an intracellular cytoplasmic domain fragment (Alc alpha ICD) in the late secretory pathway. By association with the neural adaptor protein X11L (X11-like), Alc alpha and APP form a ternary complex that suppresses the cleavage of both Alc alpha and APP by regulating the transport of these membrane proteins into the late secretory pathway where secretases are active. However, it has not been revealed how Alc alpha and APP are directed from the ternary complex formed largely in the Golgi into the late secretory pathway to reach a nerve terminus. Using a novel transgenic mouse line expressing excess amounts of human Alc alpha CTF (hAlc alpha CTF) in neurons, we found that expression of hAlc alpha CTF induced excess production of hAlc alpha ICD, which facilitated APP transport into the nerve terminus and enhanced APP metabolism, including A beta generation. In vitro cell studies also demonstrated that excess expression of Alc alpha ICD released both APP and Alc alpha from the ternary complex. These results indicate that regulated intramembrane proteolysis of Alc alpha by gamma-secretase regulates APP trafficking and the production of A beta in vivo

Stabilization of intracellular trafficking and metabolism of amyloid β-protein precursor and Alcadein β by apolipoprotein E

Intracellular metabolism of amyloid beta-protein precursor (APP) is important for the pathogenesis of Alzheimer's disease (AD). Alcadeins (Alc alpha, Alc beta, and Alc gamma) are neural membrane proteins similar to APP in their localization, metabolism, and cellular function. Isoform epsilon 4 of apolipoprotein E (ApoE) is a major risk factor for AD. We found that ApoE expression attenuated intracellular trafficking of APP and Alc beta, resulting in metabolic stabilization of both proteins. By contrast, Alc alpha intracellular proteolysis was facilitated by ApoE expression, which was not due to an increase in the primary cleavage of Alc alpha. This difference may result from binding of ApoE to membrane proteins