野村 純

type:text [要約] 学校管理下でけがが発生した場合，適切な対応を行うために養護教諭は教員との情報共有が重要と考えているが，実際は，教員との連携がうまくいかないと感じる事例が多くある。本報告では「教員の情報共有の意識調査」と「養護教諭の情報共有の実態調査」を行い，教員と養護教諭が行う情報共有の認識と実践の実態を分析した。この結果，けが対応時の情報共有の必要事項に関して両者の間でズレがあることが明らかになった。また，教員の情報共有を行っているという意識と養護教諭からみた教員の情報共有の実態のズレも明らかになった。さらに，けが対応時に情報共有を行っていない教員の中に「情報伝達の意識はあるが，実際は行動につながっていない教員」と「情報伝達の意識が低く，行動につながらなかった教員」がいる実態も判明した。

type:text [要約] 平成27年中央教育審議会答申によると，チームとしての学校を実現するためには「専門性に基づくチーム体制の構築」や「関係者間の情報共有」等が重要である旨が明記されている。しかし，実際には養護教諭がけがの対応をする中で必要な情報が得られず，連携もうまくいかないケースがある。そこで，本研究では，けが対応事例の収集と分析を通して，けが対応時に養護教諭が必要としている情報は何か抽出することを目的とした。ここでは，特に教員と連携するために必要とする情報に注目した。その結果，けが発生時から事後措置までの一連の流れの中で養護教諭が必要とする情報が36項目にも上ることがわかった。また，けが発生時から処置については，児童生徒に関わる情報がすべてを占めているのに対し，事後は児童生徒，保護者，学校組織とさまざまな立場や組織に関わる情報であることが明確となった。

Sponge-associated bacteria can produce bioactive compounds similar to the host. Here, the investigation of antioxidant properties of the yellow-red pigment produced by sponge-associated bacterium AGS112 was conducted using in vitro and in vivo analysis. The 16S rRNA gene sequence showed the sponge-associated bacterium AGS112 has the closest similarity with Bacillus haikouensis C-89 (99%). The crude pigment extract produced by Bacillus haikouensis AGS112 contained both phenolic and flavonoid. The peaks occurred at 412 nm and 664 nm, which indicated as the presence of carotenoids and flavonoids, respectively. The active fraction extract obtained through bio-autography TLC, and had more potent antioxidant activity against DPPH (2,2-diphenyl-1-picrylhydrazyl) compared to the crude pigment extract with an IC50 value of 68.62±0.59 µg/ml and 198.88±1.66 µg/ml, respectively. Moreover, the active fraction extract at the concentration of 35 µg/ml could better enhance the viability of Schizosaccharomyces pombe and achieve the highest expression of sod1 and ctt1 genes. Metabolite profiling using LC-MS analysis confirmed the active fraction extract contains eudesmin and artelastin that might be contributed as antioxidants. These results suggest that the active fraction extract of the yellow-red pigment produced by B. haikouensis AGS112 had potency to be used as candidate for natural antioxidants.

type:text [要約] 中学校の理科の授業では年間1,000件以上の負傷・疾病が報告されており，理科の観察・実験活動の安全な実施には危機管理の観点は必要不可欠なものである。万一，ケガを伴う事故が理科の授業中に起きた場合，当該生徒と理科担当教員だけでなく，養護教諭，管理職，保護者との連携が必要となる。この連携の第一歩となるのが，理科教員と養護教諭の連携である。ところが，両者の養成カリキュラムには教職を意識した学習・交流の機会は皆無であり，このため，事故発生時の対応・連携について何の知識もないまま，教員になっているのが現状である。そこで，この問題の解決を目指して理科教員と養護教諭の志望学生が共に学ぶ合同のロールプレイ演習教材を開発した。本報告ではロールプレイ演習教材開発の経緯について述べる。

type:text [要約] 学校における危機管理には，「事前」，「事故等発生時」，「事後」の３つの段階があり，学校危機管理においては，事故の未然防止だけではなく，事故発生を想定したクライシスマネジメントも重要である。事故対応をシミュレートすることは，いざ事故が起きたときの冷静な対応につながる。学校事故に関する様々な事態の想定は教員として不可欠なものである。そして，新任として教職に就き，ひとたび事故が発生すれば当事者としての対応が求められる。そこで，理科教員と養護教諭の志望学生が共に学ぶ合同のロールプレイ演習教材を開発した。本報告では開発したロールプレイ演習教材を用いた授業実践について報告する。授業実践に参加した学生の学びについて質問紙により調査したところ，開発した教材を用いた授業は，お互いの職と連携を意識することのできる良い機会をつくり出すことが出来ていた。さらに，参加学生が事故発生時の対応の流れを多様な視点から確認できたことや，理科および養護教諭の教員志望学生双方の連携意識が強まったことも推察された。

理科の実験に活動おいて安全な活動は最も重要なものであり， 事故や負傷の未然防止は欠かせない. 多くの場合， 授業は教員の裁量で進行するものであり， 安全に関する指導や配慮についても教員の知識や経験に委ねられている. そこで本研究では， 授業中の事故やケガ防止のための安全意識について理科教員に調査した. 調査対象は公立中学校に勤務する理科担当教員とし， 例として挙げる実験の題材は中学校1年生で行われることの多い「白い粉の実験」とした. 調査では， 白い粉の実験における安全配慮事項について自由記述形式で回答を得た. その回答から「火傷， 換気， 保護眼鏡， 食べる， アレルギー， 粉じん爆発」の各項目についての内容があったものを計上した. その結果， 8割を超える教員が火傷については意識的に注意していることがわかった.

主体的で創造的な力の養成のため2022年度より理数探究基礎， 理数探究の授業が開始された. しかし， 探究指導において課題設定の指導に教員は困難を感じている.本報告では一斉指導によりシステマティックに探究課題設定が可能となる授業法の開発を試みた. 今回， 探究課題設定で用いるブレインストーミングとその結果の整理方法として文殊カード法とマンダラチャートを組み合わせた指導法を試行した. さらにChatGPTが課題検討時のブレインストーミングの支援手段として活用可能かについて検討した. この結果， 文殊カード法によるブレインストーミングとその成果を， マンダラチャートを用いて整理する方法は取り組みやすいことが示唆された. また， ChatGPTが生成する課題は， 受講生にとっても興味があるものであり， ブレインストーミングにより抽出されたものと同等に扱えることが示唆された.

What is the status of lesson study in Vietnam and Japan? Is there a significant difference between the teaching experience of secondary school teachers and how they perceive the relevance of lesson study to their professional development in Vietnam and Japan? The mentioned questions were explored by collecting data from high school teachers (n = 197) in Vietnam and Japan through questionnaires. Descriptive statistics are used to analyze the data. The results show that the involvement of secondary school teachers in lesson study and the contribution related to their professional development is very high. It can also be seen that there are significant differences between the teaching experience of secondary school teachers and how they perceive the relevance of lesson study to their professional development. The findings suggest that relevant education policies and appropriate funding need to be implemented to ensure that professional learning communities remain vibrant in both Vietnam and Japan. This is the key to enhancing secondary teacher competence, which can lead to higher student learning outcomes.

コロナ禍での新たなグローバルな科学教育方法としてオンラインでのSDGsワークショップをアジア・アセアン教育研究センターで開発した．このオンラインプログラムはこれまで千葉大学で実施してきた対面でのSDGsワークショップを基盤とし，オンラインの強みを生かし，アジア全域の高校生の参加も可能にするものとして開発した．2021年8月に連携大学との協働実施により日本，タイ，インドネシア，ベトナム，台湾の高校生60名が参加し，開催された．実施後のアンケートの自由記述のテキストマイニング解析により，非常にプログラム内容に満足するとともに，運営に当たっては，ネット環境および機材の性能に依存した通信トラブル，さらに会議，学習システムへの習熟などの課題が見いだされた．今後，ポストコロナに対応したより多くの国からの参加が可能な方式を目指す．

理科の教育目標は，観察，実験を通した学習を前提としている.これには，理科の観察，実験は安全に行われることが大前提である.しかし，理科の授業中に起こる負傷・疾病は計上させているものだけでも年間1，000件を超える.理科の授業中に事故が発生してしまった場合，第一発見者となるのは理科教員である.このため，事前のケガをしてしまった時のことの想定と，養護教諭との連携が欠かせない.本研究では理科教員と養護教諭を対象として，理科における危機管理の実態を明らかにするために質問紙調査を行った.本報告では，理科教員と養護教諭の校内体制の実状況と事故発生時の意識の差について報告する.調査の結果， 理科教員， 養護教諭共に校内の緊急・救急体制における教職員の意識はおおむね十分であると感じているものの，理科の授業中の事故発生を想定した場合の理科教員の自身の役割に関する意識は低い可能性があるという実態が示唆された.

Fruit peels, pericarps, or rinds are rich in phenolic/polyphenolic compounds with antioxidant properties and potentially beneficial effects against obesity and obesity-related non-communicable diseases. This study investigated the anti-obesity effects of matoa (Pometia pinnata) and salak (Salacca zalacca) fruit peel. Neither matoa peel powder (MPP) nor salak peel powder (SPP) affected the body weight, visceral fat weight, or serum glucose or lipid levels of Sprague–Dawley rats when included as 1% (w/w) of a high-fat diet (HFD). However, MPP significantly decreased the hepatic lipid level. MPP at a dose of 3% (w/w) of the HFD decreased body weight, visceral fat, and serum triglyceride levels as well as the hepatic lipid content. The inhibitory effect of MPP on hepatic lipid accumulation was not enhanced when its concentration was increased from 1% to 3% of the HFD. The anti-obesity effect of matoa was partly explained by the inhibitory effect of the matoa peel extract on fatty acid-induced secretion of ApoB-48 protein, a marker of intestinal chylomicrons, in differentiated Caco-2 cell monolayers. We identified hederagenin saponins that are abundant in MPP as potential anti-obesity substances. These results will contribute towards the development of functional foods with anti-obesity effects using the matoa fruit peel.

<p>千葉大学が実施している国際的な教師インターンシップであるTWINCLEプログラムでは， 千葉大学教育学部を中心とした学生が東南アジアの学校で理科の実験を含む授業を英語で実践している. 我々は振動と音の関係を説明する授業を開発し，2019年3月にベトナム・ハノイの中学校と高校で実施した. この授業には，STEM教育における科学教育の目標に沿って，社会における技術との関連を持たせた. さらに，教材として紙コップのスピーカを採用し，授業中に生徒が自分で組み立てられるように改善した. 授業後，生徒による授業評価アンケートを実施し，生徒の回答を分析した. この結果，ほとんどの生徒が授業は楽しく，知識が深まったと感じていた. 一方，英語での説明がよくわからないという回答があった. 言語の壁があったとしても，視覚教材と実験を組み合わせることで生徒にとって有意義な授業の提供が可能であることが示唆された.</p>

<p>千葉大学では平成26年度より文部科学省大学教育改革加速プログラム（AP）タイプIII（高大接続）「次世代才能スキップアップ」プログラムを千葉県内及び近隣都県高等学校と連係し実施してきた．この中で高校生を対象とした国際研究発表会を実施してきた．これは高校生が研究発表を海外の研究者・留学生に対して英語で行うものである．今回，平成28年度から平成30年度に参加した高校生の気づきと学びについて，アンケートの記述をテキストマイニング法により解析した．この結果，生徒は英語での会話に関して難しさを感じるとともに，研究発表は成功したと感じていることが示された．本プログラムは生徒に英語コミュニケーションの重要性に気付かせ，英語発表の自信につながると考えられる．</p>

<p>Chiba University has carried out a student exchange program called "Twin college envoys (TWINCLE)" as part of Japan's re-inventory project with the support of MEXT since 2012. The TWINCLE program has 12 partner universities in 5 ASEAN countries—Indonesia, Thailand, Singapore, Cambodia, and Vietnam. During the 5 years of the program, 246 ASEAN students visited Japan. This study analyzes the effect of the program, focused on the influence on the careers of ASEAN students through the program activities, using students' comments in activity reports from 2013 to 2016 using text-mining methods and subsequently cluster analysis methods. The results suggest that ASEAN students were interested in each activity in the TWINCLE program and had good impressions after participating in the program.</p>

千葉大学では平成 24 年度より文部科学省大学世界展開力強化事業「ツインクル」プログラムをASEAN 諸国 12 大学と連係し実施してきた．今回，平成 25 年度平成 28 年度に参加した ASEAN 諸国の留学生の気づきと学びについて，アンケートの記述をテキストマイニングの手法により解析した．この結 果，カテゴリとしてプログラム内容に関する単語とともに culture を中心として科学や，学校に関するさまざまな Japan/Japanese に関する学びがあることを示す結果が得られた．さらに学生が日本での大学院での学びや日本での職業選択への希望が見出された．したがって本プログラム内容が，留学生の親日的 感情を高め，キャリア意識の形成への影響が示唆された．

&lt;p&gt;The purpose of this study was to investigate how findings in science education findings play a key role in the improvement of science lessons for secondary school students. Japanese university students created a science lesson on tomography for Singapore students as part of the TWINCLE program. We investigated the change of lesson plans, PowerPoint slides, teaching materials and worksheets on the topic of tomographic visualization before and after the lesson with our Singapore counterparts. The results of this study show three key points: 1. The process of improvement of the science lesson became a form of active learning for the university students. 2. The university students&#039; lesson plans and teaching materials were revised according to the context so as to find the location of blood clots present in blood vessels. 3. The revised hands-on activity exposed secondary school students to a more &quot;authentic&quot; application of the concept of tomography.&lt;/p&gt;

<p>The TWINCLE program is a teacher internship program which aims to enhance mutual understanding of foreign cultures and to develop students' ability to succeed in overseas internships. In this study, reports written by students who participated in the program were analyzed by a text mining method to evaluate the changes in their ideas about the development and practice of the teaching materials. The results indicate that the students' attitudes towards science classes changed from 'simple lecture oriented classes' to 'problem-solving learning oriented classes'. Therefore, participating in the TWINCLE program is an effective way to change the students' image of the science class.</p>

Sponge associated bacteria with diverse NRPS-PKS genes could be the alternative source of bioactive compounds. This study aimed to analyze the activity as well as the regulatory mechanism regarding the anticancer properties derived from these bacteria against cancer cell lines. Extraction of bacterial metabolites using ethyl acetate solvent resulted yield ranging from 0.003 to 0.016% (w/v). Six extracts of bacterial metabolites showed markedly cytotoxic activity against 8 human hematopoietic cancer cell lines, with IC50 value 27.24 to 533.73 µg mL−1 in a dose dependent response, as assessed by MTT test. Supporting those results, reduction of cell density, morphological changes including cell shrinkage and formation of the apoptotic body of cancer cells lines, were found. As it might be due to the inhibitory effect of bacterial extract treatment. Apoptosis assay determined by annexin V-FITC apoptosis analysis showed that apoptotic cells were induced by bacterial extract after 6 h of treatment. Thus suggesting that the extract could be one of the external signals that activate the signal transduction pathway of apoptosis, especially in MOLT4 cells, acute lymphoblastic leukemic cell lines. Cell cycle analysis using propidium iodide revealed that the extract HAA 01 caused cell cycle arrest, since most of the MOLT4 cells were accumulated in Sub-G1 phase after 24 h of extract treatment. Interestingly, glycerol, lactose and amylum were the more preferable carbon source to produce anticancer compounds, than glucose. Our study indicates that anticancer activity of HAA 01 (identified as Bacillus subtilis based on API identification) extract is highly correlated with NRPS-PKS gene expression which is likely regulated by the type of carbon source.

© 2015, International Journal of Pharmacy and Pharmaceutical Science. All rights reserved. Objective: The objectives of this study were to isolate bioactive compounds from sponge-associated bacteria and analyze their cytotoxicity against MOLT4 leukemic cell lines through apoptosis. Methods: Crude extract of sponge-associated bacteria encoded SAB E 35, SAB E 38 and SAB E 40 was obtained by extraction using ethyl acetate. The cytotoxicity assay was determined by Cell Counting Kit 8. Apoptosis assay was determined by Annexin V Apoptosis Detection Kit. Further, cell cycle was determined by DNA-specific dye Propidium Iodide(PI). Results: The ethyl acetate extract showed cytotoxicity effect on MOLT4 cell line. Based on cytotoxicity rate value, SAB E 35 has the strongest capability of inhibiting MOLT4. The estimated ethyl acetate extract concentration required to inhibit the growth of MOLT4 cells by 50% (IC50) was 263.93 μg/ml. SAB E 35 showed the strongest apoptosis effect on MOLT4 cell line based on its ability to decrease viable cells, that remained22.70% and 37.07% cells on their late apoptosis phase. Cell cycle on treated cells resulted decrease in the number of cells in S phase, G0/G1 and G2/M, in addition to the accumulation of cells in sub-G1 fraction. Conclusion: The results of this study demonstrated that sponge-associated bacterial extracts have cytotoxicity effect on MOLT4 cell lines. Other results indicated that apoptosis occurred in MOLT4 treated by the extract, confirmed by the result of cell cycle analysis. The results suggested that the apoptotic effects of sponge-associated bacteria on MOLT4 can provide contribution to their claimed-anticancer activity.

セイヨウオトギリソウは、発熱性疾患の解熱、利尿促進、創傷治癒など様々な疾患に効く薬草として、古くから使用されている。最近、セイヨウオトギリソウは軽度のうつ症状を改善する薬草として着目され、うつ症状改善の分子機構の解析も精力的に行われている。一方、創傷治癒促進効果の機構については不明な点が多く、また分子機序を調べた研究が非常に少ない。本研究では、セイヨウオトギリソウ成分をエタノールで抽出し、ヒト線維芽細胞を用いたin vitro 創傷治癒モデル系における影響を調査した。セイヨウオトギリソウ抽出物は、2%ウシ胎児血清を含む培養条件では、4 μg/mL以下の低濃度においてヒト線維芽細胞の創傷部位とみなされる隙間空間への移動を促進した。6-8 μg/mLでは濃度依存的に細胞の移動促進効果が失われ、10 μg/mLでは、細胞の移動を阻害した。1%ウシ胎児血清を含む培養条件では、セイヨウオトギリソウの阻害効果の方が強く、移動促進のピークは観察されなかった。また、線維芽細胞の創傷部位への移動に関与するフィブリン溶解活性、およびウロキナーゼ型プラスミノーゲン活性化因子およびプラスミノーゲン活性化因子阻害因子-1の発現は、4-6 μg/mLのセイヨウオトギリソウ抽出物添加により促進されたが、10 μg/mLでは促進効果が消失した。以上の結果より、セイヨウオトギリソウ抽出物には、in vitro モデル系における創傷治癒を促進する成分と阻害する成分の両者を含むことが示唆された。

Shinrin-yoku, forest-air bathing and walking, is expected to be beneficial for the health of humans, but its effectiveness has not been clarified unequivocally by biophysical and biochemical analysis. In the present study, the amounts of oxidative damages after forest walking were compared with those after walking in an urban environment. Twelve female patients with rheumatoid arthritis (RA) aged 48-62 years-old and eleven healthy female volunteers aged 48-52 years-old walked in Chiba City at noon for 1 hr, and then moved by bus to a forest park on the first experimental day, followed by staying for one night in the park area. On the next day, they walked in the forest at noon for 1 hr according to their physical ability. RA patients in one group after forest walking showed decreased amounts of serum hydroperoxide and increased urine 7,8-dihydro-8-oxodeoxyguanosine (8-OHdG) levels, as estimated by a Diacron-Reactive Oxygen Metabolites (dROM) assay and an ELISA method, respectively, in comparison with those after city walking. In RA patients, the amounts of salivary IgA (sIgA), as estimated by an ELISA method, increased after forest walking. The amounts of serum matrix metalloproteinase-3 (MMP-3), as estimated by an ELISA method, decreased after forest walking.These results were concordant with the notion for the suppression of oxidative stress and modulation of immunological and/or enzymatic physiological functions by forest walking.

Objective: The aim of the present study was to investigate the role of DYX1C1 gene mutations as a candidate gene for developmental dyslexia (DD) in Japanese children. Cases and Methods: 15 Cases of DD, 22 non-DD family members of the dyslexia cases, and 60 normal volunteers, participated in this study. We performed screening tests for DYX 1C1 genes (exon 2 -3G &gt A, exon 10 1249G &gt T) by PCR mismatch assay. In addition to these 2 single nucleotide polymorphism (SNP) screened, we examined another SNP of exon 10 1259C &gt G, by DNA sequences and analyzed the association between these 3 SNPs and DD. Results: This study did not support the involvement of DYX1C1 gene variants in this sample of DD and their family members. Conclusion: We were unable to replicate the supportive findings between the DYX1C1 gene and DD in Japan, perhaps due to genetic and phenotypic heterogeneity. © 2011 Japan International Cultural Exchange Foundation &amp Japan Health Sciences University.

We hypothesised that the oxygen supply to the fatigued muscles is improved after the recovery with exercise caused by aerobic metabolism in the slow twitch fibres during the recovery period. Ten males performed a 30s maximum cycling (1st Exercise), followed by a 20min rest interval (Interval Rest) in which participants were either sitting (No Exercise) or low intensive cycling (Active). Then they again underwent a 30s bout of maximum cycling (2nd Exercise). The total work of the 2nd Exercise was higher in Active compared to No Exercise (297 +/- 14 vs 276 +/- 23 J center dot kg-1, P0.01). After Interval Rest, the muscle oxygenation level (P0.05) and blood lactate concentration (P0.05) were lower in Active compared to No Exercise. In Active, the total work was higher in the 2nd Exercise than the 1st Exercise (297 +/- 14 vs 277 +/- 23 J center dot kg-1, P0.01), and muscle oxygenation levels during the 2nd Exercise were also higher at 10 (P0.05) and 15 (P0.01) s after the beginning of the exercise. It was suggested that active recovery exercise would manage to increase the muscle oxygenation level, and improve the performance during the 2nd Exercise accompanied with blood lactate control.

日本における遺伝子変異を背景とする発達性読み書き障害(以下、ディスレクシアと記す)の有無を明らかにするため、DYX1C1遺伝子変異スクリーニングを実施した。多数の症例を検討するため、簡便かつ精度、経済性の優れたミスマッチPCR法を用いた。ディスレクシアの臨床症状のない正常群139名、臨床症状をもつ患者群12名と、臨床症状のない患者群の家族23名を対象とし、スクリーニングを行った。正常群からランダム抽出した20サンプル、患者、患者家族群の全サンプルはダイレクトシークエンスでその遺伝子型を調べ、スクリーニングシステムの精度を確かめた。患者、患者家族群は正常群よりも高頻度でDYX1C1遺伝子のエクソン2Elk-1転写因子結合部位変異(-3G → A)を示した。

The effects of active recovery using previously active and inactive muscle groups on power output and respiratory responses were examined. Ten male volunteers underwent two exhaustive 40-s bouts of leg cycling (1st Ex and 2nd Ex), separated by a 20-min recovery period. The recovery conditions were leg (Leg-Active) or arm (Arm-Active) cranking at 50% ventilatory threshold (VT), or sedentary control (Passive). The total output work (Total work) during the 2nd Ex in the Leg-Active condition was significantly higher than that in the Passive (299 vs. 282 J . kg body mass(-1) (J . BM(-1))). The values of Total work, peak (V) over barO(2) and peak heart rate during the 2nd Ex were significantly higher than those during the 1st Ex in both Leg- and Arm-Active. Total CO(2)excess after the 1st Ex was significantly higher than that after the 2nd Ex (67.6 vs. 26.0 ml . BM(-1)) in Passive. After the 2nd Ex, the Total CO(2)excess in the Leg-Active (51.5 ml . BM(-1)) was significantly higher than that in both the Passive (26.0) and Arm-Active (36.5), with Arm-Active being significantly higher than Passive. The recovery exercise using previously inactive muscles improved respiratory compensation ability related to performance enhancement.

Delayed healing of skin wounds can be caused by wound instability, whereas appropriate massage or exercise prevents sclerosis and scar contracture. However, the mechanism by which wound healing is related to mechanical stress has not been fully elucidated. The present study aimed to identify whether mechanical stretching of fibroblasts reduces their production of extracellular matrix. We transferred skin fibroblasts into collagen-coated elastic silicone chambers, cultured them on a stretching apparatus, and used RT-PCR to examine the effects of mechanical stretching on the expression levels of 17 genes related to extracellular matrix production and growth factor secretion. We found that connective tissue growth factor (CTGF) was downregulated after 24 hr of cell stretching. Specifically, the CTGF mRNA and protein levels were 50% and 48% of the control levels, respectively. These findings suggest that cyclic stretching of fibroblasts contributes to anti-fibrotic processes by reducing CTGF production.

ディスレクシア(dyslexia)とは、知能の遅れや注意・意欲の欠如などがないにも関わらず、文字や数字の読み書きに著しい困難を示す状態のことである。家族例もあり、すでにDyslexia の発症に関連する候補遺伝子が現在までに4 個報告されている。その1つとしてDYX1C1 遺伝子の変異とディスレクシアの発現との関連がフィンランドのTaipale らのグループにより報告された。ディスレクシアの早期発見のスクリーニング手法としてDYX1C1 遺伝子のエクソン2 のElk-1 転写因子結合部位変異(-3G→A)およびエクソン10 のコドン変異(1249G→T)に着目し、peptide nucleic acid (PNA)クランピングによる塩基多型スクリーニングシステムの開発を試みた。ディスレクシアは、通常の学習方法では文字の習得が困難であり、この学習の失敗経験のため自己評価の低下、不登校などの二次障害が起こることが示唆されている。二次障害を防ぐため、本人の自己評価が低下する以前に発見し、適切な教育および支援を行うことが望まれる。

今回、我々は7名の被験者から得たパラボリックフライト(PF)前後の血清を用いて、ヒトRSb細胞のUVに対する変異調節活性の変動について調査した。UV照射後の変異誘導はPCR-based differential dot-blot hybridization法とPeptide nucleic acid(PNA)クランピング法によるKーrasコドン12の塩基置換変異検定、そして、ウワバイン抵抗性の出現頻度によって解析を行った。PF前後の血清を細胞培養液中に1/1000量添加し、細胞を処理した結果、被験者の一人で、PF後の血清に変異誘導作用が認められた。他の6名の被験者の血清はPF前後とも変異誘導作用は見られなかった。しかし、それらの血清成分をBlue Bカラムクロマトグラフィーにより分離分画したところ、得られた8画分で変異誘導活性が検出された。以上の結果から、ヒト血清は変異調節作用因子を含んでおり、その活性はPFにより変化するということが示唆された。

One of the most intriguing biological subjects is cell-surface molecules that regulate the susceptibility of human cells to cell-killing effects after irradiation with far-ultraviolet light (UV, principally 254 nm wavelength). Human RSa cells have unusual sensitivity to UV-induced cell-killing. We searched for molecules on the cell-surface of RSa cells that were present in different amounts as compared to a variant of these cells, UVr-1 cells, which have increased resistance to UV cell-killing. Among the 21 molecules examined, the amount of transferrin receptor (TfR) protein was found to be 2-fold higher in UVr-1 cells compared with in RSa cells. The amounts of this protein were also higher in the UV-resistant hematopoietic cell lines, CEM6 and Daudi, as compared to the UV-sensitive cell lines, Molt4 and 697. Culturing of UVr-1 cells in a medium containing anti-transferrin antibodies resulted in sensitization of the cells to UV cell-killing as demonstrated by colony formation assay. Similar results were observed by treatment of the cells with TfR siRNA. In contrast, overexpression of TfR protein led to a resistance to UV cell-killing in RSa cells and monkey COS7 cells as demonstrated by both colony formation and apoptosis assay. In TfR-overexpressing cells, reduction of p53 and Bax protein was observed after UV-irradiation. Thus, TfR expression appears to be involved in the regulation of UV-resistance, possibly via modulation of the amount of p53 and Bax protein.

Culture of human cells with human interferon alpha and beta (IFNA and IFNB) results in increased resistance of the cells to cell killing by X rays. To identify candidate genes responsible for the IFN-induced X-ray resistance, we searched for genes whose expression levels are increased in human RSa cells treated with IFNA, using an mRNA differential display method and Northern blotting analysis. RSa cells, which showed increased survival (assayed by colony formation) after X irradiation when they were treated with IFNA prior to irradiation, showed increased expression levels of LEU13 (IFITM1) mRNA after IFNA treatment alone. In contrast, IF, and F-IFr cells, both of which are derived from RSa cells, showed increased X-ray resistance and high constitutive LEU13 mRNA expression levels compared to the parental RSa cells. Furthermore, the IFNA-induced resistance of RSa cells to killing by X rays was suppressed by antisense oligo-nucleotides for LEU13 mRNA. LEU13, a leukocyte surface protein, was previously reported to mediate the actions of IFN such as inhibition of cell proliferation. The present results suggest a novel role of LEU13 different from that in the inhibition of cell proliferation, involved in IFNA-induced refractoriness of RSa cells to X rays. (C) 2003 by Radiation Research Society.

Under the 1 G condition, the increase in antipain-sensitive protease activity promptly after UV (mainly 254 nm wavelength) irradiation in cultured human cells is detected and found to be one of the intriguing events involved in suppression of cell mutability. It was found that two cell lines. RSa and its variant UVAP-1 cells are applicable: the former is hypermutable and not susceptible to protease activation. while the latter is hypomutable and susceptible. In the present study it was investigated whether the increase in protease activity by UV irradiation is also observed in hypomutable human UVAP-1 cells exposed to gravity-changing stress and whether the increase is involved in suppression of UV mutagenicity. Exposure of human UVAP-1 cells to gravity-changing stress such as free-fall and parabolic flight prior to UV irradiation resulted in a pronounced increase in protease activity. but not to hypergravity conditions (2 and 10 G) prior to UV irradiation. To characterize the proteases, components of lysates from the cells exposed to free-fall prior to UV irradiation were fractionated by high performance liquid chromatography, indicating two separate fractions with highly increased levels of E-64-sensitive protease activity. In the cells treated with E-64 during their exposure to free-fall. K-vas codon 12 base substitution mutation was detected after UV irradiation, although the mutation was not detected after UV irradiation alone. Thus, the increase in E-64-sensitive protease activity may be involved in the suppression of UV mutagenicity in UVAP-1 cells exposed to free-fall. (C) 2003 Elsevier Science Ltd. All rights reserved.

The induction of apoptosis by microgravity and/or gravity-changing stress is considered to be one of the important causes of cell death, although the molecular mechanisms of the apoptotic event remain unclarified. In this study, we established a cell line, GSA-1, from ethyl methanesulfonate-treated human RSa cells. GSA-1 cells were highly susceptible to apoptosis after a free-fall; 24.4% of these cells underwent apoptosis after free-fall, compared with only 6% of the RSa cells. The apoptosis of GSA-1 cells was augmented by ultraviolet (UV, principally 254-nm wavelength) irradiation before free-fall to a greater extents than those in RSa cells. The molecular mechanisms of apoptosis included p53 and Bax proteins; the expression of nuclear p53 and cytoplasmic Bax in GSA-1 cells increased at 4 h after free-fall irrespective of irradiation. In addition, the rate of removal of cyclobutane pyrimidine dimer (CPD) in UV-irradiated GSA-1 cells was higher in cells exposed to free-fall than in those under the 1-G condition. Our results suggested that in GSA-1 cells, free-fall accelerates apoptosis, and that this process is associated with the accumulation of p53 and Bax, as well as CPD removal. Thus, GSA-1 cells should be useful for investigating the mechanism of cellular response, including the induction of apoptosis under gravity-changing stress.

Continuous variation in the direction of the gravity vector leads to various cellular responses including modulation of gene expression. Complementary DNA (cDNA) array analyses are available to observe the variation of gene expression under different conditions. In this study, expression levels of 588 representative genes were compared using the Atlas(TM) human cDNA expression array in human fibroblast cells with and without 3-dimensional (3-D) clinostat. Five upregulated and 8 downregulated genes were detected. Among these genes, upregulation of XRCC1, and downregulation of ERB-B2 and p21(Cip/Waf1) were confirmed by RT-PCR. These results suggested that the gene expression levels of XRCC1, ERB-B2 and p21(Cip1/Waf1) were modulated by vector-averaged microgravity induced by 3-D clinostat in human fibroblast cells. Our findings may be a basis for the biological study of 3-D culture systems. (C) 2002 Elsevier Science Ltd. All rights reserved.

Background, Recent studies have shown that gravity-changing stress modulates expression levels of cell surface molecules on human lymphocytes. However, previous in vitro microgravity studies have been performed with lymphocytes treated with mitogenic agents. Hypothesis: The aim of the study was to test if exposure of cells to gravity-changing stress alone alters the expression levels of cell surface molecules. Specifically, we examined whether the expression of activation markers is altered after exposure of lymphocytes to combinations of microgravity and hypergravity. Methods: We used free-fall In parabolic flight for human subjects and a drop-shaft to expose peripheral blood mononuclear cells (PBMC) to gravity-changing stress. After such exposure, PBMC were isolated, and expression levels of CD69, CD23 and CD38 were estimated using three-color flow cytometry, Results: Increased percentages of CD69-positive cells were observed with PBMC from 3 of 4 volunteers who undertook 10 parabolic flights. Exposure of blood to gravity-changing stress in the drop-shaft increased both ratios of CD69-positive cells and levels of CD69 expression on T and B cells. In contrast, the percentages of CD23-positive B cells was decreased. However, gravity-changing stress was not always followed by significant alteration in CD38 expression. Conclusions: Our findings suggest that CD69 and CD23 might be useful markers that are up- and down-regulated, respectively, after exposure of lymphocytes to gravity-changing stress.

It is an intriguing question whether gravity-changing stress modulates human cell mutability. To resolve this problem, it is necessary to determine the cellular events leading to modulation. We previously detected protease activation just after UV (UVC, principally 254 nm wavelength) irradiation followed by hypomutability in cultured human cells. We here investigated whether UV-activated protease activity is affected in human UVAP-1 cells exposed to gravity-changing stress prior to UV irradiation.

It is important to clarify the molecular mechanisms of physiological responses of the human body to changes in gravity. Previous reports demonstrated that gravity-changing stress increases the human urinary concentration of 8-hydroxy-2'-deoxyguanosine (8-OHdG). However, it has yet to be clarified whether repetitive parabolic flight modulates the urinary concentration of 8-OHdG after exposure to gravity-changing stress. In the present study, the effects of the number of previous experiences with parabolic flight on urinary excretion of 8-OHdG and concentration of serum ACTH were examined in 12 healthy volunteers.

The search for genes responsible for the sensitivity of human cells to cell-killing effects of UV is an important area of biological research. To identify candidate genes responsible for UV sensitization, levels of mRNA expression were compared between W-sensitive RSa cells and UV-resistant variant UVr-1 cells, using a differential display method and Northern blot analysis. Messenger RNA expression levels of human Ras homologue enriched in brain (Rheb) and/or a Rheb-like gene were up-regulated and slightly decreased in UV-irradiated RSa and UVr-1 cells, compared to in mock-irradiated cells, respectively. RSa and UVr-1 cells, both of which were treated with antisense oligonucleotides for Rheb RNA, exhibited an increased resistance to UV cell-killing. It remains unclear why UVr-1 cells are resistant to UV yet express Rheb mRNA at high levels, However, the results of antisense experiments together with the up-regulation in W-irradiated RSa cells, suggest that Rheb is involved in the UV sensitization of both cells to UV cell-killing. (C) 2000 Academic Press.

In this study, we analyzed levels of excretion of 8-OHdG into urine before and after exposure to gravitational changes including both microgravity and hypergravity by an enzyme-linked immunosorbent assay (ELISA).

We have reported that human interferon, one of cytokines present in serum, can confer hypomutability on various human cells. On the contrary, we have also reported that serum factors from cancer patients can enhance cell mutability. Therefore, it seems likely that cell mutability is changed by cytokine-like serum factors in our body. It is one of important space problems whether the mutability of human cells is regulated in response to microgravity and hypergravity (gravitational stress). However, there is little information about cell mutability during such stress. In this study, we investigated whether the mutability is changed by exposing cells to human serum factors after gravitational stress.