TALE-mediated plant genome visualization

Live imaging of the dynamics of nuclear organization provides the opportunity to uncover the mechanisms responsible for four-dimensional genome architecture. Here, we describe the use of fluorescent protein (FP) fusions of transcription activator-like effectors (TALEs) to visualize endogenous genomic sequences in Arabidopsis thaliana. The ability to engineer sequence-specific TALEs permits the investigation of precise genomic sequences. We could detect TALE-FP signals associated with centromeric, telomeric, and rDNA repeats and the signal distribution was consistent with that observed by fluorescent in situ hybridization. TALE-FPs are advantageous because they permit the observation of intact tissues. We used our TALE-FP method to investigate the nuclei of several multicellular plant tissues including roots, hypocotyls, leaves, and flowers. Because TALE-FPs permit live-cell imaging, we successfully observed the temporal dynamics of centromeres and telomeres in plant organs. Fusing TALEs to multimeric FPs enhanced the signal intensity when observing telomeres. We found that the mobility of telomeres was different in subnuclear regions. Transgenic plants stably expressing TALE-FPs will provide new insights into chromatin organization and dynamics in multicellular organisms

マチズクリ トシテノ ソウゴウガタ チイキ スポーツ クラブ ノ ヤクワリ : チイキ ト クラブ ノ トウギョ ニ モトメラレル バ ノ マネジメント

The Ministry of Education hammer out a new policy of sports promotion in 1995. The nineteen districts have granted a bounty to enhance sports activity and to support formation of sports club in the community by the educational authorities for four years. The nineteen districts are faced with three problems to operate each club when the bounty program is over. In the first，the most serious problem is to gather funds to operate the club in a number of the districts. In a few districts which are established the autonomous organization for operating the club by its members，sports programs are being carried on the community. In the second problem，each club has to keep places and facilities for sports activities and to train leaders and instructors of sports programs. In the final，the most districts that have granted the bounty have to establish and redesign new systems encouraging sports. The purpose of this study were to examine the strategies encouraging and operating sports club in the districts and to apply the Field Management to control the sports clubs and communities. In this study，data was acquired through interviews with administrative officials in sixteen districts that were granted a bounty. The major findings are summarized as follows: (1) Some clubs that depended on a local self-goveming body lacked the intentions of cooperating the club management of the members and were not clarified the strategies developing the club and promoting sports. (2) Some clubs establishing the new systems operating the club developed a new network in cooperation with the Amateur Sports Association in the districts，the sports club committee in the community，and so on. The clubs were functioning as the field to communicate interactively to these organizations. These clubs were possessed of the mastermind managing the club and taking care of sports promotion and community development continuously. These results indicated that club management and community development by sports promotion have to involve in various organizations that have to be functional to promote sports in the community

ソウゴウガタ チイキ スポーツ クラブ ノ イクセイ オ メグル ジュエキシャ フタン ノ モンダイ : カイヒ セッテイ ニオケル キンガク ノ イミ カイシャク

Sport is a spontaneous activity and a personal pleasure activity originally. Someone must not be provided with the sport which has such nature，and the individual who has value in the activity fundamentally should invest money，time，and energy for oneself. In spite of such a situation，the problems of beneficiary burden exist in the community-based sports club. The purposes of this study were to clarify the problems of beneficiary berden concerned with establishment and management of the community-besed sports club about that logical background，and to examine the meaning interpretation of the amont of money in a setup of the club dues. In this study，date was acquired through interviews with administrative officials or club managers in three districts. The major findings are summarized as follows:(1) Cost burden consciousness doesn't permeate a local resident，because there is a low value for sports services in a community. (2) To be important about the setup of the club dues was to give a feeling of th advantage which gave meaning to the value of the club individually and socially to the member. (3) In case the beneficiary burden system is introduced at the sports club，it is necessary to examine the right and duty which are given to the club member. These results indicated that club management and community development by sports promotion have to involve in various organizations that have to be functional to promote sports in the community

Liganded Thyroid Hormone Receptor Inhibits Phorbol 12-O-Tetradecanoate-13-Acetate-Induced Enhancer Activity via Firefly Luciferase cDNA

Thyroid hormone receptor (TR) belongs to the nuclear hormone receptor (NHR) superfamily and regulates the transcription of its target genes in a thyroid hormone (T3)-dependent manner. While the detail of transcriptional activation by T3 (positive regulation) has been clarified, the mechanism of T3-dependent repression (negative regulation) remains to be determined. In addition to naturally occurring negative regulations typically found for the thyrotropin β gene, T3-bound TR (T3/TR) is known to cause artificial negative regulation in reporter assays with cultured cells. For example, T3/TR inhibits the transcriptional activity of the reporter plasmids harboring AP-1 site derived from pUC/pBR322-related plasmid (pUC/AP-1). Artificial negative regulation has also been suggested in the reporter assay with firefly luciferase (FFL) gene. However, identification of the DNA sequence of the FFL gene using deletion analysis was not performed because negative regulation was evaluated by measuring the enzymatic activity of FFL protein. Thus, there remains the possibility that the inhibition by T3 is mediated via a DNA sequence other than FFL cDNA, for instance, pUC/AP-1 site in plasmid backbone. To investigate the function of FFL cDNA as a transcriptional regulatory sequence, we generated pBL-FFL-CAT5 by ligating FFL cDNA in the 5' upstream region to heterologous thymidine kinase promoter in pBL-CAT5, a chloramphenicol acetyl transferase (CAT)-based reporter gene, which lacks pUC/AP-1 site. In kidney-derived CV1 and choriocarcinoma-derived JEG3 cells, pBL-FFL-CAT5, but not pBL-CAT5, was strongly activated by a protein kinase C activator, phorbol 12-O-tetradecanoate-13-acetate (TPA). TPA-induced activity of pBL-FFL-CAT5 was negatively regulated by T3/TR. Mutation of nt. 626/640 in FFL cDNA attenuated the TPA-induced activation and concomitantly abolished the T3-dependent repression. Our data demonstrate that FFL cDNA sequence mediates the TPA-induced transcriptional activity, which is inhibited by T3/TR

Risk Perception of Health Risks Associated with Radiation Exposure among Residents of Okuma, Fukushima Prefecture

Ten years have passed since the Great East Japan Earthquake and the subsequent Fukushima Daiichi nuclear power plant accident on 11 March 2011. Okuma is a town hosting the Fukushima Daiichi nuclear power plant. The evacuation order for Okuma was partially lifted in April 2019. To clarify factors associated with risk perceptions of radiation among the residents of Okuma, we conducted a questionnaire survey in January 2021. Our results revealed that resident anxieties regarding the health effects of radiation exposure from living in Okuma were independently associated with positive PCL-Specific score, recognition of the consultation services with radiation experts in the municipal government of Okuma, and planned request for consultation service regarding radiation exposure by radiation experts, along with being female and living with a child. It is important for radiation experts to promote periodic communication of risks with individuals on a small scale to provide accurate information about the health effects of radiation and to provide maternal and child healthcare services and support regarding child-rearing and radiation exposure, to reduce concerns about radiation exposure and facilitate healthy living and wellbeing in Okuma

Occupational Difficulties of Disaster-Affected Local Government Employees in the Long-Term Recovery Phase after the Fukushima Nuclear Accident: A Cross-Sectional Study Using Modeling Analysis

Local government officials play a central role in post-disaster community reconstruction. However, few studies have eported on the actual difficulties during a complex disaster involving a nuclear accident. A self-rated questionnaire survey was administered to a total of 583 public employees in four municipalities around the Fukushima Daiichi Nuclear Power Station. The relationship between universal occupational factors and radiation disaster-specific factors on job satisfaction and intention to leave the job due to radiation anxiety was evaluated using structural equation modeling. The results showed that interpersonal problems (β = -0.246) and service years (β = ‐0.127) were related to job satisfaction, whereas radiation-specific factors were not related to job satisfaction, and only to the intention to leave work due to radiation anxiety. A sense of coherence was associated with job satisfaction (β = 0.373) and intention to leave work due to radiation anxiety (β= -0.182), and it served as a moderator of the universal occupational factors and the radiation disaster-specific factors. Therefore, it is suggested that outcomes could be improved through increased stress coping capacity by providing support for relationships and radiation risk communication to public employees during the disaster recovery period

A study on the development of technical skills in nursing education : through the skill evaluation using simulated patients

本研究の目的は，模擬患者を導入した看護技術試験の効果の検討であった． 研究実施にあたっては，Ｂ大学倫理審査委員会の承認を得た後，被験者と模擬患者に口頭と文書による説明を行い，同意を得た． 調査は，Ａ短期大学の看護学科２年次生３０名を対象に，筆者らが作成した７段階評価の２３項目の質問紙を用いて行われた．手順としては，まず２００４年９月に質問紙を用いた看護技術試験前の調査を行った．１０月に看護技術試験のために「吸引」についての事例およびチェックリストを配布し，デモンストレーションを実施した．１１月に学生が患者役となる形式の技術試験を行った．１２月に模擬患者を導入した技術試験を実施し，その後に９月と同様の質問紙の調査を行った． 肯定的表現に７点，否定的表現に１点を配した質問紙の各項目を看護技術試験前後で比較・検討した結果，技術試験前の高得点は，「緊張感がある－緊張感がない」，「効果的－効果的でない」であり，技術試験後は「よい－よくない」，「有用な－有用でない」および「継続してほしい－継続してほしくない」であった．技術試験前の低得点は，「易しい－難しい」，「明確な－混乱した」であり，技術試験後は「易しい－難しい」，「明確な－混乱した」であった．技術試験前後で得点が上昇したのは，「易しい－難しい」，「有益－有益でない」であり，得点が低下したのは，「やる気がでる－やる気がでない」，「達成感がある－達成感がない」であった．得点が変化しなかったのは，「刺激的な－刺激のない」および「実力を発揮できる－実力を発揮できない」であった． 学生の看護技術の習得レベルを評価し看護実践力を向上させるために，模擬患者を導入した看護技術試験は効果的な方法であることが示唆されたが，学生に過度の緊張を与えず，コミュニケーションスキルが身につくような配慮の工夫が必要であると思われた．The purpose of this study was to investigate the effectiveness of a test of technical nursing skills using simulated patients. Prior to the study, we obtained the permission of the ethics committee of University B, and explained the research to the subjects and simulated patients in order to obtain their informed consent. A survey was carried out using a questionnaire constructed by the researchers comprising ２３ items with a７‐point Likert scale. The subjects were ３０ senior nursing students at Junior College A. In September ２００４, we conducted an initial survey using the questionnaire, and in October gave the subjects a paper patient and the lists for checking their nursing techniques, and a suction demonstration. In November, we gave an ordinary skills test, and in December, using the same questionnaire as before, we conducted a second survey after a test of technical nursing skills using simulated patients. Pre - and post - test analysis of the questionnaire responses（the most positive answer being awarded７ points and the most negative answer being awarded１point）, showed that pre - test high scores were obtained on the “nervous- not nervous” and “effective-not effective” items ; post-test，“good test-not a good test”, “useful-not useful” and “like to continue-not like to continue”. The pre-test low scores were “easy-difficult” and “clear-unclear” ; and post-test，“easy-difficult” and “clear-unclear”. The increased scores between pre- and post-test were “easy-difficult”, “beneficial-not beneficial” and the decreased scores were “motivation-no motivation” and “sense of achievement-no sense of achievement”. The items which were unchanged were “stimulating-not stimulating”，“developed practical ability-did not develop practical ability”. These results show that the test using simulated patients was an effective method of developing students’ nursing techniques. However, teachers need to pay attention to developing students’ communication skills and not create excessive tension for them when they have skills tests

マウスリンパ組織のレプチンレセプター発現細胞の検出

Leptin, the product of the ob gene expressed in adipocytes, is shown to influence energy intake and expenditure, proliferation of CD^ T cells, neovascularization and intracellular triglycerides homeostasis in non-adipocytes. Leptin acts on target cells through receptor (OB-R). There are at least five different types of OB-R in mouse due to alternative splicing from db gene transcripts.OB-Ra〜OB-Rd share identical extracellular and transmembrane domains and JAK binding consensus sequence at cytoplasmic domain. Only OB-Rb has an additional STAT binding motif and is essential for most of leptin' s physiological functions through JAK-STAT pathway. OB-Ra is also reported to transduct weakly leptin's signal through JAK-phospholyration pathway. On this paper we examined which kinds of cell express OB-Ra or OB-Rb in lymphoid and fat tissues of the mouse. Both types of OB-R were detected in thymus, spleen and gastrolienal fat tissue by RTPCR method, then the constitutive cells were separated from dissected tissues and cultured in GIT medium with 10% heat-inactivated FBS. Primary cultured lymphocytes isolated from thymus or spleen expressed both OB-Ra and OB-Rb. On the other hand, in adhesive cells dispersed with enzymatic digestion and primary cultured OB-Rb was not detected, though OB-Ra detectable.Similarly, primary cultured adhesive cells of gastrolienal fat tissue expressed only OB-Ra. It is therefore most parsimonious to conclude that only lymphocytes express OB-Rb and response effectively to leptin in thymus