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INTRODUCTION Occupational therapy often involves handwriting acquisition practices that include the non-dominant hand when improvements in the dominant hand function are not possible because of trauma or stroke. This study explored whether character tracing and using a pegboard can effectively improve the handwriting of the non-dominant hand. METHODS A randomised controlled trial involving 60 healthy university students aged ≥18 years was conducted. Participants were randomly assigned to the writing group, peg group or control group. The character recognition rate was evaluated by computer software. Furthermore, character quality and writing speed were evaluated by humans using global legibility scales. Evaluations were performed before the intervention (baseline) and on days 5 and 10 of the intervention. Using the non-dominant hand, the writing group traced characters on paper with a ballpoint pen, and the peg group used a pegboard for 15 min/day for 10 days. RESULTS Compared with the peg and control groups, the writing group showed significant improvements in the character recognition rate and global legibility scale score. However, the global legibility scale score did not improve to the same level as that achieved with the dominant hand. None of the evaluation scores of the peg group showed significant improvements compared with those of the control group. There were no significant differences in improvements in the writing speed of the writing and peg groups compared with the control group. CONCLUSION Tracing characters can improve the handwriting ability of the non-dominant hand, but using a pegboard may be less effective. Future research is needed to examine how much practice is necessary to improve the handwriting ability of the non-dominant hand sufficiently.

An increasing number of universities in Japan have been using English as a medium of instruction (EMI) to teach academic subjects. Despite its prevalence, few studies have investigated the competencies required to teach EMI courses. This knowledge gap creates challenges when designing training or support programs for EMI instructors. To contribute to filling this knowledge gap, I conducted semi-structured interviews with instructors of EMI courses at a Japanese university. In these interviews, I explored the difficulties they experienced, competencies they believe EMI instructors should have, and types of support they want to deliver EMI courses effectively. The findings revealed that EMI instructors recognized English competency as a necessary skill but also felt that wider pedagogical and communicative skills are equally important to manage students from diverse linguistic, social, and academic backgrounds. The types of support they wished for were diverse, including multiple human resources and facilities on campus. 日本で益々多くの大学が英語による専門科目の授業（EMI）を実施している。その普及度にも関わらずEMIを効果的に実施するのに必要とされる能力についてはあまり調査が行われていない。この知識の差があることでEMI教員を対象とした研修や支援プログラムの構築が困難になっている。この知識の差を埋めることに貢献するために、本研究ではある日本の大学でEMI授業を実施する教員を対象に半構造化インタビューを行った。インタビューではEMI教員が経験した困難や、EMI教員に必要だと思われるコンピテンシー、そしてEMIの授業を効果的に行う上で受けたい支援について調査した。結果、EMI教員は英語力を不可欠なスキルと認識する一方、多様な言語・社会・文化的バックグラウンドを持つ学生に対応するためより広い意味での教育・コミュニケーション能力も必要と感じていることが分かった。希望する支援は多様でキャンパス内の様々な人的資源や施設を伴うものであった。

In the last few decades, the number of universities offering English medium instruction (EMI) in Japan has increased rapidly. Yet, studies on the EMI experiences of Japanese university students are limited in number. Few studies have examined the EMI experiences of students in medical school, who tend to have high English proficiency given the intense preparation required for the entrance exam. To fill this knowledge gap, I investigated medical students’ views on their experiences of elective medical and non-medical EMI courses that they were required to take. An analysis of 93 survey and 7 interview responses revealed that the majority of them found the level of their EMI classes to be reasonable, while approximately 30% found it to be difficult, especially in terms of speaking, listening, and vocabulary. Most of the students who experienced these problems did not do anything to solve them, revealing the presence of unmet support needs. ここ数十年間の日本のEMI（英語による専門科目の授業）の増加は著しい。一方、EMIの急速な普及により実践が研究を先回っている現状があり、日本人の大学生のEMI経験を深く調査した研究は少ない。例えば、入試への集中的な準備期間により比較的英語能力の高い医学部生のEMI経験を調査した研究は稀である。この知識の欠落を埋めることに貢献するため、本研究では93名分の質問紙調査の結果と7名分のインタビュー調査の結果を用いて、日本の医学部生のEMIの授業における経験を調査分析した。結果、回答者のうち大多数の学生が授業レベルを丁度よいと評価し、極めて困難であると評価した学生は３割程度であった。困難を感じた学生は特にスピーキング、リスニング、語彙の側面を困難であると評価している。困難を経験した学生の多くは問題を解決するために何もしなかったと回答しており、少数派ながら一定の学生の満たされていない学習支援ニーズの存在が明らかになった。

The adenylation domain of nonribosomal peptide synthetase (NRPS) is responsible for the selective substrate recognition and its activation (as an acyl-O-AMP intermediate) during ATP consumption. DhbE, a stand-alone adenylation domain, acts on an aromatic acid, 2,3-dihydroxybenzoic acid (DHB). This activation is the initial step of the synthesis of bacillibactin that is a high-affinity small-molecule iron chelator also termed siderophore. Subsequently, the activated DHB is transferred and attached covalently to a peptidyl carrier protein domain via a thioester bond. Adenylation domains belong to the superfamily of adenylate-forming enzymes including acetyl-CoA synthetase, acyl-CoA synthetase and firefly luciferase. We previously reported a novel N-acylation reaction for an acyl-CoA synthetase (AcsA) that originally catalyzes the formation of a thioester bond between an acid and CoA, yielding acyl-CoA. This novel reaction was also confirmed for acetyl-CoA synthetase and firefly luciferase, but not yet for an adenylation domain. Here, we for the first time demonstrated the synthesis of N-acyl-L-cysteine by a stand-alone adenylation domain, DhbE. When DHB and L-cysteine were used as substrates of DhbE, N-DHB-L-cysteine was formed. A Vmax value of 0.0156±0.0008 units mg−1 and Km values of 150±18.3 mM for L-cysteine and 0.0579±0.0260 mM for DHB were obtained in this novel reaction. Furthermore, DhbE synthesized N-benzoyl-L-cysteine when benzoic acid and L-cysteine were used as substrates. Through the N-acylation reaction of DhbE, we also succeeded in the synthesis of N-aromatic acyl compounds that have never previously been reported to be produced by this enzymatic method.

Peer-led online communities have shown a strong potential to foster resourceful environments for people living with depression without restrictions of time, space, and stigma. However, due to the scarcity of research, many aspects of depression online support groups remain inconclusive. To contribute to filing some of the knowledge gap, an exploratory study was conducted to explore how online communities designed for people with depression contribute to the everyday lives of depression patients. A sample of 980 posts was selected systematically from a selected online community and demographic and clinical information of the users who made those posts were recorded. The results of the quantitative and qualitative content analyses of those data indicate that the online community can service as a place to exchange various types of informational, emotional, and coaching support with users of diverse backgrounds.

To examine whether brain electrical responses to environmental stimuli were influenced by emotional contexts, event-related potentials (ERPs) elicited by nonstartle probe tones were recorded from 13 student volunteers while they were viewing emotionally positive, neutral, and negative slides of the International Affective Picture System. The auditory stimuli consisted of high-deviant (2000 Hz, p=.08), low-deviant (1050 Hz, p=.08), and standard (1000 Hz, p=.84) tones with a mean onset-to-onset interval of 600 ins. Participants were told to ignore the tones. High-deviant tones elicited a larger N1 (peaking around 100 ms) when participants were viewing negative slides than when viewing positive slides. The amplitude of the P2 elicited by standard tones (peaking around 170 ms) was smaller when participants were viewing positive slides than when viewing negative and neutral slides. The amplitude of the mismatch negativity (150-200 ms) tended to reduce during positive slide presentation, but this difference appeared to be due to reduction of the P2 elicited by standard tones. These findings suggest that visually induced emotional states have a sequential effect on auditory information processing, in that the influence of negative emotion appears at an earlier stage than that of positive emotion.

It has been reported that proton pump inhibitors are more effective than H2 receptor antagonists in patients with functional dyspepsia. Dyspeptic symptoms that respond to proton pump inhibitors are classified as acid-related dyspepsia. A new questionnaire for assessing gastroesophageal reflux disease (GERD), the Frequency Scale for Symptoms of GERD, covers the 12 most common symptoms of GERD patients. A quantitative assessment of the changes of reflux symptoms and acid-related dyspepsia was made in GERD patients receiving proton pump inhibitor therapy. Sixty-eight GERD patients receiving proton pump inhibitor therapy completed the questionnaire before and after treatment for 8 weeks. There is a significant positive correlation between reflux symptoms and acid-related dyspepsia before and after therapy (r = 0.569 and r = 0.569; both P's < 0.001) and acid-related dyspepsia in patients with both nonerosive and erosive GERD. We conclude that GERD patients suffer not only from reflux symptoms, but also from acid-related dyspepsia, and proton pump inhibitors improve both types of symptoms.

Phosphatidylcholine biosynthesis via the CDP-choline pathway is primarily regulated by CTP:phosphocholine cytidylyltransferase (CT) encoded by the Pcyt1a and Pcyt1b genes. Previously, we identified an Ets-1-binding site located at -49/-47 in the promoter of Pcyt1a as an important transcriptional element involved in basal CTalpha transcription (Sugimoto, H., Sugimoto, S., Tatei, K., Obinata, H., Bakovic, M., Izumi, T., and Vance, D. E. (2003) J. Biol. Chem. 278, 19716-19722). In this study, we determined whether or not there were other important elements and binding proteins for basal CTalpha transcription in the Pcyt1a promoter, and if other Ets family proteins bind to the Ets-1-binding site. The results indicate the formation of a ternary complex with Ets-1 binding at -49/-47 and Sp1 binding at -58/-54 of the Pcyt1a promoter that is important for activating CTalpha transcription. When nuclear extracts of COS-7 cells expressing various Ets family repressors were incubated with DNA probes, binding of Net to the probes was observed. Net dose-dependently depressed the promoter-luciferase activity by 98%, even when co-expressed with Ets-1. RNA interference targeting Net caused an increase of endogenous CTalpha mRNA. After synchronizing the cell cycle in NIH3T3 cells, CTalpha mRNA increased at the S-M phase corresponding to an increase of Ets-1 mRNA and a decrease of Net mRNA. These results indicated that Net is an important endogenous repressor for CTalpha transcription.

本研究では、五感をまんべんなく使うことができる環境学習プログラムの開発を行い、その児童への学習効果について調査した.児童を対象に、五感を満遍なく使う環境学習として『五感しらべ』を4パターンの方法に分けてグループごとに実施し、それぞれのグループの情報量・内容について考察した. また、児童一人一人に対し、『五感しらべ』の前後に五感認識現状を知るツールとして『五感の履歴書』を実施し、『五感しらべ』によって児童の五感認識がどのように変化するかを考察した. それらの考察から、児童にとって五感学習効果が高くなる五感調査手法を提案した.

Background: We previously reported that a new endoscopic classification of gastroesophageal reflux disease, the Los Angeles classification, showed considerable observer variation depending on the experience of the endoscopist. In the present study, we evaluated some modifications of the classification to determine whether we could decrease observer variation. Methods: Fifty endoscopic photographs, each showing four images of the squamo-columnar junction, were prospectively obtained from 50 consecutive patients with gastroesophageal reflux disease. Two groups of eight endoscopists divided by their endoscopic experience, group 1 (100–500 procedures) and group 2 (more than 500 procedures), assessed the photographs using classifications with the following modifications: (i) addition of grade O to describe healed mucosal breaks and setting grade B as more than 5 mm or 10 mm; or (ii) addition of grade O and setting grade D as 75–99% or 100% circumferential. Results: Changing the definition of grade B or grade D did not increase the kappa values for either group of observers. Conclusions: These modifications of the Los Angeles classification were unable to decrease observer variation.

In a previous study we purified a novel lysoPLD (lysophospholipase D) which converts LPC (lysophosphatidylcholine) into a bioactive phospholipid, LPA (lysophosphatidic acid), from the rat brain. In the present study, we identified the purified 42 and 35 kDa proteins as the heterotrimeric G protein subunits Gα(q) and Gβ(1) respectively. When FLAG-tagged Gα(q) or Gβ(1) was expressed in cells and purified, significant lysoPLD activity was observed in the microsomal fractions. Levels of the hydrolysed product choline increased over time, and the Mg(2+) dependency and substrate specificity of Gα(q) were similar to those of lysoPLD purified from the rat brain. Mutation of Gα(q) at amino acids Lys(52), Thr(186) or Asp(205), residues that are predicted to interact with nucleotide phosphates or catalytic Mg(2+), dramatically reduced lysoPLD activity. GTP does not compete with LPC for the lysoPLD activity, indicating that these substrate-binding sites are not identical. Whereas the enzyme activity of highly purified FLAG-tagged Gα(q) overexpressed in COS-7 cells was ~4 nmol/min per mg, the activity from Neuro2A cells was 137.4 nmol/min per mg. The calculated K(m) and V(max) values for lysoPAF (1-O-hexadecyl-sn-glycero-3-phosphocholine) obtained from Neuro2A cells were 21 μM and 0.16 μmol/min per mg respectively, similar to the enzyme purified from the rat brain. These results reveal a new function for Gα(q) and Gβ(1) as an enzyme with lysoPLD activity. Tag-purified Gα(11) also exhibited a high lysoPLD activity, but Gα(i) and Gα(s) did not. The lysoPLD activity of the Gα subunit is strictly dependent on its subfamily and might be important for cellular responses. However, treatment of Hepa-1 cells with Gα(q) and Gα(11) siRNAs (small interfering RNAs) did not change lysoPLD activity in the microsomal fraction. Clarification of the physiological relevance of lysoPLD activity of these proteins will need further studies.

In the present study, we investigated the influence of cytological stains in analyzing DNA extracted from cytological slides by comparative genomic hybridization (CGH). Multiple imprint cytological slides were prepared for fresh-frozen breast cancer tissue samples and the slides were stained by three staining methods for each sample. Under microscopic observation, cancer cells were selectively microdissected from the slides and forwarded to DNA extraction, whole genome amplification, and CGH analysis. CGH was successfully performed for all methylgreen-stained and May-Grunwald-Giemsa (MGG)-stained cytological smear slides, but for two Papanicolaou (PAP)-stained slides. The number of chromosomal imbalances detected were 5-10 in methylgreen-stained slides and 5-9 in MGG-stained slides. The chromosomal imbalances resemble each other between methylgreen-stained and MGG-stained slides. The present study indicates that the MGG stain is preferred to the PAP stain for the purpose of cytogenetical analysis by CGH for DNA extracted from cytological smear slides.

A lysophospholipase D (lysoPLD) was purified to apparent homogeneity from rat brain nuclear fractions using 1-[(14)C]palmitoyl-glycerophosphorylcholine as a substrate. The abundance of autotaxin (ATX), a secretory lysoPLD, was also estimated for each fraction. The nuclear fraction had relatively high levels of lysoPLD activity but weak immunoreactivity with an anti-ATX antibody. LysoPLD activity was further purified 5550-fold by sequential chromatography. The final preparation migrated as a single band with a molecular weight of 35,000. Anti-ATX antibodies did not cross-react with the purified enzyme. Moreover, enzyme activity was highest at pH 7.0-7.5 and requires Mg(2+). The Km and Vmax values for 1-palmitoyl-glycerophosphorylcholine were 176 microM and 0.3 micromol/min/mg, respectively. The purified enzyme hydrolyzed saturated forms of LPC more robustly than unsaturated forms. The enzyme could hydrolyze platelet-activating factor (PAF) to the same extent as 16:0-LPC, and showed a higher activity toward lysoPAF (1-O-hexadecyl-2-lyso-glycerophosphorylcholine). These results suggested that the lysoPLD purified from rat brain nuclear fractions in this work is a novel enzyme that hydrolyzes lysoPAF, PAF, and LPC to liberate choline.

The effects of postprandial water intake on the gastrointestinal tract have not been systematically investigated in humans.In 8 healthy volunteers, the gastric antral pressure was measured with a strain gauge transducer, while the esophageal and lower esophageal sphincter pressures were measured with an infused catheter with a Dent sleeve. The esophageal pH at 5 cm above the lower sphincter was measured with a microglass electrode. A standard test meal (560 kcal) was eaten and 500 ml water was ingested 1 h later. The plasma cholecystokinin level was assessed at 4-min intervals. As a control, the same study was done on another day with sham water intake.At 4 min after water intake, there was a significant decrease in gastric antral motility and a significant increase in the plasma cholecystokinin level. Water intake also significantly increased the occurrence of gastroesophageal reflux.The rapid increase in cholecystokinin after water intake may be initiated by a feedback mechanism related to inflow of fatty chyme into the duodenum that inhibits gastric antral activity.

Background and aim An early and accurate evaluation by a general practitioner is needed to screen out non-gastroesophageal reflux disease (GERD) patients. A recent questionnaire (QUEST) highlighted problems with specificity and complexity, so the aim of the present study was to design a simplified questionnaire. Methods When admitted to hospital to undergo an upper gastrointestinal endoscopy for suspected GERD, 333 patients completed a 50-item questionnaire requiring 'yes/no' answers to different combinations of questions relating to symptoms of upper gastrointestinal tract conditions (e.g. GERD, ulcers and functional dyspepsia) and psychosomatic symptoms. The endoscopic diagnosis was then correlated with the rate of positive answers to each question. Results Based on the analysis of the 50 items, the 8-10 questions most often answered affirmatively by each of the GERD and non-GERD groups were chosen for the simplified questionnaire. Three draft questionnaires were compiled. After calculating the sensitivity, specificity and accuracy in relation to the diagnosis of GERD and other conditions, it was found that questionnaire B (selection of persons answering 'yes' to at least one of questions 1-5 and exclusion of persons answering 'yes' to at least three of questions 7-10) had a high sensitivity, high specificity and low false positive rate. Conclusion A novel questionnaire was developed. It was designed to detect the symptoms of GERD while simultaneously excluding non-GERD patients. This simplified nine-item simplified questionnaire had a sensitivity of 79.8%, a specificity of 53.6% and an accuracy of 63.4%.

The aim of this study was to produce a simplified questionnaire for evaluation of the symptoms of gastroesophageal reflux disease (GERD).A total of 124 patients with an endoscopic diagnosis of GERD completed a 50-part questionnaire, requiring only "yes" or "no" answers, that covered various symptoms related to the upper gastrointestinal tract, as well as psychosomatic symptoms. The 12 questions to which patients most often answered "yes" were selected, and were assigned scores (never = 0; occasionally = 1; sometimes = 2; often = 3; and always = 4) to produce a frequency scale for symptoms of GERD (FSSG). Sensitivity, specificity, and accuracy of the FSSG questionnaire were evaluated in another group of patients with GERD and non-GERD. The usefulness of this questionnaire was evaluated in 26 other GERD patients who were treated with proton pump inhibitors for 8 weeks.When the cutoff score was set at 8 points, the FSSG showed a sensitivity of 62%, a specificity of 59%, and an accuracy of 60%, whereas a cutoff score of 10 points altered these values to 55%, 69%, and 63%. The score obtained using the questionnaire correlated well with the extent of endoscopic improvement in patients with mild or severe GERD.This new questionnaire is useful for the objective evaluation of symptoms in GERD patients.

Phosphatidylcholine biosynthesis via the CDP-choline pathway is primarily regulated by CTP:phosphocholine cytidylyltransferase (CT). Transcriptional enhancer factor-4 (TEF-4) enhances the transcription of CTalpha in COS-7 cells by interactions with the basal transcription machinery (Sugimoto, H., Bakovic, M., Yamashita, S., and Vance, D.E. (2001) J. Biol. Chem. 276,12338-12344). To identify the most important transcription factor involved in basal CTalpha transcription, we made CTalpha promoter-deletion and -mutated constructs linked to a luciferase reporter and transfected them into COS-7 cells. The results indicate that an important site regulating basal CTalpha transcription is -53/-47 (GACTTCC), which is a putative consensus-binding site of Ets transcription factors (GGAA) in the opposite orientation. Gel shift analyses indicated the existence of a binding protein for -53/-47 (GACTTCC) in nuclear extracts of COS-7 cells. When anti-Ets-1 antibody was incubated with the probe in gel shift analyses, the intensity of the binding protein was decreased. The binding of endogenous Ets-1 to the promoter probe was increased when TEF-4 was expressed; however, the amount of Ets-1 detected by immunoblotting was unchanged. When cells were transfected with Ets-1 cDNA, the luciferase activity of CTalpha promoter constructs was greatly enhanced. Co-transfection experiments with Ets-1 and TEF-4 showed enhanced expression of reporter constructs as well as CTalpha mRNA. These results suggest that Ets-1 is an important transcriptional activator of the CTalpha gene and that Ets-1 activity is enhanced by TEF-4.

Pyrroloquinoline quinone glucose dehydrogenase (PQQGDH) was overexpressed in Escherichia coli , and its impact on sugar-dependent respiration was investigated. Sugar-dependent respiration patterns under PQQGDH overexpression can be devided into two types. The first type involves d -glucose and d -mannose, which are utilized by the phosphotransferase system (PTS) and are also the substrates of PQQGDH. As a result of PQQGDH overexpression, the apparent K m value of sugar-dependent respiration shifted to higher concentration compared with E. coli parental cells. The second type included d -xylose and d -galactose, which are the substrates of PQQGDH, but not the PTS sugars. PQQGDH overexpressing cells showed much higher respiration than parental cells. These results suggested that PQQGDH overexpression may alter sugar utilization preferences in E. coli , suggesting further possible applications in metabolic engineering for carbon source utilization.