Introducing Objective Structured Clinical Examinations for Undergraduate Orthoptic Education

Aim: Introducing the objective structured clinical examination (OSCE) to improve orthoptic education and clinical competence including the clinical attitude and skills of orthoptic students.Methods: Subjects were thirty-one fourth grade orthoptic students from our department. Six basic examinations were included in stations for trial use of OSCE. The OSCE were performed and compared before and after the eight weeks clinical training. Ten items of clinical attitude and six examinations were evaluated respectively. Six experienced orthoptists evaluated students\u27 clinical attitude and examination techniques while they were performing simulated patient (SPs) examinations according to the evaluation sheets we designed.Results: After clinical training, both students\u27 attitudes toward SPs and examination techniques including correction of refractive errors improved significantly (p<0.05). However, the refractive correction score did not reach sixty. Attitudes toward SPs improved in eight areas out of ten. Safety procedures during examination and signaling the end of examinations did not attain a score of sixty.Conclusion: We were able to evaluate objectively the effect of clinical training on the clinical competence (attitude toward patients and examination techniques) of students by OSCE. Improvements in the evaluation sheets and crossed-check system are still necessary

MHC class II DQB diversity in the Japanese black bear, Ursus thibetanus japonicus

BackgroundThe major histocompatibility complex (MHC) genes are one of the most important genetic systems in the vertebrate immune response. The diversity of MHC genes may directly influence the survival of individuals against infectious disease. However, there has been no investigation of MHC diversity in the Asiatic black bear (Ursus thibetanus). Here, we analyzed 270-bp nucleotide sequences of the entire exon 2 region of the MHC DQB gene by using 188 samples from the Japanese black bear (Ursus thibetanus japonicus) from 12 local populations. ResultsAmong 185 of 188 samples, we identified 44 MHC variants that encoded 31 different amino acid sequences (allotypes) and one putative. The phylogenetic analysis suggests that MHC variants detected from the Japanese black bear are derived from the DQB locus. One of the 31 DQB allotypes, Urth-DQB*01, was found to be common to all local populations. Moreover, this allotype was shared between the black bear on the Asian continent and the Japanese black bear, suggesting that Urth-DQB*01 might have been maintained in the ancestral black bear population for at least 300,000 years. Our findings, from calculating the ratio of non-synonymous to synonymous substitutions, indicate that balancing selection has maintained genetic variation of peptide-binding residues at the DQB locus of the Japanese black bear. From examination of genotype frequencies among local populations, we observed a considerably lower level of observed heterozygosity than expected. ConclusionsThe low level of observed heterozygosity suggests that genetic drift reduced DQB diversity in the Japanese black bear due to a bottleneck event at the population or species level. The decline of DQB diversity might have been accelerated by the loss of rare variants that have been maintained by negative frequency-dependent selection. Nevertheless, DQB diversity of the black bear appears to be relatively high compared with some other endangered mammalian species. This result suggests that the Japanese black bears may also retain more potential resistance against pathogens than other endangered mammalian species. To prevent further decline of potential resistance against pathogens, a conservation policy for the Japanese black bear should be designed to maintain MHC rare variants in each local population

Versatile Psychophysiological Potencies of Essential Oils, when Seen as a Function of Behavioral Task Assigned to the Participants after Inhalation

To elucidate the psychophysiological effect of inhaling essential oils, in this paper, we sought to assess the following 12 essential oils: basil, bergamot, cardamom, cinnamon, juniper, lemon, orange, palmarosa, peppermint, sandalwood, spearmint, and ylang ylang. As these being target odors, we focused on the verbal (semantic) and non-verbal (skin temperature) endpoints of the stimuli. In our experimental design, we managed to assign different behavioral tasks to the participants. The Uchida-Kraepelin test was used as a mental arithmetic task and listening to environmental (natural) sounds as an auditory task. In the verbal study, for an example, we conducted the sensory test twice, once before and once after the task. As a measure of the perceived odor quality in participants after inhalation of a given aroma, we employed a sensory evaluation spectrum. It is a bar graph in which the mean of the difference in score between pre- and post-task inquiry (post minus pre) was plotted against the impression descriptors. Taking into account of the obtained skin temperature changes between pre- and post-task inhalations, the subtle nuances between verbal and non-verbal expressions seen as a function of the two behavioral tasks assigned to the participant suggested that essential oils may have versatile psychophysiological potencies by the nature

リン脂質過酸化物の生物有機化学的合成

Chemoenzymatic synthesis of 1-stearoyl-2-hydropeoxyacyl-sn-glycerophospholipids including phosphatidylcholine (PC-OOH), phosphatidic acid (PA-OOH), phosphatidylethanolamine (PE-OOH), phosphatidylglycerol (PG-OOH) and phosphatidylserine (PS-OOH). The hydroperoxy acyl moieties were prepared via hydroperoxidation of linoleic, dihomo-γ-linolenic and arachidonic acids by soybean, potate lipoxygenase or autoxidation. Their hydroperoxy group was protected as a dimethylperacetal before condensation with lysophosphatidylcholine. Optically active lysophosphatidylcholine was prepared via short pathway involving lipase-catalyzed direct enantioselective stearoylation of 2-O-benzylglycerol and choline phosphate synthesis. Peroxy fatty acids and lysophosphatidylcholine thus obtained were condensed using dicyclohexylcarbodiimide in chloroform. Removing the peracetal group in the product and purification by reverse-phase chromatography afforded the desired PC-OOH’s. PA-OOH, PG-OOH, PE-OOH and PS-OOH were obtained by phospholipase-D catalyzed transphosphatidylation from PC-OOH. As a reference compound for biological studies of hydroperoxy phopholipid, PC-OH's were also prepared in which hydroxy unsaturated fatty acyl group was linked to the sn-2 position of the glycerophospholipids.健康正常人の血液などの組織中にはリン脂質過酸化物が極微量で存在し，疾病や老化によってその濃度が顕著に上昇する事が知られている．その事が明らかにされた当初は，極めて複雑な混合物をなす生体脂質中に含まれる不安定な極微量の過酸化脂質を単離・構造決定する事は殆ど不可能と考えられていた．現在もなお，そのような脂質過酸化物を生体組織から純粋に取り出し，構造決定したという報告は無い．従ってそのような分子種の化学的・生理学的性質は不明であったが，脂肪酸過酸化物が毒性を示す事から，リン脂質過酸化物もおそらく毒性を示すだろうと考えられてきた．このような漠然とした推定を科学的に明らかにするためには，化学合成によらざるを得ない．我々はこの未知の合成に取りかかった．しかし，従来の化学的手法のみでは不可能である事も明らかであった．その中で予想された困難の一つは極めて不安定なヒドロペルオキシ基を不飽和脂肪酸のある特定の位置にどのように導入するかという問題と，ヒドロペルオキシ基に影響を与える事なく合成中間体をどのように化学変換するかであった．第一の問題に対する解決策として，不飽和脂肪酸に大豆リポキシゲナーゼを作用させる事で解決する事ができた．植物に広く分布する酵素であるリポキシゲナーゼは植物中でリノール酸に作用して過酸化し，その生成物にヒドロペルオキシドリアーゼという酵素が作用して種々のアルデヒドが精製し，これは植物の青臭みを与える．第二の問題に対しては，リノール酸に導入された不安定なヒドロペルオキシ基をパーアセタールとしての保護する事により解決した．この保護基は，中間体から最終生成物に至るまでの反応条件，例えばDCCによるアシル化反応に対して安定である事が明らかとなった．この二つの問題を解決する事によって，リン脂質過酸化物の一つであるホスファチジルコリン過酸化物を世界に先駆けて成功した．さらにこのホスファチジルコリン過酸化物に微生物由来のホスフォリパーゼDを作用させる事によってホスファチジルエタノールアミン過酸化物，ホスファチジルセリン過酸化物やホスファチジルグリセロール過酸化物の合成にも成功した．また，トリグリセリド過酸化物の合成も可能になった．これらの脂質過酸化物が化学的に実態のあるものとして認識されてから，その生理作用に関する研究が広範に行われている．しかし，生体組織に存在するリン脂質過酸化物の生理学的役割は依然として明らかになっていない．ある種のリン脂質過酸化物が動物の免疫系を活性化するという報告もあり，必ずしも生体に対して悪い作用をするばかりではないようである

Determination of a phosphorylation site in Nipah virus nucleoprotein and its involvement in virus transcription

Many viruses use their host’s cellular machinery to regulate the functions of viral proteins. The phosphorylation of viral proteins is known to play a role in genome transcription and replication in paramyxoviruses. The paramyxovirus nucleoprotein (N), the most abundant protein in infected cells, is a component of the N–RNA complex and supports the transcription and replication of virus mRNA and genomic RNA. Recently, we reported that the phosphorylation of measles virus N is involved in the regulation of viral RNA synthesis. In this study, we report a rapid turnover of phosphorylation in the Nipah virus N (NiV-N). The phosphorylated NiV-N was hardly detectable in steady-state cells, but was detected after inhibition of cellular protein phosphatases. We identified a phosphorylated serine residue at Ser451 of NiV-N by peptide mass fingerprinting by electrospray ionization–quadrupole time-of-flight mass spectrometry. In the NiV minigenome assay, using luciferase as a reporter gene, the substitution of Ser451 for alanine in NiV-N resulted in a reduction in luciferase activity of approximately 45 % compared with the wild-type protein. Furthermore, the substitution of Ser451 for glutamic acid, which mimics a phosphoserine, led to a more significant decrease in luciferase activity – approximately 81 %. Northern blot analysis showed that both virus transcription and replication were reduced by these mutations. These results suggest that a rapid turnover of the phosphorylation of NiV-N plays an important role in virus transcription and replication

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DRFIC2008 Poster Session Recap one-minute oral presentation No.1DRFIC2008 ポスターセッションまとめ 口頭プレゼンテーション資料 1

Calreticulin and integrin alpha dissociation induces anti-inflammatory programming in animal models of inflammatory bowel disease

Inflammatory bowel disease (IBD), including ulcerative colitis and Crohn’s disease, is a chronic intestinal inflammatory condition initiated by integrins-mediated leukocyte adhesion to the activated colonic microvascular endothelium. Calreticulin (CRT), a calcium-binding chaperone, is known as a partner in the activation of integrin α subunits (ITGAs). The relationship between their interaction and the pathogenesis of IBD is largely unknown. Here we show that a small molecule, orally active ER-464195-01, inhibits the CRT binding to ITGAs, which suppresses the adhesiveness of both T cells and neutrophils. Transcriptome analysis on colon samples from dextran sodium sulfate-induced colitis mice reveals that the increased expression of pro-inflammatory genes is downregulated by ER-464195-01. Its prophylactic and therapeutic administration to IBD mouse models ameliorates the severity of their diseases. We propose that leukocytes infiltration via the binding of CRT to ITGAs is necessary for the onset and development of the colitis and the inhibition of this interaction may be a novel therapeutic strategy for the treatment of IBD