西丸 宏

武蔵野大学薬学部6 年生に対して実施した薬剤師国家試験と同形式の試験（9 月～2 月に計6 回実施）における総合計および各科目の成績推移をグループ化できるかについて，3 年間（2017–2019 年度）のデータを用いたクラスター分析を行って検討した．その結果，総合計およびすべての科目について上位・中位・下位の3 グループに分類することができたが，その構成人数は科目によって異なる傾向を示した．また，すべての科目について総合計との相関が見られた．ある時点の試験成績から薬剤師国家試験の合否に関して有意に影響がある科目を見いだせるかについて多重ロジスティック回帰分析で検討したところ，薬理など複数の科目で合否に影響がある可能性が示唆された．本研究は，各学生の早い時期の試験成績から，学習到達度の把握や今後の成績推移の見通しを予測し，国家試験対策における学生指導や学習方略の構築を検討する際に有用な知見となる可能性を示している

本学ではアクティブ・ラーニングの一環として学外学修プログラムに取り組んでおり、薬学部以外の1年生は必修としている。プログラムの一つにロサンゼルスの医療福祉施設視察研修があり、看護学部や人間科学部の学生が参加している。薬学部生は履修制度の関係で単位は取得できないが、希望者は本プログラムに参加可能となっている。今回、2016年度の本プログラムに参加した学生が終了後に提出したレポートの自由記述内容をテキストマイニングで分析し、キーワードの抽出を行った。結果、出現頻度が最も高いワードは【学ぶ・知る】であり、次いで【日本】、【病院・施設】【アメリカ】、【自分】、【多い・たくさん】、【見る・見学】、【質問】、【働く・仕事】、【違い】、【コミュニケーション】の順であった。

This study examined the pharmacokinetics and pharmacological activities of 6-shoganol, a pungent ingredient of Zingiberis Rhizoma, and its metabolite, 6-paradol. The concentrations of 6-shogaol and 6-paradol in rat plasma determined by LC/MS/MS reached their maximum values (Cmax) at 5 minutes after oral administration of 6-shogaol (10 mg/kg). Both 6-shogaol and 6-paradol were eliminated from the plasma within 2 hours after injection. The plasma concentration of 6-paradol, the metabolite, was about 4 times higher than that of 6-shogaol at all points during blood sampling. Next, pharmacological activities of 6-shogaol and 6-paradol were studied. In vitro experiment revealed that the cyclooxygenase-2-inhibitory activity of 6-paradol was about 6 times stronger than that of 6-shogaol. In vivo experiments, 6-paradol demonstrated significantly stronger anti-inflammatory, analgesic, and antipyretic activities compared to 6-shogaol. These results suggest that 6-shogaol in Zingiberis Rhizoma is metabolized rapidly to 6-paradol and that 6-paradol is the main compound having anti-inflammatory activity. © 2013, Medical and Pharmaceutical Society for WAKAN-YAKU. All rights reserved.

Five phenylethanoid glycosides (acteoside, isoacteoside, beta-oxoacteoside, beta-hydroxyacteoside, and salidroside) were isolated from a cell suspension culture of Olea europaea. We examined the biosynthesis of acteoside in olive cell cultures by using feeding experiments with stable isotope labeled precursors. The hydroxytyrosol moiety of acteoside is biosynthesized from tyrosine through dopamine, whereas the caffeoyl moiety of acteoside is biosynthesized from phenylalanine via a cinnamate pathway. Dopamine is incorporated into acteoside through oxidation to the corresponding aldehyde, reduction to the alcohol, and then beta-glycosylation.

It has been known that the over-expression of alpha-synuclein, the main protein of Lewy bodies in Parkinson's disease (PD), leads to neurodegeneration in PD models. In this study, the changes in protein expression between the transgenic over-expressing human alpha-synuclein wild type (alpha-synWT) and the control Caenorhabditis elegans were elucidated by fluorogenic derivatization-liquid chromatography/tandem mass spectrometry (FD-LC-MS/MS) proteome analysis, which is a highly selective, sensitive, repeatable and quantitative method for protein identification. Because the a-synuclein wild-type worms showed moderate levels of dopamine loss without overt behavioral abnormalities, it was suggested that the changes in proteins in the alpha-asynWT are related in the sequence of the formation of Lewy bodies. Among more than 400 protein peaks detected, actin and several ribosomal proteins were identified for the first time as negative markers at early PD stages. Actin was suggested to be one of the important targets in the elucidation of the etiology of neuronal diseases such as PD or other synucleinopathies. Copyright (c) 2007 John Wiley & Sons, Ltd.

Olive (Olea europaea) contains large quantity of triterpene acids including oleanollic acid (6) as a major one. Varieties of biological activities exhibited by triterpene acids attracted our attentions, especially from pharmaceutical viewpoints. Cell culture of olive plant was induced and its triterpene constituents were studied. From the cell suspension cultures, six ursane type triterpene acids; ursollic acid (9), pomolic acid (10), rotundic acid (11), tormentic acid (12), 2 alpha-hydroxyursolic acid (13) and 19 alpha-hydroxyasiatic acid (14), and two oleanane type acids; oleanolic acid and maslinic acid (7), have been isolated. Quantity of ursane type triterpene acids produced by cell cultures was larger than that of oleanane type. Further, a multifunctional oxidosqualene cyclase (OSC) named OEA was cloned by homology based PCRs from the same cultured cells. Major product of OEA is alpha-amyrin (ursane skeleton), showing good accordance to higher content of ursane-type triterpene acids in the cultured cells, and strongly suggesting OEA to be a major contributor OSC for their production.

To find the pairs of fluorogenic reagents having similar retention times in HPLC but with different fluorescent characteristics, six fluorogenic reagents bearing benzoxadiazole or benzoselenadiazole skeletons were synthesized. The resultant derivatives obtained from the reaction of peptides and proteins with reagents which have a benzoselenadiazole skeleton showed different fluorescence characteristics from those with a benzoxadiazole skeleton. Since each corresponding derivatives of trypsin inhibitor and BSA with DAABD-Cl and 7-fluoro-N-[2-(diethylamino)ethyl]-2,1,3-benzoselenadiazole-4-sulfonamide (DEAEABSeD-F) have similar retention times, the pair of reagents was adopted for the sensitive simultaneous detection of proteins in two different samples. When the soluble fraction of mouse hippocampus was divided into the two samples (A and B), each was reacted with DEAEABSeD-F for A and DAABD-Cl for B, respectively. The two reaction solutions were combined and subjected to HPLC analysis with two fluorescent detectors in series (excitation and emission at different wavelengths for A and B, respectively). The resultant two chromatograms had quite similar patterns for each other. The new pair of fluorogenic reagents (DAABD-Cl and DEAEABSeD-F) would be applicable to proteomics studies using the previously reported FD-LC-MS/MS method. Copyright (c) 2006 John Wiley & Sons, Ltd.

The derivatization regents for carboxylic acids, DAABD-AE {4-[2-(N,N-dimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole}, MePZBD-AE {[4-(4-N-methyl)piperazinosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole} and APZBD-NHMe {[4-(4-N-aminoethyl)piperazinosulfonyl]-7-methylamino-2,1,3-benzoxadiazole} were developed for electrospray ionization-mass spectrometry (ESI-MS). The derivatization reaction with fatty acids was completed at 60 degrees C within 30 min. The derivatives of fatty acids were separated on a reversed-phase column and detected with ESI-MS. The detection limits attained for fatty acids were femtomol range and the calibration curves were linear over the range from 0.1 to 100 pmol (r(2) > 0.992) for DAABD-AE and MePZBD-AE. DAABD-NHMe was applied to the analysis of fatty acids in rat plasma samples. Copyright (c) 2005 John Wiley & Sons, Ltd.

An improved method for proteomics studies, which includes the fluorogenic derivertization of protein mixtures with 7-chloro-4-(dimethylaminoethylaminosulfonyl)-2,1,3-benzoxadiazole (DAABD-Cl), followed by HPLC isolation, enzymatic digestion and idetification of the derivatized proteins by HPLC-electrospray ionization (ESI)-MS/MS with the probability-based protein identification algorithm, identified 103 proteins in the soluble extract (10 mu g protein) of Caenorhabditis elegans. Copyright (c) 2005 John Wiley & Sons, Ltd.

化学発光反応は蛍光よりも高い感度を持ち検出反応に利用されている。本稿では脳髄液物質の化学発光反応による定量法について概説する。

Previously, we have synthesized a thiol specific fluorogenic reagent, 7-chloro-N-[2(dimethylamino)ethyl]-2,1,3-benzoxadiazole- 4-sulfonamide (DAABD-Cl) and derivatized protein mixtures, isolated the derivatives by HPLC, followed by enzymatic digestion and identification of the derivatized proteins by HPLC-electrospray ionization mass spectrometry (ESI-MS/MS) with the probability-based protein identification algorithm (FD-LC-MS/MS method) ( M. Masuda et al., Anal. Chem., 2004, 76, 728). The emission wavelengths for the DAABD derivatives of proteins were 510 nm with an excitation wavelength of 380 nm. In this paper, a new thiol specific fluorogenic reagent, 7-fluoro-N-[2-(dimethylamino)ethyl]-2,1,3-benzoselenadiazole-4-sulfonamide (DAABSeD-F) was synthesized. It reacted with thiols of peptides and proteins to give fluorescence. The respective excitation and emission wavelengths of the DAABSeD derivatives of proteins were about 30 nm and 40 nm red shifted from those of the DAABD proteins. These properties would be appropriate for its use in combination with DAABD-Cl as a counterpart for the simultaneous detection, ratio calculation and identification of peptides or proteins produced under the different conditions. Thus, DAABSeD- F could be an additional supportive reagent for the FD-LC-MS/MS method.