[Approaches for Class Preparation Using Qualitative Analysis by Coding-Survey in Technical English Course "Yakugaku-Eigo Nyumon"].

The purpose of this study was to examine how students prepare for the pharmaceutical technical English course "Yakugaku-Eigo Nyumon" by qualitative analysis. A sub-text, supplemental material was used to assist students with class preparation. Qualitative questionnaires on understanding and approaches for class preparation as well as review of class were analyzed in comparison with different academic performance levels on the final exam. The results of qualitative analysis of class preparation based on coding revealed that high-academic-performing students understood and adopted deep-processing approaches for the preparation of "English words" and "understanding of content" more often than low-academic-performing students. High-performing students attempted to not literally translate English sentences into Japanese while checking the English words with thinking and ingenuity, and to understand English sentences by drawing figures and thinking of relationships using previously learned knowledge. These approaches were not adopted by low-performing students. Furthermore, sub-text was one of the means for understanding by high-performing students, whereas it was essential for low-performing students to understand the content. Coding results on the review of class also showed that low-performing students were dependent mainly on sub-text for understanding. These results suggest that deep-processing approaches to both English and content of materials are necessary for deep understanding in "Yakugaku-Eigo Nyumon".

Enhancing Study Motivation and Efficacy among First-year Students Using Minute Papers in the Interdisciplinary Subject of Yakugaku Nyumon.

Active learning in higher education is important for learning efficacy and motivation. Accordingly, lectures that integrate strategies toward active learning, such as minute papers, debates, and collaborative learning, have become widely adopted. Minute papers facilitate communication among both teachers and students, and can be used as a tool for reviewing lectures. In the present study, we examined the effect of using minute papers on learning efficacy and motivation. To enhance the curriculum of the interdisciplinary course Yakugaku Nyumon, which consists of an omnibus lecture series and problem-based learning, minute papers with exercises were provided to applicants. In a follow-up questionnaire, students who used minute papers (S-USE) responded that they had a better understanding of the relationships, ranging from basic to clinical subject matter, than students who did not use such papers (S-NON). Using the Attention, Relevance, Confidence, and Satisfaction (ARCS) model questionnaire to measure study motivation, S-USE scored higher for some questionnaires than S-NON. This finding indicates that minute papers promoted learning motivation among students taking the Yakugaku Nyumon course. In regular examinations, the average score of S-USE was also statistically higher than that of S-NON. These results demonstrate that minute papers possibly encouraged students to actively review the lectures, thereby increasing both learning efficacy and motivation. This study shows that through promoting active, self-learning, minute papers are suitable for improving curricular strategies in subjects that rely on passive learning methods.

[Jigsaw Method Is Used to Promote a First Year-student's Understanding of Integrated Subjects at Kobe Pharmaceutical University].

In the School of Pharmacy, collaborative learning that includes both problem-based learning (PBL) and team-based learning (TBL) methods, has been introduced as an active learning method into the education of first-year pharmacy students. These methods are an educational approach to teaching and learning that involve groups of students working collaboratively to resolve a problem, complete a task, or develop a product. However, these methods might not aid students who focus more on the results than on the problem-solving process. In addition, the self-efficacy and learning motivation of students who dislike these learning methods might decrease. The Jigsaw Method respects the individuality of students and is a collaborative learning approach that decreases conflict among students with varied learning styles. Upon applying this method in the first-year course at Kobe Pharmaceutical University, it was observed that most students who attended the life science class increased their self-efficacy, and their passive learning attitudes transformed into active ones. The introduction of the Jigsaw Method to the education of first-year students can help them acquire an effective technique for learning integrated subjects.

[Trial and evaluation of a remedial education program at Kobe Pharmaceutical University].

Pharmaceutical education is suffering from the decentralization of students due to the establishment of new pharmaceutical universities, the shift to a six-year program from a four-year program in the colleges of pharmacy in 2006, and a decrease in the number of students. Combined, these have lead to academic failings, even at high-ranking universities. However, abundant knowledge and ability are necessary to pass the national examination for pharmacists. At Kobe Pharmaceutical University, a Basic Education Center for Pharmacy was instituted in 2006 for the purpose of supporting the scholastic abilities of students with various challenges, as well as for students in general. One approach at the Basic Education Center for Pharmacy, has been to offer supplementary lessons, and to provide additional opportunities for learning for the repeater. We offer supplementary lessons as an "Office Hours Class" and also use DVD-learning for remedial teaching. "Office Hours Class" is conducted in a question/answer format with a small number of students. In order to develop the DVD-learning system (pharmaceutical educational digital learning; PEDL), which promotes self-learning, our supplementary lectures and "Office Hours Class" lectures are recorded and edited on DVD media. The learning effect of using these systems, as determined by regular examination, shows that these have been helpful. As a result, we concluded that these supplementary learning programs are useful as a learning method to help students acquire necessary knowledge as potential pharmaceutical professionals, and to increase student motivation.

Simultaneous determination of histamine and prostaglandin D2 using an LC-ESI-MS/MS method with positive/negative ion-switching ionization modes: application to the study of anti-allergic flavonoids on the degranulation of KU812 cells.

A new method for simultaneous determination of histamine and prostaglandin D(2) (PGD(2)) by liquid chromatography-electrospray ionization tandem mass spectrometry operated in positive and negative ionization switching modes was developed and validated without a previous derivatization step. This method was used to measure histamine and PGD(2) release following degranulation of KU812 human basophilic cells, using pyrazol and d(4)-PGD(2) as internal standards. Analyses were performed on a liquid chromatography system employing a Cosmosil 5C(18) PAQ column and an isocratic elution with mixed solution of methanol-water (7:3, v/v) with 0.0015% trifluoroacetic acid at a flow rate of 0.2 mL/min. A triple-quadrupole mass spectrometer operating in selected reaction monitoring mode simultaneously monitored using the following transitions: positive m/z 112/95 for histamine and negative m/z 351/271 for PGD(2). The retention times of histamine and pyrazol were 4.2 and 5.0 min, respectively. PGD(2) and d(4)-PGD(2) had retention times of 8.5 min. The limits of detection were 0.3 and 0.5 ng/mL for histamine and PGD(2), respectively. The relative standard deviations of the retention time and peak area for histamine were between 1.6% and 7.7%, and were 1.2% and 7.8% for PGD(2). This method was used to evaluate the anti-allergic effects of 26 flavonoids and sodium cromoglicate which are first-line anti-allergic drugs. Of these compounds, baicalein and morin were the most potent inhibitors.

Correlation between cytotoxic activities and reduction potentials of heterocyclic quinones.

To search for possible anti-tumor agents or anti-tumor promoters among natural or synthetic products, we used cyclic voltammetry to determine the reduction-oxidation potentials of heterocyclic quinones in phosphate buffer at pH 7.2. We determined the growth inhibitory- and cytotoxic activities of 12 heterocyclic quinone anti-tumor agent candidates against a panel of 39 human cancer cell lines (JFCR39). The average concentrations of the heterocyclic quinones required for 50% growth inhibition (GI(50)) against JFCR39 ranged from 0.045 to 13.2 µM, and the 50% lethal concentration (LC(50)) against JFCR39 ranged from 0.398 to 77.7 µM. The average values of GI(50) or LC(50) of the heterocyclic quinones correlated significantly with their reduction potentials. These results suggested that reduction-oxidation potentials could be a useful method for the discovery of novel antitumor agents.

Biotransformation of phenolic tetrahydroprotoberberines in plant cell cultures followed by LC-NMR, LC-MS, and LC-CD.

A metabolic pathway of 2,3,10,11-oxygenated tetrahydroprotoberberines having the OH group on ring D was demonstrated. Metabolism of (13)C- or D(2)-labeled precursors was studied in cell cultures of Macleaya, Corydalis, and Nandina species. The structures of alkaloid metabolites obtained from feeding experiments were determined by application of combined LC-NMR, LC-MS/MS, and LC-CD techniques. (S)-Tetrahydropseudoprotoberberine (5) was stereospecifically O-methylated to the S-isomer (12) in cell cultures of three plant species. This S-isomer was further N-methylated to the (S)-alpha-N-methyl salt (15), which was oxidized to produce the pseudoprotopine-type alkaloid (10) in cell cultures of Macleaya and Corydalis species. These transformations were the same as those of 2,3,9,10-oxygenated protoberberines. The tetrahydropseudoprotoberberines (5, 6, and 12) were dehydrogenated to pseudoprotoberberines (13, 16, and 14), respectively. Both the R- and S-enantiomers of 5 were dehydrogenated in Macleaya cordata different from the case of 2,3,9,10-oxygenated protoberberines. Precursor 7, with OH groups at C-10 and C-11, was O-methylated at C-10 in M. cordata and C. ochotensis var. raddeana, which was distinct from O-methylation in N. domestica, in which 7 was O-methylated at both C-11 and C-10. Stereoselective O-demethylation [(S)-5 to (S)-18] occurred in N. domestica.

Characterization of emodin metabolites in Raji cells by LC-APCI-MS/MS.

A rapid, simple, and sensitive liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry (LC-APCI-MS/MS) method was developed for the identification and quantification of emodin metabolites in Raji cells, using aloe-emodin as an internal standard. Analyses were performed on an LC system employing a Cosmosil 5C(18) AR-II column and a stepwise gradient elution with methanol-20mM ammonium formate at a flow rate of 1.0 mL/min operating in the negative ion mode. As a result, the starting material emodin and its five metabolites were detected by analyzing extracts of Raji cells that had been cultivated in the presence of emodin. The identification of the metabolites and elucidation of their structures were performed by comparing their retention times and spectral patterns with those of synthetic samples. In addition to the major metabolite 8-O-methylemodin, four other metabolites were assigned as omega-hydroxyemodin, 3-O-methyl-omega-hydroxyemodin, 3-O-methylemodin (physcion), and chrysophanol.

Development of an LC-ESI-MS/MS method for the determination of histamine: application to the quantitative measurement of histamine degranulation by KU812 cells.

A rapid, simple, and sensitive liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) method was developed for the identification and quantification of histamine without a previous derivatization step or the addition of general ion-pairing reagents to the mobile phase. This method was used to measure histamine release following degranulation of KU812 human basophilic cells, using pyrazol as an internal standard. Analyses were performed on an LC system employing a Cosmosil 5C(18) PAQ column and an isocratic elution with methanol-0.005% trifluoroacetic acid (1:1) at a flow rate of 0.2 mL/min. A triple-quadrupole mass spectrometer, equipped with an electrospray ionization interface was employed, operating in the positive ion mode. The retention time of histamine and the internal standard were 4.0 and 5.0 min, respectively. The relative standard deviations (R.S.D.s) of the retention time and peak area were between 0.47% and 2.03%. Micropipette tip solid-phase extraction (SPE) using LooseTip C(18) allowed for not only rapid sample preparation, but also decreased suppression effects, improving peak shape. This method was used to evaluate the anti-allergic effects of compounds contained in Taxus yunnanensis extracts. Four constituents that were isolated from the wood extracts of T. yunnanensis and sodium cromoglicate, which is used as a first line anti-allergic drug, were tested in an in vitro histamine release inhibition assay. Of these compounds, taxiresinol and isotaxiresinol were more inhibitory than sodium cromoglicate.

Correlation between reduction potentials and inhibitions of Epstein-Barr virus activation by anthraquinone derivatives.

As a continuation of studies using natural and synthetic products as cancer chemopreventive agents, we used cyclic voltammetry to examine the reduction-oxidation potentials of methylated emodin derivatives prepared from emodin in phosphate buffer at pH 7.2. A good correlation was found between the inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation and the reduction potential of methylated emodin derivatives. Furthermore, there was significant correlation between EBV-EA activation and the reduction potential of 35 anthraquinone derivatives including methylated emodin derivatives. It was further shown that the correlation could be enhanced by including LUMO energy and the number of hydroxy groups as additional parameters.

Correlation between oxidation potentials and inhibitory effects on Epstein-Barr virus activation of flavonoids.

The oxidation potentials of fifteen flavonoids in phosphate buffer at pH 7.2 were determined by cyclic voltammetry. A good correlation was found between these oxidation potentials and the ability of flavonoids to inhibit Epstein-Barr virus early antigen (EBV-EA) activation. Furthermore, multiple regression analysis revealed that the solvent-accessible surface area (SASA) was a useful parameter for estimating the inhibitory effects of flavonoids on EBV-EA activation.

Simultaneous determination of anthraquinones in rhubarb by high-performance liquid chromatography and capillary electrophoresis.

High-performance liquid chromatography (HPLC) and capillary electrophoresis (CE) were compared to simultaneously determine and separate 11 anthraquinones from rhubarb, including emodin, chrysophanol, rhein and their glucosides, aloe-emodin, sennoside A, and sennoside B. A UV-diode array detector (DAD) at 254 nm with a gradient elution of acetonitrile/water (method A: 0 min 6:94, 12 min 12:88, 15 min 20:80, 40 min 25:75, 53 min 55:45, 55 min 100:0; method B: 0 min 5:95, 2 min 15:85, 5 min 20:80, 12 min 25:75, 15 min 50:50, 19 min 98:2) at 28(+/-1) degrees C (method A) and 30-60 degrees C (method B) in HPLC or with 0.03 M borate buffer (pH 10.0) containing 25% (v/v) acetonitrile with 0.002 M 2,6-di-O-methyl-beta-cyclodextrin (CD) and 0.005 M alpha-CD in CE effectively detected this separation in 25 min. The detection limits of anthraquinones from rhubarb were in the 0.02-0.2 microg/mL and 0.1-0.8 microg/mL ranges for HPLC and CE, respectively. The established HPLC and CE methods are suitable for quantitative determination of emodin, chrysophanol, aloe-emodin, emodin-1-beta-d-glucoside, emodin-8-beta-d-glucoside, chrysophanol-1-beta-d-glucoside, chrysophanol-8-beta-d-glucoside, and rhein-8-beta-d-glucoside.

A new quassinoid, ailantinol H, from Ailanthus altissima.

A new quassinoid, ailantinol H, was isolated from the aerial parts of Ailanthus altissima. The structure was elucidated based on spectral evidence.

Antiallergic activity of aqueous extracts and constituents of Taxus yunnanensis.

The H2O, H2O/MeOH (1:1) extracts from the wood of Taxus yunnanensis showed a remarkable inhibitory effect on induced histamine release from the human basophilic cell line, KU812. The eleven constituents purified from the wood extracts of Taxus yunnanensis were tested by an in vitro histamine release inhibition assay. Among them, secoisolarciresinol and taxiresinol were found to show inhibitory activities. A new neolignan, 2-[2-hydroxy-5-(3-hydroxypropyl)-3-methoxyphenyl]-1-(4-hydroxy-3-methoxyphenyl)propane-1,3-diol, was isolated from the wood of Taxus yunnanensis.

Furanonaphthoquinones cause apoptosis of cancer cells by inducing the production of reactive oxygen species by the mitochondrial voltage-dependent anion channel.

The mitochondrial production of reactive oxygen species has been implicated in the anticancer activity of furanonaphthoquinone. However, the mechanism of the activation remains elusive. In the current study, we found that treatment of HeLa cells with 2-methyl-5(or -8)-hydroxy-furanonaphthoquinone (FNQ13) induces mitochondrial swelling, followed by apoptosis. This toxic effect of FNQ13 was reduced by the radical scavengers alpha-tocopherol and trolox. Cytochemical experiments in isolated mitochondria showed that a combination of FNQ13 and NADH induces the production of H(2)O(2) at the exterior mitochondrial membrane surface. This production of H(2)O(2) was reduced by an antibody to the voltage-dependent anion channel (VDAC). Overexpression of the VDAC by transfection with vdac1 cDNA increased the production of H(2)O(2) by HeLa cells, whereas transfection with a small interfering RNA to VDAC reduced FNQ13-induced H(2)O(2) production and cell death due to an almost complete knockdown of VDAC expression. We also found significant correlations between the expression of VDAC and the induction of H(2)O(2) production and cell death by FNQ13 in 11 human cancer cell lines. These results indicate that the anticancer activity of furanonaphthoquinones depends on the production of reactive oxygen species by mitochondrial permeability transition pores including the VDAC.

Correlation between reduction potentials and inhibitory effects on Epstein-Barr virus activation by emodin derivatives.

As a continuation of studies using natural and synthetic products as cancer chemopreventive agents, we examined the reduction-oxidation potentials of hydroxylated emodin derivatives prepared from emodin in phosphate buffer at pH 7.2 using cyclic voltammetry. A significant correlation was found between the reduction potentials and number of the hydroxyl groups and the inhibitory effects of the hydroxylated emodin derivatives on Epstein-Barr virus early antigen activation. The electronic properties, i.e. LUMO energy and atomic charges of carbon at the 9-position (C(9)) and oxygen at the 11-position (O(11)), may also be useful for estimating the inhibitory effect on EBV-EA activation.

Anti-infective quinone derivatives of recent patents.

Quinones are important naturally occurring pigments widely distributed in nature and are well known to demonstrate various physiological activities as antimicrobial and anticancer compounds. This review will focus on the preparation, therapeutic application, and administration of several benzoquinones, naphthoquinones, and anthraquinones having anti-infective, e.g. antiviral and antibacterial activities, in recent patents.

Correlation between reduction potentials and inhibitory effects on Epstein-Barr virus activation of poly-substituted anthraquinones.

As a continuation of our studies using natural and synthetic products as cancer chemopreventive agents, we examined the reduction potentials of some poly-substituted anthraquinones in phosphate buffer at pH 7.2 by means of cyclic voltammetry. A definite correlation has been found between the reduction potentials and the inhibitory effects of the poly-substituted anthraquinones on Epstein-Barr virus early antigen activation. It has been further shown that the correlation can be enhanced by introducing log P as an additional parameter.

Simultaneous determination of anthraquinones, their 8-beta-D-glucosides, and sennosides of Rhei Rhizoma by capillary electrophoresis.

The simultaneous separation and determination of major anthraquinones (emodin, chrysophanol, rhein and their glucosides, aloe-emodin, sennoside A, and sennoside B) of Rhei Rhizoma were achieved by cyclodextrin modified capillary zone electrophoresis. The running electrolyte used in this method was 0.005 M alpha-cyclodextrin in 0.03 M borate buffer (pH 10.0) containing 20% acetonitrile, with an applied voltage of 20 kV.

Structure-activity relations of azafluorenone and azaanthraquinone as antimicrobial compounds.

Antimicrobial activities of two azafluorenones, four 1-azaanthraquinones, five 2-azaanthraquinones, and one 2-azaquinone were tested. Several azaanthraquinones possessed broad, potent activity, while the azafluorenones demonstrated weak activity. The following structure-activity relationship was postulated: (1) activity decreased in the order 2-azaanthraquinones>1-azaanthraquinones>azafluorenones; and (2) a hydroxyl group at the peri-carbonyl group enhanced activity. In addition, correlations among reduction potential, hydrophobic parameter, and antimicrobial activity were discussed.