Direct evidence of specific localization of sesquiterpenes and marchantin A in oil body cells of Marchantia polymorpha L.

Liverworts are a rich source of a diverse array of specialized metabolites, such as terpenoids and benzenoids, which are potentially useful for pharmaceutical or agrochemical applications, and also provide clues to elucidate the strategy by which liverworts adapt to the terrestrial environment. Liverworts, belonging to orders Marchantiales and Jungermanniales, possess oil bodies. In Marchantia polymorpha L., oil bodies are confined to scattered idioblastic oil body cells. It has been assumed that the specialized metabolites in M. polymorpha specifically accumulate in the oil bodies in oil body cells; however, no direct evidence was previously available for this specific accumulation. In this study, direct evidence was obtained using micromanipulation techniques coupled with MS analysis that demonstrated the specific accumulation of sesquiterpenoids and marchantin A in the oil body cells of M. polymorpha thalli. It was also observed that the number of oil body cells increased in thalli grown in low-mineral conditions. The amounts of sesquiterpenoids and marchantin A detected in crude extract prepared from the whole thallus were roughly proportional to the number of oil body cells found in a given volume of thallus, suggesting that oil body cell differentiation and sesquiterpenoid and marchantin A biosynthetic pathways are coordinated with each other.

Bioactivity guided isolation of anticancer constituents from leaves of Alnus sieboldiana (Betulaceae).

The leaves of the Japanese Alnus sieboldiana have been extracted with n-hexane and then with methanol. A bioactivity-guided approach based on MTT assay for growth inhibition and quantitative real-time PCR for TNF-α inhibitory activity was taken to identify the active compounds in EtOAc soluble fraction of the methanol extract. From this active fraction, seven compounds have been isolated and four compounds (pinosylvin, galangin, quercetin and methyl gallate) have been examined for their dose-response effect on the viability of A549 cells and on TNF-α inhibitory activity. Based on MTT assay, all of the four examined compounds inhibit growth of human lung cancer cells. Among four tested compounds only galangin (3,5,7-trihydroxyflavone) significantly inhibited TNF-α gene expression in A549 cells (IC50 = 94 µM). Taken together, this finding suggests that galangin may be useful in cancer prevention.

Introduction of community-based education for physical therapy students in Japan / Монголын Анагаах Ухаан

Background: Physical therapists need to work not only in hospitals but also in the communities. So it has been said that community-based education should be essential for physical therapy education. In Gunma University, community-based education program was implemented from 2010 for physical therapy stu¬dents. Purpose. The purpose of this report is to introduce our challenging program and search for how this program af¬fected students, community staffs and elderly people. Methods: Third-year students collaborated to exercise classes for community dwelling elderly people with teach¬ers and community staffs (public health nurses and civic volunteers). Two classes were provided separately, so students were divided into two groups and each group was assigned to one class. After classes, reports were written by participants (60 persons), community staffs (9 persons) and students (24 persons). Reports consisted of five-grade evaluation of this program and students for elderly people and community staffs, evaluation using visual analogue scale (VAS) of this program and contact with elderly people for students, and impressions described in freely written sentences for all subjects. Results and Discussion: Results of elderly people’s five-grade evaluation were “5 (very good)”-50 persons, “4 (good)”-10 persons to this program and “5”-41 persons, “4”-19 persons to students. Results of community staffs’ five-grade evaluation were “5”-7 persons, “4”-2 persons to this program and “5”-6 persons, “4”-3 persons to stu¬dents. Results of students’ VAS evaluation were 86.5±13.3 (100 extremely good~0 minimum) to this program and 91.6±9.9 to contact with elderly people. It was expected there were good feelings elderly people, community staffs and students have. There was a possibil¬ity that an interchange between elderly people and students would bring different learning from on-campus classes to students and motivation to elderly people. Take-home message: It is thought that community-based education should be included in undergraduate pro¬grams for physical therapy students.

Liverworts-potential source of medicinal compounds.

The bryophytes contain the Marchantiophyta (liverworts), Bryophyta (mosses) and Anthocerotophyta (hornworts) among which the Marchantiophyta contain cellular oil body and they produce a number of terpenoids, aromatic compounds and acetogenins, several of which show interesting biological activity such as allergenic contact dermatitis, insecticide, insect antifeedant, cytotoxic, piscicidal, muscle relaxing, plant growth regulatory, anti-HIV, DNA polymerase beta inhibitory, anti-obesity, neurotrophic, NO production inhibitory, antimicrobial and antifungal activities. The isolation and chemical structures of biologically active compounds and their total synthesis are reviewed.

Screening of novel nuclear receptor agonists by a convenient reporter gene assay system using green fluorescent protein derivatives.

Nuclear receptors represent a very good family of protein targets for the prevention and treatment of diverse diseases. In this study, we screened natural compounds and their derivatives, and discovered ligands for the retinoic acid receptors (RARs) and the farnesoid X receptor (FXR). In the reporter assay systems of nuclear receptors presented here, two fluorescent proteins, enhanced yellow fluorescent protein (EYFP) and enhanced cyan fluorescent protein (ECFP), were used for detection of a ligand-based induction and as an internal control, respectively. By optimizing the conditions (e.g., of hormone response elements and promoter genes for reporter plasmids), we established a battery of assay systems for ligands of RARs, retinoid X receptor (RXR) and FXR. The screening using the reporter assay system can be carried out without the addition of co-factors or substrates. As a result of screening of more than 140 compounds, several compounds were detected which activate RARs and/or FXR. Caffeic acid phenylethyl ester (CAPE), known as a component of propolis from honeybee hives, and other derivatives of caffeic acid up-regulated the expression of reporter gene for RARs. Grifolin and ginkgolic acids, which are non-steroidal skeleton compounds purified from mushroom or ginkgo leaves, up-regulated the expression of the reporter gene for FXR.

Isolation and synthesis of TNF-alpha release inhibitors from Fijian kawa (Piper methysticum).

Two unique evidence that cancer incidence rates in Fiji were unusually low, compared with those of another Pacific islands and that green tea beverage is an acknowledged cancer preventive in Japan, allowed us to study a local beverage in Fiji, kawa (kava kava) or yangona (Piper methysticum) belonging to Piperaceae. We isolated five known kawapyrones (kavapyrones) (1-5) and a new additional kawapyrone, 7,8-epoxyyangonin (6), from kawa MeOH extract and subjected them to TNF-alpha (tumor necrosis factor-alpha) release assay from BALB/3T3 cells treated with okadaic acid, a tumor promoter. 5,6-Dehydrokawain (desmethoxyyangonin)(1) and yangonin (4) significantly inhibited TNF-alpha release with IC50 values of 17 microM and 40 microM; a potency as great as (-)-epigallocatechin gallate (EGCG) isolated from green tea extract. Among the experiments with 1-5, dihydrokawain (2) was unique in showing the strongest inhibitory activity against TNF-alpha release in mice, but the weakest activity in the cells. We synthesized 5,6-dehydrokawain (1) and yangonin (4) via three steps from the dianion of ethyl acetoacetate achieving a good yield and determined their conformations by high resolution NMR and x-ray crystallographic analysis.

High-sensitivity negative-ion laser spray for liquid chromatography/mass spectrometry.

A negative-ion mode laser spray interface for use in liquid chromatography/mass spectrometry (LC/MS) has been developed and investigated. The laser spray gave sensitivities that were orders of magnitude better than the negative-ion electrospray for all samples investigated with the exception of sugars. Fragment ions, HSO(-4)and SO(-3), were formed from the laser-sprayed aqueous solution of cholesterol 3-sulfate sodium salt. This suggests that structural information may be obtained directly from the laser-spray mass spectra for thermally labile compounds.

Occurrence of polygodial and 1-(2,4,6-trimethoxyphenyl)-but-2-en-1-one from some ferns and liverworts: role of pungent components in bryophytes and pteridophytes evolution.

The New Zealand fern Blechnum fluviatile and liverwort, Hymenophyton flabellatum produce the characteristic pungent compounds, (-)-polygodial, a sesquiterpene dialdehyde, and 1-(2,4,6-trimethoxyphenyl)-but-2-en-1-one, respectively. The former compound has been isolated from the Japanese liverwort, Porella vernicosa complex and the latter one from the Japanese fern, Arachinoides standishii. The occurrence of both compounds in both pteridophytes and bryophytes provides another important link between bryophytes and ferns.

Sesqui- and diterpenoids from the Japanese liverwort Jungermannia infusca.

Seven new cuparane-type (1-7), one new acorane-type (8), one new monocyclic-type (9), and one new prelacinane-type (10) sesquiterpenoid as well as two new clerodane-type (11, 12) and one new halimane-type (13) diterpenoid were isolated from the Japanese liverwort Jungermannia infusca, together with 12 known cuparane- (14-21) and aromadendrane-type (22) sesquiterpenoids and labdane- (23) and ent-kaurane-type (24) diterpenoids. The structures for 1-13 were determined using extensive NMR techniques and by chemical degradation and X-ray crystallographic analysis.

Absolute configuration of (6S,9S)-roseoside from Polygonum hydropiper.

From Polygonum hydropiper L., a C13-norisoprenoid glucoside was isolated and its absolute configuration was established to be (6S,9S)-roseoside (1) by spectroscopic evidence and X-ray crystallographic analysis of its acetate derivative (2). In addition, the stereostructure of roseoside from Canthium subcordatum was revised to the (6S,9S) configuration.

[Falling accidents among metropolitan elderly resulting in emergency ambulance transfer].

The aim of this study was to examine the frequency and number of falling accidents in the the elderly population and to investigate the characteristics of the accidents. The investigation was performed in 58 Tokyo metropolitan townships for a one-year period from September 1st, 1997, and the 517 cases, in which persons were over 65 years old, fell down were studied (136 men, 381 women). The yearly accident frequency for 1,000 elderly over 65 population was 4.13 cases for men and 7.88 cases for women, being 1.9 times higher in women. Also the number of accidents in each township was significantly correlated to the population of the elderly over 65 (men r = 0.674, women r = 0.846, p < 0.001). The analysis by age showed the percentage of accidents in single-family housing among the oldest-old population was significantly higher than the young-old population (p < 0.05) for both men and women. According to logistic regression analysis results, injured body part and the form of dwelling house were significantly correlated as risk factors of fracture (p < 0.05). The odds ratio of injured body part was 7.559 (95% CI 4.926-11.598), and that of the form of dwelling house was 1.660 (95% CI 1.067-2.584), showing that injury of limbs and the accidents in single-family housing would increase risk of fracture. Promoting preventative measures for falling accidents in single-family housing appears important in the prevention of falling accidents among the metropolitan elderly.

New chroman derivatives from the Japanese liverworts Metacalypogeia cordifolia and Cephalozia otaruensis.

The diethyl ether extract of the Japanese liverwort Metacalypogeia cordifolia yielded five new chroman type derivatives in addition to known sesquiterpenoids. One of the new chroman derivatives was also isolated from the ether extract of another liverwort, Cephalozia otaruensis. Their structures were established by extensive two dimensional (2D) NMR techniques and chemical evidence. They were shown to be 2,2-dimethyl-7-(3-methyl-2-butenyl)-chroman derivatives. This was the first example of the isolation of the chroman-type compounds, although various types of aromatic compounds have been isolated from liverworts.

Sesquiterpene hydrocarbons with trifarane backbone in the liverwort Trocholejeunea sandvicensis.

The hydrocarbon fraction of the liverwort Trocholejeunea sandvicensis was investigated. Four new sesquiterpene hydrocarbons including the trifaranes (-)-trifara-9,14-diene (6) and (-)-3,7-di-epi-3,7-trifara-9,14-diene (7), as well as (+)-sandvicene (8), a sesquiterpene with uncommon skeleton, and the rearranged trifarane sesquiterpene (+)-neotrifaradiene (9) were isolated and their structure elucidated. A biogenetic pathway for the four compounds is proposed, with nerolidol as their common precursor.

seco-Cuparane-type sesquiterpenoid from the Japanese liverwort Jungermannia infusca.

A 2,3-seco-cuparane-type sesquiterpenoid and the previously known barbatane-type sesqui- and ent-kaurane-type diterpenoids were isolated from the Japanese liverwort Jungermannia infusca (Mitt.) Steph. The structure of the 2,3-seco-cuparane-type sesquiterpenoid was determined by NMR spectroscopic analyses.

Preparation of gellan sulfate as an artificial ligand for removal of extra domain A containing fibronectin.

The extra domain A containing fibronectin (EDA(+)FN) concentration in plasma of rheumatoid arthritis (RA) is abnormally higher than the normal level. We synthesized various gellan-sulfate (GS) candidates as artificial ligands for removing EDA(+)FN from plasma. The interaction between these artificial ligands and EDA(+)FN was evaluated using affinity constants (KA), which were determined by surface plasmon resonance measurement. The KA (3.6 x 10(8) per M) of GS-25 [degree of substitution for sulfonation (DS) = 25%] with EDA(+)FN was higher than those of other molecules: GS-16 (DS=16%) at 8.3 x 10(7) per M, and GS-35 (DS = 35%) at 1.7 x 10(8) per M. Furthermore, GSs displayed selectivity of EDA(+)FN for binding with plasma FN (KAEDA(+)FN)/KA(plasma FN)>2). The removal ratio in plasma was measured by using GS-immobilized gel. Removals of 66, 11, 7.7, 6.2, 6.9, and 12% for EDA(+)FN, plasma FN, fibrinogen, albumin, immunoglobulin G (IgG) and antithrombin III from the patient-model plasma were, respectively, achieved with GS-25-immobilized gel. These results suggest that GS may be used as a selective artificial ligand for EDA(+)FN removal from plasma in RA treatment.

Pathological fracture through a C-6 aneurysmal bone cyst. Case report.

The authors describe the case of an 18-year-old man who presented with complaints of weakness and paresis in his arms following an injury. Radiological examination demonstrated an aneurysmal bone cyst of C-6. The patient underwent a two-stage operation. Satisfactory results were obtained after complete resection of the lesion, laminoplasty, and anterior fusion without placement of instrumentation. The authors consider a two-stage operation supplemented by fusion without instrumentation to be the best treatment for young patients with aneurysmal bone cysts occurring at C-6.

Sesqui- and diterpenoids from two Japanese and three European liverworts.

A new peroxy muurolane-type sesquiterpenoid was isolated from the ether extract of the Belgium liverwort Scapania undulata, together with three known ent-muurolanes. A new lepidozane-type sesquiterpenoid was isolated from the Japanese Porella subobtusa together with a known santalane- and two africane-type sesquiterpenoids. All structures were determined by means of NMR spectroscopic techniques. The chemosystematics of each species are discussed.

Recent advances in phytochemistry of bryophytes-acetogenins, terpenoids and bis(bibenzyl)s from selected Japanese, Taiwanese, New Zealand, Argentinean and European liverworts.

Bryophytes contain a large number of terpenoids and phenolic compounds. Recent topics relating to the chemical constituents found in 36 Japanese, 3 New Zealand, 2 European, 1 Argentinean and 1 Taiwanese liverworts and 2 Japanese mosses and their biological activity are discussed. The chemosystematics of some liverworts as well as the chemical relationship between liverworts and mosses, and bryophytes and ferns are also discussed.

Causes of airway obstruction during cuffed oropharyngeal airway use.

This study investigated the cause for needing airway maneuvers to maintain a patent airway during the use of cuffed oropharyngeal airway (COPA). Twenty adult patients (29.4+/-6.8 years-old, ASA 1-2) scheduled for minor gynecological surgery who required brief manipulations of the airway despite COPA use following the manufacture's guidelines, were enrolled in this study. To obtain airway patency, 15 patients required only the head-tilt maneuver. In eight of the 15 patients, the laryngeal inlet was opened partially (n=4) or completely (n=4). Despite lifting the epiglottis, the laryngeal inlet was incomplete at the level of pharyngeal view. The patency of the laryngeal inlet was decided by the extent of the distance between the posterior pharyngeal wall and the lateral glossoepiglottic fold, which was made by hyoid bone. In the other seven patients, the head-tilt maneuver elevated the epiglottis and completely opened the laryngeal inlet. Five patients required both the jaw-thrust and head-tilt maneuver. Of these patients lifting the epiglottis was incomplete in three and the laryngeal inlet was partially collapsed in one even after the airway manipulations. The airways in these three patients, however, became patent after manipulations despite the persisting partial obstruction.