Water-soluble constituents from aerial roots of Ficus microcarpa.

Three new water-soluble constituents [ficuscarpanoside B (1), (7E,9Z)-dihydrophaseic acid 3-O-beta-D-glucopyranoside (4) and ficuscarpanic acid (6)] and the natural product 2,2'-dihydroxyl ether (7) have been isolated, together with three known compounds [(7S,8R)-syringoylglycerol (2), (7S,8R)-syringoylglycerol-7-O-beta-D-glucopyranoside (3) and icariside D2 (5)] from the aerial roots of Ficus microcarpa. Identification of their structures was achieved by 1D and 2D NMR experiments, including 1H-1H COSY, NOESY, HMQC and HMBC methods and FAB mass spectral data.

Downregulation of progesterone biosynthesis in rat granulosa cells by adlay (Coix lachryma-jobi L. var. ma-yuen Stapf.) bran extracts.

Adlay (Coix lachryma-jobi L. var. ma-yuen Stapf.) has long been used as a traditional Chinese medicine for dysfunctions of the endocrine system and inflammation conditions. However, the effect of adlay seed on the endocrine system has not yet been reported. In the present study, the effects and the mechanisms of methanolic extract of adlay bran (ABM) on progesterone synthesis in rat granulosa cell were studied. ABM was further partitioned with different solvents including water, 1-butanol, ethyl acetate and n-hexane. Four subfractions named ABM-Wa (water fraction), ABM-Bu (1-butanol fraction), ABM-EA (ethyl acetate fraction) and ABM-Hex (n-hexane fraction) were obtained. ABM-Bu was further fractionated using Diaion HP-20 resin column chromatography with gradient elution. Granulosa cells were prepared from pregnant mare serum gonadotropin-primed immature female rats and challenged with different reagents including human chorionic gonadotropin (hCG 0.5 IU/ml), forskolin (10 microM), 8-bromo-adenosine-3',5'-cyclic monophosphate (8-Br-cAMP, 1 mM), A23187 (10 microM), phorbol 12-myristate 13-acetate (PMA, 0.01 microM), 25-OH-cholesterol (0.1-10 microM) and pregnenolone (0.1-10 microM) in the presence or absence of ABM-Bu (100 microg/ml). The functions of steroidogenic enzyme including protein expression of the steroidogenic acute regulatory protein (StAR) and cytochrome P450 side-chain cleavage enzyme (P450scc) protein were investigated. Expressions of both P450scc and StAR mRNA have also been explored. We found that ABM decreased progesterone production via an inhibition on (1) the cAMP-PKA and PKC signal transduction pathway, (2) P450scc and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) enzyme activity, (3) P450scc and StAR protein and mRNA expressions and (4) the phosphorylation of ERK1/2 in rat granulosa cells.

Cassava cyanogens and free amino acids in raw and cooked leaves.

Cassava leaves (Manihot esculenta Crantz) constitute the main daily source of protein as supplement to the major staple food, the processed cassava roots in remote rural areas of Africa. Konzo, an upper motoneurone disease with permanent spastic paralysis of both legs, has been reported among populations consuming this unbalanced diet. In commercial pounded cassava leaves residual cyanogens and the presence of inherent potentially toxic non-protein amino acids were analysed to check their safety. The initial total cyanogens before cooking ranged from 35.9+/-0.4 to 107.5+/-0.8 mg HCN (hydrogen cyanide) equivalent kg(-1) dry weight. After cooking, the residual cyanogens were significantly reduced (P<0.05) ranging from 0.30+/-0.04 to 1.9+/-0.2 mg HCN equivalent kg(-1) dry weight, and were below the recommended FAO/WHO safe limit set at 10 mg HCN equivalent kg(-1) dry weight. The total free amino acids and trigonelline (N-methyl-nicotinic acid) detected varied from 10.8 g kg(-1) dry weight to 38.2 g kg(-1) dry weight in the raw and from 7.4 g kg(-1) dry weight to 25.6 g kg(-1) dry weight in the cooked cassava leaves. The non-protein amino acids gamma-amino butyric acid (GABA) and alpha-amino butyric acid (alpha-ABA) were detected. No known potentially toxic non-protein amino acid was found. In konzo-affected areas, cassava leaves with inadequate preparation and cooking can be a non-negligible source of dietary exposure to cyanogens apart from the cassava roots that are suggested to be involved in the aetiology of konzo.

Isocupressic acid blocks progesterone production from bovine luteal cells.

The needles of ponderosa pine (Pinus ponderosa Laws.) were reported to induce abortions when fed to late-term pregnant beef cows in North America. An in vivo study of pregnant cows suggested that isocupressic acid (IA) was the main abortifacient isolated from needles and bark of the pine. However, the mechanism of abortifacient activity of IA is not clear yet. In a pregnant cow, the corpus luteum of the ovary helps the maintenance of pregnancy by its progesterone production. This study involved the IA extracted from the root of the Taiwan cypress (Juniperus formosana) and used a frozen-thawed bovine luteal cell culture system to investigate the action of IA on progesterone production. Thawed bovine luteal cells (1 x 10(5) cells/ml/well) in M199 medium were cultured in 24-well culture plates at 37 degrees C in a 5% CO2 incubator. Ten ml of tested drugs, IA at 1 to 1000 ng/ml and/or ovine luteinizing hormone (oLH) at 1 to 100 ng/microl or 8-bromo-cyclic adenosine monophosphate (8-Br-cAMP) with 0.1-10 mM, were added into each well. After 4 hours of incubation, the media were harvested and assayed for progesterone by an enzyme immunoassay. Progesterone production from cells was the indicator used to evaluate the action of IA. All tested doses of IA significantly inhibited progesterone production in both basal and oLH stimulating conditions. Also those dosages inhibited cyclic adenosine-3',5'- monophosphate (cAMP) stimulation, suggesting a post-cAMP mechanism is involved in the IA action. We concluded that IA can induce pregnant cows to abort partly through blocking luteal function and may be identified as a new abortifacient chemical.

Quinone-type podocarpanes from the bark of Taiwania cryptomerioides.

Three quinone-type podocarpanes, 3beta-hydroxy-13-methoxy-8,12-podocarpadiene-11,14-dione (1), 18-hydroxy-13-methoxy-8,12-podocarpadiene-11,14-dione (2), and 13-methoxy-8,12-podocarpadiene-2,11,14-trione (3) were isolated from the bark of Taiwania cryptomerioides. Their structures were elucidated using spectral methods.

Taxane diterpenoids from the stem bark of Taxus mairei.

Three new 11(15-->1)-abeo-taxanes, taxumairols U-W (1-3), have been isolated from extracts of the stem bark of Formosan Taxus mairei. The structures of 1-3 were identified as 5alpha,7beta,9alpha,13alpha,20-pentaacetoxy-2alpha,10beta,15-trihydroxy-11(15-->1)-abeo-taxene, 5alpha,7beta,9alpha,20-tetraacetoxy-2alpha,10beta,13alpha,15-tetrahydroxy-11(15-->1)-abeo-taxene, and 2alpha,4alpha,7beta,10beta-tetraacetoxy-5beta,20-epoxy-9alpha,13alpha,15-trihydroxy-11(15-->1)-abeo-taxene, respectively, on the basis of 2D NMR techniques including COSY, HSQC, HMBC, and NOESY experiments as well as chemical reactions of compounds 1-3 to give 4 (5alpha,7beta,9alpha,10beta,13alpha,20-hexaacetoxy-2alpha,15-dihydroxy-11(15-->1)-abeo-taxene) and 5 (4alpha,7beta,10beta-triacetoxy-9alpha,13alpha-dibenzoxy-5beta,20-epoxy-2alpha,15-dihydroxy-11(15-->1)-abeo-taxene), which are also novel taxane derivatives. Taxumairols U (1) and V (2) exhibited significant cytotoxicities against human hepatoma tumor cells, while taxumairol W (3) was inactive.

New cyclopropyl-triterpenoids from the aerial roots of Ficus microcarpa.

Four new cyclopropyl-triterpenes, 27-nor-3beta-hydroxy-25-oxocycloartane (1), (22E)-25,26,27-trinor-3beta-hydroxycycloart-22-en-24-al (2), 3beta-acetoxy-15alpha-hydroxy-13,27-cyclours-11-ene (3), 3beta-acetoxy-12alpha-formyloxy-13,27-cycloursan-11alpha-ol (4), together with (23E)-27-nor-3beta-hydroxycycloart-23-en-25-one (5) were isolated from the aerial roots of Ficus microcarpa. Compounds 3 and 4 are rare 13,27-cycloursane-type triterpenes. Their structures were elucidated by spectroscopic and chemical methods.

Allelopathic prenylflavanones from the fallen leaves of Macaranga tanarius.

Two new prenylflavanones, tanariflavanones A (1), and B (2), and one known compound, (-)-nymphaeol-C (3), were isolated from the fallen leaves of Macaranga tanarius. Compounds 1 and 2 showed inhibition of radical growth of lettuce seedlings at 200 ppm. Their structures were elucidated primarily by NMR, circular dichroism, and mass spectroscopic methods.

Nonprotein amino acids in edible lentil and garden pea seedlings.

Commercial edible seedlings of garden pea (Pisum sativum L.) and lentil (Lens culinaris L.) contain high concentration of nonprotein amino acids and trigonelline. Both seedlings grown in the laboratory or purchased in a supermarket were studied by HPLC. Samples from both origins contained trigonelline, alpha-aminoadipic acid, homoserine, beta-(isoxazolin-5-on-2-yl)-alanine (BIA), and gamma-glutamyl-BIA. Garden pea seedlings also contained a uracil-alanine derivative (isowillardiine) in substantial amount. Some of these compounds such as BIA and alpha-aminoadipic acid have neurotoxic activity.

Antagonism of free-radical-induced damage of adlay seed and its antiproliferative effect in human histolytic lymphoma U937 monocytic cells.

The goal of our current research was to investigate the antioxidative effects of methanolic extracts from different parts of adlay seed and their antiproliferative activity in malignant human cells. The methanolic extracts from different parts of adlay seeds were from the hull (AHM), testa (ATM), bran (ABM), and polished adlay (PAM). AHM exhibited greater capacity to scavenge superoxide anion radicals in the PMS-NADH system than ATM, ABM, or PAM. The scavenging capacities of AHM and ATM on hydrogen peroxides were about 20% at a dose of 250 microg/mL. Using the method of deoxyribose degradation to assess damage caused by hydroxyl radicals, AHM was found to inhibit damage in deoxyribose at a higher concentration. However, ATM, ABM, and PAM exhibited prooxidative activity at the same concentration. The inhibitory effect on enzymatic oxidation of xanthine to uric acid was found to follow the order AHM > ATM =. ABM. However, PAM was inactive. All test samples were positive for inhibition of TPA-induced free radical formation on neutrophil-like leukocytes and were found to follow the order AHM > ATM > ABM > PAM. When human histolytic lymphoma U937 monocytic cells were exposed to tert-butyl hydroperoxide, AHM protected the cells against the cytotoxicity caused by tert-butyl hydroperoxide. In addition, AHM exhibited antiproliferative activity against human histolytic lymphoma U937 monocytic cells in a dose-dependent manner. The antiproliferative properties of AHM appear to be attributable to its induction of apoptotic cell death as determined by flow cytometry. These results show that AHM displays multiple antioxidant effects and induces apoptosis of malignant human cells.

Inducible nitric oxide synthase inhibitors from Saposhnikovia divaricata and Panax quinquefolium.

A series of polyacetylenes, falcarinone, panaxynol, falcarindiol, panaxydol, and panaxytriol, were isolated from Saposhnikovia divaricata (Turcz.) Schischk and Panax quinquefolium L. These polyacetylenes were identified as active principles on the inhibition of nitrite production by inducible nitric oxide synthase (iNOS). Treatment with 10 microM of panaxynol, falcarindiol, panaxydol and panaxytriol decreased the LPS/IFN-gamma-stimulated accumulation of nitrite by 71.92 +/- 3.07, 69.95 +/- 3.68, 45.48 +/- 6.11 and 36.85 +/- 8.80%, respectively. The IC50 value of falcarinone, panaxynol, falcarindiol, panaxydol and panaxytriol was > 20, 2.23, 1.98, 6.58 and 9.85 microM, respectively.

Six new compounds from the heartwood of Diospyros maritima.

Six new compounds, diospyrolide (1), diospyrolidone (2), diethyl (2R)-malate (3), 3-(E)-coumaroylbetulin-28-yl ethyl nonanedioate (4), 3-(E)-coumaroylbetulin-28-yl ethyl succinate (5), and 3-(E)-coumaroylbetulin-28-yl ethyl (2R)-2-hydroxysuccinate (6), have been isolated from the heartwood of Diospyros maritima. Compounds 1 and 2 are novel trinorlupanes, and 4, 5 and 6 are lupane derivatives. Their structures were determined using spectral and chemical methods.

Podocarpane-type trinorditerpenes from the bark of Taiwania cryptomerioides.

Five new podocarpane-type trinorditerpenes were isolated from the bark of Taiwania cryptomerioides. Structures of 1beta, 14-dihydroxy-13-methoxy-6,8,11,13-podocarpatetraene (1), 1beta, 14-dihydroxy-13-methoxy-8,11,13-podocarpatriene (2), 13-hydroxy-12-methoxy-8,11,13-podocarpatrien-3-one (3), 14-hydroxy-13-methoxy-1,8,11,13-podocarpatetraen-3-one (4), and 13-methoxy-8,12-podocarpatriene-11,14-dione (5) were determined by NMR, MS, and other spectral and chemical evidence.

New taxanes with an opened oxetane ring from the roots of Taxus mairei.

Two novel taxoids, taxumairols N (1) and O (2), have been isolated from extracts of the roots of Taxus mairei. The structures of 1 and 2 were identified as 7beta,9alpha,10beta,13alpha-tetraacetoxy-2alp ha, 4alpha,5alpha,20-tetrahydroxytax-11-ene and 7beta,9alpha,10beta, 13alpha-tetraacetoxy-1beta,2alpha,4alpha,5alp ha, 20-pentahydroxytax-11-ene on the basis of 1D and 2D NMR techniques including COSY, HMQC, HMBC, and NOESY experiments.

Six new ursane- and oleanane-type triterpenes from the aerial roots of Ficus microcarpa.

Four new ursane-type triterpenes, 3beta-acetoxy-11alpha-methoxy-12-ursene (1), 3beta-acetoxy-11alpha-ethoxy-12-ursene (2), 3beta-acetoxy-11alpha-hydroperoxy-12-ursene (3), 3beta-hydroxy-11alpha-hydroperoxy-12-ursene (4), and two new oleanane-type triterpenes, 3beta-acetoxy-11alpha-ethoxy-12-oleanene (5), 3beta-acetoxy-111alpha-hydroperoxy-12-oleanene (6), together with 3beta-acetoxy-11alpha-hydroxy-12-ursene (7), 3beta,11alpha-diacetoxy-12-ursene (8), 3beta-acetoxy-11alpha-hydroxy-12-oleanene (9), were isolated from the aerial roots of Ficus microcarpa L. f. Their structures were elucidated by spectroscopic and chemical methods.

Six podocarpane-type trinorditerpenes from the bark of Taiwania cryptomerioides.

Six podocarpane-type trinorditerpenes were isolated from the bark of Taiwania cryptomerioides. Their structures, 14-hydroxy-13-methoxy-8,11,13-podocarpatrien-7-one (1), 13-hydroxy-12-methoxy-8,11,13-podocarpatriene (2), 12-hydroxy-13-methoxy-8,11,13-podocarpatriene (3), 14-hydroxy-13-methoxy-8,11,13-podocarpatriene (4), 13-hydroxy-8,11,13-podocarpatriene (5), and 13,14-dihydroxy-8,11,13-podocarpatrien-7-one (6), were determined principally from spectral evidence.

Free amino acids present in commercially available seedlings sold for human consumption. A potential hazard for consumers.

The importance of fresh seedlings for human consumption on European markets continues to increase. Although the contents of free amino acids and potentially toxic free nonprotein amino acids in these fresh and supposedly healthy seedlings is very different from those of the seeds, the crude composition is never mentioned on commercial packages. A commercial product containing seven different kinds of fresh seedlings including kamut, adzuki bean, chickpea, mungbean, pinto bean, garden pea, and lentil has been analyzed by HPLC. Per 100 g of fresh product, 548.2 mg of total free amino acids was found, of which 56.7 mg is free nonprotein amino acids including beta-(isoxazolin-5-on-2-yl)alanine, homoserine, and isowillardiine and the plant hormone trigonelline (N-methylnicotinic acid). The highest amounts of free nonprotein amino acids and trigonelline are found in garden pea (28.3 mg/100 g), mungbean (9.59 mg/100 g), and lentil (7.50 mg/100 g) seedlings. Trigonelline is present in all legume seedlings examined.

A novel cytotoxic C-methylated biflavone from the stem of Cephalotaxus wilsoniana.

Bioassay-directed fractionation of an ethanolic extract of Cephalotaxus wilsoniana has resulted in the isolation of a novel C-methylated biflavone, taiwanhomoflavone-A (1). Its structure was elucidated on the basis of spectroscopic analysis. Taiwanhomoflavone-A is cytotoxic with ED50 values of 3.4, 1.0, 2.0 and 2.5 microg/ml, respectively, against KB epidermoid carcinoma of nasopharynx, COLO-205 colon carcinoma, Hepa-3B hepatoma, and Hela cervix tumor cells.

Dihydrophenanthrenes from Spiranthes sinensis.

Six novel dihydrophenanthrene derivatives, sinensols A-F (1-6), were isolated from the aerial parts of Spiranthesis sinensis. Their structures were determined on the basis of various spectroscopic data, in particular those yielded by MS and 2D NMR techniques.

Cytotoxicity of extractives from Taiwania cryptomerioides heartwood.

The cytotoxicity of the dominant lignans and sesquiterpenoids from Taiwania (Taiwania cryptomerioides Hayata) was investigated. Three human tumor cells including A-549 lung carcinoma. MCF-7 breast adenocarcinoma and HT-29 colon adenocarcinoma were selected to illustrate the structure-cytotoxicity relationships of Taiwania's dominant compounds. Taiwanin A, taiwanin E and dimethylmatairesinol exhibited significant cytotoxicity against three human tumor cells. Among them, taiwanin A possesses the strongest cytotoxic activity. In addition, the morphology-based evaluation, flow cytometric analysis, and DNA fragmentation assays demonstrated that the tumor cell death induced by taiwanin A was due to apoptosis.