A stable isotopic approach for quantifying aquaculture-derived organic matter deposition dynamics in the sediment of a coastal fish farm.

The stable carbon and nitrogen isotopic compositions (δ13C and δ15N) of sedimentary organic matter (OM) in fish farms (FFs) were investigated to quantify the aquaculture-derived OM deposition dynamics in the sediment. The dual isotopic compositions of mixed OMs in surface sediments at FFs differed significantly (p < 0.05) from those at reference sites, indicating an increased deposition of fish feces or uneaten feed in the sediments. Furthermore, OM source apportionments revealed that the quantitative contribution of fish feces (23.3 mg g-1 dw) during farming activities was significantly higher than that of other natural OM sources (C3 plants and phytoplankton). After the disassembly of fish cages, the deposited fish feces may be preferentially degraded through processes that require a large amount of oxygen consumption (<0.1 kg C m-2 yr-1). Our isotopic approach may be helpful for assessing the impact of FF wastes and for taking measures to minimize environmental deterioration.

Spatial variability of water quality and sedimentary organic matter during winter season in coastal aquaculture zone of Korea.

We investigated the composition, source, and reactivity of sedimentary organic matter (OM) present in aquaculture systems along the Korean coast. A multi-isotopic approach was applied to 216 surface sediments from five sections: western (W)-1, W-2, southern (S)-1, S-2, and eastern (E)-1 sections. The 15N-enriched total nitrogen signatures in the surface sediments of the W-1 section may indicate that a substantial fraction of sedimentary OM has anthropogenic origins exported through the Han River. Simultaneously, the deposition of allochthonous OM is predominant in the S sections (Jinhae and Masan Bays). The 34S-depleted patterns at the S-2 section may indicate that sulfate reduction occurring at the sedimentary boundary contributes to active OM decomposition, depending on the increased sedimentation of the aquaculture-derived OM. Our results highlight that isotopic-based source tracking near aquaculture systems provide important information for identifying anthropogenic contamination in coastal marine sediments and for improving environmental management.

Formononetin prevents ovariectomy-induced bone loss in rats.

The major risk factor of postmenopausal osteoporosis is estrogen deficiency. Hormone replacement therapy is efficacious against osteoporosis, but it induces several significant adverse effects. In this study, therefore, we compared therapeutic potencies of three phytoestrogens: genistein, daidzein, and formononetin. Our result showed that in Saos-2 cells, formononetin and genistein (5 x 10(-7) M) treatment increased alkaline phosphatase activity by 33.0 +/- 5.8% and 21.1 +/- 4.0%. Genistein inhibited osteoclast formation in a dose-dependent manner. In OVX rats, formononetin-treated groups given 1 and 10 mg/kg/day displayed increased trabecular bone areas (TBAs) within the tibia. Genistein- and daidzein-treated groups also displayed increased tibial TBAs. TBAs of the lumbar vertebrae were higher in all treated groups than in the control group. In conclusion, formononetin as well as other isoflavones, such as daidzein and genistein, inhibited bone loss caused by estrogen-deficiency.

Rat growth-hormone release stimulators from fenugreek seeds.

Bioassay-guided fractionation of MeOH extract from fenugreek (Trigonella foenum-graecum L.) seeds resulted in the isolation of two rat growth-hormone release stimulators in vitro, fenugreek saponin I (1) and dioscin (9), along with two new, i.e., 2 and 3, and five known analogues, i.e., 4-8. The structures of the new steroidal saponins, fenugreek saponins I, II, and III (1-3, resp.), were determined as gitogenin 3-O-beta-D-xylopyranosyl-(1-->6)-beta-D-glucopyranoside, sarsasapogenin 3-O-beta-D-xylopyranosyl-(1-->6)-beta-D-glucopyranoside, and gitogenin 3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranoside, respectively. Fenugreek saponin I (1) and dioscin (9) caused ca. 12.5- and 17.7-fold stimulation of release, respectively, of rat growth hormone from rat pituitary cells, whereas gitogenin (5) showed moderate activity. To our knowledge, this is the first study to demonstrate that steroidal saponins stimulate rat growth-hormone release in rat pituitary cells.

Triterpenoid saponins from the seeds of Pharbitis nil.

From the seeds of Pharbitis nil (Convolvulaceae), two new oleanene-type triterpene glycosides, pharbitosides A (1) and B (2), together with beta-sitosterol, beta-sitosterol glucoside (daucosterol), caffeic acid, and methyl caffeate were isolated. The structure of pharbitoside A (1) was elucidated to be queretaroic acid 3-O-alpha-L-rhamnopyranosyl-(1-->2)-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranoside (1). Pharbitoside B (2) is a 21alpha-hydroxyoleanolic acid saponin carrying the same sugar moiety as that of pharbitoside A (1).

Induction of growth hormone release by glycyrrhizae radix on rat.

Induction of growth hormone (GH) by Glycyrrhizae Radix (GR), one of the most popular herbal medicine, and its major ingredients were studied in rat pituitary cells in vitro and in vivo assay. The MeOH extract and the n-hexane (HX) fraction of GR induced rat GH (rGH) release up to 1.89 times (0.34 +/- 0.04 nM) and 4.59 times (0.83 +/- 0.03 nM), compared to the basal level (p < 0.05). Among many ingredients isolated and purified from GR both glycyrrhetinic acid and glycyrrhizin induced significantly rGH release compared to the control (p < 0.05). After an intravenous injection of rat growth hormone releasing hormone (rGHRH) (10 microg/kg) as positive control, in SD rats, Tmax of plasma rGH level was 10 min, C(max) was 3.84 +/- 0.01 nM (n = 3), and enhanced plasma rGH level returned to the baseline in 90 min. Both AUC(0-90) (area under the curve) of plasma rGH level after HX fraction and that after rGHRH administration were increased significantly from the basal level, respectively (p < 0.01). In conclusions, HX fraction is the most active fraction of MeOH extract of GR in rGH induction.

Induction of growth hormone release by dioscin from Dioscorea batatas DECNE.

In this study, dioscin was isolated from Dioscoreae Rhizoma (DR), which is the rhizome of Dioscorea batatas D(ECNE). that inhabits broad areas of Korea and Japan. To determine whether dioscin induced growth hormone (GH) release, we evaluated its induction effects on GH release both in vitro and in vivo. The 70% methanol extract of DR, and its n-hexane and n-BuOH fractions, induced rat GH (rGH) release in rat pituitary cells 10-fold, 8-fold, and 5- fold higher than the control (0.36 +/- 0.02 nM), respectively (p < 0.05 each). The dioscin-induced rGH release of the cells was concentration-dependent and its ED(50) was 1.14 x 10(-5) M. Within 90 minutes after intravenous administration of 10 microg/kg (p < 0.05 at t(max)), dioscin caused the greatest increase in rGH concentration (C(max)) in the rat plasma (34.16 +/- 14.10 ng/ml) (n = 4), which was twice as high as the control group (12.88 +/- 3.29 ng/ml) (n = 27).

Norditerpenoid and dianthramide glucoside alkaloids from cultivated Aconitum species from korea.

A new norditerpene alkaloid named 8-O-methylhypaconine (1) was isolated along with twelve known alkaloids from the underground parts of an unknown species of Aconitum plant culti vated in Korea. Among the known alkaloids, two dianthramide glucosides, N-(2'-beta-glucopyra nosyl-5'-hydroxysalicyl)-5-hydroxyanthranilic acid methyl ester (2) and N-(2'-beta-glucopyranosyl-5'-hydroxysalicyl)-5-hydroxy-6-methoxyanthranilic acid methyl ester (3), were isolated from Aconitum plants for the first time. The structures were established on the basis of chemical and spectroscopic methods.

High performance liquid chromatographic analysis of isoflavones in medicinal herbs.

Phytoestrogens have been used as a food supplement to prevent osteoporosis. The isoflavones in the phytoestrogens are daidzein, genistein and formononetin which are present in various herbs. This study examined the quantity of isoflavones in medicinal herbs, which can be used as a phytoestrogen supplement; soybean. These isoflavones were quantified using high performance liquid chromatography (HPLC) with a UV/VIS detector. The concentration of daidzein in Puerariae Radix was 10,436.16 +/- 2,143.83 mg/kg of the dried herb, which was much higher than that extracted from soybeans, 341.47 +/- 18.96 mg/kg. The amount of genistein in Sophorae flavescentis Radix (336.09 +/- 50.89 mg/kg) was approximately 11 times higher than that extracted from soybean (30.03 +/- 7.17 mg/kg). The level of formononetin in Dalbergiae odoriferae Lignum, 2,189.14 +/- 136.46 mg/kg, was the highest among the herbs tested. The total isoflavone content of Puerariae Radix was approximately 30 times higher than that extracted from soybean. Therefore, plants from the family Leguminosae, particularly Puerariae Radix, can be a good source of phytoestrogens.

Herbal extract prevents bone loss in ovariectomized rats.

This research aims to test a new drug candidate based on a traditional medicinal herb, F1, an herbal extract obtained from Astragalus membranaceus and its main ingredient, 1-monolinolein that may have fewer side effects and less uterine hypertrophy. In vitro experiments, human osteoblast-like cell lines, MG-63 and Saos-2, were analyzed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and an alkaline phosphatase (ALP) assays. Mouse osteoclasts were induced through a calcium-deficient diet and inhibition effects were measured. In vivo experiments were done using ovariectomized (OVX) rats for 9 weeks. At necropsy, uterus weights were measured, trabecular bone area (TBA) of tibia and lumbar vertebra were measured bone histomorphology. In results, cell proliferation and ALP activity in Saos-2 by ether F1 or 1-monolinolein did not increased significantly compared to the control. The F1 inhibited osteoclast development (IC25 = 3.37 x 10(-5) mg/mL) less than 17beta-estradiol. The OVX rats administered F1 (2 mg/kg/day and 10 mg/kg/day) showed an increase in TBA of the tibia significantly (136.3 +/- 4.2% and 138.5 +/- 10.3% of control). In conclusions, the herbal extract, F1 inhibited tibia and lumbar bone loss and did not cause uterine hypertrophy. However, 1-monolinolein, the main ingredient of the herbal extract, did not inhibit bone loss.

Effects of Eucommiae Cortex on osteoblast-like cell proliferation and osteoclast inhibition.

Methanol extract (MeOH), n-hexane (Hx), chloroform (CHCl3), ethyl acetate (EA), butanol (BuOH) and aqueous (H2O) fractions of Eucommiae Cortex including geniposidic acid (GA), geniposide (GP) and aucubin (AU) were tested for their therapeutic efficacy on osteoporosis. The contents of GA, GP and AU in the cortex and leaf of Eucommia ulmoides Oliver were quantified by HPLC. The effect of Eucommiae Cortex on the induction of growth hormone (GH) release was studied by using rat pituitary cells. The proliferation of osteoblast-like cells increased by herbal extracts was assayed using a tetrazolium (MTT), alkaline phosphatase (ALP) activity, and [3H]-proline incorporation assays. The inhibition of osteoclast was studied by using the coculture of mouse bone marrow cells and ST-2 cells. As a result, the GA, GP and AU were present in the cortex more than in the leaf of E. ulmoides Oliver. The MeOH (1 mg/mL), Hx, CHCl3 and EA fractions (each 20 microg/mL) had potent induction of GH release. The CHCl3 exhibited the potent proliferation of osteoblasts. The AU, GP and GA were increased proliferation of osteoblasts. In addition, GA (IC50: 4.43 x 10(-7) M), AU and GP were significantly inhibited proliferation of osteoclast. In summary, it is thought that the components in a part of the fractions of Eucommiae Cortex participate in each step of mechanism for activating osteoblast to facilitate osteogenesis, and suppress osteoclast activity to inhibit osteolysis.

Study of substance changes in flowers of Pueraria thunbergiana Benth. during storage.

Puerariae Flos is a traditional herbal medicine that has long been used as a treatment for colds, diabetes, and hangovers. The herbal medicine contains a wide variety of isoflavones such as kakkalide, tectoridin, and tectorigenin. This study demonstrates that the substances undergo a certain degree of change depending on the storage period by the method of HPLC and 13C-NMR, and that the HPLC analysis can be used to determine the freshness of Puerariae Flos.

Induction of growth hormone by the roots of Astragalus membranaceus in pituitary cell culture.

The traditional Asian medicinal herb, roots of Astragalus (A.) membranaceus (Leguminosae), is used for many purposes, some of which are purported to stimulate the release of growth hormone in vivo. Extracts of A. membranaceus were tested to determine whether they stimulate the release of growth hormone in rat pituitary cell culture. A. membranaceus was extracted sequentially with 80% ethanol (fraction A), n-hexane (fraction B); the test compound from the herbal extraction was isolated using silica gel column chromatography and was identified with spectral data. Test compound was also extracted by traditional boiling water methods. Induction of growth hormone in pituitary cell culture was conducted with isolated compounds and extracted fractions of A. Radix (dried roots of A. membranaceus). The fraction A was not active in the rat pituitary cell culture, but the fraction B derived from the ethanol fraction stimulated the release of growth hormone in culture. Six compounds from fraction B (1-6) were isolated and identified previously. The compounds 1,2-benzendicarboxylic acid diisononylester (1), beta-sitosterol (2), and 3-O-beta-D-galactopyranosyl-beta-sitosterol (5) did not induce growth hormone release in the culture. Formononetin (3), 9Z,12Z-octadecadienoic acid (4), stigmast-4-en-6beta-ol-3-one (6) and 98-E, a mixture of 1'-9,12-octadecadienoic acid (Z,Z)-2',3'-dihydroxy-propylester (7) and 1'-hexadecanoic acid-2',3'-dihydroxy-propylester (8) stimulated the release of growth hormone in the rat pituitary cell culture significantly compared to the control. In conclusions, four compounds isolated from extracts of A. Radix induced growth hormone release in the rat pituitary cell culture. The 98-E isolate was the most active inducer of growth hormone release.

Pharmacokinetic analysis of rhein in Rheum undulatum L.

This research aims to identify the main active compounds of Rhei undulati Rhizoma (roots of Rheum undulatum LINNE) and determine the types of anthraquinones absorbed into the body and their pharmacokinetic parameters. The boiling-water extract of the herb was administered to 12 healthy volunteers (9 men/3 women) at a dosage of 100 mg/kg the anthraquinone levels in plasma were determined with TLC, HPLC, and LC-MS. Rhein was the only anthraquinone compound absorbed by the body as determined for plasma analysis of the volunteers. The elimination rate constant of rhein in Rhei undulati Rhizoma was 0.23+/-0.02/h and the half life was 3.38+/-0.35 h. This experiment confirmed that rhein is the most important active compound absorbed by the body among anthraquinones contained in Rhei undulati Rhizoma, indicating that rhein is a promising marker substance to evaluate Rhei Rhizoma and Rhei undulati Rhizoma.