• Korean Journal of Plant Resources
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• v.31 no.1
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• pp.16-23
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• 2018

Paeonia japonica is a perennial flowering plant used in traditional medicine therapy. The purpose of this study was to investigate the effect of water extract and solvent fractions obtained from P. japonica on anti-oxidative, anti-thrombin, anti-invasive and pro-apoptotic activities in YD-10B cells, human oral squamous carcinoma cell line. Water fraction revealed the highest extraction yield at 11.44% (w/w). Anti-oxidative activity was the highest in ethyl acetate fraction (85.13%). In the thrombin inhibitory activity test, ethyl fraction was the highest, with a value of 87.54%. Release and activation of MMP-2/pro-MMP-2 ratio in thrombin-treated YD-10B cells were significantly inhibited in the ethyl acetate fraction. At a concentration of $120{\mu}g/m{\ell}$ water extract and solvent fractions of P. japonica inhibited cell proliferation in YD-10B cells except water fraction. Pro-apoptotic effect on human oral squamous carcinoma cell using the Bax/Bcl-2 ratio analysis was higher in water extract than other fractions. These findings suggest that the ethyl acetate fraction of P. japonica potentiates a promising antioxidant, anti-thrombin and anti-invasive agents.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.2
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• pp.617-622
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• 2014

Voacanga globosa (Blanco), a plant endemic to the Philippines, is traditionally used especially by indigenous people of Bataan in the treatment of ulcers, wounds and tumorous growths. This study aimed to provide scientific evidence to therapeutic properties by determining cytotoxic and pro-apoptotic activity of HPLC fractions from leaves on HCT116 human colon carcinoma and A549 human lung carcinoma cell lines. Ethanolic extraction was performed on V globosa leaves followed by hexane and ethyl acetate partitioning. Silica gel column chromatography and high performance liquid chromatography (HPLC) produced MP1, MP2 and MP3 fractions. Cytotoxic activity of the fractions was determined through MTT assay against the cancer cell lines HCT116 and A549 and the non-cancer AA8 Chinese hamster ovarian cell line. Pro-apoptotic activities of the most active fractions were further assessed through DAPI staining, TUNEL assay and JC-1 mitochondrial membrane potential assay with HCT116 cells. While the MPI fraction exerted no significant activity against all cell lines tested, MP2 and MP3 fractions demonstrated high toxicity against HCT116 and A549 cells. The MP3 fraction induced formation of apoptotic bodies, condensed DNA and other morphological changes consistent with apoptosis of HCT116 cells and TUNEL assay showed significant increase in DNA fragmentation over time. In these cells, the MP3 fraction also induced mitochondrial membrane destabilization, which is generally associated with the beginning of apoptosis. Phytochemical analysis demonstrated the presence only of saponins and terpenoids in the MP3 fraction. The results indicate that the MP3 fraction exerts cytotoxic activity on HCT116 cells via induction of apoptosis triggered by loss of mitochondrial membrane potential crucial for cell survival.

• Journal of Food Hygiene and Safety
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• v.2 no.2
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• pp.57-65
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• 1987

ABSTRACT - To examine antihypertensive components of Ganoderma lucidum in Korea, two kinds of fruiting bodies (J and K) were used for extraction with water and the extracts were purified by ethanol precipitation and dialysis. Three fractions, i.e., the aqueous total extract(A), the ethanol supernatant(B} and the purified precipitate(C), were compared for antihypertensive activity in anesthetized spontaneously hypertensive rats (SUR). Although fractions A and B showed the activity, fraction C did not. Particularly, fraction B of sample K produced 44.3% reduction in diastolic blood pressure and 30.6% reduction in heart rate after i.v. administration of a dose of 10 mg/kg. Direct effects of this fraction B to the heart were observed in the isolated blood perfused heart preparation of the dog. It induced positive chronotropic and inotropic responses dose-dependently in the case of sample J. In the case of sample K, marked chronotropic and inotropic effects on atrial muscle but not on ventricular muscle were induced. In both samples, coronary blood flow (CBF) was dose-relatedly increased.reased.

• Journal of Life Science
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• v.8 no.4
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• pp.400-409
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• 1998

Formation of adduct was studied in benzo(a)pyrene(BP)- and doxorubicin(Dx)-treated human NC-37 cells and isolated nuclei. Major adducts formed were determined by fluorescence absorption spectrophotometery and DNA-lin-ked protein assay. When isolated nuclei were exposed to carcinogens BP and DMBA, and anticancer drugs m-AMSA, ellipticine and Dx, varying degrees of adduct formation occured between DNA-protein complex and these drugs. When the mixture was centrifuged 1.7 M sucrose solution, binding BP and DMBA appeared to be similar between the sediment and the supernatant. When the sediment was centrifuged again with 0.35% polymin-P, the amount of BP bound was 2-fold greater in the protein(1077$\pm$55cpm) than in DNA fraction (470$\pm$20cpm), whereas that of DMBA was 1.6-fold greater in the DNA than in protein fraction. In the case of m-AMSA, ellipticine and Dx, the amount of binding was slightly greater in supernatant than in sediment in centrifugation with 1.7 M sucrose, and more than 3 times greater in the DNA- than in protein- fraction in centrifugation with 0.35% polymin P. DNA fractions which associated with a subset of nonhistone chromosomal protein were isolated from NC-37 cells exposed to $^{3}$H-BP and $^{14}$C-Dx. They were separated into two distince components DNA-S and DNA-P by centrifugation with 2M Nacl chromatin extraction. The results indicated that the amount of $^{3}$H-BP bound was 6.0-fold greater in DNA-P as compared with DNA-S, while that of $^{14}$C-Dx binding appreaed to be 6.2-fold greater in DNA-S than in DNA-P fraction. When $^{3}$H-BP binding wasdetermined in the presence of cold Dx, the amount of binding was reduced only in the DNA-P fraction, indicating that the interaction between DNA and protein is decreased. Gene expression by these drugs, BP treated cells were increased to compare with nomal cells but reduced by treatment with BP-Dx. These results suggest that the protein moiety which tightly bound to DNA-P fraction may play an important role in the regulation of gene expression.

• Archives of Pharmacal Research
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• v.26 no.1
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• pp.34-39
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• 2003

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-Ο-$\beta$-D-galactopyranosyl-$\beta$-sitosterol (5) did not induce growth hormone release in the culture. Formononetin (3), 9Z, 12Z-octadecadienoic acid (4), stigmast-4-en-6$\beta$-o1-3-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.

• KSBB Journal
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• v.26 no.2
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• pp.165-171
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• 2011

Fractional solvent extraction by organic solvents such as hexane, chloroform, ethylacetate, and butanol was carried out using 70% ethanol extract of apple flower leaves. Biological activities including antioxidant, whitening, antimicrobial and anti-wrinkle activities were investigated and bio-active materials of the extracts were identified using GC/MSD. Among the tested solvent fractions, ethylacetate fraction showed the highest total polyphenol content (1218.94 ${\mu}g/mL$), and flavonoid (140 ${\mu}g/mL$). The DPPH radical scavenging activities was over 80% at a dry matterbased concentration of 200 ${\mu}g/{\mu}L$ and SOD-like activity was over 90% at 50 ${\mu}g/mL$ concentration in ethylacetate fraction that was slightly lower than of ascorbic aicd. Tyrosinase inhibition activity related to skin-whitening was over 60% by ethylacetate fraction of 100 ${\mu}g/mL$. As an anti-aging effect, elastase inhibitory activity was about 45% in ethylacetate fraction. Also, it showed a significantly antimicrobial activity against P. acenes. From GC/MSD analysis, a characteristic peak of high content in ethylacetate fraction was identified as kaempferol, which has been reported as a bioactive compound.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.112-118
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• 2013

This study was performed to investigate the possible use of Eisenia bicyclis (EB) ethanol extract to inhibit activity against lipase. In tests, the lipase inhibitory activity of EB ethanol extract was noted as being 43, 27, and 24% at concentrations of 5, 2.5, and 1 mg/ml, respectively. Isolation was carried out by liquid and liquid extraction, silica-gel column chromatography, and HPLC. The results showed that the lipase inhibitory activity of the ethyl acetate (EA) fraction from EB ethanol extract exhibited the strongest lipase inhibitory activity with an $IC_{50}$ value of 1.31 mg/ml. The EA fraction was separated using silica-gel column chromatography and we obtained 22 sub-fractions. Amongst them, the EA1 fraction showed the highest lipase inhibitory activity with an $IC_{50}$ value of 0.54 mg/ml. Eight peaks were obtained from the EA1 fraction by HPLC. Fraction 5 also showed a strong lipase inhibitory activity with an $IC_{50}$ value of 0.37 mg/ml. The fraction 5 was identified as dieckol and the inhibition pattern analyzed from Lineweaver-Burk plots revealed a non-competitive inhibitor. These results suggest that EB has potential as a natural anti-obesity agent.

• Applied Biological Chemistry
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• v.45 no.3
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• pp.152-156
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• 2002

Soils near abandoned zinc mines were known to be contaminated with arsenic-rich mining by-products. To examine the potential impacts of arsenic- contaminated soils on plant growth, surface soils were subjected to sequential extraction. Results revealed that 54% and 74% total As and 74% total extractable As were bound to iron hydrous oxide, and water soluble fraction was below detection limit. Arsenic faction extracted using the Koran standard method(dissolution of metals via treatment of 1 N HCI) was strongly correlated with the Fe-bound As fraction ($r^2=0.884**$). Arsenic level in rice plant roots was the highest with a maximum value of 154.9 mg/kg, whereas it was below 0.6 mg/kg in grains. Arsenic level in rice plant roots was strongly correlated with those of Al-bound As ($r^2=0.821**$) and 1N HCI-extractable As levels ($r^2=0.801**$).

• The Korean Journal of Food And Nutrition
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• v.31 no.6
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• pp.775-782
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• 2018

The purpose of this study was to investigate effects of Crataegus pinnatifida var. psilosa extracts obtained from 95% methanol and water. Methylene chloride, ethyl acetate, and methanol were used to fractionate the 95% methanol extract and each fraction was testified total polyphenol contents, electron donating abilities, the scavenging abilities of superoxide anion radical, as well as hydrogen peroxide. Extraction yields of 95% methanol and water from Crataegus pinnatifida var. psilosa were 25.40%, 23.12% respectively. Total polyphenol contents were $28,708.0{\pm}1,755.05{\mu}g\;GAE/mL$ in 95% methanol, revealing the highest among them, $12,726.67{\pm}479.33{\mu}g\;GAE/mL$ in water extract, $15,854.67{\pm}498.38{\mu}g\;GAE/mL$ in methanol fraction, $11,810.67{\pm}584.48{\mu}g\;GAE/mL$ in ethyl acetate fraction, and $5294.67{\pm}190.36{\mu}g\;GAE/mL$ in methylene chloride fraction. Total polyphenol contents revealed significant differences (p<0.05) between the solvents. In the experiment of the electron donating ability, water extract revealed $84.33{\pm}0.1%$ scavenging ability, the highest. Other extracts and fractions were $81.8{\pm}1.11%$ for water, $79.73{\pm}1.32%$ for ethyl acetate fraction, $75.73{\pm}2.17%$ for methylene chloride fraction, and $42.1{\pm}5.01%$ for methanol fraction, the lowest electron donating ability. Electron donating abilities revealed significant difference (p<0.05) between the solvents. In the experiment of superoxide anion radical scavenging ability, ethyl acetate fraction($0.0026{\pm}0.0002$) had the highest scavenging ability, and the others revealed slight increase rather than decrease in scavenging ability. Hydrogen peroxide scavenging ability revealed the highest in methanol fraction ($-0.00206{\pm}0.00165$) and the others were as follow; water extract ($0.00157{\pm}0.00249$), 95.0% methanol extract ($0.005{\pm}0.0036$), methylene chloride fraction ($0.0039{\pm}0.00364$), and ethyl acetate fraction ($0.0002{\pm}0.00059$).

• Ocean and Polar Research
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• v.26 no.2
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• pp.231-243
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• 2004

Sequential extraction was carried out on twenty two subsamples of three ferromanganese crusts from three seamounts (Lemkein, Lomilik, and Litakpooki) near the Marshall Islands in the western Pacific. The extraction was designed to fractionate Fe-Mn crust forming elements into low defined groups: (1) exchangeable and carbornate, (2) Mn-oxide, (3) Fe-oxyhyd.oxide, and (4) residual fraction. X-ray diffraction result shows that target material were well removed by each extraction step except for CFA in phosphatized crusts generation. According to chemical analysis of each leachate, most of elements in the Fe-Mn crusts are bound with two major phases. Mn, Ba, Co, Ni, Zn, (Fe, Sr, Cu, and V) are strongly bounded with Mn-oxide $({\delta}-MnO_2)$ phase, whereas Fe, Ti, Zr, Mo, Pb, Al, Cu,(V, P, and Zn) show chemical affinity with Fe-oxyhydroxide phase. This result indicates that significant amount of Al, Ti, and Zr can not be explained by detrital origin. Ca, Mg, K, and Sr mainly occur as exchangeable elements and/or carbonate phase. Outermost layer 1 and inner layer 2 which are both young crusts generations are similar in chemical speciations of elements. However, some of Fe-oxyhydroxide bounded elements (Pb, Y, Mo, Ba, Al, and V) in phosphatized innermost layer 3 are released during phosphatization and incorporated into phosphate (Pb, Y, Mo, and Ba) or Mn-oxide phase (Al and V). Our sequential extraction results reveal that chemical speciations of elements in the hydrogenetic crusts are more or less different from interelemental relationship calculated by statistical method based on bulk chemistry.