• Title/Summary/Keyword: Dose fractionation

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Albumin Fractions from Different Species Stimulate In Vitro Progesterone Production by Granulosa Cells in Buffalo

  • Taneja, R.;Bansal, P.;Sharma, M.K.;Singh, D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.11
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    • pp.1559-1563
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    • 2002
  • The ovarian follicular fluid was found to contain steroidogenesis stimulatory protein similar to albumin from human and buffalo. Therefore, the albumins from various species, commercial and purified, were studied for their steroidogenic effect on progesterone secretion by granulosa cells from buffalo ovaries, during culture. A dose of $20{\mu}g$ of bovine serum albumin was optimum to exhibit maximum progesterone secretion on day 6 of culture, in medium ($350{\mu}l$) containing $10^5$ cells. Among commercial albumins, chicken albumin showed highest effect on progesterone secretion, which was followed by albumins from goat, bovine, human, sheep and rat, respectively at day 6 of culture. The albumins were also purified from blood serum of buffalo, goat and rat using salt fractionation, ion-exchange chromatography, gel filtration and SDS-PAGE. The highest stimulatory effect on progesterone secretion was shown by albumin purified from buffalo blood serum and lowest by that from rat blood. Comparatively the buffalo and goat albumins were more biologically active than commercial albumins. The presence of some active molecules conjugated with freshly purified albumins may be responsible for better stimulatory effect.

Anti-oxidative Activities of 3'-Hydroxygenkwanin from the Flower Buds of Daphne genkwa in Caenorhabditis elegans

  • Park, Sung-Hoon;Cui, Xun;Ahn, Dalrae;Lee, Eun Byeol;Cha, Dong Seok;Jeon, Hoon;Zee, Ok Pyo;Kim, Youn-Chul;Kim, Dae Keun
    • Natural Product Sciences
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    • v.20 no.2
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    • pp.80-85
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    • 2014
  • In the course of screening for antioxidant compounds from natural plants in Korea by measuring the radical scavenging effect, a methanol extract of the flower buds of Daphne genkwa S. et Z. (Thymelaeaceae) was found to show a potent antioxidant activity. Subsequent activity-guided fractionation of methanol extract of D. genkwa led to the isolation of four compounds from the ethyl acetate soluble fraction. The chemical structures were elucidated as genkwanin (1), 3'-hydroxygenkwanin (2), apigenin (3), and tiliroside (4) by spectroscopic techniques. Among them, compound 2 showed the significant anti-oxidative effect on DPPH. And compound 2 showed the significant riboflavin-and xanthine-originated superoxide quenching activities. To verify the antioxidant enzymatic activities of compound 2, the SOD enzymatic activity was measured spectrophtometrically using prepared Caenorhabditis elegans homogenates. The results showed that compound 2 was able to elevate SOD activity of C. elegans in a dose dependent manner. Moreover, compound 2 decreased the intracellular ROS accumulation of worms.

Antioxidant Activity of a Red Seaweed Polysiphonia morrowii Extract

  • Je, Jae-Young;Ahn, Chang-Bum;Oh, Myung-Joo;Kang, So-Young
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.124-129
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    • 2009
  • Antioxidant activities of the extract of red seaweed, Polysiphonia morrowii, were evaluated using several in vitro assay systems. Activity-guided fractionation revealed that the 90% MeOH fraction of the P. morrowii extract exhibited the highest antioxidant activity, and that this fraction had a high total phenolic content ($135.7{\pm}5.0\;mg$ gallic acid/g extract). Therefore, the antioxidant activities of the 90% MeOH fraction against 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radical, reducing power, ferrous chelating, and hydrogen peroxide were investigated. The results revealed that the antioxidant activities of the 90% MeOH fraction were similar and/or superior to that of commercial antioxidants such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT). In addition, the ability of the 90% MeOH fraction to inhibit oxidative damage to DNA was assessed by measuring the conversion of the supercoiled pBR322 plasmid DNA to the open circular form. The 90% MeOH fraction was found to significantly protect this hydroxyl radical-induced DNA damage in a dose-dependent manner. Taken together, these findings suggest that the 90% MeOH fraction of P. morrowii extract and/or its constituents has the potential for use as a new bioresource of antioxidants.

Isolation and Characterization of Intestinal Immune System Modulating and Anticancer Active Fractions from the Herbal Prescriptions

  • Hwang, Jong-Hyun;Jeong, Jae-Hyun;Yu, Kwang-Won
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.323-329
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    • 2009
  • The prescriptions (DB-1-DB-5) were prepared with the active herbal medicines, Acanthopanax senticosus, Glycyrrhiza uralensis, Polygonatum odoratum, and Cichorium intybus. The most active crude polysaccharide fraction (DB-2-3), which was isolated through the fractionation of hot-water extract from DB-2, was significantly reduced by periodate oxidation (52.7 and 63.7%) on intestinal immune system modulating and anticancer activity. When DB-2-3 was further fractionated by column chromatographies, DB-2-3IIc-2 showed the most potent activities. In addition, DB-2-3IIc stimulated the proliferation of bone marrow cells via Peyer's patch in dose-dependent pattern by oral administration. The metastasis of colon 26-M3.1 lung carcinoma had significantly inhibited in mice fed DB-2-3IIc at 1 mg/mouse (43.8%). DB-2-3IIc-2 mainly contained uronic acid (46.1%) and 42.5% of neutral sugar with a small amount of protein (7.6%), and component sugar analysis also showed that DB-2-3IIc-2 was composed Ara, Gal, and GalA (molar ratio; 0.50:0.63:1.00). It may be suggested that activities of DB-2-3IIc-2 are resulted from pectic polysaccharides containing a polygalacturonan moiety with side chain of neutral sugars, such as Ara and Gal.

Induction of Apoptosis in Human Oral Epidermoid Carcinoma Cells by Sophoraflavanone G from Sophora flavescens

  • Cha, Jeong-Dan;Jeong, Mi-Ran;Lee, Young-Eun;Lee, Kyung-Yeol
    • Food Science and Biotechnology
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    • v.16 no.4
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    • pp.537-542
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    • 2007
  • Sophora flavescens AITON (Leguminosae) is a typical traditional Korean medical herb considered to exhibit antibacterial, anti-inflammatory, and antipyretic effects, and is also used for the treatment of skin and mucosal ulcers, sores, diarrhea, gastrointestinal hemorrhage, arrhythmia, and eczema. In this study, the compound sophoraflavanone G was isolated from the dried roots of S. flavescens by bioassay-guided fractionation. We then investigated the effects of various concentrations of sophoraflavanone G on cell viability and the induction of apoptosis in KB cells after an incubation of 24 hr. The results were determined by the following methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-terazolium bromide (MTT) assay, Hoechst-33258 dye staining, flow cytometry (cell cycle), and Western blotting for caspase-3 and poly (ADP-ribose) polymerase (PARP). We found sophoraflavanone G induced the apoptosis of KB cells in a dose-dependent manner that was verified by DNA fragmentation, apoptotic bodies, the sub-G1 ratio, caspase-3 activity, and cleavage of PARP. These results suggest that sophoraflavanone G has potent anti-proliferative effects on human oral epidermoid carcinoma cells, with the induction of apoptosis.

Antidiabetic Coumarin and Cyclitol Compounds from Peucedanum japonicum

  • Lee, Sung-Ok;Choi, Sang-Zin;Lee, Jong-Hwa;Chung, Sung-Hyun;Park, Sang-Hyun;Kang, Hee-Chol;Yang, Eun-Young;Cho, Hi-Jae;Lee , Kang-Ro
    • Archives of Pharmacal Research
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    • v.27 no.12
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    • pp.1207-1210
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    • 2004
  • The antidiabetic activity-guided fractionation and isolation of the 80% EtOH extracts from Peucedani Radix (Peucedanum japonicum, Umbelliferae) led to the isolation and characterization of a coumarin and a cyclitol as active principles, that is, peucedanol 7-O-${\beta}$ -D-glucopyranoside (1) and myo-inositol (2). Their structures were identified by spectroscopic methods. Compound 1 showed 39% inhibition of postprandial hyperglycemia at 5.8 mg/kg dose, and compound 2 also significantly inhibited postprandial hyperglycemia by 34% (P<0.05).

Susceptibility of Oral Bacterial to Sophoraflavanone G isolated from the Root of Sophora flavescens

  • Kim, Kyong-Heon;Kim, Baek-Cheol;Yun, Ju-Bong;Jeong, Seung-Il;Kim, Hong-Jun;Ju, Young-Sung
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.17 no.3
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    • pp.33-37
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    • 2004
  • Objective: The aim of this work is to investigate the antibacterial activity of the Sohporaflavanone G isolated from Sophora flavescens (S. flavescens), as the development of microbial resistance to antibiotics make it essential to constantly look for new and active compounds effective against pathogenic bacteria. Method : Sophoraflavanone G was isolated from the dried roots of Sophora flavescens Aiton (Leguminosae) by bioassay?guided fractionation. We investigated the effect of sophoraflavanone G on oral bacterial at various concentrations after incubation of 24 h in strains in the dose?dependent manner. Results: The structure of active compound, Sophoraflavanone G having a lavandulyl group at C?8, was elucidated on the basis of spectral data especially 1H?NMR and I3C?NMR. The antimicrobial activity showed that Sophoraflavanone G exhibited antimicrobial activilies against all the bacteria tested (MICs, 0.39 - 6.25 ㎍/ml). Sophoraflavanone G showed the strong antimicrobial activity against all the facultative bacteria and microaerophilic bacteria (MICs, 0.78 - 1.56 ㎍/ml) and also Sophoraflavanone G showed the strong antimicrobial activity against obligate anaerobic bacteria (MICs, 0.39 - 6.25 ㎍/ml).

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Bioproduction and Anticancer Activity of Biosurfactant Produced by the Dematiaceous Fungus Exophiala dermatitidis SK80

  • Chiewpattanakul, Paramaporn;Phonnok, Sirinet;Durand, Alain;Marie, Emmanuelle;Thanomsub, Benjamas Wongsatayanon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1664-1671
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    • 2010
  • A new biosurfactant producer was isolated from palm-oil-contaminated soil and later identified through morphology and DNA sequencing as the yeast-like fungus Exophiala dermatitidis. Biosurfactant production was catalyzed by vegetable oil, supplemented with a basal medium. The culture conditions that provided the biosurfactant with the highest surface activity were found to be 5% palm oil with 0.08% $NH_4NO_3$, at a pH of 5.3, with shaking at 200 rpm, and a temperature of $30^{\circ}C$ for a 14-day period of incubation. The biosurfactant was purified, in accordance with surfactant properties, by solvent fractionation using silica gel column chromatography. The chemical structure of the strongest surface-active compound was elucidated through the use of NMR and mass spectroscopy, and noted to be monoolein, which then went on to demonstrate antiproliferative activity against cervical cancer (HeLa) and leukemia (U937) cell lines in a dose-dependent manner. Interestingly, no cytotoxicity was observed with normal cells even when high concentrations were used. Cell and DNA morphological changes, in both cancer cell lines, were observed to be cell shrinkage, membrane blebbling, and DNA fragmentation.

Inhibitors of Nitric Oxide Syntheasis from Phellinus pini in Murine Macrophages (낙엽송층버섯의 Nitric Oxide 생성저해 물질)

  • Jang, Hyun-Jin;Kim, Ahn-Keun;Pyo, Myoung-Yun;Yang, Ki-Sook
    • YAKHAK HOEJI
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    • v.51 no.6
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    • pp.430-434
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    • 2007
  • The anti-inflammatory activity of fruit body of Phellinus pini was investigated by activity-guided fractionation. From the screening of each fraction for the inhibitory activity of NO production in lipopolysaccaride (LPS) activated RAW 264.7 cells, methanol extract and its hexane soluble fraction of Phellinus pini exhibited inhibition of NO production compared with LPS control without toxicity. The hexane soluble fraction showed dose-dependent inhibition of NO production. The active hexane fraction was repeatedly chromatographed over silica gel, ergosta-7,24(28)-dien-3-ol(1) and ergosterol peroxide (2) were isolated and identified. Ergosterol derivatives were inhibited NOS activation, $IC_{50}$ of them were $18.9{\pm}3.9{\mu}M$ (1) and $20.4{\pm}4.5{\mu}M$ (2).

Anti-Helicobacter pylori Effect of Costunolide Isolated from the Stem Bark of Mgnolia Sieboldii

  • Park, Jong-Beak;Lee, Chong-Kyo;Park, Hee-Juhn
    • Archives of Pharmacal Research
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    • v.20 no.3
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    • pp.275-279
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    • 1997
  • Helicobacter pylori (H. pylorl) infection is now established as the major pathogenic factor in chronic gastritis and peptic ulcer disease. in addition, there is accumulating evidence that H. pylori plays an important role in the process of gastric carcinogenesis. On the other hand, oriental traditional medicines have been used for stomach disease for thousands of years. In the present study, methanol extract from the stem bark of Magnolia sieboldii (M. sieboldii) and its components were investigated on their inhibitory effects against urease activity and growth of H. pylori in vitro. The methanol extract of M. sieboldii significantly inhibited the growth of H. pylori ATCC 43504 at 5 mg/ml. From the further fractionation, the chloroform fraction inhibited the bacterial growth dose-dependently. Among four fractions separated from the chloroform fraction by silica gel column chromatography, MS-C-2 was the most potent. Costunolide was isolated from the MS-C-2 subtraction by preparative TLC and recrystallization using n-hexane. Anti-H. pylori effect of costunolide was investigated using one commercial strain (H. pylori ATCC 43504) and three clinical strains (H. pylon 4, 43, 82548). Costunolide exhibited potent anti-H. pylori activity, and the MIC was around $100-200{\mu}g/ml$. However, costunolide had no inhibitory effect of H. pylori urease activity at the concentration used for the growth inhibition assay. From these results, we conclude that costunolide inhibits the, growth of H. pylori by the independent manner of H. pylori urease inhibition.

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