• Title/Summary/Keyword: non-aqueous fraction

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Identification of Benzidine Metabolites in Rats by Gas Chromatography/Mass Selective Detector and its Toxicity in vitro (Gas-Chromatography/Mass Selective Detector를 사용하여 쥐의 뇨시료 중 benzidine 대사체의 확인 및 in vitro 독성)

  • 류재천;권오승
    • YAKHAK HOEJI
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    • v.44 no.5
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    • pp.384-390
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    • 2000
  • Metabolism study of the dye, benzidine, was performed by gas chromatography-mass selective detector (GC/MSD) in the urine of rats orally administered 100 mg/kg benzidine. Urine samples were collected in metabolic cages for 0-24, 24-48, and 48-72 hrs. Ten ml of the urine was extracted with XAD-2 resin and the XAD-2 column was eluted with methanol. After evaporation, benzidine and its metabolites were extracted with diethyl ether (for non-conjugated fraction). For conjugated metabolites, $\beta$-glucu-ronidase was added to the aqueous layer that was incubated for 1 hr at 5$0^{\circ}C$ and the aqueous layer was extracted as in non-conjugated fraction. Aliquot of trimethylsilylated derivatives was applied to the GC/MSD. The mutagenicity of benzidine and its acetylated metabolites was tested by histidine/reversion assay. Five metabolites observed and confirmed either by electron impact and chemical ionization modes of the GC/MSD, or authentic compounds were monoacetyl-, diacetyl-, hydroxyacetyl-, hydroxydiacetyl-, and hydroxy-benzidine. Monoacetyl-benzidine was more potent than benzidine in histidine/reversion assay. This data indicates that monoacetylation of benzidine may be one of the metabolites produced in metabolic activation process.

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The Metabolism of (2-$^{14}C$) Mevalonic Acid on Photoperiodic Induction in Grafted Solanum Andigena

  • Bae, Moo;Mercer, E.I.
    • Nuclear Engineering and Technology
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    • v.2 no.2
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    • pp.73-84
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    • 1970
  • The metabolism of sterol precurosor in leaves of Salanum andigena grafted between photoinduced and noninduced plant was investigated with the use of (2-$^{14}$ C) mevalonic acid. By the technique of the preparative gas-liquid chromatography, radioactive compounds of squalene, 4,4’-dimethylsterols and 4-demethylsterol were isolated and determined quantitatively. When labeled mevalonic acid n as applied to leaves radioactivity was extensively incorporated into non-saponifiable materials of lipid fraction and aqueous fraction (ethanol-water fraction). Radioactivity of 14C derived from (2-$^{14}$ C) mevalonic acid was transmissible from photoinduced plant to non-induced plant across the graft union, as tuberization hormone was, and incorporated into the sterols of the non-induced plant. Inhibitors of sterol biosynthesis, SK & F 7997 and nicotinic acid, are effective suppressors of tuber growing, if applied to leaves during photoinduction period. The experimental results suggest that certain substance containing isoprene unit, or sterol-like compound may participate in tuber growing.

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Fate of C-14 Iabelled carbofuran in paddy plants and soil (담수토양(湛水土壤)에 표면처리(表面處理)한 C-14표식(標識) carbofuran의 수도체(水稻體) 및 토양(土壤)에서의 거취(去就))

  • Park, Chang-Kyu;Oh, Sae-Ryang
    • Korean Journal of Environmental Agriculture
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    • v.5 no.2
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    • pp.85-94
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    • 1986
  • To study fate of carbofuran in paddy system, C-14 labelled carbofuran was applied to paddy water containing rice seedlings and time course study was made on the distribution, metabolism and chemical transformation of the systemic insecticide. Carbofuran was readily absorbed by plant root and translocated to shoots where most of the radioactivities were confined to leaf tips. The fact that gradual increases in radioactivities of both aqueous phase extracts and non-extractable fractions of plants (shoots and root) increased with incubation is taken as an evidence that reactions (phase I and II) proceed in rice plants. Carbofuran and its five metabolites were all detected by TLC in organic phase extracts of paddy plants or soil. Evidence was put forward that carbofuran and its five metabolites were all identified as aglycones of conjugates. 7-benzofuranol and 3-hydroxycarbofuran were the most abundant aglycones. Soil microbes appears to have little effects on the metabolism of carbofuran. They increased radioactivity of non-extractable fraction and reduced that of organic phase extracts of paddy soil.

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Enzyme-assisted extraction of Ecklonia cava fermented with Lactobacillus brevis and isolation of an anti-inflammatory polysaccharide

  • Lee, Won-Woo;Ahn, Ginnae;Wijesinghe, W.A.J.P.;Yang, Xiudong;Ko, Chang-Ik;Kang, Min-Cheol;Lee, Bae-Jin;Jeon, You-Jin
    • ALGAE
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    • v.26 no.4
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    • pp.343-350
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    • 2011
  • Fermentation and enzyme-assisted extraction (EAE) improve nutritional and functional properties of foods by increasing the extraction of active compounds, ingestion rates, and body absorption. In this study, we investigated whether applying the EAE process improves the extraction and isolation efficiency of a polysaccharide from fermented Ecklonia cava (FE), which inhibited NO production in lipopolysaccharide (LPS)-activated RAW 264.7 cells. The results showed that the FE using the fungi Candida utilis and two different bacteria, namely Lactobacillus brevis and Saccharomyces cerevisiae increased protein and carbohydrate contents in comparison with those in non-fermented E. cava (NE). Aqueous extracts of fermented E. cava increased extraction yields and carbohydrate content, compared with the aqueous extract of NE. In addition, treating LPS-stimulated RAW 264.7 cells with aqueous extracts resulted in reduced NO production compared to that in LPS-treated cells. Ten EAEs of L. brevis-fermented E. cava (LFE) improved NO inhibitory effects in LPS-activated RAW 264.7 cells and the Viscozyme extract (VLFE) from the resulting extracts showed the highest NO inhibitory effect. We found that the >30 kDa fraction of VLFE led to markedly high inhibition of LPS-induced NO production as compared to that in the <30 kDa fraction. The crude polysaccharide isolated from >30 kDa fraction (VLFEP) consisted of fucose and markedly decreased NO production induced by LPS stimulation. VLFEP could be useful as an anti-inflammatory agent to suppress macrophage activation.

Antiangiogenic activity of non-aqueous fraction from Sparassis crispa extract in human umbilical vein endothelial cells (혈관내피세포에서 꽃송이버섯(Sparassis crispa) 소수성 추출물의 항혈관신생 활성)

  • Han, Jang Mi;Gong, So Youn;Sohng, Jae Kyung;Kang, Yue Jai;Jung, Hye Jin
    • Korean Journal of Food Science and Technology
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    • v.51 no.2
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    • pp.141-146
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    • 2019
  • Sparassis crispa is an edible mushroom that is distributed in Korea, Japan, Europe, and North America. It exerts various biological activities such as immunopotentiation, anti-diabetic, anti-cancer, and anti-inflammatory effects. Recently, we separated the health functional non-aqueous fraction from the chloroform extract of S. crispa (SCF4). In this study, we evaluated the antiangiogenic activity of SCF4 in human umbilical vein endothelial cells (HUVECs). SCF4 effectively inhibited vascular endothelial growth factor (VEGF)-induced cell growth at concentrations ($5-25{\mu}g/mL$) showing no cytotoxic effects. SCF4 inhibited VEGF-induced invasiveness and tube formation ability, which are in vitro angiogenic features of HUVECs, in a dose-dependent manner. In addition, SCF4 markedly suppressed in vivo angiogenesis of chorioallantoic membrane from growing chick embryos without cytotoxicity. Furthermore, SCF4 downregulated the phosphorylation of VEGFR2, AKT, and ERK1/2, which are major angiogenic signal mediators. These results suggest that SCF4 inhibited angiogenesis by suppressing the VEGFR2 signaling pathways without cytotoxicity.

In vitro Anti-malarial Activity of Juniperus Chinensis Extract (향나무 추출물의 항말라리아 효과)

  • Lee, Keyong-Ho;Kim, Byeong-Soo;Ho, Choe-Yeong;Rhee, Ki-Hyeong
    • Korean Journal of Pharmacognosy
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    • v.43 no.3
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    • pp.239-242
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    • 2012
  • This study was carried out to investigate the anti-malarial activity of Juniperus chinensis by in vitro and in vivo system using Plasmodium falciparum chloroquine-sensitive(3D7) and P. falciparum chloroquine-resistant(S20) strains. According to cytotoxicty test on NIH 3T3 cell, the ethanol extract(EtOH), ethylacetate(EtOAc) fraction and aqueous fraction possessed significant anti-malarial activity against both 3D7 and S20 strains at non-toxic concentrations(<100 /). In vitro assay, EtOAc fraction showed notable activity against 3D7 and S20 strains of P. falciparum with $IC_{50}$ values of $37{\pm}2{\mu}g/ml$ and $36{\pm}6{\mu}g/ml$. In animal test using P. falciparum infected human erythrocytes, the treatment of EtOAc fraction significantly inhibited parasitaemia in mice in a dose-dependent manner that is parasitaemia of 42%, 34% and 31% in doses of 10 mg/kg, 20 mg/kg and 40 mg/kg, respectively. The study provides data to support the medicinal importance of the J. chinensis.

Anti-carcinogenic effects of non-polar components containing licochalcone A in roasted licorice root

  • Park, So Young;Kim, Eun Ji;Choi, Hyun Ju;Seon, Mi Ra;Lim, Soon Sung;Kang, Young-Hee;Choi, Myung-Sook;Lee, Ki Won;Yoon Park, Jung Han
    • Nutrition Research and Practice
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    • v.8 no.3
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    • pp.257-266
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    • 2014
  • BACKGROUND/OBJECTIVE: Licorice has been shown to possess cancer chemopreventive effects. However, glycyrrhizin, a major component in licorice, was found to interfere with steroid metabolism and cause edema and hypertension. The roasting process of licorice modifies the chemical composition and converts glycyrrhizin to glycyrrhetinic acid. The purpose of this study was to examine the anti-carcinogenic effects of the ethanol extract of roasted licorice (EERL) and to identify the active compound in EERL. MATERIALS/METHODS: Ethanol and aqueous extracts of roasted and un-roasted licorice were prepared. The active fraction was separated from the methylene chloride (MC)-soluble fraction of EERL and the structure of the purified compound was determined by nuclear magnetic resonance spectroscopy. The anti-carcinogenic effects of licorice extracts and licochalcone A was evaluated using a MTT assay, Western blot, flow cytometry, and two-stage skin carcinogenesis model. RESULTS: EERL was determined to be more potent and efficacious than the ethanol extract of un-roasted licorice in inhibiting the growth of DU145 and MLL prostate cancer cells, as well as HT-29 colon cancer cells. The aqueous extracts of un-roasted and roasted licorice showed minimal effects on cell growth. EERL potently inhibited growth of MCF-7 and MDA-MB-231 breast, B16-F10 melanoma, and A375 and A2058 skin cancer cells, whereas EERL slightly stimulated the growth of normal IEC-6 intestinal epithelial cells and CCD118SK fibroblasts. The MC-soluble fraction was more efficacious than EERL in inhibiting DU145 cell growth. Licochalcone A was isolated from the MC fraction and identified as the active compound of EERL. Both EERL and licochalcone A induced apoptosis of DU145 cells. EERL potently inhibited chemically-induced skin papilloma formation in mice. CONCLUSIONS: Non-polar compounds in EERL exert potent anti-carcinogenic effects, and that roasted rather than un-roasted licorice should be favored as a cancer preventive agent, whether being used as an additive to food or medicine preparations.

Effects of Lycii Fructus on Primary Cultured Chicken Brain Cells

  • Park, Mi-Jung;Chu, Eun-Hye;Lee, Heun-Pa;Kim, Young-Choong
    • Archives of Pharmacal Research
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    • v.14 no.4
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    • pp.325-329
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    • 1991
  • Effects of Lycii Fructus on primary cultured chicken embryonic brain cells were studied by microscopic observation, determination of the activity of pyruvate dehydrogenase complex (PDHC), and syntheses of protein, RNA and DNA. The brain cells were prepared from the brains or 10-day-old chicken embryos and cultured with a deficient medium. The activity of PDHC in the brain cells cultured with a deficient medium was increased to 1.8 times by the addition of $30\;{\mu}g/ml$ of the total methanol extract of Lycii Fructus. To seek the active fraction, total methanol extract was further fractionated by the polarity. The survival rate of neuronal cells was significantly increased by the addition of $100\;{\mu}g/ml$ of the buthanol or aqueous fraction. At this concentration, the significant increase of the syntheses of protein and RNA, but not of DNA, indicates that the fractions may act on the neuronal cells which are known to be non-dividing cells.

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Inhibitory Effect of Hizikia fusiformis Solvent-Partitioned Fractions on Invasion and MMP Activity of HT1080 Human Fibrosarcoma Cells

  • Lee, Seul-Gi;Karadeniz, Fatih;Oh, Jung Hwan;Yu, Ga Hyun;Kong, Chang-Suk
    • Preventive Nutrition and Food Science
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    • v.22 no.3
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    • pp.184-190
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    • 2017
  • Matrix metalloproteinases (MMPs) are endopeptidases that take significant roles in extracellular matrix degradation and therefore linked to several complications such as metastasis of cancer progression, oxidative stress, and hepatic fibrosis. Hizikia fusiformis, a brown algae, was reported to possess bioactivities, including but not limited to, antiviral, antimicrobial, and anti-inflammatory partly due to bioactive polysaccharide contents. In this study, the potential of H. fusiformis against cancer cell invasion was evaluated through the MMP inhibitory effect in HT1080 fibrosarcoma cells in vitro. H. fusiformis crude extract was fractionated with organic solvents, $H_2O$, n-BuOH, 85% aqueous MeOH, and n-hexane (n-Hex). The non-toxicity of the fractions was confirmed by MTT assay. All fractions inhibited the enzymatic activities of MMP-2 and MMP-9 according to the gelatin zymography assay. Cell migration was also significantly inhibited by the n-Hex fraction. In addition, both gene and protein expressions of MMP-2 and -9, and tissue inhibitor of MMPs (TIMPs) were evaluated by reverse transcription-polymerase chain reaction and Western blotting, respectively. The fractions suppressed the mRNA and protein levels of MMP-2, MMP-9 while elevating the TIMP-1 and TIMP-2, with the $H_2O$ fraction being the least effective while n-Hex fraction the most. Collectively, the n-Hex fraction from brown algae H. fusiformis could be a potential inhibitor of MMPs, suggesting the presence of various derivatives of polysaccharides in high amounts.

A Study on Char Characteristics of Fire Retardant Treated Douglas Fir (난연처리된 Douglas Fir의 탄화특성에 관한 연구)

  • Park Hyung-Ju;Oh Kyu-Hyung;Kim Eung-Sik;Kim Hong
    • Fire Science and Engineering
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    • v.19 no.2 s.58
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    • pp.105-110
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    • 2005
  • We tested the char characteristics of fire retardant treated Douglas fir at each of five constant external irradiance levels $(10,\;15,\;20,\;25\;및\;35kW/m^2)$. A Cone heater was used to expose the wood specimens to the heat flux. The size of specimens is 100- by 100- by 50-mm and the kinds of specimens are non-treated wood(N) and treated wood(F2 and f4) by water soluble fire retardants. The water-soluble fire retardants were made from mixture of aqueous solutions of monoammonium phosphate, sodium borate and zinc borate, and those are used for immersion of Douglas fir. In result of test, char fraction of fire retardant treated Douglas fir showed a considerably low char fraction than it of non-treated wood irrespective of increase of external heat flux. And char fractions has low levels with increase of fire retardant content. Burning rate of non-treated wood(N) was showed a relatively high burning rate than it of fire retardant treated wood(F2 and F4). And difference of burning rate shown more rapidly in high external irradiance than low external irradiance. When the external heat flux is $35kW/m^2$, average char rate of non-treated wood is rapidly about twice than fire retardant treated wood. Water-soluble fire retardants mixed in this study find out it has fire suppression and adiabatic effect by char layer from results of char fraction, burning rate, and char depth and rate.