• Title/Summary/Keyword: [$^{14}C$]-acetate

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Detection of Antiinflammatory Agents from Natural Products as Inhibitors of Cyclooxygenase I and II

  • Lee, Dong-Hee;Kang, Sam-Sik;Chang, Il-Moo;Mar, Woong-Chon
    • Natural Product Sciences
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    • v.3 no.1
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    • pp.19-28
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    • 1997
  • Constitutive cyclooxygenase (COX-I) is present in cells under physiological conditions, whereas inducible cyclooxygenase (COX-II) is induced by some cytokines, mitogens, and endotoxin presumably in pathological conditions such as inflammation. We have evaluated the inhibitory effects of solvent fractionated extracts of natural products on the activities of COX-I and COX-II. Oxygen uptake COX assay was performed, as a primary screening from the tissue extracts of bovine seminal vesicles (BSV), by monitoring the initial rate of oxygen uptake using an oxygen electrode. Additionally, we evaluated plant extracts for the inhibitory effects of COX-I (in HEL cells) and COX-II (in lipopolysaccharide activated J774A.1 macrophages) using thin layer chromatography of prostanoids produced from $^{14}C-labelled$ arachidonic acid (AA). The use of such models of COX-I and COX-II assay will lead to the identification of specific inhibitors of cyclooxygenases with presumably less side effects than present therapies. Inhibitory effects of 50 kinds of plant extracts on the COX-I and COX-II activities were determined and the active fractions were found in the ethyl acetate fractions of Dryopteris crassirhizoma (roots), Amomum cardamomum (roots), Triticum aestivum (seeds), Perilla sikokiana (leaves), Anemarrhena asphodeloides (roots). Especially, the ethyl acetate fraction of Dryopteris crassirhizoma (roots), which exhibited the strong inhibition against BSV COX $(IC_{50},\;65.4\;{\mu}g/ml)$, COX-I $(IC_{50},\;8.5\;{\mu}g/ml)$, and COX-II $(IC_{50},\;17.2\;{\mu}g/ml)$, is under investigation to isolate active principles using activity-guided fractionation method.

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Effects of Dietary Nitrogen Sources on Fiber Digestion and Ruminal Fluid Characteristics in Sheep Fed Wheat Straw

  • Tan, Z.-L.;Lu, D.-X.;Hu, M.;Niu, W.-Y.;Han, C.-Y.;Ren, X.-P.;Na, R.;Lin, S.-L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.10
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    • pp.1374-1382
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    • 2001
  • Fifteen Inner Mongolian wethers with permanent ruminal and duodenal cannulas were used to study the effects of dietary rumen-undegradable protein (RUP) to rumen-degradable protein (RDP) ratios or protein sources on fiber digestion in the gastrointestinal tract and ruminal fluid characteristics. Fiber digestion and ruminal fermentation were not affected (p>0.05) by dietary RUP to RDP ratios (from 1.54 to 0.72). Soybean meal supplementation improved ruminal digestion. Fish meal supplementation increased (p<0.05) the ruminal degradability of fiber. The different RUP to RDP ratios (from 1.54 to 0.72) did not influence (p>0.05) ruminal fluid pH, but there were differences (p<0.05) in ruminal fluid $NH_3-N$ concentration because of urea replacement. Soybean meal as a dietary protein source decreased (p<0.05) ruminal fluid pH and increased (p<0.05 or p<0.01) $NH_3-N$, acetate, propionate and butyrate concentrations in the rumen. Fish meal as a dietary protein source decreased (p<0.05 or p<0.01) ruminal $NH_3-N$ and acetate concentrations and increased (p<0.05) ruminal propionate concentration. It can be concluded that dietary protein sources have more significant effect on fiber digestion and ruminal fermentation than different dietary RUP to RDP ratios, when the dietary crude protein requirements of growing sheep are satisfied.

Partial Characterization of Allelopathic Substances in Sorghum Stem by Different Organic Solvents and pH (수수 줄기에 함유(含有)된 타감물질(他感物質)의 용매(溶媒)와 pH에 따른 특성구명(特性究明))

  • Kim, S.Y.;De Datta, S.K.;Robles, R.P.;Kim, K.U.;Lee, S.C.;Shin, D.H.
    • Korean Journal of Weed Science
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    • v.14 no.1
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    • pp.42-48
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    • 1994
  • The toxic compounds of sorghum stem extracts were isolated by different organic solvents and pH, and characterized and quantified in terms of their inhibition of seed germination and seedling growth in Echinochloa colona(L.) Link and radish(Raphanus sativus L.). Sequential partitioning of stem extract with various organic solvents with increasing polarity showed that all fractions of hexane, ethyl ether, methylene chloride, ethyl acetate, and the aqueous remainder inhibited germination and seedling growth in E. colona. Of the five fractions, the ethyl ether fraction had the greatest inhibitory effect on E. colona. Further separation of the ethyl ether fraction at different pH(pH 2-11) showed that phytotoxic compounds were acidic. The result indicates that the phytotoxin present in the stem extract may be nonpolar and acidic.

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Extraction and Purification of EGCG(Epigallocatechin Gallate) from Green Tea (녹차로부터 EGCG(Epigallocatechin Gallate)의 추출 및 정제)

  • Gang, Ji-Hun;Park, Yeong-Gwang;Jeong, Seong-Taek;No, Gyeong-Ho
    • KSBB Journal
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    • v.14 no.5
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    • pp.517-522
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    • 1999
  • A green tea used in this experiment was cultivated at Bosung (Chonnam) and purchased from a domestic market. The extract at 5$0^{\circ}C$ water from the powder of green tea partitioned with chloroform and ethyl acetate. The resulting solution was further purified with a chromatographic column (4.6$\times$250 mm, 15${\mu}{\textrm}{m}$, Lichrospher 100RP-18). Finally separation was achieved on a $\mu$-Bondapak $C_18$(3.9$\times$300mm, 10${\mu}{\textrm}{m}$) column. The elution order of the catechin compounds contained in the green tea was EGC(Epigallocatechin, C(catechin), EC(Epicatechin), EGCG(Epigallocatechin Gallate) and ECG(Epicatechin Gallate). From the experimental results the mobile phase for isolating EGCG from the extract consisted of 0.1% acetic acid in water/acetonitrile, 87/13%(v/v). The flow rate of mobile phase was 1.0 $m\ell$/min, and UV wavelength was fixed at 280 nm. 121.3 mg of EGCG, higher than 98% of purity, was obtained from 5 g of dry green tea.

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Chemical Composition and Antitumor Apoptogenic Activity of Methylene Chloride Extracts from the Leaves of Zanthoxylum schinifolium (Zanthoxylum schinifolium잎의 methylene chloride 추출물의 화학적 조성 및 암세포에 대한 세포자살 유도활성과 그 작용기전)

  • Kim Jun-Seok;Jun Do-Youn;Woo Mi-Hee;Rhee In-Koo;Kim Young-Ho
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.546-554
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    • 2006
  • To understand antitumor activity of Zanthoxylum schinfolium, which has been used as an aromatic and medicinal plant in Korea, the cytotoxic effect of various organic solvent extracts of its leaves on human tumor cells were investigated. Among these extracts such as methanol extract (SL-13), methylene chloride extract (SL-14), ethyl acetate extract (SL-15), n-butanol extract (SL-16), and residual fraction (SL-17), SL-14 appeared to contain the most cytotoxic activity against leukemia and breast cancer cells tested. The methylene chloride extra.1 (SL-14) possessed an apoptogenic activity causing apoptotic DNA fragmentation of human acute leukemia Jurkat T cells via mitochondrial cytochrome c release into cytoplasm, subsequent activation of caspase-9 and caspase-3, and cleavage of PARP, which could be negatively regulated by antiapoptotic protein Bcl-xL. The GC-MS analysis of SL-14 revealed that the twenty-two ingredients of SL-14 were 9,19-cyclolanost-24-en-3-ol (15.1%), 2-a-methyl-17, b-hop-21-ene (15.1%), 15-methyl-2,3-dihydro-1H benzazepin (11.95%), phytol (10.38%), lupeol (9.92%), 12-methylbenzofuran (8.23%), hexadecanoic acid (5.96%), cis,cis,cis-9,12,15-octadecatrienoic acid-methyl-ester (5.49%), 9,12,15-octadecatrienoic acid-methylester (3.59%), 15-methyl-4-(1-methylethylidene)-2-(4-nitrophenyl) (3.36%), hexadecanoic acid methyl ester (1.93%), vitamine E (1.88%), beta-amyrin (0.96%), and auraptene (0.89%). These results demonstrate that the cytotoxicity of the methylene chloride extract of the leaves of Z. schinifolium toward Jurkat T cells is mainly attributable to apoptosis mediated by mitochondria-dependent caspase cascade regulated by Bcl-xL, and provide an insight into the mechanism underlying antitumor activity of the edible plant Z. schinifolium.

STUDIES OF ACNE TREATMENT USING ORIENTAL HERBS(New Approach to select anti-acne agents)

  • Nam, Chun-Ja;Han, Y.G.;Kim, S.J.;Kim, J.H.;Oh, J.Y.;Park, J.W.;Lee, H.
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.25 no.4 s.34
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    • pp.111-121
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    • 1999
  • Acne vulgaris, the most common skin disease. can be formed as only a few comedons or severe inflammatory lesions. The pathogenesis of acne involves various factors; excessive androgen, excessive sebum production, abnormal alteration of follicular epithelium, proliferation of Propionibacterium acnes, and inflammation. We investigated acne therapy using oriental herbs described in the Korean traditional medical book(Dong-ui-bo-gam). Oriental herbs(Angelica daurica, Arctium lappa, Coptidis rhizoma, and Glycyrrhiza glabra) were chosen based on their respective property of sebum control, anti-inflammatory activity, and anti-bacterial activity. We examined the effect of acne treatment, in terms of chemotactic inhibition, lipogenesis inhibition, and anti-bacterial activity for P. acnes. 1. Neutrophil chemotaxis assay; P. acnes secrete chemotactic factors and other pro-inflammatory extracellular products. Neutrophil chemotactic activity of P. acnes was measured by 48-well chemotaxis method. Angelica daurica clearly suppressed chemotactic activity of P. acnes. 2. Using sebaceous gland of hamster ear lipogenesis assay; Sebaceous lipogenesis was measured using ear biopsies by incubation or $C^{14}$-acetate in culture media. The $C^{14}$-labeled lipids were extracted and determined by liquid scintilation counting. Coptidis rhizoma markedly inhibited sebum production. 3. Anti-bacterial assay for P. acnes(MIC test); Glycyrrhiza glabra showed anti-bacterial activity. P. acnes did not develop resistance against Glycyrrhiza glabra. Retinoids are effectively to inhibit sebum production and regulate follicular keratinization process, with little anti-inflammatory activity. Angelica daurica suppressed neutrophil chemotaxis, Coptidis rhizoma inhibited sebum production, and Glycyrrhiza glabra showed anti-bacterial activity against P. acnes. A combined formulation of Angelica daurica, Coptidis rhizoma. and Glycyrrhiza glabra is expected to provide effective acne treatment.

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Evaluation of Bioactivity of Ti-6Al-7Nb Alloys with Various Hydrothermal Treatment Times (열수처리 시간에 따른 Ti-6Al-7Nh 합금의 생체활성 평가)

  • Kwon O. S.;Choi S. K.;Park K. B.;Lee M. H.;Bae T. S.;Lee O. Y.
    • Korean Journal of Materials Research
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    • v.14 no.12
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    • pp.876-884
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    • 2004
  • This study was to investigate whether the bioactivity of the anodized and hydrothermally treated Ti-6Al-7Nb alloy were affected by the time of hydrothermal treatment. Anodizing was performed at current density 30 $mA/cm^2$ up to 300 V in electrolyte solutions containing $DL-{\alpha}-glycerophosphate$ disodium salt hydrate $(DL-{\alpha}-GP)$ and calcium acetate (CA). Hydrothermal treatment was done at $300^{\circ}C$ for 30 min, 1 hr, 2 hrs, and 4 hrs to produce a thin film layer of hydroxyapatite (HA). The bioactivity was evaluated from HA formation on the surfaces in a Hanks' solution with pH 7.4 at $36.5^{\circ}C$ for 10, 20, and 30 days. Anodic oxide films were porous with pore size of $1\sim4{\mu}m\;and\;3\sim4{\mu}m$ thickness. The anodic oxide films composed with strong anatase peak with presence of rutile peak, and showed the increase in intensity of anatase peak after hydrothermal treatment. It was shown that the intensity of anatase peak increased with increasing the time of hydrothermal treatment but was no difference in rutile peak. The corrosion voltage was the highest in the group of hydrothermal treatment for 2 hrs (Ecorr: -338.6 mV). The bioactivity in Hank's solution was accelerated with increasing the time of hydrothermal treatment.

A Study on the Extraction of Thermostable Pectinesterase from Valencia Orange (Valencia 오렌지로부터 내열성 Pectinesterase의 추출)

  • Hou, Won-Nyoung;Walker, Brigdet L.
    • Korean Journal of Food Science and Technology
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    • v.27 no.5
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    • pp.658-665
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    • 1995
  • Low yield of a thermostable pectinesterase(TSPE) from citrus fruits has made its detailed study extremely tedious and difficult; therefore, maximizing TSPE extraction is desirable. It is assumed that TSPE is bound to the cell components via ionic linkage and covalent bonds. Therefore, in this study, variations in extraction time, pH, NaCl concentration and commercial enzyme preparations were used to increase the yield of TSPE from Valencia orange. The largest recovery of TSPE, obtained by heating extracted pectinesterase(PE) at $70^{\circ}C\;for\;5{\sim}10$ minute, was achieved using actate buffer(pH 4.14) with 1 M NaCl and 0.2% $Cytolase^{TM}$ 104(a mixture of cellulase, hemicellulase and pectinase; Genecor, Inc). The two aquous phase partitioning with 5.0% Triton X-114 could be used as a tool for separation of thermolabile pectinesterase(TLPE) and TSPE from crude PE. Also, water extraction and $0{\sim}0.3$ ammonium sulfate fractionation could be used for removing non-pectinesterase protein.

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STUDIES OF ACNE TREATMENT USING ORIENTAL HERBS (New Approach to select anti-acne agents)

  • Chunja Nam;Y.G. Han;Kim, S.J.;Kim, J.H.;Oh, J.Y.
    • Proceedings of the SCSK Conference
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    • 1999.10a
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    • pp.111-121
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    • 1999
  • Acne vulgaris, the most common skin disease. can be formed as only a few comedons or severe inflammatory lesions. The pathogenesis of acne involves various factors; excessive androgen, excessive sebum production, abnormal alteration of follicular epithelium, proliferation of Propionibacterium acnes, and inflammation. We investigated acne therapy using oriental herbs described in the Korean traditional medical book (Dong-ui-bo-gam). Oriental herbs (Angleica daurica. Arctium lappa. Coptidis rhizoma, and glycyrrhiza glabra) were chosen based on their respective property of sebum control, anti-inflammatory activity, and anti-bacterial activity. We examined the effect of acne treatment, in terms of chemotactic inhibition, lipogenesis inhibition, and anti-bacterial activity for P. acnes. 1. Neutrophil chemotaxis assay ; P acnes secrete chemotactic factors and other pro-inflammatory extracellular products. Neutrophil chemotactic activity of P. acnes was measured by 48-well chemotaxis method. Angelica daurica clearly suppressed chemotactic activity of P. acens. 2. Using sebaceous gland of hamster ear lipogenesis assay; Sebaceous lipogenesis was measured using ear biopsies by incubation of $C^{14}$ -acetate in culture media. The $C^{14}$ -labeled lipids were extracted and determined by liquid scintilation counting, Coptidis rhizoma markedly inhibited sebum production, 3. Anti-bacterial assay for P. acnes (MIC test) Glycyrrhiza glabra showed anti-bacterial activity. P. acnes did not develop resistance against Glycyrrhiza glabra. Retinoids are effectively to inhibit sebum production and regulate follicular keratinization process, with little anti-inflammatory activity. Angelica daurica suppressed neutrophil chemotaxis. Coptidis rhizoma inhibited sebum production, and Glycyrrhiza glabra showed anti-bacterial activity against P. acnes. A combined formulation of Angelica daurica. Coptidisr hizoma and Glycyrrhiza glabra is expected to provide effective acne treatment.ent.ive acne treatment.

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Analysis of $\triangle^9$-Tetrahydrocannabinol and 11-nor-9-carboxytetrahydrocannabinol in Hair by Gas Chromatography/Mass Spectrometry (가스크로마토그라피/질량분석기에 의한 모발중 대마성분 분석)

  • 양원경;한은영;박용훈;임미애;정희선
    • YAKHAK HOEJI
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    • v.48 no.3
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    • pp.207-212
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    • 2004
  • An analytic method was developed for the quantitation of $\Delta$$^{9}-$ tetrahydrocannabinol (THC) and 11-nor-9-carboxy THC (THC-COOH) in human hair. After hair samples were pulverized using Freezer Mill, deuterated internal standards were added and digested in 1 N NaOH at $100^{\circ}C$ water bath for 30 min. Digest solutions were extracted by 5 ml hexane:ethyl acetate (90:10) after acidification with acetic acid. The organic phase was evaporated under N 2 and derivatized by BSTFA (with 1% TMCS) at $85^{\circ}C$ for 45 min. The derivatized solution was separated on HP-5MS column ($30m{\times}0.25mm{\times}0.25mm$) and detected using EI-GC-MS with selective ion monitoring mode. The assay of calibration was ranged from 5 to 100 ng/50 mg hair ($r^2$>0.99) for THC and THC-COOH. Within and between-run precision were calculated at 6, 30, 60 ng/50 mg hair with coefficients of variation less than 11%. Within and between run accuracies at the same concentrations were$\pm$14% and $\pm$30% of target for both analytes, respectively. Absolute and relative recovery at 10 and 100 ng were 60∼91%. The method was used to detect and quantify THC and THC-COOH in cannabis abuser's hairs (N = 16) and SRM (N=5, THC 1 ng/mg, NIST). We detected THC and THC-COOH in only one hair sample. In SRM, % accuracy was 93% (range 86∼103%) and precision (% CV) was 8.14. We began to set up a quantitative analysis of THC and THC-COOH using EI-GC-MS. Continuously, we need to modify and develop this method in order to apply for identification in cannanbis users' hair.