• Title/Summary/Keyword: Activity assay

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Identification of Antioxidative Constituents from Polygonum aviculare using LC-MS Coupled with DPPH Assay

  • Shin, Hyeji;Chung, Hayeon;Park, Byoungduck;Lee, Ki Yong
    • Natural Product Sciences
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    • v.22 no.1
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    • pp.64-69
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    • 2016
  • A method for simultaneously identifying antioxidative compounds was developed using time-based LC-MS coupled with DPPH assay regardless of the time consuming process. The methanolic extract of Polygonum aviculare (Polygonaceae) showed significant DPPH radical scavenging activity. Time-based DPPH assay for simultaneous identification of active compounds from the extracts of P. aviculare was used. Major peaks of ethyl acetate fraction of P. aviculare showed high DPPH radical scavenging activity. A simple phenolic compound (1) and six flavonoids (2-7) were isolated from the ethyl acetate fraction of P. aviculare by silica gel and sephadex LH-20 column chromatography. The structures of seven compounds were determined to be protocatechuic acid (1), catechin (2), myricitrin (3), epicatechin-3-O-gallate (4), avicularin (5), quercitrin (6), and juglanin (7) based on the analysis of the $^1H$-NMR, $^{13}C$-NMR and ESI-MS data. All compounds exhibited significant antioxidant activity on DPPH assay and active compounds were well correlated with predicted one.

UV Spectrometric Assay of Epoxide Hydrolase Activity of Microbial Cell Biocatalysts (자외선분광기를 이용한 미생물 세포 생촉매의 에폭사이드 가수분해효소 활성평가)

  • Kim, Hee Sook;Lee, Eun Yeol
    • Applied Chemistry for Engineering
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    • v.16 no.3
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    • pp.456-459
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    • 2005
  • UV spectrometric assay for measurement of epoxide hydrolase activity was tested for efficient screening of whole cell activity of epoxide hydrolase. Epoxide hydrolase activities were determined by measuring the amount of p-nitrostyrene diol (pNSD), which was the hydrolysis product of p-nitrostyrene oxide (pNSO). Enantioselective hydrolysis of racemic pNSO using epoxide hydrolase activity of Rhodosporidium toruloides was monitored by UV spectrometric assay, and the relevant $K_m$ and $V_m$ for R. toruloides were determined as $2.457nmol/min{\cdot}mg$ and 1.078 mM, respectively.

Antioxidant Effects of Noni (Morinda citrifolia) Extracts Treated with Hel and Trypsin (염산과 트립신으로 처리한 노니(Morinda citrifolia) 추출물의 항산화 효과)

  • Choi, Hye Young;Choi, Byung Chul;Sim, Sang Soo
    • YAKHAK HOEJI
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    • v.49 no.5
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    • pp.410-415
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    • 2005
  • To investigate biological activity of noni extracts treated with HCl and trypsin, we measured the antioxidant activity through vitro assay and cellular system. Both water and lipid soluble fraction of noni extracts dose-dependently scav­enged DPPH radical. Superoxide scavenging activity of lipid soluble fraction after treating HCl and trypsin was significantly more potent than those of other fractions in NBT/xanthine oxidase assay, which suggests that antioxidant activity of noni extracts was increased by the treatment with HCl and trypsin. In antioxidant assay using RBL 2H3 cells, water soluble frac­tion of noni extracts had little effect on silica-induced reactive oxygen species generation, whereas lipid soluble fraction inhibited in a dose dependent manner. In non-treated noni extracts, effect of water soluble fraction on silica/$CuSO_4$-induced lipid peroxidation was more potent than that of lipid soluble fraction. However, the effects of noni extracts were reversed in noni extracts treated with HCl and trypsin. These data suggest that water soluble substances may be converted into lipid soluble substances by the treatment with HCl and trypsin. From the above results, it is suggested that lipid soluble fraction of noni extracts contain antioxidant used in vitro assay and RBL 2H3 cellular system. Such an effect of noni extracts may be increased by the treatment with HCl and trypsin.

Clq-Coated Microtitre Enzyme-linked Immunosorbent Assay for Measuring the Anticomplementary Activity of Intravenous Immunoglobulin Preparations (Clq-coated ELISA법을 이용한 정맥용 면역글로불린제제의 항보체성 측정)

  • 강혜나;김순남;신광훈;허숙진
    • YAKHAK HOEJI
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    • v.45 no.6
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    • pp.656-663
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    • 2001
  • The quality of an intravenous immunoglobulin preparation (IVIG) is reflected by the degree of nonspecific activation of complements, the so-called anticomplementary activity (ACA). ACA of aggregates in IVIG was investigated using method by the European Pharmacopoeia and Clq-coated microtiter enzyme-linked immunosorbent assay (ELISA). Both the EP method and the ELISA method showed a dose response curve with the amount of complements bound increasing with the percentage content of aggregates in immunoglobulin standard. The correlation between the two tests was good (r=0.96, r=0.99). However, the correlation was not found when the ACA (EP method) of IVIG product was compared with its aggregate percentage. These results emphasize that the method of aggregate formation affects ACA and that estimation of the percentage distribution of aggregates by HPLC may not reflect ACA. In analysing WIG product for Clq binding activity test with the ELISA, the result by using Protein A-HRP correlated with aggregate percentage (r=0.84). But the correlation decreased (r=0.48) when the result used Protein A-AP(having poorer sensitivity than HRP) was compared with aggregate percentage. As a result, some variation between the two methods, due to differences in assay principles, is to be expected. However, ELISA technique has the advantage in that it is easier to perform, more precise and less subject to reagent variability, and is the more suitable screening method than HPLC analysis.

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Evaluation of Anti-inflammatory activity of Fissistigma poilanei and Rubus laxus Focke

  • Lee, Soo Jung;Suh, Mee-Hyun;Kim, June-Hyun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.75-75
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    • 2019
  • The aim of the study was to determine the anti-inflammatory activities of the plants with origin of China. The Fissistigma poilanei, which is a species of plant in the family Annonaceae and Rubus laxus Focke, which is a species of plant in the family Rosaceae from China were tested for anti-inflammatory activities. Samples were prepared using 95% ethanol using nitric oxide (NO) assay for assessing the anti-inflammatory activity. NO assay experiment showed that extracts of the Fissistigma poilanei give 28.3% increases in anti-inflammatory activity and extracts of Rubus laxus Focke give 57.1% decreases in anti-inflammatory activity, compared to control. To determine the cell toxicity, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used. MTT assay experiment showed that Fissistigma poilanei and Rubus laxus Focke might have less toxicity of 60.5% and 62.3%, respectively, compared to control. Taken together, these experiments showed that Fissistigma poilanei extracts might have significantly higher anti-inflammatory activities and relatively lower toxicity, compared to control. This implies that this study might give a tool to develop a new potential anti-inflammatory therapeutic candidate.

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Antimicrobial, Antioxidant and Hemolytic Activity of Water-soluble Extract of Mottled Anemone Urticina crassicornis

  • Lee, Ye Jin;Kim, Chan-Hee;Oh, Hye Young;Go, Hye-Jin;Park, Nam Gyu
    • Fisheries and Aquatic Sciences
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    • v.18 no.4
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    • pp.341-347
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    • 2015
  • We evaluated the biological activities of five water extracts of tissue of the mottled anemone Urticina crassicornis. Most extracts exhibited broad-spectrum antimicrobial activity as determined by ultrasensitive radial diffusion assay (URDA) against gram-positive and -negative bacteria, including a fish pathogen, Aeromonas hydrophila, but no activity against fungi. The activity of the extracts was abolished by tryptic digestion, indicating that protein compounds were responsible for the antimicrobial activity. Furthermore, in a 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging activity assay, only the visceral tissue extract showed activity. However, no extract had hemolytic activity against human red blood cells. Consequently, this study suggests the water-soluble extract of mottled anemone to be a promising source of proteinaceous antimicrobial compounds that can be utilized for development of novel antibiotics.

Radioprotective effects of delphinidin on normal human lung cells against proton beam exposure

  • Kim, Hyun Mi;Kim, Suk Hee;Kang, Bo Sun
    • Nutrition Research and Practice
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    • v.12 no.1
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    • pp.41-46
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    • 2018
  • BACKGROUND/OBJECTIVES: Exposure of the normal lung tissue around the cancerous tumor during radiotherapy causes serious side effects such as pneumonitis and pulmonary fibrosis. Radioprotectors used during cancer radiotherapy could protect the patient from side effects induced by radiation injury of the normal tissue. Delphinidin has strong antioxidant properties, and it works as the driving force of a radioprotective effect by scavenging radiation-induced reactive oxygen species (ROS). However, no studies have been conducted on the radioprotective effect of delphinidin against high linear energy transfer radiation. Therefore, this study was undertaken to evaluate the radioprotective effects of delphinidin on human lung cells against a proton beam. MATERIALS/METHODS: Normal human lung cells (HEL 299 cells) were used for in vitro experiments. The 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay assessed the cytotoxicity of delphinidin and cell viability. The expression of radiation induced cellular ROS was measured by the 2'-7'-dicholordihydrofluorescein diacetate assay. Superoxide dismutase activity assay and catalase activity assay were used for evaluating the activity of corresponding enzymes. In addition, radioprotective effects on DNA damage-induced cellular apoptosis were evaluated by Western blot assay. RESULTS: Experimental analysis, including cell survival assay, MTT assay, and Western blot assay, revealed the radioprotective effects of delphinidin. These include restoring the activities of antioxidant enzymes of damaged cells, increase in the levels of pro-survival protein, and decrease of pro-apoptosis proteins. The results from different experiments were compatible with each to provide a substantial conclusion. CONCLUSION: Low concentration ($2.5{\mu}M/mL$) of delphinidin administration prior to radiation exposure was radioprotective against a low dose of proton beam exposure. Hence, delphinidin is a promising shielding agent against radiation, protecting the normal tissues around a cancerous tumor, which are unintentionally exposed to low doses of radiation during proton therapy.

Increased Anticancer Activity by the Surfated Funcoidan from Korean Brown Seaweeds (한국산 길조류에서 추출한 Fucoidan의 황산기에 따른 항암작용)

  • Park, Jang-Su;Kim, An Deu Re;Kim, Eun-Hui;Seo, Hong-Suk;Choe, Won-Cheol
    • Journal of the Korean Chemical Society
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    • v.46 no.2
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    • pp.151-156
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    • 2002
  • Fucoidan is a kind of polysaccharides in brown seaweeds. For the past years have been extensively studied due to their numerous biological activities : anticancer, anticoagulant, antithrombotic, anti-inflammatory and antiviral. In this study, we h ave extracted fucoidan from the Korean brown seaweeds and examined it's anticancer activities for employed SV40 DNA replication assay, RPA-ssDNA binding assay of replication protein A(RPA: known as human single-stranded DNA-binding protein essential for DNA rep-lication) and MCF7 cell growth inhibition assay. In addition to, we found that chemically sulfated fucoidan'santicancer activity is more higher than natural and desulfated fucoidan. It seem that fucoidan's sulfate group affect on DNA replication, cause of decrease RPA's DNA binding activity. These results suggests that sulfated fucoidan from Korean brown seaweeds have anticancer activity.

Effects of Dendrobii herba and Punica granatum Extract on the Anti-oxidant, Anti-inflammatory, Anti-wrinkle and Whitening (석곡(石斛), 석류(石榴)의 항산화, 항염증, 주름, 미백에 미치는 영향)

  • HwangBo, Min;Roh, Seok-Sun;Seo, Hyeong-Sik
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.23 no.3
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    • pp.11-32
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    • 2010
  • Objective : The aim of this study is to determine the effects of Dendrobii herba extract and Punica granatum extract on skin disease and skin beauty. Methods : To investigate in vitro anti-oxidant activity assay, ethanol extracts of medicinal plants tested by DPPH radical, xanthine oxidase activity. In the next experiment, to investigate anti-inflammatory activity assay, examined by relations in NO synthesis, IL-$1{\beta}$, IL-6, TNF-${\alpha}$, NF-${\kappa}B$, COX-2, MAP kinase. To study Skin wrinkle formation effect, we were examined by tyrosinase activities, melanin synthesis in MNT-1 cell. Results : 1. In an anti-oxidant test, Dendrobii and Punica granatum extract showed high radical scavenging activity. 2. In an anti-inflammatory test, Dendrobii herba and Punica granatum extract weakly inhibited the lipopolysaccharide(LPS)-induced nitric oxide(NO) release from RAW 246.7 macrophage cells. Dendrobii herba and Punica granatum extract also inhibited LPS-induced IL-$1{\beta}$ and COX-2 expressions. The inhibitory effect of Dendrobii herba and Punica granatum extract on macrophage activation were via the inhibition of NF-${\kappa}B$, evidenced by transient transfection assay. however, Dendrobii herba and Punica granatum extract did not have any effects about activation of Jun-N-terminal kinase(JNK) and inhibition of p38 MAP kinase in RAW 264.7 cells. 3. In the skin wrinkle formation assay, Dendrobii herba and Punica granatum extract weakly inhibited collagenase and elastase, however it was not statistically significant. 4. In the skin whitening assay, Dendrobii herba and Punica granatum extract weakly inhibited tyrosinase activity, however, it was not statistically significant. They did not have any effect on melanin synthesis, indicating that they could not be applicable for skin whitening. Conclusion : Dendrobii herba extract and Punica granatum extract may play a significant role in skin disease and skin beauty.

Screening and Isolation of the Antitumor Agents from Medicinal Plants (I) (생약으로부터 항암성분의 검색 및 분리 (I))

  • Park, Shin-Young;Kim, Jin-Woong
    • Korean Journal of Pharmacognosy
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    • v.23 no.4
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    • pp.264-267
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    • 1992
  • The cytotoxic activity of medicinal plants was screened using A549 human lung cancer cell line. Plant materials were extracted with 80% methanol and fractionated to chloroform and water layers. Each methanol, chloroform, and water extract of thirty-two medicinal plants was tested for cytotoxic activity in A549 cell culture system and the cell viability was measured by SRB assay.

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