• Title/Summary/Keyword: Activity assay

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Antioxidant and Quinone Reductase Inductive Activities of Various Organs of Pepper (고추 기관별 항산화 활성과 quinone reductase 유도활성)

  • Ku, Kang-Mo;Kang, Young-Hwa
    • Journal of Applied Biological Chemistry
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    • v.53 no.1
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    • pp.31-36
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    • 2010
  • We analyzed antioxidant and quinone reductase (QR) inductive activities of various organs of pepper for utilizing by-product of them. Peppers were separated into fruits, roots, stems, and leaves and extracted with methanol for the analysis. As a result, pepper leaves showed higher phenol content than other organs. Using the DPPH assay, there was not considerably different activity depending on pepper organs, but pepper leaves showed significantly higher antioxidant activity using the ABTS assay. In FTC and TBA assay, stems and leaves showed significantly higher lipidperoxidation inhibitory activity. In QR inductive assay, pepper tissues showed different QR inductive activity: leaves>roots>>stems>fruits. In addition, pepper leaves showed highest antiproliferation activity on hepa1c1c7 among pepper tissues in $50-200\;{\mu}g/mL$. These results indicate that pepper leaves have high potential to be a good functional food material due to high QR inductive and antioxidant activities.

Apoptosis-Inducing Activity of HPLC Fraction from Voacanga globosa (Blanco) Merr. on the Human Colon Carcinoma Cell Line, HCT116

  • Acebedo, Alvin Resultay;Amor, Evangeline Cancio;Jacinto, Sonia Donaldo
    • 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.

In Vitro Studies on Phytochemical Content, Antioxidant, Anticancer, Immunomodulatory, and Antigenotoxic Activities of Lemon, Grapefruit, and Mandarin Citrus Peels

  • Diab, Kawthar AE
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3559-3567
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    • 2016
  • Background: In recent years, there has been considerable research on recycling of agro-industrial waste for production of bioactive compounds. The food processing industry produces large amounts of citrus peels that may be an inexpensive source of useful agents. Objective: The present work aimed to explore the phytochemical content, antioxidant, anticancer, antiproliferation, and antigenotxic activities of lemon, grapefruit, and mandarin peels. Materials and Methods: Peels were extracted using 98% ethanol and the three crude extracts were assessed for their total polyphenol content (TPC), total flavonoid content (TFC), and antioxidant activity using DPPH (1, 1-diphenyl-2-picrylhydrazyl). Their cytotoxic and mitogenic proliferation activities were also studied in human leukemia HL-60 cells and mouse splenocytes by CCK-8 assay. In addition, genotoxic/antigenotoxic activity was explored in mouse splenocytes using chromosomal aberrations (CAs) assay. Results: Lemon peels had the highest of TPC followed by grapefruit and mandarin. In contrast, mandarin peels contained the highest of TFC followed by lemon and grapefruit peels. Among the extracts, lemon peel possessed the strongest antioxidant activity as indicated by the highest DPPH radical scavenging, the lowest effective concentration 50% ($EC_{50}=42.97{\mu}g\;extract/mL$), and the highest Trolox equivalent antioxidant capacity (TEAC=0.157). Mandarin peel exhibited moderate cytotoxic activity ($IC_{50}=77.8{\mu}g/mL$) against HL-60 cells, whereas grapefruit and lemon peels were ineffective anti-leukemia. Further, citrus peels possessed immunostimulation activity via augmentation of proliferation of mouse splenocytes (T-lymphocytes). Citrus extracts exerted non-cytotoxic, and antigenotoxic activities through remarkable reduction of CAs induced by cisplatin in mouse splenocytes for 24 h. Conclusions: The phytochemical constituents of the citrus peels may exert biological activities including anticancer, immunostimulation and antigenotoxic potential.

Biological Activity and Inhibition of Non-Enzymatic Glycation by Methanolic Extract of Rosa davurica Pall. Roots

  • Hu, Weicheng;Han, Woong;Jiang, Yunyao;Wang, Myeong-Hyeon;Lee, Young-Mee
    • Preventive Nutrition and Food Science
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    • v.16 no.3
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    • pp.242-247
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    • 2011
  • The methanolic extract of Rosa davurica Pall. roots exhibited strong antioxidant activity in a 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging assay and was found to be a dose-dependent inhibitor of non-enzymatic formation of advanced glycation end products (AGEs), which are relevant to diabetes complications. HPLC-diode array detector (DAD) analysis of the R. davurica Pall. root extract led to the identification of four compounds: hydrocaffeic acid, catechin, epicatechin, and ellagic acid. Catechin was present in the largest amount and exhibited high antiglycation activity. A CYP3A4 assay was used to investigate potential interactions between drugs and the extract, and results suggest that the R. davurica Pall. root extract had moderate potential for interfering with drug metabolism. The R. davurica Pall. extract did not display anti-inflammatory activity on the level of that for tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in a lipopolysaccharide (LPS)-stimulated macrophage assay; however, the extract did exhibit low to moderate immunostimulatory activity in a pro-inflammatory macrophage assay. Therefore, we conclude that R. davurica Pall. root is a promising anti-AGE agent with low to moderate risks of associated inflammation or drug interaction.

Effect of Five Korean Native Taraxacum on Antioxidant Activity and Nitric Oxide Production Inhibitory Activity (국내 자생 민들레 5종의 항산화 활성 및 Nitric Oxide 생성억제 활성)

  • Choi, Kyeong Hee;Nam, Hyeon Hwa;Choo, Byung Kil
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.3
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    • pp.191-196
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    • 2013
  • The effect on the antioxidant activity and Nitric Oxide activity production inhibitory activity of Taraxacum has not been known. Therefore, phenolics and flavonoid contents were investigated from the ethanol extracts of five different Taraxacum species. The results showed that, among the five Taraxacum, T. hallaisanensis contains the highest total phenolic and flavonoid contents. When the antioxidant activity was measured by DPPH, $ABTS^+$ and reducing power activity, the free radical scavenging activity of T. hallaisanensis was also the highest among five Taraxacum species. However, measurement by CCK-8 assay in Raw264.7 cells indicated that the extracts of Taraxacum species have no effect on cell viability. Moreover, we also investigated the effect of Taraxacum species on NO scavenging activity in lipopolysaccharide (LPS)-stimulated Raw264.7 cells. The results clearly showed that Taraxacum species inhibited NO production, and the inhibitory effect of T. hallaisanensis was the strongest. The above results suggested that Taraxacum species affected the antioxidant and NO scavenging activity, and among the five species, antioxidant and NO scavenging activity assay of T. hallaisanensis was significantly higher than those of other four Taraxacum species. Therefore, T. hallaisanensis could be used as a potential drug with anti-oxidant and anti-inflammatory effect.

Antioxidative and Antibacterial Activity and Tyrosinase Inhibitory Activity of the Extract and Fractions from Taraxacum coreanum Nakai (흰민들레 에탄올 추출물 및 분획물의 항산화 및 항균활성과 Tyrosinase 저해활성)

  • Im, Do-Youn;Lee, Kyoung-In
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.4
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    • pp.238-245
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    • 2011
  • In this study, we investigated antioxidative activity, antibacterial activity against pathogenic strains including methicillin resistant Staphylococcus aureus (MRSA), and tyrosinase inhibitory activity in 75% ethanol extract of Taraxacum coreanum and its fractions. The total polyphenol and flavonoid contents of the extract were 238.59mg/g and 33.18mg/g and the total polyphenol and flavonoid contents of the ethyl acetate fraction were 427.81mg/g and 148.90mg/g as the highest content of fractions. In DPPH radical scavenging ability, $SC_{50}$ values of the ethyl acetate and butanol fraction were 38.40${\mu}g/ml$ and 82.28 ${\mu}g/ml$, respectively. In antibacterial activity by the disc diffusion assay against S. aureus, S. epidermidis and MRSA, the ethyl acetate fraction showed stronger antibacterial activity than other fractions and the extract. Especially, the ethyl acetate fraction was exhibited effective antibacterial activity against MRSA. In the cytotoxicity measurement by MTT assay, the extract and fractions were exhibited Raw 264.7 cell viabilities of 96.32~143.21% as nontoxic result in concentration of 5~100 ${\mu}g/ml$. As a result, the ethyl acetate fraction of the 75% ethanol extract from T. coreanum could be applicable to functional materials for related fields.

Measurement of Antioxidant Activity of Anserine, Taurine, and L-Histidine in vitro and Content of Anserine, Taurine, and L-Histidine in Mature and Juvenile Rainbow Trout (Onchorhynchus mykiss) Muscle

  • Yun-Hee chio;Kim, Harriet
    • Preventive Nutrition and Food Science
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    • v.1 no.2
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    • pp.174-178
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    • 1996
  • The content of anserine, taurine, and L-histidine was measured by HPNC in the muscle of mature(670~690g) and juvenile(80~120g) rainbow trout fatmed in Chungsun, Korea. The concentration of anserine and taurine was higher in mature rainbow trout than in juvenile, but that of L-histidine was lower in mature than in juvenile. When measured with the chemiluminescence(CL) assay, anserine and taurine showed very powerful antioxidative activity above physiological concentration rainbow trout. Taurine still showed antioxidative activity below physiological concentration, while anserine showed prooxidative activity below that. L-Histidine was prooxidative dose-dependently. In TBA method, while taurine showed very week antioxidative effect, anserine appeared very powerful antioxidant and L-histidine prooxidant at physiological concentration. There was no synergism between anserine and taurine and anserine inhibited prooxidative effect of L-histidine.

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Cytotoxicity and L-Amino Acid Oxidase Activity of Crude Insect Drugs

  • Ahn, Mi-Young;Ryu, Kang-Sun;Lee, Yong-Woo;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • v.23 no.5
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    • pp.477-481
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    • 2000
  • The cytotoxicity of crude insect drugs was measured using HeLa cells originating from human cervix and uterine cancer. using the dye uptake assay in order to find potential anticancer agents. Three kinds of extracts (buffer, methanol and ethylacetate) were prepared from 26 insects and used as raw materials for the activity assay. Among these, the buffer extracts from Tabanus, Mylabris and Huechys showed a potent anticancer activity, and those from Catharsius, Red ant, Scorpion, Tabanus and Vespae Nidus showed a strong L-amino acid oxidase (AAO) activity as well as cytotoxicity. In contrast, buffer extracts from Gryllotalpa orientalis and Apriona germari larvae showed greater/more rapid Hela cell growth than that of other insects.

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Isolation and Biological Activity of Verbascoside, A Potent Inhibitor of Protein Kinase C from the Calyx of Campsis grandiflora (능소화의 꽃받침으로부터 Protein Kinase C 저해물질인 Verbascoside의 분리 및 그 생물활성)

  • 이현선;박문수;오원근;안순철;김보연;김환묵;오구택;민태익;안종석
    • YAKHAK HOEJI
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    • v.37 no.6
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    • pp.598-604
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    • 1993
  • The calyx extract of Campsis grandiflora displayed inhibitory activity against protein kinase C from the bovine brain. Separation guided by protein kinase C enzyme assay and bleb forming assay led to isolation of a potent protein kinase C inhibitor that was identified as a known phenylpropanoid glycoside, verbascoside. It suppressed completely bleb-formation of K562 cell surface induced by phorbol 12,13-dibutylate at the concentration of 60 $\mu\textrm{g}$/ml and IC$_{50}$ of the protein kinase C occured at 20 $\mu{M}$. This compound was tested for cytotoxic activity against ten human tumor cell lines in vitro. it exhibited moderate cytotoxic activity against skin tumor cell line M14 (IC$_{50}$ 2.2 $\mu\textrm{g}$/ml) and very weak cytotoxicity against other cell lines (IC$_{50}$>10 $\mu\textrm{g}$/ml)

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The Inhibitory Effects of Cannabinoids of Cannabis Against Human Skin Melanoma Cells (마자인의 카나비노이드가 인체피부흑색종세포에 미치는 억제효과)

  • Lee Ki-Nam;Kim Meung-Shin;Back Seung-Hwa
    • Journal of Society of Preventive Korean Medicine
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    • v.2 no.1
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    • pp.13-30
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    • 1998
  • Geraniol (1), olivetol (2), cannabinoids (3 and 4) and 5-fluorou.a.il (5). were tested for their growth inhibitory effects against SK-MEL-3 cell lines using two different 3-(4,5-dimethylthiazol- 2yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. These compounds showed inhibitory activity in vitro in the micromolar range against SK-MEL-3 cell lines. In general, the antitumor activity of these compounds (1, 2, 3, 4 and 5) was in a dose-dependent over the micromolar concentration range $1\;to\;100{\mu}M$. The comparison of $IC_{50}$ values of these compounds in tumor cell lines shows that their susceptibility to these compounds decrease in the following order : OLVTL > CBG > CBD > 5-FU > CRNL in MTT assay, CBG > OLVTL > CBD > GRNL > 5-FU in SRB assay. Cannabinoids (3 and 4) and 5-fluorouracil (5) were tested for their cytotoxic effects on NIH 373 fibroblasts using two different MTT assay and SRB assay. These compounds exhibited potent cytotoxic activities in vitro in the micromolar range against NIH 373 fibroblasts. In general, the cytotoxic activities of these compounds (3, 4 and 5) were in a dose-dependent over the micromolar concentration range $1\;to\;100{\mu}M$. The comparison of $CD_{50}$ values of these compounds on NIH 373 fibroblasts shows that their susceptibility to these compounds decrease on the following order ; CBD > 5-FU > CBG in MTT assay and SRB assay. Cannabigerol (3) was shown the least cytotoxic activity on NIH 373 fibroblasts. Cannabigerol (3) exhibited the most growth-inhibitory activity against SK-MEL-3 cell lines.

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