• Title/Summary/Keyword: Activity assay

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Detection of AluI Endonuclease Activity by Using Double Stranded DNA-Templated Copper Nanoclusters

  • Yang, Ji Su;Gang, Jongback
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.316-319
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    • 2021
  • Restriction endonucleases play an important role in molecular cloning, clinical diagnosis, and pharmacological drug studies. In this study, DNA-templated copper nanoclusters (DNA-CuNCs) were used to detect AluI endonuclease activity due to their high fluorescence emission and rapid synthesis of DNA-CuNCs under ambient conditions. Results showed that AluI activity was detected in a highly sensitive manner at low concentrations of AluI endonuclease by the fluorescence intensity of DNA-CuNCs. Additionally, its inhibition was monitored in the presence of daidzein under optimal conditions.

Evaluation of Anticancer Activity of Curcumin Analogues Bearing a Heterocyclic Nucleus

  • Ahsan, Mohamed Jawed
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.1739-1744
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    • 2016
  • We report herein an in vitro anticancer evaluation of a series of seven curcumin analogues (3a-g). The National Cancer Institute (NCI US) Protocol was followed and all the compounds were evaluated for their anticancer activity on nine different panels (leukemia, non small cell lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal cancer, prostate cancer and breast cancer) represented by 60 NCI human cancer cell lines. All the compounds showed significant anticancer activity in one dose assay (drug concentration $10{\mu}M$) and hence were evaluated further in five dose assays (0.01, 0.1, 1, 10 and $100{\mu}M$) and three dose related parameters $GI_{50}$, TGI and $LC_{50}$ were calculated for each (3a-g) in micro molar drug concentrations (${\mu}M$). The compound 3d (NSC 757927) showed maximum mean percent growth inhibition (PGI) of 112.2%, while compound 3g (NSC 763374) showed less mean PGI of 40.1% in the one dose assay. The maximum anticancer activity was observed with the SR (leukemia) cell line with a $GI_{50}$ of $0.03{\mu}M$. The calculated average sensitivity of all cell lines of a particular subpanel toward the test agent showed that all the curcumin analogues showed maximum activity on leukemia cell lines with $GI_{50}$ values between 0.23 and $2.67{\mu}M$.

A Study on the Remaining Concentration of Pesticides in Tap Water of Taejon City by Ellman′s Enzyme Method and the Countermeasure (Ellman 효소법에 의한 대전시 상수도내 살충제의 잔류농도 결정 및 그 대책에 관한 연구)

  • 이봉호;이영순;전종한
    • Journal of Environmental Science International
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    • v.8 no.1
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    • pp.19-26
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    • 1999
  • The degree of pesticides accumulation in tap water in Taejon from June 1995 to Apr 1996 was measured by Ellman's coupled enzyme assay. Since organic phosphate and carbamate pesticides specifically inhibit the neurotransmitter modulating enzyme acetylcholinesterase(AChE), the enzyme activity can be used as a diagnosis for the pesticides accumulation in water and various samples. During the period of this study, the enzyme activity was changed almost every week. The lowest enzyme activity was 64 % of that of the control reaction and there are several days showing about 100 % enzyme activity. In general, the enzyme activity is higher in summer than other seasons especially early spring times. The pH value of tap water was very close to neutral(pH 7.0) and it seems that the enzyme activity was not affected by the small pH changes. Either boiling of tap water or addition of NaOH solution decomposed the pesticide components. These results show that AChE assay is a convenient, sensitive, and reliable method for detection of pesticides in water samples.

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Screening and Evaluation of Antibacterial Metabolites from Entomopathogenic Fungi

  • Shin, Tae-Young;Ko, Seung-Hyun;Lee, Won-Woo;Bae, Sung-Min;Choi, Jae-Bang;Woo, Soo-Dong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.26 no.2
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    • pp.89-94
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    • 2013
  • Entomopathogenic fungal species have been investigated for their potential use as biological control agents owing to their natural role as insect pathogens. These fungi produce a wide range of secondary metabolites with high therapeutic values, such as antibiotics and cytotoxic substances. To evaluate the antibacterial activity of entomopathogenic fungi, 10 isolates from Korean soil were selected and tested for their activity against Escherichia coli by using fungal culture filtrates. Antibacterial activity was assessed using a two-step process: (1) a screening assay for the selection of fungal isolates and (2) a quantitative assay to evaluate the activity of select fungi. Although 4 fungal isolates were selected through the screening assay, only 3 fungal isolates, from Beauveria bassiana and Metarhizium anisopliae, showed high antibacterial activity according to the quantitative assay. The antibacterial activity of selected fungal culture filtrates was stable when exposed to heat and proteolytic enzyme treatments, which indicated that the antibacterial compound is not a protein. These entomopathogenic fungal metabolites might be useful as a source for bacterial control and in the pharmaceutical industry.

Inhibitory Effects of Actinidia arguta on HIV-1 Reverse transcriptase, HIV-1 Protease and alpha-glucosidase in vitro and in silico (다래나무 추출물의 HIV-1 효소억제활성과 구조활성상관(QSAR)예측)

  • Yu, Young-Beob
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.115-121
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    • 2006
  • Objectives : For the purpose of developing new anti-HIV agents from natural sources, the extracts of Actinidia arguta were tested for their inhibitory effects on essential enzymes as the reverse transcriptase (RT), protease and ${\alpha}-\;glucosidase$. And we predicted inhibition activity of major compounds of Actinidia arguta using Quantitative Structure Activity Relationships (QSAR). Methods : In this assay the activity of HIV-1 reverse transcriptase is measured as the formation of a strand of copy-DNA (cDNA) using RNA as a template. The activity of HIV-1 protease is measured as the cleavage of an oligopeptide by HIV-1 protease. Results : In the anti-HIV-1 RT using Enzyme Linked Oligonucleotide Sorbent Assay (ELOSA) method, water extracts (100ug/ml) of stem and leaf showed strong activity of 93.9% and 91.9%, respectively. In the HIV-1 protease inhibition assay, aqueous stem extract inhibited the activity of the enzyme to cleave an oligopeptide, resembling one of the cleavage sites in the viral polyprotein which can only be processed by HIV-1 protease with 56.8%. In the ${\alpha}-glucosidase$ inhibition assay, aqueous stem extract showed activity of 73.1%. Conclusion : We found out this result, for these samples it is possible that the inhibition of the viral replication in vitro is due to the inhibition at least one of RT and ${\alpha}-glucosidase$. It would be of great interest to identify the compounds which are responsible for this inhibition, since all therapeutically useful agent up to date are RT, PR and ${\alpha}-glucosidase$ inhibitors.

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Screening of Anti-HIV-1 Activity of Natural Products by MTT Assay (MTT Assay에 의한 천연물질의 항 HIV-1 활성 검색)

  • Lee, Joo-Shil;Nam, Jeong-Gu;Kang, Chon;Lee, Hong-Rae;Lee, Young-Jong;Shin, Yung-Oh
    • The Journal of Korean Society of Virology
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    • v.27 no.1
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    • pp.87-95
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    • 1997
  • Methanol and/or boiling water extraction of 201 natural products and subsequent MTT assay using MT-4 cell line was carried out to screen the anti-HIV-1 activity. Among 97 methanol extracts, 7 extracts from Chrysanthemi Indicium Flos, Magnoliae Cortex, Machili Cortex, Reynoutriae Rhizoma, Lithospermi Radix, Agastachis Herba, and Chaenomelis Fructus showed anti-HIV-1 activity and their SI value were 2.25 to 5.77. In addition, among 119 boiling water extracts, 10 extracts from Lonicerae Caulis et Foloium, Elsholtziae Herba, Leonuri Herba, Portulacae Herba, Schizonepetae Herba, Curcumae Rhizoma, Amomi Cardamomi Fructus, Cirsii Radix et Herba, Carpesii Herba, and Siegesbeckiae Herba showed anti-HIV-1 activity and their SI value were 1.30 to 7.64. Methanol extracts of above seven natural products were fractionated and the anti-HIV-1 activity of each fraction was examined. Extraction was carried out with hexane, chloroform, butanol, and water to trace active anti-HIV-1 componets. As a result, the water fraction of Magnoliae Cortex, Machili Cortex, Reynoutriae Rhizoma, Agastachis Herba, Chaenomelis Fructus and the butanol fraction of Chrysanthemi Indicium Flos, Reynoutriae Rhizoma showed anti-HIV-1 activity and their SI value were 1.40 to 8.02. We could reach a conclusion that studies to trace the anti-HIV-1 active component of each natural products in further fractionation and to identify its structure by Infrared spectroscopy, NMR spectroscopy and gel permeation chromatography were needed.

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Cosmetic Effects of Dietary Fiber from Mozuku, Cladosiphon novae-caledoniae Kylin (큰실말(Cladosiphon novae-caledoniae Kylin) 부산물의 화장품소재 특성)

  • Kim, In Hae;Lee, Jae Hwa
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.3
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    • pp.285-295
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    • 2016
  • To investigate the effect of dietary fiber from mozuku, Cladosiphon novae-caledoniae kylin (C. novae-caledoniae kylin) on the skin care, we measured anti-oxidant activity, anti-microbial activities, tyrosinase activity inhibition and elastic activity. B16F10 melanoma cell (MTT assay) were used to measure cell viability. MC and MI exhibited in vitro antibacterial activity against Staphyloccus aureus (S. aureus) and MRSA without antifungal activity. Mozuku extract (MS) showed excellent tyrosinase inhibition effect compared to arbutin as a positive control (to 49% for tyrosine). The wrinkle-improving effect was relatively low. However, wrinkle-improving effect was relatively low. DPPH free radical scavenging activity was 89% in a concentrations at $500{\mu}g/mL$. These results indicate that the mozuku extracts may be an effective cosmetic ingredient for skin whitening.

Activity of Alkaline Phosphatase from the Mosquito, Culex pipiens pallens (홍모기(Culex pipiens pallens)의 난성숙 과정 중 alkaline phosphase의 활성)

  • 이영수;이승훈;박영민;성기창
    • The Korean Journal of Zoology
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    • v.36 no.3
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    • pp.425-432
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    • 1993
  • Alkaline phosphatase from Culex pipiens pallens was examined to determine the optimal assay condition and to assay the activity during ovarian development. The activity of alkaline phosphatase in a male and a nongravid female continuously were declined after eclosion. But by the stimulus of a blood meal, the enzyme activity was increased dramatically. At 30 hr. after a blood meal, the maximal activity was reached and then declined. And after 48 hr. after a blood meal, the second activity increase was revealed. This second increase was maintained up to oviposition. The first activity increase was revealed in the midgut and the second increase was done in the ovary to assay the organ distribution of alkaline phosphatase. In electrophresis data, it was shown 5 isozyme bands, ALP-1 and ALP-2 in the ovary, ALP-3 in the thorax and the midgut, and ALP-4 and ALP-5 in the thorax, the fatbody and the midgut in crude extract at 30 hr. after a blood meal. One the same ovary pattern were shown at 72 hr. after a blood meal.

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In vitro Biological Activity Assay of Ethanol Extract of Radish (무 에탄올 추출물의 in vitro 생리활성 분석)

  • Jung, Min-Suk;Lee, Gun-Soon;Chae, Hee-Jeong
    • Applied Biological Chemistry
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    • v.47 no.1
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    • pp.67-71
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    • 2004
  • In vitro biological activities of ethanol extract of radish including whitening, hangover removal, antimicrobial and antioxidant activities were analyzed. For whitening activity assay, tyrosinase inhibition rate was measured as $IC_{50}$ (50% inhibitory concentration). The $IC_{50}$ values of radish trunk and root extracts were estimated as 0,9 mg/ml and 2.1 mg/ml, respectively. Radish trunk extract showed 2.5-fold tyrosinase inhibition activity of radish root extract, however, there was no significant difference according to radish species. By alcohol dehydrogenase (ADH) activity assay as a hangover removal activity assay, radish trunk extract (2.5 mg/ml) and root extracts (8 mg/ml) showed ]50% activation of ADH. TBA values of radish trunk and root extracts (1% of each) were 43-61 % level of ${\alpha}-tochoperol$ (2.2%). From the analysis of in vitro biological activities of radish, it was suggested that radish could be used in functional food or cosmetics containing hangover removal, whitening and antioxidant activities.

Anti-invasive Effect of Artemisia scoparia Halophyte Extract and its Solvent-partitioned Fractions in Human Fibrosarcoma Cells (인간 섬유육종세포에서 비쑥 추출물과 유기용매 분획물의 암전이 억제 효과)

  • Kim, Junse;Kong, Chang-Suk;Sim, Hyun-Bo;Seo, Youngwan
    • Journal of Life Science
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    • v.31 no.12
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    • pp.1100-1109
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    • 2021
  • The halophyte Artemisia scoparia is an edible medicinal plant, with insecticidal, anti-inflammatory, anticholesterol, antipyretic, and antibacterial effects. The aim of this study was to assess the inhibitory effect of crude extract and solvent-partitioned fractions obtained from A. scoparia on MMP-2 and MMP-9 activity in phorbol-12-myristate-13-acetate (PMA)-stimulated human fibrosarcoma HT-1080 cells using four different activity tests: gelatin zymography, MMP enzyme-linked immunosorbent assay (ELISA), wound healing assay, reverse transcription-polymerase chain reaction (RT-PCR), and Western blot assay. A. scoparia samples were extracted twice with methylene chloride (MC) and twice with methanol (MeOH). After the MC and MeOH crude extracts were combined, the combined crude extracts showed a significant inhibitory effect against MMP-2 and MMP-9 enzymes. They were then fractionated into n-hexane, 85% (v/v) aqueous methanol (85% (v/v) aq.MeOH), n-butanol, and water according to solvent polarity. Among the four solvent-partitioned fractions, n-hexane and 85% (v/v) aq. MeOH fractions significantly inhibited MMP-2 and MMP-9 activity and cell mobility. In addition, the n-hexane and 85% (v/v) aq.MeOH fractions effectively inhibited MMP-2 and -9 activity in the gelatin zymography and MMP ELISA assay. In the wound healing assay, RT-PCR, and Western blot assay, all solvent-partitioned fractions, except the H2O fraction, significantly suppressed cell migration, as well as the expression levels of MMP-2 and -9 mRNA and proteins.