• Title/Summary/Keyword: Chemical assay

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Antioxidant Activity, Total Phenolics, Vitamin C and Sugar Content during Fruit Ripening of Five Different Jujube Cultivars (대추(Zyziphus jujuba) 품종별 성숙정도에 따른 항산화 활성, 총페놀 함량, 비타민 C 함량 및 당함량)

  • Park, Youngki;Kim, Jae-Hee
    • Korean Journal of Plant Resources
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    • v.29 no.5
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    • pp.539-546
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    • 2016
  • The aim of this study was to analyze antioxidant activity, total phenolics, and vitamin C content in unripe and ripe jujube cultivar and to investigate correlation between antioxidant and chemical constituents including total phenolics and vitamin C. The antioxidant activity was measured by the DPPH radical scavenging activity, ABTS radical scavenging activity, and reducing power. Although, the weight, diameter and soluble solids of Zyziphus jujuba fruit were increased with the progress of ripening, antioxidant activity and total phenolics were decrease during ripening. The highest free-radical scavenging activity (at 2.5 ㎎/㎖) and reducing power (at 1 ㎎/㎖) in fruit were 81.75% (Bokjo, unripe) and 0.80 (Mudeung, unripe), respectively. The highest total phenolic content in unripe fruit of Mudeung was 62.50 ㎍/g. A linear correlation (r=0.911) was shown between DPPH radical scavenging activity and total phenolic content.

Rare ginsenoside Ia synthesized from F1 by cloning and overexpression of the UDP-glycosyltransferase gene from Bacillus subtilis: synthesis, characterization, and in vitro melanogenesis inhibition activity in BL6B16 cells

  • Wang, Dan-Dan;Jin, Yan;Wang, Chao;Kim, Yeon-Ju;Perez, Zuly Elizabeth Jimenez;Baek, Nam In;Mathiyalagan, Ramya;Markus, Josua;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.42 no.1
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    • pp.42-49
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    • 2018
  • Background: Ginsenoside F1 has been described to possess skin-whitening effects on humans. We aimed to synthesize a new ginsenoside derivative from F1 and investigate its cytotoxicity and melanogenesis inhibitory activity in B16BL6 cells using recombinant glycosyltransferase enzyme. Glycosylation has the advantage of synthesizing rare chemical compounds from common compounds with great ease. Methods: UDP-glycosyltransferase (BSGT1) gene from Bacillus subtilis was selected for cloning. The recombinant glycosyltransferase enzyme was purified, characterized, and utilized to enzymatically transform F1 into its derivative. The new product was characterized by NMR techniques and evaluated by MTT, melanin count, and tyrosinase inhibition assay. Results: The new derivative was identified as (20S)-$3{\beta},6{\alpha},12{\beta}$,20-tetrahydroxydammar-24-ene-20-O-${\beta}$-D-glucopyranosyl-3-O-${\beta}$-D-glucopyranoside(ginsenoside Ia), which possesses an additional glucose linked into the C-3 position of substrate F1. Ia had been previously reported; however, no in vitro biological activity was further examined. This study focused on the mass production of arduous ginsenoside Ia from accessible F1 and its inhibitory effect of melanogenesis in B16BL6 cells. Ia showed greater inhibition of melanin and tyrosinase at $100{\mu}mol/L$ than F1 and arbutin. These results suggested that Ia decreased cellular melanin synthesis in B16BL6 cells through downregulation of tyrosinase activity. Conclusion: To our knowledge, this is the first study to report on the mass production of rare ginsenoside Ia from F1 using recombinant UDP-glycosyltransferase isolated from B. subtillis and its superior melanogenesis inhibitory activity in B16BL6 cells as compared to its precursor. In brief, ginsenoside Ia can be applied for further study in cosmetics.

Isolation and Identification of Antioxidants from Methanol Extract of Sword Bean (Canavalia gladiata) (작두콩의 메탄올 추출물로부터 항산화 활성 화합물의 분리 및 동정)

  • Kim, Jong-Pil;Lee, Hyang-Hee;Moon, Jae-Hak;Ha, Dong-Ryong;Kim, Eun-Sun;Kim, Jin-Hwan;Seo, Kye-Won
    • Korean Journal of Food Science and Technology
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    • v.45 no.6
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    • pp.777-784
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    • 2013
  • The ethyl acetate (EtOAc) layer of Canavalia gladiata (sword bean) methanol extracts showed higher 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity than other layers. Four phenolic compounds were isolated from the EtOAc layer by silica gel column chromatography and prep-HPLC using a guided DPPH radical-scavenging assay. The isolated compounds were identified as methyl gallate (1), gallic acid (2), 1,6-di-O-galloyl ${\beta}$-$\small{D}$-glucopyranoside (3), and 1,4,6-tri-O-galloyl ${\beta}$-$\small{D}$-glucopyranoside (4) based on MS and NMR analyses. Among the four compounds, no. 4 was isolated from this plant for the first time. Their DPPH radical-scavenging activities based on $SC_{50}$ decreased in the following order: 4 (6.9 ${\mu}M$)>3 (8.3 ${\mu}M$)>2 (10.0 ${\mu}M$)>1 (10.3 ${\mu}M$).

Accumulation of Microcystins in Fish and Evaluation of Potential Human Health Risks: A Case Study on a Eutrophic Reservoir in Korea (마이크로시스틴의 어류내 축적성 및 인체 위해성 평가: 국내 저수지 사례연구)

  • Yoon, Hyojung;Seo, Jungkwan;Kim, Taksoo;Jo, Areum;Kim, Jungkon;Lee, Doohee;Kim, Pilje;Choi, Kyunghee
    • Journal of Environmental Health Sciences
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    • v.42 no.1
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    • pp.10-18
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    • 2016
  • Objectives: Microcystin (MC) produced during cyanobacterial blooms is a worldwide problem presenting a serious health threats to humans and ecosystems. During July through October of 2013, the Ilwol Reservoir experienced a high biomass of phytoplankton (maximum $211.7mg/m^3$ of Chlorophyll-a) containing the toxigenic cyanobacterium Oscillatoria sp. The aim of this study is to analyze MC concentration in the reservoir water, as well as in representative fish species (Carassius cuvieri, Carassius auratus, Channa argus). We also evaluated the human health risk of exposure to MCs accumulated in the fish. Methods: Concentrations of MCs in the water and fish samples were analyzed by liquid chromatography with a triple quadrupole tandem mass spectrometer (LC/MS/MS) and enzyme-linked immunosorbent assay (ELISA). Results: The total levels of four MC variants, including MC-LR, MC-RR, MC-YR and MC-LA were below the WHO drinking water guideline limit (1 ug MC-LR per liter) both for the dissolved and particulate fraction present in the water samples. The mean MC concentrations in the livers of all species were significantly higher than in the gills (p < 0.01) and muscles (p < 0.05). The values of estimated daily intake of MCs in muscles, the edible part of the fish, would be only $0.005-0.015{\mu}g/kg{\cdot}day$, much lower than WHO's provisional tolerable daily intake of $0.04{\mu}g/kg{\cdot}day$. Conclusion: This study suggests that, owing to the spatial distribution or temporal variation of MC, there is a need for careful monitoring of cyanotoxin in reservoir water and aquatic animals to protect public health.

CA 15-3 (Mucin-1) and Physiological Characteristics of Breast Cancer from Lahore, Pakistan

  • Begum, Mumtaz;Karim, Sajjad;Malik, Arif;Khurshid, Rukhshan;Asif, Muhammad;Salim, Asmat;Nagra, Saeed Ahmed;Zaheer, Ahmad;Iqbal, Zafar;Abuzenadah, Adel Mohammed;Alqahtani, Mohammed Hussain;Rasool, Mahmood
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.5257-5261
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    • 2012
  • Background: High incidence of breast cancer and its fatal effect has reached an alarming stage across the globe, including the third world countries. Many factors have been reported to be associated with the development of breast cancer but detailed structural and functional information is missing. CA 15-3 is one of the known potential tumor marker of breast cancer; however little is known about structure and functional site of this protein. Present study aims to investigate the functional role of CA 15-3 in breast cancer, especially in development and metastasis. Material and Methods: Hundred female breast cancer patients confirmed by histopathological reports were included in the study. Their physiological characters were recorded in a performa. Enzyme linked immunosorbent assay (ELISA) technique was used to estimate serum CA 15-3 level. Immunohistochemistry was done for estrogen (ER), progesterone (PR) and Her2/neu receptors expression. Results: The study revealed the details of physiological characteristics of female breast cancer. Mean age was $37.72{\pm}5.99$ and $55.05{\pm}7.28$ years and serum CA 15-3 (MUC1) level was $60.47{\pm}8.59$ and $63.17{\pm}4.58$ U/ml in pre and post-menopause respectively, and both groups of women had sedentary life style. Their receptor status especially of progesterone, estrogen and HER-2/neu were positive in 50% of premenopausal women and 65% of postmenopausal women. Conclusion: There are multiple physiological factors promoting breast cancer. High serum CA 15-3 level and hormonal imbalance of ER, PR and Her2/neu appears to be the main cause of breast cancer. It may be possible that the functional sites of these proteins may be altered which may increase the chances of metastasis in breast cancer.

Expression of CP4 5-Enol-Pyruvylshikimate-3- Phosphate Synthase Transgene in Inbred Line of Korean Domestic Maize (Zea may L.) (국내 옥수수 순계주에서 CP4 5-Enol- Pyruvylshikimate-3- Phosphate Synthase 유전자의 발현)

  • Cho, Mi-Ae;Kwon, Suk-Yoon;Kim, Jin-Seog;Lee, Byoung-Kyu;Moon, Choo-Yeun;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.375-380
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    • 2007
  • This study was conducted to develop herbicide-resistance domestic maize plants by introducing the CP4 5-enol-pyruvylshikimate-3-phosphate synthase (CP4 EPSPS) gene using Agrobacterium tumefaciens-mediated immature embryo transformation. Immature embryos of five genotypes (HW1, KL103, HW3, HW4, HW7) were co-cultivated with strains Agrobacterium tumefaciens (strain C58C1) containing the binary vector (pCAMBIA2300) carrying Ubiquitin promoter-CP4 EPSPS gene and Cauliflower mosaic virus 35S (CaMV35S) promoter-nptll gene conferring resistance to paromomycin as a selective agent. The presence and expression of CP4 EPSPS transgene were confirmed by PCR, RT-PCR and Northern blot analysis, respectively. Also, the resistance to glyphosate in the transgenic maize ($T_1$) was analyzed by shikimate accumulation assay. The frequency (%) of paromomycin-resistance callus was 0.37, 0.03, 2.20, 2.37, and 0.81% in pure lines HW1, KL103, HW3, HW4 and HW7, respectively. EPSP transgene sequences were amplified in putative transgenic plants that regenerated from paromomycin-resistance calli of two inbred lines (HW3, HW4). Of them, RT-PCR and Northern blot analyses revealed that the transgene was only expressed in two transgenic events (M266, M104) of HW4 inbred line, and a mild glyphosate resistance of transgenic event (M266) was confirmed by the lower shikimate accumulation in leaf segments. These results demonstrate that transgenic maize with herbicide-resistance traits in Korean genotype can be genetically obtained.

Studies on Analysis Method of T-2 Toxin by ELISA (ELISA에 의한 T-2 toxin의 분석법에 관한 연구)

  • 오유진;장성재;윤여표
    • Journal of Food Hygiene and Safety
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    • v.3 no.2
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    • pp.65-73
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    • 1988
  • T-2 toxin is one of mycotoxins produced by fungi such as Fusarium spp. and possesses a potent cytotoxicity to eukaryotic cell. The contamination of mycotoxins in cereals and feedstuffs is one of the great concerns in health authorities. Therefore, the development of the specific, sensitive and simplified analysis method for T -2 toxin is required. During more than ten years, several chemical and biological analysis methods were proposed and applied for the detection and quantification of T-2 toxin. TLC, GLC-FID and GC-MS are widely employed, but these methods required numerous clean-up procedures before analysis, and the detection limit for T-2 toxin is more than 10 ppb. Biological analysis methods with dermal tissues and cultured cells are not specific to T-2 toxin, since T-2 toxin and other related derivatives possess a similar toxicological activity although their relative activity is different each otber. Based on tbe specific reaction between antibody and antigen, the authors tried to introduce the immunochemical methods for determination of T-2 toxin. The enzyme-linked immunosorbent assay method using monoclonal antibody for T-2 toxin was applied to analyse T-2 toxin. The detection limit of T-2 toxin by ELISA method was 0.1 ppb. The correlation between ELISA and GC-MS method on these samples was very high. ELISA method developed for the detection and quantification of T -2 toxin in this paper possesses simplicity, high sensitivity and specific for T-2 toxin. Furthermore, the ELISA method with T-2 toxin monoclonal antibody was an excellent tool for the screening of Fusarium spp. which was suspected to produce T-2 toxin.

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Screening of Antioxidative Activity of Hot-Water Extracts from Medicinal Plants (한약재 열수추출물의 항산화효과 검정)

  • Kang, Mi-Young;Nam, Seok-Hyun
    • Applied Biological Chemistry
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    • v.43 no.2
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    • pp.141-147
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    • 2000
  • Interrelation between the antioxidative activity of hot-water extracts of 130 medicinal plants and their cellular antimutagenic activity was investigated. Antioxidative activity was evaluated by assaying electron-donation to DPPH free radical and scavenging of hydroxyl radical $({\cdot}OH)$ generated through Fenton rection, respectively. All medicinal plants examined in this study exhibited markedly electron-donating ability and radical scavenging ability in each assay system. The results demonstrated the fact that Pilbal (Piper longum L.) is the strongest in electron-donating activity, on the other hand, that Seokgok (Dendrobium moniliforme L.) is the strongest in ${\cdot}OH$ scavenging activity. When evaluated their antioxidative activities, 24 medicinal plants including Jimo (Anemarrhena asphodeloides Bunge) were found to be the medicinal plants carrying strong antioxidative activity, which exhibited more than 50% activity compared to the control group in both electron-donating and free radical scavenging. The experiment was also performed to examine whether 11 medicinal plants having significant antimutagenicity damage DNA in the presence of $Cu^{2+}$, showing the fact that all samples tested, except Taeksa (Alisma canaliculatum All. Br.), Paekjain (Nitraria sibirica Pall) and Ohyak (Lindera strychifolia Sieb. et Zucc. Villar) are capable of inducing DNA strand break. We also found that Taeksa and Paekjain strongly block DNA strand break induced by chemical mutagen mitomycin C.

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Safety Evaluation of Tobacco Substitute (Herbrette); Inhalation Toxicity, Mutagenicity and Immunotoxicity

  • Song, Kyung Seuk;Park, Kun Ho;Yoo, Gi Yong;Song, Sung-Ok;Kim, Hyun Woo;Kim, Jun Sung;Park, Jin Hong;Eu, Guk Joung;Hua, Jin;Cho, Hyun Sun;Hwang, Soon Kyung;Chang, Seung Hee;Tehrani, Arash Minai;Yu, KyeongNam;Chae, Chan Hee;Cho, Myung Haing
    • Toxicological Research
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    • v.20 no.4
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    • pp.365-374
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    • 2004
  • Inhalation toxicity, mutagenicity, and immunotoxicity tests were performed using a smoke generation system to investigate the safety of Herbrette, a tobacco substitute made with the leaves of Perilla frutescens. ICR mice were exposed to nicotine-free Herbrette smoke with concentrations of 0 (control), 4.08 $\pm$ 1.32 mg/$m^3$ (low dose), 7.72 $\pm$ 2.14 mg/$m^3$ (medium dose) and 12.83 $\pm$ 1.69 mg/$m^3$ (high dose) total particulate matters (TPM) for 4 weeks. When compared to the control group, the body weights, organ weights in the exposed groups did not show any significant differences. However, certain change of several serum chemical data and biochemical parameters were observed, however, the changes were within normal physiological ranges. Moreover, no changes in organ weight, and no gross/microscopic changes were observed between the exposed and control groups. Salmonella typhimurium reverse mutation, in vivo chromosomal aberration and micronucleus assays revealed that Herbrette did not induce mutagenicity. Upon evaluation of peripheral cellular immunity of mice through in vitro lymphocyte proliferation assay, no significant difference was observed in mean stimulation index between the exposed and control groups. Taken together, our results strongly suggest that Herbrette may not cause toxicity on mice under current condition.

Responses of Tobacco Photomixotrophic Cultured Cells to Various Herbicides (다양한 제초제에 대한 담배 Photomixotrophic 배양세포의 반응)

  • 권혜경;권석윤;이행순;윤의수;김진석;조광연;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.183-187
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    • 1999
  • To establish an efficient screening system for new herbicides using plant cultured cells, responses of tobacco photomixotrophic cultured (PH) cells to various herbicides with different modes of action were surveyed by measuring the cell growth and ion conductivity in medium. The cells were cultured in Murashige and Skoog (MS) medium containing 0.7mg/L 2,4-D, 0.3mg/L kinetin and 30 g/L sucrose at $25^{\circ}C$ in the light (100 rpm). Chemicals were treated to suspension cultures of tobacco PH cells at the time of subculture. The cell growth and ion conductivity in the medium were investigated on 12 days after chemical treatment. The ion conductivity assay gave well correlated results to the cell growth inhibition data. The responses of tobacco PM cells were dependent on the modes of action of chemicals tested. Atrazine, an inhibitor of photosynthetic electron transport (PET), strongly inhibited both the cell membrane and cell growth ($IC_{50}$/, about 1 $\mu$M). Butachlor (an inhibitor of cell division), glufosinate (an inhibitor of amino acid biosynthesis), and fluridone (an inhibitor of carotenoid biosynthesis) showed a dose-dependent inhibition. However, Quinclorac, a herbicide with an auxin activity, did not affect the cell growth and ion leakage. These results suggested that tobacco PM cells is suitable materials for the simple screening of new herbicides such as PET, amino acid biosynthesis, ceil division inhibitors by measuring the cell growth and ion conductivity.

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