• Title/Summary/Keyword: cell fractions

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Studies on the Chemical Constituents Biological Activities of Mulberry Extracts (오디(Mulberry) 추출물의 성분분석 및 생리활성에 관한 연구)

  • Lee, Sang-Keug;Lee, Hak-Ju;Kang, Ha-Youg;Choi, Don-Ha;Jo, Hyun-Jin;Lee, Tae-Seong
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.1
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    • pp.105-113
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    • 2009
  • The freezed Mulberry (10 kg) was extracted with 80% EtOH, concentrated, and fractionated with a series of n-hexane, ethyl acetate, and water on a separatory funnel. A portion of ethyl acetate soluble (22 g) was chromatographed on a Sephadex LH-20 column using a series of aqueous methanol as eluents. The isolated compounds were identified by cellulose TLC, $^1H-$,$^{13}C$-NMR, FAB, and EI-MS. Quercetin-3-O-${\beta}$-D-glucopyranoside (Compound I), protocatechuic acid (Compound II), p-hydroxybenzoic acid (Compound III) were isolated from the ethyl acetate soluble fraction. In antioxidative activities of the fractionated extractives using DPPH radical scavenging test, EtOAc and water soluble fractions indicated better than BHT as contro and in in vitro tests using MTT assay, there was no cytotoxicity. Also, tyrosinase inhibition and anticancer activities were not so good, but there may be a potential as a cosmetic raw material because the cell extension effect was excellent.

Cellular responses and proteomic analysis of hemolytic Bacillus cereus MH-2 exposed to epigallocatechin gallate (EGCG) (Epigallocatechin Gallate (EGCG)에 노출된 용혈성 Bacillus cereus MH-2의 세포 반응 및 프로테옴 분석)

  • Kim, Dong-Min;Park, Sang-Kook;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.260-268
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    • 2016
  • The aim of this work was to investigate the cellular responses and proteomic analysis of Bacillus cereus MH-2 exposed to EGCG. Strain MH-2 was isolated from commercial Ssamjang and has the hemolytic activity. Survival of the MH-2 strain with time in the presence of different concentrations of EGCG under sublethal conditions was monitored. The amount of alginate from MH-2 strain decreased depending on the increasing concentrations of EGCG and increased depending on the exposure time at any particular EGCG concentration. Analysis of SDS-PAGE and Western blot using anti-DnaK and anti-GroEL revealed that two stress shock proteins, 70 kDa DnaK and 60 kDa GroEL were found to decrease in proportion to the EGCG concentration in exponentially growing cultures. Scanning electron microscopic analysis demonstrated the presence of protrusions and fused rod forms on the cells treated with EGCG. 2-DE of soluble protein fractions from MH-2 cultures showed 20 protein spots changed by EGCG exposure. These proteins involved in enterotoxins (hemolysin BL lytic component L1 and hemolysin BL-binding protein), chaperons (DnaK and GroEL), cell defense (peptidase M4 family proteins), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. These results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on B. cereus MH-2.

Identification and quantification of major malonyl ginsenosides isolated from Panax ginseng C.A. Meyer (인삼(Panax ginseng C.A. Meyer)로부터 Malonyl ginsenoside의 분리 및 정량분석)

  • Shin, Woo Cheol;Jung, Jiyun;Na, Hyeon Seon;Bo, Jeon Hwang;Kim, Hyoung-Geun;Yoon, Dahye;Choi, Bo-Ram;Lee, Young-Seob;Kim, Geum-Soog;Baek, Nam-In;Lee, Yi;Lee, Dae Young
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.375-384
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    • 2019
  • The root of Panax ginseng C.A. Meyer were extracted with 70% aqueous EtOH and the concentrates were partitioned into MeOH and H2O fractions using Diaion HP-20. The repeated SiO2 or octadecyl SiO2 column, and MPLC for the MeOH fraction led to isolation of four malonyl ginsenosides. The chemical structures of these compounds were determined as malonyl ginsenoside Rd (1) malonyl ginsenoside Rc (2) malonyl ginsenoside Rb2 (3) malonyl ginsenoside Rb1 (4) based on spectroscopic analyses including Nuclear magnetic resonance and HR-TOF/MS. The contents of malonyl ginsenoside Rb1 was highist as 5.44 mg/g of five years of ginseng. And malonyl ginsenoside Rd was lowest as 0.11 mg/g of six years of ginseng. Additionally, the malonyl ginsenoside Rd exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line.

Isolation and quantitative analysis of metabolites from Scrophularia buergeriana and their hepatoprotective effects against HepG2 Cells (현삼 (Scrophularia buergeriana)에서 분리한 화합물의 함량분석 및 간세포 보호 효과)

  • Na, Hyeon Seon;Oh, Seon Min;Shin, Woo Cheol;Bo, Jeon Hwang;Kim, Hyoung-Geun;Yoon, Dahye;Yang, Seung Hwan;Lee, Young-Seob;Kim, Geum-Soog;Baek, Nam-In;Lee, Moon-Soon;Lee, Dae Young
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.399-406
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    • 2019
  • The roots of Scrophularia buergeriana were extracted with 80% aqueous Methanol and the concentrates were partitioned into EtOAc, n-BuOH, and H2O fractions. The repeated silica gel or octadecyl SiO2column, and medium pressure liquid chromatographies for the n-BuOH fraction led to isolation of phenylethanoid glycosides and iridoid glycosides. The chemical structures of these compounds were determined as harpagoside (1), angoroside C (2), aucubin (3) and acetoside (4) based on spectroscopic analyses including nuclear magnetic resonance and MS. A simple and efficient HPLC with UV detection method for the simultaneous determination of the four compounds (1-4) has been developed and applied to their content determination in the S. buergeriana. The roots were extracted by 80% methanol, and the contents of 1, 2, 3, and 4 were determined to 11.5, 7.6, 41.2, and 4.8 mg/g, respectively. Additionally, angoroside C (2) and acetoside (4) exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line.

Conformal Radiotherapy in a Patient with Cancer at the base of the Tongue in a Previously Irradiated Area (방사선치료 조사영역 내에 발생한 설암 환자에서 입체조형방사선치료 경험 : 증례보고)

  • Cho Moon-June;Kim Ki-Hwan;Kim Byung-Kook;Song Chang-Joon;Kim Jun-Sang;Kim Jae-Sung;Jang Ji-Young
    • Korean Journal of Head & Neck Oncology
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    • v.17 no.1
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    • pp.59-62
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    • 2001
  • Objectives: We report an interim result of conformal radiotherapy in a patient with early stage cancer at the base of the tongue, which developed in a previously irradiated area. Materials and Methods: A 64-year-old male patient was diagnosed with T4N0M0 supraglottic cancer. He received 72Gy of radiation therapy from 21 November 1988 to 24 February 1989. He had local failure and underwent a salvage total laryngectomy on 28 August 1989. Subsequently, he did well. In early 1999, he suffered from throat pain. He had a 2.5cm ulcerative mass at the base of his tongue, in the area that had been irradiated previously. Biopsy showed squamous cell carcinoma. After workup, he was diagnosed with base of tongue cancer with T2N0M0. Surgery was not feasible because the morbidity was not acceptable. Since it was difficult to re-irradiate the area with a curable dose using conventional 2D radiation therapy with an acceptable morbidity, we decided to try conformal radiotherapy. We used 7 static beam ports with field sizes from $7x6.4\;to\;8x8cm^2$, using 6 and 10MV photons. The fractionation regimen was 1.8Gy, 5 times per week. He received 64.8Gy in 36 fractions from 9 April 1999 to 1 June 1999. Results: In the 21 months since radiotherapy, the patient has not experienced any acute or chronic complications, such as xerostomia. He experienced relief of pain shortly after the start of radiotherapy, showed a complete response, and is still doing well. Conclusion: Conformal radiotherapy can be used to treat cancer that develops within a previously irradiated field, with curative intent.

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The Inhibitory Effect of Grapefruit Seed Extracts on the Physiological Function of Enterobacter pyrinus (Grapefruit 종자추출물이 Enterobacter pyrinus의 생리기능에 미치는 영향)

  • Lee, Tae-Ho;Jeong, Sook-Jung;Lee, Sang-Yeol;Kim, Jae-Won;Cho, Sung-Hwan
    • Korean Journal of Food Science and Technology
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    • v.27 no.6
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    • pp.985-990
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    • 1995
  • Grapefruit seed extracts(GFSE) have some unknown compounds which exhibit the antibiotic activities aganist microorganisms including bacteria and fungi. We have examined the effects of GFSE on the growth of Enterobacter pyrinus which was isolated from necrotic lesions of pear trees. During the cultivation, the growth of the bacteria was strongly inhibited at the low concentration(0.01%, w/w) of GFSE. Hydrophobic fraction extracted from GFSE by mixed solvents (chloroform : methanol : water, 1 : 2 : 0.8, v/v/v) had components which inhibited the growth of bacteria. There was, however, no inhibitory effect of GFSE on the activities of several enzymes including hexokinase, glucose 6-phosphate dehydrogenase, malate dehydrogenase and succinate dehydrogenase. $O-nitrophenyl-{\beta}-D-galactopyranoside(ONPG)$, the artificial substrate of ${\beta}-galactosidase$ was hydrolyzed in the presence of GFSE, indicating that the membrane was pertubated by the GFSE. From the results it was suggested that the antibiotic activity of GFSE is due to the change of membrane permeability of cell. GFSE was fractionated by high performance liquid chromatography equipped with $C_{18}$ reverse phase column. Among active fractions, three peaks were identified as 1-chloro-2-methyl-benzene (o-toluene), N,N-dimethyl-benzenemethaneamine, 1-[2-(2-ethylethoxy)ethoxy]-4- (1,1,3,3-tetramethyl)-bezene, respectively, while the other three remained unidentified.

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Antitumoral, Antioxidant and Antimicrobial Activities of Solvent Ftactions from Grifola umbllatus (저령추출물의 항암, 항산화 및 항균효과)

  • 하영득
    • Food Science and Preservation
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    • v.8 no.4
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    • pp.481-487
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    • 2001
  • Grifola umbellatus was extracted using methanol, and the extract was further fractionated by water and ethyl acetate. Assay of each fraction with MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium-bromide] revealed significant cytotoxicity effect of the methanol extract of Grifola umbellatus against human gastric cancer cell but not normal human lymphocytes. The methanol extract showed the highest antioxidant activity as well. Antimicrobial activity of Grifola umbellants against Helicobacter pylori was higher in method extract than in other fractions. Grifola umbellatus had a significant inhibitory effect on Helicobacter pylori reducing both its growth and urease activity. These results show that the methanol extract of Grifola umbellatus possesses therapeutic potential on gastric diseases.

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Inhibitory Effect of Triticum aestivum Ethanol Extract on Lipid Accumulation in 3T3-L1 Preadipocytes (3T3-L1 세포에서 소맥엽 에탄올추출물의 지질생성 억제효과)

  • Lee, Sun-Hee;Xin, Mingjie;Luyen, Bui Thi Thuy;Cha, Ji-Yun;Im, Ji-Young;Kwon, Se-Uk;Lim, Sung-Won;Suh, Joo-Won;Kim, Young-Ho;Kim, Dae-Ki;Lee, Young-Mi
    • YAKHAK HOEJI
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    • v.55 no.6
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    • pp.478-484
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    • 2011
  • Non-alcoholic fatty liver disease is known to be frequently associated with obesity and type 2 diabetes. We examined the effects of EtOH extracts from Triticum aestivum on lipid accumulation during the differentiation of 3T3-L1 preadipocytes to screening the candidate materials in preventing non-alcoholic fatty liver disease. The lipid level in adipocytes was determined by Oil Red O staining. The treatment of 50% ethanol, but not water and 100% ethanol extracts, from Triticum aestivum at concentration of 0.5 $mg/ml$ inhibited lipid accumulation in 3T3-L1 cells, revealing no cell toxicity. Thus, the fractions of $CH_2Cl_2$, EtOAc and BuOH were separated from 50% EtOH extract to characterize anti-adipogenic effect. The $CH_2Cl_2$ fraction at concentration of $50{\mu}g/ml$ effectively inhibited the lipid accumulation in the adipocytes compared to those of EtOAc and BuOH at concentration of $50{\mu}g/ml$. The intracellular triglyceride accumulation also was significantly reduced by treatment of $CH_2Cl_2$ fraction in concentration-dependent manner. Western blot analysis showed that the $CH_2Cl_2$ fraction attenuated the intracelluar level of fatty acid synthase(FAS) accompanied by attenuated expression of Peroxidase proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$) adipogenic transcription factor. These results suggest that $CH_2Cl_2$ fraction from 50% EtOH extract of Triticum aestivum may has the potent anti-adipogenic effects by inhibiting the transactivation of $PPAR{\gamma}$.

A Potent Tissue Destructive Activity of Secreted Proteins of Aeromonas hydrophila (조직 괴사 활성을 지닌 Aeromonas hydrophila 의 분비 단백질에 관한 연구)

  • Kim, Kyu Lee;Choe, Yunjeong;Kang, Ho Young
    • Journal of Life Science
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    • v.25 no.2
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    • pp.214-222
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    • 2015
  • Aeromonas hydrophila is the most common water fish pathogen and cause diseases such as hemorrhagic septicemia, dropsy, ulceration and asymptomatic septicemia. A. hydrophila secretes many extracellular products (ECPs) which contribute to effective infection, wide distribution and great adaptability to environmental changes. Crude ECPs of A. hydrophila CK257, a strain used in this study, exhibits a toxic activity to the animals including mouse, rabbit and fish. Toxic symptoms were indicated by tissue damage and skin injuries in animal. When ECPs were subcutaneously injected to animals, skin damages were observed, appearing like necrosis. Preliminary research demonstrated that the active factors are protein component. The crude ECPs were collected after ammonium sulfate precipitation of cell-free culture supernatant. ECPs were fractionated with the use gel filtration chromatography. Five ECP fractions were obtained, of which one fraction was found to be toxic to goldfish. MALDI-TOF analyses provided two interesting proteases called M35 and M28. Both M35 and M28 are known as metalloprotease. Accordingly, proteins in an active fraction exhibited caseinolytic activity. These proteins were difference of caseinolytic activity under different metallic ions. Also active fraction has elastolytic activity. These results suggested that peptidase M28 and M35 may be a candidate factor for tissue necrosis activity about infection with A. hydrophila.

Potential Effect of Solvent Fractions of Taraxacum mongolicum H. on Protection of Gastric Mucosa (민들레(Taraxacum mongolicum H.) 추출분획물이 위장보호에 미치는 효능 평가)

  • Han, So-Heui;Hwang, Jung-Keun;Park, Soo-Nam;Lee, Kil-Hong;Ko, Kang-Il;Kim, Ki-Su;Kim, Ki-Ho
    • Korean Journal of Food Science and Technology
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    • v.37 no.1
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    • pp.84-89
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    • 2005
  • Mongolian dandelion (Taraxacum mongolicum H.) extracted with solvents having different polarities were prepared to investigate protection activity of gastric mucosa. Ethanol extracts were successively reextracted with methylene chloride, and n-butanol. Concentrations of total flavonoids and luteolin in n-butanol extracts were 27.75 and 1.14%, respectively, much higher than those of other solvent extracts. Results of anti-microbial acitivity test against Helicobacter pylori and urease inhibition test revealed n-butanol extract exerted higher inhibition (13.16%) than other solvent extracts. Based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, n-butanol extract showed the highest efficiency of free radical-scavenger activity among the extracts ($SC_{50}\;47{\mu}g/mL$). Erythrocytic photohemolysis tests, for the protection of cell membrane showed that half-life of human erythrocytes was increased by the addition of n-butanol extract $({\tau}_{50}=172min;C=5.0{\mu}g/mL)$. These results indicate n-butanol extract of Mongolian dandelion may be useful as a adjuvant for gastric diseases.