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Isolation of Alliin in Garlic and Its Quantitative Determination by High Performance Liquid Chromatography and Studies on the Antimicrobial Efforts of Alliin and Ethanol Extracts from Korean Garlic(Alliium sativum L.) (마늘 중 고속 액체 크로마토그래피에 의한 알린의 분리 및 정량과 Alliin과 에탄올 추출물의 항균효과에 관한 연구)

  • 위성언
    • The Korean Journal of Food And Nutrition
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    • v.16 no.4
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    • pp.296-302
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    • 2003
  • First. the purification and analysis of alliin in garlic from different origins by alliin-HPLC determination method were studied. Allinase in garlic was inactivated by heating in boiling water followed by extraction of alliin in garlic with 80% methanol. To remove free amino acids and alliin homologs in garlic, garlic extract was separated by cation exchange column which was packed with amberlite CG-120 resin using 40L d-water as eluent. Alliin in garlic extract was crystallized in a mixture of acetone (50$^{\circ}C$):H$_2$O:acetic acid=70:29:1 and then recrystallized in a mixture of acetone (50$^{\circ}C$):H$_2$O:acetic acid=75:24:1. Obtained alliin was identified by melting point. TLC, microscope observation and mass spectrometry. High performance liquid chromatography (HPLC) following pre-column derivatization of cystein derivatives with o-phthaldialdehyde/2-mercaptoethanol has succeessfully been applied to the analysis of various garlics. Each alliic of standard solution and garlic extract was derivatized to isoindole derivative by o-phthaldialdehyde /2-mercaptoethanol and then analyzed by HPLC. Six point calibration was done by using alliin peak area. Lineality was observed at 0 ∼ 1.0mg/ml of alliin concentration. Weighted regression line function was Y=6254X - 256077. By this function, alliin contents in various garlics were 0.34 ∼ 0.73% fresh weight. Second study was designed to evaluate the effects of garlic extracts of various concentrations on the growth of various pathogenes (Eubacterium limonsum, Bacteroides fragilis, Salmonella typhimurium, Salmonella typhi, Shigella sonnei, Kiebsiella pneumoniae, Enterobacter cloacae, Pserdomonas aeruginosa, Escherichia coli). For antimicrobial effects against microorganism, totally minimal inhibition concentrations (MIC) of alliin were from 5,000 to 20,000ppm. MIC of ethanol extract were 1,250 to 10,000ppm.

Estimation of Impurities from Commercially Available Glycyrrhizin Standards by the HPLC/ESI-MS (HPLC/ESI-MS에 의한 글리시리진 표준품의 불순물 추정)

  • Myung, Seung-Woon;Min, Hye-Ki;Kim, Myungsoo;Kim, Young Lim;Park, Seong-Soo;Cho, Jung Hee;Lee, Jong-Chul;Cho, Hyun-Woo;Kim, Taek-Jae
    • Analytical Science and Technology
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    • v.13 no.4
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    • pp.504-510
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    • 2000
  • The impurity profiles from the raw materials of glycyrrhizin were performed by the high performance liquid chromatography (HPLC)/electrospray ionization (ESI)- mass spectrometry (MS). For the HPLC experiment, a $C_{18}$($3.9{\times}300mm$, $10{\mu}m$) column was used and the mobile phase was acetic acid/$H_2O$ (1:10):acetonitrile=3:2 with a flow rate of 0.8 ml/min. The effluent was splitted into the ratio of 50:1 and went into the ESI-MS. Three to six impurities were found and informed of the identification of the structure of the impurities by ESI-MS. And the structures of impurities were suggested to a hydroxy-glycyrrhizin which is added with hydroxy group (-OH) in the glycyrrhetic acid moiety and a reduced-glycyrrhizin which the position of 12 of the glycyrrhetic acid moiety is reduced. The purities of the standard materials were about 90%.

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Assessment of three European fuel performance codes against the SUPERFACT-1 fast reactor irradiation experiment

  • Luzzi, L.;Barani, T.;Boer, B.;Cognini, L.;Nevo, A. Del;Lainet, M.;Lemehov, S.;Magni, A.;Marelle, V.;Michel, B.;Pizzocri, D.;Schubert, A.;Uffelen, P. Van;Bertolus, M.
    • Nuclear Engineering and Technology
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    • v.53 no.10
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    • pp.3367-3378
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    • 2021
  • The design phase and safety assessment of Generation IV liquid metal-cooled fast reactors calls for the improvement of fuel pin performance codes, in particular the enhancement of their predictive capabilities towards uranium-plutonium mixed oxide fuels and stainless-steel cladding under irradiation in fast reactor environments. To this end, the current capabilities of fuel performance codes must be critically assessed against experimental data from available irradiation experiments. This work is devoted to the assessment of three European fuel performance codes, namely GERMINAL, MACROS and TRANSURANUS, against the irradiation of two fuel pins selected from the SUPERFACT-1 experimental campaign. The pins are characterized by a low enrichment (~ 2 wt.%) of minor actinides (neptunium and americium) in the fuel, and by plutonium content and cladding material in line with design choices envisaged for liquid metal-cooled Generation IV reactor fuels. The predictions of the codes are compared to several experimental measurements, allowing the identification of the current code capabilities in predicting fuel restructuring, cladding deformation, redistribution of actinides and volatile fission products. The integral assessment against experimental data is complemented by a code-to-code benchmark focused on the evolution of quantities of engineering interest over time. The benchmark analysis points out the differences in the code predictions of fuel central temperature, fuel-cladding gap width, cladding outer radius, pin internal pressure and fission gas release and suggests potential modelling development paths towards an improved description of the fuel pin behaviour in fast reactor irradiation conditions.

Purification of TGF-$\beta$ 1 from Human Platelets by an Improved Method (개량된 방법에 의한 사람혈소판으로부터 TGF-$\beta$ 1의 분리)

  • 신충건;김상국;문병조;김평현;전계택;남상욱;김장환;이종원
    • KSBB Journal
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    • v.14 no.1
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    • pp.9-16
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    • 1999
  • Transforming growth factor $\beta$1(TGF-$\beta$1) has potentials to be used as a new therapeutic agent. However, studies with TGF-$\beta$ were hindered by its high cost. In this study, we developed an improved method to purify TGF-$\beta$1 from human platelets, for which four purification steps were used: platelet extraction, gel filtration, cation exchange chromatography, and reverse phase high performance liquid chromatography. After a final step of purification, a pure protein with a molecular weight of 25,000 corresponding to the commercially available TGF-$\beta$1 was obtained, which were confirmed by silver staining and Western blotting after SDS-polyacrylamide gel electrophoresis (SDS-PAGE). It was confirmed by the inhibitory effects of TGF-$\beta$1 on a mink lung epithelial cell line that the purified TGF-$\beta$1 had its biological activity, whose activity is slightly higher than that of the commercially available TGF-$\beta$1. About 3.7$\mug of the purified TGF-$\beta$1 was obtained from 10 units of concentrated human platelets, the final yield of which is about 21%.

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Isolation and Characterization of MMP-1 Inhibitor Peptide from Crataegus pinnatifida Bunge in Fibroblast Cell Line HS68 Cells (아가위(Crataegus pinnatifida Bunge)로 부터 HS 68세포의 MMP-1에 대한 저해활성 물질의 분리)

  • Lee, Se-Young;Chun, Hyug;Cho, Hong-Yun;An, Jeung-Hee
    • Applied Biological Chemistry
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    • v.46 no.1
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    • pp.60-65
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    • 2003
  • MMP-1 inhibitory compounds were isolated from 120 Korean traditional edible plants. UP- 1 activity significantly increased linearly with increasing UVB dose in normal human foreskin fibroblast HS68 cell, showing maximum activity at approximately 35 $mJ/cm^2$, whereas in HaCaT cell, normal human keratinocyte, no increase was observed. Maximum secretion of MMP-1 after UVB treatment occurred around 36-48 k after treatment. MMP-1 inhibitory compound isolated from cold-water fraction of Cataegus pinnatifida Bunge showed the mort potent activity. The MMP-1 inhibitory compound was deduced as a peptide based on the fact that pronase digestion decreased the activity whereas periodate oxidation did not. The most potent UP- 1-inhibitory protein, CP-2Va-2, showing an activity of 88.5% against MMP-1, was isolated through sequential column chromatography on DEAE-Toyopearl 650C, Butyl-Toyopearl 650M, and Bio-Gel P-30. Molecular weight of CP-2Va-2 determined through high performance liquid chromatography and SDS PACE was 19 and 20 kDa. respectively, signifying a monomeric structure.

Antioxidant Activity of Ripe Fruit Components of Rubus coreanus: Extraction Using Porous Polymer Resins (다공성 합성흡착제를 이용한 복분자 딸기(Rubus coreanus) 농축액의 항산화 효과)

  • Choi, Se-Jin;Lee, Yeon-Sil;Kim, Jin-Kyu;Chung, Cha-Kwon;Kang, Il-Jun;Lim, Soon-Sung
    • Korean Journal of Food Science and Technology
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    • v.43 no.2
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    • pp.149-155
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    • 2011
  • This study was carried out to develop a method of removing unnecessary sugars and high-molecular weight soluble components from water extract by using various highly porous polymer resins: these resins are widely used as adsorbents for polyphenolic compounds. Each anthocyanin-rich fraction (ARF) that was eluted from column packed with 4 different resins (SEPABEADS SP207, 700, 850 and Diaion HP 20) was obtained from an aqueous extract of the ripe fruits of Rubus coreanus, which is well-known in Korea as "Bok-bun-ja". Among the above mentioned resins, Diaion HP20 had the highest efficacy and provided maximum yield of the ARF. The ARF eluted from the column packed with HP 20 exhibited the strongest antioxidant activity in vitro. Bioactive phytochemicals extracted from Rubus coreanus were investigated using the on-line HPLC-$ABTS^{{\cdot}+}$ system, and the resulting 6 peaks were identified as radical-scavenging components. By using liquid chromatography-mass spectrometry, 5 peaks were obtained, and these were identified as cyanidin-3-sambubioside, cyanidin-3-glucoside, cyanidin-3-xylosylrutinoside, cyanidin-3-rutinoside and pelargonidin-3-rutinoside. These results indicate that the use of appropriate porous resin (Diaion HP 20) leads to an increase in the yield of bioactive components and enhancement of their biological properties.

Liquid Culture of Basidiomycetes on Natural Media (천연물을 이용한 담자균의 균사체 배양)

  • Kim, Seon-Hee;Lee, Jong-Suk;Park, Kyung-Sook;Lee, Jae-Sung;Lee, Hang-Woo;Park, Shin
    • The Korean Journal of Mycology
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    • v.27 no.6 s.93
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    • pp.373-377
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    • 1999
  • This study was designed to develop functional drink from jujube extract through a simple submerged culture of three basidiomycetes species. The optimum Brix and pH of the jujube extract for culturing the Ganoderma lucidum appeared to be 5 Brix and pH 4. Ten days of culture period produced maximum mycelium. The optimum Brix and pH of the jujube extract for culturing the Coriolus versicolor appeared to be 5 Brix and pH 5. Ten days of culture period produced maximum mycelium. The optimum Brix and pH of the jujube extract for culturing the Phellinus igniarius appeared to be 3 Brix and pH 5. For the maximum mycelial production eighteen days of culture period was required for Phellinus igniarius. The antitumor activity of the polysaccharides extracted from the fermented drinks was demonstrated through the tumor cell line experiments. The $IC_{50}$ values of the jujube drinks fermented with Ganoderma lucidum and Phellinus igniarius against stomach cancer cell line appeared to be one fourth that of the jujube drink which was not fermented with basidiomycetes.

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A 3D "In Vitro" Model to Study Hyaluronan Effect in Nasal Epithelial Cell Line Exposed to Double-Stranded RNA Poly(I:C)

  • Albano, Giusy Daniela;Bonanno, Anna;Giacomazza, Daniela;Cavalieri, Luca;Sammarco, Martina;Ingrassia, Eleonora;Gagliardo, Rosalia;Riccobono, Loredana;Moscato, Monica;Anzalone, Giulia;Montalbano, Angela Marina;Profita, Mirella
    • Biomolecules & Therapeutics
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    • v.28 no.3
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    • pp.272-281
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    • 2020
  • Environmental agents, including viral and bacterial infectious agents, are involved in the alteration of physicochemical and biological parameters in the nasal epithelium. Hyaluronan (HA) has an important role in the regulation of tissue healing properties. High molecular weight HA (HMW-HA) shows greater anti-inflammatory responses than medium molecular weight HA (MMW-HA) and low molecular weight HA (LMW-HA). We investigated the effect of HMW-HA, MMW-HA and LMW-HA on the regulation of physicochemical and biological parameters in an "in vitro" model that might mimic viral infections of the nasal epithelium. Human nasal epithelial cell line RPMI2650 was stimulated with double-stranded RNA (dsRNA) Poly(I:C) for 5 days in air-liquid-interface (ALI) culture (3D model of airway tissue). dsRNA Poly(I:C) treatment significantly decreased transepithelial electrical resistance (TEER) in the stratified nasal epithelium of RPMI2650 and increased pH values, rheological parameters (elastic G' and viscous G''), and Muc5AC and Muc5B production in the apical wash of ALI culture of RPMI2650 in comparison to untreated cells. RPMI2650 treated with dsRNA Poly(I:C) in the presence of HMW-HA showed lower pH values, Muc5AC and Muc5B production, and rheological parameters, as well as increased TEER values in ALI culture, compared to cells treated with Poly(I:C) alone or pretreated with LMW-HA and MMW-HA. Our 3D "in vitro" model of epithelium suggests that HMW-HA might be a coadjuvant in the pharmacological treatment of viral infections, allowing for the control of some physicochemical and biological properties affecting the epithelial barrier of the nose during infection.

Scutellaria Extract Decreases the Proportion of Side Population Cells in a Myeloma Cell Line by Down-regulating the Expression of ABCG2 Protein

  • Lin, Mei-Gui;Liu, Li-Ping;Li, Chen-Yin;Zhang, Meng;Chen, Yuling;Qin, Jian;Gu, Yue-Yu;Li, Zhi;Wu, Xin-Lin;Mo, Sui-Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7179-7186
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    • 2013
  • Background and Aims: Scutellaria is one of the most popular traditional Chinese herbal remedies against various human diseases, including cancer. In this study, we examined the active effects of Scutellaria extract and its main flavonoid constituents on the proportion of side population cells within human multiple myeloma cell line RPMI8226 in vitro and explored the potential molecular mechanisms involved. Materials and Methods: The contents of flavonoids in ethanolic extract of Scutellaria baicalensis Georgi were determined using high performance liquid chromatography. The antiproliferative effect of the ethanolic extract on RPMI-8226 was determined by CCK assay. Apoptosis was measured by annexin combining with propidium iodide in a flow cytometer. Cell cycle analysis was performed by propidium iodide staining in combination with flow cytometry analysis. Hoechst 33342 exclusion assay was used for the identification of side population within RPMI8226 cells. The expression of ABCG2 protein was assessed by Western blotting assay. Results: The content of major flavonoids constitutents of Scutellaria extract was baicalin (10.2%), wogonoside (2.50%), baicalein (2.29%), and wogonin (0.99%), respectively. The crude Scutellaria extract did not show significant anti-proliferative effect, apoptosis induction and cell cycle arrest in RPMI-8226 within the concentrations of $1-75{\mu}g/mL$. However, the ethanolic extract, baicalein, wogonin and baicalin reduced the side population cells in RPMI-8226, and data showed that baicalein and wogonin had stronger inhibitory effects. Correspondingly, they also exhibited significant effects on decreasing the expression level of ABCG2 protein in RPMI-8226 in vitro. Conclusions: Our results for the first time demonstrated a novel mechanism of action for Scutellaria extract and its main active flavonoids, namely targeting SP cells by modulating the expression of ABCG2 protein. This study provides an insight for new therapeutic strategies targeting cancer stem cells of multiple myeloma.

A Study of Sterilization Effect of Long-wavelength UVA-LED Irradiation on Bacteria Causing Eye Diseases (장파장의 자외선 LED 광원을 이용한 안질환 세균의 살균효과)

  • Lee, Cheol-Woo;Jeong, Kyeong-In;Hwang, Kwang-Ha;Lee, Seok-Ju;Yoo, Geun-Chang
    • Journal of Korean Ophthalmic Optics Society
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    • v.17 no.1
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    • pp.99-105
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    • 2012
  • Purpose: The purpose of this study is to demonstrate inactivation effect of UVA-LED ultraviolet radiation upon Pseudomonas aeruginosa and Staphylococcus aureus which are the major bacteria causing eye diseases. Methods: The small sterilization device was made using UVA-LED of 400 nm. After Pseudomonas aeruginosa was diluted to $10^{-7}$ and Staphylococcus aureus to $10^{-5}$ and diluted solutions were put onto each liquid medium. They were irradiated by 400 nm of UVA for different amount of time; 15 min, 30 min, 60 min, 120 min, 240 min, 360 min and 480 min each. Results: The data from sterilization test was solved to regression line equation and the target log inactivation was obtained. The 3 log inactivation UV irradiation value of Pseudomonas aeruginosa was 54,847 UV dose ($mJ/cm^2$) and irradiation time was 135.42 min while the 3 log inactivation of Staphylococcus aureus was 39,066 UV dose ($mJ/cm^2$) and irradiation time was 98.72 min. Conclusions: The inactivation effect of sterilization method using 400 nm of UVA-LED upon Pseudomonas aeruginosa and Staphylococcus aureus has been verified and it is considered as a useful method in inactivating the contact lenses.