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Isolation of Polyphenol Compounds from the Leaves of Korean Persimmon (Diospyrus kaki L. Folium) (한국산 감잎로부터 Polyphenol계 생리활성물질 분리)

  • An, Bong-Jeun;Bae, Man-Jong;Choi, Hee-Jin;Zhang, Yun-Bin;Sung, Tae-Soo;Choi, Cheong
    • Applied Biological Chemistry
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    • v.45 no.4
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    • pp.212-217
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    • 2002
  • We purified polyphenols from persimmon leaf and tested their biological activity. The 60% acetone extract was lyophilized and applied to test enzyme inhibition of glucosyltransferase and tyrosinase. GTase was 82.4% inhibited at $1.8{\times}10^{-1}$ mg/ml and tyrosinase 21.7% inhibited at 0.8 mg/ml. The acetone extract was fractionated into F-1, 2, 3, 4, 5 by Sephadex Q-50 gel filtration and the fraction-1 and 2 showed higher enzyme inhibition activity than the other fractions. To the Proteinase K treatment and autoclaving of the two fractions had no effect on the enzyme activity, but these results suggested that active fraction was not protein but phenol ring completed compounds. By Sephadex LH-20, MCI-gel and Bondapak $C_{18}$ column chromatographies, compouds 1, 2, 3 and 4 from F-1 fraction, compounds 5 and 6 from F-2 fraction and compounds 7 , 8 from F-3 fraction were purified and re-crystallized. The purified compounds was assumed to be condensed tannins of frame flavan-3-ol frame on the basis of color reagent reaction and to be a mixture of monomer, dimer and trimer according to TLC analysis.

Study on Optical Characteristics of 8-Hydroxyquinoline Synthesized Derivative as Sensing Material of the Fiber-Optic Copper Ion Sensor in Aqueous Environment (수질환경에서 광섬유 센서의 구리 이온 감지 물질로서 8-Hydroxyquinoline 합성유도체의 광학적 반응 특성 연구)

  • Kim, Beom Kyu;Park, Byung Gi
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.12
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    • pp.100-105
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    • 2017
  • An 8-hydroxyquinoline compound that was synthesized with 8-hydroxyquinoline-2-carboxaldehyde and 4-aminoantipyrine was investigated for use as the sensing material of a fiber-optic copper ion sensor in an aqueous environment. The experiment was conducted with a fiber-optic measurement system, in order to evaluate the relationship between the absorbance peak and copper ion concentration. The synthesized derivative exhibited a (highly selective) chromogenic phenomenon for copper ions among various metal ions in an aqueous environment and showed a specific absorbance peak at a wavelength of 530 nm for copper ions. The effect of mercury ions was investigated to evaluate the selectivity of the prepared synthesized derivatives toward Cu ions. The absorbance was measured at various concentration ratios of Cu and Hg ions (Cu:Hg ratios from 0.05 to 20), and it was found that the absorbance at 530 nm tended to increase with increasing Cu ion concentration. The experimental results also showed the linear relationship between the logarithmic concentration of copper ions and the specific absorbance peak at a wavelength of 530 nm. These results indicate that the synthesized 8-hydroxyquinoline compound has selectivity for copper ions and can be used as a sensing material for fiber-optic copper ion sensors.

Development of an Enzyme-Linked Immunosorbent Assay Method for Residual Host Cell derived Proteins in Recombinant Antibody Drug Production (재조합 항체의약품의 생산시 생산세포주 유래 단백질 검출을 위한 ELISA 방법 개발)

  • Joung, Chan-Hi;Lim, Sang-Min;Koo, Yoon-Mo;Lee, Yong-Yoon;Son, Young-Su;Kim, Hyun-Il;Park, Heung-Rok
    • KSBB Journal
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    • v.21 no.3
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    • pp.212-219
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    • 2006
  • The purpose of this study was to develop a assay system of host cell-derived residual proteins in final pharmaceutical products. Accurate and simple assay system for host cell-derived proteins(HCPs) is very important test item in pharmaceutical qualification control. In this study, methods for quantification of residual HCPs in recombinant anti-GPIIbIIIa antibody were developed using a process-specific immunoligand assay which was based on the Enzyme linked Immunosorbent assay(ELISA) system. The assay had a detection limit of 10.8 ng/ml of HCPs with a product concentration of 1 mg/ml. The practical implication of these results is that the developed ELISA system can be used for HCPs qualification control and this system will be applicable to develop another ELISA system of different antibody drug.

The development of anti-DR4 single-chain Fv (ScFv) antibody fused to Escherichia coli alkaline phosphatase (대장균의 alkaline phosphatase가 융합된 anti-DR4 single-chain Fv (ScFv) 항체의 개발)

  • Han, Seung Hee;Kim, Jin-Kyoo
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.10-17
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    • 2016
  • Enzyme immunoassay to analyze specific binding activity of antibody to antigen uses horseradish peroxidase (HRP) or alkaline phosphatase (AP). Chemical methods are usually used for coupling of these enzymes to antibody, which is complicated and random cross-linking process. As results, it causes decreases or loss of functional activity of either antibody or enzyme. In addition, most enzyme assays use secondary antibody to detect antigen binding activity of primary antibody. Enzymes coupled to secondary antibody provide a binding signal by substrate-based color development, suggesting secondary antibody is required in enzyme immunoassay. Additional incubation time for binding of secondary antibody should also be necessary. More importantly, non-specific binding activity caused by secondary antibody should also be eliminated. In this study, we cloned AP isolated from Escherichia coli (E. coli) chromosome by PCR and fused to) hAY4 single-chain variable domain fragment (ScFv) specific to death receptor (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand (TRAIL). hAY4 ScFv-AP expressed in E. coli showed 73.8 kDa as a monomer in SDS-PAGE. However, this fusion protein shown in size-exclusion chromatography (SEC) exhibited 147.6 kDa as a dimer confirming that natural dimerization of AP by non-covalent association induced ScFv-AP dimerization. In several immunoassay such as ELISA, Western blot and immunocytochemistry, it showed antigen binding activity by color development of substrates catalyzed by AP directly fused to primary hAY4 ScFv without secondary antibody. In summary, hAY4 ScFv-AP fusion protein was successfully purified as a soluble dimeric form in E. coli and showed antigen binding activity in several immunoassays without addition of secondary antibody which sometimes causes time-consuming, expensive and non-specific false binding.

A study on characteristics of palace wallpaper in the Joseon Dynasty - Focusing on Gyeongbokgung Palace, Changdeokgung Palace and Chilgung Palace - (조선시대 궁궐 도배지 특성 연구 - 경복궁, 창덕궁, 칠궁을 중심으로 -)

  • KIM Jiwon;KIM Jisun;KIM, Myoungnam;JEONG Seonhwa
    • Korean Journal of Heritage: History & Science
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    • v.56 no.1
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    • pp.80-97
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    • 2023
  • By taking wallpaper specimens from Gyeongbokgung Palace, Changdeokgung Palace, and Chilgung Palace preserved from the late Joseon Dynasty to the present, we planned in this study to determine the types and characteristics of the paper used as wallpaper in the Joseon royal family. First, we confirmed the features of paper hanging in the palaces with old literature on the wallpaper used by the royal family based on archival research. Second, we conducted a field survey targeting the royal palaces whose construction period was relatively clear, and analyzed the first layer of wallpaper directly attached to the wall structure after sampling the specimens. Therefore, we confirmed that the main raw material was hanji, which was used as a wallpaper by the royal family, and grasped the types of substances(dyes and pigments) used to produce a blue color in spaces that must have formality by analyzing the blue-colored paper. Based on the results confirmed through the analysis, we checked documents and the existing wallpaper by comparing the old literature related to wallpaper records of the Joseon Dynasty palaces. We also built a database for the restoration of cultural properties when conserving the wallpaper in the royal palaces. We examined the changes in wallpaper types by century and the content according to the place of use by extracting wallpaper-related contents recorded in 36 cases of Uigwe from the 17th to 20th centuries. As a result, it was found that the names used for document paper and wallpaper were not different, thus document paper and wallpaper were used without distinction during the Joseon Dynasty. And though there are differences in the types of wallpaper depending on the period, it was confirmed that the foundation of wallpaper continued until the late Joseon Dynasty, with Baekji(white hanji), Hubaekji(thick white paper), jeojuji(common hanji used to write documents), chojuji(hanji used as a draft for writing documents) and Gakjang(a wide and thick hanji used as a pad). As a result of fiber identification by the morphological characteristics of fibers and the normal color reaction(KS M ISO 9184-4: Graph "C" staining test) for the first layer of paper directly attached to the palace wall, the main materials of hanji used by the royal family were confirmed and the raw materials used to make hanii in buildings of palaces based on the construction period were determined. Also, as a result of analyzing the coloring materials of the blue decorative paper with an optical microscope, ultraviolet-visible spectroscopic analysis(UV-Vis), and X-ray diffraction analysis(XRD), we determined that the type of blue decorative paper dyes and pigments used in the palaces must have formality and identified that the raw materials used to produce the blue color were natural indigo, lazurite and cobalt blue.

Screening and Possibility of Semi-quantitative Analysis of Explosive Compounds in Soil Using EXPRAY$^{(R)}$ Explosives Field Detection Kit (화약물질 현장검출시약 EXPRAY$^{(R)}$를 이용한 토양내 화약물질 스크리닝 및 준정량화 가능성)

  • Bae, Bum-Han;Cho, Jung-Hyun
    • Journal of Soil and Groundwater Environment
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    • v.14 no.2
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    • pp.45-53
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    • 2009
  • A quick and simple detection method of explosive compounds in environmental matrix (soil and water) can provide a screening step which reduces the number of unnecessary samples and the cost of expensive laboratory analysis at a site investigation. A commercially available EXPRAY$^{(R)}$Explosives Field Detection Kit (EXPRAY) was used to determine the minimum detection concentration and to test the possibility of semi-quantitative analysis of 14 explosive compounds using standard solutions. The results showed that EXPRAY could detect 5 explosive compounds, TNT, RDX, HMX, Tetryl, and TNB, out of 14 US EPA designated explosives. The minimum detection limit of the nitramine explosives was 14 ng/$^2$ for HMX and RDX. EXPRAY was more sensitive to nitroaromatics than the nitramines and the minimum detection limits per unit area (mm$^2$) for Tetryl, TNB, and TNT, were 3 ng, 3 ng, and 0.3 ng, respectively. The semi-quantification of 5 explosive compounds in an order ofmagnitude could be achieved by the intensity of developed color only when EXPRAY was applied on the standard solutions under controlled laboratory conditions. With contaminated soil samples, however, only the presence and type of explosive compounds was identified. Therefore, EXPRAY is an economic and sensitive method that can be used in a screening step for the identification of explosives in the field samples.

Standardization for Analysis Method of Total Polyphenol in Complex of Picao Preto (피카오프레토 등 복합물 중 총 폴리페놀 분석법 표준화)

  • Hu, Soojung;Kim, Ji-An;Moon, Myung-Hee;Lee, Sung-Hye;Yoon, Hae-Seong;Hong, Jin-Hwan
    • Journal of Food Hygiene and Safety
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    • v.33 no.1
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    • pp.44-49
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    • 2018
  • As generic health functional food items have been expanded, this research project has been conducted to prepare a scientific and systematic standardized analytical method of relevant food item and examine the suitability of the method for health/functional foods on sale. Total polyphenol was necessary for development and verification of standardized analytical method. The method exhibited high linearity in the tannic acid calibration curve ($r^2$ > 0.999) over concentrations of $5-50{\mu}g/mL$. The limits of detection and quantitation for tannic acid were $5{\mu}g/mL$ and $15{\mu}g/mL$, respectively, while tannic acid recovery was 102.3-112.4% with standard deviations of 0.8-3.2%. To verify the accuracy of the analytical method, the labeled amounts of purchased health functional foods were monitored. The recovery for tannic acid was 105.6% of the labeled amounts. Thus, the new method was suitable for all cases.

Gemological Characterization of B. C. Jade (비씨 제이드의 보석학적 연구)

  • Kim, Won-Sa;Wight, Willow
    • Journal of the Mineralogical Society of Korea
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    • v.21 no.2
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    • pp.177-182
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    • 2008
  • The Gemological characteristics of B.C. jade from Cassiar Mine, British Colombia, Canada, have been investigated, using polarizing microscopy, Mohs' hardness, refractive index and density measurements, X-ray powder diffraction, X-ray fluorescence spectrometry, ICP-MS, Infrared absorption spectrometry, and DTA/TGA. The B.C. jade is deeply green (spinach peen or olive green) in color and is translucent. It shows a resinous or waxy luster. The principal mineral of the material is tremolite-actinolite solid solution and minor amount of Cr-garnet and unidentified opaque minerals are accompanied. Mohs' hardness value ($5.5{\sim}6$). refractive index (1.62), and specific gravity (3.01) are measured. It is very highly tough and shows hackly fracture. The high Fe content ($Fe_2O_3\;4.14{\sim}4.66\;wt%$) in B.C. jade is attributable to a deepening of green color of the material. The B.C. jade starts to dehydrate at v and dehydration is completed at $1000.8^{\circ}C$, transforming tremolite-actinolite solid solution to enstatite, diopside, quartz, and water in its place. This possible reaction is supported by the weight loss of B.C. jade (1.93 wt%) at $1000.8^{\circ}C$ indicated by TGA curve.

Analysis of gibberellic acid from fruits using HPLC/UV-vis (HPLC/UV-vis을 이용한 과일류 중의 지베렐린 산 분석)

  • Ma, Kyung Na;Cho, Hyun-Woo;Myung, Seung-Woon
    • Analytical Science and Technology
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    • v.26 no.1
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    • pp.19-26
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    • 2013
  • Gibberllic acid ($GA_3$) is one of gibberellins (GAs) that are a class of plant growth hormones that exert profound and diverse effects on plant growth and development. $GA_3$ is essentially non-UV absorbing and is difficult to assay by UV-detector. For effective extraction of gibberellic acid from fruits by using liquid-liquid extraction, optimized pH and extraction solvent were established. The selective and sensitive derivative of $GA_3$ for HPLC/UV-vis was derivatized using phenacyl bromide, and the experimental factors, including reaction time, reaction temperature and amount of derivatizing reagent and base were investigated for the effective synthesis. The derivatized $GA_3$ with phenacyl bromide was effectively analyzed by HPLC/UV-vis. The structure of derivatized $GA_3$ was confirmed by HPLC/ESI-MS. For apple, LOD and LOQ were 0.008 mg/kg and 0.027 mg/kg, respectively. For pear, LOD and LOQ were 0.003 mg/kg, 0.012 mg/kg, respectively. The established method can be applied to more effective analysis of $GA_3$ from plant and food.

Transformation of Populus alba $\times$Populus glandulosa Using Phosphinothricin Acetyltransferase Gene (Phosphinothricin acetyltransferase 유전자를 이용한 현사시의 형질전환)

  • 오경은;양덕춘;문흥규;박재인
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.163-169
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    • 1999
  • This study was conducted to produce herbicide resistant plants by transferring phosphinothricin acetyltransferase (PAT) gene into Populus alba $\times$ Populus glandulosa No .3 using Agrobacterium tumefaciens MP 90/PAT. Leaf segments from in vitro grown shoots of hybrid poplar No. 3 were soaked in a AB medium containing Agrobacterium tumefaciens MP 90/PAT for 10 min and cocultivated for 2 days on MS medium containing 1.0 mg/L 2,4-D and 0.2mg/L kinetin (CIM). Putative transformed calli could be selected after cocultivation of leaf segments on CIM supplemented with 50mg/L kanamycin and 500mg/L cefotaxime for 3 weeks. The selected calli were cultured on CIM supplemented with 50 mg/L kanamycin and 500 mg/L cefotaxime for 5~8 weeks before transfer to WPM containing 1.0mg/L zeatin, 0.1mg/L BAP, 50 mg/L kanamycin and 500mg/L cefotaxime for shoot regeneration. Shoots were regenerated from the callus after 4 week cultivation, and the regenerants were grown on the same medium for 7~l0 weeks. The plants rooted on 1/2 WPM containing 0.2 mg/L IBA and 50 mg/L kanamycin. To confirm the gene insertion into plants, GUS activity was detected by histochemical assay in the transformed plants. Finally, the presence of both NPT II and PAT genes from the transgenic plants were confirmed by PCR amplification with the gene specific primers and subsequent PCR-Southern blot with DIG-labeled PAT gene probe. After acclimatization in pots for 4 weeks, the plants were sprayed by 3 mL/L of Basta to test resistance to the herbicide. The transgenic plants remained green, whereas all the control plants died after one week.

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