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Comparative N-Linked Glycan Analysis of Wild-Type and α1,3-Galactosyltransferase Gene Knock-Out Pig Fibroblasts Using Mass Spectrometry Approaches

  • Park, Hae-Min;Kim, Yoon-Woo;Kim, Kyoung-Jin;Kim, Young June;Yang, Yung-Hun;Jin, Jang Mi;Kim, Young Hwan;Kim, Byung-Gee;Shim, Hosup;Kim, Yun-Gon
    • Molecules and Cells
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    • v.38 no.1
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    • pp.65-74
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    • 2015
  • Carbohydrate antigens expressed on pig cells are considered to be major barriers in pig-to-human xenotransplantation. Even after ${\alpha}1,3$-galactosyltransferase gene knock-out (GalT-KO) pigs are generated, potential non-Gal antigens are still existed. However, to the best of our knowledge there is no extensive study analyzing N-glycans expressed on the GalT-KO pig tissues or cells. Here, we identified and quantified totally 47 N-glycans from wild-type (WT) and GalT-KO pig fibroblasts using mass spectrometry. First, our results confirmed the absence of galactose-alpha-1,3-galactose (${\alpha}$-Gal) residue in the GalT-KO pig cells. Interestingly, we showed that the level of overall fucosylated N-glycans from GalT-KO pig fibroblasts is much higher than from WT pig fibroblasts. Moreover, the relative quantity of the N-glycolylneuraminic acid (NeuGc) antigen is slightly higher in the GalT-KO pigs. Thus, this study will contribute to a better understanding of cellular glycan alterations on GalT-KO pigs for successful xenotransplantation.

Mixture Toxicity Test of Ten Major Chemicals Using Daphnia magna by Response Curve Method (독성 반응곡선을 이용한 수계 주요 오염물질의 혼합독성평가)

  • Ra, Jin-Sung;Kim, Ki-Tae;Kim, Sang-Don;Han, Sang-Guk;Chang, Nam-Ik;Kim, Yong-Seok
    • Journal of Korean Society of Environmental Engineers
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    • v.27 no.1
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    • pp.67-74
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    • 2005
  • Toxicity tests were performed to evaluate the feasibility of application with prediction models to 10 mixture chemicals (chloroneb, butylbenzylphthalate, pendimethaline, di-n-butylphthalate, di-iso-butylphthalate, diazinon, isofenphos, 2-chlorophenol, 2,4,6-trichlorophenol and p-octylphenol) detected in effluents from wastewater treatment plants (WWTPs). Ten chemicals were selected in the basis of their toxicities to Daphnia magna and the concentrations in effluents measured by GC/MS. Three models including concentration addition (CA), independent action (IA) and effect summation (ES) were employed for the comparison of the predicted and the observed mortality of D. magna exposed to 10 mixture chemicals for 48 hours. With a comparative study it was ineffective to predict the mortality through the CA and the ES prediction model, while the IA prediction model showed a high correlation($r^2\;=\;0.85$). Moreover, the ES model over-estimated the toxicity observed by bioassay experiments about five-fold. Consequently, IA model is a reasonable tool to predict the mixture toxicity of the discharging water from WWTPs.

A Study on the Characteristic Trace of Water Quality Pollutants in the Industrial Wastewater (업종별 산업폐수의 수질오염물질 배출 특성)

  • Park, Sun Ku;Kim, Sung Soo;Ko, Oh Suk
    • Analytical Science and Technology
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    • v.12 no.2
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    • pp.141-150
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    • 1999
  • Twenty organic chemical substances, tetrachloroethylene, toluene, ethylbenzene, p-xylene, m-xylene, isopropyl benzene, stylene, bromobenzene, 1,3,5-trimethylbenzene, 2-chlorotoluene 1,2,4-trimethylbenzene, p-isopropyltoluene, 4-chlorotoluene, n-butylbenzene, 1,2,4-trichlorobenzene, naphthalene, tert-butylbenzene, sec-butylbenzene, phenol, isopropyl benzene hydroperoxide were isolated from untreated and treated wastewater collected at 76 companys of 9 types industry in the basin of Young San River. Their organic compounds were elucidated by Gas Chromatography/Mass Spectrometry (GC/MS) and by comparison with each standard reagents. Especially, phenol compound is detected from effluent water but not detected from plant wastewater in the chemical industry. Heavy metal, which are Cr, Mn, Cu, Zn, Cd, Pb, As, Al and Fe, are contained in the plant wastewater of all industry, Fe, Al of them is more detected than the other metals in plant wastewater. Cr, Cd, Pb, As is contained much in plant wastewater of electricity and electron, metal molding industry. Nine metals is nearely treated when plant wastewater is treated, and then the concentration of each other metals is detected below water quality standard or not detected by using AA.

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Synthesis and Isolation of Monoacetyl-DCB and Diacetyl-DCB from 3,3대-dichlorobenzidine(DCB) (디클로로벤지딘으로부터 대사물질의 합성과 분리방법에 대한 연구)

  • Lee, Jin-Heon;Lee, Beom-Gyu
    • Journal of Environmental Health Sciences
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    • v.29 no.2
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    • pp.50-55
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    • 2003
  • 3,3-dichlorobenzidine is suspected to be cancinogenic in experimental animal and human. Several studies have investigated excretion of metabolites in urine, hemoglobin adduction and cancer incidence among workers occupationally exposed to 3,3'-dichlorobenzidine. In these researches, metabolites of 3,3'-dichlorobenzidine had a very important role, and were required as highly purity. The purpose of this study was synthesis and isolation of its metabolites from 3,3'-dichlorobenzidine. 3,3'-dichlorobenzidine was partially dissolved in benzene, ether, ethanol and methanol, and completely dissolved in 70% acetic acid on mixtures of citric acid containing less than 1% DCB, pyridine, a mixture of 0.5N NaOH and toluene(1:2), and phenol saturated with 20 mM TRIZA base. DCB, monoacetyl-DCB and diacetyl-DCB were measured by using gas chromatography/mass spectrometry(GC/MS). Detection for checking them was nitrogen phosphorous detection mode(NPD), and for identifying them was selected ion monitoring mode(SIM). The base peaks were 252 m/z in DCB, 252, and 294 m/z in monoacetyl-DCB, and 252, 294 and 336 m/z in diacetyl-DCB, respectively. Diacetyl-DCB was synthesized by titrating DCB solution of pyridine with sufficient acetyl chloride. Precipitation was diacetyl-DCB, which was purity of 98.7%. And its supernatant was composed of DCB, monoacetyl-DCB and diacetyl-DCB. By using acetic acid as controller of acetylation, monoacetyl-DCB was isolated from diacetyl-DCB . And residual pyridine was removed by using acetone. The purity of monoacetyl-DCB was 98.8%.

Development of Analytical Method and Monitoring for Bifenazate in Commercial Agricultural Products (농산물 중 Bifenazate의 분석법 개선 및 모니터링)

  • Park, Eun-Heui;Go, Myoung-Jin;Cho, Myong-Shik;Kim, Young-Sun;Lee, Jin-Ha;Choi, Dong-Mi
    • The Korean Journal of Pesticide Science
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    • v.14 no.1
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    • pp.21-29
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    • 2010
  • The analytical method for bifenazate was developed using a HPLC (UVD). Also, analytical condition of LC/MS was set up for bifenazate. We validated the method for the precision and the reproducibility. The correlation coefficient of bifenazate ranged from 0.05 to 2.5 mg/kg was 1.0. Limit of quantitation (LOQ) was 0.01 mg/kg. To measure recoveries from agricultural products such as foxtail millet (cereal grains), kidney bean (beans), orange (fruits), perilla leaves (vegetables) and oak mushroom (mushrooms), bifenazate was spiked. Mean recoveries of bifenazate for each sample were 82.7~104.1% at the level of 0.1 mg/kg and 73.1~104.3% at the level of 0.5 mg/kg. The relative standard deviations (n=3) were 0.2~9.7%. Pesticide residues for bifenazate were investigated in 16 commodities (rice, foxtail millet, buckwheat, kidney bean, peanut, sesame, orange, grapefruit, kiwifruit, spinach, perilla leaves, leek, garlic stem, garlic, ginger and oak mushroom) collected from 22 provinces in 2009. Bifenazate was analyzed using analytical method by HPLC from 304 samples, and residue was not detected.

Isolation and Identification of Antimicrobial Compound from Amarantus lividus (참비름 추출물에서 항균성 물질의 분리 및 동정)

  • Oh, Young-Sook;Lee, Shin-Ho
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.123-129
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    • 2005
  • Isolation and identification of pathogens from slaughter and meat processing plant were investigated. Antimicrobial activity of Amaranthus lividus against isolated pathogens such as Aeromonas sobria, Escherichia coli, Escherichia coli O157, Listeria monocytogenes, Salmonella spp., and Staphylococcus aureus was investigated. Among the chloroform, ethyl acetate and buthanol fraction of amaranthus lividus showed inhibitory effect against Aeromonas sobria CLFM1 and Escherichia coli CLFM2. Antimicrobial substance in chloroform fraction was isolated by silica gel adsorption column chromatography, sephadex LH-20 column chromatography and silica gel partition column chromatography. The antimicrobial compound of amaranthus lividus was identified as diethyl phtalate by HPLC, GC-MS, H-NMR and C-NMR.

Volatile Flavor Components in Various Edible Portions of Angelica keiskei Koidz (신선초의 식용부위별 향기성분)

  • Park, Eun-Ryong;Lee, Hae-Jung;Lee, Myung-Yul;Kim, Kyong-Su
    • Korean Journal of Food Science and Technology
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    • v.29 no.4
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    • pp.641-647
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    • 1997
  • Volatile flavor components in whole edible portion, stem and leaf of fresh angelica (Angelica keiskei Koidz) were extracted by SDE (simultaneous steam distillation and extraction) method using the mixture of n-pentane and diethylether (1:1, v/v) as an extract solvent and analyzed by GC-FID and GC/MS. Identification of the volatile flavor components in aroma concentrate was mostly based on the RI of GC and mass spectrum of GC/MS. Twenty five hydrocarbons, 15 alcohols, 3 aldehydes, 6 esters, 2 ketones and 1 acid were identified in the whole edible portion of angelica. Twenty hydrocarbons, 13 alcohols, 4 esters and 1 acid were identified in the stem sample of angelica. Nineteen hydrocarbons, 11 alcohols, 4 aldehydes, 6 esters, 2 ketones and 1 acid were identified in the leaf sample of angelica. ${\gamma}-Terpinene$, germacrene B, ${\delta}-3-carene$, cis-3-hexen-1-ol, ${\gamma}-muurolene$ and ${\gamma}-elemene$ were the main components in each edible portions of angelica. The terpenoid compounds in volatile flavor components identified from whole edible portion, stem and leaf samples were confirmed as 75.76%, 86.42% and 78.21%, respectively. These results suggest that terpenoid compounds have a great effect on the flavor characteristics of angelica.

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Effect of Electron-Beam Irradiation on Flavor Components in Pear (Pyrus pyrifolia cv. Niitaka) (전자선 조사가 신고배의 향기성분에 미치는 영향)

  • Kim, Won;Shim, Sung-Lye;Ryu, Keun-Young;Jun, Sam-Nyeo;Jung, Chan-Hee;Seo, Hye-Young;Song, Hyun-Pa;Kim, Kyong-Su
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.2
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    • pp.195-202
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    • 2008
  • This study was performed to examine the effects of electron beam irradiation on volatile flavor components of Shingo pear (Pyrus pyrifolia cv. Niitaka) and on their changes according to storing period following irradiation. Volatile flavor components in pear were extracted using simultaneous steam distillation and extraction (SDE) apparatus and analyzed by GC/MS. 46 components were identified in control whereas 45, 44, 48 and 51 components were identified in irradiated samples by electron beam at 0.25, 0.5, 1, and 3 kGy, respectively. Hexanal, n-hexanol, and (E)-2-hexenal were identified as the major volatile flavor components of all samples. The characteristic volatile flavor components of irradiated pear by electron beam were similar to those of control, and their effects depending on irradiation source were not different. In addition, there was no noticeable change in volatile flavor components of pear with storage at $4^{\circ}C$ for 30 days or with irradiation. Sensory evaluation indicated that the consumer receptiveness tended to be higher at a low level of radiation dose under 1 kGy than control, albeit not significant. Therefore, electron beam irradiation at low level of radiation dose under 1 kGy could be considered as an effective method to exterminate vermin and thus to improve the shelf-stability of pear without deterioration.

ELECTROPHYSIOLOGICAL AND MORPHOLOGICAL CHARACTERISTICS OF FACIAL NUCLEUS IN RAT (흰쥐 안면신경핵 세포의 전기생리학적 및 형태학적 특성)

  • Choi, Byung-Ju;Cho, Jin-Hwa;Bae, Yong-Chul;Kim, Young-Jin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.27 no.3
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    • pp.400-409
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    • 2000
  • This study used in vivo intracellular and extracellular field potential recording to evaluate the intrinsic membrane properties and connection pattern within facial nucleus. 1. There were four subdivisions of medial, intermediate, lateral, and dorsolateral in facial nucleus. 2. Principal cells in the facial nucleus was recorded from and filled with neurobiotin in anesthetized rats. The extent of their dendrites and the characteristics of cell body were examined. 3. Principal cells had a large amplitude action potential and afterhyperpolarization was followed a single action potential. 4. The response from facial motonucleus to electrical stimulation of the facial nerve was mainly a monophasic wave, with a latency of 1 msec, which was assumed to reflect antidromic activation of facial motoneurons. In some of rats the response in addition showed late components at a latency of about 7-8 msec, but its amplitude was small. 5 Most of cells exhibited accommodation of spike discharge upon depolarization of membrane by 0.8 nA for 400 ms. Our results support the hypothesis that there normally are weak connections between different parts of the facial motonucleus to explain pathophysiology of hemifacial spasm and facial naive paralysis.

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Rancidity Analysis of Rapeseed Oil under Different Storage Conditions Using Mass Spectrometry-based Electronic Nose (질량분석기 기반-전자코를 이용한 저장중 유채유의 산패 분석)

  • Hong, Eun-Jeung;Lim, Chae-Lan;Son, Hee-Jin;Choi, Jin-Young;Noh, Bong-Soo
    • Korean Journal of Food Science and Technology
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    • v.42 no.6
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    • pp.699-704
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    • 2010
  • Rapeseed oil was stored under different conditions such as in the dark, with UV treatment, and with prooxidantscytochrome C and copper ion. The rapeseed oils stored at different temperatures were analyzed by a mass spectrometrybased electronic nose and discriminant function analysis (DFA). Volatile components in the rapeseed oil increased with storage time, and the discriminant function first score (DF1) moved from a positive position to a negative position as storage time increased. Changes in DF1 were higher under UV treatment than under the dark condition (DF1: $r^2$=0.9481, F=307.03). The different DF1 values (F1) under the dark condition were 0.099, 0.187, and 0.278 as storage temperature increased. The different values under UV treatment were 0.554, 0.588, and 0.542, as storage temperature increased from 4 to $26^{\circ}C$. As concentrations of prooxidants copper ion and cytochrome C increased, amounts of volatile components also increased. These were confirmed by DFA. Furthermore, changes in responses at each ion fragment agreed with reported results for GC/MS, which formed after rancidity of the oil, including pentane, pentanal, 1-pentanol, hexanal, n-octane, 2-hexenal, heptanal, 2-heptenal, decane, 2-octenal, undecane, and dodecane.