• Title/Summary/Keyword: GB 분리법

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Determination of plasma C16-C24 globotriaosylceramide (Gb3) isoforms by tandem mass spectrometry for diagnosis of Fabry disease (패브리병(Fabry) 진단을 위한 혈장 중 Globotriaosylceramide (Gb3)의 탠덤매스 분석법 개발과 임상 응용)

  • Yoon, Hye-Ran;Cho, Kyung-Hee;Yoo, Han-Wook;Choi, Jin-Ho;Lee, Dong-Hwan;Zhang, Kate;Keutzer, Joan
    • Journal of Genetic Medicine
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    • v.4 no.1
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    • pp.45-52
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    • 2007
  • Purpose : A simple, rapid, and highly sensitive analytical method for Gb3 in plasma was developed without labor-ex tensive pre-treatment by electrospray ionization MS/ MS (ESI-MS/MS). Measurement of globotriaosy lceramide (Gb3, ceramide trihex oside) in plasma has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme ${\alpha}$-galactosidase A (${\alpha}$-Gal A). The lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3. Methods : Only simple 50-fold dilution of plasma is necessary for the extraction and isolation of Gb3 in plasma. Gb3 in diluted plasma was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS. Results : Eight isoforms of Gb3 were completely resolved from plasma matrix. C16:0 Gb3 occupied 50% of total Gb3 as a major component in plasma. Linear relationship for Gb3 isoforms w as found in the range of 0.001-1.0 ${\mu}g$/mL. The limit of detection (S/N=3) was 0.001 ${\mu}g$/mL and limit of quantification was 0.01 ${\mu}g$/mL for C16:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9678 to 0.9982. Conclusion : This quantitative method developed could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.

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MT response on the two dimensional anisotropic structure (2차원 이방성 구조의 MT 반응)

  • Lee, Chun Gi;Gwon, Byeong Du
    • Journal of the Korean Geophysical Society
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    • v.2 no.2
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    • pp.123-134
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    • 1999
  • Magnetotelluric responses may be affected by strong anisotropy of the high-conductivity layers (HCL) in the upper mantle or lower crust. We have studied two-dimensional anisotropy MT modelling to examine the effect of high anisotropic media. Electrical properties of a homogeneous anisotropic body are defined by a symmetric conductivity tensor and the problem is described by coupled diffusion equation in the frequency domain. In two-dimensional anisotropic environments, diagonal elements of the impedance tensor have higher values than those in isotropic environments. In some cases, TM mode phases reach more than 90°and apparent resistivities decrease for some frequency range because of telluric distortion. GB decomposition may be used to recover regional responses, but can be affected by the regional anisotropic effect. Considering these results, BC87 dataset was interpreted with a modified anisotropic model.

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Relationship between Gb3 Expression and Cytotoxicity of Shiga-like Toxin I (Shiga-like Toxin I의 세포독성과 수용체 Gb3 발현과의 관계)

  • Lim, Suk-Hwan;Kim, Gi-Young;Kim, Hyung-Chun;Kim, Young-Hee;Son, Yong-Hae;Oh, Yang-Hyo;Park, Yeong-Min
    • Clinical and Experimental Pediatrics
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    • v.46 no.2
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    • pp.143-153
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    • 2003
  • Purpose : Infection with Shiga-like toxin (SLT)-producing Escherichia coli, an emerging human pathogen found particularly in young children under 5 years of age, causes a spectrum of illnesses with high morbidity and mortality, ranging from diarrhea to hemorrhagic colitis and hemolytic uremic syndrome. Host mediators play an important role in the pathogenesis of SLT-I toxicity. The experiments described here were designed to investigate the effect of SLT-I on TNF-${\alpha}$ production and to understand the effect of TNF-${\alpha}$ on GB3 expression. We also further examine the relationship between the Gb3 level and the differential susceptibility of cells to the cytotoxic action of SLT-I. Methods : The effect of purified SLT-1 from E. coli O157 : H7 (ATCC 43890) on tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) production in Raw264.7 cells was investigated. Many mediators regulate endothelial cell membrane expression of the glycolipid globotriaosyleramide (Gb3), which serves as the toxin receptor, suggesting that the host response to the toxin or other bacterial products may contribute to pathogenesis by regulating target cell sensitivity to the toxins. Therefore, the relationships between Gb3 expression and cytotoxicity against SLT-I on three types of cells were evaluated. Results : Detectable levels of TNF-${\alpha}$ were produced as early as six hours after induction and continued to increase during 48 hours by SLT-I. It was also found that Vero cells and dendritic cells (DC2.4 cells) expressed high levels of Gb3, 83% and 68%, respectively, and that Raw264.7 cells had a low level of Gb3 (29%) and appeared refractory to cytotoxicity against SLT-I. Vero cells and DC2.4 cells expressing high levels of Gb3 were highly susceptible to SLT-I. Furthermore, macrophages showed a resistance to SLT-I cytotoxicity, despite the fact that Gb3 expression was enhanced. Conclusion : These results strongly suggest that the expression of Gb3 is necessary but not sufficient to confer sensitivity of macrophages to SLT-I and further underpin the important role of SLT-I and its Gb3 receptors in the pathogenesis of E. coli O157 infection.

Regulation of Gb3 Expression on Dendritic Cells (수지상세포에 있어서 베로독소 수용체의 발현조절)

  • Lim, Suk-Hwan;Kim, Gi-Young;Kim, Hyung-Chun;Kim, Young-Hee;Son, Yong-Hae;Oh, Yang-Hyo;Park, Yeong-Min
    • Journal of Life Science
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    • v.17 no.4 s.84
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    • pp.482-492
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    • 2007
  • Infection with Shiga-like toxin (SLT)-producing Escherichia coli causes a spectrum of illnesses with high morbidity and mortality. Host mediators play an important role in the pathogenesis of SLT-I toxicity. We here investigated the effect of SLT-I on tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ production, effect of $TNF-{\alpha}$ on glycolipid globotriaosyleramide (Gb3) expression, and relationship between Gb3 level and differential susceptibility of cells to SLT-I. In this study, we observed that detectable levels of $TNF-{\alpha}$ are produced 6 hrs after induction and continued to increase during 48 hrs by SLT-I. It was also found that Vero cells and dendritic cells expressed high levels of Gb3, 83% and 68%, respectively, and that macrophages had a low level of Gb3 (29%) and showed refractory to cytotoxicity against SLT-I. Vero cells and dendritic cells expressing high levels of Gb3 were highly susceptible to SLT-I. furthermore, macrophages showed a resistance to SLT-I cytotoxicity, despite the fact that Gb3 expression was enhanced. These results suggest that the expression of Gb3 is necessary, but not sufficient to confer sensitivity of macrophages to SLT-I and further underpin the important role of SLT-I and its receptor, Gb3, in the pathogenesis of E. coli O157 infection.

Liquid-phase Microextraction Pretreatment Techniques for Analysis of Chemical Warfare Agents and Their Degradation Byproducts in Environmental Aqueous Samples (환경샘플 내 화학작용제 및 분해물질 분석을 위한 Liquid Phase Microextraction (LPME) 전처리 기법)

  • Kim, Dongwook;Chung, Wooyoung;Kye, Youngsik
    • Applied Chemistry for Engineering
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    • v.26 no.1
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    • pp.17-22
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    • 2015
  • International interests in chemical warfare agents (CWAs) have been increased recently because of the use of sarin (GB) in Syrian civil war which caused around 1,300 casualties in 2013. After exposing to natural environments, CWAs undergo hydrolysis or photodegrade to non-toxic degradation byproducts. Generally, CWAs and their degradation byproducts are present at very low concentration (e.g. several ppb), thus pretreatment processes including separation, extraction and concentration are required prior to any analyses. Liquid-liquid extraction and solid-phase extraction (SPE) are common techniques to pretreat environmental samples. Recently, a novel pretreatment method, liquid phase miecoextraction (LPME), has been applied to CWAs analysis, which could reduce amounts of solvent used but promote analytical efficiencies. Fundamental backgrounds of LPME and its application to CWAs analysis were reviewed.

Enzymatic preparation and antioxidant activities of protein hydrolysates from Gryllus bimaculatus (쌍별귀뚜라미 단백가수분해물의 제조 및 항산화 활성)

  • Cho, Hye-Rin;Lee, Yoo-Jung;Hong, Ji-Eun;Lee, Syng-Ook
    • Korean Journal of Food Science and Technology
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    • v.51 no.5
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    • pp.473-479
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    • 2019
  • Gryllus bimaculatus (GB) has recently been registered as a food variety in Korea. In the present study, we prepared protein hydrolysates from GB and evaluated their antioxidant capacity. Protein hydrolysates were prepared from dried GB using enzymatic hydrolysis using five different proteases, and protein hydrolysates showing high hydrolysis value (alcalase, flavourzyme, and neutrase) were separated further into fractions ${\leq}3kDa$ and then lyophilized. Based on $RC_{50}$ values of hydrolysates (${\leq}3kDa$) obtained from four different antioxidant analyses, the flavourzyme hydrolysates showed relatively high levels of antioxidant capacity among the three hydrolysates, and in particular, it showed considerably strong antioxidant activity in 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. The flavourzyme hydrolysate also significantly inhibited peroxidation of linoleic acid. These results suggest that protein hydrolysates from GB represent potential sources of natural antioxidants. Our current studies are focused on identification of active peptides from the flavourzyme hydrolysate.

Comparison of antioxidant capacity of protein hydrolysates from 4 different edible insects (식용곤충별 단백가수분해물의 항산화 활성 비교)

  • Jang, Hyun-Young;Park, Chae-Eun;Lee, Syng-Ook
    • Korean Journal of Food Science and Technology
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    • v.51 no.5
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    • pp.480-485
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    • 2019
  • The present study was conducted to compare antioxidant capacities of protein hydrolysates from four different edible insects (Protaetia brevitarsis larvae, Allomyrina dichotoma larvae, Gryllus bimaculatus imago, and Tenebrio molitor larvae) which have recently been registered as food varieties in Korea. Protein hydrolysates were prepared from each insect using enzymatic hydrolysis using alcalase, and were then separated into a fraction containing ${\leq}3kDa$. According to $RC_{50}$ values and trolox equivalent antioxidant capacity results obtained from five different antioxidant analyses, the Gryllus bimaculatus (GB) hydrolysate showed relatively high levels of antioxidant capacity and, in particular, the GB hydrolysate showed considerably strong antioxidant activities in 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and in ferric reducing antioxidant power (FRAP) assays. The GB hydrolysate also showed the strongest inhibitory effect on peroxidation of linoleic acid, and its rate of inhibition at $100{\mu}g/mL$ on day 3 of treatment was 60.26%. These results suggest that protein hydrolysates from edible insects including GB represent potential sources of natural antioxidants.

Serotype Distribution of Invasive Group B Streptococcal Diseases in Infants at Two University Hospitals in Korea (영아에서의 침습 B군 사슬알균 감염증 및 혈청형 분석: 2개 대학병원 연구)

  • Cho, Hye-Kyung;Nam, Hye Na;Cho, Hye Jung;Son, Dong Woo;Cho, Yong Kyun;Seo, Yiel-Hea;Kim, Yae-Jean;Eun, Byung Wook
    • Pediatric Infection and Vaccine
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    • v.24 no.2
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    • pp.79-86
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    • 2017
  • Purpose: This study was aimed at analyzing the serotypes of group B streptococcus (GBS) isolated from Korean infants with invasive disease and evaluating their association with disease manifestation. Methods: Data were retrospectively collected from invasive GBS infections at Gachon University Gil Medical Center from January 2006 to June 2012 and at Samsung Medical Center from April 2010 to November 2012. Serotypes were determined by slide agglutination test. Results: A total of 37 cases were identified, which included 22 full-term infants and 15 preterm infants. Fifteen cases (40.5%) were early-onset, 19 (51.4%) was late-onset, and three (8.1%) was very late-onset. Early-onset diseases among preterm infants were higher than those among full-term infants (60.0% [9/15] vs. 27.3% [6/22], P=0.17). The most common manifestation was bacteremia (70.3%), followed by meningitis and septic arthritis. Among 24 isolates retrievable for serotyping, serotype III (41.7%) was most common, followed by V (16.7%), Ia, Ib, and II (12.5%, respectively), and non-typeable (4.2%). Serotype III was more common in isolates from full-term infants (10/22) than from preterm infants (0/15), whereas serotype V was more common in isolates from preterm infants (4/15) than from full-term infants (0/22) (P=0.002). No penicillin-resistant strain was detected, and resistance to erythromycin and clindamycin were both 64.9%. Conclusions: GBS is an important pathogen in both preterm and full-term infants, and serotype distribution of GBS causing invasive diseases can differ between preterm and full-term infants. It is necessary to monitor the nationwide epidemiology of GBS diseases, including in preterm infants, in order to prepare preventive measures without underestimating early-onset diseases.