• Title/Summary/Keyword: simultaneous recovery

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In situ Recovery of hGM-CSF in Transgenic Rice Cell Suspension Cultures (형질전환 벼 현탁세포 배양에서 hGM-CSF의 in situ Recovery 연구)

  • Myoung, Hyun-Jong;Choi, Hong-Yeol;Nam, Hyung-Jin;Kim, Dong-Il
    • KSBB Journal
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    • v.30 no.3
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    • pp.103-108
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    • 2015
  • Production of foreign proteins by transgenic plant cell cultures has several advantages such as post-translational modification, low risk of product contamination and low-cost production and purification. However, target proteins are degraded by extracellular proteases existing in the media. A solution to this problem is the use of perfusion culture and ion exchange chromatography for the application of integrated bioprocess using in situ recovery. With this method, production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was investigated in this study. First, optimization of cell concentration during the induction phase for the production of hGM-CSF was examined. As cell concentration increased, the level of hGM-CSF was decreased due to the presence of extracellular proteases. Induction using sugarfree media produced 33% more hGM-CSF. The effects of pH on the binding of hGM-CSF to cationic and anionic exchange resins were also investigated. In terms of stability, optimal pH was found to be 5~7. In the case of using buffer exchange when CM-Sepharose was used as a cationic exchange resin, optimal pH for binding was 4.8 and adsorption yield was 77%. When DEAE-Sepharose was used as an anionic exchange resin, it was 5.5 (74%). Without buffer exchange, optimal pH was 4.6 (84%). From these results, an integrated bioprocess using in situ recovery with simultaneous production and separation of foreign protein in transgenic plant cell suspension cultures was found to be feasible.

Simultaneous Unwrapping Phase and Error Recovery from Inhomogeneity (SUPER) for Quantitative Susceptibility Mapping of the Human Brain

  • Yang, Young-Joong;Yoon, Jong-Hyun;Baek, Hyun-Man;Ahn, Chang-Beom
    • Investigative Magnetic Resonance Imaging
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    • v.22 no.1
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    • pp.37-49
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    • 2018
  • Purpose: The effect of global inhomogeneity on quantitative susceptibility mapping (QSM) was investigated. A technique referred to as Simultaneous Unwrapping Phase with Error Recovery from inhomogeneity (SUPER) is suggested as a preprocessing to QSM to remove global field inhomogeneity-induced phase by polynomial fitting. Materials and Methods: The effect of global inhomogeneity on QSM was investigated by numerical simulations. Three types of global inhomogeneity were added to the tissue susceptibility phase, and the root mean square error (RMSE) in the susceptibility map was evaluated. In-vivo QSM imaging with volunteers was carried out for 3.0T and 7.0T MRI systems to demonstrate the efficacy of the proposed method. Results: The SUPER technique removed harmonic and non-harmonic global phases. Previously only the harmonic phase was removed by the background phase removal method. The global phase contained a non-harmonic phase due to various experimental and physiological causes, which degraded a susceptibility map. The RMSE in the susceptibility map increased under the influence of global inhomogeneity; while the error was consistent, irrespective of the global inhomogeneity, if the inhomogeneity was corrected by the SUPER technique. In-vivo QSM imaging with volunteers at 3.0T and 7.0T MRI systems showed better definition in small vascular structures and reduced fluctuation and non-uniformity in the frontal lobes, where field inhomogeneity was more severe. Conclusion: Correcting global inhomogeneity using the SUPER technique is an effective way to obtain an accurate susceptibility map on QSM method. Since the susceptibility variations are small quantities in the brain tissue, correction of the inhomogeneity is an essential element for obtaining an accurate QSM.

Simultaneous Determination of Organo-phosphorus and Organo-chlorinated Pesticides in Soil and Lawn Samples of Golf Courses (골프장 토양 및 잔디 시료 중 유기인 및 유기염소계 농약의 동시 분석)

  • Seo, Yong-Chan
    • Journal of Korean Society of Environmental Engineers
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    • v.27 no.12
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    • pp.1257-1262
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    • 2005
  • Organo-chlorinated pesticides have been seldom measured in Korea because their use was banned for more than two decades. Nevertheless, the environmental persistency of these pesticides requires regular monitoring. Only organo-phosphorus pesticides are currently being monitored in Korea. Simultaneous determination of organo-chlorinated and organo-phosphorus pesticides was attempted using gas chromatography/mass spectrometer detector. Pressurized fluid extraction(PFE) was used in sample preparation. PFE showed equivalent extraction efficiency to traditional soxhlet extraction with less solvent use and less sample preparation time. Average recovery of spiked pesticides to soil and lawn samples was 62% ranging 52 to 154%. Method detection levels(MDLs) for these pesticides ranged from 3.4 to 10.5 ${\mu}g/kg$. Endosulfan, fenitrothion, and diazinon were found in the samples from 5 different golf courses out of 9 courses.

A Study on the Simultaneous Determination of Residual Zeranol, Zearalenone and Their Metabolites in Beef by Gas Chromatography/Mass Spectrometry (Gas Chromatography/Mass Spectrometry에 의한 우육 중의 잔류 Zeranol, Zearalenone 및 그 대사산물들의 동시 분석법에 대한 연구)

  • 이은섭;이용욱
    • Journal of Food Hygiene and Safety
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    • v.9 no.1
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    • pp.1-13
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    • 1994
  • A Simultaneous determination method was improved for the determination and confirmation of zeranol, zearalenone, as well as their isomers and metabolites, in beef. The analytes were extracted from tissue by CH3CN, hydrolyzed enzymatically(for glucuronide conjugates), cleaned up by a strong basic anion exchange resin combined with a liquid/liquid partitioning, derivatized using MSTFA and confirmed, quantified by GC/MS/SIM with a internal standard, zearalane. The results were as follows : (1) all the estrogens were separated on the GC/MS chromatogram under the extraction method and the chromatographic conditions improved, the retention times of zearalane-TMS2, zearalanone-TMS2, zearalenone-TMS2, zeranol-TMS3, taleranol-TMS3, and $\alpha$-zearalenol-TMS3, $\beta$-zearalenol-TMS3, were 18.49, 19.44, 19.63, 19.71, 19.79 and 19.99, 20.08 minutes, respectively. (2) The calibration curves of residual zeranol, zearalenone and their metabolites showed constantly linear(r=0.99) in the range of 5~20 ng. The minimum detection concentration of residual zeranol, zearalenone and their metabolites was 1 ppb. (3) The total average recovery of residual zeranol, zearalenone and their metabolites from spiked beef was 60.2%(CV=29.7%) at the 1 ppb and 63.5%(CV=26.5) at the 2 ppb, 72.9%(CV=18.2%) at the 4 ppb. (4) The preservation method for 6 estrogens was improved for the fast running time(21 min) and MSTFA was utilized for derivatizing 6 estrogens for improvement of recovery, for good resolution, for characteristic mass spectra unlike Jose's method and Tina's method. The utilization of zearalane as internal standard showed good quantification result for zeranol, zearalenone, as well as their isomers and metabolites, in beef.

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The Performance Degradation of PEMFCs Fabricated with Different GDLs During Exposure to Simultaneous Sulfur Impurity Poisoning Condition (서로 다른 GDL을 이용한 고분자전해질 막 연료전지의 황불순물 복합피독에 의한 성능 저하)

  • Lee, Soo;Kim, Jae-Hyun;Jin, Seok-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.30 no.1
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    • pp.146-151
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    • 2013
  • This paper reveals the performance decrease and recovery of PEMFC when the contaminated fuel gas and air source with sulfur impurities such as hydrogen sulfide and sulfur dioxide were simultaneously introduced to anode and cathode, respectively. Three different GDLs were fabricated with different carbon black and activated carbon to prevent an introduction of sulfur compound impurities into MEA. components. The severity of $SO_2$ and $H_2S$ poisoning was depended on concentrations(3 ppm - 10 ppm) of sulfur impurities. Especially, cell performance degradation rate was rapid when MEA fabricated with CN-2 GDL because it had little porosity on GDL surface. Moreover, the cell performance can be recovered up to 90%-95% only with neat hydrogen and fresh air feeding.. Conclusively, MEA fabricated with porous CN-1 GDL showed the best cell performance and recovery efficiency during exposure to poisoning condition by simultaneous sulfur impurities.

A role of Sodium Bicarbonate Cotransporter(NBC) in $HCO_3^-$ Formation in Human Salivary Gland Acinar Cells

  • Jin, Mee-Hyun;Koo, Na-Youn;Jin, Mei-Hong;Hwang, Sung-Min;Park, Kyung-Pyo
    • International Journal of Oral Biology
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    • v.33 no.4
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    • pp.143-147
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    • 2008
  • The sodium bicarbonate cotransporter (NBC) protein is functionally expressed in salivary glands. In this experiment, we examined the role of NBC in $HCO_3^-$ formation in human parotid gland acinar cells. Intracellular pH (pHi) was measured in 2'-7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF)-loaded cells. Acetazolamide (0.1 mM) and 4,4'-diisothio cyanatostilbene-2,2'-disulphonic acid (DIDS, 0.5 mM) were used as specific inhibitors of carbonic anhydrase and NBC, respectively. The degree of inhibition was assessed by measuring the pHi recovery rate (${\Delta}pHi$/min) after cell acidification using an ammonium prepulse technique. In control experiments, ${\Delta}pHi$/min was $1.40{\pm}0.06$. Treatment of cells with 0.5 mM DIDS or 0.1 mM acetazolamide significantly reduced ${\Delta}pHi$/min to $1.14{\pm}0.14$ and $0.74{\pm}0.15$, respectively. Simultaneous application of DIDS and acetazolamide further reduced ${\Delta}pHi$/min to $0.47{\pm}0.10$. Therefore, DIDS and acetazolamide reduced ${\Delta}pHi$/min by 19% and 47%, respectively, while simultaneous application of both DIDS and acetazolamide caused a reduction in ${\Delta}pHi$/min of 67%. These results suggest that in addition to carbonic anhydrase, NBC also partially contributes to $HCO_3^-$ formation in human parotid gland acinar cells.

New Method for Simultaneous Quantification of 12 Ginsenosides in Red Ginseng Powder and Extract: In-house Method Validation

  • In, Gyo;Ahn, Nam-Geun;Bae, Bong-Seok;Han, Sung-Tai;Noh, Kil-Bong;Kim, Cheon-Suk
    • Journal of Ginseng Research
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    • v.36 no.2
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    • pp.205-210
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    • 2012
  • For quality control of components in Korean red ginseng powder and extract, a new method for simultaneous quantification of 12 ginsenosides ($Rg_1$, Re, Rf, $Rh_1$, $Rg_2$[S], $Rg_2$[R], $Rb_1$, Rc, $Rb_2$, Rd, $Rg_3$[S], and $Rg_3$[R]) was studied. Compared to the official method for quantification of marker substances (ginsenosides $Rg_1$ and $Rb_1$), the proposed methods were guaranteed by in-house method validation. Several criteria such as linearity, specificity, precision and accuracy were evaluated. For red ginseng powder, recovery (averaging 95% to 105%) was calculated, and analysis of variance was carried out to estimate the relative standard deviation (0.20% to 2.12%). For red ginseng extract, the average recovery rate was 90% to 99% and the relative standard deviation was 0.39% to 2.40%. These results indicate that the proposed method could be used in the laboratory for determination of 12 ginsenosides in red ginseng powder and extract. In addition, this method was found to be suitable for quality control of ginseng products and potentially offer time and cost benefits.

Determination of Coccidiostats (Amprolium and Decoquinate) in Cattle and Chicken's Muscle using High Performance Liquid Chromatography

  • Kim, Byung-Ju;Ham, Hyun-Sun;Lee, Jin-Joo;Cheong, Nam-Yong;Myung, Seung-Woon
    • Bulletin of the Korean Chemical Society
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    • v.33 no.2
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    • pp.559-563
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    • 2012
  • An analytical method for the simultaneous determination of veterinary medicines (amprolium and decoquinate) in cattle and chicken's muscle by HPLC/UV-vis was established. Samples were extracted by a HLB (Hydrophilic-Liphophilic Balance) cartridge with acetonitrile and methanol. Prior to HPLC injection, a mixture solvent (Water:MeOH, 1:1) was utilized as a reconstitution solvent. Chromatographic separation was achieved with a C18 column ($250{\times}4.6mm$, $5{\mu}m$) using gradient elution with 20 mM HFBA and MeOH:ACN (1:1.8). The calibration curves from the spiked blank matrix showed good linearity (above $r^2$=0.997) in the concentration range of $0.13-12.0mg\;kg^{-1}$. The relative recovery (accuracy) and limit of quantitation (LOQ) were in the range of 78.5-107.1% and $0.13-0.42mg\;kg^{-1}$, respectively. The developed method can be used to determine under the MRL (Maximum Residue Limits) levels of veterinary medicines in animal tissues.

Electrodeposition of Graphene-Zn/Al Layered Double Hydroxide (LDH) Composite for Selective Determination of Hydroquinone

  • Kwon, Yeonji;Hong, Hun-Gi
    • Bulletin of the Korean Chemical Society
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    • v.34 no.6
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    • pp.1755-1762
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    • 2013
  • A graphene-Zn/Al layered double hydroxide composite film was simultaneously prepared by electrochemical deposition on the surface of a glassy carbon electrode (G-LDH/GCE) from the mixture solution containing GO and nitrate salts of $Zn^{2+}$ and $Al^{3+}$. The modified electrode showed good electrochemical performances toward the simultaneous electrochemical detection of hydroquinone (HQ), catechol (CA) and resorcinol (RE) due to the unique properties of graphene (G) and LDH such as large active surface area, facile electronic transport and high electrocatalytic activity. The redox characteristics of G-LDH/GCE were investigated with cyclic voltammetry and differential pulse voltammetry. The well-separated oxidation peak potentials, corresponding to the oxidation of HQ, CA and RE, were observed at 0.126 V, 0.228 V and 0.620 V respectively. The amperometric response of the modified electrode exhibited that HQ can be detected without interference of CA and RE. Under the optimized conditions, the oxidation peak current of HQ is linear with the concentration of HQ from 6.0 ${\mu}M$ to 325.0 ${\mu}M$ with the detection limit of 0.077 ${\mu}M$ (S/N=3). The modified electrode was successfully applied to the direct determination of HQ in a local tap water, showing reliable recovery data.

The Effects of Glycerol Aftertreatment for Low-Formaldehyde Finishing (Glycerol 후처리에 의한 Free-formaldehyde 발생 억제 효과)

  • Choi Suk-Chul;Kim Ho-Jung
    • Journal of the Korean Society of Clothing and Textiles
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    • v.10 no.2
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    • pp.59-67
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    • 1986
  • To control the amounts of formaldehyde released from the cotton fabric finished with N-methylol compounds, glycerol was used as a formaldehyde-scavenging agent. It was observed the effects of catalysts and curing conditions when aftertreated with glycerol on melamine resin finished fabric. The effects of Different processes of glycerol treatment, and different resins, urea resin and melamine resin, were compared. The conclusions obtained from the results are as follows: 1) It was shown hatt the aftertreatment with glycerol (treated without catalyst) was more effective than treated with catalysts in controlling free formaldehyde. 2) The optimum curing temperature and curing time for the glycerol aftertreatment without adversely affecting the other properties of fabric was about $160^{\circ}C$, 3 min. 3) According to the increase of glycerol concentration in both aftertreatment and simultaneous treatment the amounts of free formaldehyde was reduced. The rate of decrease was manifest within the limits of $6\%$ in the case of simultaneous treatment with glycerol ana resins, and $3\%$ in the case of glycerol aftertreatment on resin finished fabrics. 4) Dry wrinkle recovery angle was decreased the increase of glycerol concentration. Melamine resin had a little adverse effect than urea resin, particulary glycerol aftertreatment. 5) The breaking strength was increased with the increase of glycerol concentration.

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