• Title/Summary/Keyword: Quadrupole Cell

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Application of Computer-coupled Mass Spectrometer for Continuous On-line Monitoring of Cell Growth and Growth Rate (세포증식과 증식속도의 On-line Monitoring을 위한 Computer- coupled Mass Spectrometer의 응용)

  • 남수완;최춘순;김정회
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.241-246
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    • 1989
  • Continuous on-line monitoring of cell concentration and growth rate in aerobic batch fermentation process was carried out by analyzing the exhaust gas composition of tormentor with a quadrupole mass spectrometer. From the mass spectrometric analyses of major gaseous components, i.e. $N_2$, $O_2$, $CO_2$ and $H_2O$, and the material balance equations for oxygen and carbon dioxide, oxygen uptake rate (OUR) rind carbon dioxide evolution rate (CER) were instantaneously calculated using a computer (16-bit IBM PC-AT) interfaced to a quadrupole mass spectrometer. The calculated OUR and CER data were used for the estimation of cell concentration and growth rate of Candida utilis during batch culture. It was found that the cell concentration could be satisfactorily estimated from the data of OUR arid CER during the culture and this method could be successfully und for the continuous monitoring of cell growth and growth rate.

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Electrodynamic Behavior of a Charged Particle among Two-Dimensional Quadrupole Electrodes (2차원 4극 전극 사이에서의 하전 입자의 동전기력학적 거동)

  • Park, Seok-Joo;Lim, Jeong-Hwan;Kim, Sang-Do;Choi, Ho-Kyung;Park, Hyun-Seol;Park, Young-Ok;McMurry, Peter-H.
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.25 no.5
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    • pp.741-749
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    • 2001
  • An inhomogeneous hyperbolic electric field is established among two-dimensional quadrupole electrodes to which an ac voltage is applied. Conditions under which charged particles are focused into a narrow axis region of the plug laminar flow are discussed. The aerodynamic forces influence the behavior of the charged particles in the quadrupole electric field. We derived the dimensionless equations of motion of a charged particle in the alternating quadrupole electric field, and discussed particle trajectories and focusing performance in terms of two dimensionless parameters, which are functions of particle size, operating pressure, and the amplitude and frequency of applied AC voltage, with the results of numerical simulations and experiments.

Design of an Air-Core HTS quadruple triplet for a heavy ion accelerator

  • Zhang, Zhan;Wei, Shaoqing;Lee, Sangjin
    • Progress in Superconductivity and Cryogenics
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    • v.18 no.4
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    • pp.35-39
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    • 2016
  • In recent years, high-temperature superconductor (HTS) Quadruple Triplets are being developed for heavy ion accelerators, because the HTS magnets are suitable to withstand radiation and high heat loads in the hot cell of accelerators. Generally, an iron yoke, which costs a mass of material, was employed to enhance the magnetic field when a quadrupole magnet was designed. The type of the magnet is called iron-dominated magnet, because the total magnetic field was mainly induced by the iron. However, in the HTS superconductor iron-dominated magnets, the coil-induced field also can have a certain proportion. Therefore, the air-core HTS quadrupole magnets can be considered instead of the iron-core HTS quadrupole magnet to be employed to save the iron material. This study presents the design of an air-core HTS quadruple triplet which consists three by air-core HTS quadruple magnet and compare the design result with that of an iron-core HTS quadruple triplet. First, the characteristics of an air-core HTS quadrupole magnet were analyzed to select the magnet system for the magnetic field uniformity impairment. Then, the field uniformity was improved(< 0.1%) exactly using evolution strategy (ES) method for each iron-core HTS quadrupole magnet and the air-core HTS quadruple triplet was established. Finally, the designed air-core triplet was compared with the iron-core HTS quadruple triplet, and the results of beam trajectories were presented with both the HTS quadruple triplet systems to show that the air-core triplet can be employed instead of the iron-core HTS triplet. The design of the air-core quadruple triplet was suggested for a heavy ion accelerator.

Continuous On-line Estimation of Cell Growth and Substrate Consumption Using a Computer-coupled Mass Spectrometer (Computer-coupled Mass Sepctrometer를 이용한 세포증식과 기질소모의 연속적 On-line추정)

  • 남수완;김정희
    • KSBB Journal
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    • v.4 no.2
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    • pp.118-122
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    • 1989
  • From the on-line mass spectrometric analyese of the exhaust gaseous composition of fermentor and the material balance equations for oxygen and carbon dioxide, oxygen uptake rate (OUR) and carbon dioxide evolution rate (CER) were calculate using a personal computer (IBM PC-AT) interfaced to a quadrupole mass spectromter. The calculate OUR and CER were used for the indirect estimation of cell and substrate concentrations during the batch culture of Candida utilis. For the estimation of sustrate concentration, the yield model of Pirt was applied. It was found that the cell and substrate (glucose) concentration could be ssatisfactorily estimataed and the results showed the more accurate estimations of both fermentation state variables when OUR data were used than CER data.

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Constructing Proteome Reference Map of the Porcine Jejunal Cell Line (IPEC-J2) by Label-Free Mass Spectrometry

  • Kim, Sang Hoon;Pajarillo, Edward Alain B.;Balolong, Marilen P.;Lee, Ji Yoon;Kang, Dae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1124-1131
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    • 2016
  • In this study, the global proteome of the IPEC-J2 cell line was evaluated using ultra-high performance liquid chromatography coupled to a quadrupole Q Exactive Orbitrap mass spectrometer. Proteins were isolated from highly confluent IPEC-J2 cells in biological replicates and analyzed by label-free mass spectrometry prior to matching against a porcine genomic dataset. The results identified 1,517 proteins, accounting for 7.35% of all genes in the porcine genome. The highly abundant proteins detected, such as actin, annexin A2, and AHNAK nucleoprotein, are involved in structural integrity, signaling mechanisms, and cellular homeostasis. The high abundance of heat shock proteins indicated their significance in cellular defenses, barrier function, and gut homeostasis. Pathway analysis and annotation using the Kyoto Encyclopedia of Genes and Genomes database resulted in a putative protein network map of the regulation of immunological responses and structural integrity in the cell line. The comprehensive proteome analysis of IPEC-J2 cells provides fundamental insights into overall protein expression and pathway dynamics that might be useful in cell adhesion studies and immunological applications.

Generation and Characterization of a Monoclonal Antibody with Specificity for Mycoplasma arginini

  • Son, Yeon-Sung;Hong, Hyo-Jeong
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.547-552
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    • 2007
  • Previously, we generated monoclonal antibodies (MAbs) that bound to the surface of human embryonic stem cells (hESCs) in an attempt to discover new hESC-specific surface markers. In this study, MAb 47-235 (IgG1, ${\kappa}$) was selected for further characterization. The MAb bound to the surface of undifferentiated hESCs but did not bind to mouse ESCs or mouse embryonic fibroblast cells in flow cytometric analysis. The antibody immunoprecipitated a 47 kDa protein from the lysates of cell surface-biotinylated hESCs. Identification of the protein by quadrupole time of flight tandem mass spectrometry revealed that 47-235 binds to Ag 243-5 protein of Mycoplasma arginini. BM-Cyclin treatment of the hESCs that reacted with 47-235 resulted in loss of mycoplasma DNA and the reactivity to 47-235. Nevertheless, the hESCs that were reactive to 47-235 maintained self-renewal and pluripotency and thus could be differentiated into three embryonic germ layers.

Metabolomics Investigation of Cutaneous T Cell Lymphoma Based on UHPLC-QTOF/MS

  • Zhou, Qing-Yuan;Wang, Yue-Lin;Li, Xia;Shen, Xiao-Yan;Li, Ke-Jia;Zheng, Jie;Yu, Yun-Qiu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.13
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    • pp.5417-5421
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    • 2014
  • Objectives: The identification of cutaneous T cell lymphoma (CTCL) biomarkers may serve as a predictor of disease progression and treatment response. The aim of this study was to map potential biomarkers in CTCL plasma. Design and Methods: Plasma metabolic perturbations between CTCL cases and healthy individuals were investigated using metabolomics and ultrahigh performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS). Results: Principal component analysis (PCA) of the spectra showed clear metabolic changes between the two groups. Thirty six potential biomarkers associated with CTCL were found. Conclusions: Based on PCA, several biomarkers were determined and further identified by LC/MS/MS analysis. All of these could be potential early markers of CTCL. In addition, we established that heparin as a nticoagulant has better pre-treatment results than EDTA with the UHPLC-QTOF/MS appraoch.

Comparison of Cell Lysis Techniques via Q-TOF LC/MS

  • Kaplan, Ozan;Oncul, Selin;Ercan, Ayse;Celebier, Mustafa
    • Mass Spectrometry Letters
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    • v.11 no.2
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    • pp.36-40
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    • 2020
  • Untargeted metabolomics is a useful tool for drug development focusing on novel chemotherapeutic and chemopreventative agents against cancer cells. In recent years, quadrupole time of flight liquid chromatography-mass spectrometry (Q-TOF LC/MS)-based untargeted metabolomic approaches have gained importance to evaluate the effect of these agents at the molecular level. The researchers working on cell culture studies still do not apply standardized methodologies on sample preparation for untargeted metabolomics approaches. In this study, the rough and wet lysis techniques performed on MCF-7 breast cancer cells were compared with each other via the Q-TOF LC/MS-based metabolomic approach. The C18 and hydrophilic interaction liquid chromatography (HILIC) columns were used for the separation of the metabolites in MCF-7 cell lysates. 505 peaks were detected through the HILIC column and 551 peaks were found through the C18 column for the wet lysis technique. This situation supported by the base peak chromatograms showed that the wet lysis technique allowed us to extract higher number of non-polar metabolites. Almost equal number of metabolites was found for the C18 and HILIC columns (697 peaks for the HILIC column and 695 peaks for the C18 column) when the rough lysis technique was used. However, the intensities of polar metabolites were higher for the rough lysis technique on base peak chromatograms for both the HILIC and C18 columns. Although cell lysis technique, which is the first step in the sample preparation for cell culture studies, does not cause dramatic differences in the number of the detected metabolite peaks, it affects the polar and non-polar metabolite ratio significantly. Therefore, it must be considered carefully especially for in vitro drug development studies.

Rhus verniciflua Stokes Extract and Its Flavonoids Protect PC-12 Cells against H2O2-Induced Cytotoxicity

  • Nam, Tae Gyu;Lee, Bong Han;Choi, Hyo-Kyoung;Mansur, Ahmad Rois;Lee, Sang Gil;Kim, Dae-Ok
    • Journal of Microbiology and Biotechnology
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    • v.27 no.6
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    • pp.1090-1097
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    • 2017
  • Rhus verniciflua Stokes (RVS), an herbal medicine found in East Asia, was extracted and further fractionated to investigate its antioxidant capacity and neuroprotective effects. The RVS ethyl acetate (EtOAc) fraction had the highest level of total phenolics and antioxidant capacity among all solvent fractions tested. Pretreatment of PC-12 cells with the EtOAc fraction effectively attenuated $H_2O_2$-induced oxidative damage. Furthermore, the EtOAc fraction significantly attenuated caspase-3 activity, resulting in inhibition of $H_2O_2$-induced apoptosis. We identified and quantified fustin, sulfuretin, and butein in the EtOAc fraction using accurate mass quadrupole time-of-flight mass spectrometry and reversed-phase high-performance liquid chromatography. The intracellular antioxidant capacity and superoxide dismutase (SOD) activity were significantly increased in PC-12 cells treated with the EtOAc fraction and with individual flavonoids. When cells were pretreated with the EtOAc fraction or individual flavonoids and then co-incubated with diethyldithiocarbamic acid (an inhibitor of SOD activity), cell viability against $H_2O_2$-induced oxidative stress was attenuated. These results suggest that the RVS EtOAc fraction and its flavonoid constituents protect PC-12 cells against $H_2O_2$-induced neurotoxicity through their antioxidant properties.

Profiling Analysis of Sphingolipids in HL-60 Cells by High-Performance Liquid Chromatography-Tandem Mass Spectrometry in combination with Multiple Reaction Monitoring

  • Son, Jung-Hyun;Lee, Jae-Ick;Yang, Ryung;Kim, Dong-Hyun
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.288.3-289
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    • 2003
  • Sphingolipid species are important second messengers due to their role in the mitogenesis, differentiation and apoptosis. We developed a new column liquid chromatography-triple quadrupole tandem mass spectrometry (LC-MS/MS) in combination with multiple reaction monitoring (MRM) method for the rapid, simultaneous and quantitative determination of unambiguous detecting sphingolipids in cell culture of human cancer cells (HL-60). (omitted)

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