• Asian Pacific Journal of Cancer Prevention
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• v.14 no.1
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• pp.367-372
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• 2013

Effects of the Epstein-Barr virus (EBV) on cellular protein expression are essential for viral pathogenesis. To characterize the cellular response to EBV infection, differential proteomes of gastric epithelial AGS cells were analyzed with two-dimensional gel electrophoresis (2-DE) followed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) and liquid chromatography electrospray/ionization ion trap (LC-ESI-IT) mass spectrometry identification. Mass spectrometry identified 9 altered cellular proteins, including 5 up-regulated and 4 down-regulated proteins after EBV infection. Notably 2-DE analysis revealed that EBV infection induced increased expression of heat shock cognate 71 kDa protein, actin cytoplasmic 1, pyridoxine-5'-phosphate oxidase, caspase 9, and t-complex protein 1 subunit alpha. In addition, EBV infection considerably suppressed those cellular proteins of zinc finger protein 2, cyclin-dependent kinase 2, macrophage-capping protein, and growth/differentiation factor 11. Furthermore, the differential expressional levels of partial proteins (cyclin-dependent kinase 2 and caspase 9) were confirmed by Western blot analysis.Thus, this work effectively provided useful protein-related information to facilitate further investigation of the mechanisms underlying EBV infection and pathogenesis.

• Bulletin of the Korean Chemical Society
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• v.31 no.8
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• pp.2293-2298
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• 2010

Focused electrospray (FES) deposition method is presented for matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. FES ion optics consists of two cylindrical focusing electrodes capped with a truncated conical electrode through which an electrospray emitter passes along the cylindrical axis. A spray of charged droplets is focused onto a sample well on a MALDI target plate under atmospheric pressure. The shape and size distributions of matrix crystals are visualized by scanning electron microscope and the mass spectra are obtained by time-of-flight mass spectrometry. Angiotensin II, bradykinin, and substance P are used as test samples, while $\alpha$-cyano-4-hydroxycinnamic acid and dihydroxybenzoic acid are employed as matrices. FES of a sample/matrix mixture produces fine crystal grains on a 1-3 mm spot and reproducibly yields the mass spectra with little shot-to-shot and spot-to-spot variations. Although FES greatly stabilizes the signals, the space charge due to matrix ions limits the detection sensitivity of peptides. To avoid the space charge problem, we adopted a dual FES/FES mode, which separately deposits matrix and sample by FES in sequence. The dual FES/FES mode reaches the detection sensitivity of 0.88 amol, enabling ultrasensitive of peptides by homogeneously depositing matrix and sample under atmospheric pressure.

• Journal of Radiation Protection and Research
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• v.10 no.1
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• pp.14-28
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• 1985

The development of mono energetic photon sources using $K_{\alpha}$ fluorescence X-ray of pure material was carried out in the energy range below 100 keV. The monoenergetic photons are very useful in the calibration of the radiation measuring instruments and can be produced as the $K_{\alpha}$ fluorescence X-ray by irradiating the bremsstrahlung to the thin pure metal foils called ‘radiators’. In this experiment, several radiators such as $_{47}Ag,\;_{50}Sn,\;_{68}Er,\;_{70}Yb,\;and\;_{82}Pb$ provide the wide monoenergetic photon energy ranging from 20 keV to 80 keV. By the spectrometry with HpGe LEPS, spectral purity factors which measure the monochrometicity for the $K_{\alpha}$ fluorescence X-ray, were determined as $0.64{\sim}0.94$. Dosimetry for the purpose of the determination of the exposure rate with a 600cc thin window ionization chamber, which was calibrated by the standard free-air ionization chamber, was performed. Exposure rates ranging $8.3{\sim}232.5mR/h$ was obtained according to the $K_{\alpha}$ fluorescence X-ray energy for each radiator.

• Journal of Microbiology and Biotechnology
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• v.24 no.2
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• pp.254-263
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• 2014

Enteromorpha polysaccharides (EP) extracted from green algae have displayed a wide variety of biological activities. However, their high molecular weight leads to a high viscosity and low solubility, and therefore, greatly restrains their application. To solve this problem, bacteria from the surface of Enteromorpha were screened, and an Alteromonas macleodii strain B7 was found to be able to decrease the molecular weight of EP in culture media. Proteins harvested from the supernatant of the A. macleodii B7 culture were subjected to native gel electrophoresis, and a band corresponding to the Enteromorpha polysaccharide lyase (EPL) was detected by activity staining. The enzyme identity was subsequently confirmed by MALDI-TOF/TOF mass spectrometry as the putative ${\alpha}$-amylase reported in A. macleodii ATCC 27126. The amylase gene (amySTU) from A. macleodii B7 was cloned into Escherichia coli, resulting in high-level expression of the recombinant enzyme with EP-degrading activity. AmySTU was found to be cold-adapted; however, its optimal enzyme activity was detected at $40^{\circ}C$. The ${\alpha}$-amylase was highly stable over a broad pH range (5.5-10) with the optimal pH at 7.5-8.0. The highest enzyme activity was detected when NaCl concentration was 2%, which dropped by 50% when the NaCl concentration was increased to 16%, showing an excellent nature of halotolerance. Furthermore, the amylase activity was not significantly affected by tested surfactants or the presence of some organic solvents. Therefore, the A. macleodii strain B7 and its ${\alpha}$-amylase can be useful in lowering EP molecular weight and in starch processing.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.9
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• pp.3849-3856
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• 2015

Background: Cholangiocarcinoma (CCA), or bile duct cancer, is incurable with a high mortality rate due to a lack of effective early diagnosis and treatment. Identifying cytoplasmic membrane proteins of invasive CCA that facilitate cancer progression would contribute toward the development of novel tumor markers and effective chemotherapy. Materials and Methods: An invasive CCA cell line (KKU-100) was stimulated using TNF-${\alpha}$ and then biotinylated and purified for mass spectrometry analysis. Novel proteins expressed were selected and their mRNAs expression levels were determined by real-time RT-PCR. In addition, the expression of ALCAM was selected for further observation by Western blot analysis, immunofluorescent imaging, and antibody neutralization assay. Results: After comparing the proteomics profile of TNF-${\alpha}$ induced invasive with non-treated control cells, over-expression of seven novel proteins was observed in the cytoplasmic membrane of TNF-${\alpha}$ stimulated CCA cells. Among these, ALCAM is a novel candidate which showed significant higher mRNA- and protein levels. Immunofluorescent assay also supported that ALCAM was expressed on the cell membrane of the cancer, with increasing intensity associated with TNF-${\alpha}$. Conclusions: This study indicated that ALCAM may be a novel protein candidate expressed on cytoplasmic membranes of invasive CCA cells that could be used as a biomarker for development of diagnosis, prognosis, and drug or antibody-based targeted therapies in the future.

• Journal of the Korean Chemical Society
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• v.17 no.1
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• pp.20-24
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• 1973

The average fission neutron cross sections were determined for the following reactions, $^{93}Nb(n,\alpha)^{90}Y$, $^{90}Zr(n,p)^{90}Y$,$^{93}Nb(n,\alpha)^{90m}Y$and$^{90}Zr(n,p)^{90m}Y$. The cation exchange column was used for the quantitative separation of the product nuclides using $\alpha-$hydroxyisobutyric acid as the eluent. The absolute activites of $^{90m}Y$ and $^{90}Y$were determined by the gamma ray spectrometry and a calibrated $2\pi$gas flow counter, respectively. The cross sections of $^{93}Nb(n,\alpha)^{90}Y$, $^{90}Zr(n,p)^{90}Y$,$^{93}Nb(n,\alpha)^{90m}Y$and $^{90}Zr(n,p)^{90m}Y$ reactions were found to be$0.14\pm0.01mb$, $0.83\pm0.02mb$, $0.018\pm0.02mb$ and $0.33\pm0.02mb$, respectively. The possible use of $^{90m}Y$ instead of $^{90}Y$ was discussed as a better means for the determination of niobium.

• Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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• v.18 no.2_spc
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• pp.227-235
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• 2020

In this study, we discussed the limitations of gross alpha measurements for the characterization of radioactive wastes produced in nuclear facilities through experimental tests and Monte Carlo N-particle transport simulations. The determination of gross alpha is essential for the disposal of radioactive waste produced in nuclear facilities in Korea. The measurements of gross alpha are easy to perform and yield rapid analytical results, but it cannot be used for quantitative analysis. The error of counting efficiency for gross alpha with various masses of the deposit on planchets using KCl and ²⁴¹Am was determined. The relative deviation of the counting efficiency in samples having the same mass was 20%. Uranium was extracted from the soil through acid leaching and extraction chromatography, and the concentration of U determined by inductively coupled plasma-mass spectrometry (ICP-MS) was compared with the results for gross alpha. The gross alpha was underestimated by 50% compared to the U concentration by ICP-MS. The counting efficiency depended on the energy from the alpha emitters, which differed by up to three times in determination of the counting efficiency depending on the kinds of alpha radionuclides of interest. Therefore, the gross alpha is not compatible with the sum of radioactivity for each alpha emitter and is suitable as a screening method.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.7
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• pp.3311-3317
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• 2014

Background: The consequence of Rho GDP dissociation inhibitor alpha (RhoGDI${\alpha}$) activity on migration and invasion of estrogen receptor positive ($ER^+$) and negative ($ER^-$) breast cancer cells has not been studied using the proteomic approach. Changes in expression of RhoGDI${\alpha}$ and other proteins interacting directly or indirectly with RhoGDI${\alpha}$ in MCF7 and MDA-MB-231, with different metastatic potentials is of particular interest. Materials and Methods: $ER^+$ MCF7 and ER- MDA-MB-231 cell lines were subjected to two-dimensional electrophoresis (2-DE) and spots of interest were identified by matrix-assisted laser desorption/ionization time of- flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry (MS) analysis after downregulation of RhoGDI${\alpha}$ using short interfering RNA (siRNA) and upregulated using GFP-tagged ORF clone of RhoGDI${\alpha}$. Results: The results showed a total of 35 proteins that were either up- or down-regulated in these cells. Here we identifed 9 and 15 proteins differentially expressed with silencing of RhoGDI${\alpha}$ in MCF-7 and the MDA-MB-231 cells, respectively. In addition, 10 proteins were differentially expressed in the upregulation of RhoGDI${\alpha}$ in MCF7, while only one protein was identified in the upregulation of RhoGDI${\alpha}$ in MDA-MB-231. Based on the biological functions of these proteins, the results revealed that proteins involved in cell migration are more strongly altered with RhoGDI-${\alpha}$ activity. Although several of these proteins have been previously indicated in tumorigenesis and invasiveness of breast cancer cells, some ohave not been previously reported to be involved in breast cancer migration. Hence, these proteins may serve as useful candidate biomarkers for tumorigenesis and invasiveness of breast cancer cells. Conclusions: Future studies are needed to determine the mechanisms by which these proteins regulate cell migration. The combination of RhoGDI${\alpha}$ with other potential biomarkers may be a more promising approach in the inhibition of breast cancer cell migration.

• Food Science and Preservation
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• v.23 no.7
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• pp.977-988
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• 2016

The volatile flavor compounds in five Jeju citrus fruit varieties (Cheonhyehyang, Hallabong, Jinjihyang, Hwanggeum hyang and Redhyang) were extracted by simultaneous distillation-extraction (SDE) using solvent mixture of n-pentane and diethyl ether (1:1, v/v) and analyzed by using gas chromatography-mass spectrometry (GC-MS). The number of aroma compounds were identified with : 104 (4,939.77 mg/kg) in Cheonhyehyang, 98 (3,286.38 mg/kg) in Hallabong, 105 (3,317.56 mg/kg) in Jinjihyang, 102 (4,293.39 mg/kg) in Hwanggeumhyang, and 108 (4,049.94 mg/kg) in Redhyang. The detected main volatile compounds were; limonene, sabinene, ${\beta}$-myrcene, ${\alpha}$-pinene, ${\beta}$-pinene, linalool, 4-terpineol, ${\alpha}$-terpineol, (E)-${\beta}$-ocimene and ${\gamma}$-terpinene. Among the identified volatiles compounds, ethyl-benzene, nonanol, 1-p-menthen-9-al, (E)-isocarveol, methyl salicylate, ${\alpha}$-terpinen-7-al, perilla alcohol, and ethyl-dodecanoate were detected in Cheonhyehyang. only Furthermore, ${\beta}$-chamigrene and ${\alpha}$-selinene were in Hallabong only; 3-hydroxybutanal, (E)-2-nonenal, isoborneol, octyl acetate, (E)-2-undecenal, ${\beta}$-ylangene and guaia-6,9-diene in Jinjihyang. ${\rho}$-Cymenene, ${\beta}$-thujone, selina-4,11-diene and (E,E)-2,6-farnesol in Hwanggeumhyang only; and ${\rho}$-cymen-8-ol, bornyl acetate, carvacrol, bicycloelemene, ${\alpha}$-cubebene and 7-epi-${\alpha}$-selinene in Redhyang only. This study confirmed the differences in composition and content of volatile aroma components in five varieties of Jeju citrus fruits.

• Journal of the Korean Society of Food Culture
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• v.24 no.1
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• pp.84-89
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• 2009

This paper was conducted to evaluate aglycones and carbohydrates produced by acid hydrolysis of three potato glycoalkaloids [(PGA); ${\alpha}$-chaconine, ${\alpha}$-solanine, and demissine] in potatoes. Standard solanidine and demissidine were dissolved in 1N HCl and then heated at $100^{\circ}C$ for 10-120 min. Solanidine was rapidly decomposed during acid hydrolysis and one peak that was identified as solantherene ($M^+$=379) by GC-MS was detected. The transformation solanidine to solanthrene was approximately 50% complete after 10 min, approximately 90% complete after 60 min and 100% complete after 120 min. Demissidine was hydrolyzed using the same method that was used to hydrolyze the solanidine. However, demissidine produced only one peak upon GC-MS ($M^+$=399) analysis and was found to be very stable at increased temperatures. Acidy hydrolysis of ${\alpha}$-chaconine, ${\alpha}$-solanine and demissine resulted in the decomposition of ${\alpha}$-chaconine and ${\alpha}$-solanine to solanidine and solanthrene, respectively. Therefore, this hydrolysis method should not be utilized to produce PGA combining with solanidine as aglycone. The individual carbohydrates produced by the two PGAs by hydrolysis were very stable at increased temperatures; therefore, it was possible to quantify these PGAs based on calculation of the individual carbohydrate content. Conversely, because demissidine produced by the hydrolysis of demissine was extremely stable at increased temperatures, it was possible to quantify the PGA based on the aglycone produced by hydrolysis.