• Journal of Life Science
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• v.27 no.8
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• pp.864-872
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• 2017

Equine chorionic gonadotropin (eCG) consists of highly glycosylated ${\alpha}-$ and ${\beta}-subunits$ and is a unique member of the gonadotropin family, because it elicits the response characteristics of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in species other than the horse. To directly assess the biological function of $rec-eCG{\beta}/{\alpha}$, we constructed mammalian expressing vectors of equine luteinizing hormone/chorionic gonadotropin receptors (eLH/CGR). The activity of $rec-eCG{\beta}/{\alpha}$ in vitro assayed in transient transfected CHO-K1 cells and in stably transfected PathHunter Parental cells with eLH/CGR was investigated. $rec-eCG{\beta}/{\alpha}$ was efficiently secreted in the CHO-K1 suspension cell media, and the quantity detected was about 200 mIU/ml from 1 to 7 days after transfection. In the western blot analysis, the $rec-eCG{\beta}/{\alpha}$ protein was broadly identified to be about 40~45 kDa molecular weight. The cAMP stimulation in CHO-K1 cells expressing eLH/CGR was determined to evaluate the activity of $rec-eCG{\beta}/{\alpha}$. The cAMP concentration increased in direct proportion to the concentration of the $rec-eCG{\beta}/{\alpha}$. The $EC_{50}$ value in the transient transfected CHO-K1 cells was $8.1{\pm}6.5ng$. The stable cell lines of eLH/CGR were established in the PathHunter Parental cells expressing ${\beta}-arrestin$. We found that $rec-eCG{\beta}/{\alpha}$ had full LH activity in the PathHunter Parental cells expressing eLH/CGR. The $EC_{50}$ value in transient and stable cells was $5.0{\pm}4.7ng/ml$ and $4.5{\pm}5.2ng/ml$, respectively. These results suggest that $rec-eCG{\beta}/{\alpha}$ has a biological activity in a cell expressing eLH/CGR. These stable cells expressed in PathHunter Parental cells could be useful for elucidating the functional mechanisms of deglycosylated $rec-eCG{\beta}/{\alpha}$ mutants.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.6
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• pp.767-774
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• 2011

Defatted green tea seed was extracted with 100% ethanol for 4 hr and then fractionated with petroleum ether, ethyl acetate and butanol. The ethanol and butanol extracts showed greater increases in antiproliferation potential against liver cancer cells than petroleum ether, ethyl acetate, $H_2O$, and hot water extracts did. Thus, this study was carried out to investigate the anti-proliferative actions of defatted green tea seed ethanol extract (DGTSE) in HepG2 cancer cells. The DGTSE contained catechins including EGC ($1039.1{\pm}15.2\;g/g$), tannic acid ($683.5{\pm}17.61\;{\mu}g/g$), EC ($62.4{\pm}5.00\;{\mu}g/g$), ECG ($24.4{\pm}7.81\;{\mu}g/g$), EGCG ($20.9{\pm}0.96\;{\mu}g/g$) and gallic acid ($2.4{\pm}0.68\;{\mu}g/g$), but caffeic acid was not detected when analyzed by HPLC. The anti-proliferation effect of DGTSE toward HepG2 cells was 83.13% when treated at $10\;{\mu}g$/mL, of DGTSE, offering an $IC_{50}$ of $6.58\;{\mu}g$/mL. DGTSE decreased CYP1A1 and CYP1A2 protein expressions in a dose-dependent manner. Quinone reductase and antioxidant response element (ARE)-luciferase activities were increased about 2.6 and 1.94-fold at a concentration of $20\;{\mu}g$/mL compared to a control group, respectively. Enhancement of phase II enzyme activity by DGTSE was shown to be mediated via interaction with ARE sequences in genes encoding the phase enzymes. DGTSE significantly (p<0.05) suppressed prostaglandin $E_2$ level, tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) protein expressions, and NF${\kappa}$B translocation, but did not affected nitric oxide production. From the above results, it is concluded that DGTSE may ameliorate tumor and inflammatory reactions through the elevation of phase II enzyme activities and suppression of NF${\kappa}$B translocation and TNF-${\alpha}$ protein expressions, which support the cancer cell anti-proliferative effects of DGTSE in HepG2 cells.

• Clinical and Experimental Pediatrics
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• v.46 no.5
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• pp.467-473
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• 2003

Purpose : The Epstein-Barr virus(EBV), gamma herpesvirus, is an important pathogen that is widespread around the world. The EBV causes various diseases depending on the geographic location, and on the immunity or the premorbid condition of the person exposed to EBV. To evaluate EBV typing may be the most important step to figure out the pathogenesis of EBV associated diseases, and we need to re-evaluate the pathologic role of human leukocyte antigen(HLA) in developing Epstein- Barr virus associated acute infectious mononucleosis by using newly developed methods. Methods : This study included 24 children(age range : 6 to 13 years), serologically confirmed with acute infectious mononucleosis. The control group for the HLA type consisted of 200 age-matched healthy children. To classify HLA I, modified ARMs-PCR was used, while modified PCR-SSOP was utilized in typing of HLA II. Also, we performed EBV typing in study patients by using a one-step PCR. Results : The results of HLA types : In HLA class I, HLA-A24 was positive in 69 of 200 healthy children and positive in 14 of 24 patients in the study group(relative risk : 3.5724, chi-square; 5.26, P<0.05). In HLA class II, HLA-DRB1*07 was detected in 18 of 200 healthy children, and eight of 24 patients in the study group(relative risk; 506173, chi-square; 9.73, P<0.01). The results of EBV types : In the research group, 20(83.8%) of 24 patients were shedding type A virus, while 4(16.7%) were type B. Conclusion : We conclude that development of infectious mononucleosis may be associated with HLA types, and these results suggest that acute infectious mononucleosis could have hereditary traits. And we confirm that type A EBV is highly prevalent in patients with acute infectious mononucleosis in Korea. Also, our results suggest that further large scale studies, including adult groups, regarding the association between pathogenesis of EBV with HLA-DP or HLA-DQ will be warranted.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.412-420
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• 1989

To know how the ribosomes involved in secretory protein synthesis were attached to the cytoplasmic membrane in Bacillus amyloliquefaciens, the cells were treated with puromycin combinated with magnesium at the logarithmic phase, and the variation of cell-bound and extracellular $\alpha$-amylase activity was assayed for determining the $\alpha$-amylase translocation blocking through the cytoplasmic membrane. In the abnormal $\alpha$-amylase producing mutant in which the C-terminal of the $\alpha$-amylase structure was deleted, B. umytotiquefaciens CH10-2, the $\alpha$-amylase was translocated normally through the cytoplasmic membranes, and the translocation blocking by puromycin was revealed to have a similar pattern as that in the wild type. This means that the C-terminal part of the enzyme structure may not have a signal for secretion. The cell death of the logarithmic phase cells in both strains was not affected much under 20$\mu\textrm{g}$/$m\ell$ of puromycin, however, the $\alpha$-amylase translocation was blocked markedly under less than 10$\mu\textrm{g}$/$m\ell$ of the puromycin concentration. The blocking of the enzyme secretion by puromycin may be due to the detachment of the ribosomes from cytoplasmic membranes by disturbing the nascent polypeptide synthesis. Further evidence for confirming this was that the detachment was increased in 50 mM of magnesium ion because the extracellular $\alpha$-amylase activity was decreased more under this condition. If the cells were treated with trypsin combinated with Iysozyme, the extracellular $\alpha$-amylase activity from the cultured medium was reduced markedly, however, the activity from the cells treated with trypsin only was not reduced. This means that the nascent polypeptides protruding from the cytoplasmic membrane were sensitive to the trypsin digestion, whereas the matured ones were not. Therefore, the protruding polypeptides from the cytoplasmic membranes may be truncated by trypsin before forming their final tertiary structures by folding in the cell wall layer.

• The Korean Journal of Pesticide Science
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• v.18 no.2
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• pp.79-87
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• 2014

Pyriofenone is an aryl phenyl ketone fungicide that is newly registered in Korea in 2013 to control powdery mildew on food. The objective of this study was to develop reliable and sensitive analytical method for determination of pyriofenone residue in agricultural products for ensuring the food safety. The pyriofenone residues in all samples(Korean melon, pepper, potato, mandarin, soybean, and hulled rice) were extracted with acetonitrile, partitioned with dichloromethane, and then purified with a silica cartridge. The purified samples were analyzed by HPLC-UVD and confirmed with LC-MS. The linear range of pyriofenone was 0.05~5 mg/kg with the correlation coefficient ($r^2$) > 0.999. Average recoveries of pyriofenone ranged from 72.8% to 99.5% at the spiked level of 0.05 and 0.5 mg/kg, while the relative standard deviation was 2.3%~6.4%. In addition, the limit of detection and limit of quantification were 0.01 and 0.05 mg/kg, respectively. The results revealed that the developed and validated analytical method was suitable for pyriofenone determination in agricultural products.

• Korean Journal of Medicinal Crop Science
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• v.3 no.3
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• pp.217-225
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• 1995

In this study the essential oil and pharmacologically active components of different valerian species (Valeriana fauriei var. dasycarpa Hara and V. officinalis L. from Korea and V. wallichii DC from Nepal) were analyzed to search for native plant resources that can be used for medicinal herb and tobacco flavoring. The oil contents in dry roots of V. fauriei, V. officinalis and V. walhchii were 1. 71%, 0. 82% and 0. 83 %, respectively. The composition of the essential oil among different valerian species was compared. In V. fauriei 47 compounds were identified, while 21 and 25 compounds were identified in V. officinalis and V. wallichii. The major compounds in the oil of V. fauriei were bornyl acetate (33.8%) and camphene (10.8%), cedrol (4. 1 %), -pinene (3. 2%) and unidentified sesquiterpene alcohol (3. 0%). The major compounds were borneol (62. 5%) and ${\beta}-sesquiphelandrene$(6. 8%) and spathulenol (2. 1%) in V. officinalis, and borneol (74. 3%) and ${\alpha}-elemene$ (2.7%) and -sesquiphellandrene (2. 3%) in V. wallichii. Among the components known as major pharmacologically active compounds in European or Japanese valerian roots, valeranone, valerenal and -kessyl acetate was detected in a small amount in V. fauriei, but kessoglycol diacetate was not detected in V. faudei and V. officinalis Among the valepotrate compounds, major pharmacologically active components in V. wallichii, valtrate was detected in a small amount in V. fauriei and V. officinalis., and detected 1. 42% in V. wallichii. Didrovaltrate was also detected in the three valerian species tested, but acevaltrate was not detected except V. wallichii. On the other hand, antioxidative activity was slightly higher in V. fauriei than those of V. officinalis.

• Journal of Mushroom
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• v.13 no.4
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• pp.319-325
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• 2015

This study was carried out for examining the amount of ganoderic acid A, F, H from fruiting body extracts of the various Ganoderma species. The preserved Ganoderma species are extracted by using three kinds of solvent. Among them, five strains which have the large amount of ganoderic acid A, F, H were selected. First of all, Strains with the highest amount of ganoderic acid A were sorted out acorrding to solvent as follows. ASI 7013 has the highest amounts with 0.988 mg/g from D.W. extracts. And ASI 7023, 7059, 7026, 7060 were in order of content. In EtOH extracts, ASI 7037 has the highest amounts of ganoderic acid A with 0.940 mg/g. And ASI 7021, 7034, 7026, 7038 were in order of content. In MeOH extracts, ASI 7162 has the highest amounts with 0.833 mg/g. And ASI 7091, 7032, 7056, 7125 were in order of content. secondly, Strains with the highest amount of ganoderic acid F were sorted out acorrding to solvent as follows. ASI 7011 has the highest amounts with 0.722 mg/g from D.W. extracts. And ASI 7113, 7011, 7104 were in order of content. In EtoH extracts, ASI 7037 has the highest amounts with 0.811 mg/g. And ASI 7007, 7027, 7012, 7014 were in order of content. In MeoH extracts, ASI 7162 has the highest amounts with 0.833 mg/g. And ASI 7021, 7034, 7026, 7125 were in order of content. Finally, Strains with the highest amount of ganoderic acid H were sorted out acorrding to solvent as follows. ASI 7013 has the highest amounts with 0.985 mg/g in D.W. extracts. And ASI 7059, 7135, 7001, 7141 were in order of content. In EtoH extracts, ASI 7016 has the highest amounts with 2.842 mg/g. And ASI 7089, 7159, 7007, 7002 were in order of content. In MeoH extracts, ASI 7003 has the highest amounts with 6.969 mg/g. And ASI 7034, 7169, 7159, 7103 were in order of content.

• Journal of Food Hygiene and Safety
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• v.26 no.2
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• pp.142-149
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• 2011

An isocratic high performance liquid chromatography (HPLC) method for routine analysis of deoxynivalenol in noodles was validated and estimated the measurement uncertainty. Noodles (dried noodle and ramyeon) were analyzed by HPLC-ultraviolet detection using immunoaffinity column for clean-up. The limits of detection (LOD) and quantification (LOQ) were 7.5 ${\mu}g$/kg and 18.8 ${\mu}g$/kg, respectively. The calibration curve showed a good linearity, with correlation coefficients $r^2$ of 0.9999 in the concentration range from 20 to 500 ${\mu}g$/kg. Recoveries and Repeatabilities expressed as coefficients of variation (CV) spiked with 200 and 500 ${\mu}g$/kg were $82{\pm}2.7%$ and $87{\pm}1.3%$% in dried noodle, and $97{\pm}1.6%$ and $91{\pm}12.0%$ in ramyeon, respectively. The uncertainty sources in measurement process were identified as sample weight, final volume, and sample concentration in extraction volume as well as components such as standard stock solution, working standard solution, 5 standard solutions, calibration curve, matrix, and instrument. Deoxynivalenol concentration and expanded uncertainty in two matrixes spiked with 200 ${\mu}g$/kg and 500 ${\mu}g$/kg were estimated to be $163.8{\pm}52.1$ and $435.2{\pm}91.6\;{\mu}g$/kg for dried noodle, and $194.3{\pm}33.0$ and $453.2{\pm}91.1\;{\mu}g$/kg for ramyeon using a coverage factor of two which gives a level of statistical confidence with approximately 95%. The most influential component among uncertainty sources was the recovery of matrix, followed by calibration curve.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.6
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• pp.693-698
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• 1999

To investigate changes of components in salt-fermented northern sand lance, Ammodytes personatus sauce during fermentation, various chemical properties were examined at 1$\~$3 months intervals during 18 months fermentation. The moisture content decreased slightly, but the content of VBN and crude protein, total nitrogen, amino nitrogen, degree of hydrolysis, and absorbance at 453 nm increased gradually during fermentation. On the other hand, ash content, pH, and salinity showed almost no change. The contents of total nitrogen, amino nitrogen, and degree of hydrolysis increased sharply until 6$\~$8 months fermentation and showed the gentle increment after that, The Hx and uric acid were the most abundant in ATP related compounds, ranging from $83.1\%$ to $92.9\%$, After 18 month of fermentation, sauce was rich in free amino acids, such as glutamic acid, alanine, Iysine, leucine, isoleucine, valine, aspartic acid in that order.

• Journal of Chest Surgery
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• v.35 no.2
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• pp.94-101
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• 2002

All kinds of tissue valves must be pretreated for the inactivation of immunologic properties and the strengthening of tissue before implantation. However, the tissue valves are gradually denatured with the calcification process and they eventually lose their functions. Recent reports have shown the existence of specific calcium binding non collagenous proteins in the calcified area of implanted biomaterials. This experiment was intended to confirm the effect of pretreatment with RGDS(Arg-Gly-Asp-Ser) tetrapeptide on the calcification of subcutaneously implanted bovine pericardium in rats. RGDS tetrapeptide has the same amino acid sequence of attachment site of specific calcium binding non collagenous proteins. Material and Method: All bovine pericardial pieces were fixed with 0.6% glutaraldehyde. The pretreatments were done using 5 different methods, groupI, with normal saline for 60 minutes, groupII, with 0.5% GRSD(Gly-Arg-Scr-Asp) tetrapeptide solution for 60 minutes, group III : with 0.5% RGDS(Arg-Gly-Asp-Ser) tctrapeptide for 30 minutes, group IV ; with 0.5% RGDS for 60 minutes, and group V : with 0.5% RGDS for 120 minutes. The pretreated bovine pericardial pieces were implanted subcutaneously at the abdominal sites of rats. 30 days after the implantation, the implanted bovine pericardial tissue were examined radiologically, biochemically, and histologically to measure the severity of calcification. Result: On the radiological examination, group I ; 68.42$\pm$3.06, group II , 64.25$\pm$5.58 showed significant difference with group III: 48.00$\pm$3.57, group IV; 43.67$\pm$2.31, and group V ; 2.58$\pm$2.47(p<0.05). There was no difference between group I and II(p=0.105). On the biochemical examination, the amount of calcium in group I was , 33.09$\pm$6.59 mg, in group II ; 28.12$\pm$5.50mg, in group III ; 25.42$\pm$7.67mg, in group Ⅵ ; 20.51$\pm$5.11mg, and in group V : 15.43$\pm$4.25mg.