• Parasites, Hosts and Diseases
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• v.29 no.3
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• pp.293-310
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• 1991

Clonorchis sinensis is a common parasite of man in Korea. Researches on the specific antigens of C. sinensis would be valuable not only because those elucidate the molecular characteristics of this fluke but also because it is applicable to immunodiagnosis. Although many monoclonal antibodies have been used in the field of parasite immunology, few articles on monoclonal antibodies against C. sinensis have been published so far. The aim of this study was to analyse C. sinensis antigens recognized by monoclonal antibodies, and to set up ELISA-inhibition test using C. sinensis specific monoclonal antibodies for improved specificity of immunodiagnostic tests. By fusion between spleen cells of the mice immunized with C. sinensis water-soluble crude adult worm antigens and plasmacytoma cells of mouse origin, 29 hybridoma clones secreting anti-C. sinensis monoclonal antibodies were made, and 8 clones among those were found specific. After cell cloning, isotypes of 6 selected specific monoclonal anti- bodies were determined to be IgGl, IgG2b and IgA. Four exposed antigenic determinants of natural infection were recognized by different specific monoclonal antibodies. By enzyme-immunoelectrotransfer blot, 10 KD, 34 KD antigenic determinants were found to be reacted with CsHyb 0714-20, CsHyb 0605-10 monoclonal antibodies, respectively, The antigenic determinant recognized by CsHyb 0714-20 monoclonal antibody was revealed to be located at the surface and parenchyme of a parasite by indirect immunoauorescent antibody technique, and those reacted with CsHyb 0605-10, CsHyb 0714-25 monoclonal antibodies were found at the parenchyme and intestine. The antigenic determinant reacted with CsHyb 0605-23 monoclonal antibody was found mainly around the uterine eggs. Four antigenic determinants recognized by specific monoclonal antibodies were all found to be present in the early eluted fractions of C. sinensis antigens separated by Sephadex G-200 gel filtration. By conventional ELISA, 75% of clonorchiasis cases were found positive, but 7.1% of normal controls and 37.5% of paragonimiasis cases showed false positives. However, by ELISA-inhibition test using C. sinensis specific monoclonal antibody (CsHyb 0605-23), 77.1% of clonorchiasis cases were found positive, and there were no false positives in normal controls or paragonimiasis cases, indicating 100% specificity. The ELISA- inhibition test using monoclonal antibodies was found to have same sensitivity and definitely high specificity in comparison with conventional ELISA for serodiagnosis of human clonorchiasis.

• Journal of Genetic Medicine
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• v.7 no.2
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• pp.138-144
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• 2010

Purpose: Endothelial progenitor cells (EPCs), which mediates neovascularization of uterine endometrium may be involved in the neovascularization in the utero-placental circulation. Low numbers of endothelial progenitor colony-forming unit (CFU) in culture are predictive biomarker of vascular disease. The aim of the present study was to evaluate whether the number of CFU in preeclampsia differed from that in normal pregnancy. Materials and Methods: Women with singleton normal (n=26) or preeclamptic (n=20) pregnancies were studied during the third trimester. The number of EPCs was quantified by CFU methodology. Plasma levels of angiogenic factors, vascular endothelial growth factor (VEGF), soluble fms-like tyrosine kinase-1 (sFlt-1), and placental growth factor (PlGF) were determined by enzyme-linked immunoassay. Results: CFU numbers were significantly decreased in the preeclamptic patients compared with the controls (median, 3; range 1-12 vs. 31; 3-81 CFU/well, P<0.001). A majority of the cells comprising individual colonies were positive for endothelial characteristics (Ulex europaeus lectin staining and acetylated low-density lipoprotein uptake). Plasma levels of the sFlt-1 were highly elevated (P<0.001) in patient with preeclampsia compared to controls, whereas PlGF were highly reduced (P=0.004), but these factors did not associate with CFU numbers. Conclusion: Our results suggest that reduced numbers of CFU obtained from maternal peripheral blood may contribute to the development of preeclampsia.

• Horticultural Science & Technology
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• v.31 no.1
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• pp.80-88
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• 2013

Carrot (Daucus carota L. var. sativa) is one of the most extensively used vegetable crops in the world and a significant source of nutrient because of its high content of ${\beta}$-carotene, well known as the precursor of vitamin A carotenoid. However, seed-hairs generated and elongated from the epidermal cell of seeds inhibit absorption and germination by various factors such as carotol and so on. Accordingly, mechanical hair removal process is essential before commercialization of carrot seeds. Because of this process, producers will have additional losses such as time consuming, manpower, capital and so on. Furthermore, physical damage of seeds causes irregular germination rate. To overcome such cumbersome weaknesses, new breeding program for developing hairless-seed carrot cultivar has been needed and studies for molecular markers related to seed-hair characteristic is needed for a new breeding program. Therefore, in this study, cDNA libraries from seeds of short-hair seed phenotype CT-SMR 616 OP 659-1 line, hairy-seed phenotype CT-SMR 616 OP 677-14 line and short-hair seed phenotype CT-ATR 615 OP 666-13 line, hairy-seed phenotype CT-ATR 615 OP 671-9 were constructed, respectively. Furthermore, 1,248 ESTs in each line, total 4,992 ESTs were sequenced. As a result, 19 SNP sites and 14 SNP sites in each of 2 combinations were confirmed by analyzing these EST sequences from short-hair and hairy-seed lines. Then we designed SNP primer sets from EST sequences of SNP sites for high resolution melting (HRM) analysis. Designed HRM primers were analyzed using hairy seed phenotype CT-SMR 616 OP 1040 line and short-hair seed phenotype CT-SMR 616 OP 1024, 1025, 1026 lines. One set of HRM primers showed specific difference between the melting curves of hairy and short-hair seed phenotype lines. Based on this result, allele-specific (AS) PCR primers were designed for easier selection between hairy-seed carrot and hairless seed carrot. These results of HRM and AS-PCR are expected to be useful in breeding of hairless seed carrot cultivar as a molecular marker.

• Korean Journal of Food Science and Technology
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• v.46 no.3
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• pp.384-389
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• 2014

The objective of the present study was to investigate the chemical composition of anthocyanin-enriched extract of radiation-induced blackberry (Rubus fruticosus L.) mutant (${\gamma}$-B201) as well as the protective effect of ${\gamma}$-B201 against oxidative stress in vitro. The cytotoxicity, reactive oxygen species (ROS) scavenging capacity, and DNA damage were assessed by WST-1 assay, flow cytometry, and comet assay, respectively. Lactate dehydrogenase, superoxide dismutase, and catalase activities were determined by using a commercial kit. The in vitro results showed that ${\gamma}$-B201 increased the cell viability, reduction of lactate dehydrogenase release, and intracellular ROS scavenging capacity in hydrogen peroxide ($H_2O_2$)-treated HepG2 cells. Furthermore, treatment with ${\gamma}$-B201 attenuated DNA damage in $H_2O_2$-treated HepG2 cells and treatment with ${\gamma}$-B201 restored the activity of superoxide dismutase and catalase in $H_2O_2$-treated HepG2 cells. In conclusion, the present study suggests that ${\gamma}$-B201 blackberry extract can exert a significant cytoprotective effect against $H_2O_2$-induced cell damage.

• Korean Journal of Food Science and Technology
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• v.48 no.5
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• pp.508-514
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• 2016

Effects of the ethyl acetate fraction from gomchwi (Ligularia fischeri) extract against high $glucose/H_2O_2-induced$ oxidative stress and in vitro neurodegeneration were investigated to confirm the physiological property of the extract. The ethyl acetate fraction of gomchwi extract showed the highest total phenolic contents than the other solvent fractions. An anti-hyperglycemic activity of the ethyl acetate fraction was evaluated using the ${\alpha}-glucosidase$ inhibitory assay, and the half maximal inhibitory concentration ($IC_{50}$) value for ${\alpha}-glucosidase$ was found to be $727.64{\mu}g/mL$. In addition, the ethyl acetate fraction showed excellent 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt radical scavenging activity, and inhibition of malondialdehyde production. The ethyl acetate fraction also decreased intracellular reactive oxygen species, whereas neuronal cell viability against high glucose/$H_2O_2$-induced cytotoxicity was found to be increased. Finally, 3,5-dicaffeoylquinic acid as a main phenolic compound in the ethyl acetate fraction was analyzed by high-performance liquid chromatography. These results suggest that gomchwi might be a good natural source of functional materials to prevent diabetic neurodegeneration.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.9
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• pp.1159-1165
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• 2006

Our preliminary study showed that genistein and daidzein improved blood glucose level in type 2 diabetic mice by enhancing the glucose and lipid metabolism. The objective of this study was to evaluate whether genistein and daidzein are associated with alterations in antioxidant defense mechanism of type 2 diabetic mice. Male C57BL/KsJ-db/db (db/db) mice and age-matched non-diabetic littermates (db/+) were used in this study. The db/db mice were divided into control, genistein (0.02%, w/w) and daidzein (0.02%, w/w) groups. The relative weights of liver, epididymal adipose tissue and perirenal adipose tissue were significantly higher in the db/db group than in the db/+ group, whereas heart weight was lower. The genistein and daidzein supplement did not affect the organ weights in db/db mice. The blood glucose level was positively correlated with superoxide dismutase (SOD, r=0.380, p<0.05) and catalase (CAT, r=0.345, p<0.05) activities and negatively correlated with glutathione peroxidase (GSH Px, r= 0.404, p<0.05) activity in erythrocyte. Therefore, the erythrocyte SOD and CAT activities were significantly elevated in the db/db group compared to the db/+ group and the GSH-Px activity was lowered. However, the supplementation of genistein and daidzein reversed erythrocyte CAT and GSH-Px activities in type 2 diabetic mice. In this current study, the SOD activities in liver, kidney and heart were significantly not different between the groups. The CAT and GSH-Px activities in liver and GSH-Px activity in kidney were significantly higher in the db/db group than in the db/+ group, while the CAT activity in kidney, CAT and GSH-Px activities in heart were lowered. The supplementation of genistein and daidzein significantly attenuated the changes of CAT and/or GSH-Px activities in liver and heart. The supplementation of genistein and daidzein elevated GSH levels in kidney and heart compared to the db/db control group. The lipid peroxide levels in liver, kidney and heart were significantly lowered in the genistein and daidzein supplemented groups compared to the db/db control group. These results suggest that genistein and daidzein might be beneficial for the prevention of type 2 diabetic complication via suppressing changes of antioxidant enzymes activities with simultaneous reduction of lipid peroxidation.

• Journal of Life Science
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• v.29 no.4
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• pp.410-420
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• 2019

Citrus unshiu peel extracts possess a variety of beneficial effects, and studies on their anticancer activity have been reported. However, the exact mechanisms underlying this activity remain unclear. In the current study, the apoptotic effect of ethanol extract of C. unshiu peel (EECU) on human breast adenocarcinoma MDA-MB-231 cells and related mechanisms were investigated. The results showed that the survival rate of MDA-MB-231 cells treated with EECU was significantly inhibited in a concentration-dependent manner, which was associated with the induction of apoptosis. EECU-induced apoptosis was associated with the activation of caspase-8 and caspase-9, which initiate extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3, a representative effect caspase. EECU suppressed the expression of the inhibitor of apoptosis family of proteins, leading to an increased Bax/Bcl-2 ratio and proteolytic degradation of poly (ADP-ribose) polymerase. EECU also enhanced the loss of the mitochondrial membrane potential and cytochrome c release from the mitochondria to the cytosol, along with truncation of Bid. In addition, EECU activated AMP-activated protein kinase (AMPK), and compound C, an AMPK inhibitor, significantly weakened EECU-induced apoptosis and cell viability reduction. Furthermore, EECU promoted the generation of reactive oxygen species (ROS), which acted as upstream signals for AMPK activation as pretreatment of cells, with the antioxidant N-acetyl cysteine reversing both EECU-induced AMPK activation and apoptosis. Collectively, these findings suggest that EECU inhibits MDA-MB-231 adenocarcinoma cell proliferation by activating intrinsic and extrinsic apoptotic pathways, which was mediated through ROS/AMPK-dependent pathways.

• Journal of Life Science
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• v.28 no.11
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• pp.1347-1353
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• 2018

The fermentation of non-digestible soy meal can convert polysaccharides into many compounds that have a wide variety of biological functions. Bacillus strains are capable of hydrolyzing non-digestible saccharides, such as melibiose, raffinose, and stachyose, found in soy meal components. A highly active ${\alpha}$-galactosidase (${\alpha}$-d-galactoside galactohydrolase, EC 3.2.1.22) was isolated from a bacterium in a traditional Korean fermented medicinal herb preparation. The isolate, T2-16, was identified as Bacillus coagulans based on its 16S rRNA sequence and biochemical properties, and the strain was named Bacillus coagulans NRR-1207. When incubated in 10%(w/v) skim milk, Bacillus coagulans NRR1207 caused a decrease in the pH of the culture medium, as well as an increase in titratable acidity and viable cell counts. This strain also showed higher activities of ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\alpha}$-glucosidase, naphthol-AS-BO-phosphohydrolase, and acid phosphatase when compared to other enzymes. It hydrolyzed oligomeric substrates, such as raffinose and stachyose, and liberated galactose, indicating that the Bacillus coagulans NRR1207 ${\alpha}$-galactosidase hydrolyzed the ${\alpha}$-1,6 glycoside linkage. These results suggest that the decreased stachyose and raffinose contents observed in fermented soy meal are due to this ${\alpha}$-galactosidase activity. Bacillus coagulans NRR1207 therefore has potential probiotic activity and could be utilized in feed manufacturing, as well as for hydrolyzing non-digestible soy meal components.

• Journal of Food Hygiene and Safety
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• v.34 no.4
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• pp.367-373
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• 2019

In order to evaluate the microbiological safety of ice cream products, the total viable bacterial counts were measured in 6 kinds of ice pops, 5 kinds of non-milk fat ice cream, and 5 kinds of milk fat ice cream, sold in local markets. In addition, E. coli, S. aureus, B. cereus, and L. monocytogenes were artificially inoculated in three types of ice cream products and stored at $-5^{\circ}C$, $-10^{\circ}C$, and $-18^{\circ}C$, respectively, and after inoculation, viable cells were measured periodically. As a result of the total viable count, about 1~2 log CFU/mL was detected in 16 kinds of ice cream products. As a result of inoculation with microorganisms at various temperatures, the number of viable cells decreased as the storage period became longer, and the higher the storage temperature, the faster the microorganisms died. Especially, the microorganisms were killed faster in the ice pop products than in the other ice cream products, and the microorganisms were killed relatively slower in the milk ice cream. L. monocytogenes and S. aureus were relatively stable in frozen conditions compared to other microorganisms. The microbial contamination of commercial ice cream was lower than the allowable standard of the Korean Food Code. Microorganisms did not proliferate when the microorganism was inoculated at freezing temperature. Therefore, it is expected that the microbiological safety of frozen foods will be ensured if the sanitary control and disinfection of raw materials are thoroughly carried out during the production of frozen confections and the temperature control during distribution and storage is well maintained.

• Journal of Plant Biotechnology
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• v.46 no.3
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• pp.236-245
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• 2019

Fruit ripening is a genetically programmed process involving a number of biochemical and physiological processes assisted by variations in gene expression and enzyme activities. This process generally affects the phytochemical profile and the bioactive principles in fruits and vegetables. To appraise the variation in bioactive principles of fruits from Rosa rugosa during its ripening process, we analyzed the changes in antioxidant and anti-elastase activities and polyphenolic compounds during the four ripening stages of fruits. Overall, an extract of unripe fruits contained the highest levels of total phenolic and flavonoid contents, radical scavenging activity, reducing power, oxygen radical antioxidant capacity, and elastase inhibitory activity, compared with the extracts of fruits at other stages of ripening. Additionally, we found that the reduction of flavonoid content occurs because of decreased transcriptional levels of genes involved in flavonoid biosynthesis pathway during the ripening process. Based on HPLC analysis, we found that the extract of unripe fruits contained the highest amount of myricetin, caffeic acid, chlorogenic acid, syringic acid, and p-coumaric acid and suggested that the antioxidant and anti-elastase activities of the extract obtained from stage 1, should be mediated by the presence of these compounds. Additionally, we analyzed the interaction sites and patterns between these compounds and elastase using the structure-based molecular docking approach, and suggested that chlorogenic acid strongly interacted with elastase. Together, these findings suggest that the maturity of fruits has profound effects on the pharmaceutical value of R. rugosa.