• Journal of the East Asian Society of Dietary Life
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• v.21 no.5
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• pp.731-738
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• 2011

Cholesterol is the precursor of various steroid hormones, bile acid, and vitamin D with functions related to regulation of membrane permeability and fluidity. However, the presence of excess blood cholesterol may lead to arteriosclerosis and hypertension. Moreover, dietary cholesterol may affect blood cholesterol levels. Generally, cholesterol determination is performed by spectrophotometric or chromatographic methods, but these methods are very time consuming and costly, and require complicated pretreatment. Thus, the development of a rapid and simple analysis method for measuring cholesterol concentration in food is needed. Multi-walled carbon nanotube (MWCNT) was functionalized to MWCNT-$NH_2$ via MWCNT-COOH to have high sensitivity to $H_2O_2$. The fabricated MWCNT-$NH_2$ was attached to a glassy carbon electrode (GCE), after which Prussian blue (PB) was coated onto MWCNT-$NH_2$/GCE. MWCNT-$NH_2$/PB/GCE was used as a working electrode. An Ag/AgCl electrode and Pt wire were used as a reference electrode and counter electrode, respectively. The sensitivity of the modified working electrode was determined based on the amount of current according to the concentration of $H_2O_2$. The response increased with an increase of $H_2O_2$ concentration in the range of 0.5~500 ${\mu}M$ ($r^2$=0.96) with a detection limit of 0.1 ${\mu}M$. Cholesterol oxidase was immobilized to aminopropyl glass beads, CNBr-activated sepharose, Na-alginate, and toyopearl beads. The immobilized enzyme reactors with aminopropyl glass beads and CNBr-activated sepharose showed linearity in the range of 1~100 ${\mu}M$ cholesterol. Na-alginate and toyopearl beads showed linearity in the range of 5~50 and 1~50 ${\mu}M$ cholesterol, respectively. The detection limit of all immobilized enzyme reactors was 1 ${\mu}M$. These enzyme reactors showed high sensitivity; especially, the enzyme reactors with CNBr-activated sepharose and Na-alginate indicated high coupling efficiency and sensitivity. Therefore, both of the enzyme reactors are more suitable for a cholesterol biosensor system.

• Journal of Food Hygiene and Safety
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• v.26 no.3
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• pp.273-282
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• 2011

This study established hazards which may cause risk to human at farm during cultivation stage of paprika. Samples of plants (paprika, leaf, stem), cultivation environments (water, soil), personal hygiene (hand, glove, clothes), work utensils (carpet, basket, box) and airborne bacteria were collected from three paprika farms (A, B, C) located in Western Gyeongnam, Korea. The collected samples were assessed for biological (sanitary indications and major foodborne pathogens), chemical (heavy metals, pesticide residues) and physical hazards. In biological hazards, total bacteria and coliform were detected at the levels of 1.9~6.6 and 0.0~4.610g CFU/g, leaf, mL, hand or 100 $cm^2$, while Escherichia coli was not detected in all samples. In major pathogens, only Bacillus cereus were detected at levels of ${\leq}$ 1.5 log CFU/g, mL, hand or 100 $cm^2$, while Staphylococuus aureus, Listeria monocytogenes, E. coli O157 and Salmonella spp. were not detected in all samples. Heavy metal and pesticide residue as chemical hazards were detected at levels below the regulation limit, physical hazard factors, such as insects, pieces of metal and glasses, were also found in paprika farms. Proper management is needed to prevent biological hazards due to cross-contamination while physical and chemical hazards were appropriate GAP criteria.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.9
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• pp.1257-1264
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• 2016

Extracts from Artemisia annua $Linn\acute{e}$ (AAE) have various functions (anti-malaria, anti-virus, and anti-oxidant). However, the mechanism of the effects of AAE is not well known. Thus, we determined the apoptotic effects of AAE in AGS human gastric carcinoma cells. In this study, we suggested that AAE may exert cancer cell apoptosis through the Akt/mammalian target of rapamycin (mTOR)/glycogen synthase kinase (GSK)-$3{\beta}$ signal pathway and mitochondria-mediated apoptotic proteins. Activation by Akt phosphorylation resulted in cell proliferation through phosphorylation of tuberous sclerosis complex 2 (TSC2), mTOR, and GSK-$3{\beta}$. Thus, de-phosphorylation of Akt inhibited cell proliferation and induced apoptosis through inhibition of Akt, mTOR, phosphorylation of GSK-$3{\beta}$ at serine9, and control of Bcl-2 family members. Inhibition of GSK-$3{\beta}$ attenuated loss of mitochondrial membrane potential and release of cytochrome C. Bax and pro-apoptotic proteins were activated by their translocation into mitochondria from the cytosol. Translocation of Bax induced outer membrane transmission and generated apoptosis through cytochrome C release and caspase activity. We also measured 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, lactate dehydrogenase assay, Hoechst 33342 staining, Annexin V-PI staining, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide staining, and Western blotting. Accordingly, our study showed that AAE treatment to AGS cells resulted in inhibition of Akt, TSC2, GSK-$3{\beta}$-phosphorylated, Bim, Bcl-2, and pro-caspase 3 as well as activation of Bax and Bak expression. These results indicate that AAE induced apoptosis via a mitochondrial event through regulation of the Akt/mTOR/GSK-$3{\beta}$ signaling pathways.

• Journal of Life Science
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• v.29 no.12
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• pp.1371-1377
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• 2019

Oxidative stress induced by the over-production of reactive oxygen species (ROS) in the brain is the most common cause of neurodegenerative diseases such as Alzheimer's. In the present study, we investigated the protective effects of flavonoids and their glycosides, namely kaempferol, kaempferol-3-O-glucoside, quercetin, and quercetin-3-β-D-glucoside, against H₂O₂-induced oxidative stress in the C6 glial cells. The H₂O₂-treated glial cells exhibited decreased cell viability and increased ROS production when compared with normal cells. However, cells treated with each of the four flavonoids/glycosides demonstrated significantly increased viability and suppressed ROS production when compared with the H₂O₂-treated control group. These results indicate that flavonoids/glycosides attenuate oxidative stress induced by H₂O₂ in C6 glial cells. To confirm the protective molecular mechanisms, we measured pro-inflammatory factors such as inducible nitric oxide synthase, cyclooxygenase-2, and interleukin-1β. H₂O₂ treatment was seen to elevate these factors and decrease IκB-α in the C6 glial cells, while the flavonoids/glycosides induced a down-regulation of the pro-inflammatory factors and increased IκB-α, indicating a neuroprotective effects through attenuation of the inflammation. In particular, quercetin and its glycoside showed a higher neuroprotective effect than the kaempferol treatments. These results suggest that these flavonoids and their glycosides could be promising therapeutic agents for neurodegenerative diseases via the attenuation of oxidative stress.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.5
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• pp.625-634
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• 2011

Green tea seed coat (GTSC) was extracted with 100% ethanol for 4 hr and then fractionated with petroleum ether (PE), ethyl acetate (EtOAC) and butanol (BuOH). The EtOAC fraction showed the highest level in total phenol contents and the lowest level in nitric oxide (NO) production in LPS-stimulated RAW264.7 macrophage cell. Thus, this study was carried out to investigate the anti-inflammatory and its mechanisms of GTSC EtOAC fraction in LPS-stimulated RAW264.7 macrophage cell. GTSC EtOAC fraction contained EGC ($1146.48{\pm}11.01\;{\mu}g/g$), tannic acid ($966.99{\pm}32.24\;{\mu}g/g$), EC ($70.88{\pm}4.39\;{\mu}g/g$), gallic acid ($947.61{\pm}1.03\;{\mu}g/g$), caffeic acid ($37.69{\pm}1.46\;{\mu}g/g$), ECG ($35.46{\pm}3.19\;{\mu}g/g$), and EGCG ($15.53{\pm}0.09\;{\mu}g/g$) when analyzed by HPLC. NO production was significantly (p<0.05) suppressed in a dose-dependent manner with an $IC_{50}$ of $80.11\;{\mu}g$/mL. Also prostaglandin $E_2$ level was also inhibited in a dose-dependent manner. Moreover, iNOS protein expression was suppressed in dose-dependent manner but COX-2 gene expression was not affected. Total antioxidant capacity and glutathione (GSH) levels were enhanced more than the LPS-control. Expressions of antioxidative enzymes including catalase, GSH-reductase and Mn-SOD were elevated compared to LPS-control. Nuclear p65 level was decreased in the GTSC EtOAC fraction in a dose-dependent manner. These results indicate that GTSC EtOAC fraction inhibit oxidative stress and inflammatory responses through elevated GSH levels, antioxidative enzymes expressions and suppression of iNOS expression via NF-${\kappa}B$ down-regulation.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.4
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• pp.417-425
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• 2017

Cnidium monnieri (L.) Cusson is distributed in China and Korea, and the fruit of C. monnieri is used as traditional Chinese medicine to treat carbuncle and pain in female genitalia. In this study, we examined the anti-proliferation and cell cycle arrest effects of ethanol extracts from C. monnieri (CME) in AGS gastric cancer cells. Our results show that CME suppressed cell proliferation and induced release of lactate dehydrogenase (LDH) in AGS cells by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay and LDH assay. Cell morphology was altered by CME in a dose-dependent manner. In order to identify the cell cycle arrest effects of CME, we investigated cell cycle analysis after CME treatment. In our results, CME induced cell cycle arrest at G1 phase. Protein kinase B (Akt) plays a major role in cell survival mechanisms such as growth, division, and metastasis. Akt protein regulates various downstream proteins such as glycogen synthase kinase-$3{\beta}$ (GSK-$3{\beta}$) and tumor protein p53 (p53). Expression levels of p-Akt, p-GSK-$3{\beta}$, p53, p21, cyclin E, and cyclin-dependent kinase 2 (CDK2) were determined by Western blot analysis. Protein levels of p-Akt, p-GSK-$3{\beta}$, and cyclin E were reduced while those of p53, p21, and p-CDK2 (T14/Y15) were elevated by CME. Moreover, treatment with CME, LY294002 (phosphoinositide 3-kinase/Akt inhibitor), BIO (GSK-$3{\beta}$ inhibitor), and Pifithrin-${\alpha}$ (p53 inhibitor) showed that cell cycle arrest effects were mediated through regulation of the Akt/GSK-$3{\beta}$/p53 signaling pathway. These results suggest that CME induces cell cycle arrest at G1 phase via the Akt/GSK-$3{\beta}$/p53 signaling pathway in AGS gastric cancer cells.

• Journal of Life Science
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• v.17 no.12
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• pp.1689-1694
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• 2007

This study was conducted to investigate both fat-soluble and water-soluble vitamin contents of commercial powdered infant formula for obtaining basic data on infant nutrition. Ten commercial infant formula based on cow's milk were collected and the contents of fat-soluble vitamins (vitamin A, D, E, K) and water-soluble vitamins (vitamin C, thiamin, riboflavin, niacin, $B_6$, folate, $B_{12}$, pantothenic acid, biotin) were compared with Dietary Reference Intakes for Koreans (KDRIs). The overall vitamin contents in 100 g and in 100 kcal of infant formula satisfied the recommended formula regulation (KDRIs) and Codex. In infant formula during 0-5 monthly age, fat-soluble vitamin A, D, E, K could supply 178.6%, 205.3%, 208.4%, 976.3% of adequate daily vitamin intakes, respectively. Water soluble vitamins, vitamin C, thiamin, riboflavin, niacin, $B_6$, folate, $B_{12}$, pantothenic acid, biotin could supply 173.2%, 237.2%, 269.8%, 295.9%, 431.6%, 165.8%, 1186.3%, 203.8%, 408.3% of adequate daily vitamin intakes, respectively. In infant formula during 6-11 monthly age, all vitamins satisfied their adequate daily intakes as well. Vitamin A, D, E, K supplied 199.2%, 262.3%, 220.5%, 626.46% of adequate daily vitamin intakes. Vitamin C, thiamin, riboflavin, niacin, $B_6$, folate, $B_{12}$, pantothenic acid, biotin could supply 179.5%, 210.2%, 264.7%, 241.5%, 206.0%, 166.9%, 699.5%, 247.0%, 475.0% of adequate intake of KDRIs. From this study, evaluation of vitamin contents of commercial infant formula was established, which could strengthen the basic information on infant nutrition.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.8
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• pp.303-312
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• 2019

Oxidative stress in kidneys can precede the development of chronic renal injury. We investigated the antioxidative effect of membrane-free stem cell extract (MFSCE) from adipose tissue in LLC-$PK_1$ renal proximal tubule cells. Treatment of LLC-$PK_1$ cells with MFSCE showed the up-regulation of heme-oxygenase-1, thioredoxin reductase 1, and NADPH quinine oxidoreductase-1 protein expressions, which are proteins related with antioxidative activities. When oxidative stress was induced by 3-morpholinosydnonimine (SIN-1), cell viability was decreased, indicating that LLC-$PK_1$ cells were damaged by SIN-1. However, MFSCE significantly elevated cell viability from 58.84% to 64.43% at the concentration of $2.5{\mu}g/mL$ in oxidative stress-induced LLC-$PK_1$ cells. Furthermore, MFSCE ameliorated inflammation and apoptosis in SIN-1-treated LLC-$PK_1$ cells by modulating protein expressions. Inducible nitric oxide synthase and cyclooxygenase-2 protein expressions were down-regulated when LLC-$PK_1$ cells were treated with MFSCE. Apoptosis-related proteins, including B-cell lymphoma-2-associated X protein/B-cell lymphoma-2 ratio, cleaved caspase-3, and cleaved-poly (ADP-ribose) polymerase, were also down-regulated. It indicated that MFSCE protected apoptosis against oxidative stress in LLC-$PK_1$ cells. Taken together, these results suggested that MFSCE had a protective effect against SIN-1-induced oxidative stress in LLC-$PK_1$ cells. Therefore, MFSCE could be a promising therapeutic agent for oxidative stress-induced renal injury.

• Korean Journal of Environmental Agriculture
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• v.25 no.1
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• pp.77-84
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• 2006

This study was conducted to find a system for screening organic resources with 16 species, 62 samples which were selected to randomizing point from city, province and industrial areas in the whole country. Content of organic matters were $65.3%{\sim}98.0%$ in all samples so that they were largely over than 60%, raw material regulation of compost. Concentrations of total nitrogen and total phosphorus were $0.7{\sim}4.8%\;and\;0.8{\sim}5.0$, they could look for effect of the nitrogen and phosphorus supply as a raw material of compost. In case of 8 elements concentrations of heavy metal, they were too high to use as raw materials of compost which were over to regulation limit in Cu, Cr, Ni, and As from fiber industry, Ni from food company and leather industry, and the others are adapt to limit levels. HEM contents fro the highest to $113mg\;kg^{-1}$ from liber industry and PAHs content were the highest to $3,462ug\;kg^{-1}$ from paper-mill manufacture. Distribution of PAHs concentiations were naphthalene>phenanthrene>pyrene>fluoroanthene>acenaphthene. $Microtox{(R)}\;EC_{50}$ values for bioassay were pharmaceutical company>paper-mill manufacture>industrial area sewage sludge>fiber industry>urban sewage sludge>metropolitan sewage sludge. HEM between Zn, Cu, and Ni was significant at the 99% and between Cd was significant at the 95%, Microtox between Hg and BEM significant at the 95%.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.5
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• pp.747-754
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• 2005

Nutritional regulation of gene expression associated with growth and feeding behavior in avian species can become an important technique to improve poultry production according to the supply of nutrients in the diet. Insulin-like growth factor-I (IGF-I) found in chickens has been characterized to be a 70 amino acid polypeptide and plays an important role in growth and metabolism. Although it is been well known that IGF-I is highly associated with embryonic development and post-hatching growth, changes in the distribution of IGF-I gene expression throughout early- to late-embryogenesis have not been studied so far. We revealed that the developmental pattern of IGF-I gene expression during embryogenesis differed among various tissues. No bands of IGF-I mRNA were detected in embryonic liver at 7 days of incubation, and thereafter the amount of hepatic IGF-I mRNA was increased from 14 to 20 days of incubation. In eyes, a peak in IGF-I mRNA levels occurred at mid-embryogenesis, but by contrast, IGF-I mRNA was barely detectable in the heart throughout all incubation periods. In the muscle, no significant difference in IGF-I gene expression was observed during different stages of embryogenesis. After hatching, hepatic IGF-I gene expression as well as plasma IGF-I concentration increases rapidly with age, reaches a peak before sexual maturity, and then declines. The IGF-I gene expression is very sensitive to changes in nutritional conditions. Food-restriction and fasting decreased hepatic IGF-I gene expression and refeeding restored IGF-I gene expression to the level of fed chickens. Dietary protein is also a very strong factor in changing hepatic IGF-I gene expression. Refeeding with dietary protein alone successfully restored hepatic IGF-I gene expression of fasted chickens to the level of fed controls. In most circumstances, IGF-I makes a complex with specific high-affinity IGF-binding proteins (IGFBPs). So far, four different IGFBPs have been identified in avian species and the major IGFBP in chicken plasma has been reported to be IGFBP-2. We studied the relationship between nutritional status and IGFBP-2 gene expression in various tissues of young chickens. In the liver of fed chickens, almost no IGFBP-2 mRNA was detected. However, fasting markedly increased hepatic IGFBP-2 gene expression, and the level was reduced after refeeding. In the gizzard of well-fed young chickens, IGFBP-2 gene expression was detected and fasting significantly elevated gizzard IGFBP-2 mRNA levels to about double that of fed controls. After refeeding, gizzard IGFBP-2 gene expression decreased similar to hepatic IGFBP-2 gene expression. In the brain, IGFBP-2 mRNA was observed in fed chickens and had significantly decreased by fasting. In the kidney, IGFBP-2 gene expression was observed but not influenced by fasting and refeeding. Recently, we have demonstrated in vivo that gizzard and hepatic IGFBP-2 gene expression in fasted chickens was rapidly reduced by intravenous administration of insulin, as indicated that in young chickens the reduction in gizzard and hepatic IGFBP-2 gene expression in vivo stimulated by malnutrition may be, in part, regulated by means of the increase in plasma insulin concentration via an insulin-response element. The influence of dietary protein source (isolated soybean protein vs. casein) and the supplementation of essential amino acids on gizzard IGFBP-2 gene expression was examined. In both soybean protein and casein diet groups, the deficiency of essential amino acids stimulated chickens to increase gizzard IGFBP-2 gene expression. Although amino acid supplementation of a soybean protein diet significantly decreased gizzard IGFBP-2 mRNA levels, a similar reduction was not observed in chickens fed a casein diet supplemented with amino acids. This overview of nutritional regulation of IGF-I and IGFBP-2 gene expression in young chickens would serve for the establishment of the supply of nutrients to diets to improve poultry production.