• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.33-45
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• 2011

Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of cancer cell. Inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the cancer cell was reported. However, its role during oocyte maturation and early embryo development is very insufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on meiotic maturation and early embryonic development in pigs. We also investigated several indicators of developmental potential, including structural integrity, gene expression (Hsp90-, cell cycle-, and apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Then, we examined the roles of Hsp90 inhibitor on viability of primary cells in pigs. Porcine oocytes were cultured in the NCSU-23 medium with or without 17-AAG for 44 h. The proportion of GV arrested oocytes was significantly different between the 17-AAG treated and untreated group (78.2 vs 34.8%, p<0.05). After completion of meiotic maturation, the proportion of MII oocytes was lower in the 17-AAG treated group than in the control group (27.9 vs 71.0%, p<0.05). After IVF, the percentage of penetrated oocytes was significantly lower in the 17-AAG treated group (25.2%), resulting in lower normal pronucleus formation (2PN of 14.6%). Therefore, the inhibition of meiotic progression by Hsp90 inhibitor played a critical role in fertilization status. Porcine embryo were cultured in the PZM-3 medium with or without 17-AAG for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (7.5 vs 4.4, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. The mRNA expressions of cell cycle-related genes were down-regulated in the 17-AAG treated group compared with control. Also, the expression of the pro-apoptotic gene Bax increased in 17-AAG treated group, whereas expression of the anti-apoptotic gene Bel-XL decreased. However, the expression of ER stress-related genes did not changed by 17-AAG. Cultured pESF cells were treated with or without 17-AAG and used for MIT assay. The results showed that viability of pESF cells were decreased by treatment of 17-AAG ($2{\mu}M$) for 24 hr. These results indicated that 17-AAG decreased cell proliferation and increased cell death. Expression patterns Hsp90 complex genes (Hsp70 and p23), cell cycle-related genes (cdc2 and cdc25c) and apoptosis-related genes (Bax and Bcl-XL) were significantly changed by using RT-PCR analysis. The spliced form of pXbp-1 product (pXbp-1s) was detected in the tunicamycin (TM) treated cells, but it is not detected in 17-AAG treated cells. In conclusion, Hsp90 appears to play a direct role in porcine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with cell cycle- and apoptosis-related genes expression in developing porcine embryos.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.4
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• pp.575-580
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• 1996

The purpose of this study was to investigate the effect of dietary vitamin E levels on the enzymes such as superoxide dismutase(SOD), glutathione peroxidase(GSH-Px) and glutathione S-transferase (G57) involved in antioxidative defense system and lipid peroxidation in brain of cadmium administered rats. Sprague-Dawely male rats weighing $60\pm5g$ were divided into control and experimental groups. The experimental groups were divided into Cd-0E(vitamin E free diet), Cd-40E(40mg vitamin E/kg diet) and Cd-400E(400mg vitamin E/kg diet) according to the level of vitamin E supplementation. After each group was fed diet ad libitum for 2 or 4 weeks, 2.5mg cadmium per kg body weight was injected intraperitoneally once a day for 4 days. The rats were sacrificed for examination on the next day after the last injection of cadmium. The results are as follows: SOD activities of rat brain were lower in Cd-0E, but had a similar tendency to Cd-40E, Cd-400E groups compared with control group. GSH-Px acivities of rat brain were decreased in Cd-400E, Cd-40E and Cd-0E groups. GST activities of rat brain were decreased in Cd-0E, Cd-40E groups and not significantly different in Cd-400E compared with control group. Thiobarbituric acid reactive substances(TBARS) of rat brain was increased in Cd-0E, Cd-40E, Cd-400E in that order, TBARS was lower in Cd-40E, and Cd-400E by 28.8% and 44%, respectively, than Cd-0E group. The present result suggests that high level of dietary vitamin E protects against lipid peroxidative damage in rat induced by cadmium.

• Journal of Environmental Health Sciences
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• v.37 no.6
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• pp.429-439
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• 2011

Objectives: Ethylene oxide (EtO) is classified as a human carcinogen, but EtO is still widely used to sterilize heat-sensitive materials in hospitals. Employees working around sterilizers are exposed to EtO after sterilization. The aim of the present study was to assess the exposure of EtO level, coupled with occupationally induced micronuclei from hospital workers. The influence of genetic polymorphisms of detoxifying genes (GSTT1 and GSTM1) and DNA repair genes (XRCC1 and XRCC3) on the frequencies of micronuclei in relation to exposure of EtO was also investigated. Methods: The study population was composed of 35 occupationally exposed workers to EtO, 18 student controls and 44 unexposed hospital controls in Korea. Exposure to EtO is measured by passive personal samplers. We analyzed the frequencies of micronuclei by performing cytokinesis-block micronucleus assay (CBMN assay) and GSTM1, GSTT1, XRCC1, and XRCC3 were also genotyped by performing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: The frequencies of micronuclei in EtO exposure group, student controls and hospital controls were $18.00{\pm}7.73$, $10.47{\pm}7.96$ and $13.86{\pm}6.35$ respectively and their differences were statistically significant, but no significant differences according to the level of EtO were observed. There was a dose-response relationship between the frequencies of micronuclei and cumulative dose of EtO, but no significantly differences were observed. We also investigated the influence of genetic polymorphisms (GSTM1, GSTT1, XRCC1, and XRCC3) on the frequencies of micronuclei, but there were no differences in the frequencies of micronuclei by genetic polymorphisms. Conclusions: The frequencies of micronuclei in EtO exposure group was significantly higher than control groups. A dose-response relationship was found between the level of EtO exposure and the frequencies of micronuclei, but no statistically differences were observed. We also found that the frequencies of micronuclei were increased according to cumulative EtO level. There was no association of the genetic GSTM1, GSTT1, XRCC1, and XRCC3 state with the frequency of micronuclei induced by EtO exposure.

• Korean Journal of Poultry Science
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• v.37 no.1
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• pp.15-21
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• 2010

To investigate the effect of dietary supplementation of Acanthopanax senticosus (AS) and Eucommia ulmoides (EU) on antioxidant defense system in laying hens, a total of three hundreds sixty 20-wk old Hyline brown commercial laying hens were assigned to five dietary groups for 10-wk: (1) control diet, (2) control diet supplemented with AS at 0.5%, (3) control diet supplemented with AS at 1.0%, (4) control diet supplemented with EU at 0.5% and (5) control diet supplemented with EU at 1.0%. Total antioxidant status (TAS) in blood and antioxidant enzymes including superoxide dismutase (SOD), gluthathione -S- transferase (GST) and glutathione peroxidase (GSH-Px), and lipid peroxidation in the small intestine and liver were measured. There were no changes in body weight for 10-wk dietary treatment. TAS in blood significantly (P<0.05) increased in birds fed the diet supplemented with 1% AS and 0.5 and 1.0% EU compared with those fed control diet. Especially, dietary EU showed much higher (P<0.05) TAS compared with AS. In the antioxidant defense enzymes, GST activity of the small intestine was shown to be significantly (P<0.05) increased in birds fed the diets supplemented with 0.5 and 1.0% EU compared with those fed the control diet. In addition, intestinal SOD activity significantly (P<0.05) increased in birds fed the diets supplemented with 0.5% of AS and EU. However, we could not observe any significant dietary treatment effect of those antioxidant parameters in the liver. In conclusion, dietary supplementation of 0.5% AS and EU in a laying hen diet could be applied as a potential antioxidant source to improves bio-activity of antioxidant and economical aspect in laying hens.

• Korean Journal of Poultry Science
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• v.37 no.1
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• pp.23-33
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• 2010

The current study was performed to develop natural bio-active substances as additives for the production of high quality broiler chickens. A total of 120 male 3 day-old broiler chicks were randomly allocated to CON (control), GK2.5 (ginkgo leaf 2.5%), GK5.0 (ginkgo leaf 5.0%), PK2.5 (pumpkin 2.5%) and PK5.0 (pumpkin 5.0%) of five groups in cages (24 birds per group). All birds were fed corresponding diets from 3 to 35 d of age and determined growth performance and biological parameters including blood biochemical profiles, antioxidant status and intestinal microflora. During the entire feeding trial, GK5.0 and PK5.0 groups resulted in a significantly (P<0.05) higher FCR than GK2.5 and PK2.5 groups. Plasma triglyceride significantly (P<0.05) increased in GK5.0 group compared with the other groups, and the level of alanine transaminase (ALT) increased (P<0.05) in GK5.0 and PK5.0 groups compared with that in PK2.5 group. Dietary addition of ginkgo leaf and pumpkin significantly (P<0.05) increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the small intestine. Also, the addition of 2.5% ginkgo leaf significantly (P<0.05) increased the activities of SOD, GSH-Px and glutathione-S-transferase (GST) in the liver. Futhermore, muscle GST activity significantly (P<0.05) enhanced by dietary addition of ginko leaf and pumpkin. However, the level of lipid peroxidation (MDA) in the small intestine and muscle turned to be higher (P<0.05) in PK5.0 group. The colony forming units (CFU) of E. coli in intestinal digesta significantly (P<0.05) decreased in both ginko leaf and pumpkin supplemented groups compared with CON group. In conclusion, dietary addition of 2.5% ginko leaf and pumpkin as dietary sources can be applicable for the production of high quality broiler chickens.

• Journal of Nutrition and Health
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• v.36 no.8
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• pp.801-810
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• 2003

Chronic alcoholism is considered a common cause of malnutrition. Especially, micronutrient deficiency may playa critical role in the incidence of alcoholic liver diseases. This study was conducted to investigate the effect of folate deficiency and ethanol consumption on cholesterol metabolism and the antioxidative system in rats. Plasma concentration of total cholesterol was increased by ethanol administration in folate-fed rats. HDL-cholesterol tended to be higher in the folate-fed group, but it was not significant. The plasma and hepatic levels of malondialdehyde were increased after chronic ethanol feeding, but dietary folate depressed the plasma malondialdehyde content of rats. Ethanol or folate feeding did not significantly change alcohol dehydrogenase activity. But folate feeding increased catalase activity in ethanol-fed rats. There was no significant change in superoxide dismutase activity among the experimental groups. Glutathione peroxidase activity tended to decrease by chronic ethanol feeding, but dietary folate did not affectthe glutathione peroxidase activity of chronic ethanol-fed rats. Glutathionine-S-transferase activity was not affected by ethanol feeding or folate deficiency. The plasma and hepatic levels of retinol decreased after chronic ethanol feeding. The hepatic level of retinol significantly decreased in ethanol-fed rats by folate deficiency. The plasma level of $\alpha$-tocopherol tended to be low in the folate deficient group with ethanol feeding, but there was no difference among the experimental groups in the hepatic level of $\alpha$-tocopherol. These results demonstrate that chronic ethanol consumption changes the plasma cholesterol metabolism and antioxidative system of rats, and optimal folate feeding in ethanol-fed rats exerts protective effects to some extent.

• Journal of Nutrition and Health
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• v.40 no.7
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• pp.606-615
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• 2007

This study was performed to investigate anti-thrombogenic, anti-inflammatory effects of n-BuOH (B) and $CH_2Cl_2$ (MC) fractions extracted from Sancho (Zanthoxylum. schinifolium) leaves in rats fed high fat diets. The experimental animal groups were consisted of eight including one 5% fat (N) and one 20% fat (H) without the test materials in diets and six H groups of feeding three levels (50, 100 and 150 mg/day) of the B and the MC fractions from Z. schinifolium, respectively. Plasma activated partial thromboplastin times and thrombin times of H group were decreased compared to the N group, but they were increased by feeding the MC fraction of 50 mg and over. Polymorphonuclear leukocyte 5#-lipo-xygenase activities and leukotriene $B_4$ contents of the H group were significantly increased compared to the N group, but they were decreased in the 100 mg and 150 mg of B fraction or the 150 mg of MC fraction fed groups. Liver cytochrome $P_{450}$, $O_2^-$, $H_2O_2$ and GSSG contents were increased by the high fat diet but decreased by feeding the B fraction or the MC fraction, while GSH content and glutathione S-transferase activity lowered by high fat diet were increased by feeding the two solvent fractions. The effects of the solvent fractions were evident at the level of 100 mg/day and over. The present results confirmed that two solvent fractions from the leaves of Z, schinifolium have enhancing effects on anti-thrombosis and anti-inflammation partly by antioxidant action and partly by direct modulation of the respective processeds. In conclusion, the n-BuOH and $CH_2Cl_2$ fractions from leaves of Z, schinifolium can be utilized as the proper ingredients of functional foods for preventing chronic degenerative disease.

• Journal of the Korean Academy of Child and Adolescent Psychiatry
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• v.15 no.2
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• pp.178-184
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• 2004

Objectives : This study was conducted to investigate the association of the COMT polymorphism with the TD in Korean sample of families with TD probands. The relationship between risk alleles and specific clinical features (tic severity, comorbidity, drug response) was also explored. Method : Patients were recruited from the Tic Disorder clinic at the Child & Adolescent Psychiatric Division of Seoul National University Hospital and assessed through 2 stage evaluation. Firstly, all the patients and parents received semistructured interview using Korean version of K-SADS-PL. Secondly all the patients received clinical interview and tic severity assessment with Korean version of YGTSS. The subjects in control group were recruited from the health promotion center in out hospital and were evaluated by SCL-90 and SCID-IV. Through these process, total of 42 children and adolescents with TD, their 84 parents and 86 control subjects were finally recruited. Genotyping for The Val158Met polymorphism of the COMT gene was done by standardized method. After collection of genetic data of all the patients, parents and control subjects, case-control comparison and tranmission dysequilibrium test was executed by SPSS version 11. Result : From the case-control comparison, the frequency of L-allele and LL genotype was significantly higher in TD group. However, no differences were found from the TDT. No significant differences were found in in family history of tic, ADHD, OCD, drug response and comorbid conditions among the three different genotypes in patients with TD. Conclusion : Though this study results should be interpreted cautiously due to small sample size and negative finding in TDT test, this study is the first report that there is positive association between the functional polymorphism of COMT gene the TD.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.7
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• pp.1043-1053
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• 2013

We investigated the ability of soybean curd residue (SCR) and its fermented products to inhibit obesity and improve the blood lipid profiles of obese mice fed a high-fat diet. Samples were prepared by fermenting SCR with Aspergillus oryzae var effuses KACC 44990 (ASCR), a microbe used for the fermentation of traditional Korean Meju, and with Monascus pilosus IFO 4480 (MSCR), a microbe used for the production of red rice. In addition, AMSCR, a mixture composed of equal amounts of ASCR and MSCR, was also prepared. Male mice were divided into six groups and fed with either a normal diet, a high-fat diet, or a high-fat diet supplemented with SCR, ASCR, MSCR, or AMSCR. After 8 weeks, body weight gain, serum and hepatic lipid profiles, and the activities of enzymes that generate or scavenge reactive oxygen species (ROS) were evaluated. Compared with the high-fat diet group, all the test groups showed a significant reduction in body, organ, and epididymal fat weight gain. These effects were observed with supplements in the order AMSCR>ASCR>MSCR>SCR. Similarly, supplements of test samples reduced high levels of serum and hepatic triglycerides (TG), total cholesterol, and low-density lipoprotein (LDL) cholesterol caused by hight-fat diet, while high-density lipoprotein (HDL) cholesterol was increased. Interestingly, the ability of ASCR to lower serum TG was stronger than that of MSCR, while MSCR showed a stronger hypocholesterolemic effect than ASCR. Meanwhile, AMSCR returned comprehensively serum lipid levels to normal. In addition, hepatic damage was prevented with effects in the order AMSCR>ASCR>MSCR>SCR. Hepatic ROS generating system including xanthine oxidase (XO) and ROS scavenging system including superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione S-transferase (GST) were recovered to normal level by all test diets. In conclusion, this study suggests that SCR and its fermented products can inhibit obesity and improve lipid profiles.

• Tuberculosis and Respiratory Diseases
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• v.55 no.1
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• pp.88-97
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• 2003

Background : Although smoking is a major cause of chronic obstructive pulmonary disease (COPD), only 10-20% of cigarette smokers develop symptomatic COPD, which suggests the presence of genetic susceptibility. This genetic susceptibility to COPD might depend on variations in the activities of the enzyme that detoxify hazardous chemical products, such as microsomal epoxide hydrolase (mEPHX) and glutathione-S transferase M1 subunit (GSTM1) genes. Methods : The genotypes of 58 patients with COPD, and 79 age matched control subjects, were determined by a polymerase chain reaction, followed by restriction fragment length polymorphism (PCR-RFLP) for the mEPHX, and multiplex PCR for the GSTM1. Results : GSTM1 was deleted in 53.3% of the subjects. There was no difference in GSTM1 deletion rates between the COPD patients (32/58, 55.2%) and the control subjects (41/79, 51.9%). The combination patterns of two polymorphisms of mEPHX showed slow enzyme activity in 29(21.2%), normal in 73(53.3%) and fast in 32(23.4%). The COPD group (7/57, 12.3%) showed a significantly lower incidence of slow enzyme activity compared to the control subjects (22/77, 28.6%, p<0.05). However, when the COPD and control groups were compared with smokers only, there were no significant differences in the genotypes of GSTM1 and mEPHX. Conclusion : The genotypes of GSTM1 and mEPHX were not significant risk factors of COPD in this cohort of study.