• Korean Journal of Animal Reproduction
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• v.23 no.1
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• pp.75-84
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• 1999

This study assessed development in vitro of pronuclear(PN) stage embryos cryopreserved by the method of either vitrification or slow freezing, by using of different cryoprotectants, and equilibration and cooling rate, in rabbit. Ethyleneglycol- ficoll- sucrose(EFS) or ethyleneglycol- polyvinylpyrrolidone - galactose- (EPG-I) for vitrification, and EPG- II for slow freezing as cryoprotectant were used. The pronuclear embryos were exposed to EFS for 0 to 5 min and diluted with D-PBS and/or pre-dilution with 0.5 M sucrose. To examine the viability of frozen-thawed embryos, PN embryos were co-cultured with bovine oviductal epitherial cell(BOEC) for 5 days to hatching blastocyst stage in 39 $^{\circ}C$ 5% $CO_2$incubator. The results obtained were as follows: The dilution with 0.5 M sucrose and D-PBS after the exposure to EFS for 1.0 min resulted in no significant(P＜0.05) decrease in the development of PN embryos to hatching blastocyst(72.0%), compared with controls. The development of PN embryos cryopreserved to hatching blastocyst was not significantly (P＜0.05) different between EFS for 1.0 min(72.0%), EPG-I for 1.0 min(72.0%) and EPG-II for 30 min(66. 7%). The post-thaw development of PN embryos to hatching blastocyst was similarly very low as 6.1% and 11.5% in vitrification with EFS and slow freezing with EPG-II, respectively. The incidence of post-thaw zona-crack in PN embryos cryopreserved by slow freezing with plunging to liquid nitrogen at －35$^{\circ}C$ was signicantly(P＜0.05) higher(25.0%), compared with －85$^{\circ}C$ (1.9%). These results indicated that the rabbit PN embryos could be cryopreserved with either vitrification or slow freezing procedure, and frozen PN embryos could be successfully developed in vitro to haching blastocyst. but the post-thaw development of cryopreserved PN embryos was still very low under the present conditions.

• Journal of Society of Preventive Korean Medicine
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• v.9 no.2
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• pp.59-75
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• 2005

The experiments were undertaken to evaluate the effects of Bosaengtang in pregnant rats and fetuses. Female Sprague-Dawley rats were orally administered with Bosaengtang at the dose of 5mg/kg/day for 20 days. Pregnant rats were sacrificed at the 20th day of gestation, and observed internal and reproductive organs. Fetuses were randomly selected and fixed in 95% ethanol. Fetuses were stained with alcian blue and alizarin red S, and observed skeletal malformations. The results obtained were as follows : Bosaengtang administered group showed higher maternal body weight than the control group, but both groups showed increase in weight. Bosaengtang administered group showed lower than the control group, and higher liver and kidney weight than the control group, but the differences were minimal. There were no significant changes between the control and treated group in blood chemistry values and hematological values but all the groups were within in normal ranges. There were no significant changes in the number of corpus luteum, implantation, live fetus and implantation rate, delivery rate, late resorption rate, sex ratio, but Bosaengtang administered group showed higher early resorption rate than control group. comparing the control and Bosaengtang group, neonatal body weight and the number of fetuses were increased in Bosaengtang group. The fetuses of dams treated with Oriental medicine didn't showed external malformation. Vertebral and sternal variations were observed in Bosaengtang group, but the differences were not apparent compared to the control group. The number of ribs, cervical, thoracic and lumbar vertebrae were normal. The number of sacral was similar and the number of caudal was increased. Fetuses showed significant difference in the number of caudal vertebrae. (P<0.01) From these results, we can carefully conclude that Bosaengtang showed beneficial effects on maternal body weight, early resorption rate, number of live fetus. There were no significant changes in organ weight, hematoscopy, reproduction organs. External malformation wasn't visible. Skeletal variations were showed in vertebrae and sternum but compared to the control group, these variations weren't much different.

• Korean Journal of Microbiology
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• v.43 no.3
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• pp.151-158
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• 2007

The possibility of inadvertent introduction of therapeutic gene expressing viral vectors has raised safety concerns about germ-line infection. Particularly, for indications such as prostate cancer and ovarian cancer, the proximity of the point of viral administration to organs of the reproductive system raises concerns regarding inadvertent germ-line transmission of genes carried by the virus vector. To evaluate the safety of in vivo adenovirus mediated gene transfer, we explored the biodistribution, persistance and potential germ-line transmission of p53-expressing adenovirus (Ad-CMV-p53). Both male and female Balb/c mice were injected with $1{\times}10^9$ PFU of Ad-CMV-p53. The PCR analysis showed that there were detectable vector sequences in liver, kidney, spleen, seminal vesicle, epididymis, prostate, ovary, and uterus. The RT-PCR analysis for detecting inserted gene, p53 showed that Ad-CMV-p53 viral RNA were present in spleen, prostate and ovary. Direct injected male and female mice of adenovirus vector into testis and ovary were mated and their of offspring were evaluated for germ-line transmission of the adenoviral vector. The PCR and RT-PCR analysis showed no evidence of germline transmission, although vector sequences were detected in DNA extracted from gonadal tissues. Real-time PCR result confirmed a significant decrease of adenovirus in gonad tissues 1 week after injection. We have also analysed the cell specific localization of viral DNA in gonad tissues by using in-situ PCR. Positive signals were detected in interstitial tissue but not in seminiferous tubule in sperm. In the case of ovary, adenovirus signal were localized to the stromal tissue, but no follicular signals were observed. Together, these data provide strong evidence that the risk of the Inadvertent germ-line transmission of vector sequences following intraperitoneal or direct injection into genito-urinary system of adenovirus is extremely low.

• Journal of Food Hygiene and Safety
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• v.27 no.1
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• pp.96-102
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• 2012

This study was designed to evaluate a repeated oral dose toxicity and immunomodulating activity of $Pulsatilla$ $koreana$ and $Artemisiae$ $annuae$ in Sprague-Dawley rats. The female rats were treated with $Pulsatilla$ $koreana$ and $Artemisiae$ $annuae$ of control group, low group (0.5 ml/kg), medium group (1 ml/kg), high group (2 ml/kg) for distilled water, intragastrically for 4 weeks, respectively. To ensure the safety of $Pulsatilla$ $koreana $ and $Artemisiae$ $annuae$ such as the following were observed and tested. We examined the body weight, the feed intake, the clinical signs, the ophthalmological test, the hematological and the serum biochemical analysis. We also observed the histopathological changes of liver and kidney in rats. Hematological results were the increase of neutrophils, lymphocytes and monocytes in the high dose group of $Pulsatilla$ $koreana$. The increase immune cells in the high dose group of $Pulsatilla$ $koreana$ might immunomodulating activity. No significant differences in body weight, feed intake, serum biochemical analysis and histopathological between control and fed group were found. In conclusion, $Pulsatilla$ $koreana$ and $Artemisiae$ $annuae$ is physiologically safe and improve immunomodulating activity.

• Journal of Nutrition and Health
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• v.43 no.2
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• pp.105-113
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• 2010

Ixeris dentata var. albiflora Nakai, a herbal plant, is often used to make a strong stomach as an antiphlogistic used when dyspepsia and to improve appetite in Korea and China. And also it is used for adult diseases such as diabetes and liver diseases as Korean traditional medicine. In this study, the composition and DPPH radical scavenging activities of the root of Ixeris dentata var. albiflora Nakai and its effects on cell viability on vero and chang cells were investigated. Moisture, crude ash, crude protein and crude lipid were 79.14, 2.49, 8.28 and 2.56 g/100 g respectively. The highest mineral content was K. The major free sugars were glucose, fructose and sucrose. Major fatty acid are linoleic acid, palmic acid and linolenic acid. Major amino acids were glutamic acid, arginine and aspartic acid and the total contents of amino acids were 28.12 mg/g. The methanol extracts were further fractionated with n-hexane, chloroform, ethylacetate, butanol and water to get an active fraction. In addition, cell viabilities in each fraction were determined. Methanol extract, butanol, and aqueous fraction showed strong survival rates in vero cell and chang cell viability test, and hexane, chloroform, and ethylacetate fraction were examined for toxin in a cell. The root of Ixeris dentata var. albiflora Nakai had scavenging activities against DPPH radicals in a dose-dependent assay. Ethylacetate fraction's SC50 was $6.8\{\mu}g/mL$, very strong DPPH radical scavenging activities, but water fraction did not show any activity.

• Applied Microscopy
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• v.29 no.3
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• pp.363-376
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• 1999

This study aims to demonstrate the effect of chitosan oligosaccharide on the ultrastructural changes in the mouse liver toxicated by carbon tetrachloride $(CCl_4)$. A healthy male ICR mouse that weighted $27{\pm}2gm$ was used for experiment. The experimental group was divided into three groups; the group A; the pretreated group with chitosan oligosaccharide, the group B; the simultaneous group, the group C; treated only the $CCl_4$. The group A was simultaneously treated with chitosan oligosaccharide and $CCl_4$ after pretreated with chitosan oligosaccharide for 7 days. The group B injected $CCl_4$ and chitosan oligosaccharide to the intraperitoneal. The group C injected with only $CCl_4$ to the intraperitoneal. The results were as follow: In the group A, the nuclear membrane and the mitochondria were observed almost normal in shapes at overall the time. Some lamellae of the RER (rough endoplasmic reticulum) destructed until 48 hours but ribosome attached. The destructed lamellae reformed at 72 hours but the smooth membrane vesicles not observed. The lysosomes observed at 72 hours. At 96 hours, all organelles showed in normal shapes. In the group B, changes of nuclear membranes were relatively lighter than group C. Mitochondria observed normal shape through the time. Parts of RER reformed the lamellae, other parts dilated inner cavity. And lipid droplet observed around the 24 hours. Glycogen and lysosome observed 48 hours and 72 hours, respectively. In the group C, nuclear membrane was irregular and nuclear cytoplasm condensed through the time. The lamellae of RER destructed from 24 to 96 hours. Smooth membrane vesicles observed in the cytoplasm at 48 ours. Mitochondria was less effected by toxic. And from the 24 hours, the variable sizes of lipid droplets observed in tile cytoplasm. These results suggest that chitosan oligosaccharide attenuates the toxic effect of the carbon tetrachloride in the mouse liver.

• Journal of the korean academy of Pediatric Dentistry
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• v.29 no.2
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• pp.243-254
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• 2002

For the purpose of evaluating the biocompatability of 3 kinds of metallic materials frequently used in pediatric dentistry (stainless steel crown, orthodontic band, orthodontic wire), cellular and molecular studies, including cell growth and proliferation, screening of cell death with determination of types whether necrosis or apoptosis and changes in expressions of related signaling molecules were examined, using cultured human gingival fibroblasts (HGF-1), HGF-1 was cultured in Dulbecco's modified Eagle's medium. among which the 3rd to 6th generations of HGF-1 were used. The specimen were divided into stainless steel crown (R), band (B) and wire (W). The immunocytochemical study was done for the detection of anti-PCNA (proliferating cell nuclear antigen) labeling. With extracted protein, western blot was done for the detection of ERK1/2, JNK, and p38, using individual antibodies. Cultured cells proliferated, remarkably till 7 day and slightly at 11 day. There was no statistical significance in the counts of proliferating HGF-1 between control and experimental groups (p>0.05). Relative growth rates were no statistically significant difference between control and experimental groups (p>0.05). PCNA labeling indexes showing similar patterns in control and experimental groups. The expressions of ERK1 and ERK2, p38 were similar in control and experimental groups. The expression of JNK increased at 1st day, slightly decreased at 4th day and markedly increased at 7th and 11 day. Although the patterns of control and experimental groups were similar, the increased expressions of JNK at late period suggest a possible stress due to inhibited cell growth and proliferation, and worse culture condition. Conclusively, the 3 kinds of metal specimens used in this study did not induce cellular and molecular hazards during short term culture of HGF-1. But, for the better clinical stability, the establishment of long period culture and animal experiment was thought necessary.

• Korean Journal of Food Science and Technology
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• v.27 no.4
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• pp.555-563
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• 1995

This study was performed to investigate the effect of Aloe arborescens on the cadmium toxicity in rats. Thirty male Sprague-Dawley strains were divided into five groups consisting of a control group, a cadmium treatment group and 3 aloe(0.5%, 0.75%, 1%) treatment groups and observed for 9 weeks. The weight increment of the cadmium and 0.75% aloe group was higher than that of the cadmium treatment group(p<0.01). The food intake did not show the consistency rule among the experimental groups and the decrement tendency of food intake affected by cadmium feeding group. The decrement tendency of water intake affected by cadmium appeared to be suppressed by aloe treatment, especially cadmium and 0.75% aloe treatment group showed the remarkable increment of water intake. The diet efficiency of the control group was the highest among the experimental groups and that of cadmium and 0.75% aloe group was higher than other aloe treatment groups. The weight of each organ did not show consistency among the experimental groups but only the testicle of cadmium and 0.75% aloe treatment group was heavier than that of the control group. The cadmium accumulation was high in order of kidney>liver>spleen>heart>lung>testicle>brain. The cadmium content of the cadmium treatment group was more than that of cadmium and 0.5% aloe group, cadmium and 0.75% aloe group, cadmium and 1% aloe group. The cadmium content of cadmium and 0.75% aloe group was the lowest among other aloe treatment groups. Therefore, cadmium and 0.75% aloe is the most recommendable aloe treatment to eliminate the cadmium accumulated in organ.

• KSBB Journal
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• v.10 no.5
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• pp.561-567
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• 1995

Using superoxide dismutase (SOD)-deficient mutants of Saccharomyces cerevisiae, the oxygen toxicity induced by paraquat was studied. In aerobic culture condition, yeasts lacking MnSOD (milochondrial SOD) showed more significant growth retardation than CuZnSOD (cytoplasmic SOD)-deficient yeasts. However, not so big differences in growth pattern of those mutants compared with wild type were observed under anaerobic condition. When exposed to paraquat, the growth of yeasts lacking CuZnSOD was severely affected by higher than 0.01mM of paraquat in culture medium. By the analysis of several cellular components ivolved in free radical generating and scavenging system, it was found that, under aerobic condition, the content of lipid peroxides in cell membrane as well as cellular activity of glutathion peroxidase of CuZnSOD-deficient mutants was increased in the presence of paraquat, although significant decrease of catalase activity was observed in those stratns. In MnSOD-deficient yeast, however, increment in cellular activity of glutathion peroxldase and catalase by paraquat was observed without any deterioration of membrane lipid. It implies that the lack of mitochondrial SOD could be compensated by both of glutathion peroxldase and catalase, but that only glutathion peroxidase might act for CuZnSOD in cytoplasm. In contrast, all of SOD-deficient mutants showed a significant decrease in catalase activity, but slight increase in the activities of glutathion peroxidase, when cultivated anaerobically in the medium containing paraquat. Nevertheless, any significant changes of lipid peroxides in cell membranes were not observed during anaerobic cultivation of SOD-deficient mutants. It suggests that a little amount of free radicals generated by paraquat under anaerobic condition could be sufficiently overcome by glutathion peroxidase but not by catalase.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.9
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• pp.1370-1377
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• 2013

Rice is widely grown in Asia and is one of the major dietary staples in the world. Also, rice contains antioxidants which can prevent from oxidative stress related diseases, including cancer, atherosclerosis, and diabetes. Because the rice is consumed cooked, the effect of the cooking process on the antioxidative and antigenotoxic properties of rice is lacking. The aim of this study was to determine the effects of cooking on the antioxidant and antigenotoxic effects of white rice (WR), brown rice (BR), and germinated brown rice (GBR). The antioxidant activities were measured for total phenolic content (TPC), DPPH radical scavenging activity (DPPH RSA), total antioxidant capacity (TRAP), and oxygen radical absorbance capacity (ORAC). The highest TPC was found in uncooked BR (18.4 mg gallic acid equivalent/100 g). After cooking, the TPC of WR significantly increased, while the TPC of BR and GBR were reduced by 47.7% and 36.7%, respectively. The $IC_{50}$ for DPPH RSA was not significantly different in uncooked rice, while the DPPH RSA of WR and GBR decreased after cooking and the DPPH RSA of BR significantly increased. TRAP values in BR and GBR increased after cooking, while the value of WR decreased. The ORAC values of uncooked WR, BR, and GBR were 5.3, 4.3, and $3.9{\mu}M$ trolox equivalent at the concentration of $50{\mu}g/mL$. After cooking, the ORAC value of BR remained unchanged, while the value of GBR increased and the value of WR decreased. The antigenotoxic activities of WR, BR, and GBR were determined by measuring the inhibitory effects of $H_2O_2$-induced DNA damage on human leukocytes using the comet assay. The results showed that all rice tested showed a significant antigenotoxic effect against oxidative stress, except for the cooked white rice. Overall, our results indicate the addition of brown rice and/or germinated brown rice to cooked white rice is a good option for improving the benefits of rice.