• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.5
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• pp.545-551
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• 2017

Curcuma longa L. (CL) is widely used as a spice and coloring agent in several foods, such as curry and mustard, as well as cosmetics and drugs. In this study, we investigated the protective effects of CL extracted with various solvents [methanol (MC), ethanol (EC), acetone (AC)] on $H_2O_2-induced$ DNA damage in human leukocytes along with total polyphenol contents (TPC) and antioxidant properties. The antioxidant effects of CL were determined by measuring 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (RSA) and superoxide dismutase (SOD)-like activity. The preventive effect of CL on oxidative stress-induced DNA damage and DNA repair capacities were assessed using comet assay. MC showed the highest TPC (11.17 g gallic acid equivalents/100 g) and antioxidant properties among the solvent extracts. The $SC_{50}$ for DPPH RSA was MC: 35.0 > AC: 45.8 > EC: $57.8{\mu}g/mL$ and SOD-like activity was MC: 46.6 > EC: 141.5 > AC: $296.4{\mu}g/mL$. In the comet assay, the $ED_{50}$ value of MC showed the highest inhibition ($86.7{\mu}g/mL$) of $H_2O_2-induced$ DNA damage, followed by AC ($110.0{\mu}g/mL$) > EC ($115.8{\mu}g/mL$). Analysis of the percentage of damaged cells showed that repair capacity significantly decreased at 4, 8, and 12 h from $H_2O_2-induced$ oxidative stress in each extract. After 12 h, level of DNA damage recovery was similar to the negative control level. These results suggest that CL has potential antioxidant activity and a protective effect against oxidation-induced DNA damage, and the methanol extract of CL was the most effective.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.10
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• pp.1164-1170
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• 2017

Protaetia brevitarsis larvae (PBL) has recently been registered as a temporary food in Korea, and this study evaluated the application potential of PBL proteins as health functional food materials. Protein hydrolysates were prepared from PBL powder by enzymatic hydrolysis using five different proteases (alcalase, bromelain, flavourzyme, neutrase, and papain), and based on the results from the peptide content and SDS-PAGE analyses, PBL treated with alcalase or flavourzyme showed a high degree of hydrolysis (HD) value, whereas the HD value of those treated with neutrase, bromelain, or papain was minimal. The protein hydrolysates showing a high HD value were separated further into the fractions of >3 kDa and <3 kDa by a centrifugal filter system and then lyophilized, and according to the $RC_{50}$ values of the protein hydrolysates (<3 kDa) obtained from three different antioxidant analyses; the alcalase hydrolysates showed the highest antioxidant activity. Therefore, the alcalase hydrolysates were tested further for their inhibitory effects on the peroxidation of linoleic acid by measuring the thiobarbituric acid values. The results showed that the peroxidation of untreated linoleic acid increased dramatically during 6 days of incubation, but a pretreatment with the hydrolysates ($100{\sim}800{\mu}g/mL$) significantly inhibited the linoleic acid peroxidation in a dose-dependent manner for 6 days. Our current studies are focused on the identification of active peptide sequences from alcalase hydrolysates.

• Tuberculosis and Respiratory Diseases
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• v.41 no.3
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• pp.215-221
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• 1994

Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

• Tuberculosis and Respiratory Diseases
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• v.41 no.3
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• pp.222-230
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• 1994

Background: Paraquat, a widely used herbicide, is extremely toxic, causing multiple organ failure in humans. Paraquat especially leads to irreversible progressive pulmonary fibrosis, which is related to oxygen free radicals. However, its biochemical mechanism is not clear. Natural mechanisms that prevent damage from oxygen free radicals include changes in glutathione level, G6PDH, superoxide dismutase(SOD), catalase, and glutathione peroxidase. The authors think catalase is closely related to paraquat toxicity in the lungs Method: The effects of 3-amino-1,2,4-triazole(aminotriazole), a catalase inhibitor, on mice administered with paraquat were investigated. We studied the effects of aminotriazole on the survival of mice administered with paraquat, by comparing life spans between the group to which paraquat had been administered and the group to which a combination of paraquat and aminotriazole had been administered. We measured glutathion level, glucose 6-phosphate dehydrogenase(G6PDH), superoxide dismutase(SOD), catalase, and glutathione peroxidase(GPx) in the lung tissue of 4 groups of mice: the control group, group A(aminotriazole injected), group B(paraquat administered), group C(paraquat and aminotriazole administered). Results: The mortality of mice administered with paraquat which were treated with aminotriazole was significantly increased compared with those of mice not treated with aminotriazole. Glutathione level in group B was decreased by 20%, a significant decrease compared with the control group. However, this level was not changed by the administration of aminotriazole(group C). The activity of G6PDH in all groups was not significantly changed compared with the control group. The activities of SOD, catalase, and glutathione peroxidase(GPx) in the lung tissue were significantly decreased by paraquat administration(group B); catalase showed the largest decrease. Catalase and GPX were significantly decreased by aminotriazole treatment in mice administered with paraquat but change in SOD activity was not significant(group C). Conclusion: Decrease in catalase activity by paraquat suggests that paraquat toxicity in the lungs is closely related to catalase activity. Paraquat toxicity in mice is enhanced by aminotriazole administration, and its result is related to the decrease of catalase activity rather than glutathione level in the lungs. Production of hydroxyl radicals, the most reactive oxygen metabolite, is accelerated due to increased hydrogen peroxide by catalase inhibition and the lung damage probably results from nonspecific tissue injury of hydroxyl radicals.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.10
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• pp.1304-1308
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• 2006

The present study describes the preliminary evaluation of the antioxidant activity and the cytotoxic effect of Ceramium kondoi. The antioxidant activities and cytotoxic effect of the water extracts were evaluated by total phenolic contents (TPC), DPPH radical scavenging activity (RSA), reducing power (RP), comet assay, and MTT reduction assay. TPC, DPPH RSA, and RP of the extract at the concentration of $1,000{\mu}g/mL$ was $659.2{\mu}M$, 86.0%, and 1.084, respectively, and those were concentration dependent. The $200{\mu}M\;H_2O_2-induced$DNA damage was inhibited by C. kondoi water extract in a dose dependent manner in human leukocytes. The inhibition was by 62.3, 39.8, 24.8% and 16.4% at the concentration of 5, 10, $25{\mu}g/mL$ and $50{\mu}g/mL$, respectively. Cytotoxic activity on HT-29 cells and MCF-7 cells of the C. kondoi water extract at the concentration of $10{\mu}g/mL$ was 49% and 60%, respectively. These results strongly support the possibility of C. kondoi as a source of natural functional materials.

• Tuberculosis and Respiratory Diseases
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• v.42 no.4
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• pp.522-534
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• 1995

Background: In the pathogenesis of acute lung injury induced by lipopolysaccharide(LPS), oxygen radiclls are known to be involved in one part. Superoxide dismutase(SOD) protects oxygen radical-induced tissue damage by dismutating superoxide to hydrogen peroxide. In eukaryotic cells, two forms of SOD exist intracellularly as a cytosolic, dimeric copper/zinc-containing SOD(CuZnSOD) and a mitochondrial, tetrameric manganese-containing SOD(MnSOD). But there has been little information about SOD gene expression and its regulation in pulmonary alveolar macrophages(PAMs). The objective of this study is to evaluate the SOD gene expression induced by LPS and its regulation in PAMs of rat. Method: In Sprague-Dawley rats, PAMs obtained by broncholaveolar lavage were purified by adherence to plastic plate. To study the effect of LPS on the SOD gene expression of PAMs, they were stimulated with different doses of LPS($0.01{\mu}g/ml{\sim}10{\mu}g/ml$) and for different intervals(0, 2, 4, 8, 24hrs). Also for evaluating the level of SOD gene regulation actinomycin D(AD) or cycloheximide(CHX) were added respectively. To assess whether LPS altered SOD mRNA stability, the rate of mRNA decay was determined in control group and LPS-treated group. Total cellular RNA extraction by guanidinium thiocyanate/phenolfchlorofonn method and Northern blot analysis by using a $^{32}P$-labelled rat MnSOD and CuZnSOD cDNAs were performed. Results: The expression of mRNA in MnSOD increased dose-dependently, but not in CuZnSOD. MnSOD mRNA expression peaked at 8 hours after LPS treatment. Upregulation of MnSOD mRNA expression induced by LPS was suppressed by adding AD or CHX respectively. MnSOD mRNA stability was not altered by LPS. Conclusion: These findings show that PAMs of rat could be an important source of SOD in response to LPS, and suggest that their MnSOD mRNA expression may be regulated transcriptionally and require de novo protein synthesis without affecting mRNA stability.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.1 s.32
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• pp.80-98
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• 2007

Objectives : This study was carried out to evaluate anti-oxydative, anti-tumor effect for clinical application of Whakijogyungtang (WJT) Results : 1. DPPH radical scavenging activities of WJT water extracts(Exts) were in proportion as concentration of WJT.(3 ${\mu}g/ml:12.6{\pm}2.3$ %) 2. ABTS+ scavenging activities of WJT water Exts were more effective in high density.(3 ${\mu}g/ml:4.3{\pm}1.6$ %, 10 ${\mu}g/ml$: $11.8{\pm}2.5$ %, 30 ${\mu}g/ml:45.3{\pm}3.2%$ 100 ${\mu}g/ml$: $62.7{\pm}4.8%$) 3. Hydrogen peroxide$(H_2O_2)$ scavenging activities of WJT water Exts were effective.(3 ${\mu}g/ml:4.7{\pm}0.8$ %, 10 ${\mu}g/ml: $8.2{\pm}1.6$ %, 30 ${\mu}g/ml:19.5{\pm}3.2$ % 100 ${\mu}g/ml$: $24.6{\pm}3.8$ %) 4. Anti oxidative effects against linoleic acid were not effective. 5. The generation of $O_2\;^-$ in S-180 cells were according to the concentration of WJT water Exts, specially effective over 100 ${\mu}g/ml$ concentration. 6. The SOD activities in S-180 cells were in proportion as cytotoxicity against S-180 cells of WJT water Exts. 7. The GPx activities in S-180 cells were in proportion as cytotoxicity against S-180 cells of WJT water Exts(more effective on 300 ${\mu}g/ml$ and 1000 ${\mu}g/ml$ concentration), but the catalase activities in S-180 cells were not effective. 8. The results of activites against multi-drug-resistance(MDR) of WJT were as follows. 1) In water Exts from WJT, cytotoxicity against AML-2/D100 with vincristine($IC_{50:}39.78$ ${\mu}g/ml$) was more effective than without vincristine($IC_{50:}$ 183.58 ${\mu}g/ml$), Cross resistance(CR:3.85) was not effective, and anti-MDR activites(RF) was effective.(RF:3.85) 2) In hexane fraction, cytotoxicity against AML-2/D100 with vincristine ($IC_{50:}130.88$ ${\mu}g/ml$) was more effective than without vincristine ($IC_{50:}293.10$ ${\mu}g/ml$) and anti-MDR activites(RF) was effective.(RF:4.61) 3) In chloroform fraction, the cytotoxicity against AML-2/WT and AML-2/D100 was not effective. 4) In ethyl acetate fraction, cytotoxicity against AML-2/D100 with vincristine($IC_{50:}36.43$ ${\mu}g/ml$) was more effective than without vincristine ($IC_{50:}73.07$ ${\mu}g/ml$), Cross resistance(CR:0.53) was not effective, and anti-MDR activites(RF) was effective.(RF:2) 5) In butanol fraction, the cytotoxicity against AML-2/WT and AML-2/D100 was not effective. 6) In $H_2O$ fraction, the cytotoxicity against AML-2/WT and AML-2/D100 was not effective. Conclusion : These result suggest that WJT has antioxidative effects, anti-tumor effects by apoptosis of free radical$(O_2\;^-)$ activity, and anti-MDR activites(especially hexane and ethyl acetate fraction).

• Food Science and Preservation
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• v.23 no.7
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• pp.995-1003
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• 2016

Ulmus pumila L. bark underwent distilled water extraction under three temperature condition ($4^{\circ}C$, room temperature, or $80^{\circ}C$) and two extraction times (1, or 5 min) in order to develop a functional beverage products. Changes in yield, pH, color, total phenolic (TP) content, tannin content and antioxidant activity of the aqueous extracts were evaluated for each extraction temperature and duration. Extraction conditions did not affect yield or pH value of the extracts; however CIE $b^*$ values were high in extracts prepared under high extraction temperature ($80^{\circ}C$) and long extraction duration (5 min) conditions. Both extraction temperature and duration affected the TP and tannin contents of the extracts; however, all extraction conditions resulted in ${\geq}450\;mg\;GAE/g$ TP content and ${\geq}80\;mg\;CE/g$ tannin content. All extracts exhibited ABTS and DPPH radical scavenging ability similar to that of vitamin C. Nitric oxide inhibition activity was lower in the 5 min duration sample than in the 1 min sample. The $4^{\circ}C$ extraction temperature produced an extract with the highest reducing power and hydrogen peroxide values. Extraction temperature also affected sensory evaluation results with the $80^{\circ}C$ extraction temperature producing significantly higher flavor, bitterness, and color score, than those obtained under $4^{\circ}C$ and room temperature extraction conditions.

• Korean Journal of Microbiology
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• v.52 no.2
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• pp.140-147
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• 2016

A process for manufacturing virally-safe porcine bone hydroxyapatite (HA) has been developed to serve as advanced xenograft material for dental applications. Porcine bone pieces were defatted with successive treatments of 30% hydrogen peroxide and 80% ethyl alcohol. The defatted porcine bone pieces were heat-treated in an oxygen atmosphere box furnace at $1,300^{\circ}C$ to remove collagen and organic compounds. The bone pieces were ground with a grinder and then the bone powder was sterilized by gamma irradiation. Morphological characteristics such as SEM (Scanning Electron Microscopy) and TEM (Transmission Electron Microscopy) images of the resulting porcine bone HA (THE Graft$^{(R)}$) were similar to those of a commercial bovine bone HA (Bio-Oss$^{(R)}$). In order to evaluate the efficacy of $1,300^{\circ}C$ heat treatment and gamma irradiation at a dose of 25 kGy for the inactivation of porcine viruses during the manufacture of porcine bone HA, a variety of experimental porcine viruses including transmissible gastroenteritis virus (TGEV), pseudorabies virus (PRV), porcine rotavirus (PRoV), and porcine parvovirus (PPV) were chosen. TGEV, PRV, PRoV, and PPV were completely inactivated to undetectable levels during the $1,300^{\circ}C$ heat treatment. The mean log reduction factors achieved were $${\geq_-}4.65$$ for TGEV, $${\geq_-}5.81$$ for PRV, $${\geq_-}6.28$$ for PRoV, and $${\geq_-}5.21$$ for PPV. Gamma irradiation was also very effective at inactivating the viruses. TGEV, PRV, PRoV, and PPV were completely inactivated to undetectable levels during the gamma irradiation. The mean log reduction factors achieved were $${\geq_-}4.65$$ for TGEV, $${\geq_-}5.87$$ for PRV, $${\geq_-}6.05$$ for PRoV, and $${\geq_-}4.89$$ for PPV. The cumulative log reduction factors achieved using the two different virus inactivation processes were $${\geq_-}9.30$$ for TGEV, $${\geq_-}11.68$$ for PRV, $${\geq_-}12.33$$ for PRoV, and $${\geq_-}10.10$$ for PPV. These results indicate that the manufacturing process for porcine bone HA from porcine-bone material has sufficient virus-reducing capacity to achieve a high margin of virus safety.

• Journal of Embryo Transfer
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• v.30 no.1
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• pp.7-15
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• 2015

The objective of this study was to investigate the efficiency of nicotinic acid on sperm cryosurvival and fertilization ability in frozen-thawed boar semen. Boar semen was collected by glove-hand method and was frozen using freezing solution treated to 0, 5, 10 and 20 mM of nicotinic acid. The frozen sperm for sperm characteristic analysis was thawed such as viability, acrosome reaction, and mitochondrial integrity. The frozen-thawed sperm was estimated by SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction and Rhodamine123/PI double staining for mitochondrial integrity using a flow cytometry. The embryo was estimated in vitro development and DCFDA staining for reactive oxygen species assessment. As results, frozen-thawed sperm viability was significantly higher in 5 and 10 mM ($61.1{\pm}1.5%$,$64.7{\pm}2.0%$) of nicotinic acid than other groups (0 mM, $52.1{\pm}2.3%$; 20 mM, $47.8{\pm}5.1%$, P<0.05). The live sperm with acrosome reaction was significantly higher in 5 and 10 mM of nicotinic acid ($26.1{\pm}1.8%$, $24.9{\pm}1.5%$) than other groups (0 mM, $35.3{\pm}0.8%$; 20 mM, $36.5{\pm}1.9%$, P<0.05). The live sperm with mitochondrial integrity was significantly higher in 5 and 10 mM ($84.2{\pm}3.6%$, $88.4{\pm}2.3%$) of nicotinic acid than other groups (0 mM, $77.3{\pm}4.4%$; 20 mM, $73.3{\pm}3.6%$, P<0.05). Blastocyst rate of in vitro development was significantly higher in 10 mM ($17.0{\pm}1.3%$) of nicotinic acid than other groups (0 mM, $9.4{\pm}0.5%$; 5mM, $12.6{\pm}0.8%$; 20 mM, $5.0{\pm}1.0%$, P<0.05). Moreover, total cell number was higher in 5 and 10 mM ($53.6{\pm}2.9%$, $57.9{\pm}2.8%$) of nicotinic acid than other groups (0 mM, $41.0{\pm}1.4%$; 20 mM, $23.2{\pm}2.8%$, P<0.05). Hydrogen peroxide in embryos was lower in 5 mM nicotinic acid ($0.7{\pm}0.1%$) than other groups (0 mM, $1.0{\pm}0.1%$; 10mM, $0.9{\pm}0.0%$; 20 mM, $1.4{\pm}1.0%$, P<0.05). In conclusion, nicotinic acid-treated semen improves cryosurvival and quality of spermatozoa. Also, the fertilized oocytes with nicotinic acid improve quality of embryo and blastocyst formation.