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Superoxide Dismutase Gene Expression Induced by Lipopolysaccharide in Alveolar Macrophage of Rat (폐포대식세포에서 내독소 자극에 의한 Superoxide Dismutase 유전자발현의 조절 기전)

  • Park, Kye-Young;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Hyun, In-Gyu
    • 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.

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The Change of Antioxidant Enzyme (Superoxide Dismutase, Catalase, Glutathione Peroxidase) in the Endotoxin Infused Rat Lung (내독소 투여후 쥐의 폐조직내 Antioxidant (Superoxide Dismutase, Catalase, GSH-Peroxidase)의 변화에 대한 연구)

  • Song, Jeong-Sup;Kim, Chi-Hong;Kwon, Soon-Seog;Kim, Young-Kyoon;Kim, Kwan-Hyoung;Han, Ki-Don;Moon, Hwa-Sik;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.2
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    • pp.104-111
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    • 1993
  • Background: Gram-negative bacterial endotoxin induced septicemia is known to be a leading cause in the development of adult respiratory distress syndrome(ARDS). The mechanism of endotoxin induced lung injury is mainly due to the activated neutrophils which injure the capillary endothelial cells by releasing oxidant radical and resulted in pulmonary edema. We studied the change of antioxidant enzyme in the case of large or small, intermittant dose of endotoxin infused rat lungs. Methods: Endotoxin was given to the rat through the peritoneal cavity in the dose of 7 mg/kg body weight in the large dose group and 1 mg/kg for 10 days in the small dose group. Bronchoalveolar lavage (BAL) was done and rats were killed at 6, 12, 24 hours after single endotoxin injection in the large dose group and 3, 7, 10 days after daily endotoxin injection for 10 days in the small dose group. The lungs were perfused with normal saline through the pulmonary artery to remove the blood and were homogenized in 5 volume of 50 mM potassium phosphate buffer containing 0.1 mM EDTA. After centrifuging at 100,000 g for 60 minute, the supernatent was removed and stored at $-70^{\circ}C$ until measuring for superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-Px) and protein. Results: We observed the following results. 1) The lung wet/dry weight ratio and albumin concentration in the BAL fluids were increased to peak at 12 hours and neutrophil number in the BAL fluids were peak at 6 hours after endotoxin injection in the large dose group. 2) Cu, Zn SOD (IU/mg protein) was significantly decreased after 6, 12 hours after endotoxin injection in the large dose group. 3) There were no singnificant change in the level of Mn SOD, catalase, GSH-Px after endotoxin injection in both groups. Conclusion: Endotoxin in the large dose group produced the acute pulmonary edema and decreased the Cu, Zn SOD in the lung tissue after injecting endotoxin at 6 and 12 hours. These phenomenon may be due to the cell membrane damage by endotoxin. Further research would be necessary whther giving SOD by intratracheal route or method to increase the synthesis of SOD may lessen the acute lung injury by endotoxin.

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The Effects of Mycobacterium Tuberculosis on Alveolar Macrophages -The Alterations of Superoxide Production in both Human and Rat Alveolar Macrophages Exposed to Mycobacterium Tuberculosis H37Ra Strain- (결핵균이 폐포대식세포의 기능에 미치는 영향에 관한 연구 -H37Ra 결핵균종에 의한 사람 몇 백서 폐포대식세포의 Superoxide 생성의 변화-)

  • Kim, Keon-Youl;Lee, Kye-Young;Hyun, In-Kyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.6
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    • pp.526-535
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    • 1992
  • Background: The oxygen radicals released by alveolar macrophages contribute to killing of microorganisms including M. tuberculosis. Macrophages are "primrd" for enhanced oxygen radical release by macrophage activator like IFN-$\gamma$ and LPS, which do not themselves cause release of oxygen radicals. Actural production of oxygen radicals is "triggered" by phagocytosis or by exposure to chemical stimuli like PMA or FMLP. There has been debates about the priming effect of alveolar macro phages because they are exposed to usual environmental particles unlike blood monocytes. Therefore we examined priming effect of IFN-$\gamma$ in human alveolar macrophages comparing with that in blood monocytes and rat alveolar macrophages. And we observed the alterations of superoxide production in both human and rat alveolar macrophages after exposure to M. tuberculosis H37Ra bacilli itself and its lysate. Methods: Bronchoalveolar lavage fluid was processed to isolate alveolar macrophages by adherence and the adherent cells were removed by cold shock method. After exposure to M. tuberculosis H37Ra strain, alveolar macrophages were incubated for 24 hours with IFN-$\gamma$. The amount of superoxide production stimulated with PMA was measured by ferricytochrome C reduction method. Results: 1) The priming effect in human alveolar macrophages was not observed even with high concentration of IFN-$\gamma$ while it was observed in blood monocytes and rat alveolar macrophages. 2) Both human and rat alveolar macrophages exposed to avirulent H37Ra strain showed triggering of superoxide release and similar results were shown with the exposure to H37Ra lysate. Conclusion: The priming effect in human alveolar macrophages is not observed because of its usual exposure to environmental particles and avirulent H37Ra strain does not inhibit the activation of alveolar macrophages.

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The Effect of Vitamin E on the Composition of inflammatory Cells in Alveoli after Paraquat Intoxication in Rats (Paraquat에 의한 급성 폐손상에서 Vitamin E처치가 기관지폐포 세척액내 세포조성에 미치는 영향)

  • Song, Kwang-Seon;Lee, Won-Yeon;Cho, Do-Yeun;Yong, Suk-Joong;Shin, Kye-Chul
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.6
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    • pp.1332-1342
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    • 1997
  • Background : Acute pulmonary injury by paraquat are caused by multiple mechanisms including direct injury with oxygen free radicals and several mediators released from inflammatory cells. In order to clarify whether vitamin E could reduce tissue damages induced by intraperitoneal administration of paraquat and to investigate the pathogenetic mechanisms of paraquat-induced pulmonary injury, vitamin E as a free radical scavenger was administered. Method : Rats were divided into three groups (group 1 : control, group 2 : paraquat treated group, group 3 : paraquat and vitamin E treated group). Animals were sacrificed on day 1, day 2, day 3, and day 8 after the administration of saline, paraquat, or paraquat/vitamin E. Results : Treatment with vitamin E decreased the death rate of rats treated with paraquat. Comparing with control group ($1.37{\times}10^6/ml$), mean total cell counts recovered from the lavage fluid from animals treated with paraquat($1.65{\times}10^6/ml$) were increased(p=0.06). Magnitudes of increment of the total cell counts on the Day 8 in the vitamin E treated group were smaller than those of the animals treated with paraquat alone. The neutrophils began to appear in significant amounts in the lavage fluid on Day 8 after the administration of paraquat(37.0+12.7%). A significant decreasing neutrophil concentration at Day 8 was observed in the paraquat/vitamin E treated group(20.6+13.4%). Histologically the degree of pulmonary fibrosis was most prominent in the paraquat treated group while diffuse alveolar damage was continuously observed in the paraquat/vitamin E treated group and extensive interstitial lymphocytic infiltration was seen in the paraquat/vitamin E treated group. The paraquat/vitamin E treated group showed the less histologic changes. Conclusion : In this study vitamin E acting as a scavenger of neutrophil-derived free radicals and suppressant of lipid peroxidation, seemed to be the effective antioxidant in the inhibition of paraquat-induced pulmonary injury.

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Adhesion-induced generation of oxygen free radical from human alveolar macrophages and its mechanisms (폐포대식세포의 부착에 의한 산소유리기 분비능 활성화 및 그 기전)

  • Chung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.2
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    • pp.210-220
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    • 1996
  • Background : Neutrophils or monocytes separated in vitro by the adherence to plastic surface are known to be activated by surface adherence itself and subsequent experimental data might be altered by surface adherence. In the process of surface adherence, adhesion molecules have a clear role in intracellular signal pathway of cellular activation. Human alveolar macrophages(HAM) are frequently purified by the adherence procedure after bronchoalveolar lavage. But the experimental data of many reports about alveolar macrophages have ignored the possibility of adhesion-induced cellular activation. Method : Bronchoalveolar lavage was performed in the person whose lung of either side was confirmed to be normal by chest CT. With the measurement of hydrogen peroxide release from adherent HAM to plastic surface and non-adherent HAM with or without additional stimulation of phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine (fMLP), we observed the effect of the adherence to plastic surface. We also evaluated the effect of various biological surfaces on adhesion-induced activation of HAM. Then, to define the intracellular pathway of signal transduction, pretreatment with cycloheximide, pertussis toxin and anti-CD11/CD18 monoclonal antibody was done and we measured hydrogen peroxide in the culture supernatant of HAM. Results : 1) The adherence itself to plastic surface directly stimulated hydrogen peroxide release from human alveolar macrophages and chemical stimuli such as phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine(fMLP) colud not increase hydrogen peroxide release in these adherent macrophages which is already activated. 2) PMA activated human alveolar macrophages irrespective of the state of adhesion. However, fMLP stimulated the release of hydrogen peroxide from the adherent macrophages, but not from the non-adherent macrophages. 3) HAM adherent to A549 cell(type II alveolar epithelium-like human cell line) monolayer released more hydrogen peroxide in response to both PMA and fMLP. This adherence-dependent effect of fMLP was blocked by pretreatment of macrophages with cycloheximide, pertussis toxin and anti-CD18 monoclonal antibody, Conclusion : These results suggest that the stimulatory effect of PMA and fMLP can not be found in adherent macrophage because of the activation of human alveolar macrophage by the adherence to plastic surface and the cells adhered to biologic surface such as alveolar epithelial cells are appropriately responsive to these stimuli. It is also likely that the effect of fMLP on the adherent macrophage requires new protein synthesis via G protein pathway and is dependent on the adhesion between alveolar macrophages and alveolar epithelial cells by virtue of CD11/CD18 adhesion molecules.

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Ser326Cys Polymorphism of hOGG1 Gene and Risk of Primary Lung Cancer in Koreans (한국인에서 hOGG1 유전자의 Ser326Cys 다형성과 원발성 폐암의 위험도)

  • Chae, Sang-Chul;Kim, Kyung-Rock;Joo, So-Young;Lee, Su-Yean;Kang, Kyung-Hee;Jeon, Kyung-Neoyh;Cha, Seung-Ick;Kim, Chang-Ho;Jung, Tae-Hoon;Park, Jae-Yang
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.1
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    • pp.5-13
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    • 2002
  • Background: DNA repair plays a crucial role in protecting the genome from cancer-causing agents. Therefore, a reduced DNA repair capacity can increase the susceptibility to cancer. The human OGG1 (hOGG1) gene encodes DNA glycosylase/apurinic lyase and excise 8-hydroxyguanine, one of the major premutagenic DNA lesions, which is produced by oxygen radical forming agents including smoking. Recently several polymorphisms in the hOGG1 gene were identified, and it is possible that these polymorphism') may affect the DNA repair capacity and thus modulate cancer susceptibility. The relationship between the codon 326 polymorphism (Ser to Cys) in the hOGG1 gene and lung cancer risk was investigated. Materials and Method: The Ser326Cys genotypes were determined using PCR-RFLP analysis in 299 primary lung cancer patients and 186 healthy controls who were frequency (case:control=3:2) matched according to age and sex. Result: The frequencies of the Ser326Cys genotypes (Ser/Ser, Ser/Cys and Cys/Cys) among cases (23.4%, 51.8%, and 24.7%, respectively) were not significantly different from those among the controls (22.6%,52.1% and 25.3%, respectively). When the analyses were stratified according to age, sex, smoking status and packyears of smoking, no significant association between this polymorphism and lung cancer risk was found. Moreover, the Ser326Cys genotype showed no apparent relationship with any of the histological types of lung cancer. Conclusion: These result suggest that the hOGG1 Ser326Cys polymorphism is not a major contributor to individual lung cancer susceptibility in Koreans.

The Opening Space for Quality of Life in South Korea (삶의 질의 공간구조화 과정에 대한 사회학적 고찰)

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    • Korea journal of population studies
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    • v.20 no.2
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    • pp.181-198
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    • 1997
  • Going beyond the previous formulations of development theories, the present paper explores the effects other than political economy on quality of life in a rapidly developing country. The major analysis takes up the historical trend and nature of the developmental transformation that is partially a consequences of state structures and partially autonomous form it in South Korea. Also, it diagnoses developmental pathways for the future track by constructing a baseline model for state transition on the basis of power game between the state and civil society in the country. The results of the historical analysis show that civil society has been transformed in the course of confrontations and interactions between the state and nationalist social movement. The distinction between developmental(or bureaucratic authoritarian) and democratic state is presented to show that these are two qualitatively different aspects of state of state power, requiring separate analytical treatment. Furthermore, the state-centric approach which emphasizes the active role of the state at the sacrifice of societal fabric-constraining social conditions for quality of life - appears to be modified. On the contrary, the impact of civil society is transmitted both directly and indirectly via labor and ecological movement for quality of life, which is critical to the formation of the welfare state in the country. The prospect for sustainable development in Korea lies in providng and expanding quality of life in terms of the financial feasibility of the state through the public-private cooperation, and abstaining from drastic and radical commitment to welfare services as is the case with the European declines in welfare state, Further studies are needed to examine the interrelationships in different historical and cultural settings of developing counties to estimate a theory of quality of life and social justice.

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Comparative Study on Antioxidative Activity of Glycyrrhiza uralensis and Glycyrrhiza glabra Extracts by Country of Origin (원산지별 감초 추출물의 항산화 활성 비교 연구)

  • Han, Saet Byeol;Gu, Hyun A;Kim, Su Ji;Kim, Hye Jin;Kwon, Soon Sik;Kim, Hae Soo;Jeon, So Ha;Hwang, Jun Pil;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.39 no.1
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    • pp.1-8
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    • 2013
  • In this work, comparative study on antioxidative activities of extracts from Glycyrrhiza uralensis (G. uralensis) produced in Korea and in China and Glycyrrhiza glabra (G. glabra) produced in Uzbekistan was conducted. Among three origins, 50% ethanol extracts (21.15 ${\mu}g/mL$), ethyl acetate fraction (29.15 ${\mu}g/mL$) and aglycone fraction (3.26 ${\mu}g/mL$) of G. uralensis from Korea showed the higher free radical (1,1-phenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$) than extracts from other origins. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of extracts from three origins on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using luminol-dependent chemiluminescence assay 50% ethanol extract (1.00 ${\mu}g/mL$) and ethyl acetate fraction (0.34 ${\mu}g/mL$) of G. uralensis from China showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of G. uralensis and G. glabra extracts on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. 50% ethanol extract and aglycone fraction of G. uralensis and G. glabra extracts from three origins showed cellular protective effects in a concentration dependent manner (5 ~ 50 ${\mu}g/mL$). Aglycone fraction of G. uralensis from Korea (${\tau}_{50}$ = 847.4 min)especially showed cellular protective effects four times higher than that from China (${\tau}_{50}$ = 194.3 min). These results indicate that G. uralensis and G. glabra extracts, which have been used as whitening agent, could be applicable to functional cosmetic ingredient as a natural antioxidant. Judging from the prominent cellular protecitve effects, it is concluded that G. uralensis and G. glabra extracts can protect the skin from $^1O_2$ and various ROS induced by UV.

Physicochemical properties of Salvia miltiorrhiza Bunge following treatment with enzymes (효소 처리에 따른 단삼 추출물의 이화학적 특성)

  • Kim, Sun-Hwa;Hwang, In-Wook;Chung, Shin-Kyo;Seo, Young-Jin;Kim, Jong-Soo;Jeong, Yong-Jin;Kim, Mi-Yeon
    • Food Science and Preservation
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    • v.22 no.5
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    • pp.699-707
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    • 2015
  • To improve the utilization of the domestic plant Salvia miltiorrhiza Bunge (Danshen), this study investigated changes in the physicochemical qualities of Danshen extracts obtained from low-temperature extraction using the enzymes amylase, cellulase, pectinase, and protease. Changes in the yield, pH, sugar content, and chromaticity were investigated. The changes were found to be highest in the amylase-treated extract with the following values: yield, 58.3%; pH, 6.04; sugar content, $5.97^{\circ}Brix$. With regard to antioxidant properties, Danshen extracts treated with amylase showed the highest DPPH and ABTS scavenging activities of 84.25% and 74.11% at 55 ppm. The total phenolic compound content was highest in the group subjected to enzyme treatment at $60^{\circ}C$. The salvianolic acid B level of the Danshen extract was the highest in the amylase-treated group, with a value of 3,002.6 mg/100 g. Cryptotanshinone level was the highest in the amylase- and protease-treated group with a value of 3.8 mg/100 g. Tanshinone I was the highest in the protease-treated group, with a value of 14.2 mg/100 g. The results showed that the indicator components of Danshen were detected as stable in the extracts after using amylase for low-temperature extraction; therefore, it would be possible to use Danshen industrially as a functional ingredient through mass production. Furthermore, the enzyme-treatment extraction could be utilized for a variety of natural products.

Effect of extract temperature and duration on antioxidant activity and sensory characteristics of Ulmus pumila bark extract (추출온도 및 시간에 따른 유백피 추출물의 항산화 활성과 음료의 관능적 특성)

  • Cho, Myoung Lae;Oh, Yu-Na;Ma, Jin-Gyeong;Lee, Su-Jin;Choi, Young-Hee;Son, Dong-Hwa;Jang, Eun Hee;Kim, Jong-Yea
    • 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.