• Tuberculosis and Respiratory Diseases
• /
• v.51 no.4
• /
• pp.315-324
• /
• 2001

Background : IL-8 is a potent chemotactic cytokine that plays an important role in the host defense mechanism against M. tuberculosis by recruiting inflammatory cells to the site of the infection. Lung epithelial cells, as well as alveolar macrophages are known to produce IL-8 in response to M. tuberculosis. IL-8 gene expression is mainly regulated on the level of transcription by NF-${\kappa}B$. This study investigated whether or not A549 cells produce IL-8 in NF-${\kappa}B$ dependent mechanism in response to macrophages phagocytosing M. tuberculosis. Methods : Peripheral blood monocytes that were obtained from healthy donors were cultured for 24 h with M. tuberculosis and a conditioned medium(CoMTB) was obtained. As a negative control, the conditioned medium without M. tuberculosis (CoMCont) was used. A549 cells were stimulated with M. tuberculosis, CoMCont and CoMTB and the IL-8 concentration in the culture media was measured by ELISA. The CoMTB induced IL-8 mRNA expression in the A549 cells was evaluated using RT-PCR, and CoMTB induced $I{\kappa}B{\alpha}$ degradation was measured using western blot analysis. CoMTB induced nuclear translocation and DNA binding of NF-${\kappa}B$ was also examined using an electrophoretic mobility shift assay(EMSA), and the CoMTB induced NF-${\kappa}B$ dependent IL-8 transcriptional activity was measured using a luciferase reporter gene assay. Results : CoMTB induced IL-8 production by A549 cells($46.8{\pm}4.8\;ng/ml$) was higher than with direct stimulation with M. tuberculosis ($6.8{\pm}2.9\;ng/ml$). CoMTB induced IL-8 mRNA expression increased after 2 h of stimulation and was sustained for 24 h. $I{\kappa}B{\alpha}$ was degraded after 10 min of CoMTB stimulation and reappeared by 60 min. CoMTB stimulated the nuclear translocation and DNA binding of NF-${\kappa}B$. The CoMTB induced NF-${\kappa}B$ dependent IL-8 transcriptional activity($13.6{\pm}4.3$ times control) was higher than either CoMCont($2.0{\pm}0.6$ times control) or M. tuberculosis ($1.4{\pm}0.6$ times control). Conclusion : A conditioned medium of peripheral blood monocytes phagocytosing M. tuberculosis stimulates NF-${\kappa}B$ dependent IL-8 production by the lung epithelial cells.

• Journal of Life Science
• /
• v.20 no.2
• /
• pp.281-291
• /
• 2010

We investigated the antioxidant effects of hederagenin 3-O-b-D-glucopyranosyl($1{\rightarrow}3$)-a-L-rhamnopyranosyl($1{\rightarrow}2$)-a-L-arabinopyranoside (HDL) isolated from root bark of Ulmus davidiana on the activity of enzymes related to reactive oxygen species (ROS) in human osteosarcoma U2OS cells. Cobalt chloride ($CoCl_2$), a transition metal, was used as an inducer of oxidative stress, generating hydrogen peroxide ($H_2O_2$) via increasing xanthine oxidase (XO) activity. The increased levels of $H_2O_2$, XO, ferritin, and ferritin iron by $CoCl_2$ were diminished effectively by co-treatment with HDL in U2OS cells. Furthermore, decreased levels of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) by $CoCl_2$ were highly increased by co-treatment with HDL in U2OS cells; however, the levels of glutathione peroxidase (GPx) did not change. The increased contents of TBARS related to lipid peroxidation were significantly reduced by HDL in U2OS cells. The concentration of GSH changed in a pattern that went against regulated TBARS by $CoCl_2$ and HDL. We examined the expression of p53, $p21^{CIP1/WAF1}$, and $p27^{KIP1}$ proteins related to oxidative stress and cell cycle regulation. As a result, the expression of $p27^{KIP1}$ modulated by $CoCl_2$ was not changed by HDL. However, the expression of p53 and $p21^{CIP1/WAF}$ increased by $CoCl_2$ was reduced by HDL in U2OS cells. Together with alteration of p53 and $p21^{CIP1/WAF1}$ proteins, the accumulated cells at G1 phase by $CoCl_2$ was decreased by HDL in U2OS cells. Our data suggests that HDL inhibits $CoCl_2$-generated ROS in U2OS cells, providing potentially new antioxidant compounds that are isolated from natural products.

• The Korean Journal of Food And Nutrition
• /
• v.17 no.3
• /
• pp.237-245
• /
• 2004

The physicochemical and physiological activities of domestic garlic from 3 different areas (Namhae, Jeju and Uiseong) were analyzed. The contents of moisture, ash, crude protein and crude fiber in garlic were little different in 3 kinds of area. Total sugar and water soluble phenolic compounds were higher in garlic from Namhae. The free sugars found in garlic were fructose, sucrose and lactose. Five kinds of organic acids were determined. Malonic acid and citric acid contents in garlic from Namhae were 23.7${\pm}$1.16 mg% and 22.1${\pm}$0.82 mg%, respectively. Total mineral content of garlic samples were in a range of 7112.6～9067.3 mg%, the potassium content showed the highest concentration (4117.3${\pm}$7.19～5175.3${\pm}$9.61 mg%). The electron donating abilities in 0.2% of garlic from Namhae and Uiseong showed 46.2${\pm}$1.25% and 37.0${\pm}$1.l6%, respectively. The nitrite scavenging effect was measured at different conditions (pH 1.2 and 4.2). The nitrite scavenging effects were higher at pH 1.2, and reached more than 95% by adding 0.2% and 0.1 % of garlic juice at pH 1.2. Addition 0.02～0.001 % garlic juice in showed the SOD-like activities. Its activity of garlic from Namhae. was a range of 6.0${\pm}$0.37～14.4～0.69%. It was found that 0.2% and 0.1% garlic showed strong antimicrobial action against growth of all the tested bacteria. Antimicrobial action. was showed 74.7${\pm}$0.70% and 51.7${\pm}$1.l1% on Sal. typhimurium in 0.2% of garlic from Namhae and Uiseong and 28.6${\pm}$0.90% on B. subtilis in garlic from Jeju.

• Korean journal of food and cookery science
• /
• v.24 no.6
• /
• pp.905-911
• /
• 2008

Dried and roasted Gugija (Lycii fructus) were extracted with water, 50% ethanol and 100% ethanol, after which the physico-chemical properties of the extracts were evaluated. The extraction yield was higher when using water for the extraction solvent than when the other solvents were used, while the water extract of roasted Gugija had the highest yield. Furthermore the pH of the extracts increased as the ethanol concentration increased, and the pH of dried Gugija was higher than that of roasted Gugija when extracted using the same extraction solvent. The sugar concentrations of the extracts from dried and roasted Gugija were $15.0{\sim}20.1\;%Brix$ and $18.0{\sim}21.2\;%Brix$, respectively. The total polyphenol contents of the extracts from the dried and roasted Gugija were $6.9{\sim}19.0\;mg/g$ and $12.4{\sim}15.8\;mg/g$, respectively. Dried Gugija extract with water had a higher the total polyphenol contents than the other extract. The total polyphenol contents of roasted Gugija extracts were higher than those of dried Gugija, when using 50% or 100% ethanol for extraction solvent. The electron donating ability and total antioxidant activity of dried Gugija were $67.6{\sim}87.7%$ and $58.6{\sim}85.0%$, respectively, whereas those of roasted Gugija were $84.7{\sim}89.8%$ and $80.6{\sim}83.7%$, respectively. Dried and roasted Gugija extracts were higher electron donating ability and total antioxidant activity, when using water, and 50% or 100% ethanol, respectively. The predominant amino acid in all extracts was threonine. The essential amino acids constituted approximately $44.9{\sim}63.6%$ and $45.4{\sim}59.0%$ of the total amino acids of extracts from the dried and roasted Gugija, respectively. Finally, the total polyphenol contents and antioxidant activities showed that optimal extraction solvent would be water, and 50% or 100% ethanol for dried and roasted Gugija, respectively.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 2017.06a
• /
• pp.194-194
• /
• 2017

Camelina (Camelina sativa L.) is a potential bio-energy crop that has short life cycle about 90 days and contains high amount of unsaturated fatty acid which is adequate to bio-diesel production. Enhancing environmental stress tolerance is a main issue to increase not only crop productivity but also big mass production. CsRCI2s (Rare Cold Inducible 2) are cold and salt stress related protein that localized at plasma membrane (PM) and assume to be membrane potential regulation factor. These proteins can be divide into C-terminal tail (CsRCI2D/E/F/G) or no-tail group (CsRCI2A/B/C/H). However, function of CsRCI2s are less understood. In this study, physiological responses and functional characterization of CsRCI2s of Camelina under salt stress were analyzed. Full-length CsRCI2s (A/B/E/F) and CsPIP2;1 sequences were confirmed from Camelina genome browser. Physiological investigations were carried out using one- or four-week-old Camelina under NaCl stress with dose and time dependent manner. Transcriptional changes of CsRCI2A/B/E/F and CsPIP2;1 were determined using qRT-PCR in one-week-old Camelina seedlings treated with NaCl. Translational changes of CsRCI2E and CsPIP2;1 were confirmed with western-blot using the antibodies. Water transport activity and membrane potential measurement were observed by cRNA injected Xenopus laevis oocyte. As results, root growth rate and physiological parameters such as stomatal conductance, chlorophyll fluorescence, and electrolyte leakage showed significant inhibition in 100 and 150 mM NaCl. Transcriptional level of CsPIP2;1 did not changed but CsRCI2s were significantly increased by NaCl concentration, however, no-tail type CsRCI2A and CsRCI2B increased earlier than tail type CsRCI2E and CsRCI2F. Translational changes of CsPIP2;1 was constitutively maintained under NaCl stress. But, accumulation of CsRCI2E significantly increased by NaCl stress. CsPIP2;1 and CsRCI2A/B/E/F co-expressed Xenopus laevis oocyte showed decreased water transport activity as 61.84, 60.30, 62.91 and 76.51 % at CsRCI2A, CsRCI2B, CsRCI2E and CsRCI2F co-expression when compare with single expression of CsPIP2;1, respectively. Moreover, oocyte membrane potential was significantly hyperpolarized by co-expression of CsRCI2s. However, higher hyperpolarized level was observed in tail-type CsRCI2E and CsRCI2F than others, especially, CsRCI2E showed highest level. It means transport of $Na^+$ ion into cell is negatively regulated by expression of CsRCI2s, and, function of C-terminal tail is might be related with $Na^+$ ion influx. In conclusion, accumulation of NaCl-induced CsRCI2 proteins are related with $Na^+$ ion exclusion and prevent water loss by CsPIP2;1 under NaCl stress.

• Tuberculosis and Respiratory Diseases
• /
• v.65 no.6
• /
• pp.476-486
• /
• 2008

Background: TRAIL (TNF-related apoptosis inducing ligand) is a newly identified member of the TNF gene family which appears to have tumor-selective cytotoxicity due to the distinct decoy receptor system. TRAIL has direct access to caspase machinery and induces apoptosis regardless of p53 phenotype. Therefore, TRAIL has a therapeutic potential in lung cancer which frequently harbors p53 mutation in more than 50% of cases. However, it was shown that TRAIL also could activates $NF-{\kappa}B$ in some cell lines which might inhibit TRAIL-induced apoptosis. This study was designed to investigate whether TRAIL can activate $NF-{\kappa}B$ in lung cancer cell lines relatively resistant to TRAIL-induced apoptosis and inhibition of $NF-{\kappa}B$ activation using proteasome inhibitor MG132 which blocks $I{\kappa}B{\alpha}$ degradation can sensitize lung cancer cells to TRAIL-induced apoptosis. Methods: A549 (wt p53) and NCI-H1299 (null p53) lung cancer cells were used and cell viability test was done by MTT assay. Apoptosis was confirmed with Annexin V assay followed by FACS analysis. To study $NF-{\kappa}B$-dependent transcriptional activation, a luciferase reporter gene assay was used after making A549 and NCI-H1299 cells stably transfected with IgG ${\kappa}-NF-{\kappa}B$ luciferase construct. To investigate DNA binding of $NF-{\kappa}B$ activated by TRAIL, electromobility shift assay was used and supershift assay was done using anti-p65 antibody. Western blot was done for the study of $I{\kappa}B{\alpha}$ degradation. Results: A549 and NCI-H1299 cells were relatively resistant to TRAIL-induced apoptosis showing only 20~30% cell death even at the concentration 100 ng/ml, but MG132 ($3{\mu}M$) pre-treatment 1 hour prior to TRAIL addition greatly increased cell death more than 80%. Luciferase assay showed TRAIL-induced $NF-{\kappa}B$ transcriptional activity in both cell lines. Electromobility shift assay demonstrated DNA binding complex of $NF-{\kappa}B$ activated by TRAIL and supershift with p65 antibody. $I{\kappa}B{\alpha}$ degradation was proven by western blot. MG132 completely blocked both TRAIL-induced $NF-{\kappa}B$ dependent luciferase activity and DNA binding of $NF-{\kappa}B$. Conclusion: This results suggest that inhibition of $NF-{\kappa}B$ can be a potentially useful strategy to enhance TRAIL-induced tumor cell killing in lung cancer.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.41 no.7
• /
• pp.927-935
• /
• 2012

This study was performed to develop functional vinegar by using cucumbers through two stages of fermentation. The alcohol content was maximized (7.8%) after 6-days of alcohol fermentation at $25^{\circ}C$ by adjusting the initial sugar concentration to $15^{\circ}Brix$, and vinegar with an acidity of 5.8% was obtained after 12-days of acetic acid fermentation at $30^{\circ}C$. The major sugars in the produced vinegar were glucose and fructose, which were present in concentrations of 3,067.26 and 395.73 mg%, respectively. The major organic acids were acetic acid and succinic acid, which were present in concentrations of 4,410.5 and 841.11 mg%, respectively. The total free amino acid content of the cucumber vinegar was 181.45 ${\mu}g/mL$ and citrulline, valine, aspartic acid, asparagine, and ornithine were the major amino acids. The inorganic components included various alkaline elements, such as K, Ca, and Mg. In addition, experimental methods to assess the DPPH and $ABTS^+$ radical-scavenging ability, reducing power, and ${\beta}$-carotene bleaching activity showed that the cucumber vinegar had strong antioxidant properties. The total polyphenol content, which are the major components responsible for the antioxidant activities of the cucumber vinegar, was 40.14 mg/100 mL. The cucumber vinegar showed significantly higher hepatic aldehyde dehydrogenase activity when compared to the alcoholic control (negative) and the marketing drink (positive), resulting in decreased plasma acetaldehyde concentrations in rats. These results demonstrate that cucumber vinegar possesses antioxidant properties and holds great promise for use in preventing hangovers.

• Journal of the Korean Applied Science and Technology
• /
• v.31 no.4
• /
• pp.771-780
• /
• 2014

The antioxidative effects and component analysis of the Melaleuca quinquenervia leaf extracts were investigated. All experiments were performed with 50% ethanol extract, ethyl acetate fraction and aglycone fraction obtained from dried M. quinquenervia leaves. The DPPH (1,1-phenyl-2-picrylhydrazyl) scavenging activity ($FSC_{50}$) of ethyl acetate fraction ($10.05{\mu}g/mL$) of M. quinquenervia leaf extracts was similar to (+)-${\alpha}$-tocopherol($8.89{\mu}g/mL$) known as a typical antioxidant. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of the ethyl acetate fraction ($1.61{\mu}g/mL$) and aglycone fraction ($1.07{\mu}g/mL$) of leaf extracts of M. quinquenervia on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay were similar to that of L-ascorbic acid ($1.50{\mu}g/mL$). The cellular protective effect of the extracts on the rose bengal sensitized photohemolysis of human erythrocytes was increased in a concentration dependant manner ($1{\sim}50{\mu}g/mL$). Especially, the cellular protective effects of Aglycone fraction (${\tau}_{50}=158.80min$) and 50% Ethanol extract (${\tau}_{50}=50.1{\pm}0.2min$) on the $^1O_2$-induced cellular damage of human cells were exhibited the higher than (+)-${\alpha}$-tocopherol (${\tau}_{50}=38.0min$). TLC and HPLC were used to analyse active components in the ethylacetate fraction of the extracts. Results showed that avicularin and quercetrin were active components of the extracts. These findings suggest that the M. quinquenervia leaf extracts can be applied to new cosmetics products as an effective antioxidant ingradient.

• Korean Journal of Food Science and Technology
• /
• v.42 no.4
• /
• pp.481-487
• /
• 2010

The nutritional components, antioxidant, and neuroprotective effects of water and a 50% methanol extract from litchi fruit pericarp were investigated. The most abundant mineral, amino acid, and fatty acid were K, proline, and palmitic acid, respectively. In addition, the total water phenolics and 50% methanol extracts were 8.02 and 12.28 mg/g, respectively. The DPPH, ABTS radical scavenging activities and ferric reducing antioxidant power of the water and 50% methanol extracts showed dose-dependent antioxidant activity. In a cell viability assay using MTT, almost all extracts showed a protective effect against $H_2O_2$-induced neurotoxicity, and lactate dehydrogenase leakage was also inhibited by the pericarp extracts. In particular, the 50% methanol extract showed a higher cell membrane protective effect than the water extract at the highest concentration. Consequently, these data suggest that litchi fruit pericarp can be utilized as an effective and safe functional food substances for natural antioxidants and may reduce the risk of neurodegenerative disorders.

• Journal of Life Science
• /
• v.18 no.6
• /
• pp.789-795
• /
• 2008

To study the possible use of probiotics in fish farming, we evaluated antagonism of antibacterial strain Bacillus amyloliquefaciens H41 against the fish pathogenic bacterium Vibrio anguillarum NCMB1. The purification of growth inhibition factor produced by B. amyloliquefaciens H41 was achieved by obtaining supernatant of this bacterium. The growth inhibition factor was purified to homogeneity by 70% ammonium sulfate precipitation, DEAE-sephadex A-50 ion exchange chromatography, sephadex G-200 gel filtration column chromatography, and sephadex G-50 gel filtration column chromatography with 40.8 fold of purification and 2.9% yield. The molecular weight of the purified growth inhibition factor was 48 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH and temperature for the growth inhibition factor were pH 7.5 and $30^{\circ}C$, respectively. The activity of growth inhibition factor was enhanced slightly by some metal ions, such as $Mg^{+2}$, $Mn^{+2}$, but was inhibited by the addition of $Co^{+2}$, $Hg^{+2}$, $Zn^{+2}$ and $Ag^{+2}$. NaCl stability of the growth inhibition factor was observed with 50% residual activity at 3% NaCl concentration. Toxicity test showed that the purified B. amyloliquefaciens H41 growth inhibition factor did not affect the live of Japanese flounder (Paralichthys olivaceus) and the effectiveness was 78% of residual lethality compared to commercial antibacterial agents.