• Microbiology and Biotechnology Letters
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• v.48 no.1
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• pp.79-89
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• 2020

This study was performed to evaluate the anti-inflammatory activities of 70% ethanol extract from Ambrosia trifida L. (AT). The electron donating ability and ABTS⁺ radical scavenging ability of extract from AT was shown to be 84.1% and 92.5% at 1,000 ㎍/ml concentration. The astringent effect of extract from AT was shown to be 94.7% at 1,000 ㎍/ml. The anti- inflammatory activities of extract of AT were investigated using RAW 264.7 cells induced by lipopolysaccharide (LPS). The cell toxicity effect of AT extract on RAW 264.7 performed MTT assay. As a result of the measured cell toxicity effect, 90% or more was shown with cell viability at a 500 ㎍/ml concentration. In nitric oxide synthesis inhibition effect, it was shown that extract from AT concentration dependent inhibited nitric oxide production. The protein expression inhibitory effect of AT extract was measured by western blot at 25, 50, and 100 ㎍/ml concentration and the β-actin used as a positive control. Consequently, the inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 protein expression inhibitory effect was decreased by 8.6%, 25.1% at 100 ㎍/ml concentration. The phosphorylation of extracellular signal-regulated kinase 1/2, p38, c-Jun NH₂-terminal kinase and Iκ-Bα protein expression inhibitory effect was a decreased dependent concentration. The mRNA expression inhibitory effect was measured by reverse transcription - polymerase chain reaction at 25, 50, and 100 ㎍/ml concentration and the glyceraldehyde-3-phosphate dehydrogenase used as a positive control. Consequently, the iNOS, COX-2, interleukin (IL)-1β, IL-6 and tumor necrosis factor-α mRNA expression inhibition effect was a decreased dependent concentration in an LPS-activated macrophage. In conclusion, AT extract may have some effects on inflammatory factors as potential anti-inflammatory agents and natural substance for cosmetics.

• Journal of fish pathology
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• v.28 no.2
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• pp.63-69
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• 2015

The biological characteristics of Bacillus sp.CPB-St as a probiotic strain to control fish streptococcosis was determined. Based on 16S rRNA sequencing, Bacillus sp.CPB-St was identified as Bacillus pumilus and named B. pumilus CPB-St (Abbreviated as CPB-St). Growth inhibitory activity of CPB-St against Streptococcus spp. was examined at three different incubation temperatures ($20^{\circ}C$, $25^{\circ}C$, and $30^{\circ}C$) and three culture media (NA, TSA, and BHIA) based on the diameter of inhibition zone. Its activity (inhibition zone of 11~29 mm) at $20^{\circ}C$ was higher than that (12~21 mm) at $30^{\circ}C$. Its activity (29 mm) in NA media was the same as that (29 mm) in TSA media. However, it was higher than that (22 mm) in BHIA media. The inhibitory activity of CPB-St against Streptococcus spp. was high at pH7. However, its activity was the same at salinity of 0.5% to 3%. CPB-St showed maximum growth after incubation at $25^{\circ}C$ for 48 h. To use CPB-St as probiotics, settlement studies in fish intestine and its efficacy through feeding are needed. CPB-St was highly resistant to gastric juice at pH4 and flounder's bile salt as well as deoxycholic acid at $300{\mu}g/ml$. CPB-St showed optimal viability in 1% NaCl. It showed similar growth in 0% to 7% NaCl. CPB-St could tolerate $-20^{\circ}C$ and $-70^{\circ}C$ for 45 min. There was no difference in the growth of the strain between room temperature and $4^{\circ}C$. Fish diet supplemented with CPB-St could be stored at low temperature without cell loss. Therefore, CPB-St might be used as probiotics to control streptococcosis of fish.

• Korean Journal of Food Science and Technology
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• v.50 no.4
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• pp.383-390
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• 2018

Most neurodegenerative diseases are known to be influenced by oxidative stress. We investigated the anti-oxidative activity of the concentrate of fermented wild ginseng root culture (HLJG0701) containing ginsenosides Rg5 and Rk1. HLJG0701 showed effective DPPH and ABTS radical scavenging ability ($IC_{50}$: 16- and 4-fold dilution, respectively) and was inhibited dose-dependently by the $FeSO_4$-induced lipid peroxidation group (8- and 4-fold dilution: 2.3 and 1.5 nM, respectively). In MTT and LDH assays, 8-, 16-, 32- and 64-fold diluted HLJG0701 significantly increased cell viability by 70, 53, 35, and 26%, respectively. LDH released by HLJG0701 was reduced 1.3-fold with 8-fold diluted HLJG0701 compared to the $H_2O_2$-treated control. In addition, the inhibitory effect of HLJG0701 on oxidative stress in PC12 cells was confirmed by DCF-DA analysis (16-, 4-fold diluted HLJG0701: 50 and 68% ROS inhibition, respectively), TBARS (16- and 4-fold diluted HLJG0701: 50.7 and 46.5% inhibition, respectively), GPx (16- and 4-fold diluted HLJG0701: 133.3 and 227.3% release, respectively), and SOD analysis (16- and 4-fold diluted HLJG0701: 118.2 and 218.2% release, respectively). These results suggested that HLJG0701 protects neuronal cells by its anti-oxidative effects and hence can be a potential preventive material against neurodegenerative diseases.

• The Korean Journal of Nuclear Medicine
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• v.37 no.5
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• pp.308-316
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• 2003

Purpose: The green tea polyphenol (GTPP) has been known to exert antioxidant activity as a radical scavenger as well as cancer preventive and cancer growth inhibition effect. The aim of this study was to identify whether GTPP not only potentiate the growth inhibition effect in ${\gamma}-irradiated$ human cancer cell but also exert protection action for irradiated human normal cell. Materials and Methods: GTPP (80% catechin including >45% EGCG) added in the HL60, human leukemia, and NC37, human lymphoblast, before irradiation. After establishing the amount of GTPP and the dose of radiation, the cells were treated with the GTPP for 6 hours and irradiated with the determined doses. Results: Viability when $10{\mu}g/ml$ GTPP added before ${\gamma}-irradiation$ with 1 Gy to NC37 cells was not different in comparison with control but it when was irradiated with 3 Gy significantly different (1 Gy;P=0.126, 3 Gy;P=0.010). $20{\mu}g/ml$ GTPP did not show significant difference in both NC37 cells irradiated with 1 Gy and 3 Gy (1 Gy;P=0.946, 3 Gy;P=0.096). Viabilities were significantly decreased with concentration of additional GTPP in HL60 with 1 or 3 Gy (1 Gy $69.0{\pm}1.7%\;vs\;42.4{\pm}1.3%,\;3\;Gy;\;66.9{\pm}3.9%\;vs\;44.2{\pm}1.6%$). Conclusion: In vitro study, we certified that when the cells were irradiated with dose below 3 Gy, GTPP provide not only anticancerous effect against cancer cells but also radioprotective effect in normal cells simultaneously. Theses results suggest the possibility that consumption of green tea could give the radioprotective effect and maximize the effect on internal radiation such as radioiodine therapy concomitantly.

• Applied Biological Chemistry
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• v.51 no.2
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• pp.142-147
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• 2008

Nuruk is the Korean traditional Koji that contains various microorganisms and has been used to make the traditional fermented foods including alcoholic beverages. Rhizopus oryzae KSD-815 was isolated from the alcohol-fermenting Nuruk used for manufacturing traditional alcohol. In this study, the authors reported the isolation and identification of four lipids from the Nuruk (Rhizopus oryzae KSD-815) that inoculated wheat with Rhizopus oryzae KSD-815. The dried and powdered Nuruk (Rhizopus oryzae KSD-815) were extracted three times at room temperature with 80% aqueous MeOH. The extracts were partitioned with EtOAc, n-BuOH, and water, successively. The EtOAc extract was suspended in 80% MeOH and partitioned repeatedly with n-hexane. From the n-hexane fraction, four lipids were isolated through the repeated silica gel and ODS column chromatographies. According to the results of physico-chemical data including NMR, GC and MS, the chemical structures of the compounds were determined as linolenic acid methyl ester (1), palmitic acid methyl ester (2), linoleic acid (3), palmitic acid (4). Cytotoxicity was evaluated in huamn breast cancer cells, MDA-MB-231 and human hepatocarcinoma, SK-HEP-1 cells using MTT assay. Exposure of compounds 1 and 3 led to a dose-dependent inhibition of cell viability in both cancer cell lines. In addition, treatment of RAW264.7 cells with compound 3 caused inhibition of lipopolysaccharide/interferon-${\gamma}$-induced nitric oxide production.

• Tuberculosis and Respiratory Diseases
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• v.65 no.6
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• pp.476-486
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• 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 Oral Medicine and Pain
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• v.36 no.1
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• pp.11-20
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• 2011

Valproic acid (VPA) is a well-known anticonvulsive agent and has been used in the treatment of epilepsy for almost 30 years. VPA emerged in 1997 as an antineoplastic agent. And it is known that antitmor activity of VPA is associated with its targeted at histone deacetylases. 17AAG, Inhibition of HSP90 leads to the proteasome degradation of the HSP90 client proteins, such as Akt, Raf/Ras, Erk, VEGF, cyclin D and p53, and causes potent antitumor activity. It is reported that 17AAG-induced HSP90 inhibition results in prevention of cell proliferation and induction of apoptosis in several types of cancer. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with the histone deacetylases inhibitor, VPA and the HSP90 inhibitor, 17AAG on human osteosarcoma (HOS) cells. Cell viability was evaluated by trypan-blue exclusion. Induction and augmentation of apoptosis were confirmed by Hoechst staining, flow cytometry (DNA hypoploidy and MMP change), Westen blot analysis and immunofluorescent staining. In this study, HOS cells co-treated with VPA and 17AAG showed several lines of apoptotic manifestation such as nuclear condensations, the reduction of MMP, the decrease of DNA content, the release of cytochrome c into cytosol, the translocation of AIF onto nuclei, and activation of caspase-3, caspase-7 and PARP whereas each single treated HOS cells did not. Although the single treatment of 1 mM VPA or 0.5 ${\mu}M$ 17AAG for 48 h did not induce apoptosis, the co-treatment with them induced prominently apoptosis. Therefore our data in this study provide the possibility that combination therapy with VPA and 17AAG could be considered as a novel therapeutic strategy for human osteosarcoma.

• Journal of Life Science
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• v.26 no.12
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• pp.1431-1437
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• 2016

Sorghum bicolor L. is one of the important minor cereals in Asia, Africa, and the central United States, and it is considered a rich source of polyphenols, flavonoids, and dietary fiber. However, there is a lack of data on the anti-cancer activity of Sorghum in prostate cancer cells and immune activity in macrophages. This study aims to investigate the potential effects of an ethanol extract of S. bicolor L. (SE) on inducing apoptosis in RC-58T/h/SA#4 cells and immunomodulatory activity in RAW 264.7 cells. SE significantly inhibited the viability of RC-58T/h/SA#4 primary prostate cancer cells in a dose-dependent manner. The morphology of RC-58T/h/SA#4 cells treated with SE was shrunken and involved the formation of an apoptotic body and nuclear condensation. In addition, SE markedly activated caspase-8, -9, and -3; increased the protein levels of Bax, p53, cleaved PARP, and cytosolic cytochrome c; and decreased Bcl-2 protein expression. Furthermore, the inhibition of caspases in RC-58T/h/SA#4 cells with z-VAD-fmk attenuated SE-induced cell growth inhibition. The production of nitric oxide (NO) was also elevated by SE treatment, as revealed by immune response parameters. These results suggest that SE inhibits growth and induces apoptosis in primary human prostate cancer cells in a caspase-dependent manner, and it modulates the immune functions in macrophages. Therefore, Sorghum bicolor L. may be used as a functional food to prevent prostate cancer and enhance immune activity.

• Journal of the East Asian Society of Dietary Life
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• v.19 no.3
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• pp.384-394
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• 2009

This study was performed to determine the antioxidative and anticancer effects of extracts from Adenophora remotiflora leaves. The antioxidative effects of the extracts were measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH)-radical scavenging activity and hemoglobin-induced linoleic acid oxidative inhibition assays. The results indicated that the extracts had stronger effects than the synthetic antioxidant BHT at the same concentration. The $SC_{50}$ values (50% radical scavenging effect on $1{\times}10^{-4}$ M DPPH) of the methanol fraction, water extract, and BHT were 47.5 ${\mu}g$/mL, 74.6 ${\mu}g$/mL and 102.2 ${\mu}g$/mL, respectively. In addition the $IC_{50}$ values (hemoglobin-induced linoleic acid oxidation inhibition) of the methanol fraction, water extract, and BHT were 120.8 ${\mu}g$/mL, 135.6 ${\mu}g$/mL, and 150.2 ${\mu}g$/mL, respectively. This research also assessed decreases in the survival of BNLcl2 cells (normal liver cells) by solvent fractions of the A. remotiflora leaf extracts at various concentrations (1, 5, 10, 25, 50, 100, 250, 500, 1,000, 2,000 ${\mu}g$/mL). The water extract did not decrease survival at any of the concentrations when compared to the control group. The hexane, ethyl acetate, and methanol fractions decreased survival as compared to the control group by inducing cell toxicity at a concentration of 1,000 ${\mu}g$/mL and above. Therefore, an anticancer activity experiment was conducted using concentrations below 500 ${\mu}g$/mL. At 500 ${\mu}g$/mL, the methanol fraction decreased A549 cell (human lung carcinoma cells) survival by 46% as compared to the control group, presenting the greatest effect against cell survival. All extracts showed greater anticancer activity in Hep G2 cells (human liver carcinoma cells) as compared to the A549 cells. For the Hep G2 cells, the methanol extract decreased survival by 28% as compared to the control group at the concentration of 500 ${\mu}g$/mL, thus restraining lung cancer cell growth.

• Journal of Life Science
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• v.23 no.2
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• pp.213-221
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• 2013

Bibimbab (mixed rice) is a traditional Korean one-dish meal. This study was carried out to investigate the anti-inflammatory and antioxidative effects of raw and seasoned ingredients used in Bibimbab (Cucurbita moschata P., Platycodon grandiflorum A., Vigna radiata L., Porphyra yezonensis udea, Allium ampeloprasum L., Pterdium aguilinum, Raphanus sativus). Human mast cells (HMC-1) were pretreated with 70% ethanol extracts of Bibimbab and further cultured for an appropriate time after the addition of phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187. Cell viability was determined by an MTT assay. None of the ingredients showed cytotoxic effects at a concentration of 1.0 ${\mu}g/ml$. Anti-inflammatory effects were analyzed at 0.01, 0.1, and 1.0 mg/ml concentrations of the raw, seasoned ingredients of PMA. A23187 stimulated HMC-1. Among the various ingredients, seasoned A. ampeloprasum L. extract showed the highest inhibition of TNF-${\alpha}$ and IL-6 secretion (90% and 93%, respectively) at a concentration of 1.0 mg/ml. The R. sativus extract showed the highest inhibition (85%) of IL-8 secretion. DPPH analysis of the antioxidation properties of the ingredients showed that raw and seasoned A. ampeloprasum extracts exhibited the highest DPPH free radical scavenging activity (67.50 and 73.65%, respectively). These results suggest that seasoned ingredients used in Bibimbab have lower anti-inflammatory effects in relation to TNF-${\alpha}$ and IL-6 secretion than raw ingredients in PMA- and A23187-treated HMC-1. In addition, the seasoned ingredients showed a tendency to increase antioxidative activity. Therefore, the ingredients used in Bibimbab have potential as anti-inflammatory and antioxidation agents.