• Proceedings of the Plant Resources Society of Korea Conference
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• 2022.09a
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• pp.31-31
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• 2022

Plant-derived antioxidants are used as a healthy diet and are known to inhibit various human diseases. In this study, we investigated free radical scavenging and antioxidant activity of extracts from three plants (Ammannia multiflora, Ammannia coccinea and Salix gracilistyla) with the most DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity from 196 plant extracts inhabiting along Nakdong River in Republic of Korea. The three extracts also have strong total antioxidant activity. Moreover, the extracts inhibited hydrogen peroxide (H₂O₂)-induced reactive oxygen species production and depolarized mitochondrial membrane potential in RAW264.7 macrophages. In zebrafish larvae, 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) fluorescent intensity, induced by H₂O₂, was markedly reduced by the extracts of A. multiflora, A. coccinea and S. gracilistyla. Meanwhile, the extracts were upregulated Nrf2 and HO-1 expression, and an HO-1 inhibitor reversed the extract-induced oxidative responses both in vivo and in vitro. The data suggest that the extracts of A. multiflora, A. coccinea, and S. gracilistyla exert potential free radical scavenging and antioxidant capacities both in vivo and in vitro by activating the Nrf2/HO-1 signaling pathway.

• Parasites, Hosts and Diseases
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• v.61 no.4
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• pp.388-396
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• 2023

Entamoeba histolytica is an enteric tissue-invasive protozoan parasite causing amoebic colitis and liver abscesses in humans. Amoebic contact with host cells activates intracellular signaling pathways that lead to host cell death via generation of caspase-3, calpain, Ca²⁺ elevation, and reactive oxygen species (ROS). We previously reported that various NADPH oxidases (NOXs) are responsible for ROS-dependent death of various host cells induced by amoeba. In the present study, we investigated the specific NOX isoform involved in ROS-dependent death of hepatocytes induced by amoebas. Co-incubation of hepatoma HepG2 cells with live amoebic trophozoites resulted in remarkably increased DNA fragmentation compared to cells incubated with medium alone. HepG2 cells that adhered to amoebic trophozoites showed strong dichlorodihydrofluorescein diacetate (DCF-DA) fluorescence, suggesting intracellular ROS accumulation within host cells stimulated by amoebic trophozoites. Pretreatment of HepG2 cells with the general NOX inhibitor DPI or NOX2-specific inhibitor GSK 2795039 reduced Entamoeba-induced ROS generation. Similarly, Entamoeba-induced LDH release from HepG2 cells was effectively inhibited by pretreatment with DPI or GSK 2795039. In NOX2-silenced HepG2 cells, Entamoeba-induced LDH release was also significantly inhibited compared with controls. Taken together, the results support an important role of NOX2-derived ROS in hepatocyte death induced by E. histolytica.

• Journal of the Korean Society of Tobacco Science
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• v.27 no.1 s.53
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• pp.31-39
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• 2005

This study was conducted to evaluate the characterization of the pyrolysis products of propylene glycol(PG) and glycerine alone and together with tobacco. The weight change of the samples during the pyrolysis was measured by a thermal analyzer(STD-2960). The pyrolysis products were determined by GC/MS after pyrolysis using a curie-point pyrolyzer(CPP, $220^{\circ}C,\;420^{\circ}C,\;650^{\circ}C,\;and\;920^{\circ}C$) and a double-shot pyrolyzer(DSP, $220^{\circ}C,\;420^{\circ}C,\;650^{\circ}C,\;and\;800^{\circ}C$), respectively. The pyrolysis products from tobacco with and without the addition of PG($2\%$) and glycerine($2\%$ were assayed for its pyrolytic behavior. The results showed that a dramatic change in weight of PG and glycerine was observed at $175^{\circ}C\;and\;249^{\circ}C$, respectively. PG and glycerine showed different patterns for their pyrolysis products according to the method of pyrolysis. Namely, the change rate in pyrolysis with DSP was much higher than that of CPP at above $650^{\circ}C$. The major pyrolysis products of PG were propene, acetaldehyde, propanal, and acetol; the major pyrolysis products of glycerine were 2-propenal, 2-propenol, acetol, and acetic acid. In the pyrolysis experiments of tobacco added PG and glycerine, the pyrolysis products of PG and glycerine weren't detected additionally, except for diethyleneglycol diacetate. From these results, it can be concluded that the PG and glycerine added to tobacco would not be expected to pyrolyse extensively during smoking.

• Korean Journal of Medicinal Crop Science
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• v.27 no.6
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• pp.365-375
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• 2019

Background: Oxidative stress plays an important role in neuro-degenerative disorders such as Alzheimer's disease. Oxidative stress is mediated by reactive oxygen species (ROS), which are implicated in the pathogenesis of numerous diseases, and account for the toxicity of a wide range of compounds. Methods and Results: In order to study the neuro-protective effect of the complex extracts of Aster scaber Thunberg (AS) and Chaenoleles sinensis Koehne (CSK) against hydrogen peroxide in PC12 cells, cell viability was evaluated by the MTT assay using tetrazole, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and the intracellular ROS levels were determined the by 2',7'-dichlorofluorescein diacetate (DCF-DA) assay. In order to examine the anti-amnesic effects of the complex extracts of AS and CSK, behavioral tests were performed on male ICR mice. The ameliorating effect of the complex extracts against Aβ_1-42-induced learning and memory impairment was analyzed by y-maze and passive avoidance tests. The AS and CSK extracts showed neuro-protective activity both in vitro and in vivo, and the neuro-protective effect of their 60 : 40 (AS : CSK) mixture was better than that of the other mixtures. Moreover, the complex extracts synergistically inhibited acetylcholinesterase activity and rapid peroxidation. Conclusions: A mixture of the AS and CSK extracts could be used to develop functional foods and serve as raw materials for the development of therapeutics against Alzheimer's disease.

• Korean Journal of Medicinal Crop Science
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• v.27 no.6
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• pp.419-426
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• 2019

Background: This study aimed to identify antioxidant compounds from the seeds of Dolichos lablab L. by bioassay-guided isolation and recrystallization. Methods and Results: The water layer of D. lablab L. seed extract inhibits intracellular reactive oxygen species (ROS) expressing the 2',7'-dichlorofluorescein diacetate (DCF-DA), Cu/Zn superoxide dismutase (SOD) and catalase genes, as determined by quantitative real-time PCR (qRT-PCR). Two compounds were purified from the water layer of the seeds of D. lablab L. using column chromatography and prep-high performance liquid chromatography (HPLC). Using nuclear magnetic resonance (NMR) and electrospray Ionization mass spectrometry (ESI-MS), their chemical structures were identified as 5-[(2-acetyl-2,3-dihydro-1H-indazol-1-yl)carbonyl]-4,5-dihydro-3H-furan-2-one (C₁₄H₁₄N₂O₄) and stachyose. Conclusions: Two active antioxidant compounds were purified from the seed extract of D. lablab L. seed extract and the structures of these compounds were identified as C₁₄H₁₄O₄N₂ and stachyose.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.5
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• pp.1833-1837
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• 2015

An aristolactam-type alkaloid, isolated from Orophea enterocarpa, is enterocarpam-III (10-amino-2,3,4,6-tetramethoxyphenanthrene-1-carboxylic acid lactam). It is cytotoxic to various human and murine cancer cell lines; however, the molecular mechanisms remain unclear. The aims of this study were to investigate cytotoxic effects on and mechanism (s) of human cancer cell death in human hepatocellular carcinoma HepG2 and human invasive breast cancer MDA-MB-231 cells compared to normal murine fibroblast NIH3T3 cells. Cell viability was determined by MTT assay to determine $IC_{10}$, $IC_{20}$ and $IC_{50}$ levels, reactive oxygen species (ROS) production with 2',7'-dichlorohydrofluorescein diacetate and the caspase-3, -8 and -9 activities using specific chromogenic (p-nitroaniline) tetrapeptide substrates, viz., DEVD-NA, IETD-NA and LEHD-NA and employing a microplate reader. Mitochondrial transmembrane potential (MTP) was measured by staining with 3, 3'-dihexyloxacarbocyanine iodide ($DiOC_6$) and using flow cytometry. The compound was cytotoxic to HepG2 and MDA-MB-231 cells with the $IC_{50}$ levels of $26.0{\pm}4.45$ and $51.3{\pm}2.05{\mu}M$, respectively. For murine normal fibroblast NIH3T3 cells, the $IC_{50}$ concentration was $81.3{\pm}10.1{\mu}M$. ROS production was reduced in a dose-response manner in HepG2 cells. The caspase-9 and -3 activities increased in a concentration-dependent manner, whereas caspase-8 activity did not alter, indicating the intrinsic pathway activation. Enterocarpam-III decreased the mitochondrial transmembrane potential (MTP) dose-dependently in HepG2 cells, suggesting that the compound induced HepG2 cell apoptosis via the mitochondrial pathway. In conclusion, enterocarpam-III inhibited HepG2 and MDA-MB-231 cell proliferation and induced human HepG2 cells to undergo apoptosis via the intrinsic (mitochondrial) pathway and induction of caspase-9 activity.

• Journal of Nutrition and Health
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• v.48 no.5
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• pp.451-456
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• 2015

Purpose: Neuronal apoptotic events induced by aging and hypoxic/ischemic conditions is an important risk factor in neurodegenerative diseases such as ischemia stroke and Alzheimer's disease. The peel of Citrus sunki Hort. ex Tanaka has long been used as a traditional medicine, based on multiple biological activities including anti-oxidant, anti-inflammation, and anti-obesity. In the current study, we examined the actions of fermented C. sunki peel extract against cobalt chloride ($CoCl_2$)-mediated hypoxic death in human neuroblastoma SH-SY5Y cells. Methods: Cell viability was measured by trypan blue exclusion. Expression of apoptosis related proteins and release of cytochrome c were detected by western blot. Production of intracellular reactive oxygen species (ROS) and apoptotic morphology were examined using 2',7'-dichlorofluorescin diacetate (DCF-DA) and 4',6-diamidino-2-phenylindole (DAPI) staining. Results: Exposure to $CoCl_2$, a well-known mimetic agent of hypoxic/ischemic condition, resulted in neuronal cell death via caspase-3 dependent pathway. Extract of fermented C. sunki peel significantly rescued the $CoCl_2$-induced neuronal toxicity with the cell viability and appearance of apoptotic morphology. Cytoprotection with fermented C. sunki peel extract was associated with a decrease in activities of caspase-3 and cleavage of poly (ADP ribose) polymerase (PARP). In addition, increase in the intracellular ROS and release of cytochrome c from mitochondria to the cytosol were inhibited by treatment with extract of fermented C. sunki peel. Conclusion: Based on these data, fermented C. sunki peel extract might have a protective effect against $CoCl_2$-induced neuronal injury partly through generation of ROS and effectors involved in mitochondrial mediated apoptosis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.6
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• pp.1475-1481
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• 2008

This study was to investigate the inhibitory effects of Mori Fructus on the generation of peroxynitrite ($ONOO^-$), nitric oxide (NO) and superoxide anion radical (${\cdot}O_2{^-}$) in the endothelial cells of rat vessels. The aim of this study was to investigate the $ONOO^-$, NO, ${\cdot}O_2{^-}$ scavenging and anti-inflammatory activitives of Mori Fructus. For this study, the fluorescent probes, namely dihydrorhodamine 123 (DHR 123), 4,5-diaminofluorescein (DAF-2) and 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) were used. Western blotting was performed using anti-NF-${\kappa}B$ (p50, p65), anti-COX-2, anti-iNOS antibodies, respectively. Mori Fructus prevented lipopolysaccharide (LPS)-induced cell death in YPEN cells. Mori Fructus inhibited the generation of $ONOO^-$, NO and ${\cdot}O_2{^-}$ in the LPS-treated cells. Mori Fructus inhibited the expression of COX-2 and iNOS genes by means of decreasing the NF-${\kappa}B$ activation. These results suggest Mori Fructus is effective on inhibiting the generation of $ONOO^-$, NO and ${\cdot}O_2{^-}$, and that therefore it might have a potential role as a treatment for the inflammatory process and inflammation-related diseases.

• Journal of the Korean Ceramic Society
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• v.39 no.3
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• pp.299-307
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• 2002

In this proposed study, a new emerging shape forming technique Direct Coagulation Casting(DCC) which enables to fabricate complex-shaped ceramic parts has been investigated using colloid surface chemistry. Various process variables affected by dispersant, coagulation agent and sintering additives, have been evaluated in order to achieve highly concentrated stabilized silicon nitride suspensions. A high solid loading of 51 vol% in the dispersed silicon nitride suspension was prepared with 1.0wt% Tetraethylammonium Hydroxide (TEAH), which obtained a stable silicon nitride suspension with sintering additives $(Al_2O_3\;and\;Y_2O_3)$ in alkaline regions. The addition of hydroxyaluminium diacetate into the suspension, which decomposed at elevated temperatures, led to coagulate of a silicon nitride suspension. In a basic medium, aluminum ions precipitated to aluminum hydroxide $(Al(OH)_3)$, leading to decreased $OH^-$ concentration and, thus, coagulated suspension.

• Journal of Electrochemical Science and Technology
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• v.6 no.3
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• pp.106-110
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• 2015

This study investigates Cu(II) complexes of cyclam, propylene cross-bridged cyclam (PCB-cyclam), and propylene cross-bridged cyclam diacetate (PCB-TE2A) as homogeneous electrocatalysts for CO₂ reduction in comparison with Ni(II)-cyclam. It is found that Cu(II)-cyclam can catalyze CO₂ reduction at the potential close to its thermodynamic value (0.75 V vs. Ag/AgCl) in tris-HCl buffer (pH 8.45) on a glassy carbon electrode. Cu(II)-cyclam, however, suffers from severe demetalation due to the insufficient stability of Cu(I)-cyclam. Cu(II)-PCB-cyclam and Cu(II)-PCB-TE2A are revealed to exhibit much less demetalation behavior, but poor CO₂ reduction activities as well. The inferior electrocatalytic ability of Cu(II)-PCB-cyclam is ascribed to its redox potential that is too high for CO₂ reduction, and that of Cu(II)-PCB-TE2A to the steric hindrance preventing facile contact with CO₂ molecules. This study suggests that in addition to the redox potential and chemical stability, the stereochemical aspect has to be considered in designing efficient electrocatalysts for CO₂ reduction.