• Environmental Engineering Research
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• v.14 no.1
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• pp.19-25
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• 2009

In this study, total bacteria, enteric members of the $\gamma$-proteobacteria, and microbial communities in seawater were analyzed as indirect indicators for quantifying biofouling. Biomass in seawater can significantly affect feed water pretreatment and membrane biofouling of reverse osmosis desalination processes. The purpose of this paper is to investigate microbiological quantity and quality of seawater at the potential intake of a desalination plant. For this analysis, the total direct cell count (TDC) using 4'-6-diamidino-2-phenylindole (DAPI)-staining and DNA-based real-time PCR were used to quantify the total bacteria and relative content of enteric members of $\gamma$-proteobacteria in seawater, respectively. In addition, microbial communities were examined using 16S rRNA gene cloning and bacterial isolation to identify the most abundant bacteria for a further biofouling study. The experimental results of this study identified about $10^6$ cells/mL of (total) bacteria, $10^5$ 16S rRNA gene copies/mL of enteric $\gamma$-proteobacteria, and the presence of more than 20 groups of bacteria.

• International Journal of Oral Biology
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• v.45 no.3
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• pp.99-106
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• 2020

Ficus carica L. (fig) is one of the first cultivated crops and is as old as humans. This plant has been extensively used as a traditional medicine for treating diseases, such as cough, indigestion, nutritional anemia, and tuberculosis. However, the physiological activity of fig leaves on oral cancer is as yet unknown. In this study, we investigated the anticancer effect of methanol extracts of Ficus carica (MeFC) and the mechanism of cell death in human FaDu hypopharyngeal squamous carcinoma cells. MeFC decreased the viability of oral cancer (FaDu) cells but did not affect the viability of normal (L929) cells, as determined by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay and Live and Dead assay. In addition, MeFC induced apoptosis through the proteolytic cleavage of procaspase-3, -9, poly (ADP-ribose) polymerase (PARP), downregulation of Bcl-2, and upregulation of Bax, as determined by 4′,6-diamidino-2-phenylindole dihydrochloride staining and western blot analysis. Moreover, a concentration of MeFC without cytotoxicity (0.25 mg/mL) significantly suppressed colony formation, a hallmark of cancer development, and completely inhibited the colony formation at 1 mg/mL. Collectively, these results suggest that MeFC exhibits a potent anticancer effect by suppressing the growth of oral cancer cells and colony formation via caspase- and mitochondrial-dependent apoptotic pathways in FaDu human hypopharyngeal squamous carcinoma cells. Therefore, the methanol extract of Ficus carcica leaves provide a natural chemotherapeutic drug for human oral cancer.

• Journal of Korean Medicine Rehabilitation
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• v.24 no.2
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• pp.41-50
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• 2014

Objectives The purpose of this study was to evaluate the effects of Clematidis radix extract on $CoCl_2$-induced apoptosis in SH-SY5Y human neuroblastoma cells. Methods In order to investigate the protective effect of Clematidis radix on $CoCl_2$-induced cytotoxicity in neuronal cells, MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, DAPI(4,6-diamidino-2-phenylindoleI) staining, TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling) assay, DNA fragmentation assay and western blotting were performed on SH-SY5Y human neuroblastoma cells. Results Cells treated with $CoCl_2$ exhibited several apoptotic features, while cells pre-treated with Clematidis radix prior to $CoCl_2$ exposure showed a decrease in the occurrence of apoptotic features. $CoCl_2$ increased HIF-$1{\alpha}$ expression, in contrast, Clematidis radix treatment decreased $CoCl_2$-induced HIF-$1{\alpha}$ expression. Pre-treatment with the extract of Clematidis radix suppressed Bax, cytochrome c, and caspase-3 expressions, and also increased Bcl-2 expression in SH-SY5Y human neuroblastoma cells. Conclusions These results suggest that Clematidis radix may exert a protective effect on $CoCl_2$-induced apoptosis in SH-SY5Y human neuroblastoma cells.

• Bulletin of the Korean Chemical Society
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• v.28 no.12
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• pp.2203-2208
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• 2007

Three possible binding modes of cationic meso-tetrakis(N-methylpyridinium-4-yl)porphyrin (TMPyP) to d[(GCATATATGC)2] duplex were investigated by the molecular dynamics (MD) simulation. Among the three binding modes namely, “along the groove”, “across the groove” and “face on the groove”, the “across the groove” model exhibited the largest negative binding free energy and the DNA backbone remained as the B form. In this model, the molecular plain of the TMPyP tilts 45o with respect to the DNA helix axis and is largely exposed to the solvent. TMPyP was stabilized mainly by the interaction between the positively charged neighboring pyridinium moieties of TMPyP and negatively charged phosphate groups of DNA. The result obtained in this work by MD and the report (Jin, B. et al., J. Am. Chem. Soc. 2005, 127, 2417.) that the spectral properties of poly[d(A-T)2] bound TMPyP in the presence and absence of the minor groove binding drug 4',6- diamidino-2-phenylindole are similar, we propose that TMPyP bind across the minor groove of the AT rich- DNA.

• Preventive Nutrition and Food Science
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• v.12 no.2
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• pp.79-83
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• 2007

The effects of young radish (YR, yeolmu in Korean) on the induction of apoptosis were examined in AGS human gastric adenocarcinoma cells. The young radish kimchi (YRK) were made of YR cultivated in the soil without (Control YR kimchi: C-YRK) and with 1,818 g/m$^{3}$ sulfur (Sulfur YR kimchi: S-YRK), respectively. Methanol extracts from S-YRK exhibited higher inhibitory effect on the growth of AGS human gastric adenocarcinoma cells in a time dependent-manner compared to C-YRK at the same concentration. 4,6-diamidino-2-phenylindole (DAPI) staining showed that S-YRK induced apoptosis accompanied by the increased Bax but decreased Bcl-2 in mRNA expression. Moreover, S-YRK decreased the levels of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) mRNA expressions. The results suggested that S-YRK cultivated in the presence of sulfur elicited stronger anticancer activity than C-YRK in vitro. Dietary intakes of S-YRK may be beneficial to decrease the risk of cancer.

• The Journal of Korean Medicine
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• v.32 no.1
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• pp.109-120
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• 2011

Objectives: The aim of this investigation was to evaluate the efficacy of KHchunggan-tang aqueous extract on the experimental nonalcoholic fatty liver disease(NAFLD) induced by palmitate. Materials and Methods: To generate a cellular model of NAFLD, we used HepG2 cells, a human hepatoma cell line, treated with 0.5 mM palmitate. By this cellular model, effects of KHchunggan-tang aqueous extract were evaluated. Intracellular lipid accumulation, free radical formation, and apoptosis were detected by Nile red staining, 2',7'-dichloroflourescin diacetate(H2DCF-DA), and 4',6-diamidino-2-phenylindole(DAPI)/propidium iodide(PI) staining, respectively. Some proteins related with NAFLD were determined by western blot. Results: Typical pathological features of NAFLD occurred in the cellular model. Palmitate increased the levels of intracellular lipid vacuoles, decreased cell viability, and increased apoptosis. Palmitate increased free radical formation and lipid peroxidation, too. However, KHchunggan-tang aqueous extract reduced palmitate-induced pathologic features, i.e. steatosis, free radical formation, and apoptosis. In addition, KHchunggan-tang aqueous extract suppressed palmitate-activated c-Jun N-terminal kinase(JNK) signaling, and SP600125, a JNK inhibitor, significantly reversed the palmitate-induced pathologic changes as KHchunggan-tang aqueous extract. It means that the signaling pathway other than JNK can be involved in the KHchunggan-tang mediated cellular protection of palmitate-treated Hep G2 cells. Conclusions: These results suggest that KHchunggan-tang aqueous extract has hepatoprotective effects on NAFLD with combined properties in cellular steatosis, ROS production, and cytoprotection, and thus may have valuable clinical applications for treatment of this chronic liver disease.

• Journal of Photoscience
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• v.4 no.3
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• pp.97-103
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• 1997

The effect of temperature on the fluorescence quenching of 2-(4-Methoxyphenyl)-5-(1-naphthyl)-1,3,4-oxadiazole (MPNO1), 2-(4-Methoxyphenyl)-5-(2-naphthyl)-1,3,4-oxadiazote(MPNO2), by aniline, and 2-Phenylindole (2-PI) by CCk, in toluene by steady state method and in benzene by time-resolved method have been carried out in the temperature range 30 - 70$\circ$C. The Stem-Volmer (S-V) plots, I$_0$/I against quencher concentration [Q] at different temperanares show positive deviations. The fluorescence lifetimes determined at different temperatures show no systematic variations and the variations being within the experimental error, the average values of lifetimes $ $\tau$ (t) are taken for further calculations. Rate constants such as Stem-Volmer quenching constants K$_sv}$, quenching rate parameters k$_q$ and k'$_q$, static quenching constant V and kinetic distance r are determined using the modified Stem-Volmer equation and sphere of action static quenching model. In order to see whether the reactions are diffusion limited, equations k$_q$ = e$^{-Eq/RT}$ and k'$_q$ = e$^{-Eq/RT}$ are used to determine the values of E$_q$ and E'$_q$, the activation energies for collisional quenching and the values of E$_q$ are 14.53. 17.28 and 16.20 kJ mole$^{-1}$ for MPNO1, MPNO2 and 2-PI respectively and the values of E'$_q$ are 14.62 and 17.73 for MPNO1 and MPNO2 respectively. From the magnitudes of various quantities it has been concluded that the reactions are diffusion limited and the observed positive deviations in the S-V plot are due to static and dynamic quenching.

• Journal of Life Science
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• v.14 no.2
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• pp.255-262
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• 2004

We applied nuclear DNA content stained with 4'-6'-diamidino-2-phenylindole (DAPI) and total protein concentration to predict the existence of Cochlodinium polykrikoides before huge blooms occurred, based on a short-term survey at sites in the South Sea. Fluctuations in environmental conditions and nutrients (nitrate, nitrite, and phosphate) were of a similar range, regardless of sampling sites or early and middle field observations. However, C. polykrikoides abundance was significantly different depending on the station, with a higher cell density of 34, 62, and 57 cells L$^{-1}$ at Stn C2, C5, and C6, respectively than what was found in early August, 2000. In mid August, 2000, the highest cell density of 547 cells L$^{-1}$ at Stn C3 was observed. The relationship between C. polykrikoides abundance, DAPI-stained DNA content, and total protein concentration was a positive correlation coefficient, in particular a higher positive correlation was exposed to even a smaller abundance of C. polykrikoides. These results suggest that DNA stained by DAPI and total protein concentration could play an important index in easily predicting the presence of C. polykrikoides before blooms.

• Journal of Korean Medicine for Obesity Research
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• v.15 no.2
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• pp.93-103
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• 2015

Objectives: It was investigated the cytoprotective efficacies of isorhamnetin, a flavonoid originally derived from Hippophae rhamnoides L., against oxidative stress-induced apoptosis in C2C12 myoblasts. Methods: The effects of isorhamnetin on cell growth, apoptosis and reactive oxygen species (ROS) generation were evaluated by trypan blue dye exclusion assay, 4',6-diamidino-2-phenylindole staining and flow cytometry. The levels of apoptosis-regulatory and nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway-related proteins, and caspase activities (caspase-3 and -9) were determined by Western blot analysis and colorimetric assay, respectively. Results: Our results revealed that treatment with isorhamnetin prior to hydrogen peroxide ($H_2O_2$) exposure significantly increased the C2C12 cell viability and, indicating that the exposure of C2C12 cells to isorhamnetin conferred a protective effect against oxidative stress. Isorhamnetin also effectively attenuated $H_2O_2$-induced apoptosis and ROS generation, which was associated with the restoration of the upregulation of Bax and downregulation of Bcl-2 induced by $H_2O_2$. In addition, $H_2O_2$ enhanced the activation of caspase-9 and -3, and degradation of poly (ADP-ribose)-polymerase, a typical substrate protein of activated caspase-3; however, these events were almost totally reversed by pretreatment with isorhamnetin. Moreover, isorhamnetin increased the levels of heme oxygenase-1, a potent antioxidant enzyme, associated with the induction of Nrf2. Conclusions: Our data indicated that isorhamnetin may potentially serve as an agent for the treatment and prevention of muscle disorders caused by oxidative stress.

• The Journal of Korean Medicine
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• v.28 no.1 s.69
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• pp.198-210
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• 2007

Objectives : Nitric oxide(NO) is a reactive free radical and a messenger molecule in many physiological functions. However, excessive release NO of induces neurotoxicity. We investigated whether a mixture of red ginseng and paeonia radix prossesses a protective effect against sodium nitroprusside(SNP)-induced apoptosis in the human neuroblastoma cell line SK-N-MC. Methods : We performed 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, 4,6-diamidino-2-phenylindole(DAPD) staining, terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL)assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction(RT-PCR), Western blot analysis, and caspase-3 enzyme activity assay in SK-N-HFC cells. Result : MTT assay showed that SNP treatment significantly reduced the viabilities of cells and that pre-treatment with the red ginseng and paeonia radix mixture alleviated SNP-induced cytotoxicity. The cells treated with SNP exhibited several apoptotic features, while those pre-treated fir 1 h with the mixture of red ginseng and paeonia radix 1 h prior to SNP expose showed reduced apoptotic features. In addition, the cells pre-treated with the red ginseng and paeonia radix mixture for 1 h prior to SNP expose increased bel-2 expressions, decreased Bax expressions, and decreased caspase-3 enzyme activity. Conclusions : These results show that the red ginseng and paeonia radix mixture exerts a protective effect against SNP-induced apoptosis in SK-N-MC cells.