• Food Science and Preservation
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• v.19 no.3
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• pp.455-461
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• 2012

Obesity, a strong risk factor for the development of chronic diseases, is characterized by an increase in the number and size of adipocytes differentiated from precursor cells, preadipocytes. Recent research suggests that increased reactive oxygen species (ROS) production in 3T3-L1 adipocyte facilitates adipocyte differentiation and fat accumulation. This study was to investigate whether reduced ROS production by Sargassum micracanthum extract (SME) could protect the development of obesity through inhibition of adipogenesis. 3T3-L1 preadipocytes were treated SME for up to 8 days following standard induction of differentiation. The extent of differentiation reflected by amount of lipid accumulation and ROS production was determined by Oil red O staining and nitroblue tetrazolium (NBT) assay. Treatment of SME significantly inhibited ROS production and adipocyte differentiation that is depend on down regulation of NADPH oxidase 4 (NOX4), a major ROS generator, and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein alpha ($C/EBP{\alpha}$), a key adipogenic transcription factor. These results indicate that SME can inhibit adipogenesis through a reduced ROS level that involves down-regulation of NOX4 expression or via modulation of adipogenic transcription factor.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.392-402
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• 2017

Background: Previously, we reported that Korean Red Ginseng inhibited liver fibrosis in mice and reduced the expressions of fibrogenic genes in hepatic stellate cells (HSCs). The present study was undertaken to identify the major ginsenoside responsible for reducing the numbers of HSCs and the underlying mechanism involved. Methods: Using LX-2 cells (a human immortalized HSC line) and primary activated HSCs, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assays were conducted to examine the cytotoxic effects of ginsenosides. $H_2O_2$ productions, glutathione contents, lactate dehydrogenase activities, mitochondrial membrane permeabilities, apoptotic cell subpopulations, caspase-3/-7 activities, transferase dUTP nick end labeling (TUNEL) staining, and immunoblot analysis were performed to elucidate the molecular mechanism responsible for ginsenoside-mediated cytotoxicity. Involvement of the AMP-activated protein kinase (AMPK)-related signaling pathway was examined using a chemical inhibitor and small interfering RNA (siRNA) transfection. Results and conclusion: Of the 11 ginsenosides tested, 20S-protopanaxadiol (PPD) showed the most potent cytotoxic activity in both LX-2 cells and primary activated HSCs. Oxidative stress-mediated apoptosis induced by 20S-PPD was blocked by N-acetyl-$\text\tiny L$-cysteine pretreatment. In addition, 20S-PPD concentration-dependently increased the phosphorylation of AMPK, and compound C prevented 20S-PPD-induced cytotoxicity and mitochondrial dysfunction. Moreover, 20S-PPD increased the phosphorylation of liver kinase B1 (LKB1), an upstream kinase of AMPK. Likewise, transfection of LX-2 cells with LKB1 siRNA reduced the cytotoxic effect of 20S-PPD. Thus, 20S-PPD appears to induce HSC apoptosis by activating LKB1-AMPK and to be a therapeutic candidate for the prevention or treatment of liver fibrosis.

• Journal of Life Science
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• v.26 no.8
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• pp.928-935
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• 2016

Hypsizygus marmoreus (white cultivar), also called white beech mushrooms, are edible mushrooms commercially cultivated in Korea and Japan. This study was carried out to evaluate the antioxidant properties of H. marmoreus. H. marmoreus fruit bodies were divided into pileus and stipe. The pileus and stipe were extracted into water and 80% ethanol and their antioxidant activities were analyzed. The total polyphenol content was highest in the water extract (pileus 1137.39±0.38 mg of GAE (gallic acid equivalents)/100 g, stipe 700.86±0.06 mg of GAE/100 g) compared to the ethanol extract (pileus 923.47±0.18 mg of GAE/ 100 g, stipe 324.05±0.03 mg of GAE/100 g). Ethanol extracts from pileus showed better scavenging ability on DPPH (47.32±0.23% at 10 mg/ml) and ABTS (57.33±0.10% at 10 mg/ml) than the stipe and water extract groups. Water extract from pileus were more effective in reducing power and ORAC (oxygen radical absorbance capacity) value than stipe and ethanol extract. The toxicity of water and ethanol extracts was investigated using WST-1 (Water Soluble Tetrazolium salt) assay on the mouse macrophage cell line RAW 264.7. Overall, total polyphenol content and antioxidant activities of extracts from H. marmoreus increased in a dose dependent manner while pileus was showed better total polyphenol content and antioxidant activities than stipe.

• Korean Journal of Microbiology
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• v.5 no.2
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• pp.69-78
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• 1967

Kim, Jong Hyup, (Div. of Biology, Atomic Energy Research Institute.) Studies on the Cellular Metabolism in Microorganisms as influenced by Gamma-irradiation(II). On respiration rate and dehydrogenase activity of yeast cells irradiated by gamma ray from cobalt-60. 1. Oxygen uptake rate of the gamma irraiated yeast cells had been measured with Warburg's manometer, and the $O_{2}$-uptake was compared with those of normal cells. The rate of endogetious respiration increases in its $O_2$-uptake at 150, 000 rentgen dose, and at higher rentoen doses it was decreased. Exogenous respiration begin to decrease in its O_2$-uptake at 5, 000r. doses of irradiation, further decrease with increasing of doses unproportionally. 2. It appears that plasma-membrane and nuclear membrane of yeast cells have changed and denatured by gamma-irradiation, as exogenous respiration of glucose had been decreased at a dose of 200, 000r's irradiation. 3. The activity of glucose, alcoholic, lactic, succinic and glutamic deliydrogenase (G.D.H., A.D.H., L.D.H., S.D.11., and GL.D.H.) in the gamma irradaited cells had been assayed by T.T.C.(Triphenyl tetrazolium chloride) method and spectrophotometry, the obtained results were compared with those of normal cells. 4. At a dose of and 10, 000 rentgens' irradiation of gamma ray, the activty of each debydrogenase (G.D.H., A.D.H., L.D.H., ) shows a sharp and highest peak in optical absorbalicy, but each abtivity of S.D.H and Gl.D.H shows its' maximum peak at a dose of 30, 000r. 5. The curve of each dehydrogenase activity was found to be rhythmical according to dose-rate of gamma irradiation. 6. Comparing with activity of debydrogenase each other, the maximum peak in optical absorbency can be arranged according to order as follows; glucose > alcoholoic > lactic > glutamic > succinic, this order is identical to the order of breakdown utility in respiration of normal yeast cells. 7. The activity of dehydrogenase experimented exhibit a resistance against gamma irradiation at lethal dose of cells, and the activity of dehydrogenase are found to be much resistant than those of respiratory system. We may assume that the membrane substrate of mitochondria or cytoplasm had been destructed by gamma-irradiation much more than that of dehydronase system.

• Archives of Craniofacial Surgery
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• v.19 no.3
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• pp.181-189
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• 2018

Background: Autogenous bone grafts have several limitations including donor-site problems and insufficient bone volume. To address these limitations, research on bone regeneration is being conducted actively. In this study, we investigate the effects of a three-dimensionally (3D) printed polycaprolactone (PCL)/tricalcium phosphate (TCP) scaffold on the osteogenic differentiation potential of adipose tissue-derived stem cells (ADSCs) and bone marrow-derived stem cells (BMSCs). Methods: We investigated the extent of osteogenic differentiation on the first and tenth day and fourth week after cell culture. Cytotoxicity of the 3D printed $PCL/{\beta}-TCP$ scaffold was evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay, prior to osteogenic differentiation analysis. ADSCs and BMSCs were divided into three groups: C, only cultured cells; M, cells cultured in the 3D printed $PCL/{\beta}-TCP$ scaffold; D, cells cultured in the 3D printed $PCL/{\beta}-TCP$ scaffold with a bone differentiation medium. Alkaline phosphatase (ALP) activity assay, von Kossa staining, reverse transcription-polymerase chain reaction (RT-PCR), and Western blotting were performed for comparative analysis. Results: ALP assay and von Kossa staining revealed that group M had higher levels of osteogenic differentiation compared to group C. RT-PCR showed that gene expression was higher in group M than in group C, indicating that, compared to group C, osteogenic differentiation was more extensive in group M. Expression levels of proteins involved in ossification were higher in group M, as per the Western blotting results. Conclusion: Osteogenic differentiation was increased in mesenchymal stromal cells (MSCs) cultured in the 3D printed PCL/TCP scaffold compared to the control group. Osteogenic differentiation activity of MSCs cultured in the 3D printed PCL/TCP scaffold was lower than that of cells cultured on the scaffold in bone differentiation medium. Collectively, these results indicate that the 3D printed PCL/TCP scaffold promoted osteogenic differentiation of MSCs and may be widely used for bone tissue engineering.

• Korean Journal of Food Science and Technology
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• v.42 no.6
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• pp.768-772
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• 2010

In this study, the neuronal cell protective effects of methanol extract from cheonggukjang were evaluated. The proximate composition and total phenolics of the methanol extract were 40.95% crude protein, 22.49% crude fat, 15.99% nitrogen free extract, 7.91% moisture, 6.74% crude ash, 5.92% crude fiber, and 28.43 mg/g of total phenolics. Intracellular ROS accumulation resulting from $H_2O_2$ treatment of PC12 cells was significantly reduced when methanol extract was present in the media compared to PC12 cells treated with $H_2O_2$ only. In a cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium-bromide (MTT), the methanol extract showed protective effects against $H_2O_2$-induced neurotoxicity, and lactate dehydrogenase (LDH) release into the medium was also inhibited. Furthermore, the inhibitory effect of the methanol extract against acetylcholinesterase was dose-dependent.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.58 no.1
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• pp.20-27
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• 2013

Proximate composition, total phenolics and total flavonoids level, DPPH radical scavenging activity, and cytotoxicity were determined in the methanol extracts of different plant parts of Youngia sonchifolia at reproductive growth stage. Crude protein and crude fat were present as the highest amount in flowers, and crude fiber in the stems and roots. The highest content of phenolics [mg ferulic acid equivalents (FAE) $kg^{-1}$ dry weight (DW)] was found in flowers (highest) and followed by leaves, stems and roots (lowest). Flavonoids [mg rutin equivalents $kg^{-1}$ DW] level, however, showed the highest in leaf extracts and lowest in root extracts. The antioxidant potential of the methanol extracts from the plants dose-dependently increased DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical scavenging activity (%). DPPH radical scavenging activity were highest in root extracts ($IC_{50}=1,135.6\;mg\;kg^{-1}$) and followed by leaf, stem and flower extracts. By way of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, methanol extracts of roots showed the highest anticancer activity on human cancer cell line Calu-6 for human pulmonary carcinoma ($IC_{50}=196.3\;mg\;kg^{-1}$) and HCT-116 for human colon carcinoma ($IC_{50}=623.6\;mg\;kg^{-1}$).

• Journal of the Korean Applied Science and Technology
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• v.31 no.3
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• pp.440-445
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• 2014

The purpose of this study was to investigate anti-inflammatory and antioxidant effects of essential oil from seed of Zanthoxylum schinifolium on cultured RAW 264.7 cell line. Antioxidant activity of essential oil was evaluated by two different assays as 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and super oxide dismutase (SOD) like activities. This essential oil was tested for cell viability on RAW 264.7 cell line by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. The effects of anti-inflammatory on LPS-induced RAW 264.7 cell line was studied by the content of nitric oxide (NO) and prostagladin $E_2$ ($PGE_2$) in cells. The essential oil of seed from Zanthoxylum schinifolium obtained dose-dependently increased the scavenging activity on DPPH radical scavenging activity and SOD like activity. The essential oil showed low cytotoxicity as more than 98% cell viability in $40{\mu}g/mL^{-1}$ concentration. The essential oil of seed extracted from Zanthoxylum schinifolium presented obvious effect on inflammation. These results suggest that essential oil of seed from Zanthoxylum schinifolium may have value as the potential anti-inflammatory effects by decreasing the action of NO and $PGE_2$ and preventing the activation of oxidative.

• Applied Chemistry for Engineering
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• v.32 no.6
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• pp.647-652
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• 2021

In this study, Saussurea Lappa roots were extracted using ethanol and n-hexane, and also the effects on proliferation of human hair dermal papilla cells and fibroblast and related signaling pathway were evaluated. 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyl tetrazolium bromide (MTT) assay was conducted for cell proliferation effect of Saussurea Lappa root extract, and extracellular signal-related kinase (ERK), serine/threonine protein kinase (Akt), wingless-related integration site (Wnt)/𝛽-catenin signaling pathway, and 5𝛼-reductase expression through western blot analysis were measured. Saussurea Lappa root extract significantly increased human hair dermal papilla cells and propagation of fibroblast, promoted phosphorylation of ERK and Akt that get involved in cell proliferation. Additionally, Saussurea Lappa root extract significantly decreased promotion of Akt phosphorylation and cell proliferation by MEK/ERK inhibitor PD98059 and PI3K/Akt inhibitor LY294002. Also, Saussurea Lappa root extract induced intranuclear 𝛽-catenin accumulation by promoting phosphorylation of 𝛽-catenin (Ser552, 675) through phosphorylation of GSK-3𝛽 (Ser9), and suppressed activation of 5𝛼-reductase type I and II. Overall, Saussurea Lappa root induces cell proliferation through vitalization of ERK and Akt route of human hair dermal papilla cells and fibroblast and apoptosis defense mechanism, and can be helpful in hair loss prevention and hair growth by vitalizing the 𝛽-catenin signaling pathway and inhibiting activation of 5𝛼-reductase, which can be used as a potential hair care products.

• Animal Bioscience
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• v.34 no.5
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• pp.801-810
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• 2021

Objective: microRNAs (miRNAs) can play a role in a variety of physiological and pathological processes, and their role is achieved by regulating the expression of target genes. Our previous high-throughput sequencing found that ssc-miR-185 plays an important regulatory role in piglet diarrhea, but its specific target genes and functions in intestinal porcine epithelial cell (IPEC-J2) are still unclear. We intended to verify the target relationship between porcine miR-185 and cell division cycle 42 (CDC42) gene in IPEC-J2 and to explore the effect of miR-185 on the proliferation of IPEC-J2 cells. Methods: The TargetScan, miRDB, and miRanda software were used to predict the target genes of porcine miR-185, and CDC42 was selected as a candidate target gene. The CDC42-3' UTR-wild type (WT) and CDC42-3'UTR-mutant type (MUT) segments were successfully cloned into pmirGLO luciferase vector, and the luciferase activity was detected after co-transfection with miR-185 mimics and pmirGLO-CDC42-3'UTR. The expression level of CDC42 was analyzed using quantitative polymerase chain reaction and Western blot. The proliferation of IPEC-J2 was detected using cell counting kit-8 (CCK-8), methylthiazolyldiphenyl-tetrazolium bromide (MTT), and 5-ethynyl-2'-deoxyuridine (EdU) assays. Results: Double enzyme digestion and sequencing confirmed that CDC42-3'UTR-WT and CDC42-3'UTR-MUT were successfully cloned into pmirGLO luciferase reporter vector, and the luciferase activity was significantly reduced after co-transfection with miR-185 mimics and CDC42-3'UTR-WT. Further we found that the mRNA and protein expression level of CDC42 were down-regulated after transfection with miR-185 mimics, while the opposite trend was observed after transfection with miR-185 inhibitor (p<0.01). In addition, the CCK-8, MTT, and EdU results demonstrated that miR-185 promotes IPEC-J2 cells proliferation by targeting CDC42. Conclusion: These findings indicate that porcine miR-185 can directly target CDC42 and promote the proliferation of IPEC-J2 cells. However, the detailed regulatory mechanism of miR-185/CDC42 axis in piglets' resistance to diarrhea is yet to be elucidated in further investigation.