• Nutrition Research and Practice
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• v.12 no.1
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• pp.41-46
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• 2018

BACKGROUND/OBJECTIVES: Exposure of the normal lung tissue around the cancerous tumor during radiotherapy causes serious side effects such as pneumonitis and pulmonary fibrosis. Radioprotectors used during cancer radiotherapy could protect the patient from side effects induced by radiation injury of the normal tissue. Delphinidin has strong antioxidant properties, and it works as the driving force of a radioprotective effect by scavenging radiation-induced reactive oxygen species (ROS). However, no studies have been conducted on the radioprotective effect of delphinidin against high linear energy transfer radiation. Therefore, this study was undertaken to evaluate the radioprotective effects of delphinidin on human lung cells against a proton beam. MATERIALS/METHODS: Normal human lung cells (HEL 299 cells) were used for in vitro experiments. The 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay assessed the cytotoxicity of delphinidin and cell viability. The expression of radiation induced cellular ROS was measured by the 2'-7'-dicholordihydrofluorescein diacetate assay. Superoxide dismutase activity assay and catalase activity assay were used for evaluating the activity of corresponding enzymes. In addition, radioprotective effects on DNA damage-induced cellular apoptosis were evaluated by Western blot assay. RESULTS: Experimental analysis, including cell survival assay, MTT assay, and Western blot assay, revealed the radioprotective effects of delphinidin. These include restoring the activities of antioxidant enzymes of damaged cells, increase in the levels of pro-survival protein, and decrease of pro-apoptosis proteins. The results from different experiments were compatible with each to provide a substantial conclusion. CONCLUSION: Low concentration ($2.5{\mu}M/mL$) of delphinidin administration prior to radiation exposure was radioprotective against a low dose of proton beam exposure. Hence, delphinidin is a promising shielding agent against radiation, protecting the normal tissues around a cancerous tumor, which are unintentionally exposed to low doses of radiation during proton therapy.

• Microbiology and Biotechnology Letters
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• v.49 no.2
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• pp.148-156
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• 2021

Single-chain antibodies against epidermal growth factor receptor variant III (EGFRvIII) are potentially promising agents for developing antibody-based cancer treatment strategies. We described in our previous study the successful expression of an anti-EGFRvIII scFv antibody in Escherichia coli. However, we could also observe the formation of insoluble aggregates in the periplasmic space, limiting the production yield of the active product. In the present study, we investigated the mechanisms by which growth conditions could affect the expression of the soluble anti-EGFRvIII scFv antibody in small-scale E. coli NiCo21(DE3) cultures, attempting to maximize production. The secreted scFv molecules were purified using Ni-NTA magnetic beads and protein characterization was performed using SDS-PAGE and western blot analyses. We used the ImageJ software for protein quantification and determined the antigen-binding activity of the scFv antibody against the EGFRvIII protein. Our results showed that the highest percentage of soluble scFv expression could be achieved under culture conditions that combined low IPTG concentration (0.1 mM), low growth temperature (18℃), and large culture dish surface area. We found moderate-yield soluble scFv production in the culture medium after lactose-mediated induction, which was also beneficial for downstream protein processing. These findings were confirmed by conducting western blot analysis, indicating that the soluble, approximately 30-kDa scFv molecule was localized in the periplasm and the extracellular space. Moreover, the antigen-binding assay confirmed the scFv affinity against the EGFRvIII antigen. In conclusion, our study reveals that low-speed protein expression is preferable to obtain more soluble anti-EGFRvIII scFv protein in an E. coli expression system.

• Environmental Mutagens and Carcinogens
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• v.11 no.2
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• pp.148-160
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• 1991

Localization of isoenzyme of glutathione transferase Pi class was compared in different human tissues by immunohistochemical analysis. Strong enrich-ment of GST-Pi in the epithelial tissues was observed in the granular layer of skin, nipple and esophagus which are vulnerable to exogenous chemicals and in the duct epithelium such as pancreatic, biliary, salibvary, renal tubules as well as in the steroid biosynthesis organs such as theca and granulosa of ovary, leydig cell of testis and zona reticularis of adrenal glands.

• Journal of the Korean Applied Science and Technology
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• v.34 no.3
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• pp.602-612
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• 2017

This study was performed to verify various effects of 70% ethanol extract and distilled water extract of pine leaf such as antioxidant, anti-inflammatory, skin-lightening effects analysis in cells. The result of antioxidant experiment showed that polyphenol content increased concentration dependently in polyphenol and flavonoid effects. The total content of polyphenols and flavonoids was much higher in ethanol extract than water extract. In all the cells of B16F10, RAW264.7 cytotoxicity at each concentration level didn't appear. The result of measuring NO production inhibition showed that it inhibited NO production led to LPS effectively, so anti-inflammatory activity was confirmed. The result of measuring melanin biosynthesis inhibition showed that there was considerably reduction effect, but it performed western blot, and as a result of MITF, Tyrosinase protein revelation, the inhibition of MITF, Tyrosinase revelation was confirmed concentration dependently, Therefore, in this study pine leaf extract was expected to be used as a cosmetic ingredient.

• International Journal of Oral Biology
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• v.35 no.2
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• pp.69-74
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• 2010

The mu opioid receptor (MOR) has been regarded as the main site of interaction with analgesics in major clinical use, particularly morphine. The repressor element-1 silencing transcription factor (REST) functions as a transcriptional repressor of neuronal genes in non-neuronal cells. However, it is expressed in certain mature neurons, suggesting that it may have complex and novel roles. In addition, the interactions between MOR and REST and their functions remain unclear. In this study, we examined the effects of morphine on the expression of REST mRNA and protein in human neuroblastoma NMB cells to investigate the roles of REST induced by MOR activation in neuronal cells. To determine the effects of morphine on REST expression, we performed RT-PCR, real-time quantitative RT-PCR, western blot analysis and radioligand binding assays in NMB cells. By RTPCR and real-time quantitative RT-PCR, the expression of REST was found to be unchanged by either the MOR agonist morphine or the MOR specific antagonist CTOP. By western blot, morphine was shown to significantly inhibit the expression of REST, but this suppression was completely blocked by treatment with CTOP. In the radioligand binding assay, the overexpression of REST led to an increased opioid ligand binding activity of endogenous MOR in the NMB cells. These results together suggest that morphine inhibits the expression of REST in human neuroblastoma cells through a post-transcriptional regulatory mechanism mediated through MOR.

• Biomedical Science Letters
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• v.24 no.3
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• pp.206-212
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• 2018

Depression is a type of mood disorder characterized by hypochondriasis, decreased appetite, and insomnia. Depression is a disease that affects more than 100 million people worldwide. 2-Nonadecanone (NAC) is a bioactive substance that constitutes Fomes fomentarius, and NAC is expected to have an antidepressant effect. By using the forced swimming test (FST), we investigated the effects of treatment with NAC on immobility subacutely in rats after oral dosing once a day for 2 days. Serum levels of cytokine interleukin-1 beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) were determined by enzyme-linked immunosorbent assay (ELISA). Nuclear factor-2 (Nrf-2) and inducible nitric oxide synthases (iNOS) were analyzed by western blot method. NAC dose-dependently decreased immobility in the FST. NAC dosedependently decreased FST-induced increase of cytokine levels, as manifested by significantly stronger effects on $IL-1{\beta}$ and $TNF-{\alpha}$ levels at higher doses than the lowest dose of NAC. Western blot analysis showed that Nrf-2 was significantly lower in the NAC-treated group than in the disease-induced group. The iNOS results were also significantly lower in the NAC-treated group than in the other groups. Considering FST results, the antidepressant effect of NAC is effective. Considering the results of cytokine and protein expression, this anti-depressant effect may be related to the anti-inflammatory effect. Therefore, it can be said that the anti-inflammatory effect of NAC increases the antidepressant effect in the FST experiment.

• Journal of Marine Bioscience and Biotechnology
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• v.15 no.2
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• pp.33-40
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• 2023

This study aimed to investigate the immunomodulatory function of Pyropia yezoensis hydrothermal (water) extract (PYWE) in comparison to the group treated only with lipopolysaccharides (LPS) in RAW264.7 cells. LPS is known to be an inflammatory mediator that activates macrophages, leading to the secretion of nitric oxide (NO), inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) as defense responses. Through enzyme-linked immunoassay and western blot analyses, it was observed that PYWE increased the expression levels of NO, iNOS, TNF-α, and IL-6 in RAW264.7 cells in a dose-dependent manner, although to a lesser extent compared with the group treated with LPS alone. In addition, the study examined the mitogen-activated protein kinases (MAPKs) pathway, which regulates various cellular activities, including gene expression, mitosis, cell differentiation, transformation, survival, and death. The western blot analysis confirmed that PYWE also regulated the MAPKs pathway. Furthermore, the expression levels of immunomodulatory-related factors increased in the group treated with PYWE compared with the control group. Even though the effects of PYWE were usually less strong than those of LPS, the effects of PYWE increased with increasing doses compared to the control group. This suggests that PYWE could be used to control the immune system.

• The Korea Journal of Herbology
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• v.22 no.2
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• pp.17-24
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• 2007

Objectives : Licorice has been commonly used as a detoxification agent. We previously reported that licorice and its component, liquiritigenin, exhibits cytoprotective activity against Pb-induced toxicity. The present study was conducted to evaluate the effect of liquiritigenin on the lead-induced cytotoxicity in PC-12 cells. Methods : PC-12 cells were pre-treated with liquiritigenin, and further incubated with lead 100 ${\mu}M$ for $12^{\sim}48$ hours. The viability of PC-12 cells was measured by MTT assay, and the levels of proteins were analysed by western blot. Results : Severe cytotoxicity was induced and nitric oxide (NO) production was augmented by the exposure of lead. Liquiritigenin protected cells from lead-induced cytoxicity and reduced NO production in a dose-dependent manner. The inhibition of NO production was due to the suppression of iNOS protein via the inhibition of $NF-{\kappa}B$ nuclear translocation, determined by western blot analysis. Conclusions : These results suggest that liquiritigenin may exert cytoprotective effect against lead toxicity by inhibiting NO production.

• Journal of Marine Bioscience and Biotechnology
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• v.2 no.2
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• pp.86-93
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• 2007

Chronic inflammation has been known to have a close relationship with several diseases including periodontitis, colitis, hepatitis and arthritis. Recently anti-inflammatory agents have been developed from marine natural resources. In this study, Ecklonia cava (EC) was found to have anti-inflammatory effect. Ethanolic extract of EC belonging to brown algae exhibited an excellent inhibitory effect on the production of inflammatory mediators such as tumor necrosis factor-${\alpha}$, interleukin-$1{\beta}$, interleukin-6 and prostaglandin $E_2$ by RA W264.7 cells. Furthermore, in reporter gene assay and western blot analysis, EC extract exerted anti-inflammatory effect via inactivation of NF-${\kappa}B$ transcription factor that regulates the expression of these inflammatory mediators in macrophages. In addition, EC extract inhibited the activity of matrix metalloproteinase that play an important role in chronic inflammation. These results suggest that EC extract may provide a pharmaceutical potential in inhibiting chronic inflammation.

• Journal of Life Science
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• v.21 no.4
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• pp.534-541
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• 2011

Retinoic acid (RA), a metabolite of vitamin A, is known to regulate dedifferentiation of rabbit articular chondrocytes. The regulatory mechanism of dedifferentiation by RA is not yet understood. Thus, the effect of RA on the regulation of nitric oxide (NO)-induced dedifferentiation was investigated in rabbit articular chondrocytes. RA caused loss of the differentiated chondrocyte phenotype as demonstrated by inhibition of type II collagen expression and proteoglycan synthesis. RA also accelerated NO-induced dedifferentiation in rabbit articular chondrocytes as detected by expression of type II collagen and Sox-9 using Western blot analysis and production of sulfated proteoglycan using Alcain blue staining. Further, RA potentiated NO-induced activation of ERK. Inhibition of ERK with PD98059 (PD) recovered the expression of type II collagen and Sox-9 and production of sulfate proteoglycan in NO-induced dedifferentiated chondrocytes by RA treatment. Our findings suggest that RA accelerates NO-induced dedifferentiation of rabbit articular chondrocytes via the ERK pathway.