Lactacystin, a microbial natural product synthesized by Streptomyces, has been commonly used as a selective proteasome inhibitor in many studies. Proteasome inhibitors is known to be preventing the proliferation of cancer cells in vivo as well as in vitro. Furthermore, proteasome inhibitors, as single or combined with other anticancer agents, are suggested as a new class of potential anticancer agents. This study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying induction of apoptosis in SCC25 human tongue sqaumous cell carcinoma cell line treated with lactacystin. The viability of SCC25 cells, human normal keratinocytes (HaCaT cells) and human gingiva fibroblasts (HGF-1 cells), and the growth inhibition of SCC25 cells were assessed by MTT assay and clonogenic assay respectively. The hoechst staining, hemacolor staining and TUNEL staining were conducted to observe SCC25 cells undergoing apoptosis. SCC25 cells were treated with lactacystin, and Western blotting, immunocytochemistry, confocal microscopy, FAScan flow cytometry, MMP activity, and proteasome activity were performed. Lactacystin treatment of SCC25 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. Interestingly, lactacytin remarkably revealed cytotoxicity in SCC25 cells but not normal cells. And tested SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA contents, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, the up-regulation of Bax, and the activation of caspase-7, caspase-3, PARP, lamin A/C and DFF45 (ICAD). Flow cytometric analysis revealed that lactacystin resulted in G1 arrest in cell cycle progression which was associated with up-regulation in the protein expression of CDK inhibitors, $p21^{WAF1/CIP1}$ and $p27^{KIP1}$. We presented data indicating that lactacystin induces G1 cell cycle arrest and apoptois via proteasome, mitochondria and caspase pathway in SCC25 cells. Therefore our data provide the possibility that lactacystin could be as a novel therapeutic strategy for human tongue squamous cell carcinoma.
Lee Hyung Sik;Choi Young Min;Kwon Hyuk Chan;Song Yeon Suk
Radiation Oncology Journal
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v.22
no.2
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pp.145-154
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2004
Purpose : To examine whether a synthetic bile acid derivatives (HS-1200) sensitizes the radiation-induced apoptosis in human breast cancer cells (MCF-7) and to investigate the underlying mechanism. Materials and Methods : Human breast cancer cells (MCF-7) in exponential growth phase were treated with HS-1200 for 24 hours at 37$^{\circ}C$ with 5$\%$ CO$_{2}$ in air atmosphere. After removal of HS-1200, cells were irradiated with 2$\~$8 Gy X-ray, and then cultured Ii drug-free media for 24-96 hours. The effect of radiation on the clonogenicity of MCF-7 cells was determined with clonogenic cell survival assay with 16$\mu$M of HS-1200. The induction of apoptosis was determined using agarose gel electrophoresis and Hoechst staining. The expression level of apoptosis-related molecules, such as PARP, Bax, Bcl-2, Bak and AIF, were assayed by Western blotting analysis with 40$\mu$M of HS-1200 combined with 8 Gy irradiation. To examine the cellular location of cytochrome c, bax and AIF immunofluorescent stainings were undertaken. Results : Treatment of MCF-7 cells with 40$\mu$M of HS-1200 combined with 8 Gy irradiation showed several changes associated with enhanced apoptosis by agarose gel electrophoresis and Hoechst staining. HS-1200 combined with 8 Gy irradiation treatment also enhanced production of PARP cleavage products and increased Bax/Bcl-2 ratio by Western blotting. Loss of mitochondrial membrane potential ($\Delta$$\psi$$_{m}$) and increased cytochrome c staining indicated that cytochrome c had been released from the mitochondria in HS-1200 treated cells. Conclusion : We demonstrated that combination treatment with a synthetic chenodeoxycholic acid derivative HS-1200 and irradiation enhanced radiation-induced apoptosis of human breast cancer cells (MCF-7). We suggest that the increased Bax/Bcl-2 ratio In HS-1200 co-treatment group underlies the increased radio sensitivity of MCF-7 cells. Further futures studies are remained elusive.
Hizikia fusiforme is a kind of brown edible seaweed that mainly grows in the temperate seaside areas of the northwest pacific, including Korea, Japan and China, and has been widely used as a health food for hundreds of years. Recently, H. fusiforme has been known to exert pharmacological activities including antioxidant, antimutagenic and anticoagulant activities. However, the molecular mechanisms of H. fusiforme in malignant cells have not been clearly elucidated yet. In this study, the effects of ethyl alcohol extract of H. fusiforme (EAHF) on the anti-proliferative effects of MDA-MB-231 and MCF-7 human breast cancer cells were investigated. EAHF treatment resulted in a concentration-dependent growth inhibition by including apoptosis in MDA-MB-231 cells and G1 phase arrest in MCF-7 cells, which could be proved by MTT assay, DAPI staining, agarose gel electrophoresis and flow cytometry analysis. In MDA-MB-231 cells, the increase in apoptosis induced by EAHF treatment correlated with up-regulation of pro-apoptotic Bax expression. EAHF treatment induced the proteolytic activation of caspase-3 and caspase-9, and a concomitant inhibition of poly (ADP-ribose) polymerase, $\beta$-catenin, phospholipase-${\gamma}1$ protein and DNA fragmentation factor 45/inhibitor of caspase-activated DNase. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of H. fusiforme.
High-molecular-weight glutenin subunits (HMW-GSs) are extremely important determinants of the functional properties of wheat dough. Transgenic rice plants containing a wheat TaGlu-Ax1 gene encoding a HMG-GS were produced from the Korean wheat cultivar ‘Jokyeong’ and used to enhance the bread-making quality of rice dough using the Agrobacterium-mediated co-transformation method. Two expression cassettes with separate DNA fragments containing only TaGlu-Ax1 and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately into the Agrobacterium tumefaciens EHA105 strain for co-infection. Rice calli were infected with each EHA105 strain harboring TaGlu-Ax1 or HPTII at a 3:1 ratio of TaGlu-Ax1 and HPTII. Among 210 hygromycin-resistant T0 plants, 20 transgenic lines harboring both the TaGlu-Ax1 and HPTII genes in the rice genome were obtained. The integration of the TaGlu-Ax1 gene into the rice genome was reconfirmed by Southern blot analysis. The transcripts and proteins of the wheat TaGlu-Ax1 were stably expressed in rice T1 seeds. Finally, the marker-free plants harboring only the TaGlu-Ax1 gene were successfully screened in the T1 generation. There were no morphological differences between the wild-type and marker-free transgenic plants. The quality of only one HMW-GS (TaGlu-Ax1) was unsuitable for bread making using transgenic rice dough. Greater numbers and combinations of HMW and LMW-GSs and gliadins of wheat are required to further improve the processing qualities of rice dough. TaGlu-Ax1 marker-free transgenic plants could provide good materials to make transgenic rice with improved bread-making qualities.
The purpose of this study is to measure the effect of musical activities on children's aggressiveness using percussion playing through case studies and to present the therapeutic programs. Musical activities using percussion playing were organized for three aggressive children. Twenty-one small group sessions were conducted over seven weeks with 30 minutes given each session. Fourth-grade children involved in using Aggressiveness Measuring Tool for Teachers-revised by Gwak Geum-Joo(1992) was selected for case studies. Children's impulsiveness was also tested through self-test measuring tool for impulsiveness-revision of 16 questions used by Cho Hae Yeon (2001) and Lee Joo Shik (2003). As quantitative method, comparative analysis was made between the pre and post test results using measuring tools for aggressiveness and impulsiveness of children as well as between aggressive behaviors occurring in the initial stage of the first three sessions and in the latter stage of the last three sessions. Qualitative method was used at the same time to examine the effect of percussion playing on children. After the musical activities, child A showed reduced Aggressive Measuring Tool scores from 19 to 18, with child B from 23 to 19 and child C from 21 to 18. The results show that occurrence of aggressive behaviors were lowered in the post test. Impulsiveness Measuring Tool scores in the post test were decreased as well in all three children. During the music therapy programs, it was also observed that the frequency of the target behaviors in all three children has reduced more in the latter stage than the initial stage of music therapy. The qualitative findings indicate that the children experienced releasing stress through self-expression after percussion playing. These findings indicate therapeutic effectiveness of music therapy using on percussion playing in reducing aggressiveness of children as well as the effectiveness of percussion as a therapeutic intervention for aggressive children. These results can be adapted and reapplied by teachers in primary schools to approach children with behavior problems, and can present a useful therapeutic approach to therapists practicing in clinical environments.
This thesis is on the purpose of seeking for the possibility of a mechanism in interpreting, analyzing and criticizing cartoons which are applied to the concept of "the gaze" based on "the otherness of vision", which states a pluralistic visual world. It proves that cartoons are in line with other art works that are the subject to "lack and desire"; the gaze, greets reality and acts as an important criterion of analyzing and criticizing the trend in contemporary art, in which the cartoon expresses the gaze in harmony within its work of art. In this thesis, the artist, Kang Doh-ha structuralized ambiguous and difficult forms of art as he has cumulated experimental minds by working in an indie cartoon plane for a long period of time. Among his works of art, he identified the "invisible world" through his piece "Kubrik". Therefore, he represented: a metaphor and a metonymy, an ambiguously situational expression, an intentional and emotional error, the structure of individuality and integration, and finally tension beyond its meaning through use of 'the gaze' that is both the cause and the subject of a desire in the visible world which Lacan academized when he interpreted and analyzed "Kubrik". The concept of the gaze can be used in a variety of ways to display one another's presence in relation each character, revealing a spot of lack by staring back at readers or audiences and furthermore, to analyze and criticize the hidden side of the art piece by critics. The most important details are the artist's gaze, which is seen in the eyes of the analysis and also his or her criticism of the cartoon, which functions as a metaphoric screen in which the subject himself or herself betrays the law of desires thus enabling the violent and cruel reality to be masked and indulged in plays. This will serve as an element that will lead into an art as well as control the degradation to just a piece of enjoyment with the cartoon remaining only within the visual world.
Journal of the Korean Society of Food Science and Nutrition
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v.44
no.11
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pp.1612-1620
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2015
Inflammation is a complex process involving a variety of immune cells, which defend the body from harmful stimuli. However, pro-inflammatory cytokines and inflammatory mediators can also exacerbate diseases such as cancer. Onion peel contains several phenolic compounds, including quercetin at an amount 20 times greater in peel than edible flesh. Therefore, in this study, the anti-inflammatory effects of onion peel ethanol extract (OPEE) were investigated lipopolysaccharide-induced inflammatory response. In our results, NO production decreased in a dose-dependent manner. Secretion of IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ was suppressed by 44%, 53%, and 60% respectively, at $100{\mu}g/mL$. Moreover, OPEE also suppressed expression of COX-2, iNOS, $NF-{\kappa}B$, and MAPKs in a dose-dependent manner. Formation of mice ear edema was reduced at the highest dose tested compared to the control, and reduction of ear thickness was observed in the histological analysis as well. In the acute toxicity test, no morality was observed in mice administered 5,000 mg/kg body weight of OPEE over a 2-week observation period. These results suggest that OPEE may have significant effects on inflammatory factors and be a potential anti-inflammatory material.
Kim, Sun-Q;Shin, Mi-Kyoung;Auh, Q-Schick;Lee, Jin-Yong;Hong, Jung-Pyo;Chun, Yang-Hyun
Journal of Oral Medicine and Pain
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v.32
no.2
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pp.137-150
/
2007
Trees emit phytoncide into atmosphere to protect them from predation. Phytoncide from different trees has its own unique fragrance that is referred to as forest bath. Phytoncide, which is essential oil of trees, has microbicidal, insecticidal, acaricidal, and deodorizing effect. The present study was performed to examine the effect of phytoncide on Porphyromonas gingivalis, which is one of the most important causative agents of periodontitis and halitosis. P. gingivalis 2561 was incubated with or without phytoncide extracted from Hinoki (Chamaecyparis obtusa Sieb. et Zucc.; Japanese cypress) and then changes were observed in its cell viability, antibiotic sensitivity, morphology, and biochemical/molecular biological pattern. The results were as follows: 1. The phytoncide appeared to have a strong antibacterial effect on P. gingivalis. MIC of phytoncide for the bacterium was determined to be 0.008%. The antibacterial effect was attributed to bactericidal activity against P. gingivalis. It almost completely suppressed the bacterial cell viability (>99.9%) at the concentration of 0.01%, which is the MBC for the bacterium. 2. The phytoncide failed to enhance the bacterial susceptibility to ampicillin, cefotaxime, penicillin, and tetracycline but did increase the susceptibility to amoxicillin. 3. Numbers of electron dense granules, ghost cell, and vesicles increased with increasing concentration of the phytoncide, 4. RT-PCR analysis revealed that expression of superoxide dismutase was increased in the bacterium incubated with the phytoncide. 5. No distinct difference in protein profile between the bacterium incubated with or without the phytoncide was observed as determined by SDS-PAGE and immunoblot. Overall results suggest that the phytoncide is a strong antibacterial agent that has a bactericidal action against P. gingivalis. The phytoncide does not seem to affect much the profile of the major outer membrane proteins but interferes with antioxidant activity of the bacterium. Along with this, yet unknown mechanism may cause changes in cell morphology and eventually cell death.
Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Park, Sun-Hee;Park, So-Young;Choi, Hyeun-Deok;Choi, Jung-Su;Jang, Mi-Ran;Im, Moo-Hyeog;Ahn, Dong-Hyun
Journal of the Korean Society of Food Science and Nutrition
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v.46
no.5
/
pp.537-544
/
2017
In this study, the anti-inflammatory effect of Polyopes affinis ethanol extract (PAEE) was investigated using LPS-stimulated RAW 264.7 cells and a croton oil-induced ICR mice model. Treatment with PAEE significantly reduced production of nitric oxide (NO) and pro-inflammatory cytokines [interleukin (IL)-6, tumor necrosis factor $(TNF)-{\alpha}$, and $IL-1{\beta}$] in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. PAEE treatment also reduced expression of inducible NO synthase, cyclooxygenase-2, nuclear $factor-{\kappa}B$, and mitogen-activated protein kinases in LPS-stimulated RAW 264.7 cells. In the croton oil-induced ear edema test, application of PAEE (10~250 mg/kg body weight) reduced ear edema in a dose-dependent manner, and PAEE treatment at 50 mg/kg body weight showed similar inhibitory effects compared with prednisolone (10 mg/kg body weight). Histological analysis revealed reduced dermal thickness and lower number of infiltrated mast cells. These results suggest that PAEE might be used as a promising anti-inflammatory agent for inhibition of LPS-induced inflammation and ear edema formation.
This study was designed to identify the relationship between emotional labor and reaction factor(job satisfaction, fatigue), buffer factor(social support) among dental hygienist. The number of respondents was 50 who work in dental clinic and period of investigation was march 2010 through April 2010. A structured questionnaire was employed to evaluate the level of participants' sociodemographics, job-related factors, health-related behaviors, emotional labor and self-perceived fatigue, job satisfaction, social support. While the mean values of emotional labor were higher than thosewho work in hospital and lower than those who work in service employees. These results suggest that emotional labor was related to an increase in fatigue and strong correlation between involuntary emotional expression, self-perceived fatigue, job satisfaction, social support in dental hygienist. Multiple regression analysis has found that social support and no. of personnel were main factors that influence to the level of emotional labor in dental hygienist. The results of this study suggest that emotional labor was a determinant predictor of self perceived fatigue. Thus, a management program for emotional labor is strongly recommended for promotion of quality of life in dental hygienist and to improve dental service and competitiveness.
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