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Mechanism Underlying a Proteasome Inhibitor, Lactacystin-Induced Apoptosis on SCC25 Human Tongue Squamous Cell Carcinoma Cells (사람혀편평상피세포암종세포에서 proteasome 억제제인 lactacystin에 의해 유도된 세포자멸사의 기전에 대한 연구)

  • Baek, Chul-Jung;Kim, Gyoo-Cheon;Kim, In-Ryoung;Lee, Seung-Eun;Kwak, Hyun-Ho;Park, Bong-Soo;Tae, Il-Ho;Ko, Myung-Yun;Ahn, Yong-Woo
    • Journal of Oral Medicine and Pain
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    • v.34 no.3
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    • pp.261-276
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    • 2009
  • 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.

Topographic Placement(Structure) and Macro Benthos Community in Winter for the Shellfish Farm of Namsung-ri, Goheung (고흥 남성리 패류양식장의 지형 구조와 저서생물 현장 조사)

  • Jo, Yeong-Hyun;Kim, Yun;Ryu, Cheong-Ro;Lee, Kyeong-Sig;Lee, In-Tae;Yoon, Han-Sam;Jun, Sue-Kyung
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.16 no.2
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    • pp.175-183
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    • 2010
  • To understand the variation of macro benthos community according to the installation of structure and topographic placement in the shellfish farm on tidal flat, the practical example of the tidal shellfish growing area at Namsung-ri Goheung was observed. The results of the research for the field observation were summarized as follows. (1) The ground gradient of the shellfish farm was very flat below about $1^{\circ}$. The shellfish farm ground took the shape of $\sqcup$ from the shoreline to the place of 150 m seawards, and the shape of $\sqcap$ from there to the low tide line. During ebb tide, the $\sqcup$ shape ground stored the sea water, and the $\sqcap$ shape ground was supposed to act as the effect factor to leak slowly or to prevent the outflow. (2) The oyster shell bag or the type of riprap wall as the boundary in the shellfish farm was classified into five types. The air exposure time and flooding time were 181 and 434 minutes, respectively. (3) In the numerical experiment, the deep-sea water wave coming in the study area had 0.5 m of maximum wave height to show the very stable conditions and the wave direction pattern of S-direction was dominant at Naro great ridge, and SE, SSW and S-direction were distributed strongly around the shellfish farm. (4) By the grain size analysis, the sediment around tidal flat consisted of gravel 0.00~5.81(average 1.70)%, sand 14.15~18.39(average 13.23)%, silt 27.59~47.15(average 30.84)% and clay 35.79~55.73(average 36.19)%, and the sediment type was divided into (g)M(lightly gravelly mud), sM(sandy mud) and gM(gravelly mud) by Folk's diagram. (5) The macro benthos community survey conducted in this site in January, 2010 showed that 1 species of Mollusca, 8 species of Polychaeta and 2 species of Crustacea appeared, and 11 species occupying over 1% of total abundance were dominant.

Formation and Evolution of the Miocene Ipcheon Subbasin in Yangbuk-myeon, Gyeongju, SE Korea (한반도 남동부 경주시 양북면 마이오세 입천소분지의 형성과 발달사)

  • Seong, Changhun;Cheon, Youngbeom;Son, Moon;Sohn, Young Kwan;Kim, Jin-Seop
    • The Journal of the Petrological Society of Korea
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    • v.22 no.1
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    • pp.19-34
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    • 2013
  • The Ipcheon Subbasin is an isolated Miocene basin in SE Korea, which has the geometry of an asymmetric graben elongated in the NE-SW direction. It is in contact with basement rocks by faults and separated from adjacent Waup and Eoil basins by the basement. The strata of the basin fills have an overall homoclinal structure, dipping toward NW or WNW. The basin fills consist of Early Miocene sediments rich in dacitic volcanic and volcaniclastic deposits and Middle Miocene non-volcanic and nonmarine conglomerates intercalated with sand layers, which are distributed in the northeastern and southwestern parts of the basin, respectively. Kinematic analysis of syndepositional conjugate faults in the basin fills indicates WNW-ESE extension of the basin. These features are very similar to those of the adjacent Waup and Eoil basins, indicating that the basin extension was governed by the NE-trending northwestern border faults and that the basin experienced a propagating rifting from NE to SW. Basaltic materials, which occur abundantly in the Eoil Basin, are totally absent in the Ipcheon Subbasin. The observations of the dacitic tuff and tuffaceous mudstone in the subbasin, on slabs and under microscope, suggest that they have lithologies very similar to those of the Yondongri Tuff in the Waup Basin. The Middle Miocene non-volcanic sediments of the Waup and Eoil basins and the Ipcheon Subbasin are distributed consistently in the southwestern part of each basin. It is thus concluded that the extension of the Ipcheon Subbasin began at about 22 Ma together with the Waup Basin and was lulled during the main extension period of the Eoil Basin between 20-18 Ma. At about 17 Ma, the subbasin was re-extended due to the activation of the Yeonil Tectonic Line associated with the propagating rifting toward SW. This event is interpreted to have provided new sedimentation space for the Middle Miocene sediments in the southwestern parts of the Waup and Eoil basins and the Ipcheon Subbasin as well.

Selection of RAPD marker to discriminate the bolting-resistant varieties and commercial dried medicinal materials of Angelica species (당귀 내추대성 품종 및 건재약재 판별을 위한 RAPD marker 선발)

  • Bang, Kyong-Hwan;Yu, Hong-Seob;Koo, Dal-Hoe;Cho, Joon-Hyeong;Park, Hee-Woon;Seong, Nak-Sul;Park, Sang-Il;Kim, Hong-Sig
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.1
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    • pp.46-50
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    • 2002
  • In DNA level, genetic study of Angelica species was firstly conducted to discriminate the bolting-resistant or low bolting variety, so called as Manchu, from other Korea collected lines and also this technuque was applied to identify the origin of commercial dried materials obtained from current oriental medicinal market. By RAPD analysis with 72 primers including sixty of 10-mers and twelve of 20-mers, respectively, three primers, which were related to the bolting resistant traits of Angelica gigas, were identified. Comparing the RAPD bands, URP04 primer showed the 1.7 kb specific band, which seemed to be related to delaying bolting traits, since it was observed only in Jinbu elite lines but not in others. On the other hand, since 1.2 kb band amplified by OPD11 was observed in other collected lines but not in Manchu var. and Jinbu line, this primer also could be considered as a selection marker for identifying bolting resistant or delaying bolting traits. In the same manner, since OPP09 did not show 1 kb major band but produced 0.8 kb and 1.2 kb bands in Manchu var., these three bands amplified by the primer could be considered one of the important key specifying Manchu var. related with the trait of Angelica gigas. OPC02 primer showed the same band patterns in all Korean collected lines, but not in other foreign introduced lines, such as A. sinensis from China, and A. acutiloba from Japan. Since these four RAPD primers, OPD11, OPP09, URP04, and OPC02 showed the specific polymorphisms in Angelica species, thus, these were useful to discriminate the three Angelica species, A. gigas, A. sinensis, and A. acutiloba.

The Effect of Theophylline on Eotaxin mRNA Expression in Pulmonary Epithelial Cell Line A549 (인체 기도상피세포주에서 Theophylline에 의한 Eotaxin mRNA 발현억제)

  • Han, Min-Soo;Yoo, Jee-Hong;Kang, Hong-Mo
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.6
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    • pp.898-908
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    • 2000
  • Background : Eotaxin a CC chemokine specific for eosinophils, is implicated in the pathogenesis of asthma by recruiting eosinophils into the airways. Theophylline has been used for the treatment of asthma and recently was proposed to have an anti-inflammatory action. The aim of this study is to examine whether theophylline may inhibit the eosinophilic airway inflammation by reducing the expression of eotaxin. Methods : The expression of eotaxin mRNA was assessed by Northern analysis in A549 cells 4 h after stimulation with TNF-$\alpha$ or IL-1$\beta$. And then, theophylline was added to A549 cells stimulated with 0.1 ng/mL IL-1$\beta$. Results : Eotaxin mRNA expression rates induced by 0.1, 1, 10 ng/mL TNF-$\alpha$ as compared with $\beta$-actin, were 7%, 22%. 28%, respectively. Eotaxin mRNA expression rates induced by 0.01, 0.1, 1, 10 ng/mL IL-1$\beta$, as compared with $\beta$-actin, were 10%, 42%, 63%, 72%, respectively. Eotaxin mRNA expression rates after the addition of 0, 0.001, 0.01, 0.1 ${\mu}M$ dexamethasone induced by 10 ng/mL TNF-$\alpha$ as compared with $\beta$-actin, were 27%, 18%, 8%, respectively. Eotaxin mRNA expression rate after the addition of 0, 0.001, 0.01, 0.1 ${\mu}M$ dexamethasone induced by 0.1 ng/mL IL-1$\beta$ as compared with $\beta$-actin, were 43%, 47%, 12%, 8%, respectively. Eotaxin mRNA expression rates after the addition of 0, 0.001, 0.01, 0.1, 1, 10 mM theophylline induced by 0.1 ng/mL IL-1$\beta$, as compared with $\beta$-actin, were 48%, 40%, 33%, 22%, 16%, 14%, respectively. Conclusion : These results suggest that theophylline may reduce eosinophil infiltration of the airway at least in part by reducing the expression of eotaxin under the conditions of these experiments.

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Mutations of p53 Tumor Suppressor Gene in Human Lung Cancer Cell Lines (사람 폐암세포주에서 p53 종양억제유전자의 변이)

  • Hong, Weon-Seon;Hong, Seok-Il;Lee, Dong-Soon;Son, Young-Sook;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.6
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    • pp.653-658
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    • 1993
  • Background: Recent advancement of molecular genetics has revealed that malignant transformation of a cell may be a complex multistep process and this process is grouped, in general, into two distinct categories, activation of protooncogenes and inactivation of tumor suppressor genes. This study was focused on the mutation of p53 tumor suppressor gene, because p53 gene mutation is now generally accepted to be one of the most frequent genetic changes in a variety of human cancers. Although lung cancer is one of the common cancers in Korea, the genetic change in the carcinogenesis process is not yet known clearly. To investigate the role of p53 gene mutation in lung cancer, we examined the mutations of exon 4-8 of the p53 gene in humna lung cancer cell lines, because most of the mutations of p53 gene have been reported to develop in exon 4-8. Method: Genomic DNA was obtained by the digestion of proteinase K and the extraction by phenol-chloroform-ethanol method from two human pulmonary adenocarcinoma cell lines, PC-9 and PC-14, and one human small cell lung cancer cell line, H69. To detect the mutations of exon 4-8 of the p53 gene, polymerase chain reaction single-strand conformation polymorphism(PCR-SSCP) analysis was performed with the DNA extracted from the cells. Results: The mutation of p53 gene was present in all three cell lines tested. In PC-9, PC-14 and H69, the altered mobility was detected in exon 7, 7 and 5, respectively. Conclusion: These results suggest that p53 gene mutation plays an important role in certain steps of the carcinogenesis of human non-small cell and small cell lung cancer.

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An Exploration of the Influencing Factors and Development of Effective Models of Science Teacher Efficiency (과학 교사의 효능감 관련 요인 탐색을 통한 과학 교사 효능감 형성 모형 개발)

  • Choi, Sung-Youn;Kim, Sung-Won
    • Journal of The Korean Association For Science Education
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    • v.30 no.6
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    • pp.693-718
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    • 2010
  • This study investigated secondary school science teachers' experiences to explore the influencing factors in science teachers efficiency (STE). The participants, thirty three secondary school science teachers who have more than four years of teaching experience, were interviewed about describing each teacher's experience throughout one's years of teaching. The grounded theory introduced by Strauss and Corbin (1998) was used to analyze the data in this study. The results of paradigm analysis revealed that STE is influenced by 125 concepts, 38 sub-categories, and 16 categories. In a paradigm model, the central phenomenon was 'constructing STE', and the causal condition was 'want to be a teacher' as career choice motivation. The contextual conditions that have an affect on the central phenomenon were 'self awareness of the teacher' and 'social awareness of the teacher.' The mediate conditions, which facilitated or restrained the action/interaction strategies, were 'societal tendency', 'school climate', and 'personal context.' The action/interaction strategies to control the phenomenon were 'following the line,' 'identifying effective teaching strategies,' 'taking teacher education programs,' and 'contributing to school improvement.' The consequences were 'teacher's self awareness', 'challenge,' and 'stagnating in teaching.' The overall conclusion drawn from this research is that, the definition of STE is beliefs in science teachers' capabilities to set up objects in some school teaching context and, organize and execute the course of action required to attain these. Additionally, STE has three dimensions of teacher's behaviors: science instructional efficiency, efficiency in engaging students, and efficiency in managing school conditions. This study offers insight into the nature of STE and theoretical framework. These findings may give science teachers and teacher educators the practical knowledge necessary to build effective training programs and interventions that would help increase STE and facilitate effective teaching.

Cognitive-enhancing Effects of a Fermented Milk Product, LHFM on Scopolamine-induced Amnesia (발효유 산물인 LHFM의 인지기능 개선 효과)

  • Jeon, Yong-Jin;Kim, Jun-Hyeong;Lee, Myong-Jae;Jeon, Woo-Jin;Lee, Seung-Hun;Yeon, Seung-Woo;Kang, Jae-Hoon
    • Korean Journal of Food Science and Technology
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    • v.44 no.4
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    • pp.428-433
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    • 2012
  • Probiotics and their products, such as yogurt and cheese have been widely consumed in many countries with proven health benefits including anti-microbial activity and anti-diarrheal activity. LHFM (Lactobacillus helveticus - fermented milk) is a processed skim milk powder, fermented by a probiotics, L. helveticus IDCC3801. In the present study, we aimed to investigate the neuroprotective effects and the cognitive improvements of LHFM. LHFM itself did not show any cytotoxicity to the human neuroblastoma cell line, SH-SY5Y; however, it dose-dependently protected against glutamate-induced neuronal cell death. LHFM also attenuated scopolamine-induced memory deficit in Y-maze and Morris-water maze. In the analysis of hippocampus after a behavior test, LHFM significantly increased the acetylcholine level and also inhibited acetylcholine esterase activity. Therefore, the raised acetylcholine release partially contributes to the improvement of learning and memory by a treatment with LHFM. These results suggest that LHFM is an effective material for prevention or improvement of cognitive impairments caused by neuronal cell damage and central cholinergic dysfunction.

Characteristic Assessment of Heavy Metals in Dusts Collected by the Air Filtration System at Subway Stations in Daegu, Korea (대구지역 지하철역사 공기여과필터 포집먼지에 함유된 중금속성분의 특성평가)

  • Do, Hwa-Seok;Song, Hee-Bong;Shin, Dong-Chan;Kwak, Jin-Hee;Lee, Myoung-Sook;Yoon, Ho-Suk;Kang, Hye-Jung;Phee, Young-Gyu
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.1
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    • pp.42-50
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    • 2009
  • Samples of subway dust were collected by the air filter system of 30 subway stations on Daegu subway line 1 in January 2008. Samples were sieved below 100 ${\mu}m$, and 14 elements were analyzed using ICP after acid extraction. Results obtained from the source assessment of trace elements using enrichment factor showed that Ca, Fe, K, Mg, Mn, Na, V were influenced by natural sources such as weathered rock and resuspended soil, while Cd, Cr, Cu, Ni, Pb and Zn were influenced by anthropogenic sources such as fuel combustion and waste incineration. Concentrations were remarkably higher in components from natural sources than in components from anthropogenic sources. Anthropogenic sources were significantly affected by indoor dusts than outdoor dusts. Results of pollution indices of heavy metals indicated that indoor dusts were more contaminated with heavy metal ions than outdoor dusts. The correlation analysis among trace elements indicated that components were much correlated in the order of natural sources-anthropogenic sources, anthropogenic sources-anthropogenic sources, natural sources-natural sources in both indoor and outdoor dusts. Trace element components of outdoor dusts were largely correlated than those of indoor dusts. In addition, indoor dusts were significantly affected by outdoor dusts rather than depth from the surface or the average daily number of subway passengers.

Chemical Compositions of the Four Lines of Korean Native Chickens (4계통 재래종 닭고기의 화학적 특성)

  • Lee, Kyu Chul;Lee, Sung-Ki;Kim, Hye Kyung
    • Korean Journal of Poultry Science
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    • v.43 no.2
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    • pp.119-128
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    • 2016
  • This study was conducted to compare nutritional composition and taste-related compounds of breast and thigh meats from four lines of Korean native chickens (KNC) Yeonsan Ogye, Hyunin Black, Hwangbong, and Hoengseong Yakdak. White Leghorn (WL) was used as a control. Fifteen male chickens (three chickens in each line) were grown under same condition and slaughtered at 53 weeks old. The contents of Fe and K in KNC, especially Hwangbong breast meat and thigh meat of Hyunin Black and Hoengseong Yakdak, were higher than WL. The contents of Na were lower in KNC compared with WL regardless of parts (p<0.05). Vitamin A contents were higher in thigh meat of WL and Hyunin Black, and vitamin $B_1$ contents were lower in Hoengseong Yakdak than other lines. Vitamin $B_3$ were higher in breast meat of four lines of KNC and thigh meat of Hoengseong Yakdak than WL. Total amino acid contents were higher in breast meat of KNC than WL. The level of good-tasting amino acids were significantly higher in breast meat of Hoengseong Yakdak and WL, and thigh meat of Hyunin Black and WL than other lines of chickens (p<0.05). The ratio of good-tasting amino acids to bitter tasting amino acids was higher in breast meat of Hoengseong Yakdak and thigh meat of Hyunin Black than WL. IMP contents were higher in Hyunin Black and WL than other lines of chickens. Based on these results, it can be concluded that four lines of KNC may have superior nutritional quality and taste when compared with WL.