It has been known that ras signaling transduction leads to cell proliferation and migration including various adaptor molecules. Dynamin protein has been implicated in the formation of nascent vesicles in both the endocytic and secretory pathways. Dynamin was classified into three isoforms: dynamin I is only expressed in neuronal tissue, dynamin II is expressed ubiquitously in all tissue but that of dynamin III is confined to testis. We have reported in previous study that Grb2, binding to ras, was associated with dynamin II in NIH3T3 cells. Therefore we have tried to identify the relative expression of dynamin II according to overexpressed ras protein in ras oncogene transfected cells (NIH3T3 (ras)). For the detection of differential expression of dynamin II, we have used immunofluorescent staining and western blot methods in NIH3T3 and NIH3T3 (ras) cells. Next we have described the morphological differences between NIH3T3 and NIH3T3 (ras) cells using SEM and TEM. From these experiments dynamin II was highly expressed in NIH3T3 (ras) cells. NIH3T3 cells was transformed to more spindle shape with many cell process by transfection of ras oncogene. Moreover dynamin II was more concentrated in endocytotic membrane of the NIH3T3 (ras) cells compared to that of NIH3T3 cells. The present results suggested that dynamin II may involve the intermediate messenger in Ras signaling transduction pathway.
Seo, Jung-Soo;Lim, Sang-Uk;Kim, Na-Young;Lee, Sang-Hwan;Oh, Hyun-Suk;Lee, Hyung-Ho;Chung, Joon-Ki
Journal of fish pathology
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v.18
no.1
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pp.67-80
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2005
Phosphoinositide-specific phospholipase $C\delta$$PLC\delta$) plays an important role in many cellular responses and is involved in the production of second messenger. The present study was conducted to obtain the biochemical characteristics of the expressed recombinant $PLC\delta$ in E. coli cloned from Misgurnus mizolepis and partially purified $PLC\delta$ enzymes from liver tissues of M. mizolepis (wild ML-$PLC\delta$). The ML $PLC\delta$ gene was cloned and expressed under the previous report (Kim et al., 2004), and purified the recombinant protein by successive chromatography using $Ni^{2+}$-NTA affinity column and gel iltration FPLC column. The wild ML-$PLC\delta$ protein was solublized with 2 M KCI and purified by successive chromatography on open heparin-Sephagel and analytical TSKgel heparin-5PW. Both the recombinant and wild ML-$PLC\delta$ form of protein showed a concentration-dependent PLC activity to phosphatidylinositol 4,5-bis-phosphate (PIP$_2$) or phosphatidylinositol (PI). Its activity was absolutely $Ca^{2+}$- dependant, which was similar to mammalian $PLC\delta$ isozymes. Maximal PI-hydrolytic activations of recombinant and wild ML- TEX>$PLC\delta$ was at pH 7.0 and pH 7.5, respectively. In addition, the enzymatic activities of recombinant and wild ML-$PLC\delta$ were increased in concentration-dependent manner by detergent, such as sodium deoxycholate SDC), phosphatidylethanolamine (PE) and phosphatidylcholine (PC). The activities decreased in contrast by a polyamine, such as spermine. Western blotting showed that several types of $PLC\delta$ isozymes exist in various organs. Taken together our results, it suggested that the biochemical characteristics of ML-$PLC\delta$ are similar with those of mammalian $PLC\delta1$ and ${\delta}3$ isozymes.
Kim, Da Sol;Lee, Mi Sun;Kim, Han Sol;Lee, Hye Yun;Kim, Oh Yun;Kang, Ye Rin;Sohn, Dong Hyun;Kim, Koanhoi;Park, Young Chul
Journal of Life Science
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v.27
no.2
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pp.217-224
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2017
Carbon monoxide (CO), a reaction product of cytoprotective enzyme heme oxygenase-1 (HO-1), is a gaseous messenger with anti-proliferative, anti-apoptotic, and anti-inflammatory actions in many cell types. Here, we investigated the role of CO on the process of monocyte differentiation induced by phorbol 12-myristate 13-acetate (PMA) in human monocytic THP-1 cells. CORM-2 (tricarbonyldichlororuthenium (II) dimer, $Ru2Cl_4(CO)_6$), a CO-releasing compound, decreased a marked cell adherence with a slight reduction of proliferation in monocytic THP-1 cells treated with PMA. And, CORM-2 significantly inhibited expression of differentiation markers such as CD14, CD11b plus CD18 (macrophage-1 antigen, Mac-1 or complement receptor 3, CR3) and phagocytosis of carboxylate-modified red fluorescent latex beads, in PMA-stimulated THP-1 cells. For the further experiments, differentiation of PMA-treated cells was enhanced after the initial 2 days stimulus by removing the PMA-containing media then incubating the cells in fresh media for a another 4 days. And, we observed the secretion of inflammatory cytokines and phagocytosis in differentiated macrophages. Treatment with CORM-2 significantly abolished the secretion of IL-6, $TNF-{\alpha}$ and phagocytosis using fluorescence-conjugated E. coli (K-12 strain) bioparticles in lipopolysaccharide (LPS)-stimulated differentiated macrophages. In conclusion, these results suggest that CO inhibits the differentiation of monocytic THP-1 cells as well as the activation of differentiated macrophages.
Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-hodgkin lymphoma. Advances in the chemotherapeutic treatment of this disease have improved the outcomes of DLBCL; nonetheless, many patients still die of DLBCL, and therefore, a better understanding of this disease and identification of novel therapeutic targets are urgently required. In a recent gene expression profiling study, PDE (phosphodiesterase) 4B was found to be overexpressed in chemotherapy-resistant tumors. The major function of PDE4B is to inactivate the second messenger cyclic 3',5' monophosphate (cAMP) by catalyzing the hydrolysis of cAMP to 5'AMP. It is known that cAMP induces cell cycle arrest and/or apoptosis in B cells, and PDE4B abolishes cAMP's effect on B cells. However, the mechanism by which PDE4B is overexpressed remains unclear. Here, we show that the aberrant expression of miRNA may be associated with the overexpression of this gene. The PDE4B 3' untranslated region (UTR) has three functional binding sites of miR-23b, as confirmed by luciferase reporter assays. Interestingly, miR-23b-binding sites were evolutionarily conserved from humans to lizards, implying the critical role of PDE4B-miR-23b interaction in cellular physiology. The ectopic expression of miR-2 3b repressed PDE4B mRNA levels and enhanced intracellular cAMP concentrations. Additionally, miR-23b expression inhibited cell proliferation and survival of DLBCL cells only in the presence of forskolin, an activator of adenylyl cyclase, suggesting that miR-23b's effect is via the downregulation of PDE4B. These results together suggest that miR-23b could be a therapeutic target for overcoming drug resistance by repressing PDE4B in DLBCL.
This study confirms the responses of consumers when the composition of emoticon bundles can be selected by individuals in MIM service. This aims to verify that customized bundling is a valid marketing strategy in the MIM emoticon market. Currently, the emoticon bundling used in Korean MIM services is in the form of pure bundling. As a result, Consumers must purchase an entire bundle even though he/she doesn't need to use all the emoticons contained in it. Some researches(e.g. Hitt & Chen, 2005; Wu & Anandalingam, 2002) show that when consumers value only part of the products or services included in pure bundling, customized bundling is much more profitable. In their works, customized bundling is appropriate when marginal costs are near zero. Information goods, such as emoticons, meet the condition. On the other hand, customized bundling increase the choosable options, so it can pose a problem of complexity (Blecker et al., 2004). And consumers may experience information overload(Huffman & Kahn, 1998). Thus, judgement on the necessity to introduce customized bundling needs to be made through empirical analyses in the light of characteristics of the product and the reaction of consumers. Results show that when customized bundling was introduced, consumers' purchase intention and willingness to pay significantly increased. Purchase intention for customized bundles has increased by 0.44 based on the five point Likert scale than the purchase intention for existing pure bundles. The increase in purchase intention for customized bundles was statistically independent of the existing purchasing experience. In addition, the willingness to pay was increased by about 2.8% compared to the price of the existing emoticon bundles in the whole group. The group with experience in purchasing pure bundles were willing to pay 5.9% more than pure bundles. The other group without experience in purchasing pure bundles were willing to buy if they were about 5% cheaper than the existing price. Overall, introducing customized bundling into emoticon bundles can lead to positive consumers responses and be a viable marketing strategy.
The rapid digitalization across all aspects of society since 1990 led to the digitalization of cartoons. As the medium of cartoons moved from paper to the web, a powerful visual culture emerged. An encounter between cartoons and multimedia technologies has helped cartoons evolve into a video culture. Today cartoons are no longer literate culture. It is critical to pay attention to cartoons as an "expanded fest" and as visual and video culture with much broader significance. In this paper, the investigator set out to diagnose the current position of cartoons changing in the rapidly changing digital age and talk about future directions that they should pursue. Thus she discussed cases of changes from 1990 when colleges began to provide specialized education for cartoons and animation to the present day when cartoon and Multimedia Comics fests exist in addition to the digitalization of cartoons. The encounter between new technologies and cartoons broke down the conventional forms of cartoons. The massive appearance of artists that made active use of new technologies in their works, in particular, has facilitated changes to the content and forms of cartoons and the expansion of character uses. The development of high technologies extends influence to the roles of appreciators beyond the artists' works. Today readers voice their opinions about works actively, build a fan base, promote the works and artists they favor, and help them rise to stardom. As artist groups of various genres were formed, the possibilities of new stories and texts and the appearance of diverse styles and world views have expanded the essence of cartoon texts and the overall cartoon system of cartoon culture, industry, education, institution, and technology. It is expected that cartoons and Multimedia Comics will continue to make a contribution as a messenger to reflect the next generation of culture, mediate it, and communicate with it. Today there is no longer a distinction between print and video cartoons. Cartoons will expand in every field through a wide range of forms and styles, given the current situations involving installation concept cartoons, blockbuster digital videos, fancy items, and characters at theme parks based on a narrative. It is therefore necessary to diversify cartoon and Multimedia Comics education in diverse ways. Today educators are faced with a task to bring up future generations of talents who are capable of leading the culture of overall senses based on literate and video culture by incorporating humanities, social studies, and new technology education into their creative artistic abilities.
The Korean society was forcefully merged with the invasion of Japan in 20th century, and traditional culture of Korea was damaged severely by colonization from Japan. After liberation, Korean society experienced drastic social change with Korean War, and industrial economy and democratic system developed as modernization and democratization occurred. However, Korean traditional culture dissolved more severely as Korean society developed industrial economy and democracy. As criticism of existing Western center of society and the emphasis of cultural identity of non-western regions and third-world, world society preferred exchange of culture of diverse nations and people with each other in advent of postmodernism thoughts in mid-late 20th century. If the cultural identity of Korea was dissolving meanwhile, it was needed to be recovered again. Despite the research in Korean history, language, art, architecture was performed to recover cultural identity of Korea, it did not go in-depth with Korean mythology, for Korean mythology is considered as superstition or savage. Mythology shows subconscious group psychology of people who live in certain specific region. Studying Korean mythology is one of the ways to rediscover cultural identity of Korea. In order for Korean mythology to be known to many people, its stories should be told by media. There were movies, plays, drama, and novels produced based on existing Korean mythology as introduction, then these mythical stories are appear in subculture contents such as recent comics, animation, webtoon, games, and light novels. Then population of game players and webtoon readers increased as dissemination of PC and smart phones, and increasing market scale of subculture contents increased a population of consumers of comics, animation, and light novel. Consumers of sub-culture contents were interested as many of these contents were created, base on Korean mythology. Therefore, this paper is written as research on Korean mythology and its signification in sub-cultural contents which were produced base on Korean mythology.
Objetives : Identification of target genes for ethanol in neurons is important for understanding its molecular and cellular mechanism of action and the neuropathological changes seen in alcoholics. The purpose of this study is to identify of altered gene expression after acute treatmet of ethanol in rat gliom cells. Methods : We used high density cDNA microarray chip to measure the expression patterns of multiple genes in cultured rat glioma cells. DNA microarrays allow for the simultaneous measurement of the expression of several hundreds of genes. Results : After comparing hybridized signals between control and ethanol treated groups, we found that treatment with ethanol increased the expression of 15 genes and decreased the expression of 12 genes. Upregulated genes included Orthodenticle(Drosophila) homolog 1, procollagen type II, adenosine A2a receptor, GATA bindning protein 2. Downregulated genes included diacylglycerol kinase beta, PRKC, Protein phosphatase 1, clathrin-associated protein 17, nucleoporin p58, proteasome. Conclusion : The gene changes noted were those related to the regulation of transcription, signal transduction, second messenger systems. modulation of ischemic brain injury, and neurodengeneration. Although some of the genes were previously known to be ethanol responsive, we have for the most part identified novel genes involved in the brain response to ethanol.
Lee, Hyejin;Kim, Jinhee;Park, Jun Yeon;Kang, Ki Sung;Park, Joeng Hill;Hwang, Gwi Seo
Journal of Ginseng Research
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v.41
no.3
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pp.257-267
/
2017
Background: Heat-processed ginseng, sun ginseng (SG), has been reported to have improved therapeutic properties compared with raw forms, such as increased antidiabetic, anti-inflammatory, and antihyperglycemic effects. The aim of this study was to investigate the antiobesity effects of SG through the suppression of cell differentiation and proliferation of mouse 3T3-L1 preadipocyte cells and the lipid accumulation in Caenorhabditis elegans. Methods: To investigate the effect of SG on adipocyte differentiation, levels of stained intracellular lipid droplets were quantified by measuring the oil red O signal in the lipid extracts of cells on differentiation Day 7. To study the effect of SG on fat accumulation in C. elegans, L4 stage worms were cultured on an Escherichia coli OP50 diet supplemented with $10{\mu}g/mL$ of SG, followed by Nile red staining. To determine the effect of SG on gene expression of lipid and glucose metabolism-regulation molecules, messenger RNA (mRNA) levels of genes were analyzed by real-time reverse transcription-polymerase chain reaction analysis. In addition, the phosphorylation of Akt was examined by Western blotting. Results: SG suppressed the differentiation of 3T3-L1 cells stimulated by a mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI), and inhibited the proliferation of adipocytes during differentiation. Treatment of C. elegans with SG showed reductions in lipid accumulation by Nile red staining, thus directly demonstrating an antiobesity effect for SG. Furthermore, SG treatment down-regulated mRNA and protein expression levels of peroxisome proliferator-activated receptor subtype ${\gamma}$ ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein-alpha ($C/EBP{\alpha}$) and decreased the mRNA level of sterol regulatory element-binding protein 1c in MDI-treated adipocytes in a dose-dependent manner. In differentiated 3T3-L1 cells, mRNA expression levels of lipid metabolism-regulating factors, such as amplifying mouse fatty acid-binding protein 2, leptin, lipoprotein lipase, fatty acid transporter protein 1, fatty acid synthase, and 3-hydroxy-3-methylglutaryl coenzyme A reductase, were increased, whereas that of the lipolytic enzyme carnitine palmitoyltransferase-1 was decreased. Our data demonstrate that SG inversely regulated the expression of these genes in differentiated adipocytes. SG induced increases in the mRNA expression of glycolytic enzymes such as glucokinase and pyruvate kinase, and a decrease in the mRNA level of the glycogenic enzyme phosphoenol pyruvate carboxylase. In addition, mRNA levels of the glucose transporters GLUT1, GLUT4, and insulin receptor substrate-1 were elevated by MDI stimulation, whereas SG dose-dependently inhibited the expression of these genes in differentiated adipocytes. SG also inhibited the phosphorylation of Akt (Ser473) at an early phase of MDI stimulation. Intracellular nitric oxide (NO) production and endothelial nitric oxide synthase mRNA levels were markedly decreased by MDI stimulation and recovered by SG treatment of adipocytes. Conclusion: Our results suggest that SG effectively inhibits adipocyte proliferation and differentiation through the downregulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$, by suppressing Akt (Ser473) phosphorylation and enhancing NO production. These results provide strong evidence to support the development of SG for antiobesity treatment.
Recent explosive increase of electronic commerce provides many advantageous purchase opportunities to customers. In this situation, customers who do not have enough knowledge about their purchases, may accept product recommendations. Product recommender systems automatically reflect user's preference and provide recommendation list to the users. Thus, product recommender system in online shopping store has been known as one of the most popular tools for one-to-one marketing. However, recommender systems which do not properly reflect user's preference cause user's disappointment and waste of time. In this study, we propose a novel recommender system which uses data mining and multi-model ensemble techniques to enhance the recommendation performance through reflecting the precise user's preference. The research data is collected from the real-world online shopping store, which deals products from famous art galleries and museums in Korea. The data initially contain 5759 transaction data, but finally remain 3167 transaction data after deletion of null data. In this study, we transform the categorical variables into dummy variables and exclude outlier data. The proposed model consists of two steps. The first step predicts customers who have high likelihood to purchase products in the online shopping store. In this step, we first use logistic regression, decision trees, and artificial neural networks to predict customers who have high likelihood to purchase products in each product group. We perform above data mining techniques using SAS E-Miner software. In this study, we partition datasets into two sets as modeling and validation sets for the logistic regression and decision trees. We also partition datasets into three sets as training, test, and validation sets for the artificial neural network model. The validation dataset is equal for the all experiments. Then we composite the results of each predictor using the multi-model ensemble techniques such as bagging and bumping. Bagging is the abbreviation of "Bootstrap Aggregation" and it composite outputs from several machine learning techniques for raising the performance and stability of prediction or classification. This technique is special form of the averaging method. Bumping is the abbreviation of "Bootstrap Umbrella of Model Parameter," and it only considers the model which has the lowest error value. The results show that bumping outperforms bagging and the other predictors except for "Poster" product group. For the "Poster" product group, artificial neural network model performs better than the other models. In the second step, we use the market basket analysis to extract association rules for co-purchased products. We can extract thirty one association rules according to values of Lift, Support, and Confidence measure. We set the minimum transaction frequency to support associations as 5%, maximum number of items in an association as 4, and minimum confidence for rule generation as 10%. This study also excludes the extracted association rules below 1 of lift value. We finally get fifteen association rules by excluding duplicate rules. Among the fifteen association rules, eleven rules contain association between products in "Office Supplies" product group, one rules include the association between "Office Supplies" and "Fashion" product groups, and other three rules contain association between "Office Supplies" and "Home Decoration" product groups. Finally, the proposed product recommender systems provides list of recommendations to the proper customers. We test the usability of the proposed system by using prototype and real-world transaction and profile data. For this end, we construct the prototype system by using the ASP, Java Script and Microsoft Access. In addition, we survey about user satisfaction for the recommended product list from the proposed system and the randomly selected product lists. The participants for the survey are 173 persons who use MSN Messenger, Daum Caf$\acute{e}$, and P2P services. We evaluate the user satisfaction using five-scale Likert measure. This study also performs "Paired Sample T-test" for the results of the survey. The results show that the proposed model outperforms the random selection model with 1% statistical significance level. It means that the users satisfied the recommended product list significantly. The results also show that the proposed system may be useful in real-world online shopping store.
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