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Analysis of MAPK Signaling Pathway Genes in the Intestinal Mucosal Layer of Necrotic Eenteritis-Afflicted Two Inbred Chicken Lines

  • Truong, Anh Duc;Hong, Yeojin;Lee, Janggeun;Lee, Kyungbaek;Lillehoj, Hyun S.;Hong, Yeong Ho
    • Korean Journal of Poultry Science
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    • v.44 no.3
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    • pp.199-209
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    • 2017
  • Mitogen-activated protein kinase (MAPK) signaling pathways play a key role in innate immunity, inflammation, cell proliferation, cell differentiation, and cell death. The main objective of this study was to investigate the expression level of candidate MAPK pathway genes in the intestinal mucosal layer of two genetically disparate chicken lines (Marek's disease-resistant line 6.3 and Marek's disease-susceptible line 7.2) induced with necrotic enteritis (NE). Using high-throughput RNA sequencing, we investigated 178 MAPK signaling pathway related genes that were significantly and differentially expressed between the intestinal mucosal layers of the NE-afflicted and control chickens. In total, 15 MAPK pathway genes were further measured by quantitative real-time PCR(qRT-PCR) and the results were consistent with the RNA-sequencing data. All 178 identified genes were annotated through Gene Ontology and mapped onto the KEGG chicken MAPK signaling pathway. Several key genes of the MAPK pathway, ERK1/2, JNK1-3, p38 MAPK, MAP2K1-4, $NF-{\kappa}B1/2$, c-Fos, AP-1, Jun-D, and Jun, were differentially expressed in the two chicken lines. Therefore, we believe that RNA sequencing and qRT-PCR analysis provide resourceful information for future studies on MAPK signaling of genetically disparate chicken lines in response to pathogens.

Identification and Characterization of Expansins from Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae)

  • Lee, Dae-Weon;Seo, Jong Bok;Kang, Jae Soon;Koh, Sang-Hyun;Lee, Si-Hyeock;Koh, Young Ho
    • The Plant Pathology Journal
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    • v.28 no.4
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    • pp.409-417
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    • 2012
  • We identified two novel expansin (EXP) genes in the expressed sequence tag database of Bursaphelenchus xylophilus, designated as Bx-EXPB2 and -EXPB3. Novel Bx-EXPBs encoded 150 amino acids and their similarities in coding sequence were 70.7-84.0% to the previously reported EXPB1 of B. xylophilus. Bx-EXPB2 and Bx-EXPB3 were clustered with Bx-EXPB1 and Bm-EXPB1, respectively, forming the independent phylogeny with other nematode EXPs. All identified Bx-EXPBs contained the signal peptide and were only expressed during the propagative stage, suggesting that they are secreted to facilitate nematode migration through hosts by loosening cell walls during infection. Quantitative real-time PCR analysis showed that the relative accumulation of Bx-EXPB3 mRNAs was the highest among the three Bx-EXPs examined and the order of mRNA accumulation was as follows: Bx-EXPB3 > Bx-EXPB2 >> Bx-EXPB1. Homology modeling of Bx-EXPBs showed that the structurally optimum template was EXLX1 protein of Bacillus subtilis, whichshared residues essential for catalytic activity with Bx-EXPB1 and Bx-EXPB2 except for Bx-EXPB3. Taken together, Bx-EXPB1 and Bx-EXPB2 may be involved migration through plant tissues and play a role in pathogenesis.

Expression of mRNAs and Proteins of Cyclin A and LATS Genes in Ovary (Cyclin A와 LATS 유전자들의 난소 내 mRNA 및 단백질 발현에 관한 연구)

  • Park, Chang-Eun;Kim, Dae-Jung;Hong, Sung-No
    • Korean Journal of Clinical Laboratory Science
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    • v.40 no.1
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    • pp.31-40
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    • 2008
  • Despite of the importance of the primordial follicle (PMF) recruitment, factors and mechanisms for process are poorly understood. To evaluate expression and role of the follicular transition from PMF to PMF/primary follicles (PMIF) in the present study, we evaluated expression of lats1, lats2, cyclin A1, and cyclin A2 mRNA and protein, and elucidated and role of lats1-cyclin A in the follicular transition from PMF to PRIF. To analysis of differential expression in PMF and PMIF, each stage follicles were collected by day1 and day5 of immuno-compromised rats (ICR) and analyzed by real-time PCR for the genes. For localization of mRNAs and proteins of the genes, in situ hybridization and immunohistochemistry were performed. We confirmed that the lats1, lats2, cyclin A1, and cyclin A2 mRNA were more expressed in PMF than PMIF. Localization of the four genes expression were observed in nuclei of oocytes from the arrested primordial, and in the surrounding granulosa cells of the growing follicles. The mRNA expressions were gradually decreased with follicular development. From immunohistochemistry studies, Cyclin A1 protein expression were observed in oocyte cytoplasmas of early stage follicles, while observed in granulose cells and oocyte nucleoli during growing follicles. This study suggested that the presence of lats gene family might perform negatively regulation of cell proliferation by modulation of the CDC2/Cyclin A complex activity. lats-cyclin A genes in oocytes of the early stage follicles might play a role in the meiotic cell cycle arrest of the primary oocytes at the primordial follicle stage as well as the follicular growth.

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Expressional Analysis of Glucose Transporter Isoforms in the Efferent Ductules of Male Sprague Dawley Rat during Postnatal Development

  • Seo, Hee-Jung;Son, Chan-Wok;Lee, Ki-Ho
    • Reproductive and Developmental Biology
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    • v.33 no.4
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    • pp.211-216
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    • 2009
  • A cell frequently utilizes glucose as a fuel of energy and a major substrate of lipid and protein syntheses. A regulation of glucose movement into and out of the cells is precisely controlled by cooperative works of passive and sodium-dependent active processes. At least 13 glucose cotransporter (Slc2a, GLUT) isoforms involve in passive movement of glucose in cells. The efferent ductules (EDs) play in a number of important functions for maintenance of male fertility. In the present study, using real-time PCR analysis, we determined gene expression of five Slc2a isoforms in the EDs. In addition, we compared expression levels of these Slc2a isoforms according to postnatal development ages, 1 week, 2 weeks, 1 month, and 3 months. Results from the current study showed that expression of Slc2a1, Slc2a3, and Slc2a5 mRNAs reached the highest levels at 1 month of age, followed by a transient decrease at 3 months of age. In addition, the level of Slc2a4 mRNA reminded at steady until 1 month of age and was significantly reduced at 3 months of age, whereas the highest level of Slc2a 8 mRNA was detected at 2 weeks of age. Data from the present study indicate a differential expression of various Slc2a isoforms in the ED according to postnatal ages. Thus, it is believed that glucose movement through the epithelial cells in the ED would be regulated by the coordinated manner among Slc2a isoforms expressed at a given age.

Learning Experience Study of Problem Based Learning on War history (문제중심학습(PBL) 경험연구 -군사학과 전쟁사 강좌 사례를 중심으로-)

  • Kim, Sung Woo
    • Convergence Security Journal
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    • v.13 no.2
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    • pp.101-109
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    • 2013
  • Problem Based Learning(PBL) is one of effective teaching-learning strategy for enhancing military students' motivation, comparing with the other study method. After monitoring military student classes, we conclude that PBL can enhance the effectiveness of learning in War history case study education and finds merits; PBL assists students to play a more active role in the class, induces students to solve problems independently, and makes the learning military situation real case study. The case study is common in social sciences and life sciences. Case studies may be descriptive or explanatory. It is good for War history education. The demerit of PBL is a costly method as students should spend more time and institutions should provide more manpower and materials. This study suggests that more empirical researches on alternative teaching methods, including PBL, to a lecture in War history education.

Engagement classification algorithm based on ECG(electrocardiogram) response in competition and cooperation games (심전도 반응 기반 경쟁, 협동 게임 참여자의 몰입 판단 알고리즘 개발)

  • Lee, Jung-Nyun;Whang, Min-Cheol;Park, Sang-In;Hwang, Sung-Teac
    • Journal of Korea Game Society
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    • v.17 no.2
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    • pp.17-26
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    • 2017
  • Excessive use of the internet and smart phones have become a social issue. The level of engagement has both positive and negative effects such as good performance or indulgence phenomenon, respectively. This study was to develop an algorithm to determine the engagement state based on cardiovascular response. The participants were asked to play a pattern matching game and the experimental design was divided into cooperation and competition task to provide the level of engagement. The correlation between heart rate and amplitude was analyzed according to each task. The regression equation and accuracy were verified by polynomial regression analysis. The results showed that heart rate and amplitude were positively correlated when the task was a game, and negatively correlated when there was a reference task. The accuracy of classifying between game and reference task was 89%. The accuracy between tasks was confirmed to be 76.5%. This study is expected to be used to quantitatively evaluate the level of engagement in real time.

A Disembarking Notification System in Public City Buses using Smart Device and High Frequency

  • Chung, Myoungbeom
    • Journal of the Korea Society of Computer and Information
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    • v.25 no.8
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    • pp.55-63
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    • 2020
  • Recently, people have come to enjoy using movies and music applications, such as YouTube, Netflix, and Watcha Play, on their smart devices while riding public transportation. However, they sometimes miss their destination stations because of these activities. Therefore, in this paper, we propose a disembarking notification system for public city buses using smart devices and high frequencies generated via the buses' speakers. The high frequencies are generated by the public buses' station information broadcasters. Smart devices then analyze the high-frequency signals with their inner microphones, and the proposed application displays a disembarking notification to the user when the user's destination or stop-over station's signal is the same. To evaluate this system's performance, we tested 1,000 real-time disembarking notifications, and the test results showed 98.9% accuracy. Moreover, we compared these results to those using only OpenAPI, and our proposed system featured far better outcomes. Thus, this proposed notification system can prove a useful technology for many people who often use public city buses, as it can notify specific users of their destination stations. Furthermore, this system will become innovative technology for global public transportation by informing users of their desired stations using speakers.

Variable localization of Toll-like receptors in human fallopian tube epithelial cells

  • Amjadi, Fatemehsadat;Zandieh, Zahra;Salehi, Ensieh;Jafari, Reza;Ghasemi, Nasrin;Aflatoonian, Abbas;Fazeli, Alireza;Aflatoonian, Reza
    • Clinical and Experimental Reproductive Medicine
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    • v.45 no.1
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    • pp.1-9
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    • 2018
  • Objective: To determine the localization, expression, and function of Toll-like receptors (TLRs) in fallopian tube epithelial cells. Methods: The localization of TLRs in fallopian tube epithelial cells was investigated by immunostaining. Surprisingly, the intensity of staining was not equal in the secretory and ciliated cells. After primary cell culture of fallopian tube epithelial cells, ring cloning was used to isolate colonies of ciliated epithelial cells, distinct from non-ciliated epithelial cells. The expression of TLRs 1-10 was examined by quantitative real-time polymerase chain reaction, and protein localization was confirmed by immunostaining. The function of the TLRs was determined by interleukin (IL)-6 and IL-8 production in response to TLR2, TLR3, TLR5, TLR7, and TLR9 ligands. Results: Fallopian tube epithelial cells expressed TLRs 1-10 in a cell-type-specific manner. Exposing fallopian tube epithelial cells to TLR2, TLR3, TLR5, TLR7, and TLR9 agonists induced the secretion of proinflammatory cytokines such as IL-6 and IL-8. Conclusion: Our findings suggest that TLR expression in the fallopian tubes is cell-type-specific. According to our results, ciliated cells may play more effective role than non-ciliated cells in the innate immune defense of the fallopian tubes, and in interactions with gametes and embryos.

Improved Resistance to Oxidative Stress by a Loss-of-Function Mutation in the Arabidopsis UGT71C1 Gene

  • Lim, Chae Eun;Choi, Jung Nam;Kim, In A;Lee, Shin Ae;Hwang, Yong-Sic;Lee, Choong Hwan;Lim, Jun
    • Molecules and Cells
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    • v.25 no.3
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    • pp.368-375
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    • 2008
  • Approximately 120 UDP-glycosyltransferases (UGTs), which are classified into 14 distinct groups (A to N), have been annotated in the Arabidopsis genome. UGTs catalyze the transfer of sugars to various acceptor molecules including flavonoids. Previously, UGT71C1 was shown to glycosylate the 3-OH of hydroxycinnamates and flavonoids in vitro. Such secondary metabolites are known to play important roles in plant growth and development. To help define the role of UGT71C1 in planta, we investigated its expression patterns, and isolated and characterized a loss-of-function mutation in the UGT71C1 gene (named ugt71c1-1). Our analyses by quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR), microarray data mining, and histochemical detection of GUS activity driven by the UGT71C1 promoter region, revealed the tissue-specific expression patterns of UGT71C1 with highest expression in roots. Interestingly, upon treatment with methyl viologen (MV, paraquat), ugt71c1-1 plants displayed enhanced resistance to oxidative stress, and ROS scavenging activity was higher than normal. Metabolite profiling revealed that the levels of two major glycosides of quercetin and kaempferol were reduced in ugt71c1-1 plants. In addition, when exposed to MV-induced oxidative stress, eight representative ROS response genes were expressed at lower levels in ugt71c1-1 plants, indicating that ugt71c1-1 probably has higher non-enzymatic antioxidant activity. Taken together, our results indicate that ugt71c1-1 has increased resistance to oxidative stress, suggesting that UGT71C1 plays a role in some glycosylation pathways affecting secondary metabolites such as flavonoids in response to oxidative stress.

Amelioration of Bleomycin-induced Pulmonary Fibrosis of Rats by an Aldose Reductase Inhibitor, Epalrestat

  • Li, Xianwei;Shen, Yuanyuan;Lu, Yining;Yang, Jieren
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.5
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    • pp.401-411
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    • 2015
  • Aldose reductase (AR) is known to play a crucial role in the mediation of diabetic and cardiovascular complications. Recently, several studies have demonstrated that allergen-induced airway remodeling and ovalbumin-induced asthma is mediated by AR. Epalrestat is an aldose reductase inhibitor that is currently available for the treatment of diabetic neuropathy. Whether AR is involved in pathogenesis of pulmonary fibrosis and whether epalrestat attenuates pulmonary fibrosis remains unknown. Pulmonary fibrosis was induced by intratracheal instillation of bleomycin (5 mg/kg) in rats. Primary pulmonary fibroblasts were cultured to investigate the proliferation by BrdU incorporation method and flow cytometry. The expression of AR, TGF-${\beta}_1$, ${\alpha}$-SMA and collagen I was analyzed by immunohistochemisty, real-time PCR or western blot. In vivo, epalrestat treatment significantly ameliorated the bleomycin-mediated histological fibrosis alterations and blocked collagen deposition concomitantly with reversing bleomycin-induced expression up-regulation of TGF-${\beta}_1$, AR, ${\alpha}$-SMA and collagen I (both mRNA and protein). In vitro, epalrestat remarkably attenuated proliferation of pulmonary fibroblasts and expression of ${\alpha}$-SMA and collagen I induced by TGF-${\beta}_1$, and this inhibitory effect of epalrestat was accompanied by inhibiting AR expression. Knockdown of AR gene expression reversed TGF-${\beta}_1$-induced proliferation of fibroblasts, up-regulation of ${\alpha}$-SMA and collagen I expression. These findings suggest that AR plays an important role in bleomycin-induced pulmonary fibrosis, and epalrestat inhibited the progression of bleomycin-induced pulmonary fibrosis is mediated via inhibiting of AR expression.