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Specific Knockdown of Nanog Expression by RNA Interference in P19 Embryonal Carcinoma Stem Cells (P19 배아 암종 줄기세포에서 RNA 간섭에 의한 Nanog 유전자 발현의 특이적 억제)

  • Choi, Seung-Cheol;Lim, Do-Sun
    • Development and Reproduction
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    • v.12 no.2
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    • pp.159-168
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    • 2008
  • Nanog is a newly identified member of the homeobox family of DNA binding transcription factors that functions to maintain the undifferentiated state of stem cells. However, molecular mechanisms underlying the function of Nanog remain largely unknown. To elucidate the regulatory roles of Nanog involved in maintenance of P19 embryonal carcinoma (EC) stem cells, we transfected three small interfering RNA (siRNA) duplexes targeted against different regions of the Nanog gene into P19 cells. The Nanog siRNA-100 duplexes effectively decreased the expression of Nanog up to 30.7% compared to other two Nanog siRNAs, the Nanog siRNA-400 (67.9 %) and -793 (53.0%). When examined by RT-PCR and real-time PCR, the expression of markers for pluripotency such as Fgf4, Oct3/4, Rex1, Sox1 and Yes was downregulated at 48 h after transfection with Nanog siRNA-100. Furthermore, expression of the ectodermal markers, Fgf5 and Isl1 was reduced by Nanog knockdown. By contrast, the expression of other markers for pluripotency such as Cripto, Sox2 and Zfp57 was not affected by Nanog knockdown at this time. On the other hand, the expression of Lif/Stat3 pathway molecules and of the endoderm markers including Dab2, Gata4, Gata6 and the germ cell nuclear factor was not changed by Nanog knockdown. The results of this study demonstrated that the knockdown of Nanog expression by RNA interference in P19 cells was sufficient to modulate the expression of pluripotent markers involved in the self-renewal of EC stem cells. These results provide the valuable information on potential downstream targets of Nanog and add to our understanding of the function of Nanog in P19 EC stem cells.

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Massive pulmonary hemorrhage in enterovirus 71-infected hand, foot, and mouth disease

  • Lee, Dong Seong;Lee, Young Il;Ahn, Jeong Bae;Kim, Mi Jin;Kim, Jae Hyun;Kim, Nam Hee;Hwang, Jong Hee;Kim, Dong Wook;Lee, Chong Guk;Song, Tae Won
    • Clinical and Experimental Pediatrics
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    • v.58 no.3
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    • pp.112-115
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    • 2015
  • Hand, foot, and mouth disease (HFMD) is an acute, mostly self-limiting infection. Patients usually recover without any sequelae. However, a few cases are life threatening, especially those caused by enterovirus 71 (EV71). A 12-month-old boy was admitted to a primary hospital with high fever and vesicular lesions of the mouth, hands, and feet. After 3 days, he experienced 3 seizure episodes and was referred to our hospital. On admission, he was conscious and his chest radiograph was normal. However, 6 hours later, he suddenly lost consciousness and had developed a massive pulmonary hemorrhage that continued until his death. He experienced several more intermittent seizures, and diffuse infiltration of both lung fields was observed on chest radiography. Intravenous immunoglobulin, dexamethasone, cefotaxime, leukocyte-depleted red blood cells, fresh frozen plasma, inotropics, vitamin K, and endotracheal epinephrine were administered. The patient died 9 hours after intubation, within 3 days from fever onset. EV71 subgenotype C4a was isolated retrospectively from serum and nasopharyngeal swab by real-time reverse transcription-polymerase chain reaction. Here, we report a fatal case of EV71-associated HFMD with sudden-onset massive pulmonary hemorrhage and suspected encephalitis.

Exploratory Study of the Potential for Adolescent Children from Multi-Cultural Families to Be Trained as Competent Personnel in Charge of ODA (다문화가정 청소년자녀의 공적개발원조 (Official Development Assistance: ODA) 인력 양성을 위한 탐색적 연구)

  • Park, Mee Sok;Kim, Kyoung A
    • Journal of Family Resource Management and Policy Review
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    • v.20 no.3
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    • pp.95-113
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    • 2016
  • This study investigates the potential for multi-cultural youths, given their innate bilingual and dual culture, to be trained as the competent ODA professionals. For the purpose of collecting information, we organized two different kinds of FGI in 2015. The first FGI was conducted with a group of 10 expert participants from August 17 to September 25. The second FGI was conducted four times, from October 5 to October 23, with 13 adolescents from multi-cultural families from Russia, Mongolia, Vietnam, India, Indonesia, Japan, and China. The semi-structured interview items were passed through a formal review process of the Institutional Ethics Committee. Furthermore, with the participants' pre-consent to recording, all recorded data were categorized through the transcription process. The results are as follows. While the potential for nourishing bilingual and dual cultural capabilities are high for the adolescent children of multi-cultural families, the expert group also emphasized the negative aspects of these capabilities being suppressed due to the relatively oppressed family environment. The expert group therefore suggests the following: building parent awareness of ODA human resources, developing an educational career transition roadmap, and providing institutional incentives for ODA. In addition, to some extent, the researchers were able to detect some positive self-esteem among the adolescent children of multi-cultural families as a result of the bilingual and dual cultural competencies required to be ODA personnel. In sum, the research provides insight into the need for national social support for the adolescent children of multi-cultural families, so that their strengths are not extinguished, in order that they might grow up as competent ODA personnel.

Cancer Prevention with Green Tea and Its Principal Constituent, EGCG: from Early Investigations to Current Focus on Human Cancer Stem Cells

  • Fujiki, Hirota;Watanabe, Tatsuro;Sueoka, Eisaburo;Rawangkan, Anchalee;Suganuma, Masami
    • Molecules and Cells
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    • v.41 no.2
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    • pp.73-82
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    • 2018
  • Cancer preventive activities of green tea and its main constituent, (-)-epigallocatechin gallate (EGCG) have been extensively studied by scientists all over the world. Since 1983, we have studied the cancer chemopreventive effects of EGCG as well as green tea extract and underlying molecular mechanisms. The first part of this review summarizes groundbreaking topics with EGCG and green tea extract: 1) Delayed cancer onset as revealed by a 10-year prospective cohort study, 2) Prevention of colorectal adenoma recurrence by a double-blind randomized clinical phase II trial, 3) Inhibition of metastasis of B16 melanoma cells to the lungs of mice, 4) Increase in the average value of Young's moduli, i.e., cell stiffness, for human lung cancer cell lines and inhibition of cell motility and 5) Synergistic enhancement of anticancer activity against human cancer cell lines with the combination of EGCG and anticancer compounds. In the second part, we became interested in cancer stem cells (CSCs). 1) Cancer stem cells in mouse skin carcinogenesis by way of introduction, after which we discuss two subjects from our review on human CSCs reported by other investigators gathered from a search of PubMed, 2) Expression of stemness markers of human CSCs compared with their parental cells, and 3) EGCG decreases or increases the expression of mRNA and protein in human CSCs. On this point, EGCG inhibited self-renewal and expression of pluripotency-maintaining transcription factors in human CSCs. Human CSCs are thus a target for cancer prevention and treatment with EGCG and green tea catechins.

An Exploring Study for the Sources of Sports Coaching Efficacy (스포츠 코칭효능감의 원천(source)에 대한 탐색적 연구)

  • Kim, In-Woo;Kim, Ji-Sun;Kim, Han-beom
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.3
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    • pp.840-850
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    • 2021
  • The purpose of this study was to phenomenologically explore what experiences sports coaches have felt about coaching efficacy through qualitative research methods, and to derive the sources of coaching efficacy. To achieve the research aim, 15 sports coaches with more than 5 years of experience were selected as a nomadic sampling and in-depth interviews were conducted, and the collected data were analyzed through qualitative data analysis procedures such as transcription, coding, categorization and meaning. As a result, the sources of sports coaching efficacy were established as five factors including coaching success experience, social support, coaching-related player experience, coaching expertise, and player feedback. The derived sources can be used as educational basic data for the development of coaching efficacy.

Anti-tumorigenic Effects of Angelica gigase Nakai Extract on MBA-MB-231 through Regulating Lats1/2 Activation (유방암세포에서 LATS1/2 활성에 의한 당귀 추출물의 항암효과)

  • Kim, Cho-Long;Kim, Nambin;Jeong, Han-Sol;Shin, Yu-Su;Mo, Jung-Soon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.34 no.4
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    • pp.177-183
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    • 2020
  • The Hippo-YAP signaling pathway is critical for cell proliferation, survival, and self-renewal in both Drosophila and mammals. Disorder of Hippo-YAP pathway leads to tumor development, progression and poor prognosis in various cancers. YAP/TAZ are the key downstream effectors of the Hippo pathway and they can be inhibited through LATS1/2, core kinases in the Hippo pathway, mediated phosphorylation. In this study, we investigated the effect of Angelica gigas Nakai extract (AGNE) on Hippo-YAP/TAZ pathway. First, ANGE induced YAP/TAZ phosphorylation and dissociation of the YAP/TAZ-TEAD transcription complex. By qRT-PCR, we found that ANGE inhibits the expression of YAP/TAZ-TEAD target gene, CTGF and CYR61. In addition, the transcriptional activity of YAP/TAZ was not suppressed significantly in LATS1/2 double-knockout (DKO) cells by ANGE compared to LATS1/2 wild-type (WT) cells, which means AGNE inhibits YAP/TAZ signaling through direct action on LATS1/2. Further, it was confirmed that AGNE-induced activation of LATS1/2 inhibited the migration potential of the vector-expressing cells by suppressing YAP/TAZ activity. The reduced migration potential was restored in active YAP-TEAD expressing cells. Taken together, the results of this study indicate that ANGE downregulates YAP/TAZ signaling in cells through the activation of LATS1/2.

Prevention of UV-induced Skin Damage by Activation of Tumor Suppressor Genes p53 and $p14^{ARF}$

  • Petersen, R.;John, S.;Lueder, M.;Borchert, S.
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.338-351
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    • 2003
  • UV radiation is the most dangerous stress factor among permanent environmental impacts on human skin. Consequences of UV exposure are aberrant tissue architecture, alterations in skin cells including functional changes. Nowadays new kinds of outdoor leisure-time activities and changing environmental conditions make the question of sun protection more important than ever. It is necessary to recognize that self-confident consumers do not consider to change their way of life, they demand modern solutions on the basis of new scientific developments. In the past one fundamental principle of cosmetics was the use of physical and organic filter systems against damaging UV-rays. Today new research results demonstrate that natural protecting cell mechanisms can be activated. Suitable biological actives strongly support the protection function not from the surface but from the inside of the cell. A soy seed preparation (SSP) was proven to stimulate natural skin protective functions. The major functions are an increased energy level and the prevention of DNA damage. These functions can I be defined as biological UV protection. The tumor suppressor protein p53 plays a key role in the regulation of DNA repair. p53 must be transferred into the phosphorylated form to work as transcription factor for genes which are regulating the cell cycle or organizing DNA repair. A pretreatment with SSP increases the phosphorylation rate of p53 of chronically UV-irradiated human keratinocytes significantly. According to the same test procedure SSP induces a dramatic increase in the expression of the tumor suppressor protein p14$^{ARF}$ that is supporting the p53 activity by blocking the antagonist of p53, the oncoprotein Mdm2. Mdm2, a ubiquitin E3-ligase, downregulates p53 and at the same time it prevents phosphorylation of p53. The positive influence of the tumor suppressor proteins explains the stimulation of DNA repair and prevention of sunburn cell formation by SSP, which was proven in cell culture experiments. In vivo the increased skin tolerance against UV irradiation by SSP could be confirmed too. We have assumed, that an increased repair potential provides full cell functionality.y.

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Detection of etiological agents of proliferative and necrotizing pneumonia in pigs in Jeju (제주지역 돼지에서 증식성 괴사성 폐렴의 원인체 검출)

  • Kim, Jae-Hoon;Jung, Ji-Youl;Yang, Hyoung-Seok;Kim, Jae-Hoon
    • Korean Journal of Veterinary Service
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    • v.45 no.1
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    • pp.63-69
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    • 2022
  • Proliferative and necrotizing pneumonia (PNP) is a form of interstitial pneumonia that occurs in post-weaning pigs. In this study, we investigated the presence of swine influenza virus (SIV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2) and Aujeszky's disease virus (ADV) in PNP lesions in Jeju pigs. Based on the histopathologic criteria for PNP, a total of 50 cases were selected in Jeju pigs between 2008 and 2010. Coupled with histopathological examinations, the presence of ADV and SIV by polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR) and PRRSV and PCV2 by immunohistochemical (IHC) methods were investigated. Based on the PCR and RT-PCR methods, ADV and SIV nucleic acids were not detected in all cases. According to IHC, PRRSV was detected in 38 of the 50 cases examined (76%) and PCV2 in 25 cases (50%). PRRSV or PCV2 were detected in 19 (38%) or 6 (12%) cases, respectively. Both PRRSV and PCV2 were identified in other 19 cases (38%). Antigens of PRRSV and PCV2 were commonly observed in the cytoplasm of macrophages and clusters of necrotic cells in alveolar cavities. The results of the present study demonstrate that PRRSV is predominantly associated with PNP in Jeju pigs. Co-infection with PRRSV and PCV2 may enhance the severity of PNP lesions in affected pigs.

A Case Study on the Experience of Science Teacher Participating in Peer Coaching Meetings (동료 장학 모임에 참여한 과학교사의 경험 사례 연구)

  • Chung, Haengnam;Choi, Byungsoon
    • Journal of The Korean Association For Science Education
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    • v.33 no.1
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    • pp.63-78
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    • 2013
  • Purposes of this study were to explore the process of experience that science teachers go through when participating in peer coaching meetings to improve teaching ability and to find out factors that affect each process of experience. The data were collected through recording of peer coaching meetings, videotapes of science class, and interviews. All the data were analyzed after transcription. The results of the study showed that even though Teacher K broke the ice and formed consensus among the peers by developing Content Representation (CoRe) at the beginning of the meetings, he became self-defensive rather than receptive of peers' opinions on the recorded class at the discussion session. But as the peer coaching went on, he realized that peer coaching was not about evaluation but rather on improving his teaching ability. In turn, he was able to look at his teaching in a more objective point of view and accepted suggestions from peer coaching discussion. The self-reflection of Teacher K acted as the key factor in the efforts to improve his teaching ability. He sought the concrete alternatives through the class analysis with fellow teachers and showed major changes in his teaching practice from the language habits, pronunciation, and speed of his speech to the interaction with students and class design. However, there was little change in knowledge of curriculum and assessment due to his strong orientation to improve students' grades as an academic high school teacher. Likewise, it was found that while peer coaching exert a strong influence on instructional methods and strategies of Teacher K, his strong orientation to improve students' grades hinders a balanced development of subcomponents of PCK.

Regulation of Arabidopsis Circadian Clock by De-Etiolated 1 (DET1) Possibly via Histone 3 Acetylation (H3Ac) (히스톤 3 아세틸화(H3Ac)를 통한 De-Etiolated 1 (DET1)의 애기장대 생체시계 조절)

  • Song, Hae-Ryong
    • Journal of Life Science
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    • v.22 no.8
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    • pp.999-1008
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    • 2012
  • The circadian clock is a self-sustaining 24-hour timekeeper that allows organisms to anticipate daily-changing environmental time cues. Circadian clock genes are regulated by a transcriptional-translational feedback loop. In Arabidopsis, LATE ELONGATED HYPOCOTYL (LHY) and CIRCADIAN CLOCK-ASSOCIATED 1 (CCA1) transcripts are highly expressed in the morning. Translated LHY and CCA1 proteins repress the expression of the TIMING OF CAB EXPRESSION 1 (TOC1) transcripts, which peaks in the evening. The TOC1 protein elevates the expression of the LHY and CCA1 transcripts, forming a negative feedback loop that is believed to constitute the oscillatory mechanism of the clock. In mammals, the transcription factor protein CLOCK, which is a central component of the circadian clock, was reported to have an intrinsic histone acetyltransferase (HAT) activity, suggesting that histone acetylation is important for core clock mechanisms. However, little is known about the components necessary for the histone acetylation of the Arabidopsis clock-related genes. Here, I report that DET1 (De-Etiolated1) functions as a negative regulator of a key component of the Arabidopsis circadian clock gene LHY in constant dark phases (DD) and is required for the down-regulation of LHY expression through the acetylation of histone 3 (H3Ac). However, the HATs directly responsible for the acetylation of H3 within LHY chromatin need to be identified, and a link connecting the HATs and DET1 protein is still absent.