• Title/Summary/Keyword: SOX-2

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Identification of Candidate Porcine miRNA-302/367 Cluster and Its Function in Somatic Cell Reprogramming

  • Son, Dong-Chan;Hwang, Jae Yeon;Lee, Chang-Kyu
    • Reproductive and Developmental Biology
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    • v.38 no.2
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    • pp.79-84
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    • 2014
  • MicroRNAs (miRNAs) are approximately 22 nucleotides of small noncoding RNAs that control gene expression at the posttranscriptional level through translational inhibition and destabilization of their target mRNAs. The miRNAs are phylogenetically conserved and have been shown to be instrumental in a wide variety of key biological processes including cell cycle regulation, apoptosis, metabolism, imprinting, and differentiation. Recently, a paper has shown that expression of the miRNA-302/367 cluster expressed abundantly in mouse and human embryonic stem cells (ESCs) can directly reprogram mouse and human somatic cells to induced pluripotent stem cells (iPSCs) efficiently in the absence of any of the four factors, Oct4, Sox2, c-Myc, and Klf4. To apply this efficient method to porcine, we analyzed porcine genomic sequence containing predicted porcine miRNA-302/367 cluster through ENSEMBL database, generated a non-replicative episomal vector system including miRNA-302/367 cluster originated from porcine embryonic fibroblasts (PEF), and tried to make porcine iPSCs by transfection of the miRNA-302/367 cluster. Colonies expressing EGFP and forming compact shape were found, but they were not established as iPSC lines. Our data in this study show that pig miRNA-302/367 cluster could not satisfy requirement of PEF reprogramming conditions for pluripotency. To make pig iPSC lines by miRNA, further studies on the role of miRNAs in pluripotency and new trials of transfection with conventional reprogramming factors are needed.

In vitro Expansion of Umbilical Cord Blood Derived Mesenchymal Stem Cells (UCB-MSCs) Under Hypoxic Conditions

  • Yang, Jungyun;Kwon, Jihye;Kim, Miyeon;Bae, Yunkyung;Jin, Hyejin;Park, Hohyun;Eom, Young Woo;Rhee, Ki-Jong
    • Biomedical Science Letters
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    • v.21 no.1
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    • pp.40-49
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    • 2015
  • Mesenchymal stem cells (MSCs) have the ability to self-renew and differentiate into multi-lineage cells, thus highlighting the feasibility of using umbilical cord blood-derived MSCs (UCB-MSCs) for cell-therapy and tissueengineering. However, the low numbers of UCB-MSC derived from clinical samples requires that an ex vivo expansion step be implemented. As most stem cells reside in low oxygen tension environments (i.e., hypoxia), we cultured the UCBMSCs under 3% $O_2$ or 21% $O_2$ and the following parameters were examined: proliferation, senescence, differentiation and stem cell specific gene expression. UCB-MSCs cultured under hypoxic conditions expanded to significantly higher levels and showed less senescence compared to UCB-MSCs cultured under normoxic conditions. In regards to differentiation potential, UCB-MSCs cultured under hypoxic and normoxic conditions both underwent similar levels of osteogenesis as determined by ALP and von Kossa assay. Furthermore, UCB-MSCs cultured under hypoxic conditions exhibited higher expression of OCT4, NANOG and SOX2 genes. Moreover, cells expanded under hypoxia maintained a stem cell immnunophenotype as determined by flow cytometry. These results demonstrate that the expansion of human UCB-MSCs under a low oxygen tension microenvironment significantly improved cell proliferation and differentiation. These results demonstrate that hypoxic culture can be rapidly and easily implemented into the clinical-scale expansion process in order to maximize UCB-MSCs yield for application in clinical settings and at the same time reduce culture time while maintaining cell product quality.

Specificity Protein 1 Expression Contributes to Bcl-w-Induced Aggressiveness in Glioblastoma Multiforme

  • Lee, Woo Sang;Kwon, Junhye;Yun, Dong Ho;Lee, Young Nam;Woo, Eun Young;Park, Myung-Jin;Lee, Jae-Seon;Han, Young-Hoon;Bae, In Hwa
    • Molecules and Cells
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    • v.37 no.1
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    • pp.17-23
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    • 2014
  • We already had reported that Bcl-w promotes invasion or migration in gastric cancer cells and glioblastoma multiforme (GBM) by activating matrix metalloproteinase-2 (MMP-2) via specificity protein 1 (Sp1) or ${\beta}$-cateinin, respectively. High expression of Bcl-w also has been reported in GBM which is the most common malignant brain tumor and exhibits aggressive and invasive behavior. These reports propose that Bcl-w-induced signaling is strongly associated with aggressive characteristic of GBM. We demonstrated that Sp1 protein or mRNA expression is induced by Bcl-w using Western blotting or RT-PCR, respectively, and markedly elevated in high-grade glioma specimens compared with low-grade glioma tissues using tissue array. However, relationship between Bcl-w-related signaling and aggressive characteristic of GBM is poorly characterized. This study suggested that Bcl-w-induced Sp1 activation promoted expression of glioma stem-like cell markers, such as Musashi, Nanog, Oct4 and sox-2, as well as neurosphere formation and invasiveness, using western blotting, neurosphere formation assay, or invasion assay, culminating in their aggressive behavior. Therefore, Bcl-w-induced Sp1 activation is proposed as a putative marker for aggressiveness of GBM.

Improved Preimplantation Development of Cloned Porcine Embryos through Supplementation of Histone Deacetylase Inhibitor MS-275

  • Fang, Xun;Qamar, Ahmad Yar;Shin, Sang Tae;Cho, Jongki
    • Journal of Veterinary Clinics
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    • v.36 no.5
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    • pp.253-258
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    • 2019
  • The objective of this study was to analyse the effects of MS-275 (Class I and II histone deacetylase inhibitor) supplementation on the development of porcine in-vitro somatic nuclear transfer embryo production. During in-vitro development, early embryos were exposed to different concentrations of MS-275 (0, $5{\mu}M$, $10{\mu}M$, and $20{\mu}M$). In in-vitro culture supplemented group, the blastocyst development rate was significantly enhanced by $10{\mu}M$ concentration than other groups (24.0% vs. 19.3%, 21.8%, 11.5%; P < 0.05). Additionally, the 6 h supplementation group, significantly improved the blastocysts production than 24 h, 48 h and control groups (26.1% vs. 17.0%, 15.2%, 2.8%; P < 0.05). Following supplementation with optimal concentrations and time ($10{\mu}M$-6 h group), the blastocyst production was significantly higher than control (25.7% vs 15.8%; P < 0.05). The optimal concentrations of MS-275 significantly enhanced the percentages of ICM:TE than control (43.6% vs. 38.4%; P < 0.05) accompanied with significantly higher expression levels of reprogramming related genes (POU5F1, Naong, and SOX2). In conclusion, the optimal concentrations of $10{\mu}M$ MS-275 and 6 h supplementation during in-vitro culture can significantly improve the quality of porcine in-vitro somatic nuclear transfer embryos through histone acetylation and epigenetic modification. Increasing the efficiency of clonal animal production will greatly promote the development of animal disease models and xenotransplantation.

Temporal Variability of CH4 Gas Concentration Collected in Sampling Bag (온실가스 시료 보관시간에 따른 CH4 농도 변화)

  • Hong, Yoonjung;Cho, Changsang;Kang, Seongmin;Yun, Hyun-Ki;Jeon, Eui-Chan
    • Journal of Climate Change Research
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    • v.7 no.4
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    • pp.477-484
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    • 2016
  • In general, $CH_4$ concentrations generated in combustion facilities are known to be ppm units. In most cases, $CH_4$ samples are collected in Tedlar bags and transported to laboratories for analysis. Considering this fact, in the present study, an attempt was made to find out how long samples can be stored in cases where they are kept in bags and transported as a preliminary study for sampling. According to the results of the experiment using simulated gases (1 ppm, 5 ppm, 10 ppm) in Tedlar bag, $CH_4$ was safe up to 240 hr which is the full time. In the case of simulated gases are containing 4 kind gases ($N_2$, $CO_2$, $CH_4$, and $N_2O$). Field samples (samples of obtained by collecting combustion facilities' exhaust gases) are known to contain highly reactive substances (for example NOx, SOx, and VOCs) and may affect each other. In the present study, one site sample was secured from each of a bituminous coal combustion facility and an LNG combustion facility and whether the concentrations of $CH_4$ gas would change over time (24 hr, 96 hr, 144 hr, 192 hr) was checked. Since site samples could not be analyzed on the day of collection, an experiment was started 24 hr after the time point of sampling to analyze the samples. As with the results of analysis of the simulated gas (240 hr), the results of analysis using the site sample indicated that it could be stored for the full study period 192 hr. Therefore, it was judged that if 192 hr would be taken after sampling before the sample would be analyzed, the concentration value should be reliable.

β-carotene regulates cancer stemness in colon cancer in vivo and in vitro

  • Lee, Kyung Eun;Kwon, Minseo;Kim, Yoo Sun;Kim, Yerin;Chung, Min Gi;Heo, Seung Chul;Kim, Yuri
    • Nutrition Research and Practice
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    • v.16 no.2
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    • pp.161-172
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    • 2022
  • BACKGROUND/OBJECTIVES: Colorectal cancer (CRC) is the third most common cancer worldwide and has a high recurrence rate, which is associated with cancer stem cells (CSCs). β-carotene (BC) possesses antioxidant activity and several anticancer mechanisms. However, no investigation has examined its effect on colon cancer stemness. MATERIALS/METHODS: CD133+CD44+ HCT116 and CD133+CD44+ HT-29 cells were isolated and analyzed their self-renewal capacity by clonogenic and sphere formation assays. Expressions of several CSCs markers and Wnt/β-catenin signaling were examined. In addition, CD133+CD44+ HCT116 cells were subcutaneously injected in xenograft mice and analyzed the effect of BC on tumor formation, tumor volume, and CSCs markers in tumors. RESULTS: BC inhibited self-renewal capacity and CSC markers, including CD44, CD133, ALDH1A1, NOTCH1, Sox2, and β-catenin in vitro. The effects of BC on CSC markers were confirmed in primary cells isolated from human CRC tumors. BC supplementation decreased the number and size of tumors and delayed the tumor-onset time in xenograft mice injected with CD133+CD44+ HCT116 cells. The inhibitory effect of BC on CSC markers and the Wnt/β-catenin signaling pathway in tumors was confirmed in vivo as well. CONCLUSIONS: These results suggest that BC may be a potential therapeutic agent for colon cancer by targeting colon CSCs.

Study on Equivalent Consumption Minimization Strategy Application in PTI-PTO Mode of Diesel-Electric Hybrid Propulsion System for Ships

  • Lee, Dae-Hong;Kim, Jong-Su;Yoon, Kyoung-Kuk;Hur, Jae-Jung
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.28 no.3
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    • pp.451-458
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    • 2022
  • In Korea, five major ports have been designated as sulfur oxide emission control areas to reduce air pollutant emissions, in accordance with Article 10 of the "Special Act on Port Air Quality" and Article 32 of the "Ship Pollution Prevention Regulations". As regulations against vessel-originated air pollutants (such as PM, CO2, NOx, and SOx) have been strengthened, the Ministry of Oceans and Fisheries(MOF) enacted rules that newly built public ships should adopt eco-friendly propulsion systems. However, particularly in diesel-electric hybrid propulsion systems,the demand for precise control schemes continues to grow as the fuel saving rate significantly varies depending on the control strategy applied. The conventional Power Take In-Power Take Off(PTI - PTO) mode control adopts a rule-based strategy, but this strategy is applied only in the low-load range and PTI mode; thus, an additional method is required to determine the optimal fuel consumption point. The proposed control method is designed to optimize fuel consumption by applying the equivalent consumption minimization strategy(ECMS) to the PTI - PTO mode by considering the characteristics of the specific fuel oil consumption(SFOC) of the engine in a diesel-electric hybrid propulsion system. To apply this method, a specific fishing vessel model operating on the Korean coast was selected to simulate the load operation environment of the ship. In this study, a 10.2% reduction was achieved in the MATLAB/SimDrive and SimElectric simulation by comparing the fuel consumption and CO2 emissions of the ship to which the conventional rule-based strategy was applied and that to which the ECMS was applied.

Isolation and characterization of feline endometrial mesenchymal stem cells

  • Mi-Kyung Park;Kun-Ho Song
    • Journal of Veterinary Science
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    • v.25 no.2
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    • pp.31.1-31.8
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    • 2024
  • Background: Recently, there has been a growing interest in stem cells for human medicine. Limited feline endometrial mesenchymal stem cell (fEM-MSC) research in veterinary medicine necessitates reporting for future feline disease research and therapy. Objectives: This study aimed to isolate fEM-MSCs from feline endometrial tissues and evaluate their morphology, proliferative ability, differentiation ability, and immunophenotype. Methods: Feline endometrial tissues were obtained from the ovariohysterectomies of healthy cats and isolated using an enzymatic method. The morphology and proliferative ability of the isolated cells were assessed using a doubling time (DT) assay from passages 3 to 6 (P3 - P6). We measured pluripotency gene expressions of cells in P2 using quantitative real-time polymerase chain reaction (qRT-PCR). To investigate MSC characteristics, a trilineage differentiation assay was conducted in P4, and cells in P4 were immunophenotyped using flow cytometry. Results: fEM-MSCs showed a typical spindle-shaped morphology under a microscope, and the DT was maintained from P3 to P6. fEM-MSCs could differentiate into adipocytes, osteoblasts, and chondrocytes, and expressed three pluripotency markers (OCT4, SOX2, and NANOG) by qRT-PCR. Immunophenotypic analysis showed that the fEM-MSCs were CD14 -, CD34 -, CD45 -, CD9+, and CD44+. Conclusions: In this study, the feline endometrium was a novel source of MSCs, and to the best of our knowledge, this is the first report on the isolation method and characteristics of fEM-MSCs.

National Management Measures for Reducing Air Pollutant Emissions from Vessels Focusing on KCG Services (선박 대기오염물질 배출 현황 및 저감을 위한 국가 관리 대책 연구: 해양경찰 업무를 중심으로)

  • Lee, Seung-Hwan;Kang, Byoung-Yong;Jeong, Bong-Hun;Gu, Ja-Yeong
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.26 no.2
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    • pp.163-174
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    • 2020
  • Particulate matter levels are rapidly increasing daily, and this can affect human health. Therefore, air pollutant emissions from sea vessels require management. This study evaluates the status of air pollutants, focusing on air pollutant emissions from the vessels of the Korea Coast Guard (KCG), and proposes national management measures to reduce emissions. According to a report recently released (2018) by the National Institute of Environmental Research (NIER), emissions from vessels constituted 6.4 % of the total domestic emissions, including 13.1 % NOx, 10.9 % SOx, and 9.6 % particulate matter (PM10/PM2.5). Among the rates of pollutant emission from vessels, the emission rates of domestic and overseas cargo vessels were the highest (50.6 %); the ratio of fishing boats was 42.6 %. With respect to jurisdictional sea area, 44.1 % of the emissions are from the south sea, including the Busan and Ulsan ports, and 24.8 % of the emissions are from the west sea, including the Gwangyang and Yeosu ports. The KCG inspects boarding lines to manage emission conditions and regulate air pollutant emissions, but it takes time and effort to operate various discharge devices and measure fuel oil standards. In addition, owing to busy ship schedules, inspection documents are limited in terms of management. Therefore, to reduce the air pollutant emissions of such vessels, regulations will be strengthened to check for air pollutants, and a monitoring system based on actual field data using KCG patrol ships will be established, for each sea area, to manage the emissions of such vessels. Furthermore, there is a need for technological development and institutional support for the introduction of environmentally friendly vessels.

Chemical Characteristics of Precipitation in Quercus Forests in Korea and Japan

  • Kim, Min Sik;Takenaka, Chisato;Park, Ho Taek;Chun, Kun Woo
    • Journal of Korean Society of Forest Science
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    • v.96 no.5
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    • pp.503-509
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    • 2007
  • The major objective of this study was to analyze the difference of the chemical characteristics of acid deposition in Quercus forests in Korea and Japan. The pH values of rainfall at the experimental forest of Kangwon National University (KS site) were higher than those at the Foresta Hills in Japan (JP site), and all chemical contents of throughfall and stemflow were much higher than those of rainfall in Quercus forest stands at the KS and JP site. The pH values, $Ca^{2+}$, $NO_3{^-}$ and $SO{_4}^{2-}$ concentration of throughfall and stemflow at the KS site showed seasonal variation. While at the JP site, the same pattern was shown in the pH values of throughfall and stemflow, however, did not show any difference among seasons. Also, the annual input of all nutrients in these two contrasting forests varied seasonally. These results can be used to predict the amounts of air pollutant that are washed off and leached by the rainfall and Yellow Sand (Asian dust), including NOx and SOx acid pollutants transported easterly from China in the spring. Therefore, it is necessary to quantify the inputs of dry and wet deposition throughout a full year to gain a more complete understanding of the effects of acid deposition on the nutrient cycles in these forest ecosystems.