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Isolation and Culture of Purified Aortic Endothelial Cells Derived from Alpha 1, 3-galactosyltransferase-deficient Pigs (이종 장기이식 및 조직 공학을 위한 Alpha gal 유전자 결손돼지(1, 3-galactosyltransferase-deficient pigs)에서 혈관내피세포(aortic endothelial cells)의 구축)

  • Ock, Sun A;Lim, Malgum;Kim, Yeongji;Ullah, Imran;Shin, Yurianna;Kim, Youngim;Oh, Keon Bong;Hwang, Seongsoo;Hur, Tai-Young;Lee, Seunghoon;Im, Gi-Sun
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.87-94
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    • 2017
  • Tissue engineering (TE) has been developed to create functional organs and tissue by combining 3D matrix and cells in vitro. Vascularization and angiogenesis are utmost important for supply of nutrients and oxygen in tissue engineered organs. The present study was performed to isolate and characterize primary endothelial cells (EC) from aorta of alpha 1, 3-enzyme galactosyltransferase knock out (GalT KO) pig, to minimize immune rejection and analyze body immune system for future xenotransplantation studies. Isolation of primary EC from aorta were performed by incubation with dispase for 8-10 min at $37^{\circ}C$. Primary EC were cultured in EC growth medium on different extra cellular matrix (ECM), either collagen or gelation. Primary EC exhibits morphological characteristics and showed positive expressions of EC specific marker proteins i.e. PECAM1, KDR and VWF despite of their ECM surface; however, on collagen based surface they showed increase in mRNA level analyzed by qPCR. Primary EC cultured on collagen were sorted by flow cytometer using KDR marker and cultured as KDR positive cells and KDR negative cells, respectively. KDR positive cells showed dramatically increased in PECAM1 and VWF level as compared to KDR negative cells. Based on the above results, primary EC derived from GalT KO are successfully isolated and survived continuously in culture without becoming overgrown by fibroblast. Therefore, they can be utilize for xeno organ transfer, tissue engineering, and immune rejection study in future.

Stage specific transcriptome analysis of liver tissue from a crossbred Korean Native Pig (KNP × Yorkshire)

  • Kumar, Himansu;Srikanth, Krishnamoorthy;Park, Woncheol;Lee, Kyung-Tai;Choi, Bong-Hwan;Kim, Jun-Mo;Lim, Dajeong;Park, Jong-Eun
    • Journal of Biomedical and Translational Research
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    • v.19 no.4
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    • pp.116-124
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    • 2018
  • Korean Native Pig (KNP) has a uniform black coat color, excellent meat quality, white colored fat, solid fat structure and good marbling. However, its growth performance is low, while the western origin Yorkshire pig has high growth performance. To take advantage of the unique performance of the two pig breeds, we raised crossbreeds (KNP ${\times}$ Yorkshire to make use of the heterotic effect. We then analyzed the liver transcriptome as it plays an important role in fat metabolism. We sampled at two stages: 10 weeks and at 26 weeks. The stages were chosen to correspond to the change in feeding system. A total of 16 pigs (8 from each stage) were sampled and RNA sequencing was performed. The reads were mapped to the reference genome and differential expression analysis was performed with edgeR package. A total of 324 genes were found to be significantly differentially expressed (${\left|log2FC\right|}$ > 1 & q < 0.01), out of which 180 genes were up-regulated and 144 genes were down-regulated. Principal Component Analysis (PCA) showed that the samples clustered according to stages. Functional annotation of significant DEGs (differentially expressed genes) showed that GO terms such as DNA replication, cell division, protein phosphorylation, regulation of signal transduction by p53 class mediator, ribosome, focal adhesion, DNA helicase activity, protein kinase activity etc. were enriched. KEGG pathway analysis showed that the DEGs functioned in cell cycle, Ras signaling pathway, p53 signaling pathway, MAPK signaling pathway etc. Twenty-nine transcripts were also part of the DEGs, these were predominantly Cys2His2-like fold group (C2H2) family of zinc fingers. A protein-protein interaction (PPI) network analysis showed that there were three highly interconnected clusters, suggesting an enrichment of genes with similar biological function. This study presents the first report of liver tissue specific gene regulation in a cross-bred Korean pig.

A Case Study of Spatial CAD Education in Blended Learning Environment (혼합형 학습(Blended Learning) 환경에서의 공간디자인 CAD 수업 사례연구)

  • Hwang, Ji Hyoun;Lim, Haewon
    • The Journal of the Korea Contents Association
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    • v.21 no.10
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    • pp.115-126
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    • 2021
  • The purpose of this study is to closely analyze the case of blended-learning in order to provide a diverse and flexible learning environment while maintaining the nature of face-to-face classes, and to identify the learning environment that supports blended-learning in each class step and the educational experience of students. The experience and satisfaction of blended learning were investigated in various ways: course evaluation, LMS activity evaluation, and questionnaire before and after the class. As a result, the blended-learning is better than the traditional face-to-face classes, in providing real-time feedback, opportunities for various interactions, and textual conversations, anytime and anywhere. In addition, as a result of the preliminary survey, as a measure to solve the opinion that concentration was reduced due to problems such as networks and felt uncomfortable in the communication part, the theory and lectures of the design practice class were conducted non-face-to-face. The individual Q&A and feedback were conducted face-to-face and non-face-to-face. As a result of the follow-up survey, it was found that concentration and efficiency could be improved. This opens up possibilities for active use of the online environment in design practice classes.

Relationship between DNA mismatch repair and CRISPR/Cas9-mediated knock-in in the bovine β-casein gene locus

  • Kim, Seung-Yeon;Kim, Ga-Yeon;You, Hyeong-Ju;Kang, Man-Jong
    • Animal Bioscience
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    • v.35 no.1
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    • pp.126-137
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    • 2022
  • Objective: Efficient gene editing technology is critical for successful knock-in in domestic animals. RAD51 recombinase (RAD51) gene plays an important role in strand invasion during homologous recombination (HR) in mammals, and is regulated by checkpoint kinase 1 (CHK1) and CHK2 genes, which are upstream elements of RAD51 recombinase (RAD51). In addition, mismatch repair (MMR) system is inextricably linked to HR-related pathways and regulates HR via heteroduplex rejection. Thus, the aim of this study was to investigate whether clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 (CRISPR/Cas9)-mediated knock-in efficiency of human lactoferrin (hLF) knock-in vector in the bovine β-casein gene locus can be increased by suppressing DNA MMR-related genes (MSH2, MSH3, MSH6, MLH1, and PMS2) and overexpressing DNA double-strand break (DSB) repair-related genes (RAD51, CHK1, CHK2). Methods: Bovine mammary epithelial (MAC-T) cells were transfected with a knock-in vector, RAD51, CHK1, or CHK2 overexpression vector and CRISPR/sgRNA expression vector to target the bovine β-casein gene locus, followed by treatment of the cells with CdCl2 for 24 hours. After 3 days of CdCl2 treatment, the knock-in efficiency was confirmed by polymerase chain reaction (PCR). The mRNA expression levels of DNA MMR-related and DNA DSB repair-related genes were assessed by quantitative real-time PCR (RT-qPCR). Results: Treatment with CdCl2 decreased the mRNA expression of RAD51 and MMRrelated genes but did not increase the knock-in efficiency in MAC-T cells. Also, the overexpression of DNA DSB repair-related genes in MAC-T cells did not significantly affect the mRNA expression of MMR-related genes and failed to increase the knock-in efficiency. Conclusion: Treatment with CdCl2 inhibited the mRNA levels of RAD51 and DNA MMR-related genes in MAC-T cells. However, the function of MMR pathway in relation to HR may differ in various cell types or species.

OGLE-2019-BLG-0362Lb: A SUPER-JOVIAN-MASS PLANET AROUND A LOW-MASS STAR

  • Chung, Sun-Ju;Yee, Jennifer C.;Udalski, Andrej;Gould, Andrew;Albrow, Michael D.;Jung, Youn Kil;Hwang, Kyu-Ha;Han, Cheongho;Ryu, Yoon-Hyun;Shin, In-Gu;Shvartzvald, Yossi;Zang, Weicheng;Cha, Sang-Mok;Kim, Dong-Jin;Kim, Seung-Lee;Lee, Chung-Uk;Lee, Dong-Joo;Lee, Yongseok;Park, Byeong-Gon;Pogge, Richard W.;Poleski, Radek;Mroz, Przemek;Pietrukowicz, Pawel;Skowron, Jan;Szymanski, Michal K.;Soszynski, Igor;Kozlowski, Szymon;Rybicki, Krzysztof A.;Iwanek, Patryk;Wrona, Marcin;Gromadzki, Mariusz;Ulaczyk, Krzysztof
    • Journal of The Korean Astronomical Society
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    • v.55 no.4
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    • pp.123-130
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    • 2022
  • We present the analysis of a planetary microlensing event OGLE-2019-BLG-0362 with a shortduration anomaly (~0.4 days) near the peak of the light curve, which is caused by the resonant caustic. The event has a severe degeneracy with ∆𝜒2 = 0.9 between the close and the wide binary lens models both with planet-host mass ratio q ≃ 0.007. We measure the angular Einstein radius but not the microlens parallax, and thus we perform a Bayesian analysis to estimate the physical parameters of the lens. We find that the OGLE-2019-BLG-0362L system is a super-Jovian-mass planet $M_p=3.26^{+0.83}_{-0.58}M_J $ orbiting an M dwarf $M_h=0.42^{+0.34}_{-0.23}M_{\odot}$ at a distance $D_L=5.83^{+1.04}_{-1.55}kpc$. The projected star-planet separation is ${\alpha}_{\bot}= 2.18^{+0.58}_{-0.72}AU$, which indicates that the planet lies beyond the snow line of the host star.

Anti-atopic dermatitis effects of Parasenecio auriculatus via simultaneous inhibition of multiple inflammatory pathways

  • Kwon, Yujin;Cho, Su-Yeon;Kwon, Jaeyoung;Hwang, Min;Hwang, Hoseong;Kang, Yoon Jin;Lee, Hyeon-Seong;Kim, Jiyoon;Kim, Won Kyu
    • BMB Reports
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    • v.55 no.6
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    • pp.275-280
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    • 2022
  • The treatment of atopic dermatitis (AD) is challenging due to its complex etiology. From epidermal disruption to chronic inflammation, various cells and inflammatory pathways contribute to the progression of AD. As with immunosuppressants, general inhibition of inflammatory pathways can be effective, but this approach is not suitable for long-term treatment due to its side effects. This study aimed to identify a plant extract (PE) with anti-inflammatory effects on multiple cell types involved in AD development and provide relevant mechanistic evidence. Degranulation was measured in RBL-2H3 cells to screen 30 PEs native to South Korea. To investigate the anti-inflammatory effects of Parasenecio auriculatus var. matsumurana Nakai extract (PAE) in AD, production of cytokines and nitric oxide, activation status of FcεRI and TLR4 signaling, cell-cell junction, and cell viability were evaluated using qRT-PCR, western blotting, confocal microscopy, Griess system, and an MTT assay in RBL-2H3, HEK293, RAW264.7, and HaCaT cells. For in vivo experiments, a DNCBinduced AD mouse model was constructed, and hematoxylin and eosin, periodic acid-Schiff, toluidine blue, and F4/80-staining were performed. The chemical constituents of PAE were analyzed by HPLC-MS. By measuring the anti-degranulation effects of 30 PEs in RBL-2H3 cells, we found that Paeonia lactiflora Pall., PA, and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. show an inhibitory activity of more than 50%. Of these, PAE most dramatically and consistently suppressed cytokine expression, including IL-4, IL-9, IL-13, and TNF-α. PAE potently inhibited FcεRI signaling, which mechanistically supports its basophil-stabilizing effects, and PAE downregulated cytokines and NO production in macrophages via perturbation of toll-like receptor signaling. Moreover, PAE suppressed cytokine production in keratinocytes and upregulated the expression of tight junction molecules ZO-1 and occludin. In a DNCB-induced AD mouse model, the topical application of PAE significantly improved atopic index scores, immune cell infiltration, cytokine expression, abnormal activation of signaling molecules in FcεRI and TLR signaling, and damaged skin structure compared with dexamethasone. The anti-inflammatory effect of PAE was mainly due to integerrimine. Our findings suggest that PAE could potently inhibit multi-inflammatory cells involved in AD development, synergistically block the propagation of inflammatory responses, and thus alleviate AD symptoms.

Green perilla leaf extract ameliorates long-term oxidative stress induced by a high-fat diet in aging mice

  • Edward, Olivet Chiamaka;Thomas, Shalom Sara;Cha, Kyung-Ok;Jung, Hyun-Ah;Han, Anna;Cha, Youn-Soo
    • Nutrition Research and Practice
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    • v.16 no.5
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    • pp.549-564
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    • 2022
  • BACKGROUND/OBJECTIVES: Oxidative stress is caused by an imbalance between harmful free radicals and antioxidants. Long-term oxidative stress can lead to an "exhausted" status of antioxidant defense system triggering development of metabolic syndrome and chronic inflammation. Green perilla (Perilla frutescens) is commonly used in Asian cuisines and traditional medicine in southeast Asia. Green perilla possesses numerous beneficial effects including anti-inflammatory and antioxidant functions. To investigate the potentials of green perilla leaf extract (PE) on oxidative stress, we induced oxidative stress by high-fat diet (HFD) in aging mice. MATERIALS/METHODS: C57BL/6J male mice were fed HFD continuously for 53 weeks. Then, mice were divided into three groups for 12 weeks: a normal diet fed reference group (NDcon), high-fat diet fed group (HDcon), and high-fat diet PE treated group (HDPE, 400 mg/kg of body weight). Biochemical analyses of serum and liver tissues were performed to assess metabolic and inflammatory damage and oxidative status. Hepatic gene expression of oxidative stress and inflammation related enzymes were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: PE improved hepatopathology. PE also improved the lipid profiles and antioxidant enzymes, including hepatic glutathione peroxidase (GPx) and superoxide dismutase (SOD) and catalase (CAT) in serum and liver. Hepatic gene expressions of antioxidant and anti-inflammatory related enzymes, such as SOD-1, CAT, interleukin 4 (IL-4) and nuclear factor erythroid 2-related factor (Nrf2) were significantly enhanced by PE. PE also reduced the levels of hydrogen peroxide (H2O2) and malondialdehyde (MDA) in the serum and liver; moreover, PE suppressed hepatic gene expression involved in pro-inflammatory response; Cyclooxygenase-2 (COX-2), nitric oxide synthase (NOS), interleukin 1 beta (IL-1β), and interleukin 6 (IL-6). CONCLUSIONS: This research opens opportunities for further investigations of PE as a functional food and possible anti-aging agent due to its attenuative effects against oxidative stress, resulting from HFD and aging in the future.

Dissolution Monitoring of Geo-Soluble Mixtures (지반 소실 혼합재의 용해과정 모니터링)

  • Truong, Q. Hung;Byun, Yong-Hoon;Eom, Yong-Hun;Sim, Young-Jong;Lee, Jong-Sub
    • Journal of the Korean Geotechnical Society
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    • v.25 no.10
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    • pp.111-122
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    • 2009
  • Dissolution of some of geo-materials may yield the loss of the soil strength and the settlement of earth structures. The goal of this study is to monitor the several physical behaviors of soluble mixtures during dissolution. Sand-salt mixtures are used to monitor the meso to macro response including the settlements and shear waves. The mixtures of photoelastic and ice disks are used to monitor micro to meso behavior of soluble mixture including the void ratio, force chain, coordination number and horizontal force changes. In the sand-salt mixtures, shear waves are measured by using bender elements in conventional oedometer cells. In the photoelastic disk - ice disk mixtures, micro to meso response are measured by digital images and load cells. The shear wave velocity decreases at the initial stage of the dissolution, and then increases and approaches to asymptotic value. The larger dissoluble particle and the more random packing produces the severe horizontal fore change. After dissolution, the void increases and the coordination number decreases. This study demonstrates that the particle level behavior such as the changes of the force chain, void ratio, and coordination number affects the global behavior such as the change of the shear wave velocity and horizontal force of the system.

An Experimental Study on Electromagnetic Properties in Early-Aged Cement Mortar under Different Curing Conditions (양생조건에 따른 초기재령 시멘트 모르타르의 전자기 특성에 대한 실험적 연구)

  • Kwon, Seung-Jun;Song, Ha-Won;Maria, Q. Feng
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.28 no.5A
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    • pp.737-746
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    • 2008
  • Recently, NDTs (Non-Destructive Techniques) using electromagnetic(EM) properties are applied to the performance evaluation for RC (Reinforced Concrete) structures. Since nonmetallic materials which are cement-based system have their unique dielectric constant and conductivity, they can be characterized and changed with different mixture conditions like W/C (water to cement) ratios and unit cement weight. In a room condition, cement mortar is generally dry so that porosity plays a major role in EM properties, which is determined at early-aged stage and also be affected by curing condition. In this paper, EM properties (dielectric constant and conductivity) in cement mortar specimens with 4 different W/C ratios are measured in the wide region of 0.2 GHz~20 GHz. Each specimen has different submerged curing period from 0 to 28 days and then EM measurement is performed after 4 weeks. Furthermore, porosity at the age of 28 days is measured through MIP (Mercury Intrusion Porosimeter) and saturation is also measured through amount of water loss in room condition. In order to evaluate the porosity from the initial curing stage, numerical analysis based on the modeling for the behavior in early-aged concrete is performed and the calculated results of porosity and measured EM properties are analyzed. For the convenient comparison with influencing parameters like W/C ratios and curing period, EM properties from 5 GHz to 15 GHz are averaged as one value. For 4 weeks, the averaged dielectric constant and conductivity in cement mortar are linearly decrease with higher W/C ratios and they increase in proportion to the square root of curing period regardless of W/C ratios.

Study on the Feasibility of Space Weapon Development Utilizing Active Debris Removal Techniques and Understanding of Space Maneuver Warfare (우주 쓰레기 제거기술을 활용한 우주무기 개발 개연성 고찰 및 우주기동전(Space Maneuver Warfare)의 이해)

  • Seonghwan Choi
    • Journal of Space Technology and Applications
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    • v.3 no.2
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    • pp.165-198
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    • 2023
  • According to the studies recently published through advanced maui optical and space surveillance technologies (AMOS) Conference 2021, LEO conjunction assessment revolves around not on operating satellites but space debris such as rocket bodies and non-operational satellites, hence suggesting a solution through space traffic management. Against this backdrop, the issue of active debris removal (ADR) has emerged to the surface as an international challenge throughout the globe. In step with this, the United Nations General Assembly approved a resolution calling on nations to halt tests of direct-ascent anti-satellites, to which U.S. and twelve other nations included Republic of Korea were original signatories. ADR techniques are also actively being researched in the civil sector, and these commercial services, if successfully developed, could possibly be utilized for military use as well. As such, this paper will help readers' understanding for the current status of ADR techniques, space threat assessments, on-orbit rendezvous and proximity operations by looking at previous cases, reflecting on space-faring nations' ADR techniques and its development probability in relation to space weapons. As a conclusion, this study will propose the needs of developing space propulsion system by understanding Space Maneuver Warfare in preparation for the future space battlefield.