Production and Breeding of Transgenic Cloned Pigs Expressing Human CD73

Lee, Seung-Chan,Lee, Haesun,Oh, Keon Bong,Hwang, In-Sul,Yang, Hyeon,Park, Mi-Ryung,Ock, Sun-A,Woo, Jae-Seok,Im, Gi-Sun,Hwang, Seongsoo The Korean Society of Developmental Biology 2017 발생과 생식 Vol.21 No.2

One of the reasons to causing blood coagulation in the tissue of xenografted organs was known to incompatibility of the blood coagulation and anti-coagulation regulatory system between TG pigs and primates. Thus, overexpression of human CD73 (hCD73) in the pig endothelial cells is considered as a method to reduce coagulopathy after pig-to-non-human-primate xenotransplantation. This study was performed to produce and breed transgenic pigs expressing hCD73 for the studies immune rejection responses and could provide a successful application of xenotransplantation. The transgenic cells were constructed an hCD73 expression vector under control porcine Icam2 promoter (pIcam2-hCD73) and established donor cell lines expressing hCD73. The numbers of transferred reconstructed embryos were $127{\pm}18.9$. The pregnancy and delivery rate of surrogates were 8/18 (44%) and 3/18 (16%). The total number of delivered cloned pigs were 10 (2 alive, 7 mummy, and 1 died after birth). Among them, three live hCD73-pigs were successfully delivered by Caesarean section, but one was dead after birth. The two hCD73 TG cloned pigs had normal reproductive ability. They mated with wild type (WT) MGH (Massachusetts General Hospital) female sows and produced totally 16 piglets. Among them, 5 piglets were identified as hCD73 TG pigs. In conclusion, we successfully generated the hCD73 transgenic cloned pigs and produced their litters by natural mating. It can be possible to use a mate for the production of multiple transgenic pigs such as ${\alpha}-1,3-galactosyltransferase$ knock-out /hCD46 for xenotransplantation.

A Trend Analysis on Nursing Leadership in Korea

Haesun Lee,Sangwoo Park,Na Bee Yoon,Kyung Hee Lee 한국간호과학회 2021 한국간호과학회 학술대회 Vol.2021 No.10

Aim(s): Nursing leadership has been highlighted as essential competency throughout healthcare system for improving healthcare quality and patient outcomes. This study aimed to analyze trends of nursing leadership in published articles in Korean journals including leadership style and leadership measurement. Method(s): A literature search was conducted using databases including KISS, RISS, DBpia, KMbase, NAL, and NDSL from January 2011 to August 2020. Search terms were "nurse" or "nursing" and "leadership". A total of 64 studies were reviewed. Leadership style of the studies was categorized according to McEwen and Wills"s classification. Result(s): Most of studies were conducted in hospital setting (96.9%) and quantitative studies accounted for 96.9%. Most frequent leadership style was authentic leadership (26.7%); unit manager accounted for 70.1% of the subject of leadership. The commonly used leadership measures were Authentic Leadereship Questionnaire for authentic leadership (n=18), Multi-factor Leadership Questionnaire for transformational leadership (n=16) and transactional leadership (n=6), and Servant Organizatioanl Leadership Assessment for servant leadership (n=7). The individual factors related to leadership were organizational commitment, followed by job satisfaction and turnover intention. An organizational factor related to leadership was nursing performance outcome, followed by teamwork. Conclusion(s): The findings of this review suggest nursing leadership has critical role to improve both individual and organizational outcomes. Future studies are needed to conduct in various settings including community and to examine ralationship between nursing leadership and nursing quality related variables such as patient safety and turnover rates. In addition, development of measurment focused on unique nursing leadership traits would be recommended to accurateluy measure nursing leadership.

Universal model of bias-stress-induced instability in inkjet-printed carbon nanotube networks field-effect transistors

Jung, Haesun,Choi, Sungju,Jang, Jun Tae,Yoon, Jinsu,Lee, Juhee,Lee, Yongwoo,Rhee, Jihyun,Ahn, Geumho,Yu, Hye Ri,Kim, Dong Myong,Choi, Sung-Jin,Kim, Dae Hwan Elsevier 2018 Solid-state electronics Vol.140 No.-

<P><B>Abstract</B></P> <P>We propose a universal model for bias-stress (BS)-induced instability in the inkjet-printed carbon nanotube (CNT) networks used in field-effect transistors (FETs). By combining two experimental methods, i.e., a comparison between air and vacuum BS tests and interface trap extraction, BS instability is explained regardless of either the BS polarity or ambient condition, using a single platform constituted by four key factors: OH<SUP>−</SUP> adsorption/desorption followed by a change in carrier concentration, electron concentration in CNT channel corroborated with H<SUB>2</SUB>O/O<SUB>2</SUB> molecules in ambient, charge trapping/detrapping, and interface trap generation. Under negative BS (NBS), the negative threshold voltage shift (ΔV<SUB>T</SUB>) is dominated by OH<SUP>−</SUP> desorption, which is followed by hole trapping in the interface and/or gate insulator. Under positive BS (PBS), the positive ΔV<SUB>T</SUB> is dominated by OH<SUP>−</SUP> adsorption, which is followed by electron trapping in the interface and/or gate insulator. This instability is compensated by interface trap extraction; PBS instability is slightly more complicated than NBS instability. Furthermore, our model is verified using device simulation, which gives insights on how much each mechanism contributes to BS instability. Our result is potentially useful for the design of highly stable CNT-based flexible circuits in the Internet of Things wearable healthcare era.</P> <P><B>Highlights</B></P> <P> <UL> <LI> We proposed a bias stress induced instability model for CNT FETs in air/vacuum. </LI> <LI> The V<SUB>T</SUB> shift is composed of ad/desorption of OH<SUP>−</SUP>, electron/hole trapping, and D<SUB>it.</SUB> </LI> <LI> We verified our model by using device simulation. </LI> <LI> We provides insight into how much each mechanism contributes to instability. </LI> <LI> Our model is useful for stable flexible circuit designs in wearable healthcare era. </LI> </UL> </P>

항생제 선별 없이 alpha 1,3-Galactosyltransferase 유전자 넉아웃 기반 Membrane Cofactor Protein과 Thrombomodulin 유전자를 동시에 발현하는 세포라인 확립

이해선(Haesun Lee),박미령(Mi-Ryung Park),오건봉(Keon Bong Oh) 한국산학기술학회 2021 한국산학기술학회논문지 Vol.22 No.12

돼지와 영장류의 면역 차이로 이종이식 돼지 장기에서 보체 활성화, 염증반응 그리고 혈액 응고 등의 거부반응이 발생하며 수여 영장류를 사망에 이르게 한다. 인위적으로 Membrane cofactor protein(MCP) 및 Thrombomodulin(TBM)을 발현시키는 것은 이러한 거부반응으로부터 세포를 보호하고 영장류 생존기간을 연장한다고 보고되었으며 이전 연구에서 코돈 최적화 MCP(mMCP) 와 TBM cDNA를 활용한 발현 벡터를 구축한 바 있다. 본 연구의 목적은 항생제 저항성 유전자를 이용하지 않고 형질전환 세포 선별 시스템을 확립하며 alpha-1,3-Galactosyltransferase knock-out(GTKO)/mMCP/TBM 세포 라인을 개발하는 것이다. Nucleofection을 통해 mMCP/TBM 동시 발현 벡터를 도입한 GTKO 세포를 MCP에 대한 항체를 이용하여 MCP를 발현하는 세포만을 두 차례 분리하였다. 이를 통해 분리한 세포는 단일세포가 되도록 배양하였으며 유세포분석을 통한 MCP 발현 수준에 따라 단일 클론의 세포라인을 분류하여 선별하였다. 최종적으로 MCP 단백질 발현이 90% 이상인 GTKO/mMCP/TBM 유전자형 세포 라인 4종을 개발하였다. 선별된 클론들은 GTKO/mMCP/TBM 돼지 생산을 위한 핵이식 공여세포로 활용할 예정이다. 항생제 저항성 유전자를 사용하지 않는 선별 기술은 향후 의료용 원료 동물 생산에 효과적으로 활용될 수 있을 것으로 기대된다. The immunological barriers between pigs and primates result in complement activation, inflammation, and coagulation in xenografts, consequently leading to the recipient"s death. The intentional co-expression of membrane cofactor protein (MCP) and thrombomodulin (TBM) in pigs is known to contribute to the prolonged survival of xenografted recipient primates. In a previous study, we had constructed MCP/TBM co-expression vectors using codon-optimized MCP (mMCP) and TBM cDNA. The objectives of this study were to establish an antibiotic-free selection system and generate a pig cell line carrying mMCP and TBM expression vectors based on the alpha-1,3-galactosyltransferase knock-out (GTKO) genetic background. The mMCP/TBM co-expression vectors were introduced into GTKO pig fibroblasts. We isolated and selected only MCP-expressing cells using anti-MCP antibodies and cell separators twice. Additionally, we performed single-cell separation and clonal culture depending on the MCP expression level. Finally, we were able to obtain four GTKO/mMCP/TBM clones expressing MCP at levels of 90% and over. The selected clones will be used as donor cells for nuclear somatic cell transfer to produce GTKO/mMCP/TBM pigs for xenotransplantation. We believe that the antibiotic-free selection system established in this study could be useful to produce biomedical source animals without the antibiotic resistance gene.

단독주택지와 공동주택지의 교통사고 위험도 비교

이해선(LEE, Haesun),한상진(HAN, Sangjin),이혜진(LEE, Hyejin) 대한교통학회 2016 대한교통학회 학술대회지 Vol.75 No.-

아파트 등 공동주택단지는 주차면의 확충, 주차 공간의 지하화, 단지 내 차량 동선 처리 및 감속 등의 노력이 지속적으로 이루어지고 있으나 공공에서 개발하는 단독주택지는 이러한 노력이 부족하다. 이 때문에 공동주택지보다 단독주택지에서 불법주•정차, 보차혼용 등 보행안전을 위협하는 요소가 많이 관찰된다. 본 연구는 단독주택지와 공동주택지의 사고율을 비교하여, 주거유형별로 보행자 안전에 차이가 있는지 살펴보고자 하였다. 분석을 위해 보험회사에서 제공하는 교통사고 자료를 통해 일산신도시에서 발생한 최근 5년간(2011~2015년)의 사고 자료를 공동주택지와 단독주택지로 구분하여 수집하였으며, SPSS 19.0을 활용하여 T-test를 실시하였다. 분석을 통해 두 집단간 차이가 있음을 확인하였고, 이는 주거유형에 따라 사고 발생 확률과 심각도가 달라질 수 있다는 것을 시사한다. 이는 상대적으로 가로환경이 열악한 단독주택지의 보행자 안전을 확보하기 위해서는 도시를 조성하는 단계에서부터 사고 위험을 높일 수 있는 요소를 파악하고 개선해나가야 함을 시사한다.

모바일 클라우드 컴퓨팅 서비스를 위한 가상머신 위치선정 및 이주 알고리즘

변해선(Haesun Byun),이숙영(Sookyoung Lee),성윤영(Yoonyoung Sung),이미정(Meejeong Lee) 한국통신학회 2014 한국통신학회 학술대회논문집 Vol.2014 No.6

네트워크 자원의 효율적인 사용을 위한 ISP와 P2P 어플리케이션의 협력 방안

변해선 ( Haesun Byun ),이미정 ( Meejeong Lee ),이종화 ( Jonghwa Lee ),강신각 ( Singak Kang ) 한국정보처리학회 2009 한국정보처리학회 학술대회논문집 Vol.16 No.2

P4P는 P2P 통신을 함에 있어서 가장 적절한 피어를 선택하여 P2P 어플리케이션의 성능을 향상 시키면서 네트워크 자원의 효율적인 사용을 극대화하기 위해 제안된 프레임워크이다. 본 논문에서는 P4P 프레임워크에 기존 CoolStreaming 시스템을 적용하는 방안을 설명하고, ISP 개체와 P2P 시스템 개체의 협력 동작과정 및 그 개체들이 유지/관리하는 정보 등을 설명한다.

동유럽 국가와 유로존간의 경기 상호의존구조 분석

박해선(Park, Haesun),이철원(Lee, Chul-won),이상직(Lee, Sangjik) 한국외국어대학교 외국학종합연구센터 동유럽발칸연구소 2009 동유럽발칸연구 Vol.23 No.1

2004년이후 동유럽국가들은 EU에 가입하기 시작하면서 서유럽 중심의 유로존과 경제협력과 교역증대를 통해 경제성장 기반을 확보하려고 하고 있다. 이러한 노력은 궁극적으로 통화통합으로 이어질 가능성이 높다. 그러나 통화통합으로 경제교류가 증대되기 위해서는 유로존과 동유럽국가들의 경기동조화가 전제되어야 한다. 또한 유로존과 동유럽국가들 사이의 경기변동 정보의 전달구조를 파악한다면 이 지역의 경기변동 양상과 대응정책을 이해하는 데 도움이 된다. 본 연구에서는 동유럽 7개국(체코, 헝가리, 불가리아, 폴란드, 루마니아, 슬로바키아, 슬로베니아)과 유로존의 2000년 1월부터 2009년 5월까지 월별 산업생산지수를 데이터로 하여 경기동조화현상 존재여부 2경기변동 정보의 전달구조를 규명하였다. 동유럽국가들이 EU에 가입하기 시작한 2004년 5월을 기준으로 전후반기로 나누어 실증분석을 하였다. 잔차항의 상관계수분석을 보면 후반기에는 유로존과 동유럽국가 사이의 상관관계가 전반기보다 증가하여, 유로존과 동유럽국가들의 경기동조화현상이 커지는 것으로 추정되었다. 한편, 전후반기의 경기정보 전달구조는 차이가 있는 것으로 분석되었다. 전반기에는 유로존의 경기변동 정보가 헝가리상관체코에만 영향을 주고 나머지 동유럽국가들에는 전달되지 않았으나, 후반기에는 헝가리를 매개로 하여 모든 동유럽국가들에 영향을 끼치는 것으로 추정되었다. 이는 EU가입후 유로존과 동유럽국가들 사이에 경기변동의 동조성이 더 커진 것을 뒷받침하는 결과이나, EU가입전후로 동유럽국가와 유로존의 상관관계에 변화가 없다는 일부 기존의 연구결과와는 다른 결론이다. Several eastern European countries which have joined the EU are expected to adopt the euro in the near future. The monetary integration requires the synchronization of business cycles among the eastern European countries and the euro zone. Otherwise the adoption of the euro might cause negative effects on the eastern European countries. In this paper, we employ vector error correction model and DAG (Directed Acyclic Graphs) to analyze the synchronization and the structure of interdependence of business cycles in the area. We use monthly industrial production indices of 7 eastern European countries (Czech, Hungary, Bulgaria, Poland, Romania, Slovakia and Slovenia) and of the euro zone for our empirical analyses. Since the number of the EU member countries increased from 15 to 25 in May 2004, we divide the data into two periods: January 2000- April 2004 and May 2004 - May 2009. We found that in the first period the correlation coefficients between the eastern European countries and the euro zone are lower and, in some cases, negative. In the second period, however, the correlation coefficients are quite different. All of them are positive and their absolute values increase. DAG provides the structure of interdependence of business cycles among the countries we consider. In the first period, the information on the business cycles of the euro zone affects only Hungary and Czech while in the second period, the euro zone's business cycle information has effects on all of the eastern European countries through Hungary. In sum, we conclude that after eastern European countries joined the EU, economic integration among the eastern European countries and the euro zone has been established.

Production and Breeding of Transgenic Cloned Pigs Expressing Human CD73

Seung-Chan Lee,Haesun Lee,Keon Bong Oh,In-Sul Hwang,Hyeon Yang,Mi-Ryung Park,Sun-A Ock,,Jae-Seok Woo,Gi-Sun Im,Seongsoo Hwang 한국발생생물학회 2017 발생과 생식 Vol.21 No.2

One of the reasons to causing blood coagulation in the tissue of xenografted organs was known to incompatibility of the blood coagulation and anti-coagulation regulatory system between TG pigs and primates. Thus, overexpression of human CD73 (hCD73) in the pig endothelial cells is considered as a method to reduce coagulopathy after pig-to-non-humanprimate xenotransplantation. This study was performed to produce and breed transgenic pigs expressing hCD73 for the studies immune rejection responses and could provide a successful application of xenotransplantation. The transgenic cells were constructed an hCD73 expression vector under control porcine Icam2 promoter (pIcam2-hCD73) and established donor cell lines expressing hCD73. The numbers of transferred reconstructed embryos were 127 ± 18.9. The pregnancy and delivery rate of surrogates were 8/18 (44%) and 3/18 (16%). The total number of delivered cloned pigs were 10 (2 alive, 7 mummy, and 1 died after birth). Among them, three live hCD73-pigs were successfully delivered by Caesarean section, but one was dead after birth. The two hCD73 TG cloned pigs had normal reproductive ability. They mated with wild type (WT) MGH (Massachusetts General Hospital) female sows and produced totally 16 piglets. Among them, 5 piglets were identified as hCD73 TG pigs. In conclusion, we successfully generated the hCD73 transgenic cloned pigs and produced their litters by natural mating. It can be possible to use a mate for the production of multiple transgenic pigs such as α-1,3-galactosyltransferase knock-out /hCD46 for xenotransplantation.

Isolation and characterization of cultured chicken oviduct epithelial cells and in vitro validation of constructed ovalbumin promoter in these cells

Yang, Hyeon,Lee, Bo Ram,Lee, Hwi-Cheul,Jung, Sun Keun,Kim, Ji-Youn,No, Jingu,Shanmugam, Sureshkumar,Jo, Yong Jin,Lee, Haesun,Hwang, Seongsoo,Byun, Sung June Asian Australasian Association of Animal Productio 2021 Animal Bioscience Vol.34 No.8

Objective: Transgenic hens hold a great promise to produce various valuable proteins. Through virus transduction into stage X embryo, the transgene expression under the control of constructed chicken ovalbumin promoters has been successfully achieved. However, a validation system that can evaluate differently developed ovalbumin promoters in in vitro, remains to be developed. Methods: In the present study, chicken oviduct epithelial cells (cOECs) were isolated from oviduct tissue and shortly cultured with keratinocyte complete medium supplemented with chicken serum. The isolated cells were characterized with immunofluorescence, western blot, and flow cytometry using oviduct-specific marker. Chicken mutated ovalbumin promoter (Mut-4.4-kb-pOV) was validated in these cells using luciferase reporter analysis. Results: The isolated cOECs revealed that the oviduct-specific marker, ovalbumin protein, was clearly detected by immunofluorescence, western blot, and flow cytometry analysis revealed that approximately 79.40% of the cells contained this protein. Also, luciferase reporter analysis showed that the constructed Mut-4.4-kb-pOV exhibited 7.1-fold (p<0.001) higher activity in the cOECs. Conclusion: Collectively, these results demonstrate the efficient isolation and characterization of cOECs and validate the activity of the constructed ovalbumin promoter in the cultured cOECs. The in vitro validation of the recombinant promoter activity in cOECs can facilitate the production of efficient transgenic chickens for potential use as bioreactors.