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Chromosome Aberrations in Porcine Embryo Produced by Nuclear Transfer with Somatic Cell

  • Ah, Ko-Seung;Jin, Song-Sang;Tae, Do-Jeong;Chung, Kil-Saeng;Lee, Hoon-Taek
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.73-73
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    • 2002
  • Nuclear transfer (NT) techniques have advanced in the last years, and cloned animals have been produced by using somatic cells in several species including pig. However, it is difficult that the nuclear transfer porcine embryos development to blastocyst stage overcoming the cell block in vitro. Abnormal segregation of chromosomes in nuclear transferred embryos on genome activation stage bring about embryo degeneration, abnormal blastocyst, delayed and low embryo development. Thus, we are evaluated that the correlations of the frequency of embryo developmental rates and chromosome aberration in NT and In viかo fertilization (IVF) derived embryo. We are used for ear-skin-fibroblast cell in NT. If only karyotyping of embryonic cells are chromosomally abnormal, they may difficultly remain undetected. Then, we evaluate the chromosome aberrations, fluorescent in situ hybridization (FISH) with porcine chromosome 1 submetacentric specific DNA probe were excuted. In normal diploid cell nucleus, two hybridization signal was detected. In contrast, abnormal cell figured one or three over signals. The developmental rates of NT and IVF embryos were 55% vs 63%, 32% vs 33% and 13% vs 17% in 2 cell, 8 cell and blastocyst, respectively. When looking at the types of chromosome aberration, the detection of aneuploidy at Day 3 on the embryo culture. The percentage of chromosome aneuploidy of NT and IVF at 4-cell stage 40.0%, 31.3%, respectively. This result indicate that chromosomal abnormalities are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT associated with lower implantation rate, increase abortion rate and production of abnormal fetuses.

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Chromosome Aberrations in Porcine Embryo Produced by Nuclear Transfer with Somatic Cell

  • K. S. Chung;Ko, S. A;S. J. Song;J. T. Do;Park, Y. S.;Lee, H. T.
    • Korean Journal of Animal Reproduction
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    • v.26 no.4
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    • pp.385-394
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    • 2002
  • This study was constructed the correlations of the embryonic developmental rates and the frequency of chromosome aberration using ear-skin-fibroblast cell in nuclear transfer (NT) derived embryos. Karyoplast-oocyte complexes were fused and activated simultaneously, then cultured for seven days to assess development. The developmental rates of NT and in vitro fertilization (IVF) embryos were 55.4% vs 63.5%, 31.7% vs 33% and 13.4% vs 16.8% in 2 cell, 8 cell and blastocyst, respectively. Firstly, the frequency of chromosome aberrations were evaluated using fluorescent in situ hybridization (FISH) technique with porcine chromosome 1 submetacentric specific probe. Chromosome aberration was detected at day 3 on the embryo culture, the percentages of chromosomal aneuploidy in NT and IVF embryos at 4-cell stage were 40%, 31.3%, respectively. Secondly, embryonic fragmentation was evaluated at 4-cell stage embryo. Frequency of embryonic fragmentations was in 51.3% of NT, 61.3% of IVF, 28.9% of parthenogenetic activation at 4-cell stage. The proportion of fragmentation in NT embryos was higher than activation embryos. This result indicates that chromosomal abnormalities and embryonic fragments are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT related with lower implantation rate, increased abortion rate and production of abnormal fetuses.

siRNAs Derived from Cymbidium Mosaic Virus and Odontoglossum Ringspot Virus Down-modulated the Expression Levels of Endogenous Genes in Phalaenopsis equestris

  • Lan, Han-hong;Wang, Cui-mei;Chen, Shuang-shuang;Zheng, Jian-ying
    • The Plant Pathology Journal
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    • v.35 no.5
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    • pp.508-520
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    • 2019
  • Interplay between Cymbidium mosaic virus (CymMV)/Odontoglossum ringspot virus (ORSV) and its host plant Phalaenopsis equestris remain largely unknown, which led to deficiency of effective measures to control disease of P. equestris caused by infecting viruses. In this study, for the first time, we characterized viral small interfering RNAs (vsiRNAs) profiles in P. equestris co-infected with CymMV and ORSV through small RNA sequencing technology. CymMV and ORSV small interfering RNAs (siRNAs) demonstrated several general and specific/new characteristics. vsiRNAs, with A/U bias at the first nucleotide, were predominantly 21-nt long and they were derived predominantly (90%) from viral positive-strand RNA. 21-nt siRNA duplexes with 0-nt overhangs were the most abundant 21-nt duplexes, followed by 2-nt overhangs and then 1-nt overhangs 21-nt duplexes in infected P. equestris. Continuous but heterogeneous distribution and secondary structures prediction implied that vsiRNAs originate predominantly by direct Dicer-like enzymes cleavage of imperfect duplexes in the most folded regions of the positive strand of both viruses RNA molecular. Furthermore, we totally predicted 54 target genes by vsiRNAs with psRNATarget server, including disease/stress response-related genes, RNA interference core components, cytoskeleton-related genes, photosynthesis or energy supply related genes. Gene Ontology classification showed that a majority of the predicted targets were related to cellular components and cellular processes and performed a certain function. All target genes were down-regulated with different degree by vsiRNAs as shown by real-time reverse transcription polymerase chain reaction. Taken together, CymMV and ORSV siRNAs played important roles in interplay with P. equestris by down modulating the expression levels of endogenous genes in host plant.

Effect of Osmolarity of Culture Medium on the Preimplantation Development of Porcine NT and IVF Embryos

  • Hwang, In-Sun;Park, Mi-Rung;Moon, Hyo-Jin;Shim, Joo-Hyun;Kim, Dong-Hoon;Yang, Byoung-Chul;Ko, Yeoung-Gyu;Yang, Boh-Suk;Cheong, Hee-Tae;Im, GI-Sun
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.91-96
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    • 2007
  • In vitro development of porcine embryo is affected by culture condition. One possible factor is osmolarity of culture medium. 1his study examined whether high osmolarity of culture medium at the early culture stage improves development of preimplantation porcine in vitro fertilization (IVF) and nuclear transfer (NT) embryos. NT and IVF embryos were divided into three groups and the basic medium was PZM-3 ($250{\sim}270$ mOsmol, control group). The control group of embryos was cultured in PZM-3 for whole culture period. Other two groups of embryos were cultured in a modified PZM-3 with 0.05 M sorbitol or 0.05 M sucrose ($300{\sim}320$ mOsmol, sorbitol or sucrose group) for the first 2 days, and then cultured in PZM-3 for further culture. NT embryos cultured in sucrose group showed a significantly higher developmental rate to the blastocyst stage with a decreased apoptosis rate compared to the sorbitol (p<0.05). For IVF, sucrose group showed a significantly increased the blastocyst formation rate with a decreased apoptosis rate compared to the control (p<0.05). This study represents that the high osmolarity in the early embryo culture stage can enhance the in vitro development of porcine NT and IVF embryos to the blastocyst stage with reduced apoptosis of cells.

Thermoelectric Properties of Skutterudite CoSb3 Prepared by Arc Melting (아크용해법으로 제조된 Skutterudite CoSb3의 열전특성)

  • Yu S.W.;Park J.B.;Cho K.W.;Jang K.W.;Ur S.C.;Lee J.I.;Kim I.H.
    • Korean Journal of Materials Research
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    • v.15 no.2
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    • pp.93-96
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    • 2005
  • The arc melting was employed to prepare undoped $CoSb_3$ compounds and their thermoelectric properties were investigated. Specimen annealed at $400^{\circ}C$ for 24 hrs showed sound microstructure. However, considerable voids and cracks were found after annealing at above $500^{\circ}C$. It seems to be attributed to the phase dissociation and thermal expansion due to phase transitions during annealing and cooling. Single phase $\delta-CoSb_3$ was successfully obtained by annealing at $400^{\circ}C$ for 24 hrs. In the case of increasing annealing temperature, phase decompositions occurred. Undoped $CoSb_3$ showed p-type conduction and intrinsic semiconducting behavior at all temperatures examined. Thermoelectric properties were remarkably improved by annealing and they were closely related to phase transitions.

The Effect of Additional Haptic Supplementation on Postural Control During Squat in Normal Adult (추가적인 햅틱적용이 정상 성인의 스쿼트 동안 자세조절에 미치는 영향)

  • Kim, Mi-Ju;Lee, Ho-Cheol;Park, Ji-Won
    • The Journal of Korean Physical Therapy
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    • v.24 no.2
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    • pp.134-142
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    • 2012
  • Purpose: This study examined the effect on postural control during the stimulation of haptic touch with fingertip on the stable surface at quiet standing posture, squat flexion stage, 60 degrees squat stage and squat extension stage. Methods: The postural sway was measured on the force platform, while 30 subjects were squatting, under three different haptic touch conditions (No Touch [NT], Light Touch [LT], Heavy Touch [HT]), above the touch pad in front of their body midline. Three different haptic touch conditions were divided into 1) NT condition; squatting as right index fingers held above the touch pad, 2) LT condition (<1N); squatting as the touch pad was in contact with right index fingers pulp with a pressure not exceeding 1N and 3) HT condition; squatting as subjects were allowed to use the touch pad for mechanical support by transmitting onto it with as much force, choosing with their index fingers. Results: There was significant decrease in LT, rather than that of NT (p<0.01), and in HT, rather than that of LT (p<0.01), as the results of the distance and velocity of center of pressure (COP) in mediolateral direction at quiet standing position. In anteroposterior direction, the distance and velocity of COP in LT and HT showed significant decrease, when compared to that of the data of NT (p<0.01). There was no significant difference between the 3 conditions (NT, LT, and HT), with respect to the distance and velocity of COP in mediolateral direction, during dynamic balance (squat flexion stage, squat extension stage) (p>0.05). In anteroposterior direction, the results of the distance and velocity of COP in HT showed significant decrease when compared to that of the data of NT (p<0.05). Conclusion: Light touch, during the task, decreased the postural sway at static balance. The results suggest that haptic touch should be applied, appropriately, because it varies the effects according to different conditions.

Fasudil Increases the Establishment of Somatic Cell Nuclear Transfer Embryonic Stem Cells in Mouse

  • So, Seongjun;Karagozlu, Mustafa Zafer;Lee, Yeonmi;Kang, Eunju
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.1
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    • pp.21-27
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    • 2020
  • Somatic cell nuclear transfer derived embryonic stem cells (NT-ESCs) have significant advantages in various fields such as genetics, embryology, stem cell science, and regenerative medicine. However, the poor establishment of NT-ESCs hinders various research. Here, we applied fasudil, a Rho-associated kinase (ROCK) inhibitor, to develop somatic cell nuclear transfer (SCNT) embryos and establish NT-ESCs. In the study, MII oocytes were isolated from female B6D2F1 mice and performed SCNT with mouse embryonic fibroblasts (MEFs). The reconstructed NT-oocytes were activated artificially, and cultured to blastocysts in KSOM supplemented with 10 μM fasudil. Further, the blastocysts were seeded on inactivated MEFs in embryonic stem cell medium supplemented with 10 μM fasudil. A total of 26% of embryos formed into blastocysts in the fasudil treated group, while this ratio was 44% in the fasudil free control group. On the other hand, 30% of blastocysts were established NT-ESCs after exposure of fasudil, which was significantly higher than the control group (10%). The results suggest that fasudil reduced blastocyst development after SCNT due to inhibition of 2 cell cleavage while improved the establishment of NT-ESCs through the anti-apoptotic pathway.

N-Terminal Pro-B-type Natriuretic Peptide Is Useful to Predict Cardiac Complications Following Lung Resection Surgery

  • Lee, Chang-Young;Bae, Mi-Kyung;Lee, Jin-Gu;Kim, Kwan-Wook;Park, In-Kyu;Chung, Kyung-Young
    • Journal of Chest Surgery
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    • v.44 no.1
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    • pp.44-50
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    • 2011
  • Background: Cardiovascular complications are major causes of morbidity and mortality following non-cardiac thoracic operations. Recent studies have demonstrated that elevation of N-Terminal Pro-B-type natriuretic peptide (NT-proBNP) levels can predict cardiac complications following non-cardiac major surgery as well as cardiac surgery. However, there is little information on the correlation between lung resection surgery and NT-proBNP levels. We evaluated the role of NT-proBNP as a potential marker for the risk stratification of cardiac complications following lung resection surgery. Material and Methods: Prospectively collected data of 98 patients, who underwent elective lung resection from August 2007 to February 2008, were analyzed. Postoperative adverse cardiac events were categorized as myocardial injury, ECG evidence of ischemia or arrhythmia, heart failure, or cardiac death. Results: Postoperative cardiac complications were documented in 9 patients (9/98, 9.2%): Atrial fibrillation in 3, ECG-evidenced ischemia in 2 and heart failure in 4. Preoperative median NT-proBNP levels was significantly higher in patients who developed postoperative cardiac complications than in the rest (200.2 ng/L versus 45.0 ng/L, p=0.009). NT-proBNP levels predicted adverse cardiac events with an area under the receiver operating characteristic curve of 0.76 [95% confidence interval (CI) 0.545~0.988, p=0.01]. A preoperative NT-proBNP value of 160 ng/L was found to be the best cut-off value for detecting postoperative cardiac complication with a positive predictive value of 0.857 and a negative predictive value of 0.978. Other factors related to cardiac complications by univariate analysis were a higher American Society of Anesthesiologists grade, a higher NYHA functional class and a history of hypertension. In multivariate analysis, however, high preoperative NT-proBNP level (>160 ng/L) only remained significant. Conclusion: An elevated preoperative NT-proBNP level is identified as an independent predictor of cardiac complications following lung resection surgery.

Serial Survey on Group A beta-Hemolytic Streptococcal Carrier Rate and Serotyping in Elementary School Children in 1996~1998 (3년간(1996~1998) 초등학생의 A군 연쇄구균 보균율과 혈청학적 분류에 관한 연구)

  • Kim, Ji-Hyun;Kim, Ju-Ye;Kang, Hyeon-Ho;Cha, Sung-Ho;Lee, Young-Hee
    • Pediatric Infection and Vaccine
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    • v.7 no.1
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    • pp.143-151
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    • 2000
  • Purpose : The accuracy of bacteriologic diagnosis of beta-hemolytic streptococcal pharyngotonsillitis depends on the degree of carrier rate in that area where the throat swabs are obtained and the evaluation of serological T typing as an epidemiologic marker is important to understand epidemiology of streptococcal infection. The purpose of this study is to know the carrier rates of group A streptococcus in normal children form four different areas and to find out the epidemiologic characteristic in distribution of the serotypes for 3 years. Method : Throat swabs were obtained from the tonsillar fossa of normal school children in four different areas(Uljin, Seoul, Osan, Kunsan) from March to May 1996, in Uljin in April 1997, and in Uljin in April 1998. The samples were plated on a 5% sheep blood agar plate and incubated overnight at $37^{\circ}C$ before examination for the presence of beta-hemolytic colonies. All isolated beta-hemolytic streptococcus were grouped and serotyped by T agglutination. Results : The carrier rate of beta-hemolytic streptococci and group A streptococci in 1996 were 27.6%, 18.6% at Uljin; 16.4%, 2.7% at Seoul; 33.0%, 26.0% at Osan; 20.0%, 12.3% at Kunsan, respectively. Among 1,192 normal school children from 4 different areas, we obtained 179 strains of group A streptococci. Fifty two percent of the strains were typable by T agglutination in 1996. Common T-type in 1996 were NT, T1, T3, T2 at Uljin; T12, T25 at Seoul; NT, T6, T28 at Osan; T25, T4, NT, T5 at Kunsan, in decreasing order, respectively. At Uljin, T1, T3, T25 accounted for 69% of strains in 1996, T1, T12, T25 accounted for 70% in 1997, and T12, T4 accounted for 88% in 1998. Conclusion : Higher carrier rates were found in Uljin and Osan, where there are a lower population density with scanty of medical facilities compared with another areas. We supposed that low carrier rates is likely to be related to antibiotic abuse or some epidemiologic factor. The periodic and seasonal serotyping analysis is important in monitoring and understanding the epidemiologic patterns of group A streptococci.

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A Study on Nurturing Policy of IT-BT-NT Convergence Industry (IT-BT-NT 기술 융합에 따른 산업육성전략)

  • Chun, H.S.;Heo, P.S.
    • Electronics and Telecommunications Trends
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    • v.21 no.2 s.98
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    • pp.15-25
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    • 2006
  • 최근 IT, BT, NT 기술간 융합이 가속화되면서 새로운 형태의 기술과 서비스가 등장하고 있다. 미국, 일본, 유럽 등 선진국은 정부와 기업이 연대하여 기술간 융합을 고부가가치 창출에 필요한 첨단기술로 인식하고 융합기술 개발을 적극적으로 추진하고 있고, 우리 정부도 과학기술부, 산업자원부, 정보통신부를 중심으로 융합기술 발전계획을 수립하고 있다. 융합기술 개발을 통해 미래의 핵심기술을 확보하고, 컨버전스 시대의 조류에 대응하며, 새로운 성장동력을 창출하고, 건강하고 안전한 삶을 실현할 수 있다. 본고에서는 먼저 IT-BT-NT 융합기술의 분류, 특징 및 기대효과를 살펴보고, 산업육성전략 방안을 제시하고자 한다.