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Transformation of Adult Mesenchymal Stem Cells into Cardiomyocytes with 5-azacytidine: Isolated from the Adipose Tissues of Rat (성체 백서의 지방조직에서 추출한 중간엽 줄기세포의 5-azacytidine을 이용한 심근세포 분화 유도)

  • Choe Ju-Won;Kim Yong-In;Oh Tae-Yun;Cho Dai-Yoon;Sohn Dong-Suep;Lee Tae-Jin
    • Journal of Chest Surgery
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    • v.39 no.7 s.264
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    • pp.511-519
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    • 2006
  • Background: Loss of cardiomyocytes in the myocardial infarction leads to regional contractile dysfunction, and necrotized cardiomyocytes in infracted ventricular tissues are progressively replaced by fibroblasts forming scar tissue. Although cardiomyoplasty, or implantation of ventricular assist device or artificial heart was tried in refractory heart failure, the cardiac transplantation was the only therapeutic modality because these other therapeutic strategies were not permanent. Cell transplantation is tried instead of cardiac transplantation, especially bone marrow is the most popular donated organ. But because bone marrow aspiration procedure is invasive and painful, and it had the fewer amounts of cellular population, the adipose tissue is recommended for harvesting of mesenchymal stem cells. Material and Method: After adipose tissues were extracted from abdominal subcutaneous adipose tissue and intra-abdominal adipose tissue individually, the cellular components were obtained by same method. These cellular components were tried to transformation with the various titers of 5-azacytidine to descript the appropriate concentration of 5-azacytidine and possibility of transformation ability of adipose tissue. Group 1 is abdominal subcutaneous adipose tissue and Group 2 is intra-abdominal adipose tissue-retroperitoneal adipose tissue and omentum. Cellular components were extracted by collagenase and $NH_4Cl$ et al, and these components were cultured by non-induction media - DMEM media containing 10% FBS and inducted by none, $3{\mu}mol/L,\;6{\mu}mol/L,\;and\;9{\mu}mol/L$ 5-azacytidine after the 1st and 2nd subculture. After 4 weeks incubation, tile cell blocks were made, immunostaining was done with the antibodies of CD34, heavy myosin chain, troponin T, and SMA. Result: Immunostaining of the transformed cells for troponin T was positive in the $6{\mu}mol/L\;&\;9{\mu}mol/L$ 5-azacytidine of Group 1 & 2, but CD34 and heavy myosin chain antibodies were negative and SMA antibody was positive in the $3{\mu}mol/L\;&\;6{\mu}mol/L$ 5-azacytidne of Group 2. Conclusion: These observations confirm that adult mesenchymal stem cells isolated from the abdominal subcutaneous adipose tissues and intra-abdominal adipose tissues can be chemically transformed into cardiomyocytes. This can potentially be a source of autologous cells for myocardial repair.

Variation in Cartenoid Pigment and lipids of the Arkshell, (Anadara broughtonii) according to the Environmental Factors of the Growing Area (서식환경요인에 따른 피조개육의 Carotenoid색소와 지질성분의 변화)

  • Ha, Bong-Senk;Kang, Dong-Soo;Kim, Yung-Gwan;Kim, Kui-Shik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.18 no.1
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    • pp.71-92
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    • 1989
  • The seasonal variations of environmental sea water, arid carotenoid and lipid composition of reddish muscle tissue of arkshell, Anadara broughtonii in Chungmu and $Y{\check{o}}su-area$ were investigated. In the sea water, pH value, salinity, electrical conductivity of Chungmu and $Y{\check{o}}su-area$ were similar tendency, but water temperature in annual average at Chungmu-area was about $2^{\circ}C$ higher than that of $Y{\check{o}}su-area$. The concentration of nitrite-nitrogen, ammonia-nitrogen, phosphate-phosphours at Chungmu-area were slightly higher than those at $Y{\check{o}}su-area$. Meanwhile the concentration of silicate-silicious at $Y{\check{o}}su-area$ was about 2 times higher than that of Chungmu-area. The bacterial density of the sea water was ranged from 3.6 to 93/100ml for coliform and 3.0 to 15/100ml for fecal colifrom at Chungmu-area, but the coliform was ranged 7.3 to 150/100ml and the fecal coliform was ranged 3.6 to 20/100ml at $Y{\check{o}}su-area$. Among the coliform, 8.9% Escherichia coli, 33.9% Citrobacter freundii, 41.1% Enterobacter aerogenes groups were classified and 16.1% was not identified. In the hemoglobin content of reddish muscle tissue of arkshell, one from Chungmu-area was ranged from 1.1 to 2.5 g /dl and one from $Y{\check{o}}su-area$ was ranged from 1.7 to 4.4 g /dl. In total carotenoid content, 0.80 to 1.28mg/100 g muscle was in Chungmu-area and 0.45 to 0.99mg/100 g muscle was in $Y{\check{o}}su-area$. Among the carotenoid compositions, pectenolone content was 63.0% in annual average from Chungmu area and 59.6% from $Y{\check{o}}su-area$, 6.5% from Chungmu area and 18.9% from $Y{\check{o}}su-area$ in ${\beta}-carotene$, 9.9% from Chungmu-area and 9.1% from Yosu-area in pectenoxanthin, 11.2% from Chungmu-area and 5.2% from $Y{\check{o}}su-area$ in diatoxanthin monoester were observed. In the seasonal variation for the major carotenoids, the content of pectenolone was continuously incresedbefore spawning period and showed the maximum in July, but decresed after spawning period. Diatoxanthin monoester was gradually incresed during all growing period from March to next January. And ${\beta}-carotene$ was decresed before spawning period and showed the maximum in July but incresed after spawning period. The major fatty acids of total lipid and fractionated lipid classes from total lipid namely neutral-, glyco- and phospholipid were $C_{16:0},\;C_{18:1},\;C_{22:6},\;C_{20:2},\;C_{18:2},\;C_{20:5}\;and\;C_{18:3}$ acid in the muscle tissue of arkshell. The content of neutral lipid showed about 2 times higher than polar lipid in the muscle tissue of arkshell from both Chungmu and, $Y{\check{o}}su-area$. The content of total and neutral lipid was decresed during spawning period. In the seasonal variation for fatty acid compositions, the content of polyenenoic acid in total and neutral lipid was decresed and saturated acid was incresed. On contrary, polyenoic acid in glyco and phospholipid was incresed and saturated acid was decresed during spawning period from July to September. The sterol composition in total lipid were mainly consisted of cholesterol, campesterol, brassicasterol and ${\beta}-sitosterol$. Especially, the rate of content in cholesterol showed decrese during spawning period butcampesterol showed increse from March to November.

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Effects of Recipient Oocytes and Electric Stimulation Condition on In Vitro Development of Cloned Embryos after Interspecies Nuclear Transfer with Caprine Somatic Cell (수핵난자와 전기적 융합조건이 산양의 이종간 복제수정란의 체외발달에 미치는 영향)

  • 이명열;박희성
    • Reproductive and Developmental Biology
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    • v.28 no.1
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    • pp.21-27
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    • 2004
  • This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Recipient bovine and porcine oocytes were obtained from slaughterhouse and were matured in vitro according to established protocols. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS and primary fibroblast cultures were established in TCM-199 with 10% FBS. The matured oocytes were dipped in D-PBS plus 10% FBS + 7.5 $\mu$ g/ml cytochalasin B and 0.05M sucrose. Enucleation were accomplished by aspirating the first polar body and partial cytoplasm which containing metaphase II chromosomes using a micropipette with an out diameter of 20∼30 $\mu$m. A Single donor cell was individually transferred into the perivitelline space of each enucleated oocyte. The reconstructed oocytes were electric fusion with 0.3M mannitol fusion medium. After the electrofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. And porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6 ∼8 day at $39^{\circ}C, 5% CO_2 $in air. Interspecies nuclear transfer by recipient bovine oocytes were fused with electric length 1.95 kv/cm and 2.10 kv/cm. There was no significant difference between two electric length in fusion rate(47.7 and 44.6%) and in cleavage rate(41.9 and 54.5%). Using electric length 1.95 kv/cm and 2.10 kv/cm in caprine-porcine NT oocytes, there was also no significant difference between two treatments in fusion rate(51.3 and 46.1%) and in cleavage rate(75.0 and 84.9%). The caprine-bovine NT oocytes fusion rate was lower(P<0.05) in 1 pulse for 60 $\mu$sec(19.3%), than those from 1 pulse for 30 $\mu$sec(50.8%) and 2 pulse for 30 $\mu$sec(31.0%). The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(53.3%) and 2 pulse for 30 $\mu$sec(50.0%), than in 1 pulse for 60 $\mu$sec(18.2%). The caprine-porcine NT oocytes fusion rate was 48.1% in 1 pulse for 30 $\mu$sec, 45.2% in 2 pulse for 30 $\mu$sec and 48.6% in 1 pulse for 60 $\mu$sec. The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(78.4%) and 1 pulse for 60 $\mu$sec(79.4%), than in 2 pulse for 30 $\mu$sec(53.6%). In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer and 30.6% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P<0.05) in interspecies nuclear transfer(5.1%) than parthenotes(37.4%).

A Study on the Daesoon Cosmology of the Correlative Relation between Mugeuk and Taegeuk (무극과 태극 상관연동의 대순우주론 연구)

  • Kim, Yong-hwan
    • Journal of the Daesoon Academy of Sciences
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    • v.33
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    • pp.31-62
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    • 2019
  • The purpose of this article is to study on the Daesoon Cosmology of the Correlative Relation between Mugeuk and Taegeuk. Daesoon cosmology is a cosmology based on the juxtaposition between the Gucheon Sangje and the world. In this article, I would like to say that this theory in Daesoon Thought was developed in three stages: the phase of the Mugeuk Transcendence of Gucheon Sangje, the stage of the Taegeuk Immanence, and the phase of the Grand Opening of the Later World between Mugeuk and Taegeuk as a correlative gentle reign. First of all, the phase of the Mugeuk Transcendence of Gucheon Sangje has been revealed as a yin-yang relationship. The stage of the Taegeuk Immanence represents the togetherness of harmony and co-prosperity between yin and yang, and the phase of the Grand Opening of the Later World between Mukeuk and Taegeuk refers to the unshakable accomplishment of its character and energy. It will be said that this is due to the practical mechanism in the correct balance of yin-yang making a four stage cycle of birth, growth, harvest, and storage. In addition, the Daesoon stage of the settlement of yin and yang is revealed as a change in the growth of all things and the formation of the inner circle. The mental growth reveals the characteristics of everything in the world, each trying to shine at the height of their own respective life as they grow up energetically. The dominant culture of cerebral communion renders a soft and elegant mood and combines yin and yang to elevate the heavenly and earthly period through transcendental change into sympathetic understanding. The stage of the Grand Opening of the Later World between Mugeuk and Taegeuk is one of the earliest days of the lunar month and also the inner circle of Taegeuk. It is in line with Ken Wilbur's integrated model as a step to the true degrees to develop into a world with brightened degrees. It is a beautiful and peaceful scene where celestial maidens play music, the firewood burns, and the scholars command thunder and lightning playfully. Human beings achieve a state of happiness as a free beings who lives as gods upon the earth. This is the world of theGrand Opening of the Later World between Mugeuk and Taegeuk. Daesoon Thought was succeeded by Dojeon in 1958, when Dojeon emerged as the successor in the lineage of religious orthodoxy and was assigned the task of handling Dao in its entirety. In addition, Daesoon is a circle and represents freedom and commonly shared happiness among the populous. Cosmology in the Daesoon Thought will enable us to understand deep dimensions and the identity of members as individuals within an inner circle of correlation between transcendence and immanence. This present study tries to analyze the public effects philologically and also the mutual correlation by utilizing the truthfulness of literature and rational interpretation. The outlook for the future in Daesoon Thought also leads to the one-way communication of Daesoon as a circle.