• Title/Summary/Keyword: ThMB

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Optimization of Human Embryonic Stem Cells into Differentiation of Dopaminergic Neurons in Vitro: II. Genetically Modified Human Embryonic Stem Cells Treated with RA/AA or b-FGF

  • 신현아;김은영;이영재;이금실;조황윤;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.75-75
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    • 2003
  • Since the establishment of embryonic stem cell, pluripotency of the cells was known to allow differentiation of the cells into various cell types consisting whole body. Several protocols have been developed to induce expression of specific genes.. However, no precise protocol that will generate a single type of the cells from stem cells has been reported. In order to produce cells suitable for transplantion into brain of PD animal model, which arouse due to a progressive degeneration of dopaminergic neurons in midbrain, human embryonic stem cell (hESC, MB03) was transfected with cDNAs cording for tyrosine hydroxylase (TH). Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by the two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA/ascorbic acid (AA), embryoid bodies (EB, for 4days) derived from hES cells were exposed to RA (10$^{-6}$ M)/AA (50 mM) for 4 days, and were allowed to differentiate in N2 medium for 7, 14, 21, or 28 days. Exp. II) When bFGF was used, neuronal precursor cells were selected for 8 days in N2 medium after EB formation. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14, 21 or 28 days. By indirect immunocytochemical studies, proportion of cells expressing NF200 increased rapidly from 20% at 7 days to 70 % at 28 days in RA/AA-treated group, while those cells expressing NF160 decreased from 80% at 7 days to 10% at 28 days upon differentiation in N2 medium. However, in differentiation by RA/AA treatment system, there was a significant increase in proportion of neuron maturity (73%) at day 14 after N2 medium. TH#2/MB03 cells expressing TH are >90% when matured at the absence of either bDNF or TGF-$\alpha$. These results suggested that TH#2/MB03 cells could be differentiated in vitro into mature neurons by RA/AA.

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In vitro Neural Cell Differentiation of Genetically Modified Human Embryonic Stem Cells Expressing Tyrosine Hydroxylase (Tyrosine Hydroxylase 유전자가 주입된 인간 배아줄기세포의 체외 신경세포 분화)

  • Shin, Hyun-Ah;Kim, Eun-Young;Lee, Keum-Sil;Cho, Hwang-Yoon;Kim, Yong-Sik;Lee, Won-Don;Park, Se-Pill;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.1
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    • pp.67-74
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    • 2004
  • Objective: This study was to examine in vitro neural cell differentiation pattern of the genetically modified human embryonic stem cells expressing tyrosine hydroxylase (TH). Materials and Methods: Human embryonic stem (hES, MB03) cell was transfected with cDNAs cording for TH. Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA, embryoid bodies (EB, for 4 days) derived from TH#2/MB03 cells were exposed to RA ($10^{-6}M$)/AA ($5{\times}10^{-2}mM$) for 4 days, and were allowed to differentiate in N2 medium for 7, 14 or 21 days. Exp. II) When b-FGF was used, neuronal precursor cells were expanded at the presence of b-FGF (10 ng/ml) for 6 days followed by a final differentiation in N2 medium for 7, 14 or 21 days. Neuron differentiation was examined by indirect immunocytochemistry using neuron markers (NF160 & NF200). Results: After 7 days in N2 medium, approximately 80% and 20% of the RA or b-FGF induced Th#2/MB03 cells were immunoreactive to anti-NF160 and anti-NF200 antibodies, respectively. As differentiation continued, NF200 in RA treated cells significantly increased to 73.0% on 14 days compared to that in b-FGF treated cells (53.0%, p<0.05), while the proportion of cells expressing NF160 was similarly decreased between two groups. However, throughout the differentiation, expression of TH was maintained ($\sim$90%). HPLC analyses indicated the increased levels of L-DOPA in RA treated genetically modified hES cells with longer differentiation time. Conclusion: These results suggested that a genetically modified hES cells (TH#2/MB03) could be efficiently differentiated in vitro into mature neurons by RA induction method.

Genetically Modified Human Embryonic Stem Cells Expressing Nurr1 and Their Differentiation into Tyrosine Hydroxylase Positive Cells In Vitro

  • Cho, Hwang-Yun;Lee, Chang-Hyun;Kim, Eun-Young;Lee, Won-Don;Park, Sepill;Lim, Jin-Ho
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.272-272
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    • 2004
  • The objective of this study is to test whether human embryonic stem cells expressing Nurr1 (Nurr1-transfected hES cells) could be expressed TH according to neuronal differentiation. As an effort to direct differentiation of hES (MB03 registered in NIH) cells to dopamine-producing neuronal cells, Nurr1 was transfected using conventional transfection protocol into MB03 cell and examined the expression of tyrosine hydroxylase (TH) after differentiation induced by retinoic acid (RA) and ascorbic acid (AA). (omitted)

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Genetically Modified Human Embryonic Stem Cells Expressing Nurr1 and Their Differentiation into Tyrosine Hydroxylase Positive Cells in vitro.

  • Cho, Hwang-Yoon;Lee, Chang-Hyun;Kil, Kwang-Soo;Yoon, Ji-Yeon;Shin, Hyun-Ah;Lee, Gun-Soup;Lee, Young-Jae;Kim, Eun-Young;Park, SePill;Lim, Jin-Ho
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.104-104
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    • 2003
  • As an effort to direct differentiation of human embryonic stem (hES, MB03) cells to dopamine-producing neuronal cells, Nurr1 was transfected using conventional transfection protocol into MB03 and examined the expression of tyrosine hydroylase (TH) after differentiation induced by retinoic acid (RA) and ascorbic acid (AA). Experimentally, cells were transfected with linearized Nurr1 cDNA in pcDNA3.1 (+)-hygovernight followed by selection in medium containing hygromycin-B (150 $\mu$/ml). Expression of Nurr1 mRNA was confirmed by RT-PCR and protein by immunocytochemistry in the drug resistant clones. In order to study the effect of Nurr1 protein on the differentiation pattern of ES cells, one of the positive clones (MBNr24) was allowed to form embryoid body (EB) for 2 days and were induced to differentiate for another 4 days using RA (1 $\mu M$) and AA (50 mM) (2-/4+ protocol) followed by selection in N2 medium for 10 or 20 days. After 10 days in N2 medium, cells immunoreactive to anti-GFAP, anti-TH, or anti-NF200 antibodies were 38.8%, 11%, and 20.5%, respectively. After 20 days in N2 medium, cells expressing GFAP, TH, or NF200 were 28%, 15% and 44.8%, respectively but approximately 9% of MB03 expressed TH protein when the cells were induced to differentiate using a similar prorocol, These results suggest that ectopic expression of Nurr1 enhances generation of TH+ cells as well as neuronal cells when hES cells were differentiated by 2-/4+ protocol.

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Minimum Bandwidth Clock Recovery Algorithm for 10 Gigabit Ethernet (10 Gigabit Ethernet을 위한 최소 대역폭 클럭리커버리 알고리즘)

  • 성충환;전경규;김환우;김대영
    • Proceedings of the IEEK Conference
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    • 2001.09a
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    • pp.911-914
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    • 2001
  • 본 논문에서는 10Gigabit Ethernet 물리계충 전송 기술로서 IEEE 802.3 Higher Speed Study Group (HSSG)에서 검토했던 방법으로 선로부호화 방법이 있는데 그 중에서 국내 연구진에 의해 제안된 최소 대역폭 선로부호 MB810을 사용하여 10Gigabit Ethernet에서의 clock recovery 가능성에 대해 알아 본다. MB810 code를 사용하면 기존의 통신 시스템에서 필요로하는 대역폭을 반만 사용하여 전송할 수 있기 때문에 대역 효율이 좋아지나 이전의 일반적인 square law 방법으로는 clock recovery가 어렵다. 본 논문에서는 4th power law 방법을 사용했을때의 이론적인 해석과 시뮬레이션 결과를 보인다.

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Fault Diagnosis of Nonlinear Systems Based on Dynamic Threshold Using Neural Network (신경회로망을 이용한 동적 문턱값에 의한 비선형 시스템의 고장진단)

  • Soh, Byung-Seok;Lee, In-Soo;Jeon, Gi-Joon
    • Journal of Institute of Control, Robotics and Systems
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    • v.6 no.11
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    • pp.968-973
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    • 2000
  • Fault diagnosis plays an important role in the performance and safe operation of many modern engineering plants. This paper investigates the problem of fault detection using neural networks in dynamic systems. A general framework for constructing a nonlinear fault detection scheme for nonlinear dynamic systems containing modeling uncertaintly is proposed. The main idea behind the proposed approach is to monitor the physical system with an off -line learning neural network and then to approximate the upper and lower thresholds of acceleration of the nominal system with the model-based threshold(ThMB) method, The performance of the proposed fault detection scheme is investigated through simulations of a pendulum with uncertainty.

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Inactivated Sendai Virus Strain Tianjin Induces Apoptosis in Human Breast Cancer MDA-MB-231 Cells

  • Chen, Jun;Han, Han;Chen, Min;Xu, Xiao-Zhu;Wang, Bin;Shi, Li-Ying
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.12
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    • pp.5023-5028
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    • 2014
  • Sendai virus strain Tianjin is a novel genotype. Here, we investigate the antitumor and proapoptotic effects of ultraviolet-inactivated Sendai virus strain Tianjin (UV-Tianjin) on human breast cancer MDA-MB-231 cells in vitro, as well as the involvement of the apoptotic pathway in the mechanism of UV-Tianjin-induced antitumor effects. MTT assays showed that treatment with UV-Tianjin dose-dependently inhibited the proliferation of MDA-MB-231 cells but not normal MCF 10A breast epithelium cells. Hoechst staining and flow cytometric analysis revealed that UV-Tianjin induced apoptosis of MDA-MB-231 cells in a dose-dependent manner. Moreover, UV-Tianjin treatment resulted in reduction in the mitochondria membrane potential (MMP) and release of cytochrome complex (cyt c) via regulation of Bax and Bcl-2, as well as activation of caspase-9, caspase-3, Fas, FasL and caspase-8 in MDA-MB-231 cells. In summary, our study suggests that UV-Tianjin exhibits anticancer activity in human breast cancer MDA-MB-231 cells through inducing apoptosis, which may involve both the endogenous mitochondrial and exogenous death receptor pathways.

Induction of Tyrosine Hydroxylase by Nurr-1 in hES Cells

  • An So-Yeon;Lee Yeong-Jae;Kim Eun-Yeong;Jo Hyeon-Jeong;Choe Gyeong-Hui;Park Se-Pil;Im Jin-Ho
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.85-85
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    • 2002
  • As an effort to direct differentiation of human embryonic stem cells (hES, MB03) to dopamine-producing neuronal cells, we expressed Nurr-l in hES and examined the expression of tyrosine hydroxylase (TH) after bFGF induction. To introduce Nurr-l, hES cells were maintained in humidified chamber with 5% CO₂ and 95% air in DMEM/Fl2 supplemented with FBS (10%), penicillin (100U/㎖), and streptomycin (100㎍/㎖). (omitted)

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MB-OFDM UWB modem SoC design (MB-OFDM 방식 UWB 모뎀의 SoC칩 설계)

  • Kim, Do-Hoon;Lee, Hyeon-Seok;Cho, Jin-Woong;Seo, Kyeung-Hak
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.34 no.8C
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    • pp.806-813
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    • 2009
  • This paper presents a modem chip design for high-speed wireless communications. Among the high-speed communication technologies, we design the UWB (Ultra-Wideband) modem SoC (System-on-Chip) Chip based on a MB-OFDM scheme which uses wide frequency band and gives low frequency interference to other communication services. The baseband system of the modem SoC chip is designed according to the standard document published by WiMedia. The SoC chip consists of FFT/IFFT (Fast Fourier Transform/Inverse Fast Fourier Transform), transmitter, receiver, symbol synchronizer, frequency offset estimator, Viterbi decoder, and other receiving parts. The chip is designed using 90nm CMOS (Complementary Metal-Oxide-Semiconductor) procedure. The chip size is about 5mm x 5mm and was fab-out in July 20th, 2009.

Inhibitory Effect of D-chiro-inositol on Both Growth and Recurrence of Breast Tumor from MDA-MB-231 Cancer Cells

  • Kim, Yoon-seob;Park, Ji-sung;Kim, Minji;Hwang, Bang Yeon;Lee, Chong-kil;Song, Sukgil
    • Natural Product Sciences
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    • v.23 no.1
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    • pp.35-39
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    • 2017
  • D-chiro-inositol (DCI) is a secondary messenger in insulin signal transduction. It is produced in vivo from myo-inositol via action of epimerase. In this study, we evaluated antitumor activity of DCI against human breast cancer both in vitro and in vivo. In order to determine the inhibitory effects of DCI on growth of human breast cancer cells (MDA-MB-231), two different assessment methods were implemented: MTT assay and mouse xenograft assay. MTT assay demonstrated downturn in cell proliferation by DCI treatment (1, 5, 10, 20 and 40 mM) groups by 18.3% (p < 0.05), 17.2% (p < 0.05), 17.5% (p < 0.05), 18.4% (p < 0.05), and 24.9% (p < 0.01), respectively. Also, inhibition of tumor growth was investigated in mouse xenograft model. DCI was administered orally at the dose of 500 mg/kg and 1000 mg/kg body weight to treat nude mouse for 45 consecutive days. On the 45th day, tumor growth of DCI (500 mg/kg and 1000 mg/kg) groups was suppressed by 22.1% and 67.6% as mean tumor volumes were $9313.8{\pm}474.1mm^3$ and $3879.1{\pm}1044.1mm^3$, respectively. Furthermore, breast cancer stem cell (CSC) phenotype ($CD44^+/C24^-$) was measured using flow cytometry. On the 46th day, CSC ratios of DCI (500 mg/kg) and co-treatment with doxorubicin (4 mg/kg) and DCI (500 mg/kg) group decreased by 24.7% and 53.9% (p < 0.01), respectively. Finally, from tumor recurrence assay, delay of 5 days in the co-treatment group compared to doxorubicin (4 mg/kg) alone group was observed. Based on these findings, we propose that DCI holds potential as an anti-cancer drug for treatment of breast cancer.