• 제목/요약/키워드: mMLC

검색결과 53건 처리시간 0.031초

Formation of Functional Cardiomyocytes Derived from Mouse Embryonic Stem Cells

  • 신현아;김은영;이영재;이금실;조황윤;박세필;임진호
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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    • pp.76-76
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    • 2003
  • Pluripotent embryonic stem cells can differentiate into beating cardiomyocytes with proper culture conditions and stimulants via embryo-like aggregates. We describe here the use of mouse embryonic stem (mES03) cells as a reproducible differentiation system for cardiomyocyte. mES03 cells growing in colonies were dissociated and allowed to re-aggregated in suspension [embryoid body (EB) formation〕. To induce cardiomyocytic differentiation, cells were exposed to 0.75% dimethyl sulfoxide (DMSO) during EB formation for 4 days and then another 4 days without DMSO (4+/4-). Thus treated EB was plated onto gelatin-coated dishes for differentiation. Spontaneously contracting colonies which appeared in approximately 4~5 days upon differentiation were mechanically dissected, enzymatically dispersed, plated onto coverslips, and then incubated for another 48~72 hrs. By RT-PCR, robust expression of cardiac myosin heavy chain $\alpha$, cardiac muscle heavy polypeptide 7 $\beta$($\beta$-MHC), cardiac transcription factor GATA4, and skeletal muscle-specific $\alpha$$_1$-subunit of the L-type calcium channel ($\alpha$$_1$CaC $h_{sm}$ ) were detected as early as 8 days after EB formation, but message of cardiac muscle-specific $\alpha$$_1$-subunit of the L-type calcium channel ($\alpha$$_1$CaCh) were reveled at a low level. In contrast, expression of myosin light chain (MLC-2V) and atrial natriuretic factor (ANF) were not detected during EB formation for 8 days. However, a strong expression of the atrial-specific ANF gene was expressed from day 8 onward, which were remained constant in EB. (cardiac specialization and terminal differentiation stage). Electrophysiological examination of spontaneously contracting cells showed ventricle-like action potential 17 days after the EB formation. This study indicates that mES03 cell-derived cardiomyocytes via 4+/4- protocol displayed biochemical and electrophysiological properties of subpopulation of cardiomyocytes.

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Activation of Small GTPases RhoA and Rac1 Is Required for Avian Reovirus p10-induced Syncytium Formation

  • Liu, Hung-Jen;Lin, Ping-Yuan;Wang, Ling-Rung;Hsu, Hsue-Yin;Liao, Ming-Huei;Shih, Wen-Ling
    • Molecules and Cells
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    • 제26권4호
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    • pp.396-403
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    • 2008
  • The first ORF of the ARV S1133 S1 segment encodes the nonstructural protein p10, which is responsible for the induction of cell syncytium formation. However, p10-dependent signaling during syncytium formation is fully unknown. Here, we show that dominant negative RhoA, Rho inhibitor C3 exoenzyme, ROCK/Rho-kinase inhibitor Y-27632 and Rac1 inhibitor NSC23766 inhibit p10-mediated cell fusion. p10 over-expression is concomitant with activation and membrane translocation of RhoA and Rac1, but not cdc42. RhoA and Rac1 downstream events, including JNK phosphorylation and transcription factor AP-1 and $NF-{\kappa}B$ activation, as well as MLC expression and phosphorylation are simultaneously activated by p10. p10 point mutant T13M possessed 20% fusion-inducing ability and four p10 fusion-deficient mutants V15M, V19M, C21S and L32A reduced or lost their ability to activate RhoA and Rac1 signaling. We conclude that p10-mediated syncytium formation proceeds by utilizing RhoA and Rac1-dependent signaling.

Melatonin inhibits the Migration of Colon Cancer RKO cells by Down-regulating Myosin Light Chain Kinase Expression through Cross-talk with p38 MAPK

  • Zou, Duo-Bing;Wei, Xiao;Hu, Ruo-Lei;Yang, Xiao-Ping;Zuo, Li;Zhang, Su-Mei;Zhu, Hua-Qing;Zhou, Qing;Gui, Shu-Yu;Wang, Yuan
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권14호
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    • pp.5835-5842
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    • 2015
  • Background: Melatonin, which is mainly produced by the pineal gland, has a good inhibitory effect on cell growth of multiple cancer types. However, the underlying molecular mechanisms of anti-tumor activity for colon cancer have not been fully elucidated. In this study, we investigated the effects of melatonin on migration in human colon cancer RKO cells and the potential molecular mechanisms. Materials and Methods: The viability of RKO cells was investigated by MTT assay after treatment with melatonin, SB203580 (p38 inhibitor) and phorbol 12-myristate 13-acetate (PMA, MAPK activator) alone or in combination for 48h. The effects of melatonin, and ML-7, a selective inhibitor of myosin light chain kinase (MLCK), and SB203580, and PMA on the migration of RKO cells were analyzed by in vitro scratch-wound assay. The relative mRNA levels of MLCK was assessed by real-time quantitative RT-PCR. Western blotting analysis was performed to examine the expression of MLCK, phosphorylation of myosin light chain (pMLC) and p38 (pp38). Results: The proliferation and migration of human colon cancer RKO cells were inhibited significantly after treatment with melatonin. The expression levels of MLCK and phosphorylation of MLC of RKO cells were reduced, and real-time quantitative RT-PCR showed that melatonin had significant effects on suppressing the expression of MLCK. Furthermore, the phosphorylation level of p38, which showed the same trend, was also reduced when cells were treated by melatonin. In addition, ML-7 (25umol/l) could down-regulate the phosphorylation of p38. Conclusions: Melatonin could inhibit the proliferation and migration of RKO cells, and further experiments confirmed that p38 MAPK plays an important role in regulating melatonin-induced migration inhibition through down-regulating the expression and activity of MLCK.

멀티레벨 셀을 가지는 PoRAM의 센싱 기법 (A Sensing Method of PoRAM with Multilevel Cell)

  • 이종훈;김정하;이상선
    • 대한전자공학회논문지SD
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    • 제47권12호
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    • pp.1-7
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    • 2010
  • 본 논문은 멀티레벨을 갖는 PoRAM 셀의 데이터를 센싱하는 기법에 관하여 제안하였다. PoRAM은 유기물질을 사용한 단위 셀의 상,하단 전극에 전압을 가했을 때 나타나는 저항 상태의 변화로 셀 데이터를 구분하는 메모리 소자이다. 특히 한 셀당 최대 4 레벨의 안정된 저항 값을 가지므로 멀티레벨 셀로 활용이 가능하다. 따라서 멀티레벨의 센싱을 위해 어드레스 디코딩 방법, 센스 앰플리파이어, 이를 위한 제어 신호 등을 새롭게 제안하였다. 센스 앰플리파이어는 셀에 흐르는 전류를 입력 값으로 받아 설정된 기준 전류($I_{REF}$)와 비교하는 전류 비교기를 기본으로 구성되며 전류를 증폭하기 위해 낮은 입력 임피던스를 갖도록 설계되었다. 제안된 기법에 의해 설계된 회로는 $0.13{\mu}m$ CMOS 공정 라이브러리를 사용하여 설계되었고, 이를 사용함으로써 단위 셀에 흐르는 서로 다른 4 가지 전류 값이 각각 데이터 "00", "01", "10", "11"으로 정확히 센싱 되는 것을 검증하였다.

요철 표면 팬텀을 이용한 치료계획시스템 선량계산 정확성 평가 (Accuracy Evaluation of Treatment Planning System Using Irregular-surface Water Phantom)

  • 정동혁;김기환;이강규;문성록;김진기;신교철;오영기;김정기;조문준;김준상
    • 한국의학물리학회지:의학물리
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    • 제19권2호
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    • pp.131-138
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    • 2008
  • 본 연구에서는 방사선치료에 사용되고 있는 치료계획시스템 중 AAA와 PBC (Batho, M-Batho, E-TAR)의 알고리즘이 내장된 Eclipse (Varian, USA)을 이용하여 요철 표면을 가진 팬텀에 대한 선량계산 정확성을 평가하였다. 측정 조사면은 $10{\times}10\;cm^2$$20{\times}20\;cm^2$, 그리고 각각에 대한 MLC 차폐 조사면을 대상으로 하였다. 팬텀 속 측정점은 SSD=90 cm에서 깊이 5, 10, 15 cm에서 전리함을 이용한 6MV X-선에 대한 측정값과 선량계산을 알고리즘 별로 수행하고 비교하였다. AAA알고리즘을 이용한 계산 값들은 평탄표면과 양각표면에서 ${\pm}1%$이내로 측정과 일치하였으며, 음각표면에서 단위 MU당 선량을 최대 -1.9%까지 낮게 평가하였다. PBC알고리즘을 이용한 계산 값들은 평탄표면과 양각표면에서 측정과 각각 최대 +1.7% 및 +4.1%의 차이로서 대체로 단위 MU당 선량을 높게 평가하였으며, 음각표면의 경우에 측정과 차이가 -3.1%에서 ${\pm}2.1%$ 범위를 보였다. 계산 값들을 AAPM과 IAEA 허용기준과 비교하고 또한 통계적으로 분석할 때, AAA의 결과가 측정과 가장 잘 일치하는 것으로 나타났으며, PBC의 경우에 Modified Batho 알고리즘이 비교적 측정과 잘 일치하는 것으로 평가되었다.

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방사선치료기기 정도관리를 위한 광도전체 센서 제작 및 평가 (The Evaluation and Fabrication of Photoconductor Sensor for Quality Assurance of Radiation Therapy Devices)

  • 강상식;노성진;정봉재;노시철;박지군
    • 한국방사선학회논문지
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    • 제10권8호
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    • pp.565-569
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    • 2016
  • 최근 다엽콜리메이터를 부착한 선형가속기를 이용한 방사선수술의 빈도가 점차 높아지고 있다. 이러한 정교한 방사선 수술은 소조사면 내에 고선량의 방사선이 집중적으로 조사되기 때문에 체계적이고 정확한 정도관리가 필수적이다. 본 연구는 PIB(Particle in Binder) 방식 중 침전법을 이용하여 $400{\mu}m$ 두께의 요오드화납($PbI_2$)과 요오드화수은($HgI_2$) 광도전체 센서 시편를 제작하였다. 제작된 시편의 전기적 특성은 암전류, 출력전류, 응답특성 및 선형성을 평가하였다. 평가 결과, $HgI_2$ 가 우수한 신호발생량과 선형성을 보였다. 끝으로, 두께에 따른 $HgI_2$ 센서의 신호반응 특성 결과, $400{\mu}m$ 두께에서 신호발생효율이 가장 높았고, ${\pm}2.5%$ 이내의 우수한 재현성을 보였다.

The Properties of Beam Intensity Scanner(BInS) in IMRT with Phantom for Three Dimensional Dose Verification

  • Young W. Vahc;Park, Kwangyl;Byung Y. Yi;Park, Kyung R.;Lee, Jong Y.;Ohyun Kwon;Park, Kwangyl;Kim, Keun M.
    • 한국의학물리학회:학술대회논문집
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    • 한국의학물리학회 2003년도 제27회 추계학술대회
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    • pp.64-64
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    • 2003
  • Objectives: Patient dose verification is clinically the most important parts in the treatment delivery of radiation therapy. The three dimensional(3D) reconstruction of dose distribution delivered to target volume helps to verify patient dose and determine the physical characteristics of beams used in intensity modulated radiation therapy(IMRT). We present Beam Intensity Scanner(BInS) system for the pre treatment dosimetric verification of two dimensional photon intensity. The BInS is a radiation detector with a custom made software for relative dose conversion of fluorescence signals from scintillator. Methods: This scintillator is fabricated by phosphor Gadolinium Oxysulphide and is used to produce fluorescence from the irradiation of 6MV photons on a Varian Clinac 21EX. The digitized fluoroscopic signals obtained by digital video camera will be processed by our custom made software to reproduce 3D relative dose distribution. For the intensity modulated beam(IMB), the BInS calculates absorbed dose in absolute beam fluence, which are used for the patient dose distribution. Results: Using BInS, we performed various measurements related to IMRT and found the followings: (1) The 3D dose profiles of the IMBs measured by the BInS demonstrate good agreement with radiographic film, pin type ionization chamber and Monte Carlo simulation. (2) The delivered beam intensity is altered by the mechanical and dosimetric properties of the collimating of dynamic and/or static MLC system. This is mostly due to leaf transmission, leaf penumbra, scattered photons from the round edges of leaves, and geometry of leaf. (3) The delivered dose depends on the operational detail of how to make multileaf opening. Conclusions: These phenomena result in a fluence distribution that can be substantially different from the initial and calculative intensity modulation and therefore, should be taken into account by the treatment planing for accurate dose calculations delivered to the target volume in IMRT.

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홍조류 모로우붉은실(Polysiphonia morrowii)의 추출물과 이로부터 분리된 브로모페놀계 화합물의 in vitro 항균·항스쿠티카충 활성 및 구조-활성 상관성 (In vitro Anti-bacterial and Anti-scuticociliate Activities of Extract and Bromophenols of the Marine Red Alga Polysiphonia morrowii with Structure-activity Relationships)

  • 강소영;이상윤;최준호;정성주
    • 한국수산과학회지
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    • 제47권1호
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    • pp.45-51
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    • 2014
  • Scuticociliates are regarded as serious pathogens in marine aquaculture worldwide. In Korea, they cause mass-mortalities in fish such as the commercially important olive flounder Paralichthys olivaceus. In particular, mixed infections of scuticociliates with pathogenic bacteria have been commonly reported. During efforts to identify natural marine-algae derived products that possess anti-bacterial and anti-scuticociliate properties, we found that an 80% methanolic extract of the red alga Polysiphonia morrowii Harvey exhibits both anti-scuticociliate activity against Miamiensis avidus, which is a major causative agent of scuticociliatosis, and anti-bacterial activities against fish pathogenic bacteria. Activity-guided fractionation and isolation of the 80% methanolic extract of P. morrowii yielded three bromophenols, which were identified as 3-bromo-4,5-dihydroxybenzyl methyl ether (1), 3-bromo-4,5-dihydroxybenzaldehyde (2) and urceolatol (3) based on spectroscopic analyses. 3-bromo-4,5-dihydroxybenzyl methyl ether (1) showed the highest anti-bacterial and anti-scuticociliate activities, with a minimal inhibitory concentration (MIC) of $62.5{\mu}g/mL$ (against Vibrio anguillarum) and minimal lethal concentration (MLC) of 62.5 ppm (in seawater). Investigations of the anti-bacterial and anti-scuticociliate activities of seventeen bromophenol derivatives, including the three isolated natural bromophenols, showed that the existence of an electron donating group or atom with a non-covalent electron pair at $C_4$ of the 2-bromophenol structure may be important in anti-scuticociliate activity. These findings suggest that the extract and bromophenol derivatives of P. morrowii may provide useful alternatives in aquaculture anti-scuticociliate therapies.

노발리스 ExacTrac system을 이용한 척추 정위 방사선수술 방법 평가 (Assessment of the Optic-guided Patient Positioning for Spinal Stereotactic Radiosurgery Using Novalis ExacTrac System)

  • 이동준;손문준;최광영;이기택;최찬영;황금철;황충진
    • 한국의학물리학회지:의학물리
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    • 제13권4호
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    • pp.218-223
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    • 2002
  • 두개외 병소 즉, 척추 등에 발생한 종양 혹은 혈관기형 등의 병소에 대해 침습적 수술이 불가한 경우 정위적 방사선 수술이 임상에서 시도되고 있다. 본 연구는 실험적으로 팬텀에 대해 척추 정위 방사선수술을 시행하고 결과 분석을 통하여 두개외 수술 목표점에 대한 방사선수술 위치 정확도를 평가하고자 한다. 이 실험을 위하여 팬텀을 특별히 고안하였으며 수술실에서 방사선 수술 목표점의 위치는 광학적 추적 장치를 이용하여 결정하였고 3 mm 미세다엽시준기(mMLC; mcro Multi-Leaf Collimator)를 사용하여 시술하였다. 팬텀의 목표점에 대한 방사선수술 오차는 $\pm$1 mm 이내였으며 분할방사선 수술 경우도 $\pm$1 mm 이내였다. 결과적으로 광학적 위치추적 장치를 이용한 두개외 목표점의 방사선 수술은 매우 정확하고 유용한 방법으로 판단된다.

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SPOT5영상과 현장조사자료를 융합한 대형산불지역의 피해강도 분석 (Analysis of Burn Severity in Large-fire Area Using SPOT5 Images and Field Survey Data)

  • 원명수;김경하;이상우
    • 한국농림기상학회지
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    • 제16권2호
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    • pp.114-124
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    • 2014
  • 본 연구는 2011년 100ha 이상의 대형산불 피해지인 울진과 영덕지역을 대상으로 하였으며 산불피해강도를 평가하기 위해 현장조사와 고해상도 위성영상자료 분석을 병행하였다. 위성영상자료 분석 시 산불피해지역의 피해강도를 가장 적절하게 평가할 수 있는 영상분류기법들을 비교 분석하여 최적의 피해강도 평가방법을 선정하였다. 대형산불 피해지역의 최적의 피해강도 평가기법으로는 현장조사에서 획득한 트레이닝 지역의 정보를 이용한 최대우도법을 적용하였을 때 가장 좋은 평가결과를 보였다. 산불피해강도의 정확도 검증 결과, 평균 전체정확도는 88.38%와 Kappa coefficient는 0.8147로 나타났다. 분류정확도는 최대우도법, NDVI, 최소거리법 순으로 나타났다. 산불피해강도 현장조사 결과와 위성영상자료에서 추출한 피해강도의 상관관계는 울진산불 피해지에서 r = -0.544, 영덕산불 피해지는 r = -0.616으로 나타났다.