• Title/Summary/Keyword: 2-cell block

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Effects of Cell Cycle Regulators on the Cell Cycle Synchronization of Porcine induced Pluripotent Stem Cells

  • Kwon, Dae-Jin;Hwang, In-Sul;Kwak, Tae-Uk;Yang, Hyeon;Park, Mi-Ryung;Ock, Sun-A;Oh, Keon Bong;Woo, Jae-Seok;Im, Gi-Sun;Hwang, Seongsoo
    • Development and Reproduction
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    • v.21 no.1
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    • pp.47-54
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    • 2017
  • Unlike mouse results, cloning efficiency of nuclear transfer from porcine induced pluripotent stem cells (piPSCs) is very low. The present study was performed to investigate the effect of cell cycle inhibitors on the cell cycle synchronization of piPSCs. piPSCs were generated using combination of six human transcriptional factors under stem cell culture condition. To examine the efficiency of cell cycle synchronization, piPSCs were cultured on a matrigel coated plate with stem cell media and they were treated with staurosporine (STA, 20 nM), daidzein (DAI, $100{\mu}M$), roscovitine (ROSC, $10{\mu}M$), or olomoucine (OLO, $200{\mu}M$) for 12 h. Flow Cytometry (FACs) data showed that piPSCs in control were in G1 ($37.5{\pm}0.2%$), S ($34.0{\pm}0.6%$) and G2/M ($28.5{\pm}0.4%$). The proportion of cells at G1 in DAI group was significantly higher than that in control, while STA, ROSC and OLO treatments could not block the cell cycle of piPSCs. Both of viability and apoptosis were affected by STA and ROSC treatment, but there were no significantly differences between control and DAI groups. Real-Time qPCR and FACs results revealed that DAI treatment did not affect the expression of pluripotent gene, Oct4. In case of OLO, it did not affect both of viability and apoptosis, but Oct4 expression was significantly decreased. Our results suggest that DAI could be used for synchronizing piPSCs at G1 stage and has any deleterious effect on survival and pluripotency sustaining of piPSCs.

Coolant Leak Effect on Polymer Electrolyte Membrane Fuel Cell (고분자전해질연료전지의 냉각수 누설에 대한 연구)

  • Song, Hyun-Do;Kang, Jung-Tak;Kim, Jun-Bom
    • Journal of the Korean Electrochemical Society
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    • v.10 no.4
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    • pp.301-305
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    • 2007
  • The performance of polymer electrolyte membrane fuel cell could be decreased due to coolant leaked from connection part. Micro pump was used to put small amount of coolant and investigate the effect on fuel cell. The stoichiometric ratio of hydrogen/air was 1.5/2.0, both side of gas was fully humidified, and current density of $400mA/cm^2$ was used as standard condition in this experiment. Constant current method was used to check performance recovery from coolant effect in 3 cell stack. The performance was recovered when coolant was injected in cathode side. On the other hand, the performance was not recovered when coolant was injected in anode side. Ethylene glycol could be converted to CO in oxidation process and cause poisoning effect on platinum catalyst or be adhered on GDL and cause gas diffusion block effect resulting performance decrease. Water with nitrogen gas was supplied in anode side to check performance recovery. Polarization curve, cyclic voltammetry, electrochemical impedance spectroscopy was used to check performance, and gas chromatography was used to check coolant concentration. Constant current method was not enough in full recovery of performance. However, water injection method was proved good method in full recovery of performance.

Fine Needle Aspiration Cytology of Hepatoblastoma -Report of Two Cases- (간아세포종의 세침흡인 세포학적 소견 -2예 증례보고-)

  • Park, Young-Nyun;Lee, Kwang-Gil;Park, Chan-Il
    • The Korean Journal of Cytopathology
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    • v.1 no.1
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    • pp.98-102
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    • 1990
  • Hepatoblastoma (HB) is a rare embryonic malignant tumor of the liver. Most morphological studies on HB have limited to the histological characteristics and only 3 cases of HB have been described in the cytology literature. We present 2 cases of HB occurring in children aged 1 year and 3 years, respectively. The distinctive cytologic features of fine needle aspiration of HB were clusters of tumor cells showing acinar and trabecular pattern, smaller tumor cells with a high nuclear-cytopalsmic ratio and hyperchromatic nuclei having prominent nucleoli, and the presence of extramedullary hematopoiesis and osteoid material. These features were also found in the cell block and the biopsy specimen, and appeared very useful in the differentiation of HB from hepatocellular carcinoma.

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Fabrication of Microcapsules Encapsulating Fluorescent Nanoparticles and Visualization of Their Inclusion (형광 나노입자를 수용하는 마이크로캡슐의 제작 및 수용 가시화)

  • Kim, Eun-Young;Kim, Hyoung-Hoon;Go, Jeung-Sang
    • Journal of the Korean Society of Visualization
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    • v.9 no.2
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    • pp.16-20
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    • 2011
  • This paper presents a fabrication method of microcapsules encapsulating fluorescent nanoparticles sensitive to an organic liquid, which is potentially applicable to the encapsulation of protein, cell and drug. It uses the supra-molecular self-assembly of a block copolymer at the interface of the stable and controllable droplets of water suspended with fluorescent nanoparticles and the polymer solved organic. The size and uniformity of the microcapsules were examined for the various polymer concentrations by using SEM image analysis. The maximum standard deviation of the produced microcapsules of less than 3.5% was obtained from the microcapsules produced from the same conditions. The inclusion of fluorescent nanoparticles was visualized in the fluorescence microscope and by using TEM image. It is shown that this fabrication method can provide the uniform size microcapsules with a higher inclusion.

Effect of Genistein, a Tyrosine Kinase Inhibitor, on the Cloned Rat Brain Potassium Channel Kv1.5

  • Choi, Bok-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.10 no.5
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    • pp.243-249
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    • 2006
  • The effect of genistein, widely used as a specific tyrosine kinase inhibitor, on rat brain Kv1.5 channels which were stably expressed in Chinese hamster ovary cells was investigated using the whole-cell patch-clamp technique. Genistein inhibited Kv1.5 currents at +50 mV in a concentration-dependent manner, with an $IC_{50}$ of $54.7{\pm}8.2\;{\mu}M$ and a Hill coefficient of $1.1{\pm}0.2$. Pretreatment of Kv1.5 with protein tyrosine kinase inhibitors ($10\;{\mu}M$ lavendustin A and $100\;{\mu}M$ AG1296) and a tyrosine phosphatase inhibitor ($500\;{\mu}M$ sodium orthovanadate) did not block the inhibitory effect of genistein. The inhibition of Kv1.5 by genistein showed voltage-independence over the full activation voltage range positive to 0 mV. The activation (at +50 mV) kinetics was significantly delayed by genistein: time constant for an activation of $1.4{\pm}0.2$ msec under control conditions and $10.0{\pm}1.5$ msec in the presence of $60\;{\mu}M$ genistein. Genistein also slowed the deactivation of the tail currents, resulting in a crossover phenomenon: a time constant of $11.4{\pm}1.3$ msec and $40.0{\pm}4.2$ msec under control conditions and in the presence of $60\;{\mu}M$ genistein, respectively. Inhibition was reversed by the application of repetitive depolarizing pulses, especially during the early part of the activating pulse. These results suggest that genistein directly inhibits Kv1.5 channels, independent of phosphotyrosine-signaling pathway.

Open channel block of Kv1.4 potassium channels by aripiprazole

  • Park, Jeaneun;Cho, Kwang-Hyun;Lee, Hong Joon;Choi, Jin-Sung;Rhie, Duck-Joo
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.6
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    • pp.545-553
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    • 2020
  • Aripiprazole is a quinolinone derivative approved as an atypical antipsychotic drug for the treatment of schizophrenia and bipolar disorder. It acts as with partial agonist activities at the dopamine D2 receptors. Although it is known to be relatively safe for patients with cardiac ailments, less is known about the effect of aripiprazole on voltage-gated ion channels such as transient A-type K+ channels, which are important for the repolarization of cardiac and neuronal action potentials. Here, we investigated the effects of aripiprazole on Kv1.4 currents expressed in HEK293 cells using a whole-cell patch-clamp technique. Aripiprazole blocked Kv1.4 channels in a concentration-dependent manner with an IC50 value of 4.4 μM and a Hill coefficient of 2.5. Aripiprazole also accelerated the activation (time-to-peak) and inactivation kinetics. Aripiprazole induced a voltage-dependent (δ = 0.17) inhibition, which was use-dependent with successive pulses on Kv1.4 currents without altering the time course of recovery from inactivation. Dehydroaripiprazole, an active metabolite of aripiprazole, inhibited Kv1.4 with an IC50 value of 6.3 μM (p < 0.05 compared with aripiprazole) with a Hill coefficient of 2.0. Furthermore, aripiprazole inhibited Kv4.3 currents to a similar extent in a concentration-dependent manner with an IC50 value of 4.9 μM and a Hill coefficient of 2.3. Thus, our results indicate that aripiprazole blocked Kv1.4 by preferentially binding to the open state of the channels.

A Bacterial Metabolite, Compound K, Induces Programmed Necrosis in MCF-7 Cells via GSK3β

  • Kwak, Chae Won;Son, Young Min;Gu, Min Jeong;Kim, Girak;Lee, In Kyu;Kye, Yoon Chul;Kim, Han Wool;Song, Ki-Duk;Chu, Hyuk;Park, Byung-Chul;Lee, Hak-Kyo;Yang, Deok-Chun;Sprent, Jonathan;Yun, Cheol-Heui
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1170-1176
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    • 2015
  • Ginsenosides, the major active component of ginseng, are traditionally used to treat various diseases, including cancer, inflammation, and obesity. Among these, compound K (CK), an intestinal bacterial metabolite of the ginsenosides Rb1, Rb2, and Rc from Bacteroides JY-6, is reported to inhibit cancer cell growth by inducing cell-cycle arrest or cell death, including apoptosis and necrosis. However, the precise effect of CK on breast cancer cells remains unclear. MCF-7 cells were treated with CK ($0-70{\mu}M$) for 24 or 48 h. Cell proliferation and death were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry assays, respectively. Changes in downstream signaling molecules involved in cell death, including glycogen synthase kinase $3\beta$ ($GSK3\beta$), $GSK3\beta$, $\beta$-catenin, and cyclin D1, were analyzed by western blot assay. To block $GSK3\beta$ signaling, MCF-7 cells were pretreated with $GSK3\beta$ inhibitors 1 h prior to CK treatment. Cell death and the expression of $\beta$-catenin and cyclin D1 were then examined. CK dose- and time-dependently inhibited MCF-7 cell proliferation. Interestingly, CK induced programmed necrosis, but not apoptosis, via the $GSK3\beta$ signaling pathway in MCF-7 cells. CK inhibited $GSK3\beta$ phosphorylation, thereby suppressing the expression of $\beta$-catenin and cyclin D1. Our results suggest that CK induces programmed necrosis in MCF-7 breast cancer cells via the $GSK3\beta$ signaling pathway.

Differential Diagnosis between Small Cell Carcinoma and Adenocarcinoma of Lung in Fine Needle Aspiration Cytology (세침흡인 세포검사에서 폐의 소세포암종과 샘암종의 감별진단)

  • Choi, Young-Hee;Koh, Jae-Soo;Park, Sun-Hoo;Kim, Min-Suk;Cho, Soo-Youn;Kim, Jung-Soon;Ha, Hwa-Jung;Lee, Seung-Sook
    • The Korean Journal of Cytopathology
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    • v.17 no.2
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    • pp.120-125
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    • 2006
  • Distinguishing small cell carcinoma from other lung malignancies is of great clinico-therapeutic significance. Small cell carcinoma is an aggressive tumor with a tendency to metastasize early. Survival time if untreated is low but this tumor is highly responsive to chemotherapy. We have occasionally experienced difficulties in differentiation between adenocarcinoma and small cell carcinoma of the lung in fine needle aspiration cytology (FNAC). The aim of this study was to investigate the possibility of distinguishing small cell carcinoma from adenocarcinoma of the lung in FNAC. We evaluated cytomorphological features of FNAC specimens from 62 small cell carcinomas and 57 adenocarcinomas from the lung that were confirmed by biopsy and/or immunohistochemistry on cell block. Cytomorphological details of the two tumors were compared. Nuclear smearing and nearly absent cytoplasm were the most distinct findings in small cell carcinoma compared to adenocarcinoma (p<0.05). Necrotic background, architecture and chromatin pattern, nuclear molding and nucleoli were significantly different (p<0.05). Nuclear size, nuclear membrane nature and nuclear size variation however were not helpful in distinguishing the two tumors. Combining several features described above, small cell carcinoma can be properly differentiated from adenocarcinoma on FNAC. FNAC is proposed as a diagnostic tool of small cell carcinoma of the lung in the case of inaccessibility to biopsy, and so may allow the proper therapeutic strategies to be determined in such cases

Hardware Architecture of High Performance Cipher for Security of Digital Hologram (디지털 홀로그램의 보안을 위한 고성능 암호화기의 하드웨어 구조)

  • Seo, Young-Ho;Yoo, Ji-Sang;Kim, Dong-Wook
    • Journal of Broadcast Engineering
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    • v.17 no.2
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    • pp.374-387
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    • 2012
  • In this paper, we implement a new hardware for finding the significant coefficients of a digital hologram and ciphering them using discrete wavelet packet transform (DWPT). Discrete wavelet transform (DWT) and packetization of subbands is used, and the adopted ciphering technique can encrypt the subbands with various robustness based on the level of the wavelet transform and the threshold of subband energy. The hologram encryption consists of two parts; the first is to process DWPT, and the second is to encrypt the coefficients. We propose a lifting based hardware architecture for fast DWPT and block ciphering system with multi-mode for the various types of encryption. The unit cell which calculates the repeated arithmetic with the same structure is proposed and then it is expanded to the lifting kernel hardware. The block ciphering system is configured with three block cipher, AES, SEED and 3DES and encrypt and decrypt data with minimal latency time(minimum 128 clocks, maximum 256 clock) in real time. The information of a digital hologram can be hided by encrypting 0.032% data of all. The implemented hardware used about 200K gates in $0.25{\mu}m$ CMOS library and was stably operated with 165MHz clock frequency in timing simulation.

[ $PLC-{\gamma}1$ ] for Differentiating Adenocarcinoma from Reactive Mesothelial Cells in Effusions (체강 삼출액의 진단에 있어서 $PLC-{\gamma}1$ 면역 염색의 유용성)

  • Woo, Yeong-Ju;Kim, Sung-Sook
    • The Korean Journal of Cytopathology
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    • v.8 no.2
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    • pp.115-119
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    • 1997
  • Cytologic diagnosis of reactive or malignant effusion is sometimes difficult. Especially, differentiation of benign reactive mesothelial cells from malignant cells in body effusion is more difficult. Recently, immunohistochemistry has been used to diagnose difficult cases. Phospholipase $C(PLC)-{\gamma}1$ is one of the isoenzyme of the PLC which plays central role in signal transduction involving cellular growth, differentiation and transformation by phosphorylating many protein component. Increased expression of $PLC-{\gamma}1$ in human breast carcinoma, colorectal carcinoma and stomach cancers are reported. To evaluate the efficacy of positive $PLC-{\gamma}1$ immunostaining in the diagnosis of malignancy in effusions, paraffin-embedded cell blocks of pleural fluid and ascites from 10 patients(5 metastatic adenocarcinomas, and 5 reactive mesothelial cells) were immunostained with a monoclonal antibody to $PLC-{\gamma}1$. $PLC-{\gamma}1$ immuostained all the adenocarcinomas in cell block(5/5) with intense membrane pattern, however, none of the reactive mesothelial proliferations stained with the diagnostic membrane pattern. Thus, our study strongly supports the conclusion that $PLC-{\gamma}1$ immunopositivity is likely to become a useful adjunct for the diagnosis of malignancy in effusions.

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