• Title/Summary/Keyword: 2-cell block

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The cytokinesis-block micronucleus assay as a biological dosimeter in irradiated lymphocyte : Comparison of the response of mouse and human (임파구의 미세핵 분석법을 이용한 생물학적 방사선 피폭선량 측정 : 마우스와 인체에서의 반응비교)

  • Kim, Sung-ho;Cho, Chul-koo;Kim, Tae-hwan;Yoo, Seong-yul;Koh, Kyoung-hwan;Yun, Hyong-geun;Koh, Joo-hwan;Choi, Soo-yong
    • Korean Journal of Veterinary Research
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    • v.33 no.3
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    • pp.487-492
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    • 1993
  • 정상인 말초혈액임파구 및 C57BL/6마우스 비장임파구에 $^{60}Co{\gamma}-rays$를 in vitro상태에서 조사한 후 500개 또는 1000개의 cytokinesis-blocked(CB) lymphocytes의 미세핵(micronuclei)의 발생빈도를 측정하였다. 방사선조사량에 따라 미세핵의 발생빈도는 증가하였으며 linear-quadratic model로 측정한 결과 선량반응곡선의 식은 인체의 경우 $Y=(0.31{\pm}0.049)D+(0.0022{\pm}0.0002)D^2+13.19$($r^2=1.000$)이었으며, 마우스의 경우 $Y=(1.31{\pm}0.264)D+(0.0015{\pm}0.0006)+8.7$($r^2=0.988$)이었다(Y는 1000개의 CB cell 당 미세핵발생빈도, D는 cGy로 표시되는 조사선량). 인체 말초혈액임파구에 대한 마우스 비장임파구의 상대적 생물학적 효과(relative biological effectiveness)는 미세핵의 발생율이 세포당 0.05~0.8의 범위에서 $1.84{\pm}0.48$이었다. 미세핵분석법은 인체 및 동물의 방사선 피폭시 간편하고 빠른 생물학적 선량측정법으로 사용될 수 있을 것이다.

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Involvement of Intracellular Ca2+-and PI3K-Dependent ERK Activation in TCDD-Induced Inhibition of Cell Proliferation in SK-N-SH Human Neuronal Cells

  • Yang, Seun-Ah;Lee, Yong-Soo;Jin, Da-Qing;Jung, Jae-Wook;Park, Byung-Chul;Lee, Yoon-Seok;Paek, Seung-Hwan;Jeong, Tae-Cheon;Choi, Han-Gon;Yong, Chul-Soon;Yoo, Bong-Kyu;Kim, Jung-Ae
    • Biomolecules & Therapeutics
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    • v.13 no.2
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    • pp.78-83
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    • 2005
  • 2,3,7,8-Tetrachlorodibenzo-p-dioxin(TCDD) has previously shown to induce neurotoxicity through intracellular $Ca^{2+}$ increase in rat neurons. In this study we investigated the role and signaling pathway of intracellular $Ca^{2+}$ in TCDD-induced inhibition of neuronal cell proliferation in SK-N-SH human neuronal cells. We found that TCDD(10nM) rapidly increased the level of intracellular $Ca^{2+}$, which was completely blocked by the extracellular $Ca^{2+}$ chelation with EGTA (1 mM) or by pretreatment of the cells with the non-selective cation channel blocker. flufenamic acid (200 ${\mu}M$). However, pretreatment of the cells with dantrolene (25 ${\mu}M$) and TMB-8(10 ${\mu}M$), intracellular $Ca^{2+}$-release blockers, or a voltage-sensitive $Ca^{2+}$ channel blocker, varapamil (100 ${\mu}M$), failed to block the TCDD-induced $Ca^{2+}$ increase in the cells. In addition, TCDD induced a rapid and transient activation of phatidvlinositol 3-kinase (PI3K) and extracellular signal-regulated kinase 1/2(ERK1/2), which was ingnificantly blocked by the pretreatment with BAPTA, an intracellular $Ca^{2+}$ chelator, and LY294002, a PI3K inhibitor. Furthermore, inhibitors of PI3K, ERK, or an intracellular $Ca^{2+}$ chelator further potentiated the anti-proliferative effect of TCDD in the cells. Collectively, the results suggest that intracellular $Ca^{2+}$ and PI3K-dependent activation of ERK 1/2 may be involved in the TCDD-induced inhibition of cell proliferation in SK-N-SH human neuronal cells.

($P16^{ink4}$ Methylation in Squamous Cell Carcinoma of the Oral Cavity. (구강 편평세포암종에서 $P16^{ink4}$ 유전자의 Methylation에 대한 연구)

  • Kang, Gin-Won;Kim, Kyung-Wook;Lyu, Jin-Woo;Kim, Chang-Jin
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.22 no.2
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    • pp.164-173
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    • 2000
  • The p16 protein is a cyclin dependent kinase inhibitor that inhibits cell cycle progression from $G_1$ phase to S phase in cell cycle. Many p16 gene mutations have been noted in many cancer-cell lines and in some primary cancers, and alterations of p16 gene function by DNA methylation have been noticed in various kinds of cancer tissues and cell-lines. There have been a large body of literature has accumulated indicating that abnormal patterns of DNA methylation (both hypomethylation and hypermethylation) occur in a wide variety of human neoplasma and that these aberrations of DNA methylation may play an important epigenetic role in the development and progression of neoplasia. DNA methylation is a part of the inheritable epigenetic system that influences expression or silencing of genes necessary for normal differentiation and proliferation. Gene activity may be silenced by methylation of up steream regulatory regions. Reactivation is associated with demethylation. Although evidence or a high incidence of p16 alterations in a variety of cell lines and primary tumors has been reported, that has been contested by other investigators. The precise mechanisms by which abnormal methylation might contribute to carcinogenesis are still not fully elucidated, but conceivably could involve the modulation of oncogene and other important regulatory gene expression, in addition to creating areas of genetic instability, thus predisposing to mutational events causing neoplasia. There have been many variable results of studies of head and neck squamous cell carcinoma(HNSCC). This investigation was studied on 13 primary HNSCC for p16 gene status by protein expression in immunohistochemistry, and DNA genetic/epigenetic analyzed to determine the incidence, the mechanisms, and the potential biological significance of its Inactivation. As methylation detection method of p16 gene, the methylation specific PCR(MSP) is sensitive and specific for methylation of any block of CpG sites in a CpG islands using bisulfite-modified DNA. The genomic DNA is modified by treatment with sodium bisulfate, which converts all unmethylated cytosines to uracil(thymidine). The primers designed for MSP were chosen for regions containing frequent cytosines (to distinguish unmodified from modified DNA), and CpG pairs near the 5' end of the primers (to provide maximal discrimination in the PCR between methylated and unmethylated DNA). The two strands of DNA are no longer complementary after bisulfite treatment, primers can be designed for either modified strand. In this study, 13 paraffin embedded block tissues were used, so the fragment of DNA to be amplified was intentionally small, to allow the assessment of methylation pattern in a limited region and to facilitate the application of this technique to samlples. In this 13 primary HNSCC tissues, there was no methylation of p16 promoter gene (detected by MSP and automatic sequencing). The p16 protein-specific immunohistochemical staining was performed on 13 paraffin embedded primary HNSCC tissue samples. Twelve cases among the 13 showed altered expression of p16 proteins (negative expression). In this study, The author suggested that low expression of p16 protein may play an important role in human HNSCC, and this study suggested that many kinds of genetic mechanisms including DNA methylation may play the role in carcinogenesis.

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Specific Gene Silencing by Single Stranded Large Circular Antisense Molecules

  • Park, Jong-Gu
    • Biomedical Science Letters
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    • v.10 no.2
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    • pp.65-73
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    • 2004
  • I report that single-stranded antisense as a part of large circular (LC-) genomic DNA of recombinant M13 phage exhibits enhanced stability, sequence specific antisense activity, and no need for target site search. A cDNA fragment (708 bp) of rat TNF-$\alpha$ was inserted into a phagemid vector, and TNF-$\alpha$ antisense molecules (TNF$\alpha$-LCAS) were produced as single-stranded circular DNA. When introduced into a rat monocyte/macrophage cell line, WRT7/P2, TNF$\alpha$-LCAS was able to ablate LPS-induced TNF-$\alpha$ mRNA to completion. The antisense effect of TNF$\alpha$-LCAS was shown to be sequence-specific because expressions of three control genes ($\beta$-actin, GAPDH and IL-1$\beta$) were not significantly altered by the antisense treatment. Further, TNF$\alpha$-LCAS was found to be highly efficacious as only 0.1 $\mu$g (0.24 nM) of TNF$\alpha$-LCAS was sufficient to block TNF-$\alpha$ expression in 1$\times10^5$ WRT7/P2 cells. I have also observed specific antisense activity in reduction of NF-$\kappa$B gene expression. The results suggest that an antisense sequence as a part of single-stranded circular genomic DNA has a specific antisense activity.

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Production of (R)-Ethyl-4-Chloro-3-Hydroxybutanoate Using Saccharomyces cerevisiae YOL151W Reductase Immobilized onto Magnetic Microparticles

  • Choo, Jin Woo;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1810-1818
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    • 2015
  • For the synthesis of various pharmaceuticals, chiral alcohols are useful intermediates. Among them, (R)-ethyl-4-chloro-3-hydroxybutanoate ((R)-ECHB) is an important building block for the synthesis of L-carnitine. (R)-ECHB is produced from ethyl-4-chloro-3-oxobutanoate (ECOB) by a reductase-mediated, enantioselective reduction reaction. The Saccharomyces cerevisiae YOL151W reductase that is expressed in Escherichia coli cells exhibited an enantioselective reduction reaction toward ECOB. By virtue of the C-terminal His-tag, the YOL151W reductase was purified from the cell-free extract using Ni2+-NTA column chromatography and immobilized onto Ni2+-magnetic microparticles. The physical properties of the immobilized reductase (Imm-Red) were measured using electron microscopy, a magnetic property measurement system, and a zeta potential system; the average size of the particles was approximately 1 μm and the saturated magnetic value was 31.76 emu/g. A neodymium magnet was used to recover the immobilized enzyme within 2 min. The Imm-Red showed an optimum temperature at 45℃ and an optimum pH at 6.0. In addition, Bacillus megaterium glucose dehydrogenase (GDH) was produced in the E. coli cells and was used in the coupling reaction to regenerate the NADPH cofactor. The reduction/oxidation coupling reaction composed of the Imm-Red and GDH converted 20 mM ECOB exclusively into (R)-ECHB with an e.e.p value of 98%.

The Role of Metal Catalyst on Water Permeation and Stability of BaCe0.8Y0.2O3-δ

  • Al, S.;Zhang, G.
    • Journal of Electrochemical Science and Technology
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    • v.9 no.3
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    • pp.212-219
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    • 2018
  • Perovskite type ceramic membranes which exhibit dual ion conduction (proton and oxygen ion conduction) can permeate water and can aid solving operational problems such as temperature gradient and carbon deposition associated with a working solid oxide fuel cell. From this point of view, it is crucial to reveal water transport mechanism and especially the nature of the surface sites that is necessary for water incorporation and evolution. $BaCe_{0.8}Y_{0.2}O_{3-{\alpha}}$ (BCY20) was used as a model proton and oxygen ion conducting membrane in this work. Four different catalytically modified membrane configurations were used for the investigations and water flux was measured as a function of temperature. In addition, CO was introduced to the permeate side in order to test the stability of membrane against water and $CO/CO_2$ and post operation analysis of used membranes were carried out. The results revealed that water incorporation occurs on any exposed electrolyte surface. However, the magnitude of water permeation changes depending on which membrane surface is catalytically modified. The platinum increases the water flux on the feed side whilst it decreases the flux on the permeate side. Water flux measurements suggest that platinum can block water permeation on the permeate side by reducing the access to the lattice oxygen in the surface layer.

Band gap control by tri-block nanoribbon structure of graphene and h-BN

  • Lee, Ji-U;Jeong, Ga-Un
    • Proceeding of EDISON Challenge
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    • 2015.03a
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    • pp.324-329
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    • 2015
  • First-principles investigations on the hybrid one dementional hexagonal hybrboron-nitride nano ribbons (BNNRs) with a armchair graphene nano-ribbons(AGRNRs), are presented. Electronics properties of the mixed armchair BNC nano-ribbon (BNCNRs) structure show control of a band gap on all cases at the special K-point. And we have studied, the band gap is direct in all cases. The band gap of mixed ABNCNRs could be divided into three groups (${\Delta}3p$, ${\Delta}3p+1$ and ${\Delta}3p+2$) and decrease with the increase of the width. Also these results show similar to the AGNRs case. Different from the band gap value ordering of AGNRs (${\Delta}3p+1$ > ${\Delta}3p$ > ${\Delta}3p+2$), the ordering of ABNCNRs is ${\Delta}3p$ > ${\Delta}3p+1$ > ${\Delta}3p+2$. The discrepancy may come from the differences between the edges of AGRNRs and the boundaries of hybrid BNCNRs. In addition, the bandgap of ABNCNRs are much smaller than those of the corresponding AGNRs. Our results show that the origin of band gap for BNCNRs with armchair shaped edges arises from both quantum confinement effect of the edges. These results similar to thecase of AGNRs. These properties of hybrid BN/C nano-ribbon structure may offer suitable bandgap to develop nnanoscale electronics and solar cell beyond individual GNRs and BNNRs.

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The Crystal and Molecular Structure of $N_1$-Cyclohexyl-$N_1$-(o-Chlorobenzal) Imino Thiourea

  • Koo, Chung-Hoe;Kim, Hojing;Kim, Hoon-Sup;Chang, Chong-Whan
    • YAKHAK HOEJI
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    • v.21 no.3
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    • pp.146-158
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    • 1977
  • N$_{1}$-cyclohexyl-$N_{2}$-(o-chlorobenzal) imino thiourea, $C_{14}$H$_{18}$N$_{3}$SCI, crystallizes in $C_{2}$/c, with a=19.68, b=7.74, c=20.42$\AA$, ${\beta}$=$92.$8^{\circ}$ and eight formula units in the unit cell. The structure was solved by the study of Patterson sections, calculated from three-dimensional film data, and was refined by block-diagonal least-squares methods to R=0.16 based on 1288 independent intensity data. The rest atoms of N$_{1}$-cyclohexyl-$_{2}$-(o-chlorobenzal) imino thiourea molecule excluding cyclohexan ring and chlorine atoms approximately lie on a plane. A pair of molecules related by the symmetry centers are connected directly with the N-H.......S hydrogen bonds. Apart from the hydrogen bonding system the structure is held together by the van der Waals forces.

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BONE REGENERATION WITH ADIPOSE TISSUE-DERIVED MESENCHYMAL STEM CELL AND HA/TCP (HA/TCP 골이식재상에 이식된 지방유래 줄기세포의 골모세포로의 분화 및 골형성에 대한 연구)

  • Rim, Jae-Suk;Gwon, Jong-Jin;Jang, Hyon-Seok;Lee, Eui-Seok;Jeong, You-Min;Lee, Tai-Hyung;Park, Jeong-Kyun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.32 no.2
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    • pp.97-106
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    • 2010
  • Aim of the study: An alternative source of adult stem cells that could be obtained in large quantities, under local anesthesia, with minimal discomfort would be advantageous. Adipose tissue could be processed to obtain a fibroblast-like population of cells or adipose tissue-derived stromal cells (ATSCs). This study was performed to confirm the availability of ATSCs in bone tissue engineering. Materials amp; Methods: In this study, adipose tissue-derived mesenchymal stem cell was extracted from the liposuctioned abdominal fat of 24-old human and cultivated, and the stem cell surface markers of CD 105 and SCF-R were confirmed by immunofluorescent staining. The proliferation of bone marrow mesenchymal stem cell and ATSCs were compared, and evaluated the osteogenic differentiation of ATSCs in a specific osteogenic induction medium. Osteogenic differentiation was assessed by von Kossa and alkaline phosphatase staining. Expression of osteocyte specific BMP-2, ALP, Cbfa-1, Osteopontin and osteocalcin were confirmed by RT-PCR. With differentiation of ATSCs, calcium concentration was assayed, and osteocalcin was evaluated by ELISA (Enzyme-linked immunosorbant assay). The bone formation by 5-week implantation of HA/TCP block loaded with bone marrow mesenchymal stem cells and ATSCs in the subcutaneous pocket of nude mouse was evaluated by histologic analysis. Results: ATSCs incubated in the osteogenic medium were stained positively for von Kossa and alkaline phosphatase staining. Expression of osteocyte specific genes was also detected. ATSCs could be easily identified through fluorescence microscopy, and bone formation in vivo was confirmed by using ATSC-loaded HA/TCP scaffold. Conclusions: The present results show that ATSCs have an ability to differentiate into osteoblasts and formed bone in vitro and in vivo. So ATSCs may be an ideal source for further experiments on stem cell biology and bone tissue engineering.

Design of Cryptographic Processor for Rijndael Algorithm (Rijndael 암호 알고리즘을 구현한 암호 프로세서의 설계)

  • 전신우;정용진;권오준
    • Journal of the Korea Institute of Information Security & Cryptology
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    • v.11 no.6
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    • pp.77-87
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    • 2001
  • This paper describes a design of cryptographic processor that implements the Rijndael cipher algorithm, the Advanced Encryption Standard algorithm. It can execute both encryption and decryption, and supports only 128-bit block and 128-bit keys. As the processor is implemented only one round, it must iterate 11 times to perform an encryption/decryption. We implemented the ByteSub and InvByteSub transformation using the algorithm for minimizing the increase of area which is caused by different encryption and decryption. It could reduce the memory size by half than implementing, with only ROM. We estimate that the cryptographic processor consists of about 15,000 gates, 32K-bit ROM and 1408-bit RAM, and has a throughput of 1.28 Gbps at 110 MHz clock based on Samsung 0.5um CMOS standard cell library. To our knowledge, this offers more reduced memory size compared to previously reported implementations with the same performance.