• Title/Summary/Keyword: $A_2$

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Establishment of the expression system of human HtrA2 in the zebrafish (Zebrafish 동물모델에서 human HtrA2의 expression system 정립에 관한 연구)

  • Cho, Sung-Won;Park, Hyo-Jin;Kim, Goo-Young;Nam, Min-Kyung;Kim, Ho-Young;Ko, In-Ho;Kim, Cheol-Hee;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.4
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    • pp.571-578
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    • 2006
  • HtrA2/Omi, a mitochondrial trypsin-like serine protease, is pivotal in regulating apoptotic cell death. Several lines of recent evidence suggest that HtrA2 is associated with the pathogenesis of neurodegenerative disorders; however, the physiological function of HtrA2 still remains elusive. For studying physiological function of HtrA2 in depth, it is necessary to develop a suitable expression system in the model animal. We therefore utilized the zebrafish as a model animal to establish expression of human HtrA2 (hHtrA2) in vivo. For expression of mature HtrA2 as GFP fusion in zebrafish embryos, the HtrA2 (WT) or (S306A) cDNAs with the C-terminal GFP tag were inserted into the pCS2+ plasmid. Expression patterns of HtrA2 in HEK293 cells were first monitored by immunofluorescence staining and immunoblot assays, showing approximately 64 kDa of the HtrA2-GFP fusion proteins. Subsequently, the hHtrA2 plasmid DNA or in vitro transcribed mRNA was microinjected into zebrafish embryos. The expression patterns of HtrA2 in Zebrafish embryos were monitored by GFP fluorescence in 24 hours-post-fertilization (hpf). Although expression patterns of HtrA2-GFP in developing embryos were different between the injected DNA and mRNA, both nucleic acids revealed good expression levels to further study the physiological role of HtrA2 in vivo. This study provides a suitable condition for expressing hHtrA2 in the zebrafish embryos as well as a method for generating useful system to investigate physiological properties of the specific human genes.

Neuroprotective effects of modified Bo-Yang-Hwan-Oh-Tang in N2a neuroblastoma cells (가감보양환오탕(加減補陽還五湯)의 N2a 뇌신경세포에 대한 보호효과)

  • Lim, Kyu;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.77-84
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    • 2006
  • Objectives : To evaluate the neuroprotective effects of modified Bo-Yang-Hwan-O-Tang (BHT), we investigated the neuronal death protection effects to oxidative damages in N2a neuroblastoma cells. Methods : To study the cytotoxic effect of BHT on N2a neuronal cells, the cell viability was determined by MTT assay. To investigate the neuronal death protection of BHT, N2a neuronal cells were induced oxidative damages by H2O2, and assayed the cell viability and DNA fragmentation. We also investigated DPPH free radical scavenging effects of BHT by tube test. Results : In MTT assay, $500{\mu}g/ml$ of BHT was not showed cytotoxic effect on N2a neuronal cells. BHT protected N2a neuronal cells from H2O2-induced cell death in a dose-dependent manner. BHT also protected N2a neuronal cells from H2O2-induced DNA fragmentation. BHT scavenged DPPH free radicals in a dose-dependent manner. Conclusion : These data suggest that BHT may have strong anti-oxidant effects through the free radical scavenging and neuroprotective effects in neuronal cells.

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Relationship between Poor Immunogenicity of HLA-A2-Restricted Peptide Epitopes and Paucity of Naïve $CD8^+$ T-Cell Precursors in HLA-A2-Transgenic Mice

  • Choi, Yoon Seok;Lee, Dong Ho;Shin, Eui-Cheol
    • IMMUNE NETWORK
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    • v.14 no.4
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    • pp.219-225
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    • 2014
  • We examined the immunogenicity of H-2 class I-restricted and HLA-A2-restricted epitopes through peptide immunization of HLA-A2-transgenic mice that also express mouse H-2 class I molecules. All four of the tested epitopes restricted by H-2 class I robustly elicited T-cell responses, but four of seven epitopes restricted by HLA-A2 did not induce T-cell responses, showing that HLA-A2-restricted peptide epitopes tend to be poorly immunogenic in HLA-A2-transgenic mice. This finding was confirmed in HLA-A2-transgenic mice infected with a recombinant vaccinia virus expressing hepatitis C virus proteins. We examined the precursor frequency of epitope-specific naïve $CD8^+$ T cells in HLA-A2-transgenic and conventional C57BL/6 mice and found that the poor immunogenicity of HLA-A2-restricted peptide epitopes is related to the paucity of naïve $CD8^+$ T-cell precursors in HLA-A2-transgenic mice. These results provide direction for the improvement of mouse models to study epitope repertoires and the immunodominance of human T-cell responses.

Panaxadiol and Panaxatriol from Panax ginseng C.A. Meyer Inhibit the Synthesis of Thromboxane $A_2$ in Adrenaline-Stimulated Human Platelet Aggregations (Panax ginseng C.A. Meyer의 PD와 PT는 아드레날린에 의해 유인된 사람 혈소판의 응집반응에서 Thromboxane $A_2$의 생성을 저해한다)

  • Park, Kyeong-Mee;Rhee, Man-Whee;Park, Hwa-Jin
    • Journal of Ginseng Research
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    • v.18 no.1
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    • pp.44-48
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    • 1994
  • In adrenaline-stimulated human platelets, panaxadiol (PD) and panaxatriol (PT) from Panax ginseng C.A. Meyer did not inhibit the $Ca^{2+}$-innux, but inhibited the formation of thromboxane $A_2$ and the platelet aggregations. It seems that PD and PT block a pathyway interconvefing arachidonic acids (20:4) to thromboxane $A_2$ (TX $A_2$), because the amount of $Ca^{2+}$ which phospholipase C or phospholipase $A_2$ requires to liberate 20 : 4 from membrane phospholipids was increased by PD and PT. These results mean that PH and PT have an antiplatelet effect by Inhibiting the formation of TX $A_2$.

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COMPACT INTERPOLATION ON AX = Y IN A TRIDIAGONAL ALGEBRA ALGL

  • Kang, Joo-Ho
    • Journal of applied mathematics & informatics
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    • v.19 no.1_2
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    • pp.447-452
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    • 2005
  • Given operators X and Y on a Hilbert space H, an interpolating operator is a bounded operator A such that AX = Y. In this article, we investigate compact interpolation problems for vectors in a tridiagonal algebra. Let L be a subspace lattice acting on a separable complex Hilbert space H and Alg L be a tridiagonal algebra. Let X = $(x_{ij})\;and\;Y\;=\;(y_{ij})$ be operators acting on H. Then the following are equivalent: (1) There exists a compact operator A = $(x_{ij})$ in AlgL such that AX = Y. (2) There is a sequence {$\alpha_n$} in $\mathbb{C}$ such that {$\alpha_n$} converges to zero and $$y_1\;_j=\alpha_1x_1\;_j+\alpha_2x_2\;_j\;y_{2k}\;_j=\alpha_{4k-1}x_{2k\;j}\;y_{2k+1\;j}=\alpha_{4k}x_{2k\;j}+\alpha_{4k+1}x_{2k+1\;j}+\alpha_{4k+2}x_{2k+2\;j\;for\;all\;k\;\epsilon\;\mathbb{N}$$.

Synthesis of Quinazoline 4-one Derivatives from 2-Aminobenzamide (III) -Reaction with Acid Anhydrides- (2-Aminobenzamide로부터 Quinazoline 4-one계 유도체의 합성(III) -Acid anhydride와의 반응-)

  • Suh, Myung-Eun
    • YAKHAK HOEJI
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    • v.34 no.2
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    • pp.133-138
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    • 1990
  • The reaction of 2-aminobenzamide with phthalic acid anhydride In dioxane produced a bicyclic product 2,8-dioxoisoindole(1,2,a) quinazoline (I) in addition to hydrolysis product 2(2-Carboxyphenyl)-1,2-2H-quinazoline-4-one (II). The yields were 64% and 30% respectively. On the other hand, the same reaction in DMF afforded compound (I) and 2(2N-dimethyl carbamyl phenyl)-1,4-2H-quinazoline-4-one (III) in 30% and 60% yield respectively. The compound III was also obtained by the reaction of compound II with dimethylamine. However the reaction of 2-aminobenzamide with neat succinic acid anhydride gave only bicyclic product 2,8-oxopyrrolidine (2,1,a)-1,4-2H-quinazoline (IV) in 93%.

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The Crystal Structure of Fluoxymesterone, $C_{20}H_{29}FO_3$ (Fluoxymesterone, $C_{20}H_{29}FO_3$의 결정구조)

  • Kim, Ui-Seong;Park, Gwon-Il;Jo, Seong-Il
    • Korean Journal of Crystallography
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    • v.4 no.1
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    • pp.6-10
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    • 1993
  • 11 β ,17 β -dihydroxy-9a-fluoro-l7a-methyl androst-4-en-3-one (Fluoxymesterone), CgoH29 FO,, orthorhombic, P2,2,2,, a=13.468(5) A, b= 19.554 (2)A, c=6.578(9)A, a=b=r=90˚, A (CuKa)=1.5406 A , Dm=1.289cm-3, Dc=1.299cm-3 and Z=4 at T=298k. The structure was solved by direct method using seminvariants of ggg Parity group and refined by the full-matrix least-square method, resulting model with reliability factor R=0.069 for 1098 unique reflection over 3σ . Ring A is an 1β-2a-half chair, 5 ring has a highly symmetrical chair conformation, C ring is in a distorted chair conformation and D ring is a 13aenveLope conformation. In the crystal structure, the molecules are packed with a hydrogen bond of 011-H23‥‥03(0.5+x, 1.5-y, 1.0-z) [1.94(9) A of H‥‥0.2.786(9)A of 0‥‥0 and 165(8) ˚ of

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Densification Behavior of Reaction-Bonded Silicon Nitride Prepared by Using Coarse Si Powders (조대 Si입자분말을 사용한 질화반응 Si3N4의 치밀화 거동)

  • 이주신;문지훈;한병동;박동수;김해두
    • Journal of the Korean Ceramic Society
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    • v.39 no.1
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    • pp.45-50
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    • 2002
  • Effect of sintering additives on the densification behavior of reaction-bonded silicon nitride prepared by using coarse Si powders is discussed. Sintering additives such as 6 wt% $Y_2O_3$+1wt% $A1_2O_3$ (6YlA) did not give rise to full densification, while full densification was obtained by using the sintering additives such as 6wt% $Y_2O_3$+3 wt% $A1_2O_3$+ 2wt% $SiO_2$ (6Y3A2S) and 9wt% $Y_2O_3$+ 1.5wt% $A1_2O_3$+ 3wt% $SiO_2$ (9Yl.5A3S). In the case of 6Y3A2S addition, high fracture strength of 960 MPa and the fracture toughness of $6.5 MPa.m^{1/2}$ were obtained.

Effects of CYP1A2$^*$1C and CYP1A2$^*$1F Genotypes on the Activity and Inducibility of CYP1A2 Determined by Urinary Caffeine Metabolite Ratio in Koreans

  • Shin, Mi-Kyung;Yi, Hyeon-Gyu;Kwon, Yong-Hyun;Lee, Sung-Keun;Lim, Woo-Sung;Park, Chang-Shin;Kang, Ju-Hee
    • Molecular & Cellular Toxicology
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    • v.3 no.4
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    • pp.314-319
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    • 2007
  • The effects of common variants of CYP1A2 gene (CYP1A2$^*$1C and CYP1A2$^*$1F) on the CYP1A2 activity and inducibility were controversial. The aim of the present study is to investigate the effects of CYP1A2$^*$1C and CYP1A2$^*$1F on the activity of CYP1A2 determined by urinary caffeine metabolite ratio in Koreans. As might be expected, there was large inter-individual variation (16-folds) of CYP1A2 activity ranged from 2.41 to 39.58. The mean CYP1A2 activity of smokers was significantly higher than that of non-smokers. The frequencies of CYP1A2$^*$1C (-3858A) and $^*$1F (-164A) alleles were 0.219 and 0.646, respectively. The effect of CYP1A2$^*$1C on the CYP1A2 activity was not significant. However, the CYP1A2 activity of subjects with AA genotype for CYP1A2$^*$1F allele was significantly lower than that of non-AA genotypes (CC, or CA). Interestingly, the significant effect of CYP1A2$^*$1F allele on CYP1A2 activity was not observed in nonsmokers. Our results suggest that CYP1A2$^*$1F allele rather than CYP1A2$^*$1C allele significantly influences on the inducibility of CYP1A2 in Koreans. Owing to small sample size of our study, further studies should be conducted to reveal the inter-ethnic difference or the gene-environmental interaction.

Expression of Recombinant Human Cytochrome P450 1A2 in Escherichia coli Bacterial Mutagenicity Tester Strain

  • Chun, Young-Jin
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.305-309
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    • 1998
  • Human cytochrome P450 1A2 is one of the major cytochrome P450s in human liver. It is known to be capable of activating a number of carcinogens such as arylamines and heterocyclic amines. In order to develop the new bacterial mutagenicity test system with human P450, a full length of human P450 1A2 cDNA inserted into pCW bacterial expression vector was introduced to Escherichia coli WP2 uvrA strain which is a well-known E. coli strain for bacterial reverse mutagenicity assay. Expressed human P450 1A2 showed typical P450 hemoprotein spectra. Maximum expression was achieved at 48 hrs after incubating at $30^{\circ}C$ in terrific broth containing ampicillin, IPTG and other supplements. High level expression of P450 1A2 in E. coli WP2 uvrA membranes was determined in SDS-PAGE. The well-known mutagens 2-aminoanthracene and MElQ increased the revertant colonies of E. coli WP2 uvrA expressing human P450 1A2 without an exogenous rat hepatic post-mitochondrial supernatant (S9 fraction) in a dose-dependent manner. The results show that the functional expression of human P450 in bacterial mutagenicity tester strain will provide a useful tool for studying the mechanism of the mutagenesis and carcinogenesis of new drugs and environmental chemicals.

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