Purpose: $[^{11}C]6-OH-BTA-1$ ([N-methyl-$^{11}C$]2-(4'-methylaminophenyl)-6-hydroxybenzothiazole, 1), a -amyloid imaging agent for the diagnosis of Alzheimer's disease in PET, can be labeled with higher yield by a simple loop method. During the synthesis of $[^{11}C]1$, we found the formation of by-products in various solvents, e.g., methylethylketone (MEK), cyclohexanone (CHO), diethylketone (DEK), and dimethylformamide (DMF). Materials and Methods: In Automated radiosynthesis module, 1 mg of 4-aminophenyl-6-hydroxybenzothiazole (4) in 100 l of each solvent was reacted with $[^{11}C]methyl$ triflate in HPLC loop at room temperature (RT). The reaction mixture was separated by semi-preparative HPLC. Aliquots eluted at 14.4, 16.3 and 17.6 min were collected and analyzed by analytical HPLC and LC/MS spectrometer. Results: The labeling efficiencies of $[^{11}C]1$ were $86.0{\pm}5.5%$, $59.7{\pm}2.4%$, $29.9{\pm}1.8%$, and $7.6{\pm}0.5%$ in MEK, CHO, DEK and DMF, respectively. The LC/MS spectra of three products eluted at 14.4, 16.3 and 17.6 mins showed m/z peaks at 257.3 (M+1), 257.3 (M+1) and 271.3 (M+1), respectively, indicating their structures as 1, 2-(4'-aminophenyl)-6-methoxybenzothiazole (2) and by-product (3), respectively. Ratios of labeling efficiencies for the three products $([^{11}C]1:[^{11}C]2:[^{11}C]3)$ were $86.0{\pm}5.5%:5.0{\pm}3.4%:1.5{\pm}1.3%$ in MEK, $59.7{\pm}2.4%:4.7{\pm}3.2%:1.3{\pm}0.5%$ in CHO, $9.9{\pm}1.8%:2.0{\pm}0.7%:0.3{\pm}0.1%$ in DEK and $7.6{\pm}0.5%:0.0%:0.0%$ in DMF, respectively. Conclusion: The labeling efficiency of $[^{11}C]1$ was the highest when MEK was used as a reaction solvent. As results of mass spectrometry, 1 and 2 were conformed. 3 was presumed.
Chung, Dawoon;Barker, Bridget M.;Carey, Charles C.;Merriman, Brittney;Werner, Ernst R.;Lechner, Beatrix E.;Dhingra, Sourabh;Cheng, Chao;Xu, Wenjie;Blosser, Sara J.;Morohashi, Kengo;Mazurie, Aurelien;Mitchell, Thomas K.;Haas, Hubertus;Mitchell, Aaron P.;Cramer, Robert A.
한국균학회소식:학술대회논문집
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2015.05a
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pp.15-15
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2015
Aspergillus fumigatus is a major cause of invasive aspergillosis (IA), a significant health issue worldwide with high mortality rates up to 95%. Our lab is interested in how A. fumigatus adapts to low oxygen conditions 'hypoxia', which is one of the important host microenvironments. A. fumigatus SrbA is a basic helix-loop-helix (bHLH) transcriptional regulator and belongs to sterol regulatory element binding protein (SREBP) family members. Loss of SrbA completely blocks growth in hypoxia and results in avirulence in murine models of IA suggesting an essential role of SrbA in hypoxia adaptation and virulence in A. fumigatus. We conducted chromatin immunoprecipitation sequencing (ChIP-seq) with A. fumigatus wild type using a SrbA specific antibody, and 97 genes were revealed as SrbA direct targets. One of the 'SrbA regulons' (AFUB_099590) was a putative bHLH transcriptional regulator whose sequence contained a characteristic tyrosine substitution in the basic portion of the bHLH domain of SREBPs. Therefore, we designated AFUB_099590 SrbB. Further characterization of SrbB demonstrated that SrbB is important for radial growth, biomass production, and biosynthesis of heme intermediates in hypoxia and virulence in A. fumigatus. A series of quantitative real time PCR showed that transcription of several SrbA regulons is coordinately regulated by two SREBPs, SrbA and SrbB in hypoxia. This suggests that SrbA and SrbB have both dependent and independent functions in regulation of genes responsible for hypoxia adaptation in A. fumigatus. Together, our data provide new insights into complicated roles of SREBPs in adaptation of host environments and virulence in pathogenic fungi.
Network expansion has been an inevitable method for most traffic equilibrium assignments to consider intersection movements such as intersection delays. The drawback of network expansion is that because it dramatically increases network sizes to emulate possible directional movements as corresponding links, not only is complexities for building network amplified, but computational performance is shrunk. This paper Proposes a new variational inequality formulation for a user-optimal traffic equilibrium assignment model to explicitly consider directional delays without building expanded network structures. In the formulation, directional delay functions are directly embedded into the objective function, thus any modification of networks is not required. By applying a vine-based shortest Path algorithm into the diagonalization algorithm to solve the problem, it is additionally demonstrated that various loop-related movements such as U-Turn, P-Turn, etc., which are frequently witnessed near urban intersections, can also be imitated by blocking some turning movements of intersections. The proposed formulation expects to augment computational performance through reduction of network-building complexities.
Paik Soon-Young;Ra Kyung Soo;Cho Hoon Sik;Koo Kwang Bon;Baik Hyung Suk;Lee Myung Chul;Yun Jong Won;Choi Jang Won
Journal of Microbiology
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v.44
no.1
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pp.64-71
/
2006
To investigate the effects of the nucleotide sequences in Shine-Dalgarno (SD) and the spacer region (SD-ATG) on bovine growth hormone (bGH) gene expression, the expression vectors under the control of the T7 promoter (pT7-7 vector) were constructed using bGH derivatives (bGH1 & bGH14) which have different 5'-coding regions and were induced in E. coli BL21 (DE3). Oligonucleotides containing random SD sequences and a spacer region were chemically synthesized and the distance between the SD region and the initiation codon were fixed to nine bases in length. The oligonucleotides were annealed and fused to the bGH1 and bGH14 cDNA, respectively. When the bGH gene was induced with IPTG in E. coli BL21(DE3), some clones containing only bGH14 cDNA produced considerable levels of bGH in the range of $6.9\%\;to\;8.5\%$ of total cell proteins by SDS-PAGE and Western blot. Otherwise, the bGH was not detected in any clones with bGH1 cDNA. Accordingly, the nucleotide sequences of SD and the spacer region affect on bGH expression indicates that the sequences sufficiently destabilize the mRNA secondary structure of the bGH14 gene. When the free energy was calculated from the transcription initiation site to the +51 nucleotide of bGH cDNA using a program of nucleic acid folding and hybridization prediction, the constructs with values below -26.3 kcal/mole (toward minus direction) were not expressed. The constructs with the original sequence of bGH cDNA also did not show any expression, regardless of the free energy values. Thus, the disruption of the mRNA secondary structure may be a major factor regulating bGH expression in the translation initiation process. Accordingly, the first stem-loop among two secondary structures present in the 5'-end region of the bGH gene should be disrupted for the effective expression of bGH.
Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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2003.07b
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pp.851-856
/
2003
We have investigated the material and electrical properties of $Bi_{4-x}La_xTi_3O_{12}$ (BLT) ferroelectric thin film for ferroelectric nonvolatile memory applications of capacitor type and single transistor type. The 120nm thick BLT films were deposited on $Pt/Ti/SiO_2/Si$ and $SiO_2/Nitride/SiO_2$ (ONO) substrates by the sol-gel spin coating method and were annealed at $700^{\circ}C$. It was observed that the crystallographic orientation of BLT thin films were strongly affected by the excess Bi content and the intermediate rapid thermal annealing (RTA) treatment conditions regardeless of two type substrates. However, the surface microstructure and roughness of BLT films showed dependence of two different type substrates with orientation of (111) plane and amorphous phase. As increase excess Bi content, the crystallographic orientation of the BLT films varied drastically in BLT films and exhibited well-crystallized phase. Also, the conversion of crystallographic orientation at intermediate RTA temperature of above $450^{\circ}C$ started to be observed in BLT thin films with above excess 6.5% Bi content and the rms roughness of films is decreased. We found that the electrical properties of BLT films such as the P-V hysteresis loop and leakage current were effectively modulated by the crystallographic orientations change of thin films.
For the reasonable use of low grade-shallow geothermal energy by Standing Column Well(SCW) system, the basic requirements are depth-wise increase of earth temperature like $2^{\circ}C$ per every 100m depth, sufficient amount of groundwater production being about 10 to 30% of the design flow rate of GSHP with good water quality and moderate temperature, and non-collapsing of borehole wall during reinjection of circulating water into the SCW. A closed loop type-vertical ground heat exchanger(GHEX) with $100{\sim}150m$ deep can supply geothermal energy of 2 to 3 RT but a SCW with $400{\sim}500m$ deep can provide $30{\sim}40RT$ being equivalent to 10 to 15 numbers of GHEX as well requires smaller space. Being considered as an alternative of vertical GHEX, many numbers of SCW have been widely constructed in whole country without any account for site specific hydrogeologic and geothermal characteristics. When those are designed and constructed under the base of insufficient knowledges of hydrgeothermal properties of the relevant specific site as our current situations, a bad reputation will be created and it will hamper a rational utilization of geothermal energy using SCW in the near future. This paper is prepared for providing a guideline of SCW design comportable to our hydrogeothermal system.
Background: To evaluate HPV testing by Hybrid Capture II (HCII) in conjunction with cytology in detecting the residual/recurrence disease after treatment of high-grade cervical intraepithelial neoplasia (CIN II-III) with loop electrosurgical excision procedure (LEEP). Materials and Methods: A retrospective review of 158 patients with histologically confirmed CIN II-III who underwent LEEP between January 2011 and October 2012 was conducted. Post-treatment control was scheduled at the 3rd, 6th, 12th and 18th month. All patients were followed up by Pap smear and HR-HPV genotype and viral load testing. Results: Pre-treatment, HR-HPV DNA, was detected in all specimens of the patients. At follow-up, 25 patients were diagnosed as the residual/recurrent disease during the FU visit, among whom, 16 patients with positive margin: 13 patients (52%) with HR-HPV DNA+/cytology+, 2 patients (8%) with HR-HPV DNA+/cytology-, 1 patient (4%) with cytology+/HR-HPV DNA-; 9 patients with clean margin - 5 patients (55.6%) with HR-HPV DNA+/cytology+; 2 patients (22.2%) with HRHPV DNA+/cytology-, 2 patients (22.2%) with cytology+/HR-HPV DNA-. None of them persisting HR-HPV DNA-/cytology-with positive or negative margin was identified as the residual/recurrent disease. The majority of residual/recurrent disease was detected at the 12th and 18th month FU, and there was almost no difference in the sensitivity and negative predictive value (NPV) between at the 3rd month and the 6th month FU visits. 14 residual/recurrence disease (14/46:30.4%) had pre-treatment high viral load (>5 000 RUL/PC) and 11 (11/112, 9.8%) with pre-treatment low viral load, P<0.05. Conclusions: (1) The persistence HR-HPV DNA is the root cause of the residual/recurrent disease for the women treated for high-grade CIN; the pre-treatment viral load and margin can be seen as the predictor. (2) The FU visit beginning at the 6th month post-treatment and lasting at least 24 months with the combination of cytology and HPV testing. (3) Patients with high pre-treatment HPV load, which is considered as one risk of developing the residual/recurrent disease, should be paid more attention (especially above 500RUL/PC) to by clinicians.
Background: N-myc downstream regulated gene 2 (NDRG2) is a member of the NDRG gene family. Our previous report indicated a possible role for NDRG2 in regulating the cytokine, interleukin-10 (IL-10), which is an important immunosuppressive cytokine. Several pathways, including p38-MAPK, NF-${\kappa}B$, and JAK/STAT, are used for IL-10 production, and the JAK/STAT pathway can be inhibited in a negative feedback loop by the inducible protein, SOCS3. In the present study, we investigated the effect of NDRG2 gene expression on IL-10 signaling pathway that is modulated via SOCS3 and STAT3. Methods: We generated NDRG2-overexpressing U937 cell line (U937-NDRG2) and treated the cells with PMA to investigate the role of NDRG2 in IL-10 production. U937 cells were also transfected with SOCS3- or NDRG2-specific siRNAs to examine whether the knockdown of SOCS3 or NDRG2 influenced IL-10 expression. Lastly, STAT3 and SOCS3 induction was measured to identify the signaling pathway that was associated with IL-10 production. Results: RT-PCR and ELISA assays showed that IL-10 was increased in U937-mock cells upon stimulation with PMA, but IL-10 was inhibited by overexpression NDRG2. After PMA treatment, STAT3 phosphorylation was decreased in a time-dependent manner in U937-mock cells, whereas it was maintained in U937-NDRG2 cells. SOCS3 was markedly reduced in U937-NDRG2 cells compared with U937-mock cells. IL-10 production after PMA stimulation was reduced in U937 cells when SOCS3 was inhibited, but this effect was less severe when NDRG2 was inhibited. Conclusion: NDRG2 expression modulates SOCS3 and STAT3 activity, eventually leading to the inhibition of IL-10 production.
$BiFeO_3/Pb(Zr_{0.52}Ti_{0.48})O_3$(BFO/PZT) multilayer thin films have been prepared on a Pt/Ti/$SiO_2$/Si(100) substrate by chemical solution deposition. BFO single layer, BFO/PZT bilayer and multilayer thin films were studied for comparison. X-ray diffraction analysis showed that the crystal structure of all films was multi-orientated perovskite phase without amorphous and impurity phase. The leakage current density at 500 kV/cm was reduced by approximately four and five orders of magnitude by bilayer and multilayer structure films, compared with BFO single layer film. The low leakage current density leads to saturated P-E hysteresis loops of bilayer and multilayer films. In BFO/PZT multlayer film, saturated remanent polarization of $44.3{\mu}C/cm^2$ was obtained at room temperature at 1 kHz with the coercive field($2E_c$) of 681.4 kV/cm.
Proceedings of the Korean Vacuum Society Conference
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2012.02a
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pp.431-432
/
2012
In the era of 20 nm scaled semiconductor volume manufacturing, Microelectronics Manufacturing Engineering Education is presented in this paper. The purpose of microelectronic engineering education is to educate engineers to work in the semiconductor industry; it is therefore should be considered even before than technology development. Three Microelectronics Manufacturing Engineering related courses are introduced, and how undergraduate students acquired hands-on experience on Microelectronics fabrication and manufacturing. Conventionally employed wire bonding was recognized as not only an additional parasitic source in high-frequency mobile applications due to the increased inductance caused from the wiring loop, but also a huddle for minimizing IC packaging footprint. To alleviate the concerns, chip bumping technologies such as flip chip bumping and pillar bumping have been suggested as promising chip assembly methods to provide high-density interconnects and lower signal propagation delay [1,2]. Aluminum as metal interconnecting material over the decades in integrated circuits (ICs) manufacturing has been rapidly replaced with copper in majority IC products. A single copper metal layer with various test patterns of lines and vias and $400{\mu}m$ by $400{\mu}m$ interconnected pads are formed. Mask M1 allows metal interconnection patterns on 4" wafers with AZ1512 positive tone photoresist, and Cu/TiN/Ti layers are wet etched in two steps. We employed WPR, a thick patternable negative photoresist, manufactured by JSR Corp., which is specifically developed as dielectric material for multi- chip packaging (MCP) and package-on-package (PoP). Spin-coating at 1,000 rpm, i-line UV exposure, and 1 hour curing at $110^{\circ}C$ allows about $25{\mu}m$ thick passivation layer before performing wafer level soldering. Conventional Si3N4 passivation between Cu and WPR layer using plasma CVD can be an optional. To practice the board level flip chip assembly, individual students draw their own fan-outs of 40 rectangle pads using Eagle CAD, a free PCB artwork EDA. Individuals then transfer the test circuitry on a blank CCFL board followed by Cu etching and solder mask processes. Negative dry film resist (DFR), Accimage$^{(R)}$, manufactured by Kolon Industries, Inc., was used for solder resist for ball grid array (BGA). We demonstrated how Microelectronics Manufacturing Engineering education has been performed by presenting brief intermediate by-product from undergraduate and graduate students. Microelectronics Manufacturing Engineering, once again, is to educating engineers to actively work in the area of semiconductor manufacturing. Through one semester senior level hands-on laboratory course, participating students will have clearer understanding on microelectronics manufacturing and realized the importance of manufacturing yield in practice.
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