• Title/Summary/Keyword: 0.9 V to 5 V

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Molecular Cloning and Characterization of Mannitol-1-Phosphate Dehydrogenase from Vibrio cholerae

  • Rambhatla, Prashanthi;Kumar, Sanath;Floyd, Jared T.;Varela, Manuel F.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.9
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    • pp.914-920
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    • 2011
  • Vibrio cholerae utilizes mannitol through an operon of the phosphoenolpyruvate-dependent phosphotransferase (PTS) type. A gene, mtlD, encoding mannitol-1-phosphate dehydrogenase was identified within the 3.9 kb mannitol operon of V. cholerae. The mtlD gene was cloned from V. cholerae O395, and the recombinant enzyme was functionally expressed in E. coli as a $6{\times}$His-tagged protein and purified to homogeneity. The recombinant protein is a monomer with a molecular mass of 42.35 kDa. The purified recombinant MtlD reduced fructose 6-phosphate (F6P) using NADH as a cofactor with a $K_m$ of $1.54{\pm}0.1$ mM and $V_{max}$ of $320.8{\pm}7.81\;{\mu}mol$/min/mg protein. The pH and temperature optima for F6P reduction were determined to be 7.5 and $37^{\circ}C$, respectively. Using quantitative real-time PCR analysis, mtlD was found to be constitutively expressed in V. cholerae, but the expression was up-regulated when grown in the presence of mannitol. The MtlD expression levels were not significantly different between V. cholerae O1 and non-O1 strains.

The Crystallinity and Electrical Properties of SrBi2Ta2O9 Thin Films Fabricated by New Low Temperature Annealing (새로운 저온 열처리 공정으로 제조된 SrBi2Ta2O9 박막의 결정성 및 전기적 특성)

  • Lee, Kwan;Choi, Hoon-Sang;Jang, Yu-Min;Choi, In-Hoon
    • Korean Journal of Materials Research
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    • v.12 no.5
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    • pp.382-386
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    • 2002
  • We studied growth and characterization of $SrBi_2Ta_2O_9$ (SBT) thin films fabricated by low temperature process under vacuum and/or oxygen ambient. A metal organic decomposition (MOD) method based on a spin-on technique and annealing process using a rapid thermal annealing (RTA) method was used to prepare the SBT films. The crystallinity of a ferroelectric phase of SBT thin films is related to the oxygen partial pressure during RTA process. Under an oxygen partial pressure higher than 30 Torr, the crystallization temperature inducing the ferroelectric SBT phase can be lowered to $650^{\circ}C$. Those films annealed at $650^{\circ}C$ in vacuum and oxygen ambient showed good ferroelectric properties, that is, the memory window of 0.5~0.9 V at applied voltage of 3~7 V and the leakage current density of 1.80{\times}10^{-8}$ A/$\textrm{cm}^2$ at an applied voltage of 5V. In comparison with the SBT thin films prepared at 80$0^{\circ}C$ in $O_2$ ambient by furnace annealing process, the SBT thin films prepared at $650^{\circ}C$ in vacuum and oxygen ambient using the RTA process showed a good crystallization and electrical properties which would be able to apply to the virtul device fabrication precess.

Determination of saikosaponin derivatives in Bupleuri Radix using HPLC-ELSD (HPLC-ELSD를 이용한 시호 중의 saikosaponin 유도체의 확인법 개발)

  • Kim, Bo-Mi;Yoon, Kee-Dong;Han, Kyung-Reem;Kim, Jin-Woong
    • Journal of Pharmaceutical Investigation
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    • v.38 no.1
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    • pp.57-61
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    • 2008
  • A HPLC-ELSD method was developed to determine saikosaponin derivatives from Bupleuri Radix. Eight saikosaponins, saikosaponin c, i, h, a, $b_2$, g, $b_1$ and d, were analyzed under optimized HPLC conditions [column: Eclipse XDB $C_{18}$ ($150{\times}4.6mm$ i.d., $5{\mu}m$; mobile phase: $H_2O$ with 0.1% $CH_3$COOH (v/v) for solvent A and AcCN with 0.1% $CH_3$COOH (v/v) for solvent B, gradient elution; flow rate: 1mL/min; injection volume: $20{\mu}L$]. Good linearity was achieved in the range from 62.5 to $250{\mu}g/mL$ for each compound, and intra-day precision and accuracy at each concentration level varied between 0.05 and 5.45% and between 93.9 and 109.6%, respectively, whereas those for inter-day variations were between 0.91 to 2.73% and 94.3 to 106.1%. This HPLC-ELSD method was applied for the determination of sakosaponins from Bupleuri Radix samples, and saikosaponin a $(0.79{\pm}0.20mg/g)$, c $(0.33{\pm}0.06mg/g)$ and d $(0.48{\pm}0.15mg/g)$ were observed as major compounds. The other saikosaponins were shown under limit of quantification level thus couldn't be quantified. The present study suggested that the introduced HPLC-ELSD method is selective and reliable, and not only saikosaponin a, but also saikosaponin c and d should be employed as the standard markers for Bupleuri Radix.

Pathogenicity and PCR detection of Vibrio tapetis in Manila clams, Ruditapes philippinarum (양식 바지락, Ruditapes philippinarum에 대한 Vibrio tapetis의 병원성과 PCR법에 의한 진단)

  • Park, Sung-Woo;Lee, Kyung-Hee
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.39-48
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    • 2005
  • Pathogenicity of Vibrio tapetis, the causative bacterium of 'brown ring disease (BRD)' was evaluated in Manila clams (Ruditapes philippinarumi by artificially 0.1 $m\ell$ infection of $1.0\times10^5$cells and $1.0\times10^8$ cells at 20 $^{\circ}C$. A PCR assay based on 16S rRNA to detect the bacteria in clam tissues was established. Accumulative mortality of clams infected with $1.0\times10^7$cells and $1.0\times10^4$ cells per an individual of the bacteria was 67.5% and 7.5%, respectively. However, the deposit of brown pigment in the inner shells by accumulation of chonchiolin was not found. The bacteria were not be able to re-isolate from the infected clams by the conventional agar plate method but were easily detected by PCR assay established in this experiment. In clams artificially infected with 10 species of Vibrio, a 414bp for V. tapetis was detected in PCR assay. The specific band in the clams infected with $1.0\times10^4$cells per an individual of V. tapetis was detected only in gills one day after the infection but never be found in any tissues including gills three days after the infection. In the case of clams infected with $1.0\times10^8$cells per an individual of V. tapetis the specific band was detected in gills and intestine one day after the infection, in all tissues three days after the infection, and then in gills and adductor muscle nine days after the infection. The PCR assay was applied to detect V. tapetis in manila clam, surf clam (Mactra veneriformis), oyster (Crassostrea gigas) and Thomas' rapa whelk (Rapana venosa) taken from Taean and Gochang from April to July 2004. The infection rates were detected to 23.1% and 9.4% in the oyster and surf clam, while manila clam and Thomas' rapa whelk were not found.

Study on the Variations of Inter-annual Tracheid Length for Korean Red Pine from Sokwang-ri in Uljin (경상북도 울진군 소광리 소나무의 가도관 길이 변이 분석)

  • Seo, Jeong-Wook;Eom, Chang-Deuk;Park, So-Yeon
    • Journal of the Korean Wood Science and Technology
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    • v.42 no.6
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    • pp.646-652
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    • 2014
  • This study was carried out to test the possibility that tracheid length of red pine growing in Sokwang-ri in Uljin could be used as one of variables to estimate its wood properties. For the study, forest stands of V (500 m a.s.l.) and VIIII (900 m a.s.l.) age class plots were selected in the region, and three trees in each plot were investigated. The tracheid length was separately measured for early- and latewood. It took approximately 25 and 40 years for V and VIIII age class stands, respectively, until the length was stabilized. The lengths in latewood were 3.14 (V age class) and 3.30 (VIIII age class) mm, and in earlywood 2.98 (V age class) and 3.15 (VIIII age class) mm. The lengths in latewood therefore were longer than in earlywood and the lengths for VIIII age class were longer than for V age class. However, the ratio between their lengths in early- and latewood was the same as 0.96 in all age classes. It might be verified whether this ratio can be used as a variable to test the wood property according to provinces by comparing it with others.

The Comparison of Electric Characteristics of Radiation Detective Sensor(a-Se) with changing composition ratio of Arsenic (Arsenic의 첨가량에 따른 방사선 검출센서 (a-Se)의 전기적 특성 비교)

  • Seok, Dae-Woo;Kang, Sang-Sik;Lee, Dong-Gil;Kim, Jae-Hyung;Nam, Sang-Hee
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2002.11a
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    • pp.391-394
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    • 2002
  • There has recently been much interest and research in developing digital x-ray systems based on using amorphous selenium(a-Se) photoconductors as the image receptor. The amorphous selenium layer that is currently being studied for use as an x-ray photoconductor is not pure a-Se but rather amorphous selenium alloyed with arsenic. We fabricated samples using the selenium and arsenic alloy with various concentrations of the arsenic. In this work, x-ray photoconductor using amorphous selenium alloyed with arsenic were fabricated with different concentrations of the arsenic (0.1 wt.%, 0.3wt.%, 0.5wt.%, 1wt.%, 1.5wt.%, 3wt.%, 5wt.%). The seven kind of samples was fabricated with a-Se alloyed with arsenic through vacuum thermal evaporation. We also investigate the arsenic concentration dependence on the device performance in radiation detector. The electric characteristics of radiation detector devices with changing additive ratio of the arsenic is performed by measuring the x-ray induced photocurrent and integrating it over time to find the total charge. The thickness of a-Se is $100{\mu}m$. Bias voltages $3V/{\mu}m$, $6V/{\mu}m$$9V/{\mu}m$ are applied at the samples. As results, the net charge of a-Se 0.3% As sample is $526.0pC/mR/cm^2$ at $9V/{\mu}m$ bias. The net charge is decreased as with the increasing additive ratio of arsenic.

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Field Application of Biocovers in Landfills for Methane Mitigation (매립지 메탄 저감을 위한 바이오커버의 현장 적용 평가)

  • Jung, Hyekyeng;Yun, Jeonghee;Oh, Kyung Cheol;Jeon, Jun-Min;Ryu, Hee-Wook;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.45 no.4
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    • pp.322-329
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    • 2017
  • Two pilot-scale biocovers (PBCs) were installed in a landfill, and the methane ($CH_4$) concentrations at their inlets and outlets were monitored for 240 days to evaluate the methane removability. Consequently, the packing materials were sampled from the PBCs, and their potential $CH_4$ oxidizing abilities were evaluated in serum vials. The $CH_4$ concentration at the inlet of the biocovers was observed to be in the range of 23.7-47.9% (average = 41.3%, median = 42.6%). In PBC1, where a mixture of soil, earthworm cast, and compost (7:2:1, v/v) was employed as the packing material, the $CH_4$ removal efficiency was evaluated to be between 60.7-85.5%. In PBC2, which was filled with a mixture of soil, earthworm cast, perlite, and compost (4:2:3:1, v/v), the removal efficiency was evaluated to be between 29.2-78.5%. Although the packing materials had an excellent $CH_4$ oxidizing potential (average oxidation rate for $CH_4=180-199{\mu}g\;CH_4{\cdot}g\;packing\;material^{-1}{\cdot}h^{-1}$), $CH_4$ removal efficiency in PBC1 and PBC2 decreased to the range of 0-30% once the packing materials in the PBCs were clogged and channeled. Furthermore, seasonal effects exhibited no significant differences in the $CH_4$ removal efficiency of the biocovers. The results of this study can be used to design and operate real-scale biocovers in landfills to mitigate $CH_4$ buildup.

The Wormicidal Sibstance of Fresh Water Fishes on Clonorchis sinensis(VI) - Struture Identification of Purified Clonorchicidal Sudstance from Epidermal Mucus of Cyprinus carpio nudus (간흡충에 대한 살충성물질에 관한 연구 (VI) - 향어 체표면 점액내 살충성물질의 구조 확인)

  • 이재구;김평길;안병준
    • YAKHAK HOEJI
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    • v.31 no.1
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    • pp.33-39
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    • 1987
  • In order to elucidate the defence mechanism of Cyprinus carpio nudus to Clonorchis sinensis, clonorchicidal substance in the epidermal mucus of this fish was isolated by silica gel column and thin layer chromatography and analyzed for its chemical nature by UV, IR and NMR-spectroscopy. The epidermal mucus of the fish was extracted by ethyl ether and separated into 2 fractions by column chromatography using petroleum ether/chloroform(30/70,v/v) as first solvent and methanol as second solvent. The second fraction with greyish white colour showed clonorchicidal effect. The second was again fractionated into greyish white precipitate and clonorchicidal greenish yellow supernatant fraction, by adding petroleum ether and standing the mixture for 5 days at $5^{\circ}C$. The latter was divided into 7 fractions in column chromatography with acetone/ benzene (10/90, v/v) as carrier. The fraction on equivalent to the spot of Rf. 0.225 value among them disclosed the strongest clonorchicidal effect. By this purification procedure, clonorchicidal substance was purified 154-fold with 0.9% yield from the mucus of the fish, and 10mg of the final fraction killed tne metaceicariae in 22 min. Infra red, nuclear magnetic resonance and ultraviolet spectrometric analysis of the purified substance revealed that the substance is linoleic acid. According to the results of the present studies it seemed that this fish could not be proper intermediate host of Clonorchis sixensis, and that defence mechanism of this fish to the worm seems to be correlated with clonorchicidal substance in epidermal mucus.

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Electroporation and Viability Monitoring Chip for Lung Cancer Cells in Single Channel with Multiple Electric Field Zones (다수의 전기장 분포가 생성되는 단일 미세유로를 이용한 폐암세포 전기천공 및 활성도 분석칩)

  • Kim, Min-Ji;Kim, Tae-Yoon;Cho, Young-Ho
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.36 no.9
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    • pp.901-905
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    • 2012
  • We present an electroporation and viability monitoring chip for lung cancer cells in a single channel with multiple electric field zones. Previous electroporation chips utilized multiple microchannels or electrodes to form multiple electric fields, thus resulting in complex structures. However, the present chip can generate multiple electric fields in a single stepwise microchannel between a pair of electrodes, thus achieving the analysis of both cell electroporation and viability with a simple structure. We demonstrate that the electric field of 0.4 kV/cm results in a maximum percentage of $51.4{\pm}3.0%$ and $26.6{\pm}0.7%$ of viable and electroporated human lung cancer cells, H23 and A549, respectively. The present chip has potential for use in integrated cell chips for transfection studies.

Studies on production of nuclear transplanted mouse embryos (핵이식 마우스 생산에 관한 연구)

  • Lee, Byeong-chun;Jo, Choong-ho;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.151-169
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    • 1993
  • The present study was carried out to investigate the best condition for nuclear-cytoplasm fusion and in vitro culture of nuclear transplanted embryos and to investigate the production of nuclear transplanted offsprings. The nuclei from 2-, 4- and 8-cell mouse embryos were transferred into enucleated 2-cell embryos, and the reconstituted embryos were submitted to direct current(DC) pulses at output voltage of 1.0, 1.5 and 2.0 kV/cm for 100, 150 and $200{\mu}$ sec to induce cell fusion. 1. The culture of intact or zona cut 2-cell embryos in the medium supplemented with cytochalasin B($5{\mu}g/m{\ell}$) and colcemide($0.1{\mu}g/m{\ell}$)for 30 and 60 minutes did not affect the development to later stage. 2. The in vitro developmental rates of group A(a nucleus from one of the blastomeres was removed) and B(electrofusion of group A) were significantly lower than that of control group(p<0.01). 3. When nuclear transplanted embryos were submitted to electrofusion, the significantly higher fusion rates of 2-cell donor nuclei were achieved at the electric field strength of DC 1.5kV/cm for 100 and $150{\mu}$ sec, DC 2.0 kV/cm for $100{\sim}200{\mu}$ sec than DC 1.0 kV/cm for 100 and $150{\mu}$ sec(p<0.01). The significantly higher fusion rates of 4-cell donor nuclei were achieved at DC 2.0 kV/cm for 100 and $150{\mu}$ sec than DC 1.0kV/cm for $100{\sim}200{\mu}$ sec(p<0.01). These fusion rates in 8-cell donor nuclei were 88.7~99.3%. 4. The developmental potency to blastocyst in 2- and 4-cell donor nuclei was significantly higher in DC 1.0 and 2.0 kV/cm for $100{\sim}200{\mu}$ sec treated group and DC 2.0 kV/cm for 150 and $200{\mu}$ sec treated group (p<0.01). The developmental potency to blastocyst in 8-cell donor nuclei was significantly higher in DC 2.0 kV/cm for $100{\mu}$ sec treated group than in DC 1.0 kV/cm for $100{\mu}$ sec treated group and DC 2.0 kV/cm for 150 and $200{\mu}$ sec treated group(p<001). 5. The developmental potency to blastocyst after nuclear transplantation was significantly higher in 2-cell donor nuclei than in 8-cell donor nuclei(p<0.01). 6. The success rate of nuclear injection into enucleated 2-cell embryos was significantly higher in 2-cell donor nuclei than in 4- or 8-cell donor nuclei(p<0.01). 7. The culture time taken for the nuclear transplanted 2-cell embryos to blastocyst stage was significantly longer in 2-cell donor nuclei than in 8-cell donor nuclei(p<0.01). 8. There was no significant difference in the developmental potency of nuclear transplanted embryos within the concentration of EGF at 0 to 15 ng per $m{\ell}$ of BMOC-3 solution. 9. The production rates of offspring after transfer of nuclear transplanted embryos to recipient mouse were significantly higher in 2-cell donor nuclei than in 8-cell donor nuclei(p<0.01).

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