• Title/Summary/Keyword: AC System

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High-Level Expression of T4 Endonuclease V in Insect Cells as Biologically Active Form

  • Kang, Chang-Soo;Son, Seung-Yeol;Bang, In-Seok
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1583-1590
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    • 2006
  • T4 endonuclease V (T4 endo V) [EC 3. 1. 25. 1], found in bacteriophage T4, is responsible for excision repair of damaged DNA. The enzyme possesses two activities: a cyclobutane pyrimidine dimer DNA glycosylase (CPD glycosylase) and an apyrimidic/apurinic endonuclease (AP lyase). T4 denV (414 bp cDNA) encoding T4 en do V (138 amino acid) was synthesized and expressed using either an expression vector, pTriEx-4, in E. coli or a baculovirus AcNPV vector, pBacPAK8, in insect cells. The recombinant His-Tag/T4 endo V (rHis-Tag/T4 endo V) protein expressed from bacteria was purified using one-step affinity chromatography with a HiTrap Chelating HP column and used to make rabbit anti-His-Tag/T4 endo V polyclonal antibody for detection of recombinant T4 endo V (rT4 endo V) expressed in insect cells. In the meantime, the recombinant baculovirus was obtained by cotransfection of BacPAK6 viral DNA and pBP/T4 endo V in Spodoptera frugiperda (Sf21) insect cells, and used to infect Sf21 cells to overexpress T4 endo V protein. The level of rT4 endo V protein expressed in Sf21 cells was optimized by varying the virus titers and time course of infection. The optimal expression condition was set as follows; infection of the cells at a MOI of 10 and harvest at 96 h post-infection. Under these conditions, we estimated the amount of rT4 endo V produced in the baculovirus expression vector system to be 125 mg/l. The rT4 endo V was purified to homogeneity by a rapid procedure, consisting of ion-exchange, affinity, and reversed phase chromatographies, based on FPLC. The rT4 endo V positively reacted to an antiserum made against rHis-Tag/T4 endo V and showed a residual nicking activity against CPD-containing DNA caused by UV. This is the first report to have T4 endo V expressed in an insect system to exclude the toxic effect of a bacterial expression system, retaining enzymatic activity.

Effects of Donor Somatic Cell Conditions on In Vitro Development of Nuclear Transplanted Porcine Embryos (돼지 공여세포의 조건이 핵이식 수정란의 체외발달에 미치는 영향)

  • 홍승표;박준규;이명열;이지삼;정장용
    • Journal of Embryo Transfer
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    • v.16 no.3
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    • pp.213-221
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    • 2001
  • This study was conducted to examine in vitro developmental ability of porcine embryos after somatic cell nuclear transfer. The porcine ear fell was cultured in vitro for confluency in serum-starvation condition(TCM-199 + 0.5% FBS) far 3~6 days of cell confluency. The zona pellucida of IVM oocytes were partially drilled using laser system. Single somatic cell was individually transferred into enucleated oocytes. And the reconstructed embryos were electrically fused(single DC 1.9kv/cm, 30$\mu$ sec) with 0.3M mannitol. After electrofusion, embryos were activated(single AC 5v/mm, 5sec) and cultured in HCSU-23 medium containing 10% FBS at 39$^{\circ}C$, 5% $CO_2$ in air for 6 to 8 days. The fusion rate of donor cells was 45.6, 36.8 and 46.1% in 3~4, 5~6 days of serum starvation and non serum starvation(N-S), and were 52.7. 53.0 and 51.7% in 1~2. 5~6 and 13~14 passages of donor cell culture, respectively. No significant difference was found in the fusion rate of donor cells by the duration of serum starvation treatment or the number of donor cell passages. By the size of donor cells, however, the fusion rate was significantly higher(P<0.05) for reconstructed embryos derived from 25r $\mu$m $\geq$ site of donor cells (65.3%) than that of 25~30$\mu$ m(42.5%) or 30$\mu$ m(45.5%)$\leq$ cells. The cleavage rate was significantly (P<0.05) higher in 3~4 darts of serum starvation treatment(67.1%) than that in N-S (50.7%) or 5~6 days of starvation(57.1%). The activation rate by the size of donor cells in fused oocytes was 56.5, 68.8 and 58.5%, respectively, and was not significant.

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Prediction of Potential Risk Posed by a Military Gunnery Range after Flood Control Reservoir Construction (홍수조절지 건설 후 사격장 주변지역의 위해성예측 사례연구)

  • Ryu, Hye-Rim;Han, Joon-Kyoung;Nam, Kyoung-Phile;Bae, Bum-Han
    • Journal of Soil and Groundwater Environment
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    • v.12 no.1
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    • pp.87-96
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    • 2007
  • Risk assessment was carried out in order to improve the remediation and management strategy on a contaminated gunnery site, where a flood control reservoir is under construction nearby. Six chemicals, including explosive chemicals and heavy metals, which were suspected to possess risk to humans by leaching events from the site were the target pollutants for the assessment. A site-specific conceptual site model was constructed based on effective, reasonable exposure pathways to avoid any overestimation of the risk. Also, conservative default values were adapted to prevent underestimation of the risk when site-specific values were not available. The risks of the six contaminants were calculated by API's Decision Support System for Exposure and Risk Assessment with several assumptions. In the crater-formed-area(Ac), the non-carcinogenic risks(i.e., HI values) of TNT(Tri-Nitro-Toluene) and Cd were slightly larger than 1, and for RDX(Royal Demolition Explosives), over 50. The total non-carcinogenic risk of the whole gunnery range calculated to a significantly high value of 62.5. Carcinogenicity of Cd was estimated to be about $10^{-3}$, while that of Pb was about $5\;{\times}\;10^{-4}$, which greatly exceeded the generally acceptable carcinogenic risk level of $10^{-4}{\sim}10^{-6}$. The risk assessment results suggest that an immediate remediation practice for both carcinogens and non-carcinogens are required before the reservoir construction. However, for more accurate risk assessment, more specific estimations on condition shifts due to the construction of the reservoir are required, and more over, the effects of the pollutants to the ecosystem is also necessary to be evaluated.

The Characteristics Analysis and Design of High-Frequency Isolated Type ZVZCS PS-PWM DC-DC Converter with Fuel Cell Generation System (연료전지 발전시스템에 적용된 고주파 절연형 ZVZCS PS-PWM DC-DC 컨버터의 설계 및 특성 해석)

  • Suh, Ki-Young;Mun, Sang-Pil;Kim, Dong-Hun;Lee, Hyun-Woo;Kwon, Soon-Kurl
    • Journal of the Korean Institute of Illuminating and Electrical Installation Engineers
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    • v.20 no.4
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    • pp.21-28
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    • 2006
  • In this paper, the proposed full-bridge high frequency isolated zoo voltage and zero current switching phase shifted pulse width modulation(ZVZCS PS-PWM)DC-DC converter among fuel cell generation system consist of 1.2[kW] fuel cell of Nexa Power Module, full-bridge DC-DC converter to boost the fuel cell low voltage($28{\sim}43[%]$) to 380[VDC] and a single phase full-bridge inverter is implemented to produce AC output(220[VAC], 60[Hz]). A tapped inductor filter with freewheeling diode is newly implemented in the output filter of the proposed full-bridge high frequency isolated ZVZCS PS-PWM DC-DC converter to suppress circulating current under the wide output voltage regulation range, thus to eliminate the switching and transformer turn-on/off over-short voltage or transient phenomena. Besides the efficiency of $93{\sim}97[%]$ is obtained over the wide output voltage regulation ranges and load variations.

Use of Peristeum as a Source of Endothelial-like Cells (혈관내피유사세포 채취의 원천으로 골막의 활용)

  • Park, Bong-Wook;Kim, Shin-Won;Kim, Uk-Kyu;Hah, Young-Sool;Kim, Jin-Hyun;Kim, Deok-Ryong;Sung, Iel-Young;Cho, Yeong-Cheol;Son, Jang-Ho;Kim, Jong-Ryoul;Byun, June-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.33 no.5
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    • pp.385-391
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    • 2011
  • Purpose: The periosteum is a well-known source of osteogenic precursor cells for tissue-engineered bone formation. However, cultured endothelial or endothelial-like cells derived from periosteum have not yet been investigated. This study focused on endothelial-like cell culture from the periosteum. Methods: Periosteal tissues were harvested from the mandible during surgical extraction of lower impacted third molars. The tissues were treated with 0.075% type I collagenase in phosphate-buffered saline (PBS) for 1 hr at $37^{\circ}C$ to release cellular fractions. The collagenase was inactivated with an equal volume of DMEM/10% fetal bovine serum (FBS) and the infranatant was centrifuged for 10 min at 2,400 rpm. The cellular pellet was filtered through a $100{\mu}m$ nylon cell strainer, and the filtered cells were centrifuged for 10 min at 2,400 rpm. The resuspended cells were plated into T25 flasks and cultured in endothelial cell basal medium (EBM)-2. Results: Among the hematopoietic markers, CD146 was more highly expressed than CD31 and CD34. The periosteal-derived cells also expressed CD90 and CD166, mesenchymal stem cell markers. Considering that the expression of CD146 was constant and that the expression of CD90 was lower at passage 5, respectively, the CD146 positive cells in passage 5 were isolated using the magnetic cell sorting (MACS) system. These CD146 sorted, periosteal-derived cells formed tube-like structures on Matrigel. The uptake of acetylated, low-density lipoprotein, labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI-Ac-LDL) was also examined in these cells. Conclusion: These results suggest that the CD146-sorted positive cells can be referred to as periosteal-derived CD146 positive endothelial-like cells. In particular, when a co-culture system with endothelial and osteoblastic cells in a three-dimensional scaffold is used, the use of periosteum as a single cell source would be strongly beneficial for bone tissue engineering.

Treatment of Distal Clavicular Fracture by Reconstruction Plate and Wire Fixation (쇄골 원위부 골절에 대한 재건 금속판과 강선 고정술을 이용한 치료)

  • Suh, Mu-Sam;Kim, Sang-Hyo;Park, Hyeong-Taek
    • Journal of Korean Orthopaedic Sports Medicine
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    • v.5 no.1
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    • pp.58-62
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    • 2006
  • Purpose: To evaluate the result of operative treatment with plate and wire fixation for fractures of the distal clavicle. Materials and Methods: We assessed 9 cases of fractures of the distal clavicle that were treated by plate and wire fixation from March 2001 to July 2003 and followed up over 1year. We used the reconstruction plate as a buttress and performed cerclage wiring to fix comminuted fracture site. The functional result of shoulder was evaluated by the scoring system of Rowe. Results: The average periods of bony union was 9 weeks in all cases. According to Rowe scoring system, the functional result was exellent in 7 cases and good in 2 cases. There was no LOM of shoulder and traumatic arthritis in AC joint. But there was skin irritation in 1 case. Conclusion : The operative method by reconstruction plate and wire fixation for distal clavicular fracture was useful when there were comminuted fracture. This method seems to be simple and satisfactory in terms of improvement of range of motion of shoulder, restoration of function and rare complication.

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Analysis of Environmental Effects for Linear Type Traits and Scoring Traits on Holstein Cows (Holstein 젖소의 선형심사형질과 등급형질에 대한 환경효과 분석)

  • 이득환;김은길
    • Journal of Animal Science and Technology
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    • v.48 no.6
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    • pp.767-776
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    • 2006
  • Corresponding author:Deukhwan Lee, Department of Animal Life Resources, Hankyong National Univ. Seokjeong-dong Anseong-si Gyeonggi-do, Rep. of Korea 456-749Tel: 031-670-5091, Fax: 031-676-5091, E-mail : dhlee@hknu.ac.krGeneral performance with including environmental and management effects on linear type traits in Holstein cows were investigated. 115,646 valid records measured from cows over 1 yrs of age by Korean Animal Improvement Association from 2000 to 2004 were used for this study. Farm, appraisal year-month, appraisal person should affect linear type and scoring traits. Most of type traits and scoring traits would be significantly affected by parity and lactation stage after absorbing farm-appraisal year-month-person effects. Otherwise, some traits such as traits related to udder would be affected by registration criteria. However, interval of appraisal time and milking time would not affect these traits. The scores related to udder, teat placement and foot angle would be positively related to parity. Final score would be optimized at cows of 2nd and 3rd parities. Dairy form, front teat placement, rear udder height, rear udder width and final score would show similar to pattern of lactation curve at lactation stage. Dairy capacity composite index would also show similar to lactation curve. This result would be indicated that more concise standardizing system for linear type scoring rules would be needed. Furthermore, correcting system for parity and lactation stage would be needed because this factor should affect physiological status, specially, udder status.

IP Voltage Controller of Three-phase PWM Converter for Power Supply of Communication System (IP 제어기를 이용한 통신 전원용 3상 PWM 컨버터의 전압 제어)

  • Shin, Hee-Keun;Kim, Hag-Wone;Cho, Kwan-Yuhl;Ji, Jun-Keun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.12 no.6
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    • pp.2722-2728
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    • 2011
  • 3Phase PWM rectifier has become increasingly popular due to its capability of nearly sinusoidal waveform of the input current, and nearly unity power factor operation as a AC/DC rectifier of high capacity telecommunication power supply system. Generally, PI controller is used as a voltage controller of PWM rectifier and voltage controller must be designed to have low overshoot in transient state to get a reliability and stable operation. However, in the application of telecommunication in which load condition is varied very fast, the voltage controller must have a large bandwidth to reduce output voltage variation. The PI controller with large bandwidth arouse the excessive overshoot of the output voltage, and this large output voltage variation degrades the reliability of communication power of the three-phase PWM Rectifier. In this paper, new IP voltage controller for 3 phase PWM rectifier is proposed which has relatively low transient output voltage variation. The improved output characteristics of the transient state voltage responses of the starting and at load changes of the proposed voltage controller are proved by simulations and experiments.

The study on DC-link Film Capacitor in 3 Phase Inverter System for the Consideration of Frequency Response (3상 인버터 시스템에서 주파수 특성을 고려한 필름 콘덴서의 DC-link 적용 방법에 관한 연구)

  • Park, Hyun-Soo
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.4
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    • pp.117-122
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    • 2018
  • A large-capacity three-phase system air conditioner recently includes an inverter circuit to reduce power consumption. The inverter circuit uses a DC voltage that comes from DC-link power capacitor with the function of rectifying, which means AC voltage to DC voltage using a diode. An electrolytic capacitor is generally used to satisfy the voltage ripple and current ripple conditions of a DC-link power capacitor used for rectifying. Reducing the capacitance of the capacitor decreases the size, weight, and cost of the circuit. This paper proposes an algorithm to reduce the input ripple current by combining the minimum point estimation phase locked loop (PLL) phase control and the average voltage d axis current control technique. When this algorithm was used, the input ripple current decreased by almost 90%. The current ripple of the DC-link capacitor decreased due to the decrease in input ripple current. The capacitor capacity can be reduced but the electrolytic capacitor has a heat generation problem and life-time limitations because of its large equivalent series resistance (ESR). This paper proposes a method to select a film capacitor considering the current ripple at DC-link stage instead of an electrolytic capacitor. The capacitance was selected considering the voltage limitation, RMS (Root Mean Square) current capacity, and RMS current frequency analysis. A $1680{\mu}F$ electrolytic capacitor can be reduced to a $20{\mu}F$ film capacitor, which has the benefit of size, weight and cost. These results were verified by motor operation.

Noise Characteristics of 64-channel 2nd-order DROS Gradiometer System inside a Poorly Magnetically-shielded Room (저성능 자기차폐실에서 64채널 DROS 2차 미분계 시스템의 잡음 특성)

  • Kim, J.M.;Lee, Y.H.;Yu, K.K.;Kim, K.;Kwon, H.;Park, Y.K.;Sasada, Ichiro
    • Progress in Superconductivity
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    • v.8 no.1
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    • pp.33-39
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    • 2006
  • We have developed a second-order double relaxation oscillation SQUID(DROS) gradiometer with a baseline of 35 mm, and constructed a poorly magnetically-shielded room(MSR) with an aluminum layer and permalloy layers for magnetocardiography(MCG). The 2nd-order DROS gradiometer has a noise level of 20 $fT/{\surd}Hz$ at 1 Hz and 8 $fT/{\surd}Hz$ at 200 Hz inside the heavily-shielded MSR with a shielding factor of $10^3$ at 1 Hz and $10^4-10^5$ at 100 Hz. The poorly-shielded MSR, built of a 12-mm-thick aluminum layer and 4-6 permalloy layers of 0.35 mm thickness, is 2.4mx2.4mx2.4m in size, and has a shielding factor of 40 at 1 Hz, $10^4$ at 100 Hz. Our 64-channel second-order gradiometer MCG system consists of 64 2nd-order DROS gradiometers, flux-locked loop electronics, and analog signal processors. With the 2nd-order DROS gradiometers and flux-locked loop electronics installed inside the poorly-shielded MSR, and with the analog signal processor installed outside it, the noise level was measured to be 20 $fT/{\surd}Hz$ at 1 Hz and 8 $fT/{\surd}Hz$ at 200 Hz on the average even though the MSR door is open. This result leads to a low noise level, low enough to obtain a human MCG at the same level as that measured in the heavily-shielded MSR. However, filters or active shielding is needed fur clear MCG when there is large low-frequency noise from heavy air conditioning or large ac power consumption near the poorly-shielded MSR.

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