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A novel method for high-frequency genome editing in rice, using the CRISPR/Cas9 system (벼에서 CRISPR/Cas9 활용 고빈도 유전자 편집 방법)

  • Jung, Yu Jin;Bae, Sangsu;Lee, Geung-Joo;Seo, Pil Joon;Cho, Yong-Gu;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.44 no.1
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    • pp.89-96
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    • 2017
  • The CRISPR/Cas9 is a core technology that can result in a paradigm for breeding new varieties. This study describes in detail the sgRNA design, vector construction, and the development of a transgenic plant and its molecular analysis, and demonstrates how gene editing technology through the CRISPR/Cas9 system can be applied easily and accurately. CRISPR/Cas9 facilitates targeted gene editing through RNA-guided DNA cleavage, followed by cellular DNA repair mechanisms that introduce sequence changes at the site of cleavage. It also allows the generation of heritable-targeted gene mutations and corrections. Here, we present detailed procedures involved in the CRISPR/Cas9 system to acquire faster, easier and more cost-efficient gene edited transgenic rice. The protocol described here establishes the strategies and steps for the selection of targets, design of sgRNA, vector construction, and analysis of the transgenic lines. The same principles can be used to customize the versatile CRISPR/Cas9 system, for application to other plant species.

Implementation of the Digital Current Control System for an Induction Motor Using FPGA (FPGA를 이용한 유도 전동기의 디지털 전류 제어 시스템 구현)

  • Yang, Oh
    • Journal of the Korean Institute of Telematics and Electronics C
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    • v.35C no.11
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    • pp.21-30
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    • 1998
  • In this paper, a digital current control system using a FPGA(Field Programmable Gate Array) was implemented, and the system was applied to an induction motor widely used as an industrial driving machine. The FPGA designed by VHDL(VHSIC Hardware Description Language) consists of a PWM(Pulse Width Modulation) generation block, a PWM protection block, a speed measuring block, a watch dog timer block, an interrupt control block, a decoder logic block, a wait control block and digital input and output blocks respectively. Dedicated clock inputs on the FPGA were used for high-speed execution, and an up-down counter and a latch block were designed in parallel, in order that the triangle wave could be operated at 40 MHz clock. When triangle wave is compared with many registers respectively, gate delay occurs from excessive fan-outs. To reduce the delay, two triangle wave registers were implemented in parallel. Amplitude and frequency of the triangle wave, and dead time of PWM could be changed by software. This FPGA was synthesized by pASIC 2SpDE and Synplify-Lite synthesis tool of Quick Logic company. The final simulation for worst cases was successfully performed under a Verilog HDL simulation environment. And the FPGA programmed for an 84 pin PLCC package was applied to digital current control system for 3-phase induction motor. The digital current control system of the 3 phase induction motor was configured using the DSP(TMS320C31-40 MHz), FPGA, A/D converter and Hall CT etc., and experimental results showed the effectiveness of the digital current control system.

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Physicochemical Characteristics of Cheonnyuncho Fruit (Opuntia humifusa) Fermented by Leuconostoc mesenteroides SM (Leuconostoc mesenteroides SM을 이용한 천년초(Opuntia humifusa) 열매 발효물의 물리·화학적 특성)

  • Park, Min-Ju;Lee, Sam-Pin
    • Korean Journal of Food Science and Technology
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    • v.45 no.4
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    • pp.434-440
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    • 2013
  • Functional properties of Cheonnyuncho fruit paste (CFP) were enhanced by fermentation, using Leuconostoc mesenteroides SM. The stability of CFP pigment was maintained and microorganisms in crude CFP were sterilized by heating at $80^{\circ}C$ for 30 min. The production of dextran in CFP was increased by increasing the fermentation time and sugar content, resulting in an increase of consistency. The CFP fermented with 20% sucrose contained 4.8% mucilage after fermentation for 2 days. In particular, the production of mucilage was greatly enhanced by fortification with 3% sodium citrate in CFP, resulting in mucilage production of 7%. In addition, the viscous modulus of fermented CFP showed a greater increase than that of the elastic modulus. Thus, it was concluded that CFP fermented by L. mesenteroides SM can be fortified with functional ingredients, such as mucilage and probiotics, with enhanced rheological properties. Therefore fermented CFP can be utilized as the functional ingredients for functional foods.

Design Optimization of Duplex Burnable Poison Rods and Feasibility Evaluation for Core Design (이중구조 가연성독봉 설계안의 최적화 및 노심 핵설계 타당성 평가)

  • Yoon Seok-Kyun;Lee Dae-Jin;Kim Myung-Hyun
    • Journal of Energy Engineering
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    • v.13 no.4
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    • pp.242-258
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    • 2004
  • The duplex burnable poison absorbers concept was suggested by Korea Atomic Energy Research Institute. This BP rod is composed of inner region of natural U-Gd$_2$O$_3$ and outer shell of enriched UO$_2$-Er$_2$O$_3$. It is expected that this burnable absorber has same reactivity control capability with gadolinia burnable absorber used in extened fuel cycle. In order to evaluate the nuclear feasibility of duplex BPs, the nuclear design characteristics were compared with that of four types of burnable absorbers; gadolinia, erbia, IFBA, dysprosia duplex BP on 24 months fuel cycle for Korean Standard Nuclear Power plants. According to the evaluation results of nuclear characteristics, the duplex BPs were better than other BPs on k-infinitives, reactivity holddown worth (RHW), pin power peaking and moderator temperature coefficient (MTC). The possibility of nuclear core design was also confirmed based on the optimized fuel assemblies which were searched for a sensitivity analysis. Characteristics of core design with duplex BPs was compared with that of reference core with gadolinia BPs for cycle length, power peaking and MTC. The duplex BP core had a little longer cycle length by 4 to 7 days because of increased amount of fissile in enriched uranium at the outer shell of duplex BP In case of power peaking F$\_$Q/ of duplex BP core was reduced from 1.5773 to 1.5335. MTC was also less -0.48 pcm/C than that of reference core. Finally, evaluation of fuel cycle economy was performed for the manufacturing feasibility test and fuel cost evaluation with duplex BPs. Fuel cycle economy of duplex BP core almost was equivalent with that of gadolinia BP core.

Long-term performance of drainage system for leakage treatment of tunnel operating in cold region (한랭지역에서 운영 중인 터널의 누수처리를 위한 유도배수시스템의 장기 성능 평가)

  • Kim, Dong-Gyou
    • Journal of Korean Tunnelling and Underground Space Association
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    • v.20 no.6
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    • pp.1177-1192
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    • 2018
  • The objective of this study is to develop the existing drainage system for catching the partial leakage of tunnel structures operating in cold region. The drainage system consists of drainage board, Hotty-gel as a waterproofing material, cover for preventing protrusion of Hotty-gel, air nailer, fixed nail, pipe for collecting ground leak, pipe for conveying ground leak, wire-mesh, and sprayed cement mortar. The drainage systems were installed in conventional concrete lining tunnels to evaluate the site applicability and constructability. The performances of waterproof and the drainage in the drainage system were evaluated by injecting 1,000 ml of red water in the back of the drainage system at 7 days, 14 days, 21 days, 28 days, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months and 8 months. During 8 months of field test, the average daily temperature of the tunnel site was measured from $-16.0^{\circ}C$ to $25.6^{\circ}C$. The daily minimum temperature was $-21.3^{\circ}C$ and the daily maximum temperature was $30.8^{\circ}C$. There was no problem in waterproof and drainage performance of the drainage board in the drainage system. However, the pipe for conveying ground leak had the leakage problem from 14 days. It is considered that the leakage of the pipe for conveying ground leak was caused by the deformation of the pipe of the flexible plastic material having a thickness of 0.2 cm by using the high pressure air nailer and the fixing pin and the insufficient thickness and width of the hotty-gel for preventing the leakage.

Observation of Response to PPD Skin Test and Local Side Reactions at Multiple Inoculation Sites after Percutanous Inoculation with BCG Tokyo 172 Strain (경피용 건조 B.C.G. 백신(Tokyo 172주) 접종 후 국소 반응과 투베르쿨린 양전율에 관한 연구)

  • Lee, Jin Soo;Sohn, Young Mo
    • Pediatric Infection and Vaccine
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    • v.7 no.2
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    • pp.201-210
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    • 2000
  • Purpose : We observed response to PPD skin test and local side reactions among subjects who received inoculation with Tokyo 172 BCG strain by percutaneous method using multiple puncture device. Methods : 138 infants and young children were enrolled at Yongdong Severance Hospital and 7 private clinics. 5TU PPD skin test were performed at 4 months after inoculation. The local reactions at multiple puncture site were observed in 3 days, 4~6 weeks, 36 weeks, and 48 weeks after inoculations and physical check up was done for evaluation of lymphadenopathy. Results : During 48 weeks of observation period, 96 subjects among 138 who were enrolled were followed up completely with records of PPD skin test and observation of local side reactions, presenting with the photos. The size of the induration after 48 hours of PPD skin test, was less than 5mm in six subjects(6.3%), greater than 10mm in sixty seven subjects(70.0%) and greater than 12mm in forty six subjects(47.9%). All subjects showed inflammatory reaction and pustules at multiple puncture sites and only just small papules, ulcer and pustules remained 4-6 weeks later. Eight to twelve weeks later, all local inflammatory skin reactions disappeared with remaining crust. After 48 weeks, 4(4.2%) subjects showed no scar with only faint stain on the puncture site. More than 70% of subjects showed more than 10 faint pin-point scars on the sites. However, the size of scar was clearly smaller compared to that of intradermal inoculation. There were no cases of lymphadenopathy. Conclusion : We observed good immune response to 5TU PPD skin test among the infant and young children who were immunized with percutanous inoculation of Tokyo 172 BCG strain. We could not find any severe local scar at inoculation sites. A degree of satisfaction of the parents whose children received the percutaneous injection was very high.

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Analysis of Complications after Treatment of Calcaneal Fracture (종골 골절 일차 치료 후 발생한 합병증에 대한 분석)

  • Suh, Dong-Hyun;Park, Yong-Wook;Kim, Do-Young;Lee, Sang-Soo;Yoon, Tae-Kyung;Park, Hyun-Chul;Kang, Seung-Wan
    • Journal of Korean Foot and Ankle Society
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    • v.8 no.1
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    • pp.46-51
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    • 2004
  • Purpose: Problematic late sequelae are common following a calcaneal fracture regardless of the initial treatment. We retrospectively evaluated the painful conditions and reviewed the results of the operative treatment in patients with previously treated calcaneal fractures. Materials and Methods: Between October 1996 and September 2001, forty-three patients who underwent subsequent surgical treatment for late sequelae of calcaneal fracture were reviewed. The initial treatment consisted of only immobilization in a cast in 7 patients, closed reduction with pin fixation (Essex-Lopresti technique) in 22 and open reduction and internal fixation in 14. Painful conditions in the hind foot included subtalar arthritis in 31 patients, calcaneofibular impingement in 13, peroneal tendinitis in 6, displaced posterior bony fragment in 3, sural neuritis in 2, subtalar and midtarsal arthritis in 1 and displaced plantar bony fragment in 1. The surgical procedures for the late complications were performed at a mean of 19 months (range, 6 to 35 months) after the injury and consisted of lateral wall ostectomy and in situ subtalar fusion in 28 patients, only lateral wall ostectomy in 5 patients, lateral wall ostectomy and subtalar distraction arthrodesis in 3, removal of displaced posterior bony fragment in 3, sural nerve transposition in the peroneus brevis in 2, triple arthrodesis in 1 and removal of displaced plantar bony fragment in 1. Mean postoperative follow up period was 57 months (range, 33 to 82 months). The results of treatment were evaluated on the basis of pain, improvement in the ability to perform activities of daily living, to return to work or to a pre-injury level of activity. Results: Pain was partially relieved in 38 patients (88%), but not relieved in 5. Function improved in 34 patients (79%), and 32 (74%) returned to work or to a pre-injury level of activity. There was a trend that the longer the interval between the injury and the operation, the longer the subsequent interval until the patient returned to full activities or work. Conclusion: Meticulous physical examination and intensive prompt treatment for remaining pain after initial treatment of calcaneal fractures are recommended for patient's satisfaction and returning to work.

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X-RAY ASTRONOMY EXPERIMENT ON THE INDIAN SATELLITE IRS-P3

  • AGRAWAL P. C.;PAUL B.;RAO A. R.;SHAH M. R.;MCKERJEE K.;VARIA M. N.;YADAV J. S.;DEDHIA D. K.;MALKAR J. P.;SHAH P.;DAMLE S. V.;MARAR T. M. K.;SEETHA S.
    • Journal of The Korean Astronomical Society
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    • v.29 no.spc1
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    • pp.429-432
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    • 1996
  • An x-ray astronomy experiment consisting of three collimated proportional counters and an X-ray Sky Monitor (XSM) was flown aboard the Indian Satellite IRS-P3 launched on March 21, 1996 from SHAR range in India. The Satellite is in a circular orbit of 830 km altitude with an orbital inclination of $98^{\circ}$ and has three axis stabilized pointing capability. Each pointed-mode Proportional Counter (PPC) is a multilayer, multianode unit filled with P-10 gas ($90\%$ Ar + $10\%\;CH_4$) at 800 torr and having an aluminized mylar window of 25 micron thickness. The three PPCs are identical and have a field of view of $2^{\circ}{\times}2^{\circ}$ defined by silver coated aluminium honeycomb collimators. The total effective area of the three PPCs is about 1200 $cm^2$. The PPCs are sensitive in 2-20 keV band. The XSM consists of a pin-hole of 1 $cm^2$ area placed 16 cm above the anode plane of a 32 cm$\times$32 cm position sensitive proportional counter sensitive in 3-8 keV interval. The position of the x-ray events is determined by charge division technique using nichrome wires as anodes. The principal objective of this experiment is to carry out timing studies of x-ray pulsars, x-ray binaries and other rapidly varying x-ray sources. The XSM will be used to detect transient x-ray sources and monitor intensity of bright x-ray binaries. Observations of black-hole binary Cyg X-1 and few other binary sources were carried out in early May and July-August 1996 period. Details of the x-ray detector characteristics are presented and preliminary results from the observations are discussed.

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Development of natural fermented seasoning with Flammulina velutipes powder fortified with γ-aminobutyric acid (GABA) by lactic acid fermentation (팽이버섯(Flammulina velutipes) 분말의 젖산발효를 통한 고농도 γ-aminobutyric acid 함유 천연 발효조미료 개발)

  • Park, Eun-Jin;Lee, Syng-Ook;Lee, Sam-Pin
    • Food Science and Preservation
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    • v.24 no.2
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    • pp.237-245
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    • 2017
  • Lactic acid fermentation of Flammulina velutipes (FV) powder was optimized to produce higher content of ${\gamma}-aminobutyric$ acid (GABA). FV powder (10%) was fermented with 0.5% yeast extract, 1% glucose, 5% mono sodium-L-glutamate (MSG) by Lactobacillus plantarum EJ2014 for 5 days at $30^{\circ}C$. The pH decreased from 6.1 to 4.4 for first 2 days after then increased to 6.2 for following 5 days. While the acidity increased from 0.5% to 1.3% for 2 days, after then decreased to 0.4% for 5 days. Viable cell count showed higher value of $2.2{\times}10^9CFU/mL$ after fermentation for 5 days. In particular, 3.54% MSG as a substrate was completely utilized during lactic acid fermentation, indicating higher 2.31% GABA content. The fermented FV powder showed higher antioxidant properties than that of un-fermented FV power. $IC_{50}$ values of DPPH radical scavenging and ABTS radical scavenging activities were 1.11 mg/mL and 2.58 mg/mL, respectively. Conclusively, natural fermented seasoning from the lactic acid fermentation of 30g of FV powder and 1 g of roasted wheat bran could provide the functional ingredients with 17% GABA, probiotics and dietary fiber, which is used for health food and functional seasoning.

Revision Anterior Cruciate Ligament Surgery Using Hamstring Autograft (슬괵건을 이용한 전방 십자 인대 재 재건술)

  • In Yong;Bahk Won-Jong;Kwon Oh-Soo;Suh Young-Wan;Im Dong-Sun
    • Journal of the Korean Arthroscopy Society
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    • v.7 no.2
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    • pp.183-188
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    • 2003
  • Purpose : The purpose of this study is to evaluate the results of revision surgery for failed anterior cruciate ligament (ACL) reconstruction using quadruple hamstring tendon autograft. Materials and Methods : From May 2000 to July 2002, six patients received ACL revision surgery using quadruple hamstring autograft for failed ACL reconstruction. Femoral tunnels were made 40 mm in depth and fixed with a cross pin and a bioabsorbable interference screw to fill the bone defect. In tibial tunnels, the grafts were fixed with Intrafix(Mitek, Norwood, MA). In case of tibial tunnel widening, additional screw-washer fixation was done. Follow up was at least 12 months postoperatively. Clinical assessments consisted of Lysholm knee scores, International Knee Documentation Committee(IKDC) evaluation form and manual maximal side to side difference using KT-2000 arthrometer. Results : The average Lysholm knee score improved from 77.2 preoperatively to 87.7 postoperatively. At the final IKDC evaluation, 1 case was graded as normal, 4 nearly normal, 1 abnormal. Mean side to side difference of manual maximum anterior displacement using the KT-2000 arthrometer was 1.8mm. The success rate was $83\%$. Conclusion : ACL revision surgery using quadruple hamstring autograft with double fixation is considered good procedure with successful results.

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