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Effects of Sperm Membrane Disruption and Electrical Activation of Oocytes on In vitro Development and Transgenesis of Porcine Embryos Produced by Intracytoplasmic Sperm Injection

  • Shim, Sang Woo;Kim, Young Ha;Lee, Hoon Taek;Shim, Hosup
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.3
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    • pp.358-363
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    • 2008
  • The intracytoplasmic sperm injection (ICSI) procedure has recently been utilized to produce transgenic animals and may serve as an alternative to the conventional pronuclear microinjection in species such as pigs whose ooplasm is opaque and pronuclei are often invisible. In this study, the effects of sperm membrane disruption and electrical activation of oocytes on in vitro development and expression of transgene green fluorescent protein (GFP) in ICSI embryos were tested to refine this recently developed procedure. Prior to ICSI, sperm heads were treated with Triton X-100+NaCl or Triton X-100+NaCl+NaOH, to disrupt membrane to be permeable to exogenous DNA, and incubated with linearized pEGFP-N1 vector. To induce activation of oocytes, a single DC pulse of 1.3 kV/cm was applied to oocytes for $30{\mu}sec$. After ICSI was performed with the aid of a micromanipulator, in vitro development of embryos and GFP expression were monitored. The chemical treatment to disrupt sperm membrane did not affect the developmental competence of embryos. 40 to 60% of oocytes were cleaved after injection of sperm heads with disrupted membrane, whereas 48.6% (34/70) were cleaved without chemical treatment. Regardless of electrical stimulation to induce activation, oocytes were cleaved after ICSI, reflecting that, despite sperm membrane disruption, the perinuclear soluble sperm factor known to mediate oocyte activation remained intact. After development to the 4-cell stage, 11.8 (2/17, Triton X-100+NaCl+NaOH) to 58.8% (10/17, Triton X-100+NaCl) of embryos expressed GFP. The expression of GFP beyond the stage of embryonic genome activation (4-cell stage in the pig) indicates that the exogenous DNA might have been integrated into the porcine genome. When sperm heads were co-incubated with exogenous DNA following the treatment of Triton X-100+NaCl, GFP expression was observed in high percentage (58.8%) of embryos, suggesting that transgenic pigs may efficiently be produced using ICSI.

Surface Modification and Heat Treatment of Ti Rod by Electro Discharge (전기방전에 의한 Ti rod의 열처리 및 표면개질 특성에 관한 연구)

  • Byun, C.S.;Oh, N.H.;An, Y.B.;Cheon, Y.W.;Kim, Y.H.;Cho, Y.J.;Lee, C.M.;Lee, W.H.
    • Korean Journal of Materials Research
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    • v.16 no.3
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    • pp.168-172
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    • 2006
  • Single pulse of 2.0 to 3.5 kJ from $150{\mu}F$ capacitor was applied to the cp Ti rod for its surface modification and heat treatment. Under the conditions of using 2.0 and 2.5 kJ of input energy, no phase transformation has been occurred. However, the hardness and tensile strength decreased and the elongation increased after a discharge due to a slight grain growth. By using more than 3.0 kJ of input energy, the electro discharge made a phase transformation and the hardness at the edge of the cross section increased significantly. The Ti rod before a discharge was lightly oxidized and was primarily in the form of $TiO_2$. However, the surface of the Ti rod has been instantaneously modified by a discharge into the main form of TiN from $TiO_2$. Therefore, the electro discharge can modify its surface chemistry in times as short as $200{\mu}sec$ by manipulating the input energy, capacitance, and discharging environment.

Surface Characteristics of Porous Ti-6Al-4V Implants Fabricated by Electro-Discharge-Sintering in a Low Vacuum Atmosphere (저진공 분위기 전기방전소결에 의해 제조된 다공성 Ti-6Al-4V 임플란트의 표면특성 연구)

  • Hyun, C.Y.;Huh, J.K.;Lee, W.H.
    • Korean Journal of Materials Research
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    • v.16 no.3
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    • pp.178-182
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    • 2006
  • A single electro-discharge-sintering (EDS) pulse (1.0 kJ/0.7 g), from a $300{\mu}F$ capacitor, was applied to atomized spherical Ti-6Al-4V powder in a low vacuum to produce porous-surfaced implant compacts. A solid core surrounded by a porous layer was formed by a discharge in the middle of the compact. XPS (X-ray photoelectron spectroscopy) was used to study the surface characteristics of the implant material. C, O, and Ti were the main constituents, with smaller amounts of Al, V, and N. The implant surface was lightly oxidized and was primarily in the form of $TiO_2$ with a small amount of metallic Ti. A lightly etched EDS implant sample showed the surface form of metallic Ti, indicating that EDS breaks down the oxide film of the as-received Ti-6Al-4V powder during the discharge process. The EDS Ti-6Al-4V implant surface also contained small amounts of aluminum oxide in addition to $TiO_2$. However, V detected in the EDS Ti-6Al-4V implant surface, did not contribute to the formation of the oxide film..

An Implementation of Mobile Respiration Detection Diagnostic System Using Ultrasound Sensing Method (초음파 센싱 방식의 이동형 호흡 측정 진단 시스템의 구현)

  • 김동학;김영길;정승호
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2003.05a
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    • pp.514-517
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    • 2003
  • Oxygen supply is one of the most basic things in human body. Breathing is controlled by the lungs' stationary function and the respiratory center which is in the mesulla oblongata. Nothing but, the external breathing that air movement between the lungs and atmosphere and the internal breathing that cellular air movement between the hemoglobin and a single cell. The adult's number of times of the respirations is about 15∼20 per 1 minute, but it depends ages, exercise, temperature, disease, etc. The important thing in detecting the respiration is that doing it in object person's resting time. Detecting the respiration have to be done without attracting any attention of object person. In present using method is detecting the pulse with catching an object person's wrist and observing the object person's movement. In this paper, we propose the mobile respiration detection diagnostic system using ultrasound sensing method that does not be influenced by the inertia error and the pressure error.

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[$Na^+-Ca^{2+}$ Exchange Curtails $Ca^{2+}$before Its Diffusion to Global $Ca^{2+}{_i}$ in the Rat Ventricular Myocyte

  • Ahn, Sung-Wan;Ko, Chang-Mann
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.2
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    • pp.95-101
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    • 2005
  • In the heart, $Na^{+}-Ca^{2+}$ exchange (NCX) is the major $Ca^{2+}$ extrusion mechanism. NCX has been considered as a relaxation mechanism, as it reduces global $[Ca^{2+}]_i$ raised during activation. However, if NCX locates in the close proximity to the ryanodine receptor, then NCX would curtail $Ca^{2+}$ before its diffusion to global $Ca^{2+}_i$ This will result in a global $[Ca^{2+}]_i$ decrease especially during its ascending phase rather than descending phase. Therefore, NCX would decrease the myocardial contractility rather than inducing relaxation in the heart. This possibility was examined in this study by comparing NCX-induced extrusion of $Ca^{2+}$ after its release from SR in the presence and absence of global $Ca^{2+}_i$ transient in the isolated single rat ventricular myocytes by using patch-clamp technique in a whole-cell configuration. Global $Ca^{2+}_i$ transient was controlled by an internal dialysis with different concentrations of BAPTA added in the pipette. During stimulation with a ramp pulse from +100 mV to -100 mV for 200 ms, global $Ca^{2+}_i$ transient was suppressed only mildly, and completely at 1 mmol/L, and 10 mmol/L BAPTA, respectively. In these situations, ryanodine-sensitive inward NCX current was compared using $100{\mu}mol/L$ ryanodine, $Na^+$ depletion, 5 mmol/L $NaCl_2$ and $1{\mu}mol/L$ nifedipine. Surprisingly, the result showed that the ryanodine-sensitive inward NCX current was well preserved after 10 mmol/L BAPTA to 91 % of that obtained after 1 mmol/L BAPTA. From this result, it is concluded that most of the NCX-induced $Ca^{2+}$ extrusion occurs before the $Ca^{2+}$ diffuses to global $Ca^{2+})i$ in the rat ventricular myocyte.

Synthesis of Titanium Silicide by Electro-Discharge-Sintering of Ti and Si Powder Mixture (Ti 및 Si 혼합 분말의 전기방전소결에 의한 Titanium Silicide의 합성 연구)

  • Cheon Y. W.;Oh N. H.;Kim Y. H.;Byun C. S.;Lee S. H.;Lee W. H.
    • Journal of Powder Materials
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    • v.12 no.6 s.53
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    • pp.447-452
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    • 2005
  • The synthesis and consolidation of titanium silicide by electro-discharge-sintering has been investigated. As-received Ti powder was in flaky shape and the mean particle size was $45.0{\mu}m$, whereas the mean particle size of the pre-milled Si powder with angular shape was $8.0{\mu}m$. Single pulse of 2.5 to 5.0 kJ/0.34g-elemental Ti and pre-milled Si powder mixture with the composition of $Ti-37.5at.\%$ Si was applied using $300{\mu}F$ capacitor. The solid with $Ti_5Si_3$ phase has been successfully fabricated by the discharge with the input energy more than 2.5kJ in less than $129{\mu}sec.$ Hv values were found to be higher than $1000kgf/mm^2$. The formation of $Ti_5Si_3$ occurred through a fast solid state diffusion reaction.

A Study of Electro-Discharge-Sintering of Ti-6Al-4V Spherical Powders Doped with Hydroxyapatite by Spex Milling and Its Consolidation Characteristics (Hydroxyapatite가 도핑된 Ti-6Al-4V 구형 분말의 전기방전 소결 및 소결체 특성에 관한 연구)

  • Cho, Y.J.;Kim, Y.H.;Jo, Y.H.;Kim, M.J.;Kim, H.S.;Kim, S.W.;Park, J.H.;Lee, W.H.
    • Journal of Powder Materials
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    • v.20 no.5
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    • pp.376-381
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    • 2013
  • Spherical Ti-6Al-4V powders in the size range of 250 and 300 ${\mu}m$ were uniformly doped with nano-sized hydroxyapatite (HAp) powders by Spex milling process. A single pulse of 0.75-2.0 kJ/0.7 g of the Ti-6Al-4V powders doped with HAp from 300 mF capacitor was applied to produce fully porous and porous-surfaced Ti-6Al-4V implant compact by electro-discharge-sintering (EDS). The solid core was automatically formed in the center of the compact after discharge and porous layer consisted of particles connected in three dimensions by necks. The solid core increased with an increase in input energy. The compressive yield strength was in a range of 41 to 215 MPa and significantly depended on input energy. X-ray photoelectron spectroscopy and energy dispersive x-ray spectrometer were used to investigate the surface characteristics of the Ti-6Al-4V compact. Ti and O were the main constituents, with smaller amount of Ca and P. It was thus concluded that the porous-surfaced Ti-6Al-4V implant compacts doped with HAp can be efficiently produced by manipulating the milling and electro-discharge-sintering processes.

Voltage-Dependent Inactivation of Calcium Currents in the Mouse Eggs

  • Park, Young-Geun;Yang, Young-Seon;Yum, Myung-Kul;Hong, Seong-Geun
    • The Korean Journal of Physiology
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    • v.25 no.2
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    • pp.125-131
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    • 1991
  • Inactivation properties of Ca current in the unfertilized eggs of mouse were studied by using the whole cell voltage clamp technique and single microelectrode voltage clamp technique. Membrane potential was held at -80 mV and step depolarization was applied from -50 mV to 50 mV for $200{\sim}500\;ms$. Peak of inward Ca currents was $-2{\sim}-4\;nA$ at a membrane Potentials from -20 mV to 0 mV and outward currents were not observed within the membrane voltage range studied $(-50{\sim}50\;mV)$. Inward currents were fully inactivated within 200 ms after the onset of step depolarization. As the membrane became depolarized, time constant of inactivation (${\tau}$) was decreased but remained around $20{\sim}30\;ms$ beyond 10 mV. When $Ca^{2+}$ was used as a charge earlier, inactivation of inward $Ca^{2+}$ current also occured and time course of inactivation was similar to that of $Ca^{2+}$ currents as charge carrier. In the bathing solution containing high potassium $(131\;mM\;K^+)$, process of inactivation was not changed except a parallel decrease of value for the entire range of membrane potential. Steady-state inactivation of the $current(h_{\infty})$ obtained from the double pulse experiment showed the voltage-dependent change. These results suggested that inactivation of Ca currents in the unfertilized eggs of mouse was voltage-dependent.

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A study on the PWM(pulse witdh modulation) current source Inverter with utility (태양광발전 연계 시스템에 의한 PWM 전류형 인버어터에 관한 연구)

  • Hwang, Lak-Hoon;Choi, Ho-Kyu;Sin, Yang-Ho;Lee, Chun-Sang;Kim, Ju-Rae;Jo, Sang-Rou;Jo, Moon-Taeck
    • Proceedings of the KIEE Conference
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    • 2001.07b
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    • pp.1020-1022
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    • 2001
  • because the output of solar cell is direct, it is necessary to install D/A converter system for A.C load, and in case of driving utility line system, it is possible to drive system relation when the system supplies sinusodal current ant voltage having unit power factor. As the characteristics of the soar cell output the is influenced by dailysunight charge, for more electric power it is essential to control the direction toward the san so that the driving point of solar cell can always operate near maximum output point. PWM modulation device among electric power converters must have stable modulation at anytime when it includes noise-factors such as noise-wave and noises on electric voltage wave, a synchronous signal system. In dealing with synchronous signal for control and control signal by microprocessor, it is necessary to compensate it because there is time difference between sample paint and carrier wave. On this papers, single phase PWM current type invertor controled the solar cell having typical voltage dropping character has optimun short current in short, reduces D.C reactance, composes controller for modulation and keeps lower harmonic and high power factor keeping maximum output of solar cell according daily sunlight charge variation.

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Ultra-Low Powered CNT Synaptic Transistor Utilizing Double PI:PCBM Dielectric Layers (더블 PI:PCBM 유전체 층 기반의 초 저전력 CNT 시냅틱 트랜지스터)

  • Kim, Yonghun;Cho, Byungjin
    • Korean Journal of Materials Research
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    • v.27 no.11
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    • pp.590-596
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    • 2017
  • We demonstrated a CNT synaptic transistor by integrating 6,6-phenyl-C61 butyric acid methyl ester(PCBM) molecules as charge storage molecules in a polyimide(PI) dielectric layer with carbon nanotubes(CNTs) for the transistor channel. Specifically, we fabricated and compared three different kinds of CNT-based synaptic transistors: a control device with $Al_2O_3/PI$, a single PCBM device with $Al_2O_3/PI:PCBM$(0.1 wt%), and a double PCBM device with $Al_2O_3/PI:PCBM$(0.1 wt%)/PI:PCBM(0.05 wt%). Statistically, essential device parameters such as Off and On currents, On/Off ratio, device yield, and long-term retention stability for the three kinds of transistor devices were extracted and compared. Notably, the double PCBM device exhibited the most excellent memory transistor behavior. Pulse response properties with postsynaptic dynamic current were also evaluated. Among all of the testing devices, double PCBM device consumed such low power for stand-by and its peak current ratio was so large that the postsynaptic current was also reliably and repeatedly generated. Postsynaptic hole currents through the CNT channel can be generated by electrons trapped in the PCBM molecules and last for a relatively short time(~ hundreds of msec). Under one certain testing configuration, the electrons trapped in the PCBM can also be preserved in a nonvolatile manner for a long-term period. Its integrated platform with extremely low stand-by power should pave a promising road toward next-generation neuromorphic systems, which would emulate the brain power of 20 W.