The purpose of this study is to examined the electrofusion and activation conditions for the production of porcine somatic cell nuclear transfer (SCNT) embryos. In this study, immature oocytes were cultured in TCM-199 with and without hormones for 22 hours. Skin fibroblasts cells of porcine were transferred into the perivitelline space of enucleated in vitro matured oocytes. Cell fusion was performed with two different pulses that each one pulse (DC) of 1.1 kV/cm or 1.5 kV/cm for $30{\mu}sec$. After fusion subsequent activation were divided into three groups; non-treatment (control) and treatment with 2 mM 6-DMAP or $7.5{\mu}g/ml$ cytochalasin B for 4 hours. Transferred embryos were cultured in PZM-3 (Porcine Zygote Medium-3) in $5%\;CO_2$ and 95% air at $39^{\circ}C$ for 7 day. Apoptosis-related genes (Caspase-3, BCL-2, mTOR, and MMP-2) were analyzed by immunofluorescence staining. There was no significant difference between two different electrofusion stimuli in the cleavage rate; $64.9{\pm}4.8%$ in 1.1 kV/cm and $62.7{\pm}4.0%$ in 1.5 kV/cm. However, blastocyst formation rate (%) was significantly different among three different activation groups (no treatment, 2 mM 6-DMAP or $7.5{\mu}g/ml$ cytochalasin B) combined with electrofusion of 1.1 kV/cm. The blastocyst formation rate was $12.6{\pm}2.5$, $20.0{\pm}5.0$, and $34.9{\pm}4.3%$ in control, 2 mM 6-DMAP, and $7.5{\mu}g/ml$ cytochalasin B, respectively. Immunofluorescence data showed that expression levels of caspase-3 in SCNT embryos undeveloped to blastocyst stage were higher than those in the blastocyst stage embryos. Expression levels of Bcl-2 in blastocyst stage embryos were higher than those in the arrested SCNT embryos. These results showed that the combination of an electric pulse (1.1 kV/cm for $30{\mu}sec$) and $7.5{\mu}g/ml$ cytochalasin B treatment was effective for production of the porcine SCNT embryos.
Proceedings of the Korean Society of Medical Physics Conference
/
2002.09a
/
pp.400-403
/
2002
The fundamental experiments for measuring soft x-ray characteristics from the vacuum capillary are described. These experiments were primarily performed in order to generate line spectra such as x-ray lasers. The generator consists of a high-voltage power supply, a polarity-inversion ignitron pulse generator, a turbo-molecular pump, and a radiation tube with a capillary. A high-voltage condenser of 200 nF in the pulse generator is charged up to 20 kV by the power supply, and the electric charges in the condenser are discharged to the capillary in the tube after closing the ignitron. During the discharge, weakly ionized plasma forms on the inner and outer sides of a capillary. In the present work, the pump evacuates air from the tube with a pressure of about 1 mPa, and a demountable capillary was developed in order to measure x-ray spectra according to changes in the capillary length. In this capillary, the anode (target) and cathode elements can be changed corresponding to the objectives. The capillary diameter is 2.0 mm, and the length is adjusted from 1 to 50 mm. When a capillary with aluminum anode and cathode electrodes was employed, both the cathode voltage and the discharge current almost displayed damped oscillations. The peak values of the voltage and current increased when the charging voltage was increased, and their maximum values were -10.8 kV and 4.7 kA, respectively. The x-ray durations observed by a 1.6 ${\mu}$m aluminum filter were less than 30 ${\mu}$s, and we detected the aluminum characteristic x-ray intensity using a 6.8 ${\mu}$m aluminum filter. In the spectrum measurement, two sets of aluminum and titanium electrodes were employed, and we observed multi-line spectra. The line photon energies seldom varied according to changes in the condenser charging voltage and to changes in the electrode element. In the case where the titanium electrode was employed, the line number decreased with corresponding decreases in the capillary length. Compared with incoherent visible light, these rays from the capillary were diffracted and diffused greatly after passing through two slits.
Indium tin oxide (ITO) provides high electrical conductivity and transparency at visible and near-IR wavelengths. ITO is widely used as a transparent electrode for the fabrication of LCDs, OLEDs, and many kinds of optical applications. It is widely employed for electrodes in various electric and display sectors because of its transparency in the visible range and high conductivity. Therefore, one issue is removing a specific area of a layer of material such as ITO or metallic film on a substrate, without affecting the properties of the substrate. ITO-on-glass removal using a laser is friendlier to the environment than traditional methods. In this study, ablation of ITO film on glass using a femtosecond-laser micromachining system (wavelength 1026 nm, pulse duration 150 fs) and a nanosecond-laser micromachining system (wavelength 1027 nm, pulse duration 5 ns) are described, compared, and analyzed.
An, Joonghyun;Youn, Jiae;Cho, Jeonghun;Park, Daejin
Journal of the Institute of Electronics and Information Engineers
/
v.52
no.12
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pp.99-108
/
2015
The embedded microcontroller which is operated by the logic gates synchronized on the clock pulse, is gradually used as main controller of mission-critical systems. Severe electrical situations such as high voltage/frequency surge may cause malfunctioning of the clock source. The tolerant system operation is required against the various external electric noise and means the robust design technique is becoming more important issue in system clock failure problems. In this paper, we propose on-chip backup clock change architecture for the automatic clock failure detection. For the this, we adopt the edge detector, noise canceller logic and glitch-free clock changer circuit. The implemented edge detector unit detects the abnormal low-frequency of the clock source and the delay chain circuit of the clock pulse by the noise canceller can cancel out the glitch clock. The externally invalid clock source by detecting the emergency status will be switched to back-up clock source by glitch-free clock changer circuit. The proposed circuits are evaluated by Verilog simulation and the fabricated IC is validated by using test equipment electrical field radiation noise
Journal of the korean academy of Pediatric Dentistry
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v.29
no.2
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pp.159-167
/
2002
The purpose of this study was to evaluate the efficacy and safety depending on the route of flumazenil, as an antagonist against midazolam. The subjects of this study were 15 volunteers of $22{\sim}24$ years old. They were sedated with midazolam 0.2mg/Kg intranasal spray, and then 40 minutes after midazolam administration, they were given flumazenil 0.2mg intranasal spray for their reversal. For evaluation of the efficacy and safety of intranasal spray for flumazenil, they were monitored with pulse-oxymeter(Nellcor symphony N-3000, Nellcor Puritan CO. USA) and electric sphygmomanometer (Heartcare 200, National CO. Japan), and were assessed themselves using visual analogue scale(VAS) for tranquilization, sleep, fatigue and attitude. All of these subjects were reduced completely without any undesired situations. The results from this study can be summarized as follows ; 1. Nasaly administered flumazenil using spray device produced much more rapid reduction than intravenously administered flumazenil, but soon after fell in more deep sedated state than intravenously administered flumazenil. 2. There were no considerable side effects or bad influence on vital signs of both nasaly administered flumazenil and intravenously administered flumazenil. These results suggested that the flumazenil administered nasaly using spray device for reversal, we could treat patients safely and effectively under conscious sedation using midazolam administration. But, We will have to research about its optimal dosages for flumazenil, used as intranasal spray for reversal agents against the midazolam by evaluating the blood plasma concentration of midazolam and flumazenil.
This study was conducted to examine the effects of electric stimulation conditions on in vitro developmental ability of caprine embryos after somatic cell nuclear transfer. Recipient oocytes were surgically collected after superovulation by using CIDR and FSH, PMSG, hCG and estrous synchronization in Korean native goats. The caprine ear cells were cultured in vitro in serum-starvation condition (TCM-l99 + 0.5% FBS) for 3 to 5 days of cell confluence. The zona pellucida of in vivo and in vitro matured oocytes were partially drilled using laser system. Single somatic cell was individually transferred into the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3M mannitol. After the electofusion, embryos were activated by electric stimulation or Ionomycin + 6-DMAP. Nuclear transfer embryos were cultured in mSOF medium supplemented with 0.8% BSA 6∼7 days at 39 , 5% $CO_2$, 5% $O_2$, 90% $N_2$. The fusion rate of donor cells was 60.4% and 40.3 % in ear cell and fetal fibroblast, and cleavage rate were 40.6% and 48.2%, respectively. No significant difference was found in the fusion and cleavage rate in different donor cells. Nuclear transferred oocytes were fused by electric pulses of 1.30∼1.40, 2.30∼2.39 and 2.40∼2.46 ㎸/cm. There was no significant difference among different electric pulses in fusion rates (26.7, 34.8 and 43.8%). The cleavage rate was higher (p<0.05) in 1.30∼1.40 ㎸/cm (82.9%) than 2.30∼2.39 ㎸/cm (43.8%) and 2.40∼2.46 ㎸/cm. (51.8%). The fusion rates of recipient oocyte source were 1st (43.5% and 23.6%), 2nd (55.7% and 39.2%) and 3rd (66.1% and 52.8%) in in vivo and in vitro oocytes. However, fusion ratee were significantly higher (p<0.05) in in vivo than in vitro oocyte. The cleavage rate of fused oocytes from in vivo and in vitro sources were 52.6% and 54.4%, respectively. No significant difference was found in the cleavage rate according to the recipient oocyte source. These results suggest that factors such as field pulse of electric stimulation and oocyte source could affect in vitro developmental ability of nuclear transplanted caprine oocytes.
Kim, Se-Woong;Kim, Dae-Hwan;Jung, Yeon-Gil;Roh, Sang-Ho
Reproductive and Developmental Biology
/
v.35
no.1
/
pp.17-22
/
2011
This study was designed to determine the effect of electric field strength, duration and fusion buffer in fusion parameters on the rate of membrane fusion between the somatic cell and cytoplast for Korean cattle (HanWoo) somatic cell nuclear transfer (SCNT) procedure. Following electrofusion, effect of 5 or $10\;{\mu}M$$Ca^{2+}$-ionophore of activation treatment on subsequent development was also evaluated. Cell fusion rates were significantly increased from 23.1% at 20 V/mm to 59.7% at 26 V/mm and 52.9% at 27 V/mm (p<0.05). Due to higher cytoplasmic membrane rupture or cellular lysis, overall efficiency was decreased when the strength was increased to 30 V/mm (18.5%) and 40 V/mm (6.3%) and the fusion rate was also decreased when the strength was at 25 V/mm or below. The optimal duration of electric stimulation was significantly higher in $25\;{\mu}s$ than 20 and $30\;{\mu}s$ (18.5% versus 9.3% and 6.3%, respectively, p<0.05). Two nonelectrolyte fusion buffers, Zimmermann's (0.28 M sucrose) and 0.28 M mannitol solution for cell fusion, were used for donor cell and ooplast fusion and the fusion rate was significantly higher in Zimmermann's cell fusion buffer than in 0.28 M mannitol (91.1% versus 48.4%, respectively, p<0.05). The cleavage and blastocyst formation rates of SCNT bovine embryos activated by $5\;{\mu}M$$Ca^{2+}$-ionophore was significantly higher than the rates of the embryos activated with $10\;{\mu}M$ of $Ca^{2+}$-ionophore (70.0% versus 42.9% and 22.5% versus 14.3%, respectively; p<0.05). This result is the reverse to that of parthenotes which shows significantly higher cleavage and blastocyst rates in $10\;{\mu}M$$Ca^{2+}$-ionophore than $5\;{\mu}M$ counterpart (65.6% versus 40.3% and 19.5% versus 9.7%, respectively; p<0.05). In conclusion, SCNT couplet fusion by single pulse of 26 V/mm for $25\;{\mu}s$ in Zimmermann's fusion buffer followed by artificial activation with $5\;{\mu}M$$Ca^{2+}$-ionophore are suggested as optimal fusion and activation methods in Korean cattle SCNT protocol.
Kim, Dongshin;Koo, Yong-Sung;Kim, Ju-Hee;Kang, Soyeon;Oh, Wonwook;Chan, Sung-Il
Journal of the Korea Academia-Industrial cooperation Society
/
v.18
no.6
/
pp.37-43
/
2017
This review investigates the basic principle of physical interactions and failure mechanisms introduced in the materials and inner parts of semiconducting components under electromagnetic pulses (EMPs). The transfer process of EMPs at the semiconducting component level can be explained based on three layer structures (air, dielectric, and conductor layers). The theoretically absorbed energy can be predicted by the complex reflection coefficient. The main failure mechanisms of semiconductor components are also described based on the Joule heating energy generated by the coupling between materials and the applied EMPs. Breakdown of the P-N junction, burnout of the circuit pattern in the semiconductor chip, and damage to connecting wires between the lead frame and semiconducting chips can result from dielectric heating and eddy current loss due to electric and magnetic fields. To summarize, the EMPs transferred to the semiconductor components interact with the chip material in a semiconductor, and dipolar polarization and ionic conduction happen at the same time. Destruction of the P-N junction can result from excessive reverse voltage. Further EMP research at the semiconducting component level is needed to improve the reliability and susceptibility of electric and electronic systems.
Laser wave length can have evaporation effect by absorption because outer skin or tissue of focus is consisted of water almost though absorption of water occurs more than 90% almost in formation thickness of very thin floor. Can operate outer skin, steam by floor and correct incision of formation is available. Suture surgical operation is available to vein or lymph system and surgical operation region can dry and see as eye and radish bleeding surgical operation is available. Specially, stability of tube both end output about pulse by weight very, this research can cause various curative effect because can reduce bulk and control easily current wave style of medical laser using electric power conversion device of high frequency way. If introduce ZVS (Zero Voltage Switching) or ZVZCS (Zero Voltage and Zero Current Switching), is more profitable because can reduce switching damage. Because electric power department of proposed medical laser can do stable soft-switching in wide subordinate extent introducing ZVZCS technique by the first help and control department composes microcontroller, output current waveform user have free form make Result that experiment because design and manufacture, brought result that improve of 20% than existing equipment, and will be bought to get into superior result if supplement as systematic late.
Kim, Hoe-Geun;Song, Myeon-Gyu;Kim, Dong-U;Lee, Sang-Yul
Proceedings of the Korean Institute of Surface Engineering Conference
/
2018.06a
/
pp.62-62
/
2018
Pt is still considered as one of the most active electrocatalysts for ORR in alkaline fuel cells. However, the high cost and scarcity of Pt hamper the widespread commercialization of fuel cells. As a strong candidate for the replacement of Pt catalyst, silver (Ag) has been extensively studied due to its high activity, abundance, and low cost. Ag is more stable than Pt in the pH range of 8~14 as the equilibrium potential of Ag/Ag+ being ${\approx}200mV$ higher than that of Pt/PtO. However, Ag is the overall catalytic activity of Ag for oxygen reduction reaction(ORR) is still not comparable to Pt catalyst since the surface Ag atoms are approximately 10 times less active than Pt atoms. Therefore, further enhancement in the ORR activity of Ag catalysts is necessary to be competitive with current cutting-edge Pt-based catalysts. We demonstrate the architectural design of Ag catalysts, synthesized using plasma discharge in liquid phase, for enhanced ORR kinetics in alkaline media. An attractive feature of this work is that the plasma status controlled via electric-field could form the Ag nanowires or dendrites without any chemical agents. The plasma reactor was made of a Teflon vessel with an inner diameter of 80 mm and a height of 80 mm, where a pair of tungsten(W) electrodes with a diameter of 2 mm was placed horizontally. The stock solutions were made by dissolving the 5-mM AgNO3 in DI water. For the synthesis of Agnanowires, the electricfield of 3.6kVcm-1 in a 200-ml AgNO3 aqueous solution was applied across the electrodes using a bipolar pulsed power supply(Kurita, Seisakusyo Co. Ltd). The repetition rate and pulse width were fixed at 30kHz and 2.0 us, respectively. The plasma discharge was carried out for a fixed reaction time of 60 min. In case of Ag nanodendrites, the electric field of 32kVcm-1 in a 200-ml AgNO3 aqueous solution was applied and other conditions were identical to the plasma discharge in water in terms of electrode configuration, repetition rate and discharge time. Using SEM and STEM, morphology of Ag nanowires and dendrites were investigated. With 3.6 kV/cm, Ag nanowire was obtained, while Ag dendrite was constructed with 32 kV/cm. The average diameter and legth of Ag nanowireses were 50 nm and 3.5 um, and thoes values of Ag dendrites were 40 nm and 3.0 um. As a results of XPS analysis, the surface defects in the Ag nanowires facilitated O2 incorporation into the surface region via the interaction between the oxygen and the electron cloud of the adjacent Ag atoms. The catalytic activity of Ag for oxygen reduction reaction(ORR) showed that the catalytic ORR activity of Ag nanowires are much better than Ag nanodendrites, and electron transfer number of Ag nanowires is similar to that of Pt (${\approx}4$).
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