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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.

Effect of Paraspinal Muscle Fatigue on Proprioception and Coordination of the Lower Limb (부척추근의 근피로가 하지의 고유수용성 감각과 협응 능력에 미치는 영향)

  • Park, Min-Chull;Kim, Hyoung-Su;Lee, Sang-Youl;Lee, Myoung-Hee;Goo, Bong-Oh;Yoon, Chang-G
    • Journal of the Korean Society of Physical Medicine
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    • v.3 no.4
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    • pp.255-260
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    • 2008
  • Purpose : This study was performed to find out the influence of the change on proprioception and coordination of the lower limb by experimentally induced paraspinal muscle fatigue, Methods : Twelve subjects(6 males, 6 females) with no history low back and lower limbs problem volunteered for this study. Subjects were pre-tested proprioception and coordination using the MR-FSS(Mornitored Rehab Functional Squat System). And then the subjects performed isometric paraspinal muscles contraction for induced muscle fatigue. Muscle fatigue was defined surface EMG(MP 150). After isometric contraction, the subjects were post-tested using the MR-FSS. Results : In comparison of coordination, deviation of concentric coordination was increased(from $0.50{\pm}0.13cm$ to $0.67{\pm}0.21$)(p<0.05) and deviation of eccentric coordination was also increased(from $0.51{\pm}0.15cm$ to $0.70{\pm}0.26cm$)(p<0.05). And deviation movement of proprioceptive test was increased(from $4.98{\pm}1.80cm$ to $5.69{\pm}1.98cm$), but there was no significant different(p>0.05). Conclusion : Fatigue of paraspinal muscles was decreased concentric and eccentric coordination of the lower limb.

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Effect of Electrode Placement on Procaine Iontophoresis (전극배치가 Procaine 이온도입에 미치는 영향)

  • Lee, Jae-Hyoung;Song, In-Young;Kook, Jee-Youn
    • Journal of Korean Physical Therapy Science
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    • v.4 no.2
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    • pp.399-403
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    • 1997
  • The purpose of this study was to determine the efficacy of electrode placement on procaine iontophoresis. Thirty-three healthy students with an age range of 19 to 34 years, were participated in this study. The subjects were randomly assigned into 3 groups. Each subjects received iontophoresis on the proximal 1/3 of volar surface of dominant forearm with soft cotton pad($3.5{\times}3.5cm$) soaked in 2 ml of 4% procaine hydrochloride (pH 5.1) at 4 mA for 10 minutes(total current 40 mA min) of anodal DC. In transversal electrode placement(TEP) group, dispersive electrode was placed on the proximal 1/3 of dorsal surface of the forearm. In longitudinal electrode placement (LEP) group and control group, dispersive electrode were placed on the distal 1/3 of volar surface of the forearm. After procaine iontophoresis, duration of anesthesia were evaluated at five minutes intervals on five random locations in the iontophoretically area using a 21-gauge sterile hypodermic needle pressed with 1 mm invagination until return the sharp pin-pricking pain sensation. The data were ana lysed with one-way ANOVA to determine signific~nt differences between groups. The results showed significantly differences in the local anesthetic duration between the 3 groups(p<0.001). The anesthetic durations of TEP group and LEP group were significantly longer when compared with control group(p<0.05). Anesthetic durations of TEP group and LEP group were not significantly difference, but anesthetic duration of LEP group tends to longer than TEP group. In view of these results, clinicians should consider the electrode placement method when performing the iontophoresis.

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Fabrications and Properties of Al/$VF_2$/$n^+$-Si(100) Structures by Dip Coating Methode (Dip Coating 법에 의한 Al/$VF_2$-TrFE/Si(100) 구조의 제작 특성)

  • Kim, Ka-Lam;Jeong, Sang-Hyun;Yun, Hyeong-Seon;Lee, Woo-Seok;Kwak, No-Won;Kim, Kwang-Ho
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.11a
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    • pp.20-21
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    • 2008
  • Ferroelectric vinylidene fluoride-trifluoroethylene ($VF_2$-TrFE) copolymer films were directly deposited on degenerated Si ($n^+$, 0.002 $\Omega{\cdot}cm$) using by dip coating method. A 1 ~ 3 wt% diluted solution of purified vinylidene fluoride-trifluoroethylene ($VF_2$:TrFE=70:30) in a dimethylformamide (DMF) solvent were prepared and deposited on silicon wafers using dip coating method for 10 seconds. After Post-Annealing in a vacuum ambient at 100~200 $^{\circ}C$ for 60 min, upper aluminum electrodes were deposited by thermal evaporation through the shadow mask to complete the MFS structure. The ferroelectric $\beta$-phase peak of films, depending on the annealing temperature, started to show up around $125^{\circ}C$, and the intensity of the peak increased with increasing annealing temperature. Above $175^{\circ}C$, the peak started to decrease. The C-V characteristics were measured using a Precision LCR meter (HP 4284A) with frequency of 1MHz and a signal amplitude of 20 mV. The leakage-current versus electric-field characteristics was measured by mean of a pA meter/DC voltage source (HP 4140B).

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Surface Characteristics of Anodized and Hydrothermally-Treated Ti-6Al-7Nb Alloy (양극산화와 열수처리한 Ti-6Al-7Nb 합금의 표면 특성)

  • Kim, Moon-Young;Song, Kwang-Yeob;Bae, Tae-Sung
    • Journal of Dental Rehabilitation and Applied Science
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    • v.21 no.1
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    • pp.33-42
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    • 2005
  • This study was performed to investigate the surface properties and in vitro biocompatibility of electrochemically oxidized Ti-6Al-7Nb alloy by anodic spark discharge technique. Discs of Ti-6Al-7Nb alloy of 20 mm in diameter and 2 mm in thickness were polished sequentially from #300 to 1000 SiC paper, ultrasonically washed with acetone and distilled water for 5 min, and dried in an oven at $50^{\circ}C$ for 24 hours. Anodizing was performed using a regulated DC power supply. The applied voltages were given at 240, 280, 320, and 360 V and current density of $30mA/cm^2$. Hydrothermal treatment was conducted by high pressure steam at $300^{\circ}C$ for 2 hours using a autoclave. Samples were soaked in the Hanks' solution with pH 7.4 at $36.5^{\circ}C$ during 30 days. The results obtained were summarized as follows; 1. The oxide films were porous with pore size of $1{\sim}5{\mu}m$. The size of micropores increased with increasing the spark forming voltage. 2. The main crystal structure of the anodic oxide film was anatase type as analyzed with thin-film X-ray diffractometery. 3. Needle-like hydroxyapatite (HA) crystals were observed on anodic oxide films after hydrothermal treatment at $300^{\circ}C$ for 2 hours. The precipitation of HA crystals was accelerated with increasing the spark forming voltage. 4. The precipitation of the fine asperity-like HA crystals were observed after being immersed in Hanks' solution at $37^{\circ}C$. The precipitation of HA crystals was accelerated with increasing the spark forming voltage and the time of immersion in Hanks' solution. 5. The Ca/P ratio of the precipitated HA layer was equivalent to that of HA crystal as increasing the spark forming voltage and the time of immersion in Hanks' solution.

Theoretical considerations on the giant magnetoimpedance effect in amorphous ribbons

  • Phan, Manh-Huong;Nguyen Cuong;Yu, Seong-Cho
    • Proceedings of the Korean Magnestics Society Conference
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    • 2003.06a
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    • pp.60-61
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    • 2003
  • Theoretical considerations on a giant magneto-impedance (GMI) effect in amorphous ribbons (i.e., thin films) have been made in terms of the expressions of effective permeability and impedance derived in the frame of classical electrodynamics and ferromagnetism. The dependence of GMI effect on the external do magnetic field (H$\_$ext/) and the frequency of alternating current are simulated and discussed in the knowledge of energy conversion consisting of the current energy loss, the ferromagnetic energy consumption, and the magnetic energy storage in the film. The obtained results are summarized as follow: (a) As frequency f< 20 ㎒, the real part of effective permeability (${\mu}$′) changes slightly. The peak of the ${\mu}$′curve always locates at H$\_$ext/=H$\_$ani/ - the anisotropy field. However, the peak value of ${\mu}$′ tends to increase with increasing frequency in the frequency range of 11-20 ㎒. (b) In the frequency range, f= 21-23 ㎒, a negative peak additionally appears. Meanwhile, both the positive and negative peak values rapidly increase with increasing frequency and their peak positions shift towards a high H$\_$ext/. (c) The positive peak value of ${\mu}$′ starts to decrease at f= 29 ㎒ and its negative peak does so at about 35 ㎒. Then, both peaks keep such a tendency and their peak positions move to high H$\_$ext/, as increasing frequency. (d) The dependence of the imaginary part of effective permeability (${\mu}$") on the external dc magnetic field and the frequency of the alternating field indicates that there is only one peak involved in ${\mu}$" for the whole frequency range. (e) The impedance vs. magnetic field curves at various frequencies show that there is a critical value of frequency around f= 18-19 ㎒ where the transition between two frequency regimes occurs; the one (low frequency) in which ${\mu}$′ predominantly contributes to the GMI effect and the other (high frequency) in which ${\mu}$" determines the GMI effect.

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Formation and Hydrogen Absorption Properties of Intermetallic Mg-Ni Compound Nanoparticles (Mg-Ni 금속 간 화합물 나노입자의 형성과 수소저장 특성)

  • BAE, YOOGEUN;HWANG, CHULMIN;KIM, JONGSOO;DONG, XING LONG;KIM, SEWOONG;JUNG, YOUNGUAN
    • Journal of Hydrogen and New Energy
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    • v.28 no.3
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    • pp.238-245
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    • 2017
  • Mg-Ni nanoparticles were synthesized by a physical vapor condensation method (DC arc-discharge) in a mixture of argon and hydrogen atmosphere, using compressed mixture of micro powders as the raw materials. The crystal phases, morphology, and microstructures of nanoparticles were analyzed by means of X-ray diffraction (XRD) and high-resolution transmission electron microscopy (HRTEM). It was found that the intermetallic compounds of $Mg_2Ni$ and $Mg_2Ni$ formed with existence of phases of Mg, Ni, and MgO in Mg-Ni nanoparticles. After one cycle of hydrogen absorption/desorption process (activation treatment), Mg-Ni nanoparticles exhibited excellent hydrogen absorption properties. $Mg_2Ni$ phase became the main phase by aphase transformation during the hydrogen treatments. The phenomenon of refinement of grain size in the nanoparticle was also observed after the hydrogen absorption/desorption processes, which was attributed to the effect of volume expansion/shrinkage and subsequent break of nanoparticles. Maximum hydrogen absorption contents are 1.75, 2.21 and 2.77 wt.% at 523, 573 and 623 K, respectively.

Behavioral Characteristics Following Lesions of the Nucleus Accumbens Septi in Rats (측좌각의 부위별 파괴가 행동에 미치는 영향)

  • Lee, Soon-Chul
    • The Korean Journal of Pharmacology
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    • v.21 no.2
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    • pp.111-118
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    • 1985
  • Caudal, rostral and both areas of the nucleus accumbens septi (NAB) were lesioned each in separate animal group by applying DC of 3.0 mA for 15 sec. in order to examine if any regional differences in the NAB with regard to the manifestation of locomotor activity, The r-NAB and cr-NAB-lesioned rats were significantly increased in locomotor activity but not in the c-NAB-lesioned rats and the effect following the NAB lesion was immediately produced and returned to control levels in about 7 days postoperatively. On the other hand, the locomotor stimulation produced by methamphetamine was significantly attenuated in all NAB-lesioned rats but also stereotyped behavior was significantly elicited simultaneously. These results thus suggested that NAB may be attributed to the inhibitory role in the locomotor activity, of which intensity differs from rostral site and caudal site, and stereotyped behavior may be shown higher sensitivity of the denervated striatal dopamine function.

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Research Trends for Performance, Safety, and Comfort Evaluation of Agricultural Tractors: A Review

  • Kabir, Md. Shaha Nur;Ryu, Myong-Jin;Chung, Sun-Ok;Kim, Yong-Joo;Choi, Chang-Hyun;Hong, Soon-Jung;Sung, Je-Hoon
    • Journal of Biosystems Engineering
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    • v.39 no.1
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    • pp.21-33
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    • 2014
  • Background: Significant technological development and changes happened in the tractor industries. Contrariwise, the test procedures of the major standard development organizations (SDO's) remained unchanged or with a little modification over the years, demanding new tractor test standards or improvement of existing ones for tractor performance, safety, and comfort. Purpose: This study focuses on reviewing the research trends regarding performance, safety and comfort evaluation of agricultural tractors. Based on this review, few recommendations were proposed to revise or improve the current test standards. Review: Tractor power take-off power test using the DC electric dynamometer reduced human error in the testing process and increased the accuracy of the test results. GPS signals were used to determine acceleration and converted into torque. High capacity double extended octagonal ring dynamometer has been designed to measure drawbar forces. Numerical optimization methodology has been used to design three-point hitch. Numerous technologies, driving strategies, and transmission characteristics are being considered for reducing emissions of gaseous and particulate pollutants. Engine emission control technology standards need to be revised to meet the exhaust regulations for agricultural tractors. Finite Element Analysis (FEA) program has been used to design Roll-Over Protective Structures (ROPS). Program and methodology has been presented for testing tractor brake systems. Whole-body vibration emission levels have been found to be very dependent upon the nature of field operation performed, and the test track techniques required development/adaptation to improve their suitability during standardized assessment. Emphasizes should be given to improve visibility and thermal environment inside the cab for tractor operator. Tractors need to be evaluated under electromagnetic compatibility test conditions due to large growing of electronic devices. Research trends reviewed in this paper can be considered for possible revision or improvement of tractor performance, safety, and comfort test standards.

Effects of HT7 Acupuncture on c-Eos Expression in Limbic System of the Brain in Maternally-Separated Rats (신문혈(神門穴) 침자극(鍼刺戟)이 모성분리(母性分離) 흰쥐의 대뇌(大腦) 변연계(邊緣系)의 c-Fos 발현(發顯)에 미치는 영향(影響))

  • Lim, Sabina;Hwang, Se-Hee;Ryu, Yeun-Hee;Lee, Hye-Jung;Park, Hi-Joon
    • Korean Journal of Acupuncture
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    • v.21 no.3
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    • pp.77-88
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    • 2004
  • Acupuncture has been shown to have therapeutic effects on many mental disorders such as depression, anxiety, and substance abuse in human and animal studies. Maternal separation (MS) is a risk factor in the development of mood-related disorders such as depression. This study investigated the effects of acupuncture during MS by evaluating locomotion as behavioral markers of depression and by observing changes in c-Fos expression in the limbic system (hippocampus, amygdala and cingulate cortex) to elucidate the mechanism of the therapeutic effect. The employed acupoint, Shinmun (HT7), has been clinically used to treat mental disorders. MS for 7 days beginning on postnatal day 14 induced a significant decrease in locomotion, while acupuncture treatment at acupoint HT7 resulted in a significant increase in locomotion. c-Fos immunoreactive cells in area dentate gyrus (DG) of the hippocampus were increased in the MS group, but significantly decreased in the acupuncture group. These findings suggest that acupuncture has an effect on the depression-like disorder caused by MS, possibly by modulating c-Fos expression in DC of the hippocampus.

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