• Title/Summary/Keyword: clamp

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Characteristics of the inward current and its changes following fertilization in hamster eggs (햄스터 난자에서 관찰되는 내향전류의 성상과 수정후의 변화)

  • Han, Jae-hee;Hong, Seong-geun
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.280-289
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    • 1998
  • Voltage-sensitive ion channels contribute to establishment of the cell excitablity and the generation of the cellular function. At hamster oocytes in the primitive stage during developing process, an inward current elicited by voltage pulses was found to be carried mainly by $Ca^{2+}$. Even at present, $Ca^{2+}$ channels serve as the most probable route to pass this inward current but there is no evidence of the presence of this channels in eggs. To date, both the characteristic properties and the physiological role in the early stage of development remain unclear. Here we examined the characteristic properties of the inward current and changes in this currents at unfertilized oocytes, fertilized zygotes and two-cell embryos using whole-cell voltage clamp technique. The inward current carried reportedly by $Ca^{2+}$ was remained following removing external $Ca^{2+}$ but completely abolished by further replacement of impermeants such as tetramethylammonium ion ($TMA^+$) or $choline^+$ instead of $[Na^+]_0$. Tetrodotoxin did not affect on this inward current remained at $[Ca^{2+}]_0$-free condition. Removal of $Na^+$ ion out of the experimental solution clearly decreased the current. After adding 2mM $Ca^{2+}$ to the $Na^+$-free media, the inward current was restored. Interestingly, this current carried by either $Ca^{2+}$ or $Na^+$ was decreased by the reduction of intracellular $Cl^-$ concentration, or by $Cl^-$ channel blockers such as niflumic acid, DIDS and SITS. When $Cl^-$ concentration was lowered without changes in other ionic components, this inward current was reduced. At fertilized oocytes and two-cell embryos, the inward current carried by $Ca^{2+}$ and $Na^+$ was severely reduced. Also $Cl^-$ component could not be observed. From these results, the inward current is composed of $Ca^{2+}$, $Na^+$ and $Cl^-$ component, suggesting that the channel carrying this inward current is not selective specifically to $Ca^{2+}$. During early stage of development, the voltage-sensitive ion current seems not to contribute essentially to the cell cleavage and differentiation. The loss of $Cl^-$ component after fertilization suggests that $Cl^-$ may play a role in maintaining the viability of unfertilized ova.

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Off-Pump Coronary Artery Bypass Grafting (심폐바이패스없이 시행하는 관상동맥우회술)

  • Kim, Ki-Bong;Lim, Hong-Gook;Huh, Jae-Hak;Ahn, Hyuk;Ham, Byung-Moon
    • Journal of Chest Surgery
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    • v.33 no.1
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    • pp.38-44
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    • 2000
  • Background: We analyzed the result of the "Off-Pump" Coronary Artery Bypass grafting (OPCAB) performed to minimize inflammatory responses to cardiopulmonary bypass and myocardial ischemia during the aortic cross-clamp period. Material and Method : The preoperative diagnosis operative procedure mortality complication and postoperative course of the 50 patients who underwent OPCAB between January 1998 and September 1998 were analyzed. There were 34 males and 16 females with mean age of 60$\pm$9 years. Preoperative clinical diagnoses were unstable angina in 31(62%) stable angina in 16(32%) and clinical diagnoses were unstable angina in 31(62%) stable angina in 16(32%) and postinfarction angina in 3(6%) patients. Preoperative angiographic diagnoses were three-vessel disease in 25(50%) two-vessel disease in 5(10%) one-vessel disease in 7(14%) and left main disease in 13(26%) patients. There were elective operation in 37 cases and urgent operation in 13 cases. Result: The mean number of grafts was 3.2$\pm$1.2 per patient. Grafts used were unilateral internal thoracic artery in 43 greater saphenous vein in 37 radial artery in 7 bilateral internal thoracic arteries in 4 and right gastroepiploic artery in 2 cases Forty sequential anastomoses were performed in 18 cases. Vessels accessed were left anterior descending artery in 48 diagonal branch in 41 obtuse marginal branch in 30 right coronary artery in 24 posterior descending artery in 9 ramus intermedius in 5 and posterolateral branch in 5 anastomoses. Predischarge coronary angiography performed in 44 patients demonstrated the patency rate of 89.5%(128/143) Operative mortality was 2%(1/150) Postoperative complications were arrhythmia in 5 graft occlusion that needed reoperation in 4. perioperative myocardial infarction in 2 femoral artery thromboembolism developed after the application of IABP in 1 postoperative transient delirium in 1 peripheral compression neuropathy in 1 case. Sixteen patients(32%) were extubated at the operating room and the other patients were extubated at the mean 13$\pm$20 hours after the operation. Mean duration of stay in intensive care unit was 49$\pm$46 hours. Thirteen patients(26%) required blood transfusions perioperatively and the amount of perioperative blood transfusion was mean 0.70$\pm$1.36 pack/patient. Conclusion: OPCAB is suggested to be the ideal technique with less postoperative complication less hospitalization time and less cost.less cost.

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Four Voltage-Gated Potassium Currents in Trigeminal Root Ganglion Neurons

  • Choi, Seung Ho;Youn, Chang;Park, Ji-Il;Jeong, Soon-Yeon;Oh, Won-Man;Jung, Ji-Yeon;Kim, Won-Jae
    • International Journal of Oral Biology
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    • v.38 no.1
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    • pp.13-19
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    • 2013
  • Various voltage-gated $K^+$ currents were recently described in dorsal root ganglion (DRG) neurons. However, the characterization and diversity of voltage-gated $K^+$ currents have not been well studied in trigeminal root ganglion (TRG) neurons, which are similar to the DRG neurons in terms of physiological roles and anatomy. This study was aimed to investigate the characteristics and diversity of voltage-gated $K^+$ currents in acutely isolated TRG neurons of rat using whole cell patch clamp techniques. The first type (type I) had a rapid, transient outward current ($I_A$) with the largest current size having a slow inactivation rate and a sustained delayed rectifier outward current ($I_K$) that was small in size having a fast inactivation rate. The $I_A$ currents of this type were mostly blocked by TEA and 4-AP, K channel blockers whereas the $I_K$ current was inhibited by TEA but not by 4-AP. The second type had a large $I_A$ current with a slow inactivation rate and a medium size-sustained delayed $I_K$ current with a slow inactivation rate. In this second type (type II), the sensitivities of the $I_A$ or $I_K$ current by TEA and 4-AP were similar to those of the type I. The third type (type III) had a medium sized $I_A$ current with a fast inactivation rate and a large sustained $I_K$ current with the slow inactivation rate. In type III current, TEA decreased both $I_A$ and $I_K$ but 4-AP only blocked $I_A$ current. The fourth type (type IV) had a smallest $I_A$ with a fast inactivation rate and a large $I_K$ current with a slow inactivation rate. TEA or 4-AP similarly decreased the $I_A$ but the $I_K$ was only blocked by 4-AP. These findings suggest that at least four different voltage-gated $K^+$ currents in biophysical and pharmacological properties exist in the TRG neurons of rats.

Selection of parental monokaryons from Korean Hypsizigus marmoreus by protoplast regeneration (원형질체 재생을 통한 느티만가닥버섯 단핵균주 선발)

  • Oh, Youn-Lee;Kong, Won-Sik;Jang, Kab-Yeul;Shin, Pyung-Gyun;Kim, Eun-Sun;Oh, Min ji;Choi, In-Geol
    • Journal of Mushroom
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    • v.13 no.3
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    • pp.270-273
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    • 2015
  • Hypsizigus marmoreus is commercially the most important edible mushroom in Japan. This mushroom is usually cultivated for a longer period (about 85~120 days) than other mushroom. In order to develop a new cultivar that has a shortened cultivation period, the genome analysis of this strain has been considered. This study aims to obtain parental monokaryotic strains reproducing 'Haemi' cultivar in Hypsizigus marmoreus for reference genome sequencing. The mycelia were cultured in MCM and MYG media for various incubation periods. Homogenized mycelia were treated with commercial cell wall degrading enzymes to maximize protoplasts production yield from Hypsizigus marmoreus. The greatest number of protoplasts was obtained from mycelia cultured in MCM media for 3 days using Novozyme enzyme. The isolated protoplasts were grown in regeneration agar media after two weeks. Regenerated colonies were picked and moved on separated dishes for microscopic observation. Neohaplonts regenerated from dikayotic strains were identified by the absence of clamp connections. We confirmed that one of monokaryotic strains is a parental strain by crossing with an original compatible strain of 'Haemi' cultivar. This parental strain will be used for reference genome sequence analysis.

Effects of [D-$Pen^2$, D-$Pen^5$]-enkephalin on the Neuronal Activity of Medial Vestibular Nuclear Neurons

  • Jang, Su-Jeong;Jeong, Han-Seong;Park, Jong-Seong
    • Biomedical Science Letters
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    • v.15 no.3
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    • pp.199-205
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    • 2009
  • This study was designed to investigate direct effects of [D-$Pen^2$, D-$Pen^5$]-enkephalin, a $\delta$-opioid receptor agonist on the neuronal activity of medial vestibular nuclear (MVN) neurons by whole-cell configuration patch clamp experiments. The spike frequency of MVN neuron was increased to $9.50{\pm}0.55$ (P<0.05) and $10.56{\pm}0.66$ (P<0.05) by 5 and $10{\mu}M$ [D-$Pen^2$, D-$Pen^5$]-enkephalin from the control level of $8.05{\pm}0.55$ spikes/sec, respectively (n=18). The resting membrane potential of the neurons was increased to $-37.86{\pm}0.92$ and $-36.97{\pm}0.97$ (P<0.05) from $-38.74{\pm}1.13\;mV$ by 5 and $10{\mu}M$ [D-$Pen^2$, D-$Pen^5$]-enkephalin, respectively. The amplitude of afterhyperpolarization was decreased to $23.78{\pm}0.65$ and $21.67{\pm}0.89$ (P<0.05) from $23.73{\pm}0.53\;mV$ by 5 and $10{\mu}M$ [D-$Pen^2$, D-$Pen^5$]-enkephalin, respectively. The spike width was changed to $2.22{\pm}0.08$ and $2.24{\pm}0.07$ from $2.20{\pm}0.08\;mV$ by 5 and $10{\mu}M$ [D-$Pen^2$, D-$Pen^5$]-enkephalin, respectively. After pretreatment of naltrindole, a highly selective 8-opioid receptor antagonist, [D-$Pen^2$, D-$Pen^5$]-enkephalin did not change firing rate, resting membrane potential, afterhyperpolarization amplitude, and spike width of MVN neurons. The above experimental results suggest that [D-$Pen^2$, D-$Pen^5$]-enkephalin increases the neuronal activity of MVN neurons via inhibition of calcium-dependent potassium currents underlying the afterhyperpolarization.

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Analysis of antigenic domain of GST fused major surface protein (p30) fragments of Toxoplasma gondii (융합단백질로 발현된 톡소포자충의 주요막단백질(p30) 절편의 항원성)

  • 남호우;임경심
    • Parasites, Hosts and Diseases
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    • v.34 no.2
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    • pp.135-142
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    • 1996
  • Antigenic domain of jai or surface protein (p30) of Toxoplosmc Sondii was analyzed after polymerase chain reaction (PCR) of its gene fragments. Hydrophilic or hydrophobic moiety of amino acid sequences were expressed as glutathione S-transferase (G57) fusion proteins. Fragments of p30 gene were as follows: 737, total p30 open reading frame (ORF) ; S28, total ORF excluding N-terminal signal sequence and C-terminal hydrophobic sequence; Al9, N-terminal 2/3 parts of A28; A19, N-terminal 2/3 of S28; P9, C-terminal 2/3 part of S28; Z9. middle 1/3 of S28; and 29, C-terminal 1/3 of S28. respectively. Primer of each fragment was synthesized to include clamp sequence of EcoR I restriction site. PCR amplified DNA was inserted info GST (26 kDa) expression vector, PGEX-47-1 to transform into Escheri,hia coei (.JM105 strain). G57 fusion proteins were expressed with IPTG induction as 63. 54, 45, 45, 35, 36. and 35 kDa proteins measured by SDS-PAGE. Each fusion protein was confirmed with G57 detection kit. Western blot analysis with the serum of a toxoplasmosis patient revealed antigenicity in proteins expressed by T37. S28, and Al9 but not those by Pl8. X9, Y10, and Z9. Antigenicity of p30 seems to be located either in N-terminal 115 part in the presence of middle 1/3 part or in the oligopeptides between margins of the first and second 1/3 parts.

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The Design of SCR-based Whole-Chip ESD Protection with Dual-Direction and High Holding Voltage (양 방향성과 높은 홀딩전압을 갖는 사이리스터 기반 Whole-Chip ESD 보호회로)

  • Song, Bo-Bae;Han, Jung-Woo;Nam, Jong-Ho;Choi, Yong-Nam;Koo, Yong-Seo
    • Journal of IKEEE
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    • v.17 no.3
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    • pp.378-384
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    • 2013
  • We have investigated the electrical characteristics of SCR(Silicon Controlled Rectifier)-based ESD power clamp circuit with high holding voltage and dual-directional ESD protection cells for a whole-chip ESD protection. The measurement results indicate that the dimension of n/p-well and p-drift has a great effect on holding voltage (2V-5V). Also A dual-directional ESD protection circuit is designed for I/O ESD protection application. The trigger voltage and the holding voltage are measured to 5V and 3V respectively. In comparison with typical ESD protection schemes for whole-chip ESD protection, this ESD protection device can provide an effective protection for ICs against ESD pulses in the two opposite directions, so this design scheme for whole-chip ESD protection can be discharged in ESD-stress mode (PD, ND, PS, NS) as well as VDD-VSS mode. Finally, a whole-chip ESD protection can be applied to 2.5~3.3V VDD applications. The robustness of the novel ESD protection cells are measured to HBM 8kV and MM 400V.

Titanium Hemostatic Clip Tailoring Method to Overcome Vessel Caliber Discrepancy in Interposition Saphenous Vein Graft for Carotid Artery Resection (경동맥 절제술 후 재건에 사용된 복재정맥의 혈관구경 불일치를 극복하기 위한 Titanium Hemostatic Clip 봉합방법)

  • Kim Sun-Ho;Lim Young-Chang;Lee Sei-Young;Lim Jae-Yol;Choi Eun-Chang
    • Korean Journal of Head & Neck Oncology
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    • v.20 no.2
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    • pp.177-180
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    • 2004
  • Objectives: To completely excise a malignant tumor which invades carotid artery walls, the resection and reconstruction of the carotid artery is essentially required. In most of the cases, interposition graft using a saphenous vein has been performed, however the vessel caliber discrepancy between a common carotid artery and the saphenous vein can result in a problem on surgical technique. We have introduced and evaluated a new titanium hemoclip tailing method to overcome vessel caliber discrepancy in interposition saphenous vein graft for carotid artery resection in the treatment of head and neck cancers. Method: After carotid artery resection, the calibers of the proximal common carotid artery and the vein were compared, and an orifice of the common carotid artery was gradually reduced to a little larger than or the same size as the orifice of the vein by using the titanium hemostatic clip. Subsequently, the common carotid artery was connected to the vein through anastomosis. The same method was also applied to the distal anastomosis site. There after, the vessels were connected through the anastomosis, and a circulation was restored by releasing a vascular clamp. Then, a titanium hemostatic clip-applied redundant portion on the outside of carotid artery was sutured by the blanket edge suture method, using 6-0 Prolene. Results: We have experienced this method in two patients with recurrent squamous cell carcinoma and neuroblostoma, respectively. The interposition saphenous vein graft of these patients was found to maintain good patency on the follow up angiography after one year, and they had no specific vascular complication, such as atherosclerosis. Conclusion: This method made it possible to simply perform the interposition saphenous vein graft (ISVG) within a short time and, therefore, was very useful for shortening the duration to block circulation.

An Experience of Cox-maze III Procedure for Chronic Atrial Fibrillation (만성 심방세동에 대한 Cox-maze III 수술의 임상경험)

  • 김삼현;박이태;서필원;박성식;류재욱;최창휴;김명아;이명용;김영권
    • Journal of Chest Surgery
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    • v.31 no.7
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    • pp.668-673
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    • 1998
  • During the past several years, the maze operation has become the most effective method of treatment for chronic atrial fibrillation. When the maze procedure is done concomittantly with other cardiac operations, surgeons, in their initial experiences, may be concerned about the additional operative risks and uncertainty of the results. We performed the Cox-maze III procedure in six cases of chronic atrial fibrillation associated with mitral, mitral & aortic, or coronary arterial disease. Maze III procedure was done with open mitral commissurotomy(3 cases), mitral valve replacement(1 case), aortic and mitral valve replacement(1 case), and two-vessel coronary bypass graft(1 case). In spite of rather prolonged aortic cross clamp time, cardiac recovery was uneventful in all cases. No cases required reexploration for postoperative bleeding. All patients showed regular sinus rhythms immediate or between 2 and 20 days postoperateratively. Transient postoperative supraventricular arrhythmarias were easily controlled by various antiarrhythmic agents. In follow up evaluations, all cases showed regular sinus rhythm on ECG and the right and left atrial transport function was confirmed by Doppler echocardiography in all except one. Though our experience was limited in case number, the Cox-maze III procedure was effective in controlling the chronic atrial fibrillation without serious additional operative risks.

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Design of 20 W Class-E Amplifier Including Protection for Wireless Power Transmission at ISM 13.56 MHz (보호 회로를 포함한 무선 전력 전송용 ISM 13.56 MHz 20 W Class-E 앰프 설계)

  • Nam, Min-Young;Kim, Young-Sik
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.24 no.6
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    • pp.613-622
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    • 2013
  • In this paper, an inductive clamping class-E power amplifier has been tested for wireless power transmission at ISM band, 13.56 MHz. The implemented power amplifier is designed to operate stably without destroying power transistor in wireless power transmission system which basically keeps not to align between a transmitting antenna and a receiving antenna. The power amplifier is also designed to enhance harmonic filtering characteristic. The amplifier was tested with a DC supply voltage of 28 V and input power of 25 dBm at 13.56 MHz. The test results show the output power level of 43 dBm, the difference power level between fundamental frequency and second harmonic frequency of more than 55 dBc, the dc current consumption of 830 mA, and the high power-added efficiency of 85 %. Finally, the implemented power amplifier operated normally with 830 mA DC current consumption from 28 V source when the two antennas were aligned, and the power transmission was successful. But when the two antennas were not aligned, its DC current consumption automatically decreased down to 420 mA to protect the switching transistor.