• Journal of Embryo Transfer
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• v.29 no.4
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• pp.397-401
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• 2014

The boar sperm has more lipid droplets and specialty of seminal plasma compared with other species, causing difficulties of freezing sperm and decreases for the utilization of frozen semen into the artificial insemination. However, several studies reported significant results for the recovery of sperm motility and reproductive by addition of cryoprotectants and seminal plasma after thawing. This study was designed to investigate the effects of supplementation of trehalose or glycerol in the LEY (lactose and egg yolk in BTS) solution for the conventional freezing and vitrification process. Two boars aged 16 months were used to collect semen for 2 times in a week. The samples were allotted to 3 freezing solutions (LEY + glycerol 10.5% + OEP 1.5%, LEY + trehalose 1M + OEP 1.5%, and sucrose 1.5M + trehalose 1 M + OEP 1.5%) after centrifugation at 800 g for 10 minutes. Semen was equilibrated in freezing solutions for 10 minutes and injected into plastic straws with 2~3 air bubbles to minimize freezing damages. Vitrification was performed to locate sperm in 5 cm above $LN_2$ for 5 minutes, and the conventional freezing was conducted with an automatic freezer. Motility and survival rates were measured by CASA (Computer assisted sperm an alyzing system) and FITC (Fluorescein isothiocyanate), respectively after thawing semen at $50^{\circ}C$ for 12 seconds. The results were analyzed by ANOVA with STATVIEW statistical program. The vitrificatioin solution (LEY + 10.5% glycerol + 1.5% OEP) presented higher motility (20.9%) than other solutions while the solution (LEY + 1M trehalose + 1.5% OEP) showed the lowest (motility : 5.2%). However, survival rates of vitrified sperms detected by FITC showed 1~4% live sperms in almost of dead sperms at all vitrification solutions' groups, but survival rate of freezing solution of LEY + 1M trehalose + 1.5% OEP LEY and LEY + 10.5% glycerol + 1.5% OEP were showed 49%, and 79%, respectively. There were differences (P<0.05) survival rate of conventional freezing in LEY + 10.5% glycerol + 1.5% OEP and LEY + 1M trehalose + 1.5% OEP and the remaining showed no differences. The results suggested that vitrified boar semen was not enough to be utilized for the artificial insemination, but it showed possibility to utilize for ICSI and conventional freezing with glycerol would be useful method for artificial insemination in pig while we choose the outstanding semen against tolerance to freezing damages.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.235-241
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• 2017

To preserve genetic materials, cryopreservation of the semen from live animals is the main technique to establish cryo-banking system which could be used for artificial insemination and embryo transfer. However, the population of Korean black goat (KBG) becomes to dwindle in number and is now faced genetic erosion by crossbreeding with non-native breeds in small KBG farms. In this study, simple freezing method was used to preserve frozen semen from KBG using spermatozoa of cauda epididymis (CE) and electro-stimulated semen (ES). The negative effects of seminal plasma on fresh sperm was confirmed using precipitation test of Triladyl egg yolk diluent and sperm viability after thawing was compared between CE and ES spermatozoa. When seminal plasma of fresh ES semen was washed with semen washing media (SWM), the rates of live sperm shown no significant difference between CE and ES spermatozoa before freezing. However, the survival rate of frozen/thawed CE sperm was higher than ES ($74.6{\pm}10.6%$ vs $53.8{\pm}5.2%$) with significant difference (p < 0.05). The results of longevity test on frozen/thawed sperm from CE showed healthier sperm than ES. Therefore, spermatozoa from CE could be used for cryo-banking system in KBG lines. The more studies are needed to increase survival rate of ES semen.

• Journal of the Korean Society for Aeronautical & Space Sciences
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• v.33 no.5
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• pp.1-8
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• 2005

Flows in segmented arc-heaters have been calculated for prediction of experimental operating condition or for analysis and design of arc-heater itself. Some researchers succeeded in calculating accurately inner flows of a arc-heater, but could not made mathematical models which satisfy various operating conditions for many arc-heaters. this study is forced on turbulence for the generality of mathematical model. Instead of algebraic turbulence models which are frequently used for calculating inner flow of arc-heater, two equation turbulent models are used. Prediction results agree well with experiment data and it was confirmed that $k-\varepsilon$ two equation turbulence model is appropriate for a flow in an arc heater throughout extensive numerical testing.

• Journal of the Microelectronics and Packaging Society
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• v.21 no.4
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• pp.15-24
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• 2014

Recently, a variety of printing technologies, including ink jet, gravure, and roll-to-roll (R2R) printing, has generated intensive interest in the application of flexible and wearable electronic devices. However, the actual use of printing technique is much limited because the sintering process of the printed nanoparticle inks remains as a huge practical drawback. In the fabrication of the conductive metal film, a post-sintering process is required to achieve high conductivity of the printed film. The conventional thermal sintering takes considerable sintering times, and requires high temperatures. For application to flexible devices, the sintering temperature should be as low as possible to minimize the damage of polymer substrate. Several alternative sintering methods were suggested, such as laser, halogen lamp, infrared, plasma, ohmic, microwave, and etc. Eventually, the new sintering technique should be applicable to large area, R2R, and polymer substrate as well as low cost. This article reviews progress in recent technologies for several sintering methods. The advantages and disadvantages of each technology will be reviewed. Several issues for the application in R2R process are discussed.

• Proceedings of the Korean Vacuum Society Conference
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• 2014.02a
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• pp.162-162
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• 2014

최근 들어 wearable computing에 대한 수요가 증가하면서 flexible device에 대한 연구가 활발히 진행되고 있다. 하지만, flexible device를 구현하기 위해서는 기판의 damage를 줄이기 위한 저온공정, device life-time 향상을 위한 passivation, 와이어 본딩 등 다양한 문제들이 해결 되어야 한다. 이러한 문제들 중, polymer 기판과 금속간의 접착력을 향상시키기 위해서 많은 연구자들은 기판의 표면에 adhesive layer를 도포하거나 금속잉크의 solvent를 변화시키는 등의 연구를 진행해왔다. 종래의 연구는 기존 device를 대체 할 수 있을 정도의 생산성과 polymer 기판에 대한 열 적인 손상 이 문제가 되었다. 종래의 문제를 해결하기 위하여 저온공정, in-line system이 가능한 준 준 대기압 플라즈마를 사용하였다. 본 연구에서는 금속잉크를 Ink-jet으로 jetting하여 와이어 본딩 하는 과정에서 전도성 ink의 선폭을 유지시키고 접착력을 향상하기 위하여 준 대기압 플라즈마 공정을 이용하여 이러한 문제점을 해결하고자 하였다. Polymer 기판 표면에 roughness를 만들기 위해 대략 수백 nm 크기를 갖는 graphene flake를 spray coating하여 마스크로 사용하고 준 대기압 플라즈마를 이용하여 표면을 식각 함으로써 roughness를 형성시켰다. 준 대기압 플라즈마를 발생시키기 위해 double discharge system에서 6 slm/1.5 slm (He/O2) gas composition을 하부 전극에 흘려보내고 60 kHz, 5 kV 파워를 인가하였다. 동시에 상부 전극에는 30 kHz, 5 kV 파워를 인가하여 110초 동안 표면 식각 공정을 진행하였다. Graphene flake mask가 coating되어 있는 유연기판을 산소 플라즈마 처리 한 후 물에 3초 동안 세척하여 표면에 남아있는 graphene flake를 제거하고 6 slm/0.3 slm (He/SF6)의 유량으로 주파수와 파워 모두 동일 조건으로 110초 동안 표면 처리를 하였다. Figure 1은 표면 개질 과정과 graphene flake를 mask로 사용하여 얻은 roughness 결과를 SEM을 이용하여 관찰한 결과이다. 이와 같이 실험한 결과 ink와 기판간의 접촉면적을 늘려주고 접촉 각을 조절하여 Wenzel model 을 형성 할 수 있는 표면 roughness를 생성하였고 표면의 화학적 결합을 C-F group으로 치환하여 표면의 물과 접촉각 이 $47^{\circ}$에서 $130^{\circ}$로 증가하는 것을 확인하였다.

• Journal of Embryo Transfer
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• v.28 no.1
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• pp.63-71
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• 2013

In this study, we used flow a cytometric assay to evaluate plasma membrane integrity and mitochondrial activity in post-thawed sperm that was supplemented with ginsenoside-$Rg_1$. Varying concentrations of ginsenoside-$Rg_1$ (0, 25, 50 and $100{\mu}M/ml$) were used in the extender during cryopreservation to protect the DNA of thawed sperm, thereby increasing the viability and motility rate as evaluated using a computer-assisted sperm analysis (CASA) method. The results derived from CASA were used to compare the fresh, control, and ginsenoside-$Rg_1$ groups. Sperm motility and the number of progressively motile sperm were significantly (p<0.05) higher in the $50{\mu}M/ml$ ginsenoside-Rg1 group ($61.0{\pm}4.65%$) than in the control ($46.6{\pm}7.02%$), $25{\mu}M/ml$ ($46.2{\pm}4.76%$), and $100{\mu}M/ml$ ginsenoside-$Rg_1$ ($52.0{\pm}1.90%$) groups. However, the velocity distribution of post-thawed sperm did not differ significantly. Membrane integrity and MMP staining as revealed using flow cytometry were significantly (p<0.05) higher ($91.6{\pm}0.82%$) in the $50{\mu}M/ml$ ginsenoside-$Rg_1$ group than in the other groups. Here, we report that ginsenoside-$Rg_1$ affects the motility and viability of boar spermatozoa. Moreover, ginsenoside-$Rg_1$ can be used as a protective additive for the suppression of intracellular mitochondrial oxidative stress caused by cryopreservation.

• The Bulletin of The Korean Astronomical Society
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• v.37 no.2
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• pp.101.2-101.2
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• 2012

Shocks are ubiquitous in astrophysical plasmas: bow shocks are formed by the interaction of solar wind with planetary magnetic fields, and supernova explosions and jets produce shocks in interstellar and intergalactic spaces. The global morphologies of these shocks are usually described by a set of magnetohydrodynamic (MHD) equations which tacitly assumes local thermal equilibrium, and the resulting Rankine-Hugoniot shock jump conditions are applied to obtain the relationship between the upstream and downstream physical quantities. While thermal equilibrium can be achieved easily in collisional fluids, it is generally believed that collisions are infrequent in astrophysical settings. In fact, shock widths are much smaller than collisional mean free paths and a variety of kinetic phenomena are seen at the shock fronts according to in situ observations of planetary shocks. Hence, both the MHD and kinetic equations have been adopted in theoretical and numerical studies to describe different aspects of the physical phenomena associated with astrophysical shocks. In this paper, we present the results of 3D relativistic particle-in-cell (PIC) simulations for ion-electron plasmas, with focus on the shock structures: when a jet propagates into an unmagnetized ambient plasma, a shock forms in the nonlinear stage of the Weibel instability. As the shock shows the structures that resemble those predicted in MHD systems, we compare the results with those predicted in the MHD shocks. We also discuss the thermalization processes of the upstream flows based on the time evolutions of the phase space and the velocity distribution, as well as the wave spectra analyses.

• Journal of Embryo Transfer
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• v.28 no.4
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• pp.349-353
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• 2013

The objective of this study was to compare the effect of semen extenders on the sperm motility, viability, acrosome integrity and functional integrity of plasma membrane (HOST: hypo-osmotic swelling test) during liquid preservation of Korean Native boar semen. In this experiment, semen was diluted in Androhep plus, Beltsville Thawing Solution (BTS), ModenaTM, Seminark and Vitasem LD. Sperm-rich fractions were collected from three Korean Native boars and sub-samples were diluted ($30{\times}10^6$ spermatozoa/ml) in different semen extenders. Semen samples were stored at $17^{\circ}C$ for 96 hours. On everyday (0, 24, 48, 72, 96 h) after storage, the sperm characteristics relevant for fertility, such as sperm motility, viability, acrosome integrity and HOST positive were evaluated. The motility of spermatozoa stored in different extenders was no significantly different among other extenders (P>0.05). Also, no difference was observed among samples processed with different extenders in the percentage of sperm viability, acrosome integrity and HOST positive. All extenders maintained a high percentage (70%) of sperm motility, viability and acrosome integrity through 96 h of storage. The result of this study show that there was no significant differences among extenders in their capacity to preserve motility, viability and membrane integrity of spermatozoa from normal, fertile Korean Native boars for 96 h of liquid preservation at $17^{\circ}C$.

• Journal of Embryo Transfer
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• v.31 no.3
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• pp.169-177
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• 2016

There is a growing interest in the application of primary hepatocytes for treatment of liver diseases in humans and for drug development. Several studies have focused on long-term survival and di-differentiation blocking of primary hepatocytes in an in vitro culture system. Therefore, the present study also aimed to optimize an in vitro culture system using primary rat hepatocytes. Primary rat hepatocytes from 6-week-old male Crl:CD rats were isolated using a modified two-step collagenase perfusion. Healthy $3.5{\times}10^6$ primary rat hepatocytes were seeded into a 2 dimensional (2D) culture in a 25T culture flask coated with collagen type I or into a 3D culture in a 125-ml spinner flask for 7 days. Production of plasma protein (ALB and TF), apoptosis (BAX and BCL2), and CYP (CYP3A1) related genes were compared between the 2D and 3D culture systems. The 3D culture system had an advantage over the 2D system because of the relatively high expression of ALB and low expression of BAX in the 3D system. However, the level of CYP3A1 did not improve in the 3D culture with and without the presence of a dexamethasone inducer. Therefore, 3D culture has an advantage for albumin production and primary rat hepatocyte survivability, but a low expression of CYP3A1 indicated that primary rat hepatocytes require a high-density culture for stress reduction by continuous flow.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.249-255
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• 2014

Programmed cell death or apoptosis is associated with changes in $K^+$ concentration in many cell types. Recent studies have demonstrated that two-pore domain $K^+$ ($K_{2P}$) channels are involved in mouse embryonic development and apoptotic volume decrease of mammalian cells. In cerebellar granule neurons that normally undergo apoptosis during the early developmental stage, TASK-1 and TASK-3, members of $K_{2P}$ channels, were found to be critical for cell death. This study was performed to identify the role of $K^+$ channels in the $H_2O_2$-induced or cryo-induced cell death of mouse and bovine embryos. Mouse and bovine two-cell stage embryos (2-cells) exposed to $H_2O_2$ for 4 h suffered from apoptosis. The 2-cells showed positive TUNEL staining. Treatment with high concentration of KCl (25mM) inhibited $H_2O_2$-induced apoptosis of 2-cells by 19%. Cryo-induced death in bovine blastocysts showed positive TUNEL staining only in the cells near the plasma membrane. Cryoprotectant supplemented with 25 mM KCl reduced apoptosis slightly compared to cryoprotectant supplemented with 5 mM KCl. However, the combination of antioxidants (${\beta}$-mercaptoethanol) with 25 mM KCl significantly decreased the rate of $H_2O_2$-induced and cryo-induced apoptosis compared to treatments with only antioxidants or 25 mM KCl. These results show that blockage of $K^+$ channel efflux for a short-time reduces $H_2O_2$- and cryo-induced apoptosis in mouse and bovine embryos. Our findings suggest that apoptosis in mouse and bovine embryos might be controlled by modulation of $K^+$ channels which are highly expressed in a given cell type.