• Journal of Embryo Transfer
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• v.30 no.3
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• pp.121-128
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• 2015

The objective of this study was to evaluate the efficiency of sperm cryosurvival in boar sperm separated by Percoll containing antioxidant enzymes. The boar semen was collected into a pre-warmed ($37^{\circ}C$) thermos bottle by gloved-hand method and was separated by 65% Percoll with superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) before freezing. The frozen sperm was thawed at $38.5^{\circ}C$ for 45 sec in water-bath for sperm characteristic analysis. The sperm were estimated with SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction, Rhodamine123/PI double staining for mitochondrial integrity and were analyzed using flow cytometry. In results, sperm viability, acrosome reaction and mitochondrial integrity were improved in separated sperm groups compared with unseparated sperm by Percoll (UP) group. Especially, viability was significantly higher in sperm separated by Percoll containing 400 IU CAT group compared with other groups (P<0.05). And acrosome reaction was decreased in sperm separated by Percoll with 300 IU SOD, 400 IU CAT and 0.5 mM GSH groups compared with other groups, however, there were no significantly difference mitochondrial integrity among sperm separated by Percoll with antioxidant enzymes. In conclusion, we suggest that use of Percoll containing antioxidant enzymes for sperm separation will be beneficial for sperm cryopreservation in pigs.

• Journal of Embryo Transfer
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• v.29 no.4
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• pp.385-391
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• 2014

Objective of this study was to investigate the effect of nicotinic acid (NA) on the characteristics in fresh semen of miniature pig. We evaluated viability, acrosome reaction and mitochondrial integrity of sperm on 0, 3, 7 and 10 days during storage period with nicotinic acid. As results, the survival rate of sperm in 15 mM NA (day 3, $87.8{\pm}1.2%$; day 5, $84.0{\pm}2.7%$; day 7, $82.2{\pm}0.9%$) and 30 mM NA (day 3, $87.7{\pm}0.3%$; day 5, $84.4{\pm}2.5%$; day 7, $82.3{\pm}0.7%$) groups were higher than control and 5 mM NA groups in 3, 7 and 10 days of semen storage. The NA-treated sperm on 10 day was used day for observing acrosome integrity. The survival sperm with acrosome reaction was higher in 30 mM NA group (day 3, $2.7{\pm}0.2%$; day 5, $3.3{\pm}0.6%$; day 7, $11.4{\pm}0.3%$) than in the control, significantly (P<0.05). Moreover, the live sperm with mitochondrial integrity was higher in whole treatment groups of NA than control group, significantly (P<0.05). Specially, most mitochondrial integrity on 10 day of semen storage was significantly higher in 30 mM NA group ($90.2{\pm}1.6%$) than other treatment groups (control, $81.8{\pm}3.1%$; 5 mM NA, $83.4{\pm}3.0%$; 15 mM NA, $89.1{\pm}0.7%$, P<0.05). In conclusion, supplement of NA in liquid semen of miniature pig can improve and maintain semen quality, such as viability, acrosome reaction, and mitochondria integrity.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.73-79
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• 2017

Transplantation is considered to be a very useful approach to improve human welfare and to prolong life-span. Heterologous organ transplantation using pig organs which are similar to human beings and easy to make mass-production has known as one of the alternatives. To ensure potential usage of the pig organ for transplantation application, it is essentially required to generate transgenic pig modifying immuno-related genes. Previously, we reported production of heterozygous ${\alpha}1,3$-galactosyltransferase (GalT) knock-out and human membrane cofactor protein (MCP) expressing pig ($GalT^{-MCP/+}$), which is enforced for suppression of hyperacute and acute immunological rejection. In this study, we reported generation of homozygous pig ($GalT^{-MCP/-MCP}$) by crossbreeding $GalT^{-MCP/+}$ pigs. Two female founders gave birth to six of $GalT^{-MCP/-MCP}$, and seven $GalT^{-MCP/+}$ pigs. We performed quantitative real-time PCR, western blot, and flow cytometry analyses to confirm GalT and MCP expression. We showed that fibroblasts of the $GalT^{-MCP/-MCP}$ pig do not express GalT and its product Gal antigen, while efficiently express MCP. We also showed no expression of GalT, otherwise expression of MCP at heart, kidney, liver and pancreas of transgenic pig. Taken together, we suggest that the $GalT^{-MCP/-MCP}$ pig is a useful candidate to apply xenotransplantation study.

• Applied Chemistry for Engineering
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• v.19 no.3
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• pp.270-276
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• 2008

$N_2$ doped $TiO_2$ nano-sized powder was prepared using a DC arc plasma jet and investigated with XRD, BET, SEM, TEM, and photo-catalytic decomposition. Recently the research interest about the nano-sized $TiO_2$ powder has been increased to improve its photo-catalytic activity for the removal of environmental pollutants. Nitrogen gas, reacting gas, and titanium tetrachloride ($TiCl_4$) were used as the raw materials and injected into the plasma reactor to synthesize the $N_2$ doped $TiO_2$ power. The particle size and XRD peaks of the synthesized powder were analyzed as a function of the flow rate of the nitrogen gas. Also, the characteristics of the photo-catalytic decomposition using the prepared powder were studied. For comparing the photo-catalytic decomposition performance of $TiO_2$ powder with that of $TiO_2$ coating, $TiO_2$ thin films were prepared by the spin coating and the pulsed laser deposition. For the results of the acetaldehyde decomposition, the photo-catalytic activity of $TiO_{2-x}N_x$ powder was higher than that of the pure $TiO_2$ powder in the visible light region. For the methylene blue decomposition, the decomposition efficiency of $TiO_2$ powder was also higher than that of $TiO_2$ film.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.87-94
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• 2017

Tissue engineering (TE) has been developed to create functional organs and tissue by combining 3D matrix and cells in vitro. Vascularization and angiogenesis are utmost important for supply of nutrients and oxygen in tissue engineered organs. The present study was performed to isolate and characterize primary endothelial cells (EC) from aorta of alpha 1, 3-enzyme galactosyltransferase knock out (GalT KO) pig, to minimize immune rejection and analyze body immune system for future xenotransplantation studies. Isolation of primary EC from aorta were performed by incubation with dispase for 8-10 min at $37^{\circ}C$. Primary EC were cultured in EC growth medium on different extra cellular matrix (ECM), either collagen or gelation. Primary EC exhibits morphological characteristics and showed positive expressions of EC specific marker proteins i.e. PECAM1, KDR and VWF despite of their ECM surface; however, on collagen based surface they showed increase in mRNA level analyzed by qPCR. Primary EC cultured on collagen were sorted by flow cytometer using KDR marker and cultured as KDR positive cells and KDR negative cells, respectively. KDR positive cells showed dramatically increased in PECAM1 and VWF level as compared to KDR negative cells. Based on the above results, primary EC derived from GalT KO are successfully isolated and survived continuously in culture without becoming overgrown by fibroblast. Therefore, they can be utilize for xeno organ transfer, tissue engineering, and immune rejection study in future.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.18 no.4
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• pp.118-124
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• 2017

In this study, a numerical simulation was performed using commercial code Fluent(v.17.1). The underground Combined Cycle Power Plant (CCPP) was simplified to analyze the methane gas leakage with the crack size and position. In addition, extensive numerical simulations were carried out for different crack sizes from 10 mm to 20 mm. The crack position is the gas leakage, which is assumed to be near the pipe elbow and the gas turbine. A total of 4 cases were compared and analyzed. To analyze the gas leakage, the concept of the Lower Flammable Limit (LFL) was applied. The leakage distance was defined in the longitudinal direction, and the transverse direction was estimated and quantitatively analyzed. As a result, the leakage distance in the longitudinal direction varies by 52.3 % depending on the crack size at the same crack position. Moreover, the maximum difference was 34.8 % according to the crack position when the crack sizes are identical. As jet flow impacts on the obstacle and changes its direction, the recirculation flows are formed. These results are expected to provide useful data to optimize the location and number of gas detections in confined spaces, such as underground CCPP.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.6
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• pp.643-651
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• 2001

Paddlewheel-driven circulation in a culture pond has been simulated based on the depth integrated 2 dimensional hydrodynamic model. Acceleration by paddlewheel is expressed as shaft force divided by water mass discharged by paddlewheel blades. The model has been calibrated and applied to culture ponds as following steps:- i) The model predicted velocities at every 10 m along longitudinal direction from the paddlewheel. The model was calibrated comparing the results with the measured values at mass correction factor $\alpha$ and dimensionless eddy viscosity constant $\gamma$, respectively, in a range $15\~20$ and 6. ii) Wind shear stress was simulated under conditions of direction $0^{\circ}C,\;90^{\circ}C\;and\;180^{\circ}C$ and speed 0.0, 2.5, 5.0 and 7.5 m/s. Change rate of current speed was <$1\%$ at wind in parallel or opposite direction to the paddlewheel-driven jet flow, while $4\%$ at orthogonal angle. iii) The model was then applied to 2 culture ponds located at the Western coast of Korea. The measured and predicted currents for the ponds were compared using the regression analysis. Analysis of flow direction and speed showed correlation coefficients 0.8928 and 0.6782 in pond A, 0.8539 and 0.7071 in pond B, respectively. Hence, the model is concluded to accurately predict circulation driven by paddlewheel such that it can be a useful tool to provide pond management strategy relating to paddlewheel operation and water quality.

• Journal of Embryo Transfer
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• v.30 no.2
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• pp.73-82
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• 2015

This study was designed to evaluate the possibility of increase through dairy female offspring's ratio by transfer of pre-selected transferrable blastocyst that was produced by pre-selected X-bearing semen with OPU derived oocytes. Elite dairy female cow is demanded strongly compared with male, the so called, farmer wants to produce only an elite female dairy offspring as a candidate female dairy cow for producing milk. In our study, we selected 2 elite dairy bull semen from National Agricultural Cooperative Federation to pre-select X-bearing semen and 5 elite dairy female cows as donor for collecting of OPU derived oocytes. OPU derived embryo production system was carried out an aspiration of immature oocytes from 5 donor cows 2 times per week, total 200 times for 2 to 7 months by an ultrasonographic guided follicular aspiration system and then produced in vitro-produced blastocysts by in vitro maturation, fertilization and culture. Dairy donor semen selected H-319, 320 bull in National Agricultural Cooperative federation was sorted X-bearing semen by flow-cytometer and frozen for using IVF with OPU derived oocytes. Donor cows were selected 5 elite dairy cows from Gyeongju Dairy Cow Community and then disease tests such as 4 kinds of disease before selecting was checked. Oocyte proportion of grade 1 to 3 from total collected oocytes was significantly lower in donor A and B than those in donor C, D and E (82.16 and 70.03% vs. 90.0, 91.78 and 93.57%), respectively (p<0.05). However, number of oocytes per session in donor A, C and E was significantly higher than those in donor B and D ($7.77{\pm}3.26$, $5.85{\pm}2.10$ and $7.03{\pm}2.14$ vs. $4.68{\pm}2.61$ and $5.21{\pm}1.97$ oocytes), but donor A was significantly higher than donor C (p<0.05). Development to blastocyst in donor B, C and E was significantly higher than those in donor A and D (31.0, 25.0 and 25.0% vs. 14.3 and 4.5%), but donor A was not different in donor C and E (p<0.05). Nine out of 10 blastocysts (90.0%) derived from OPU blastocysts were confirmed male embryos that was induced with Y-bearing semen to confirm sex ratio only. Total 96 blastocysts derived from female bearing semen were transferred into synchronized recipients and then confirmed 42 recipients (43.8%) pregnancy rate, 36 offspring (37.5%) and 91.7% female sex ratio (33 female vs. 3 male offspring). Taken together all data, elite dairy female offspring could be produced effectively by in vitro production system between pre-selected x-bearing semen and OPU derived oocytes that would be influential breeder in the breeding of dairy farm to increase effectively elite dairy offspring ratio as well as net income in the dairy farmer.

• Magazine of the Korean Society of Agricultural Engineers
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• v.24 no.2
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• pp.49-55
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• 1982

The purpose of this study is to classify the factors influenced on the damages of head works suffered from the storm flood occurred on July 22 1980 in both Musim and Bochong rivers and to find out an integral counter measures against the causes influenced on the disaster of head works in the engineering aspect of planning, design, construction and maintenance. In this survey, number of samples was taken 25 head Works, and the counter measures against the causes of their disasters summarized was as follows, 1. In the aspect of planning a. As the flood water level after the establishment of head works is more increased than the level before setting of head works owing to having more gentle slope of river bed between the head works than nature slope of river bed. Number of head works should be reduced for the appropriate annexation of them b. In the place where head works is established on the curved point of levee, the destruction of levee becomes severe by the strong deflective current. Therefore the setting of head works on the curved point should be kept off as long as possible and in case of unavoidable circumstances the construction method such as reinforced concrete wall or stone wall filed with concrete and anchored bank revetments should be considered. 2. In the aspect of design a. As scoring phenomena at up stream is serious around the weir Where the concentration of strong current is present in such a place, up stream apron having impermeability should be designed to resist and prevent scoring. b. As the length of apron and protected bed is too short to prevent scoring as down stream bed, the design length should be taken somewhat more than the calculated value, but in the case the calculated length becomes too long to be profitable, a device of water cushion should be considered. c. The structure of protected river bed should be improved to make stone mesh bags fixed to apron and to have vinyl mattress laid on river bed together with the improvement for increasing the stability of stone mesh bags and preventing the sucked sand from the river bed. d. As the shortage of cut-off length, especialy in case of the cutoffs conneting both shore sides of river makes the cause of destruction of embankment and weir body, the culculation of cut-off length should be taken enough length based on seepage length. 3. In the aspect of design and constructions a. The overturing destruction of weir by piping action was based on the jet water through cracks at the construction and expansion joints. therefore the expansion joint should be designed and constructed with the insertion of water proof plate and asphalt filling, and the construction joint, with concaved shape structure and steel reinforcement. b. As the wrong design and construction of the weep holes on apron will cause water piping and weir destruction, the design and construction of filter based on the rule of filter should be kept for weep holes. c. The wrong design and construction of bank revetment caused the severe destruction of levee and weir body resulting from scoring and impulse by strong current and formation of water route behind the revetment. Therefore bank revetment should be designod and constructed with stone wall filled with concrete and anchored, or reinforced concrete wall to prevent the formation of water flow route behind the wall and to resist against the scoring and impulse of strong stream. 4. In the aspect of maintenance When the damaged parts occurred at head works the authorities and farmers concerned should find and mend them as soon as possible with mutual cooperation, and on the other hand public citizen should be guided for good use of public property.

• Journal of Embryo Transfer
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• v.30 no.1
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• pp.7-15
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• 2015

The objective of this study was to investigate the efficiency of nicotinic acid on sperm cryosurvival and fertilization ability in frozen-thawed boar semen. Boar semen was collected by glove-hand method and was frozen using freezing solution treated to 0, 5, 10 and 20 mM of nicotinic acid. The frozen sperm for sperm characteristic analysis was thawed such as viability, acrosome reaction, and mitochondrial integrity. The frozen-thawed sperm was estimated by SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction and Rhodamine123/PI double staining for mitochondrial integrity using a flow cytometry. The embryo was estimated in vitro development and DCFDA staining for reactive oxygen species assessment. As results, frozen-thawed sperm viability was significantly higher in 5 and 10 mM ($61.1{\pm}1.5%$,$64.7{\pm}2.0%$) of nicotinic acid than other groups (0 mM, $52.1{\pm}2.3%$; 20 mM, $47.8{\pm}5.1%$, P<0.05). The live sperm with acrosome reaction was significantly higher in 5 and 10 mM of nicotinic acid ($26.1{\pm}1.8%$, $24.9{\pm}1.5%$) than other groups (0 mM, $35.3{\pm}0.8%$; 20 mM, $36.5{\pm}1.9%$, P<0.05). The live sperm with mitochondrial integrity was significantly higher in 5 and 10 mM ($84.2{\pm}3.6%$, $88.4{\pm}2.3%$) of nicotinic acid than other groups (0 mM, $77.3{\pm}4.4%$; 20 mM, $73.3{\pm}3.6%$, P<0.05). Blastocyst rate of in vitro development was significantly higher in 10 mM ($17.0{\pm}1.3%$) of nicotinic acid than other groups (0 mM, $9.4{\pm}0.5%$; 5mM, $12.6{\pm}0.8%$; 20 mM, $5.0{\pm}1.0%$, P<0.05). Moreover, total cell number was higher in 5 and 10 mM ($53.6{\pm}2.9%$, $57.9{\pm}2.8%$) of nicotinic acid than other groups (0 mM, $41.0{\pm}1.4%$; 20 mM, $23.2{\pm}2.8%$, P<0.05). Hydrogen peroxide in embryos was lower in 5 mM nicotinic acid ($0.7{\pm}0.1%$) than other groups (0 mM, $1.0{\pm}0.1%$; 10mM, $0.9{\pm}0.0%$; 20 mM, $1.4{\pm}1.0%$, P<0.05). In conclusion, nicotinic acid-treated semen improves cryosurvival and quality of spermatozoa. Also, the fertilized oocytes with nicotinic acid improve quality of embryo and blastocyst formation.