• Journal of Embryo Transfer
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• v.17 no.1
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• pp.45-53
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• 2002

The main goal of this study was to produce transgenic porcine embryos by direct injection of sperm-mediated exogenous DNA. Spermatozoa (6$\times$10$^{6}$ sperms of final concentration) were mixed with pcDNA LAC Z (20 ng/$\mu$l) and subjected into electroporation (300~750 volts, 25 $\mu$F, 0.4 cm electrode). After sperm injection, the oocytes were activated electrically (1.7 KV/cm, 30$\mu$sec, single pulse) in 0.3 M mannitol solution or not. The sperm injected eggs were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air fur 144 h. The rates of cleavage and development into blastocyst stage in activation group were significantly higher than those of non-activation group (79.6% and 24.1% vs. 46.3% and 14.4%, respectively, p<0.05). Control oocytes and shame injection were developed to blastocysts low (2.5%). Sixty five (27.1%) out of 240 embryos observed in activation and non-activation groups were showed positive by X-gal staining. However, all embryos in both groups were expressed partial or mosaic pattern. These results suggested that electrical stimulation far oocytes activation after sperm injection enhances the incidence of both fertilization and development fellowing sperm injection in the pig. Our study also suggested that sperm-mediated transfer of exogenous DNA by ICSI would be used as a valuable tool for the production of transgenic porcine embryos.

• Restorative Dentistry and Endodontics
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• v.24 no.1
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• pp.100-115
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• 1999

The purpose of this study was to investigate the distribution of calcitonin gene-related peptide containing nerve fibers in rat pulp after dentinl injury by means of immunohistochemistry and confocal laser scanning microscope. The Spague-Dawley rats weighing about 250-300gm were used. The animals were devided into normal control and experimental groups. Experimental animals were sacrified 1, 2, 4, 7, 10, 21days after dentinal injury (dentin cutting, and then acid etching with 35% phosphoric acid) on the maxillary molar teeth. The maxillary teeth and alveolar bone were removed and immersed in the 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4), then were decalcified with 15% formic acid for 10 days. Serial frozen $50{\mu}m$ thick sections were cut on a cryostat. The rabbit CGRP antibody was used as a primary antibody with a dilution of 1:2000 in 0.01M PB. The sections were incubated for 48 hours at $4^{\circ}C$, and placed into biotinylated antirabbit Ig G as a secondary anti body with dilution of 1:200 in 0.01M PB and incubated in ABC(avidin-biotin complex). The peroxidase reaction was visualized by incubating the sections in 0.05% 3,3 diaminobenzidine tetrahydrochloride containing 0.02% $H_2O_2$. For the confocal laser scanning microscopic examination, Primary antibody reaction was same as immunoperoxidase stainning, but fluorescein isothiocyanate(FITC)-conjugate antirabbit IgG as a secondary antibody was used. The confocal laser scanning microscope was used for the examination. A series of images of optical sections was collected with a 20x objective at $3{\mu}m$ intervals throughout the depth of specimen. FITC fluerescence was registrated through a 488nm and 568nm excitation filter, and images were saved on optical disk. The stereoscopic images and three dimentionnal images were reconstructed by computer software, and then were analyzed. The results were as follows : 1. In normal control group, CGRP containing nerve fibers were coursed through the root with very little branching, and then formed a dense network of terminals in coronal pulp. 2. A slight increase in CGRP containing nerve fibers at 1 and 2day postinjury was noted subjacent to the injury site. In the 4day group, there were an extensive increase in the number of reactive fibers, followed by a partial return toward normal levels at 7~10 day postinjury, and return by 21days. 3. The sprouting of the CGRP containing nerve fibers was evident within 2day after dentinal injury, and by 4days there was a maximal increased, but was decreased at 7days and returned to normal 10~21 day postinjury. 4. In confocal laser scanning microscopic exammination, the distinct distribution pattern and sprouting reaction of CGRP containing nerve fibers were observed in stereoscopic images and three dimentional images. These results suggest that CGRP containing nerve fiber can be important role in the response to dental injury and pain regulation.

• Korean Journal of Food Science and Technology
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• v.43 no.2
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• pp.161-168
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• 2011

The purpose of this study was to evaluate microbiological contamination of fresh vegetables in Korea. Twenty types of vegetables were tested for total aerobic bacteria, coliforms, Escherichia coli, yeast and mold, and pathogenic bacteria such as Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Salmonella, E. coli O157:H7, Cronobacter sakazakii, Shigella, and Campylobacter. Levels of total aerobic bacteria and coliform on 20 vegetables were between 3.74 and 8.04 log CFU/g, and 0.16 and 5.02 log CFU/g, respectively. The highest contamination levels of total aerobic bacteria were observed on water dropwort, sprouts, mungbean sprout, and ballflower root. There was no significant difference (p>0.05) in microbial contamination levels of total aerobic count, coliform, E. coli, yeast and mold between organic and nonorganic vegetables. When isolation methods using selective agars were applied, L. monocytogenes, B. cereus, Salmonella and Campylobacter were isolated from some fresh vegetable samples. Results of API kit tests showed that L. monocytogenes was identified on Chinese cabbage, cucumber, soybean sprouts, and iceberg lettuce while Salmonella was identified on Korean leek. Furthermore, Campylobacter jejuni was also identified in more than 50 of the 100 samples. However, when positive samples from API kit were tested for real-time PCR or 16S rRNA sequencing method, only B. cereus from perilla leaf, carrot, water dropwort, and sprouts showed positive results. These results indicate that selective agar and API kit detection methods might result in false positive results for some pathogens. Therefore, studies need to improve isolation or confirmation methods for such pathogens.

• Journal of Chest Surgery
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• v.29 no.8
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• pp.816-821
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• 1996

Lung transplantation is the established treatment for the end stage lung disedse find preservation of the organ is a major obstacle In performing lung transplantation. For solving this problem, we evaluated the histopathologic changes for various preservation solutions. Male mongrel dogs of similar size and weight (15∼20 kg) were used. The dog lungs were flushed with 4fl normal saline(group 1 'n:5): Modified Euro-Collins solution(group 2 n:5) and University of Wisconsin solution (group 3 : n=6), 60m11kg through a catheter placed in the main pulmonary artery aft r flushing of PGE 1 (20ng1kg). The lungs were preserved for 60 hours and measured dry and wet weights. Histologic specimens were taken every 6 hours and %toed for light microscopic evaluation. The edema ratio of the lungs peaked in 12 hours although there was no difference between the groups. Histologically, alveolar septal changes developed in one case (20%) after 1 hour preservation with normal saline. In case of the University of Wisconsin solution, the alveolar septal distortions and swellings were seen in 1 cases (20%) after 6 hours preservation compared with 3 cases (60%) after 6 hours preservation with Modified Euro-Collins solution. Changes of the pneumocytes were observed after 24 hours preser- vation in group 1, after 48 hours preservation in group 2 and after 60 hours preservation in group 3. We conclude that University of Wisconsin solution might have a superior preservation effect compare to normal saline and Modified Euro-Collins solutions.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.30 no.6
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• pp.465-473
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• 2004

Purpose : The essential triad for nerve regeneration is nerve conduit, supporting cell and neurotrophic factor. In order to improve the peripheral nerve regeneration, we used polyglycolic acid(PGA) tube and brain-derived neurotrophic factor(BDNF) gene transfected Schwann cells in sciatic nerve defects of SD rat. Materials and methods : Nerve conduits were made with PGA sheet and outer surface was coated with poly(lactic-co-glycolic acid) for mechanical strength and control the resorption rate. The diameter of conduit was 1.8mm and the length was 17mm Schwann cells were harvested from dorsal root ganglion(DRG) of SD rat aged 1 day. Schwann cells were cultured on the PGA sheet to test the biocompatibility adhesion of Schwann cell. Human BDNF gene was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into E1 deleted region of adenovirus shuttle vector, pAACCMVpARS. BDNF-adenovirus was multiplied in 293 cells and purified. The BDNF-Adenovirus was then infected to the cultured Schwann cells. Left sciatic nerve of SD rat (250g weighing) was exposed and 14mm defects were made. After bridging the defect with PGA conduit, culture medium(MEM), Schwann cells or BDNF-Adenovirus infected Schwann cells were injected into the lumen of conduit, respectively. 12 weeks after operation, gait analysis for sciatic function index, electrophysiology and histomorphometry was performed. Results : Cultured Schwann cells were well adhered to PGA sheet. Sciatic index of BDNF transfected group was $-53.66{\pm}13.43$ which was the best among three groups. The threshold of compound action potential was between 800 to $1000{\mu}A$ in experimental groups which is about 10 times higher than normal sciatic nerve. Conduction velocity and peak voltage of action potential of BDNF group was the highest among experimental groups. The myelin thickness and axonal density of BDNF group was significantly greater than the other groups. Conclusion : BDNF gene transfected Schwann cells could regenerate the sciatic nerve gap(14mm) of rat successfully.

• Korean Journal of Poultry Science
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• v.31 no.4
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• pp.283-291
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• 2004

Feeding trials were conducted with Euglena strains grown under different media. The effect of supplementation of Euglena on the laying performance, egg quality and fatty acid composition of egg yolk was studied. In experiment I, two hundred eighty 32-wk-old ISA Brown layers were randomly assigned to seven dietary treatments for 4 wks. Each treatment consisted of 4 replications with 10 birds each housed in two birds cages. Control diet was formulated to have $17\%$ CP and 2,750 kcal ME/kg. Euglena gracilis Z. (EG) was added to control diet at the level of 0.25, 0.5, $1.0\%$ and Euglena gracilis Z. bleached and DHA enriched (EGBD; a strain mutated by streptomycin and cultivated in DHA enriched medium) at the level of 0.5, 1.0, $2.0\%$ in the diet. In experiment 2, three hundred 84-wk-old ISA brown layers were randomly assigned to five dietary treatments: T1; Control, T2; T1 + EGBD $0.5\%$, T3; T1 + Euglena gracilis Z. DHA enriched (EGD; cultivated in DHA enriched medium) $0.5\%$, T4; T1 + EGD $1.0\%$, T5; T1 + EGD $2.0\%$. Each treatment had 5 replication of 12 birds each housed in two birds cages. In experiments 1 and 2, Euglena suppplementation did not significantly affect egg production but increased egg weight and feed intake. In experiment 1, EG was more effective in increasing egg yolk color score than EGBD. Egg yolk color of EG $1\%$ treatment showed the highest score. EGBD supplementation increased DHA concentration of egg yolk. EGBD $2\%$ treatment showed the highest DHA and the lowest palmitic and stearic acids concentration in the egg yolk. In experiment 2, EGBD $0.5\%$ treatment showed highest DHA level in egg yolk (P<0.05). It was conducted that EGBD is a single cell protein source rich in DHA, that can be used to produce DHA enriched eggs.

• Journal of the Korean Arthroscopy Society
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• v.9 no.2
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• pp.143-147
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• 2005

Purpose: In case of anterior cruciate ligament (ACL) reconstruction, graft tendon is generally fixed in tibial tunnel with knee extended. When reconstructing ACL using hamstring tendon, the authors aim to find out the effect of knee joint position during graft fixation on postoperative knee joint stability and range of motion. Materials and Methods: Prospective study was done on patients who have undergone ACL reconstruction using hamstring tendon from May 2002 to January 2003 We used Rigifix system (Mitek Product, Johnson and Johnson, USA) and Intrafix system for fixation. Thirty nine patients received ACL reconstruction during this period. Excluding 2 patients lost in the follow-up, 37 patients were analyzed. The mean follow-up period was 14 months $(13{\sim}25months)$. Knee position was decided alternatively without any bias. Clinical evaluation was based on Lachman test, pivot shift test, Lysholm score, IKDC(international knee documentation committee) assessment and side to side KT-1000 maximal manual arthrometer difference. Results: After the last follow-up, average postoperative Lysholm score was 93.1 poins(65-98points). According to IKDC score, 26 cases were normal, 10 cases were nearly normal, 1 case was abnormal and we had no case of severe abnormality. The mean difference from the normal side was 2.5 mm under maximal manual loading KT-1000 arthrometer. According to postoperative Lachman test, 32 cases were normal,2 cases were grade I and 1 case was grade II. There were 34 cases of normal, 2 cases of grade I and 1 case of grade II. When using maximal manual KT-1000 arthrometer side to side difference, the difference from the normal side while fixing the tibia at 20'knee flexion was 2.3 mm and at full extention the difference was 2.7 mm. The range of motion at postoperative 1 year showed 5 degree flexion contracture in 1 case at 20 degrees knee flexion and 10 degrees of flexion limitation was observed in 2 cases at full extension. Conclusion: When ACL reconstruction using autogenous hamstring tendon, anterior laxity showed no difference in its stability between two groups. Tibial side fixation at full extension may be helpful in preventing flexion contracture due to overconstrained graft tendon.

• Korean Journal of Ecology and Environment
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• v.39 no.1 s.115
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• pp.21-31
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• 2006

The objectives of this study were to analyze trends of temporal water quality and trophic state in Andong and Imha reservoirs using chemical dataset during 1993 ${\sim}$ 2004, obtained from the Ministry of Environment, Korea. According to long-term limnological analyses, Suspended solids (SS) in Imha Reservoir were 2 ${\sim}$ 8 fold2 greater, than those in SS of Andong Reservoir, and the high solids increased total phosphorus (TP) and biological oxygen demand ($BOD_5$) and decreased the transparency, measured as Secchi depth (SD). Chlorophyll-a (CHL-a) increased little or decreased slightly in the both reservoirs during the high solids, resulting in reduced yields of CHL-a : TP ratios. The deviation analysis of Trophic State Index (TSI) in Imha Reservoir showed that about 70% of TSI (CHL-a)-TSI (SD) and TSI (CHL-a)-TSI(TP) values were less than zero and the lowest values were-60, indicating that influence of inorganic solids (or non-volatile solids) on phytoplankton growth was evident in Imha Reservoir and the impact was greater than that of Andong Reservoir. Inorganic solids in Imha Reservoir resulted in light limitation on phytoplankton growth and thus contributed variations in the relations among three parameters of trophic state index. Especially, seasonal data analysis of nutrients in both reservoirs showed that during the postmonsoon, mean TP concentration was Imha Reservoir greater in than that in Andong Reservoir. The higher TP concentrantion was mainly attributed to increases of inorganic solids from soil erosions and nonpoint source inputs within the watershed. The high inorganic turbidity in Imha Reservoir should be reduced for the conservation of water quality for, especially a tap water supply.

• The Journal of the Petrological Society of Korea
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• v.9 no.2
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• pp.84-98
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• 2000

Daejeon-sa basalt in the Mt. Juwang area composed of 12 basalt flows alternate with 9 peperites and each basalt and peperite has the variety of thickness. Peperites yielded in Daejeon-sa basalt are mixed of basalt with reddish shale, of which textural type is globular peperite. Basalts yielded in Daejeon-sa basalt are massive basalt without vesicule, although sometimes vesicules are founded in upper within a flow unit. The basalt has mainly pseudomorph of olivine as phenocryst, and also plagioclase and clinopyroxene phenocryst. Matrix is mainly subophitic texture. The plotting result on the TAS diagram shows these basalts belong to the sub-alkaline, and it can be subdivided into calc-alkaline series on the basis of the diagram of Si02 vs. K20 and of alkali index vs. A1203 diagram. According to plots of wt.% oxides vs. wt.% MgO, abundances of A1203 and CaO increase with decreasing MgO while F ~ dOecre~ase . With decreasing MgO compatible elements decrease while incompatible elements increase. In spider diagram of MORB-normalized trace element patterns, HFS elements are nearly similiar with MORB, but LIL elements are enriched. Especially, contents of Ce, F: and Sm are enriched but Nb is depleted. In the chondrite-normalized REE patterns light REEs are enriched than heavy REEs. Tectomagmatic discrimination diagrams shows basalts in the study area are formed in the tectonomagmatic environment of subduction zone under continental margin. This result accord with characters of chemical composition mentioned above. Cr vs. Y diagram and CeM, vs. Ce diagram show that the primary magma of the basalts may formed by the about 15% partial melting of garnet-peridotite in the mantle wedge. After then, Daejeon-sa basalts may formed from evolved magma undergone mainly olivine fractional crystallization and contarnination of crustal materials before eruption.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.1
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• pp.23-30
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• 1995

This study was undertaken to clarify the effects of electrical stimulation on physicochemical and rheological properties of the plaice (Paralichthys olivaceus) muscle at early period after death. The plaices were electrically stimulated in seawater bath (110V/60Hz) for 15sec., 35sec., and 60sec. and killed instantly with spiking at the head. Killed samples were stored at $5^{\circ}C$, and the changes in rigor index, ATP breakdown, lactate accumulation, and breaking strength of muscle through storage were investigated. Electrical stimulation effectively accelerated rigor-mortis, lactate accumulation , and ATP breakdown. As the time of electrical stimulation was lengthened, the onset of rigor-mortis of all samples were accelerated Just after killing, and the amount of lactate was rapidly increased, But, significant differences were not observed in variance of rigor-mortis and lactate concentration. Electrically stimulated plaices showed decreasing in ATP to $4.58{\mu}mole/g$ for 15sec., $4.13{\mu}mole/g$ for 35sec., and $2.39{\mu}mole/g$ for 60sec. samples as compared with $5.5{\mu}mole/g$ of unstimulated samples. As the time of electrical stimulation was lengthened, ATP in samples were decomposed more rapidly. The rate constant of ATP breakdown were $0.244hr^{-1}$ for 15sec., $0.358hr^{-1}$ for 35sec., and $0.479hr^{-1}$ for 60sec.. The level of breaking strength in muscle of the plaice was $1050.30\pm50.23g$ immediately after killing. Values of breaking strength in samples electrically stimulated for 35sec. increased rapidly just after killing among all samples. However, the breaking strength was not increased through the whole storage time in samples stimulated for 60sec.. The value and time roaching to the maximum breaking strength for each samples stimulated electrically for 15, 35 and 60 second were $1264.43\pm35.76g$ and 2hr, $1357.68\pm22.50g$ and Ohr, and $1012.18\pm57.36g$ and Ohr. Breaking strength in all samples electrically stimulated decreased significantly (P<0.05) after reaching the maximum values.