• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.313-321
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• 1999

The purpose of these study was to investigate the use of a glass micropipette (GMP) as a vessel for vitrification of bovine IVP blastocysts, to compare the post-thaw survival rates of bovine blastocysts frozen in GMP with those frozen in OPS that have been previously investigated, and to improve the hatching rate following vitrification with GMP method. The GMP vessel permits higher freezing and warming rate than the OPS due to the higher heat conductivity of the glass and lower mass of the solution that contains the embryos. Groups of three bovine IVP blastocysts were sequentially placed into vitrification solution before being loaded into either the OPS or GMP vessels and immersed into L$N_2$within 20 to 25 sec. Post-thaw blastocysts were serially washed in 0.25 and 0.15 M sucrose in HM and TCM-199 for each 5 min, respectively, and then cultured in TCM 199 supplemented with 10% FCS for 24 h. The rate of blastocyst re-expanding did not significantly different for OPS (75.9%) and GMP (80.0%) methods (P＞0.05). The hatching rates in OPS (34.1%) and GMP (37.5%) methods were significantly lower than that in control group (54.3%) (P＞0.05). In addition, the rate of blastocyst re-expanding was significantly lower if blastocysts were vitrified in the wide portion of the micropipette rather than the narrow portion of the micropipette (83.3 vs 56.7%) (P＞0.05), even though three blastocysts were loaded per vessel. The hatching rate in 0.05% pronase solution treatment for 30, 60 and 90 see (45.9, 54.7 and 57.5%) were significantly higher than that in control (35.0%), even though there was not significantly different between 30 see and control. These results indicate that both vitrification vessels can provide high survival rates of bovine IVP blastocysts. However, the GMP vessel has the advantage over the OPS, in that the former does not need a cap to protect the vessel from floating after immersion in L$N_2$. The location of the embryos (narrow or wide portion of immersion) were considered to be limiting factors to the viability of bovine IVP embryos. The exposing in 0.05% pronase solution for 60 or 90 see can increase hatching rates of post-thaw bovine IVP blastocysts.

• Korean Journal of Animal Reproduction
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• v.20 no.2
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• pp.207-214
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• 1996

To examine the developmental capacity of manipulated embryos after ultrarapid refreezing and thawing, mouse embryos were biopsied at 4-cell stage, frozen twice at 4-cell and morula stages, respectively, and then transferred to rec-ipients. Single blastomeres were biopsied from 4-cell embryos by a modified aspiration method. Biopsied 4-cell embryos were equilibrated into freezing medium at room temperature for 2.5 min, loaded into 40 $\mu$I of freezing medium in 0.25 ml plastic straw and then directly immersed into liqiud nitrogen. Freezing medium for 4-cell embryos consisted of 4.0 M ethylene glycol and O.25 M sucrose in dPBS supplemented with 6 mg/lm BSA. Morulae were frozen into freezing medium containing 5.0 M glycerol instead of ethylene glycol. Thawing was conducted by agitating each straw in 3TC water for 20 sec. The c content of each straw was expelled into 0.5 ml of dilution medium, which consisted of 0.25 M sucrose and 3 mg/ml BSA in dPBS. The thawed embryos were rehydrated in dilution medium for 10 min, washed 3 times with dPBS and then cultured in M16 medium at 37$^{\circ}C$, 5% CO$_2$ in air. Blastocysts that developed from frozen or refrozne biopsied embryos were transferred to recipients on Day 3 of pseudopregnancy, respectively. In vitro and in vivo developmental rates of the biopsied and intact 4~cell embryos after freezing and thawing were 78 (10l/130) and 25% (10/40), and 91 (114/125) and 30% (12/40), respectively. Although the rates of in vitro development of biopsied and intact embryos to blastocyst stage were significantly different after freezing and thawing (P

• The Korean Journal of Pesticide Science
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• v.6 no.2
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• pp.96-104
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• 2002

This study was carried out to investigate cytotoxicity of paraquat on NIH 3T3 fibroblasts, toxicity of paraquat and compensatory effects of 3-methylcholanthrene (3-MC) on the rat lung. In order to conduct MIT [3-(4,5-Dimethylthiazol-2-yl) -2,5-diphenyl -2H-tetrazolium-bromide] and NR (Neutral red) assay, the $5.0{\times}10^4cell/ml$ of NIH 3T3 fibroblast in each well of 24 multi-dish were cultured. After 24 hours, the cells were treated with solution of paraquat (1, 25, 50 and $100{\mu}M$ respectively). After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours. MIT and NR assay were performed to evaluate the cytotoxicity of cell organelles. $MTT_{50}\;and\;NR_{50}$ of paraquat were $1668.97{\mu}M\;and\;1030.85{\mu}M$, respectively. These $IC_{50}$ of Paraquat were decided as a low cytotoxicity by Borenfreund and Puemer (1984). In order to observe the toxicity and compensatory effects of paraquat on the rat lung, Spraque Dawley male rats were used as experimental animals and were divided into paraquat only treated group and simultaneous application group of paraquat and 3-MC, at 30 min and 1, 3, 6, 12, 24, 48 and 96 hrs interval after each treatment. The animals were sacrificed by decapitation and their or the lungs were immediately removed, immersed in fixatives, and were processed with routine method for light microscopic study. Paraffin sections were stained with H&E and iron hematoxylin of Verhoeff. Under the light microscopy, erythrocytes were full in alveolar capillaries at 3 hrs and congested at 24 hrs after paraquat administration. The great alveolar cells (Type II cell) were increased and mitosis of great alveolar were observed in interalveolar septa. Many lymphocytes, macrophages and polymorphonuclear (PMN) cells were observed in connective tissue surrounding lung tissue and germinal center in lymph follicles of terminal bronchiole. Alveolar macrophages were increased in interalveolar septa and alveoli at 48 hrs. And observed many alveolar macrophages at 96 hrs. In iron hematoxylin stain of Verhoeff, Collagen fiber were increased in respiratory bronchiole, interalveolar septa and alveoli and breath of alveoli, and alveolar pore were broaden. But, in paraquat plus 3-MC treated group, morphological changes were mild in lung tissue. These results indicate that 3-MC has a compensatory effects against toxicity of paraquat by conjugation with oxygen.

• Journal of the korean academy of Pediatric Dentistry
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• v.32 no.1
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• pp.164-173
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• 2005

Mechanical preparation has been introduced to provide the sealant retention. The objective of this study was to compare the fissure penetration and the microleakage of pit and fissure sealant using mechanical preparation(mechanical preparation + acid etching) and acid etching only. An additional objective of this study was to compare the fissure penetration and the microleakage of unfilled and filled sealant in both methods. Sixty human premolars extracted for orthodontic purpose were selected. Thirty teeth were acid etched alone and remaining thirty teeth were prepared with a $\frac{1}{4}$ round bur and then acid etched. One-half of teeth in each surface treatment method were sealed with unfilled sealant and the other half were sealed with filled sealant. All of the teeth were thermocycled for 1200 cycles at $5^{\circ}C\;and\;55^{\circ}C$ and immersed in 5% methylene blue for 24 hours. Each tooth was sectioned bucco-lingually at mesial pit and distal pit and examined under a Measurescope. In the case of mechanical preparation, fissure penetration of sealant was significantly increased compared with the case of acid etching only(P < 0.05). The filled and unfilled sealant using mechanical preparation showed significantly decreased microleakage when compared with the unfilled sealant using acid etching only(P < 0.05). No differences were found in fissure penetration and microleakage between unfilled and filled sealant in both methods. Taken together, the results of this study suggest that mechanical preparation and filled sealant are recommended when placing pit and fissure sealant. However, further clinical studies should be performed in regard to microleakage.

• Journal of Chest Surgery
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• v.31 no.2
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• pp.108-112
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• 1998

Experimental trials of unilateral lung transplantation in dogs have been attempted and satisfactory results were obtained without any noticeable difficulty in surgical techniques. Fourteen dogs with the body weight of around 25 kg were anesthesized by 20~30 mg/kg of intravenous Entobar,; one was sacrificed to make available blood for use during transplantation for the recipient dog. A mid-sternotomy incision was performed and 20 mg/kg of Prostaglandin E1 was infused through the pulmonary artery and Euro-Collin's(E-C) preservation solution, cooled down to 4$^{\circ}C$, was perfused at the rate of 70cc/kg by a pressure of 30 cmH2O. The heart-lung block was then resected out and promptly immersed in the prepared preservation solution at 4$^{\circ}C$. One lung preserved in the EC solution at 4$^{\circ}C$ was anastomosed to the recipient dog in the order of the pulmonary vein, bronchus then pulmomary artery and the thoracotomy incision was closed after the bleeding control and tube thoracostomy. Then the pneumonectomy in the opposite side was perfomed in the same manner and the tailored lung was transplanted in the order of the pulmonary vein, bronchus, then pulmonary artery. We conclude that in the bilateral sequential lung transplantation, the right lung transplantation should precede to better expose the operative field and to prevent reperfusion injury; also, the cardiopulmonary bypass should be consider for certain appropriate cases.

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.191-198
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• 2001

The purpose of this study was to investigate the effects of embryo sources such as in vivo vs. in vitro produced blastocyst, and culture systems on the membrane permeability and viability of bovine blastocyst following GMP vitrification. To produce in vivo embryos, six cows were superovulated by administration of follicle stimulation hormone (FSH) and prostaglandin $F_{2{\alpha}}$(PG $F_{2{\alpha}}$). in vitro embryos were produced by two different culture systems, oviduct co-culture (OCS) and defined culture system (HECM-6; DCS). Ovaries were picked up at a local slaughterhouse and transported to laboratory in 3$0^{\circ}C$ saline within 2 h. Ovaries were washed with same saline three times and then placed in saline on warm plate adjusted at 3$0^{\circ}C$ during aspiration. The blastocysts produced were assigned for membrane permeability and viability following GMP vitrification. The membrane permeability of blastocysts was checked in 0.5 M sucrose solution on warm plate at 35$^{\circ}C$ for 0, 2, 5 and 7 min, respectively. Then the diameters (width and length) of embryo cytoplasms were measured by a eyepiece meter, and they were converted to their volume by 4/3 $\pi$ $r^3$. The blastocysts were cryopreserved by GMP vitrification method, where they were sequentially placed into vitrification solution before being loaded into GMP vessels and immersed into L$N_2$ within 20 to 25 sec. Post-thaw blastocysts were serially washed in 0.25 and 0.15 M sucrose in HM and TCM-199 for 5 min each, and then cultured in TCM 199 supplemented with 10% FCS for 24 or 48 h. The volume change of in vivo blastocyst at 0, 2, 5 and 7 min (100, 37.1, 34.3 and 31.6%) was significantly more shrunk than those of in vitro blastocysts derived from OCS (100, 59.8, 48.9, 47.9%) and DCS (100, 57.2, 47.3 and 46.9%) (P＜0.05). The viability of post-thaw blastocyst derived from in vivo (93.6%) was also significantly different from those in OCS and DCS (81.9 and 83.6%; P＜0.05). In the present culture system, the morphology of embryos produced in vitro was similar to that of in vivo embryos, but the quality in membrane permeability and post-thaw viability showed a big difference from their sources as in vivo or in vitro derived from OCS and DCS. The results indicated that the quality of in vivo embryos in membrane permeability and post-thaw viability was better than those of in vitro embryos derived from OCS or DCS.

• Korean Journal of Environmental Agriculture
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• v.20 no.3
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• pp.155-161
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• 2001

This study was carried out to investigate cytotoxicity of paraquat or bentazone on NIH 3T3 fibroblasts, toxicity of paraquat or bentazone, and compensatory effects of 3-Methylcholanthrene(3-MC) on the rat liver. In order to MTT assay, the $5.0{\times}10^4$ cell/mL of NIH 3T3 fibroblast in each well of 24 multidish were cultured. After 24 hours, the cells were treated with solution of paraquat or bentazone(1, 25, 50, 100 ${\mu}M$ respectively). After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours. MTT assay were performed to evaluate the cytotoxicity of cell organelles. Paraquat or bentazone $MTT_{50}$ were 1668.97 ${\mu}M$ and 1506.97 ${\mu}M$, respectively. These $IC_{50}$ of paraquat or bentazone were decided low cytotoxicity by Borenfreund. In order to observe the toxicity and compensatory effects of paraquat or bentazone on the rat liver, Sprague-Dawley male rats were used as experimental animals and divided into paraquat or bentazone only treated group and simultaneous application group of paraquat or bentazone and 3-MC. At 30 min and 1, 3, 6, 12, 24, 48 and 96 hrs interval after each treatment, the animals were sacrificed by decapitation and liver were immediately removed, immersed in fixatives, and processed with routine method for light microscopic study. Paraffin sections were stained with H-E, PAM and Best Carmine. Under the light microscope, degenerative changes of hepatic lobules were frequently observed in portal area from 3 hrs after paraquat or bentazone treatment. All hepatic cells were induced degenerative change at 12 hrs and more severe degenerative change at 48 hrs after paraquat or bentazone treatment. Especially, hepatic cells of bentazone only treated group were distinctly showed pyknotic. Glycogen granules were increased in portal area at 3 hrs, all hepatic cells at 12 hrs and remarkably increased at 48 hrs after paraquat or bentazone treated group. But hepatic cells of bentazone only treated group were regeneration at 48 hrs from portal area and glycogen granules of hepatic cells of paraquat or bentazone and 3-MC combination treated group showed in central area only at 48 hrs. The results indicate that 3-MC may be decrease paraquat or bentazone cytotoxicity on the rat liver.

• Restorative Dentistry and Endodontics
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• v.26 no.4
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• pp.263-272
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• 2001

The aim of this study was to investigate the influence of four different light curing modes on the marginal leakage of Class V composite resin restoration. Eighty extracted human premolars were used. Wedge-shaped class Y cavities were prepared on the buccal surface of the tooth with high-speed diamond bur without bevel. The cavities were positioned half of the cavity above and half beyond the cemento-enamel junction. The depth, height, and width of the cavity were 2 mm, 3 mm and 2 mm respectively. The specimens were divided into 4 groups of 20 teeth each. All the specimen cavities were treated with Prime & Bond$^{R}$ NT dental adhesive system (Dentsply DeTrey GmbH, Germany) according to the manufacturer's instructions and cured for 10 seconds except group VI which were cured for 3 seconds. All the cavities were restored with resin composite Spectrum$^{TM}$ TPH A2 (Dentsply DeTrey GmbH, Germany) in a bulk. Resin composites were light-cured under 4 different modes. A regular intensity group (600 mW/${cm}^2$, group I) was irradiated for 30 s, a low intensity group (300 mW/${cm}^2$, group II) for 60 s and a ultra-high intensity group (1930 mW/${cm}^2$, group IV) for 3 s. A pulse-delay group (group III) was irradiated with 400 mW/${cm}^2$ for 2 s followed by 800 mW/${cm}^2$ for 10 s after 5 minutes delay. The Spectrum$^{TM}$ 800 (Dentsply DeTrey GmbH, Germany) light-curing units were used for groups I, II and III and Apollo 95E (DMD, U.S.A.) was used for group IV. The composite resin specimens were finished and polished immediately after light curing except group III which were finished and polished during delaying time. Specimens were stored in a physiologic saline solution at 37$^{\circ}C$ for 24 hours. After thermocycling (500$\times$, 5-55$^{\circ}C$), all teeth were covered with nail varnish up to 0.5 mm from the margins of the restorations, immersed in 37$^{\circ}C$, 2% methylene blue solution for 24 hours, and rinsed with tap water for 24 hours. After embedding in clear resin, the specimens were sectioned with a water-cooled diamond saw (Isomet$^{TM}$, Buehler Co., Lake Bluff, IL, U.S.A.) along the longitudinal axis of the tooth so as to pass the center of the restorations. The cut surfaces were examined under a stereomicroscope (SZ-PT Olympus, Japan) at ${\times}$25 magnification, and the images were captured with a CCD camera (GP-KR222, Panasonic, Japan) and stored in a computer with Studio Grabber program. Dye penetration depth at the restoration/dentin and the restoration/enamel interfaces was measured as a rate of the entire depth of the restoration using a software (Scion image, Scion Corp., U.S.A.) The data were analysed statistically using One-way ANOVA and Tukey's method. The results were as follows : 1. Pulse-Delay group did not show any significant difference in dye penetration rate from other groups at enamel and dentin margins (p>0.05) 2. At dentin margin, ultra-high intensity group showed significantly higher dye penetration rate than both regular intensity group and low intensity group (p<0.05). 3. At enamel margin, there were no statistically significant difference among four groups (p>0.05). 4. Dentin margin showed significantly higher dye penetration rate than enamel margin in all groups (p<0.05).

• The Journal of Korean Academy of Prosthodontics
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• v.47 no.2
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• pp.182-190
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• 2009

Statement of problem: Microleakage at the occlusal and gingival margin of Class V cavities restored with composite resin has traditionally been considered an obstacle to successful restoration. Purpose: The aim of this study was to assess the effectiveness of three different surface sealants(Fortify, Permaseal and Biscover LV) on the marginal sealing of Class V light-activated composite resin restorations(Z250). Material and methods: Forty noncarious human premolars and molars extracted within a three-month period were selected. Class V cavities with the occlusal margin in enamel and gingival margin in cementum were prepared in both buccal and lingual surfaces. The teeth, randomly assigned in four groups with twenty cavities in each group, were restored with composite resin after applying an adhesive system(Clearfil SE bond). After the finishing and polishing procedures, the restorations were covered with a specific surface sealants, except for the control samples, which were not sealed. After placing restorations, the specimens were thermocycled, and immersed in a 2% methylene blue solution for twenty four hours and sectioned longitudinally. The marginal microleakage was evaluated at the occlusal and gingival interfaces using a microscope and compared among the four groups using ANOVA test and Wilcoxon Rank-Sum test($\alpha$=0.05). Results: Statistical analysis showed that there was significantly less leakage when the surface sealants were used than there was in control group(P<.05). There were no significant differences of microleakage at occlusal and gingival margins among groups. There were no significant differences between microleakage of occlusal and gingival margins in each group. Fortify was not statistically different from control group at the gingival margin(P>.05). Conclusion: Application of surface sealants was an effective method of surface coating in reducing microleakage at occlusal and gingival margins of Class V composite resin restorations. However, it is certain that some microleakage still occurred despite the application of surface sealants, especially gingival margins.

• The Journal of the Convergence on Culture Technology
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• v.8 no.2
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• pp.227-234
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• 2022

Seok Jeon Hwang Wook (18913~1999), a descendant of a traditional literary writer in the western part of Honam, did not join the flow of modern and contemporary calligraphy and painting. And throughout his life, he enjoyed himself without losing the appearance of a scholar, immersed himself in traditional calligraphy, and gained spotlight at his late age for his original hand grabbing calligraphy. Immediately after the Korean War, all of his property was lost due to his two sons' left-wing activities, causing great pain at home. Even in the most painful and difficult time in human history, he relied on brushes, poetry, and gayageum to keep his upright scholarly spirit and national love. And beyond the pleasures of the worldly senses, he played with self-satisfaction in the 'true pleasure(大樂)' without greed. In the course of his studies, he focused on honing the fonts of Wang Hui-ji, Gu Yang-sun, An Jin-gyeong, Jo Maeng-bu, and Xin-wi and Lee Sam-man without a special teacher. In particular, he faced a crisis of having to give up his brush due to tremor that came after his 60th birthday, but he showed a strong will. He transformed it into a new style of art, such as developing hand grabbing calligraphy(握筆法) with a strong and strong energy that no one could match. From 1965 to 1983, 'right hand grabbing calligraphy' was used, and from 1984 to 1993, 'left hand grabbing calligraphy' was used. She made her name as a calligrapher widely known in 1973 (age 76) with her first solo exhibition, The Calligraphy Exhibition commemorating her 60th wedding anniversary. His writing method is naturally rough and sloppy by breaking away from the previous calligraphy methods and artificial technique, and is unfamiliar yet full of muscle. And the calm, strong and rough chuhoegsa(錐劃沙) and the heavy yet majestic ininni(印印泥) individual handwriting expressed a strange feeling and achieved original Seokjeon calligraphy that went beyond the existing calligraphy writing methods, and his indomitable calligraphy spirit was As a unique existence in the history of calligraphy, he still remains as a model.