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    Effect of dietary substitution of sea tangle (ST), Laminaria japonica with rice bran (RB) on growth and body composition of juvenile abalone, Haliotis discus

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    ๋ณธ ์—ฐ๊ตฌ์—์„œ๋Š” ์ „๋ณต์šฉ ๋ฐฐํ•ฉ์‚ฌ๋ฃŒ๋‚ด ๋‹ค์‹œ๋งˆ๋Œ€์ฒด์›์œผ๋กœ์„œ ์ƒ๋ฏธ๊ฐ•์„ ์ด์šฉํ•˜์—ฌ ์ „๋ณต์˜ ์„ฑ์žฅ ๋ฐ ์ฒด์กฐ์„ฑ์— ๋ฏธ์น˜๋Š” ์˜ํ–ฅ์„ ์กฐ์‚ฌํ•˜์˜€๋‹ค. ์‹คํ—˜์— ์ด์šฉ๋œ ์ „๋ณต ์น˜ํŒจ๋Š” ์‹คํ—˜์กฐ๊ฑด์—์„œ 4์ฃผ๊ฐ„ ์ ์‘์‹œํ‚จ ํ›„, ๋งˆ๋ฆฌ๋‹น ํ‰๊ท  0.43 g์˜ ์ „๋ณต 1,260๋งˆ๋ฆฌ๋ฅผ ๋ฌด์ž‘์œ„๋กœ ์„ ๋ณ„ํ•˜์—ฌ 18๊ฐœ์˜ 70 L ํ”Œ๋ผ์Šคํ‹ฑ ์ˆ˜์šฉ๊ธฐ์— ๊ฐ๊ฐ 70๋งˆ๋ฆฌ์”ฉ ์ˆ˜์šฉํ•˜์˜€๋‹ค. ์‹คํ—˜์‚ฌ๋ฃŒ๋Š” 1์ผ 1ํšŒ ๋งŒ๋ณต ์ˆ˜์ค€์œผ๋กœ ์‚ฌ๋ฃŒ๋ฅผ ๊ณต๊ธ‰ํ•˜์˜€๋‹ค. ์ด ์‚ฌ์œก์‹คํ—˜ ๊ธฐ๊ฐ„์€ 16์ฃผ๊ฐ„์ด์—ˆ๋‹ค. ์ด 6์ข…๋ฅ˜์˜ ์‹คํ—˜์‚ฌ๋ฃŒ๋ฅผ ์ค€๋น„ํ•˜์˜€๋‹ค. ๋‹ค์‹œ๋งˆ ๋ถ„๋ง์„ 20% ์ฒจ๊ฐ€ํ•˜์—ฌ ์ œ์กฐํ•œ ๋Œ€์กฐ๊ตฌ ์‚ฌ๋ฃŒ(RB0), ๋‹ค์‹œ๋งˆ๋ฅผ ์ƒ๋ฏธ๊ฐ•์œผ๋กœ ๋Œ€์ฒดํ•œ ๋‹ค์‹œ๋งˆ๋Œ€์ฒด 20% (RB20), ๋‹ค์‹œ๋งˆ๋Œ€์ฒด 40% (RB40), ๋‹ค์‹œ๋งˆ๋Œ€์ฒด 60% (RB60), ๋‹ค์‹œ๋งˆ๋Œ€์ฒด 80% (RB80) ๋ฐ ๋‹ค์‹œ๋งˆ๋Œ€์ฒด 100% (RB100) ์‚ฌ๋ฃŒ๋ฅผ ์ค€๋น„ํ•˜์˜€๋‹ค. ์ „๋ณต์˜ ์ƒ์กด๋ฅ ์€ ๋ชจ๋“  ์‹คํ—˜์šฉ ๋ฐฐํ•ฉ์‚ฌ๋ฃŒ๋ฅผ ๊ณต๊ธ‰ํ•œ ์‹คํ—˜๊ตฌ๊ฐ„์˜ ์œ ์˜์ ์ธ ์ฐจ์ด๊ฐ€ ๋‚˜ํƒ€๋‚˜์ง€ ์•Š์•˜์œผ๋ฉฐ, ์ฆ์ฒด๋Ÿ‰(weight gain)์€ RB40์„ ๊ณต๊ธ‰ํ•œ ์‹คํ—˜๊ตฌ๊ฐ€ RB0์„ ๊ณต๊ธ‰ํ•œ ์‹คํ—˜๊ตฌ๋ฅผ ์ œ์™ธํ•œ ๋ชจ๋“  ์‹คํ—˜๊ตฌ๋ณด๋‹ค ์œ ์˜์ ์œผ๋กœ ๋†’๊ฒŒ ๋‚˜ํƒ€๋‚ฌ๋‹ค. ๋˜ํ•œ RB0๊ณผ RB100์„ ๊ณต๊ธ‰ํ•œ ์‹คํ—˜๊ตฌ ์‚ฌ์ด์—๋Š” ์œ ์˜์ ์ธ ์ฐจ์ด๊ฐ€ ๋‚˜ํƒ€๋‚˜์ง€ ์•Š์•˜๋‹ค. ์ผ์ผ์„ฑ์žฅ๋ฅ (Specific growth rate, SGR)์€ RB40์„ ๊ณต๊ธ‰ํ•œ ์‹คํ—˜๊ตฌ๊ฐ€ ๋‹ค๋ฅธ ๋ชจ๋“  ์‹คํ—˜ ์‚ฌ๋ฃŒ๋ฅผ ๊ณต๊ธ‰ํ•œ ์‹คํ—˜๊ตฌ๋ณด๋‹ค ์œ ์˜์ ์œผ๋กœ ๋†’๊ฒŒ ๋‚˜ํƒ€๋‚ฌ๋‹ค. ๋˜ํ•œ RB0์„ ๊ณต๊ธ‰ํ•œ ์‹คํ—˜๊ตฌ์˜ ์ผ์ผ์„ฑ์žฅ๋ฅ ์€ RB100์„ ๊ณต๊ธ‰ํ•œ ์‹คํ—˜๊ตฌ๋ณด๋‹ค ์œ ์˜์ ์œผ๋กœ ๋†’๊ฒŒ ๋‚˜ํƒ€๋‚ฌ์œผ๋‚˜, RB20, RB60 ๋ฐ RB80์„ ๊ณต๊ธ‰ํ•œ ์‹คํ—˜๊ตฌ์™€๋Š” ์œ ์˜์ ์ธ ์ฐจ์ด๊ฐ€ ๋‚˜ํƒ€๋‚˜์ง€ ์•Š์•˜๋‹ค. ์ „๋ณต ๊ฐ€์‹๋ถ€์˜ ์ˆ˜๋ถ„, ์กฐ๋‹จ๋ฐฑ์งˆ, ํšŒ๋ถ„ ํ•จ๋Ÿ‰์€ ์‹คํ—˜์šฉ ๋ฐฐํ•ฉ์‚ฌ๋ฃŒ์— ์˜ํ•ด ์˜ํ–ฅ์„ ๋ฐ›์€ ๊ฒƒ์œผ๋กœ ๋‚˜ํƒ€๋‚ฌ๋‹ค. ๋ณธ ์—ฐ๊ตฌ ๊ฒฐ๊ณผ ์ „๋ณต์šฉ ๋ฐฐํ•ฉ์‚ฌ๋ฃŒ๋‚ด ๋‹ค์‹œ๋งˆ๋ฅผ 20% ์ฒจ๊ฐ€์‹œ ์ „๋ณต์˜ ์ฆ์ฒด๋Ÿ‰์— ๋ถ€์ •์ ์ธ ์˜ํ–ฅ ์—†์ด ๋‹ค์‹œ๋งˆ๋ฅผ ์ƒ๋ฏธ๊ฐ•์œผ๋กœ 100% ๊นŒ์ง€ ์™„์ „ํžˆ ๋Œ€์ฒด ๊ฐ€๋Šฅํ•˜๋ฉฐ, ๋˜ํ•œ ๋‹ค์‹œ๋งˆ๋ฅผ ์ƒ๋ฏธ๊ฐ•์œผ๋กœ 40% ๋Œ€์ฒด์‹œ ๊ฐ€์žฅ ์šฐ์ˆ˜ํ•œ ์ „๋ณต์˜ ์„ฑ์žฅ๋ฅ ์„ ๋ณด์˜€๋‹ค.Dietary substitution effect of sea tangle (ST) with rice bran (RB) on growth and carcass composition of juvenile abalone, Haliotis discus was determined. Juvenile abalone was acclimated to the experimental conditions for 4 weeks. Seventy Juvenile abalone averaging 0.43 g were randomly distributed into each of the 18, 70 L plastic rectangular containers. The experimental diets were fed to abalone once a day at a satiation level with a little leftover. The feeding trial lasted for 16 weeks. Survival of abalone was not affected by dietary substitution of ST with RB. However, weight gain of abalone fed the RB40 diet was higher than that of abalone fed the all other diets except for the RB0 diet. No significant difference in weight gain was found in abalone fed between the RB0 and RB100 diets. SGR of abalone fed the RB40 diet was higher than that of abalone fed all other diets. Also SGR of abalone fed the RB0 diet was higher than that of abalone fed the RB100 diet, but not different from that of abalone fed the RB20, RB60 and RB80 diets. Moisture, crude protein and ash content of the soft body of abalone were affected by dietary substitution of ST with RB. In conclusion, the 100% ST could be substituted with RB without a retardation of weight gain of abalone when the 20% ST was included into the experimental diet. The best growth performance was obtained in abalone fed the RB40 diet substituting 40% ST with RB.CONTENTS Contents i List of Tables ii Abstract (in korean) iii Abstract v I. Experiment 1 1. Introduction 1 2. Materials and Methods 3 2.1. Preparation of Abalone and Rearing Condition 3 2.2. Preparation of the Experimental Diets 3 2.3. Analytical Procedures of the Diets and Carcass 7 2.4. Statistical Analysis 7 3. Results 9 4. Discussion 13 II. Conclusion 17 III. Acknowledgements 18 IV. References 1

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    The effect of parents' Korean proficiency on their children's psychological health within multicultural families: Focusing on the mediated effect of the communication level between parents and their children

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    ๋ณธ ์—ฐ๊ตฌ๋Š” ๋‹ค๋ฌธํ™” ๊ฐ€์ •์ด ๊ฒช๋Š” ๊ณ ์œ ํ•œ ๋ฌธ์ œ ์ค‘ ํ•˜๋‚˜์ธ ๋ถ€๋ชจ์˜ ํ•œ๊ตญ์–ด ๋Šฅ๋ ฅ์œผ๋กœ ํŒŒ์ƒ๋˜๋Š” ํ˜„์ƒ์„ ๋ถ„์„ํ•˜๋Š” ์—ฐ๊ตฌ๋กœ์จ, ์–ด๋จธ๋‹ˆ์˜ ํ•œ๊ตญ์–ด ๋Šฅ๋ ฅ์ด ์ž๋…€์™€์˜ ์˜์‚ฌ์†Œํ†ต ๋ฐ ์ž๋…€์˜ ์‹ฌ๋ฆฌ์  ๊ฑด๊ฐ•์— ๋Œ€ํ•˜์—ฌ ์–ด๋– ํ•œ ์˜ํ–ฅ์„ ๋ฏธ์น˜๋Š”์ง€์— ๋Œ€ํ•˜์—ฌ ์ง‘์ค‘ํ•˜์—ฌ ๋ถ„์„ํ•œ๋‹ค. ํ•œ๊ตญ์ฒญ์†Œ๋…„์ •์ฑ…์—ฐ๊ตฌ์›์—์„œ ์ˆ˜ํ–‰ํ•œ ใ€Œ๋‹ค๋ฌธํ™”์ฒญ์†Œ๋…„ ํŒจ๋„์กฐ์‚ฌใ€์˜ 2์ฐจ(2012๋…„)~ 8์ฐจ(2018๋…„) ๋ฐ์ดํ„ฐ๋ฅผ ํ™œ์šฉํ•˜์—ฌ ๋ถ„์„์„ ์ง„ํ–‰ํ•˜์˜€๋‹ค. ๋จผ์ €, ๋Œ€์ƒ์ž๋กœ ์„ ์ •๋œ ์ „์ฒด ๋ฐ์ดํ„ฐ์˜ ํŠน์„ฑ์„ ํŒŒ์•…ํ•˜๊ธฐ ์œ„ํ•˜์—ฌ ์ง‘๋‹จ์ค‘์‹ฌ์ถ”์„ธ๋ชจํ˜•(Group Based Trajectory Modeling)์„ ๋ฐ”ํƒ•์œผ๋กœ ๊ธฐ์ˆ ํ†ต๊ณ„๋Ÿ‰์„ ๋ถ„์„ํ•˜์˜€๋‹ค. ๊ทธ๋ฆฌ๊ณ  ์–ด๋จธ๋‹ˆ์˜ ํ•œ๊ตญ์–ด ์ˆ˜์ค€๊ณผ ์ž๋…€์™€์˜ ์˜์‚ฌ์†Œํ†ต ์ˆ˜์ค€, ๊ทธ๋ฆฌ๊ณ  ์–ด๋จธ๋‹ˆ์˜ ํ•œ๊ตญ์–ด ์ˆ˜์ค€ ๋ฐ ์ž๋…€์™€์˜ ์˜์‚ฌ์†Œํ†ต ์ˆ˜์ค€๊ณผ ์ž๋…€์˜ ์šฐ์šธ ์ˆ˜์ค€์˜ ๊ด€๊ณ„์„ฑ์„ ๋ถ„์„ํ•˜๊ธฐ ์œ„ํ•˜์—ฌ ์„ ํ˜•ํ˜ผํ•ฉ๋ชจํ˜•(Linear Mixed Model)์„ ํ™œ์šฉํ•˜์—ฌ ๋ถ„์„ํ•˜์˜€์œผ๋ฉฐ ๋ถ€๋ชจ์™€ ์ž๋…€์˜ ์˜์‚ฌ์†Œํ†ต ์ˆ˜์ค€์˜ ๋งค๊ฐœ ์—ญํ• ์„ ๋ถ„์„ํ•˜๊ธฐ ์œ„ํ•ด Sobel test ๋ฐ Bootstrapping์„ ์ด์šฉํ•˜์—ฌ ๋งค๊ฐœ์š”์ธ ๋ถ„์„์„ ์‹œํ–‰ํ•˜์˜€๋‹ค. ๋ถ„์„์€ SAS version 9.4 ํ†ต๊ณ„ ํ”„๋กœ๊ทธ๋žจ์„ ์ด์šฉํ•˜์˜€์œผ๋ฉฐ PROC TRAJ, PROC MIXED, PROC CAUSALMED ๋“ฑ์„ ์‚ฌ์šฉํ•˜์˜€๋‹ค. ์šฐ๋ฆฌ๋Š” ๋ณธ ๋ถ„์„์„ ํ†ตํ•˜์—ฌ ๋‹ค๋ฌธํ™” ๊ฐ€์ • ์–ด๋จธ๋‹ˆ์˜ ํ•œ๊ตญ์–ด ๋Šฅ๋ ฅ์˜ ๋ถ€์กฑ์ด ๋ถ€๋ชจ์™€ ์ž๋…€ ๊ฐ„์˜ ์˜์‚ฌ์†Œํ†ต ์ˆ˜์ค€์„ ์ €ํ•ดํ•˜๋Š” ์š”์†Œ๋กœ ์ž‘์šฉ์„ ํ•˜๊ฒŒ ๋จ์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ์—ˆ๋‹ค. ๋˜ํ•œ, ๋‹ค๋ฌธํ™” ๊ฐ€์ • ๋‚ด์—์„œ๋„ ๋ถ€๋ชจ์™€ ์ž๋…€ ๊ฐ„์˜ ์˜์‚ฌ์†Œํ†ต์˜ ์›ํ™œํ•œ ์˜์‚ฌ์†Œํ†ต์€ ์ž๋…€์˜ ์šฐ์šธ ์ˆ˜์ค€์„ ๋‚ฎ์ถ”๋Š” ๋ฐ ์˜ํ–ฅ์„ ์ค„ ์ˆ˜ ์žˆ์Œ์„ ๋ณด์˜€๋‹ค. ๊ทธ๋ฆฌ๊ณ  ์ด ๊ฒฐ๊ณผ๋ฅผ ๋ฐ”ํƒ•์œผ๋กœ ๋‹ค๋ฌธํ™” ๊ฐ€์ • ์–ด๋จธ๋‹ˆ์˜ ํ•œ๊ตญ์–ด ๋Šฅ๋ ฅ์ด ๋ถ€๋ชจ์™€ ์ž๋…€ ๊ฐ„์˜ ์˜์‚ฌ์†Œํ†ต ์ˆ˜์ค€์„ ๋งค๊ฐœ๋กœ ์ž๋…€์˜ ์šฐ์šธ ์ˆ˜์ค€์— ์˜ํ–ฅ์„ ๋ฏธ์นจ์„ ๋ณด์˜€๋‹ค. ๋ณธ ์—ฐ๊ตฌ์—์„œ ๋ณด์ธ ๊ฒฐ๊ณผ๋“ค์€ ๋งŽ์€ ๋‹ค๋ฌธํ™” ๊ฐ€์ •์ด ํ˜„์‹ค์—์„œ ๋ถ€๋”ชํžˆ๊ฒŒ ๋˜๋Š” ๊ฐ€์žฅ ์ค‘์š”ํ•œ ๋ฌธ์ œ์ธ ์–ธ์–ด๋ฌธ์ œ๋ฅผ ๊ณ ๋ คํ•˜์—ฌ ์ด๋กœ๋ถ€ํ„ฐ ํŒŒ์ƒ๋  ์ˆ˜ ์žˆ๋Š” ๋‹ค์–‘ํ•œ ํ˜„์ƒ ์ค‘, ์ฒญ์†Œ๋…„๊ธฐ์˜ ์ž๋…€์—๊ฒŒ ๋ฏธ์น˜๋Š” ์˜ํ–ฅ์„ ๋ถ„์„ํ–ˆ๋‹ค๋Š” ์ ์—์„œ ์˜์˜๊ฐ€ ์žˆ๋‹ค. ๋ณธ ์—ฐ๊ตฌ์˜ ๊ฒฐ๊ณผ๋Š” ๋‹ค๋ฌธํ™” ๊ฐ€์ • ์ง€์› ํ”„๋กœ๊ทธ๋žจ ๋“ฑ์„ ๊ตฌ์„ฑํ•˜๋Š” ์ฐจ์›์—์„œ ์—ฌ๋Ÿฌ ๊ฐ€์ง€ ํ˜„์‹ค์ ์ธ ์ œ์–ธ์œผ๋กœ ์ด์–ด์งˆ ์ˆ˜ ์žˆ๋‹ค๋Š” ์˜์˜๊ฐ€ ์žˆ๋‹ค. ํŠนํžˆ, ๋‹ค๋ฌธํ™” ๊ฐ€์ • ๋‚ด์—์„œ์˜ ๋ถ€๋ชจ์˜ ํ•œ๊ตญ์–ด ๋Šฅ๋ ฅ์ด ์ž๋…€๊ฐ€ ์ฒญ์†Œ๋…„๊ธฐ์— ์ ‘์–ด๋“  ์ดํ›„์—๋„ ์ž๋…€์—๊ฒŒ ์˜ํ–ฅ์„ ๋ฏธ์น˜๋Š” ๊ฒƒ์„ ๋ณด์ž„์œผ๋กœ์จ ๋‹ค๋ฌธํ™” ๊ฐ€์ • ๋ถ€๋ชจ๋“ค์—๊ฒŒ ์ง€์†์ ์ธ ์–ธ์–ด ๊ต์œก์„ ์ œ๊ณตํ•˜๋Š” ๊ฒƒ์— ๋Œ€ํ•œ ํ•„์š”์„ฑ์„ ์ œ์‹œํ•˜์˜€๋‹ค. ๋˜ํ•œ, ๋‹ค๋ฌธํ™” ๊ฐ€์ • ๋‚ด์—์„œ๋„ ๋ถ€๋ชจ์™€ ์ž๋…€ ๊ฐ„์˜ ์›ํ™œํ•œ ์˜์‚ฌ์†Œํ†ต์ด ๊ฐ€์ง€๋Š” ์ค‘์š”๋„๋ฅผ ๋ณด์ž„์œผ๋กœ์จ ๋ถ€๋ชจ์™€ ์ž๋…€ ๊ฐ„์˜ ์„œ๋กœ์˜ ์ฐจ์ด๋ฅผ ์ดํ•ดํ•  ์ˆ˜ ์žˆ๋Š” ๋ฌธํ™”๊ต์œก ๋“ฑ์„ ์ง€์›ํ•˜๋Š” ๊ฒƒ๊ณผ ๊ฐ™์€ ํ˜•ํƒœ๋กœ์จ ํ˜„์‹ค์ ์ธ ์ œ์–ธ์œผ๋กœ๋„ ์ด์–ด์งˆ ์ˆ˜ ์žˆ์„ ๊ฒƒ์œผ๋กœ ๊ธฐ๋Œ€๋œ๋‹ค. ์ด๋ฒˆ ๋ถ„์„์„ ๊ธฐ์ดˆ๋กœ ํ•˜์—ฌ ์ง€๊ธˆ๊นŒ์ง€ ์ˆ˜์ง‘๋œ ๋ฐ์ดํ„ฐ์˜ ๋‹ค์–‘ํ•œ ํ•ญ๋ชฉ์„ ํ™œ์šฉํ•œ ์—ฐ๊ตฌ๋กœ ํ™•์žฅํ•œ๋‹ค๋ฉด ๋‹ค๋ฌธํ™” ๊ฐ€์ •์ด ํ•œ๊ตญ ์‚ฌํšŒ์— ์ ์‘ํ•˜๋ฉฐ ์ƒ๊ธธ ์ˆ˜ ์žˆ๋Š” ์—ฌ๋Ÿฌ ๋ฌธ์ œ์— ๋Œ€ํ•œ ์‹œ์‚ฌ์ ์„ ์ œ๊ณตํ•  ์ˆ˜ ์žˆ๋Š” ์—ฐ๊ตฌ๋กœ ๋ฐœ์ „๋  ์ˆ˜ ์žˆ์„ ๊ฒƒ์œผ๋กœ ์ƒ๊ฐ๋œ๋‹ค. This study identifies phenomena derived from the lack of Korean proficiency of the parent who belongs to multicultural families, which is one of the unique problems experienced by them. In addition, we focus on how their Korean proficiency affects communication with their children and their psychological health. This study used Multicultural Adolescents Panel Study data of seven consecutive years from 2011(second survey data) to 2018(eighth survey data), which the National Youth Policy Institute conducts. We utilize SAS 9.4 for data analysis, and it used PROC TRAJ, PROC MIXED, PROC CAUSALMED, etc, for the SAS function. First, to organize descriptive statistics of the longitudinal data, we use the Group-Based Trajectory Modeling(GBTM). Then, we decompose three groups based on children's depression levels, and we find a tendency in mothers' Korean proficiency and communication level in all groups. Then, we use the Linear Mixed Model(LMM) to figure out the relationship between the mothers' Korean proficiency and communication level as well as the communication level and children's depression levels. Through this analysis, we find that parents' lack of Korean proficiency acted as a factor hindering the communication level between parents and children. Also, it shows that the smooth communication between parents and children affects lowering the children's depression level even within multicultural families. Lastly, to analyze the meditated effect of communication level between the parents' Korean proficiency and the children's depression level using the Sobel test and Bootstrapping. Through this, we reveal that communication level acted as a role of mediation between two aforementioned factors. Our results are significant in that we analyze the effects of language problems experienced by most multicultural families on their adolescent children. Our results can provide various practical suggestions for constructing a support program for multicultural families. In particular, we suggest the need to provide continuous language education to parents of multicultural families by showing that the Koren proficiency of parents affects their children even after they enter adolescence. In addition, our results also suggest the importance of cultural education programs to understand cultural differences between parents and children by showing the importance of communication between them. We expect that, based on our study, the further studies can provide various implications by considering other than depression level among psychological health. We believe that this study stream would help identify the several problems that multicultural families may face and help establish various practical suggestions that can adapt to Korean society.open์„

    ์ƒํ•˜์•… ์ œ1๋Œ€๊ตฌ์น˜ ๊ทผ์‹ฌํ˜‘์ธก์น˜๊ทผ๊ณผ ๊ทผ์‹ฌ์น˜๊ทผ์—์„œ์˜ ๋ฏธ์„ธ์ „์‚ฐํ™” ๋‹จ์ธต์ดฌ์˜๊ณผ ์น˜์•„ํˆฌ๋ช…ํ™” ๋ฐฉ๋ฒ•์„ ์ด์šฉํ•œ ๋ฏธ์„ธ ๊ทผ๊ด€ ํ˜•ํƒœ ๋น„๊ต

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    ํ•™์œ„๋…ผ๋ฌธ (๋ฐ•์‚ฌ)-- ์„œ์šธ๋Œ€ํ•™๊ต ๋Œ€ํ•™์› : ์น˜์˜๊ณผํ•™๊ณผ, 2015. 2. ๊ธˆ๊ธฐ์—ฐ.Objectives. Micro-computed tomography (MCT) with alternative image reformatting techniques shows complex and detailed root canal anatomy. This study compared the MCT images reconstructed by two-dimensional (2D) thin-slab minimum intensity projection (TS-MinIP) and three-dimensional (3D) volume-rendering technique with the images obtained from the clearing technique, to study detailed root canal morphology in maxillary first molar mesiobuccal (MB) roots and mandibular first molar mesial (MS) roots. Methods. Extracted human maxillary first molar MB roots (n = 18) and mandibular first molar MS roots (n = 31) were scanned by MCT (Skyscan 1172). The MCT images were constructed using 2D TS-MinIP and 3D volume-rendering technique. The same teeth were then processed by the clearing technique and the clearing images were obtained. For each root, the clearing, the 2D TS-MinIP, the 3D volume-rendered, and the combined 2D TS-MinIP and 3D volume-rendered images were examined independently by 4 dentists and categorized according to Vertuccis classification. Fine anatomical structures such as accessory canals, intercanal communications, and loops were also compared. Results. Complex canal systems were more clearly visible in the MCT images than the clearing images. On each root, the 3 MCT images showed the same canal configuration, whereas the clearing images showed less complicated configuration than the MCT ones. The frequency of non-classifiable configurations by Vertuccis classification was shown as 38.9% in maxillary first molar MB roots and 51.6% in mandibular first molar MS roots. Fine anatomical structures such as intercanal communications, accessory canals, and loops were mostly found with the combined 2D TS-MinIP and 3D volume-rendered images. Conclusions. The combined 2D TS-MinIP and 3D volume-rendered images of MCT could be useful for morphological study of complex root canal systems. In the future, the establishment of a systematic classification that embraces non-classifiable canal configurations found in this study may help the root canal morphological study.Abstract I. Introduction II. Materials and Methods III. Results IV. Discussion V. References Tables and Figures Abstract in KoreanDocto

    ์ž์—ฐ๊ฒฝ๊ด€ ๊ฐ์ƒ์ด ์ž์—ฐ ์œ ๋Œ€๊ฐ๊ณผ ์‚ฌํšŒ์  ๊ฐ€์น˜์ง€ํ–ฅ์„ฑ์— ๋ฏธ์น˜๋Š” ์˜ํ–ฅ

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    ํ•™์œ„๋…ผ๋ฌธ (์„์‚ฌ)-- ์„œ์šธ๋Œ€ํ•™๊ต ๋Œ€ํ•™์› : ํ™˜๊ฒฝ๊ต์œก์ „๊ณต, 2016. 8. ๋ฅ˜์žฌ๋ช….๋ฐ”์œ ์ƒํ™œ ์†์—์„œ ์ž์—ฐ๊ณผ ๋‹จ์ ˆ๋˜์–ด ์‚ด์•„๊ฐ€๋Š” ํ˜„๋Œ€์ธ๋“ค์˜ ์ƒํ™œ์Šต๊ด€์€ ์‚ฌ๋žŒ๋“ค์ด ์ž์—ฐํ™˜๊ฒฝ์„ ์ธ์‹ํ•˜๋Š” ๋ฐฉ์‹์—๋„ ์˜ํ–ฅ์„ ๋ฏธ์ณ ์ •์„œ์ ์œผ๋กœ ์ž์‹ ์ด ์ƒํƒœ๊ณ„์˜ ์ผ์›์ด๋ผ๋Š” ๋Š๋‚Œ์„ ๊ฐ–๊ธฐ ์–ด๋ ต๊ฒŒ ๋˜์—ˆ๋‹ค. ๋‚˜์™€ ์ž์—ฐ ์‚ฌ์ด์˜ ์—ฐ๊ฒฐ๊ณ ๋ฆฌ๋ฅผ ์žƒ์–ด๋ฒ„๋ฆฐ ์‚ฌ๋žŒ๋“ค์€ ๋” ์ด์ƒ ๋ถˆํŽธํ•จ์„ ๊ฐ์ˆ˜ํ•˜๊ณ  ์นœํ™˜๊ฒฝ์ ์ธ ํ–‰๋™์„ ์‹ค์ฒœํ•˜๊ณ ์ž ํ•˜๋Š” ์˜์ง€๋ฅผ ๋ณด์ด์ง€ ์•Š๊ฒŒ ๋˜์—ˆ๋‹ค. ๋˜ํ•œ ๋‹จ์‹œ๊ฐ„์— ์–ป์„ ์ˆ˜ ์žˆ๋Š” ๊ฐœ์ธ์˜ ์ด์ต๊ณผ ๋‹น์žฅ์˜ ๊ฐœ์ธ์  ์ด์ต์„ ๋‹ค์†Œ ํฌ์ƒํ•˜๊ณ  ์ง‘๋‹จ ๊ตฌ์„ฑ์›์˜ ํ˜‘๋™์œผ๋กœ ์–ป์„ ์ˆ˜ ์žˆ๋Š” ์ง‘๋‹จ์˜ ์ด์ต์ด ์ƒ์ถฉํ•˜๋Š” ํ™˜๊ฒฝ๋ฌธ์ œ์— ์žˆ์–ด ๊ฐœ์ธ์˜ ์ด์ต์„ ์ถ”๊ตฌํ•˜๋Š” ์‚ฌ๋žŒ๋“ค์ด ๋Š˜์–ด๋‚˜๊ฒŒ ๋˜์—ˆ๋‹ค. ๊ทธ ๊ฒฐ๊ณผ ๊ธฐํ›„๋ณ€ํ™”, ๊ฐ์ข… ํ™˜๊ฒฝ์˜ค์—ผ, ๋ฉธ์ข… ์œ„๊ธฐ์ข… ์ฆ๊ฐ€์™€ ๊ฐ™์€ ์ƒํƒœ์  ์œ„๊ธฐ๊ฐ€ ์ดˆ๋ž˜ํ•˜์˜€๋‹ค. ์ผ์ƒ์˜ ๋Œ€๋ถ€๋ถ„์„ ์ž์—ฐ๊ณผ๋Š” ๋–จ์–ด์ง„ ์‹ค๋‚ด์—์„œ ๋ณด๋‚ด๋Š” ํ˜„๋Œ€์ธ๋“ค์˜ ์ƒํ™œ์Šต๊ด€์—์„œ ์šฐ๋ฆฌ๋‚˜๋ผ์˜ ์ฒญ์†Œ๋…„๋“ค๋„ ์ž์œ ๋กœ์šธ ์ˆ˜๋Š” ์—†๋‹ค. ์ด์— ๋ณธ ์—ฐ๊ตฌ์—์„œ๋Š” ํ™˜๊ฒฝ์ ์œผ๋กœ ์˜๋ฏธ ์žˆ๋Š” ์ง์ ‘์ฒดํ—˜์˜ ๊ธฐํšŒ๋ฅผ ๊ฐ–๊ธฐ ์–ด๋ ค์šด ๊ต์œกํ˜„์žฅ์˜ ํ•œ๊ณ„๋ฅผ ๊ทน๋ณตํ•˜๊ณ  ํ•™์ƒ๋“ค์—๊ฒŒ ์ƒ๋™๊ฐ ์žˆ๊ณ  ์ง€์†์ ์ธ ์ž์—ฐ์ฒดํ—˜์˜ ๊ธฐํšŒ๋ฅผ ์ œ๊ณตํ•˜๋Š” ๋ฐฉ์•ˆ์œผ๋กœ ์˜์ƒ์„ ํ™œ์šฉํ•œ ์ž์—ฐ ํ™˜๊ฒฝ์˜ ๊ฐ„์ ‘์ฒดํ—˜์„ ์ œ์‹œํ•˜์˜€๋‹ค. ์˜์ƒ์„ ํ†ตํ•œ ์ž์—ฐ๊ฒฝ๊ด€์˜ ๊ฐ์ƒ ๊ฒฝํ—˜์ด ํ•™์ƒ๋“ค์˜ ์นœํ™˜๊ฒฝ์  ์ •์„œ ํ˜•์„ฑ์— ๊ธ์ •์ ์ธ ์˜ํ–ฅ์„ ๋ฏธ์น˜๊ณ  ์ด๊ฒƒ์ด ๊ฐœ์ธ์˜ ์ด์ต์ถ”๊ตฌ์™€ ํƒ€์ธ๊ณผ์˜ ํ˜‘๋ ฅ์ด ์š”๊ตฌ๋˜๋Š” ๋”œ๋ ˆ๋งˆ ์ƒํ™ฉ์—์„œ ํ˜‘๋ ฅ์  ํƒœ๋„๋ฅผ ๋ณด์ด๋Š”๋ฐ ๋„์›€์„ ์ค„ ๊ฒƒ์ด๋ผ๋Š” ๊ฐ€์„ค์„ ์„ธ์šฐ๊ณ  ์ด๋ฅผ ์‹คํ—˜์—ฐ๊ตฌ๋ฅผ ํ†ตํ•ด ์ฆ๋ช…ํ•˜๊ณ ์ž ํ•˜์˜€๋‹ค. ์„œ์šธ์— ๊ฑฐ์ฃผํ•˜๊ณ  ์žˆ๋Š” ์ดˆ๋“ฑํ•™๊ต 5,6ํ•™๋…„ ํ•™์ƒ 303๋ช…(๋‚จ: 160๋ช…, ์—ฌ: 143๋ช…)์„ ์‹คํ—˜ ๋Œ€์ƒ์œผ๋กœ ์„ ์ •ํ•˜์˜€๋‹ค. ํ•ด๋‹น ํ•™๋ น์˜ ํ•™์ƒ๋“ค์ด ์ž์—ฐ ๋ฐ ์ธ๊ณตํ™˜๊ฒฝ์„ ๊ตฌ์„ฑํ•˜๋Š” ์š”์†Œ์— ๋Œ€ํ•ด ์–ด๋– ํ•œ ์ธ์‹์„ ๊ฐ–๊ณ  ์žˆ๋Š”์ง€ ์„ ํ–‰์—ฐ๊ตฌ ๊ฒฐ๊ณผ์— ๊ทผ๊ฑฐํ•˜์—ฌ ์ž์—ฐ๊ฒฝ๊ด€ ์˜์ƒ๊ณผ ์ธ๊ณต๊ฒฝ๊ด€ ์˜์ƒ์˜ ๋‚ด์šฉ์„ ๊ตฌ์„ฑํ•˜์˜€๋‹ค. ์„ ํ–‰์—ฐ๊ตฌ์˜ ๊ณ ์ฐฐ์„ ํ†ตํ•ด ์ •์„œ์  ์ธก๋ฉด์˜ ๋ณ€ํ™”๋ฅผ ์‚ดํŽด๋ณผ ๋ณ€์ˆ˜๋กœ ์ž์—ฐ์œ ๋Œ€๊ฐ์„, ํ˜‘๋ ฅ์  ํƒœ๋„ ์ธก๋ฉด์˜ ๋ณ€์ˆ˜๋Š” ์‚ฌํšŒ์  ๊ฐ€์น˜์ง€ํ–ฅ์„ฑ์„ ์„ค์ •ํ•˜๊ณ , ๊ฐ ๋ณ€์ˆ˜๋ฅผ ์ธก์ •ํ•˜๊ธฐ ์œ„ํ•œ ์„ค๋ฌธ์ง€๋ฅผ ๊ฐœ๋ฐœํ•˜์˜€๋‹ค. ์ž์—ฐํ™˜๊ฒฝ ์š”์†Œ๋กœ๋งŒ ์ด๋ฃจ์–ด์ง„ ์˜์ƒ์„ ๊ฐ์ƒํ•œ ์‹คํ—˜์ง‘๋‹จ๊ณผ ์ž์—ฐ๊ณผ ๋Œ€์กฐ๋˜๋Š” ์ธ๊ณตํ™˜๊ฒฝ ์š”์†Œ๋กœ๋งŒ ์ด๋ฃจ์–ด์ง„ ์˜์ƒ์„ ๋ณธ ๋น„๊ต์ง‘๋‹จ์˜ ์ž์—ฐ ์œ ๋Œ€๊ฐ ๋ฐ ์‚ฌํšŒ์  ๊ฐ€์น˜์ง€ํ–ฅ์„ฑ์„ ๊ฐ๊ฐ ์ธก์ •ํ•˜์—ฌ ๊ทธ ๊ฒฐ๊ณผ๋ฅผ ๋น„๊ตํ•˜์˜€๋‹ค. ์ตœ๋Œ€ํ•œ ๋ฌด์„ ํ‘œ์ง‘ ์ƒํ™ฉ๊ณผ ๊ทผ์ ‘ํ•œ ์กฐ๊ฑด ํ˜•์„ฑ์„ ์œ„ํ•ด ์‹คํ—˜์— ์ฐธ์—ฌํ•œ ํ•™์ƒ๋“ค์—๊ฒŒ ๊ฐ๊ฐ ๋‹ค๋ฅธ ์ข…๋ฅ˜์˜ ์˜์ƒ์ด ๋‹ด๊ฒจ์žˆ๋Š” ํƒœ๋ธ”๋ฆฟ PC๋ฅผ ๋ฌด์ž‘์œ„๋กœ ๋ฐฐํฌํ•˜์—ฌ ์ž์—ฐ ๋ฐ ์ธ๊ณต๊ฒฝ๊ด€์„ ๊ฐ์ƒํ•  ์ˆ˜ ์žˆ๋„๋ก ํ•œ ํ›„ ์ž์—ฐ์œ ๋Œ€๊ฐ๊ณผ ์‚ฌํšŒ์  ๊ฐ€์น˜์ง€ํ–ฅ์„ฑ์„ ์ธก์ •ํ•˜๋Š” ์„ค๋ฌธ์ง€๋ฅผ ๋ฐฐ๋ถ€ํ•˜์˜€๋‹ค. ๊ทธ ๊ฒฐ๊ณผ ์‹คํ—˜์ง‘๋‹จ์ด์—ˆ๋˜ ์ž์—ฐ๊ฒฝ๊ด€์„ ๊ฐ์ƒํ•œ ์ดˆ๋“ฑํ•™์ƒ๋“ค์˜ ์ž์—ฐ์œ ๋Œ€๊ฐ๊ณผ ์‚ฌํšŒ์  ๊ฐ€์น˜์ง€ํ–ฅ์„ฑ์ด ๋น„๊ต์ง‘๋‹จ์˜ ๊ทธ๊ฒƒ์— ๋น„ํ•ด ๋ชจ๋‘ ์œ ์˜์ˆ˜์ค€ .001์—์„œ ์œ ์˜๋ฏธํ•˜๊ฒŒ ๋†’์Œ์„ ์•Œ ์ˆ˜ ์žˆ์—ˆ๋‹ค. ๋˜ํ•œ ์ž์—ฐ์œ ๋Œ€๊ฐ๊ณผ ์‚ฌํšŒ์  ๊ฐ€์น˜์ง€ํ–ฅ์„ฑ ์‚ฌ์ด์˜ ์ƒ๊ด€๊ด€๊ณ„๋ฅผ ์‚ฐ์ถœํ•œ ๊ฒฐ๊ณผ .000์˜ ์œ ์˜์ˆ˜์ค€์—์„œ ๋‹ค์†Œ ๋†’์€ ์–‘์˜ ์ƒ๊ด€๊ด€๊ณ„๊ฐ€ ์žˆ์Œ์ด ํ™•์ธ๋˜์—ˆ๋‹ค. ์ด๋Š” ์ž์—ฐ๊ฒฝ๊ด€์˜ ๊ฐ์ƒ์ด ํ•™์ƒ๋“ค์˜ ์นœํ™˜๊ฒฝ์  ์ •์„œ์™€ ํ˜‘๋ ฅ์  ํƒœ๋„์— ๊ธ์ •์ ์ธ ์˜ํ–ฅ์„ ๋ฏธ์นœ๋‹ค๋Š” ์‚ฌ์‹ค์„ ๋ณด์—ฌ์ค€๋‹ค. ๋‚˜์•„๊ฐ€ ์ž์—ฐ์œ ๋Œ€๊ฐ์ด ๋†’์€ ์‚ฌ๋žŒ์€ ํ˜‘๋ ฅ์  ์„ฑํ–ฅ์ด ๊ฐ•ํ•œ ์‚ฌ๋žŒ์œผ๋กœ์„œ ํ™˜๊ฒฝ๋ฌธ์ œ์— ์žˆ์–ด ํ™˜๊ฒฝ์„ ๋ณดํ˜ธํ•˜๋Š” ์„ ํƒ์„ ํ•  ๊ฒƒ์œผ๋กœ, ํ˜‘๋ ฅ ์„ฑํ–ฅ์ด ๊ฐ•ํ•œ ์‚ฌ๋žŒ์€ ์ž์—ฐ ์œ ๋Œ€๊ฐ ๋˜ํ•œ ๊ฐ•ํ•œ ์‚ฌ๋žŒ์ผ ํ™•๋ฅ ์ด ๋†’์„ ๊ฒƒ์œผ๋กœ ์˜ˆ์ธกํ•ด๋ณผ ์ˆ˜ ์žˆ๋‹ค. ์ด ์—ฐ๊ตฌ๋Š” ์ž์—ฐ์— ๋Œ€ํ•œ ๋‹จ์ˆœํ•œ ๊ฐ„์ ‘์ฒดํ—˜๋„ ํ•™์ƒ๋“ค์ด ์ •์„œ์ ์œผ๋กœ ์ž์—ฐ์„ ๊ฐ€๊น๊ฒŒ ๋Š๋ผ๋Š”๋ฐ ๋„์›€์„ ์ฃผ๊ณ , ํ˜‘๋ ฅ์  ํ–‰๋™์„ฑํ–ฅ์—๋„ ๊ธ์ •์ ์ธ ์˜ํ–ฅ์„ ๋ฏธ์ณ ํ™˜๊ฒฝ์นœํ™”์  ์„ ํƒ์„ ํ•˜๋Š”๋ฐ ๋„์›€์„ ์ค„ ์ˆ˜ ์žˆ๋‹ค๋Š” ์‚ฌ์‹ค์„ ๋ฐํ˜”๋‹ค๋Š” ์ ์—์„œ ์˜๋ฏธ๊ฐ€ ์žˆ๋‹ค. ๋‚˜์•„๊ฐ€ ํ˜‘๋ ฅ์  ํ–‰๋™ ์„ฑํ–ฅ์€ ํ˜„ ๊ต์œกํ˜„์žฅ์—์„œ ์ค‘์ ์ ์œผ๋กœ ์ง€๋„ํ•˜๋Š” ์ธ์„ฑ๊ต์œก๊ณผ๋„ ์—ฐ๊ด€๋˜๋Š” ๋ถ€๋ถ„์œผ๋กœ ์ž์—ฐ ๊ฐ์ƒ์˜ ์ธ์„ฑ ๊ต์œก์  ํšจ๊ณผ๊นŒ์ง€๋„ ํ™•์ธํ•  ์ˆ˜ ์žˆ๋‹ค๋Š” ์ ์—์„œ ๋”์šฑ ์˜๋ฏธ๊ฐ€ ๊นŠ๋‹ค. ๋ณธ ์—ฐ๊ตฌ์˜ ๊ฒฐ๊ณผ๋ฅผ ๋ฐ”ํƒ•์œผ๋กœ ๋‹ค์–‘ํ•œ ์ˆ˜์—… ํ™œ๋™ ์ƒํ™ฉ์—์„œ ํ•™์ƒ๋“ค์ด ์ž์—ฐ ๊ฒฝ๊ด€์„ ๊ฐ์ƒํ•  ์ˆ˜ ์žˆ๋Š” ๊ธฐํšŒ๊ฐ€ ๋Š˜์–ด๋‚˜๊ณ  ํ™˜๊ฒฝ๊ต์œก์˜ ์ธ์„ฑ ๊ต์œก์  ํšจ๊ณผ์— ๋Œ€ํ•œ ๋…ผ์˜๊ฐ€ ์ด์–ด์งˆ ์ˆ˜ ์žˆ๊ธฐ๋ฅผ ๊ธฐ๋Œ€ํ•œ๋‹ค.์ œ 1 ์žฅ. ์„œ๋ก  1 ์ œ 1 ์ ˆ. ์—ฐ๊ตฌ์˜ ๋ฐฐ๊ฒฝ ๋ฐ ํ•„์š”์„ฑ 1 ์ œ 2 ์ ˆ. ์—ฐ๊ตฌ๋ชฉ์  ๋ฐ ์—ฐ๊ตฌ๋ฌธ์ œ 4 ์ œ 2 ์žฅ. ์ด๋ก ์  ๋ฐฐ๊ฒฝ 5 ์ œ 1 ์ ˆ. ์ž์—ฐ๊ฒฝ๊ด€ 5 1. ์ž์—ฐ๊ฒฝ๊ด€์˜ ๊ฐœ๋… 5 2. ์ž์—ฐ๊ฒฝ๊ด€ ๊ฐ์ƒ์˜ ์‹ฌ๋ฆฌ์  ํšจ๊ณผ 6 ์ œ 2 ์ ˆ. ์ž์—ฐ์œ ๋Œ€๊ฐ 8 1. ์ž์—ฐ์œ ๋Œ€๊ฐ์˜ ๊ฐœ๋… 8 2. ์ž์—ฐ์œ ๋Œ€๊ฐ์˜ ์ธก์ • 9 ์ œ 3 ์ ˆ. ์‚ฌํšŒ์  ๊ฐ€์น˜์ง€ํ–ฅ์„ฑ 11 1. ์‚ฌํšŒ์  ๊ฐ€์น˜์ง€ํ–ฅ์„ฑ์˜ ๊ฐœ๋… 11 2. ์‚ฌํšŒ์  ๊ฐ€์น˜์ง€ํ–ฅ์„ฑ์˜ ํ™˜๊ฒฝ๊ต์œก์  ์˜๋ฏธ 13 3. ์‚ฌํšŒ์  ๊ฐ€์น˜์ง€ํ–ฅ์„ฑ์˜ ์ธก์ • 15 ์ œ 3 ์žฅ. ์—ฐ๊ตฌ๋ฐฉ๋ฒ• 17 ์ œ 1 ์ ˆ. ์‹คํ—˜์„ค๊ณ„ 18 1. ์—ฐ๊ตฌ๊ฐ€์„ค 18 2. ์—ฐ๊ตฌ๋Œ€์ƒ 19 3. ์‹คํ—˜์ฒ˜์น˜ ์ž๋ฃŒ ๋ฐ ๋ฐฉ๋ฒ• 19 4. ์ธก์ •๋„๊ตฌ 21 ์ œ 2 ์ ˆ. ์‹คํ—˜์ ˆ์ฐจ 27 ์ œ 3 ์ ˆ. ์ž๋ฃŒ ๋ฐ ๊ฒฐ๊ณผ์ฒ˜๋ฆฌ 27 ์ œ 4 ์žฅ. ์—ฐ๊ตฌ ๊ฒฐ๊ณผ 28 ์ œ 1 ์ ˆ. ๊ฒฝ๊ด€์˜ ์ข…๋ฅ˜์™€ ์ž์—ฐ์œ ๋Œ€๊ฐ 28 ์ œ 2 ์ ˆ. ๊ฒฝ๊ด€์˜ ์ข…๋ฅ˜์™€ ์‚ฌํšŒ์  ๊ฐ€์น˜์ง€ํ–ฅ์„ฑ 31 ์ œ 3 ์ ˆ. ์ž์—ฐ์œ ๋Œ€๊ฐ๊ณผ ์‚ฌํšŒ์  ๊ฐ€์น˜์ง€ํ–ฅ์„ฑ 34 ์ œ 5 ์žฅ. ๋…ผ์˜ ๋ฐ ๊ฒฐ๋ก  36 ์ œ 1 ์ ˆ. ์ข…ํ•ฉ ๋…ผ์˜ ๋ฐ ๊ฒฐ๋ก  36 ์ œ 2 ์ ˆ. ์—ฐ๊ตฌ์˜ ์˜์˜ ๋ฐ ์ œ์–ธ 38 ์ฐธ๊ณ ๋ฌธํ—Œ 41 Abstract 53Maste

    In vitro and in vivo optimization of Destabilizing Domain in neuron for modeling brain disease

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    The destabilizing domain (DD) from Escherichia coli dihydrofolate reductase (ecDHFR) is a relatively small protein (158 a.a), which can conditionally control the abundance of a protein of interests (POIs) by a molecular chaperone, trimethoprim (TMP). The DD, allowed an ideal approach for reversibly and repeatedly regulating protein stability, has been used successfully to control the protein level and potentially activity. Therefore, we aim to test the usability of the DD for selectively regulating protein stability in neurons in vivo and in vitro. First, we utilized the PiggyBac (PB) transposon system for moderate and permanent expressions of candidate genes by genome integration. Using in utero electroporation (IUE), we introduced DD-YFP to developing ventricle at embryonic day 15.5 followed by TMP administration at postnatal day 14. Strong YFP fluorescence was detected 6hr after the administration of TMP whereas minimal or no YFP fluorescence was detected in untreated group suggesting effective stabilization of DD-YFP by TMP and minimal leakage in the absence of TMP. Currently, we are generating stable human embryonic stem cell (hESC) lines expressing DD-tagged GFP followed by neuronal differentiation to validate the TMP-induced stabilization of GFP in vitro. Altogether, our study will reveal the usability of the DD for selectively regulating protein stability in neurons in vivo and in vitro.1

    Pathogenic Mechanism of Epilepsy in Sebaceous Nevus Syndrome by Dysregulation of RAS/MAPK Pathway

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    RAS/MAPK pathway involves not only biological processes such as differentiation, proliferation, and migration but also many diseases such as cancer. RASopathies, which are a group of developmental disorders caused by mutation of the RAS/MAPK pathway component, suggest the crucial role of this pathway in normal development. Sebaceous nevus syndrome (SNS), one of the RASopathies, is caused by somatic gain-of-function mutation of HRAS or KRAS. The symptoms of SNS include central-nervous system-related defects such as cerebral defects, intellectual disability, and epilepsy. To examine the pathogenesis of neurological symptoms in SNS, we generated the disease mouse model by overexpression of KRAS p.G12V in developing cortex using in utero electroporation. This model successfully recapitulated histological manifestations observed in the patient, such as heterotopic neurons, gliosis, and hypomyelination. It also showed abnormal neuronal differentiation and migration and neuronal dysmorphogenesis. To investigate neuronal excitability at a cellular level, we analyzed electrophysiological properties using patch-clamp. Both neurons with and without KRAS p.G12V overexpression showed hyperexcitability. Interneurons also showed abnormal action potential properties. To elucidate the molecular mechanism underlying observed pathological phenotypes in SNS, we generated drug-inducible KRAS pathogenic mutation expressing human NPC line using destabilization domain. In this cell line, several pathological phenotypes were rescued by ceasing the expression of the pathogenic mutation at the neuron stage. Stage-specific modulation of the KRAS variant will allow us to characterize molecular and cellular phenotypes that can be reversed. Taken together, these results indicate how KRAS p.G12V interrupts normal brain development and leads to neuropathology including epilepsy in SNS and suggest the possibility of relieving neurological symptoms in SNS patients by recently developed KRAS inhibitors.2

    Developmental Mechanisms of Epilepsy in Sebaceous Nevus Syndrome Caused by KRAS Mutation

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    Sebaceous nevus syndrome (SNS) is caused by somatic gain-of-function mutation of HRAS or KRAS. To examine the pathogenesis of SNS in brain, we generated the disease mouse model by overexpression of KRAS p.G12V in developing cortex using in utero electroporation. This model recapitulated histological manifestations observed in the patient. It showed abnormal neuronal differentiation, migration and neuronal morphogenesis. We investigated neuronal excitability at a cellular level, using patch clamp. Both neurons with and without KRAS p.G12V overexpression showed hyperexcitability. To elucidate the molecular mechanism underlying observed pathological phenotypes, we generated drug-inducible KRAS p.G12V expressing human NPC line using destabilization domain. In this cell line, several pathological phenotypes were rescued by ceasing the expression of KRAS p.G12V at the neuron. Stage-specific modulation of the KRAS variant will allow us to characterize molecular and cellular phenotypes that can be reversed. Taken together, these results indicate pathogenic mechanisms of SNS and suggest the possibility of relieving neurological symptoms in SNS patients by recently developed KRAS inhibitors1

    Dysregulation of RAS/MAPK Pathway During Neurodevelopment Leads to Severe Neuronal Migration Defects and Epileptogenesis

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    Sebaceous nevus syndrome (SNS) is a neuro-cutaneous disorder that shows neurological symptoms such as cerebral defects, intellectual disability and epilepsy with the skin lesion called nevus sebaceous. Recently, it has been reported that SNS is caused by somatic mutation of KRAS and HRAS during development. However, the pathological mechanism is poorly understood. Here, we report the pathological role of dysregulation of KRAS in SNS. We generated SNS mouse model by overexpressing KRAS p.G12V during developing cortex using in utero electroporation. Dysregulation of KRAS during cortical development led to defects in neuronal differentiation, migration and morphogenesis. However, neuronal proliferation was not affected, despite the oncogenic function of KRAS p.G12V in the other tissues. We also characterized neuronal activity in the disease model using whole cell path clamp. We observed difference in neuronal activity between KRAS p.G12V overexpressing neurons and control neurons. These results demonstrate that dysregulation of KRAS may lead to focal cortical dysplasia and epileptogenesis in the SNS patient.1

    Increased excitation-inhibition balance of the cortex in the mouse model of the focal malformation of cortical development

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    A typical symptom of epilepsy, a type of neurological disorder, includes a seizure caused by the sudden and disorderly over-excitement of neurons. This symptom can be controlled by medication in ~80 % of patients but not in ~20% of the intractable cases due to the gap between the target of the anticonvulsants and the causes. In the current study, we explored the abnormality of the circuits in the mouse model of focal malformation of cortical development (FMCD), which shows the intractable seizure. We found that the FMCD mouse model shows an increased intrinsic excitability not only in the neurons with somatic mutation, but also in neighboring neurons throughout cortical layers. Pyramidal neurons in the cortex with FMCD showed increased excitability by increased input resistance and depolarized resting membrane potential. In addition, a significant increase in the HCN-dependent rebounding current was observed in the mutant neurons. Consistently, we observed a dramatic decrease in the rheobase and increase in the firing frequency in response to the same amount of current injection. Furthermore, we found that the frequency of spontaneous inhibitory postsynaptic current (sIPSC) was significantly reduced. The reduction in the frequency of sIPSC can be attributed to reduction in (1) the excitability, (2) the release probability, or the number of inhibitory neurons of the cortical circuit. To distinguish these possibilities, we are currently analyzing the pathological features of the inhibitory neurons in FMCD mouse model.1
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