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Studies on the In Vitro Development of Cloned Embryos by Somatic Cell Nuclear Transfer in Korean Native Goats (재래산양의 체세포 핵이식에 의한 복제수정란의 체외발달에 관한 연구)

  • Park H. S.;Kim T. S.;Jung S. Y.;Lee Y. H.;Jung J. Y.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.105-112
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    • 2005
  • The present study was conducted to examine some factors affecting in vitro development of oocytes from somatic cell nuclear transfer (SCNT) in Korean native goats. Recipient oocytes were surgically collected after superovulation by using CIDR and FSH, PMSG, hCG and estrous synchronization in Korean Native goats. For nuclear transfer, the fibroblasts from caprine ear cells and fetal fibroblasts were surgically harvested and were cultured in vitro until cell confluency in serum-starvation condition (TCM-199 + $0.5\%$ FBS) for 3 to 5 days. The zona pellucidae of matured oocytes were partially drilled by laser irradiation. A single somatic cell was individually transferred into each enucleated oocyte. The reconstructed oocytes were then electrically fused and activated. Activated NT embryos were cultured in mSOF medium supplemented with $0.8\%\;BSA\;6\~7\;day\;at\;39^{\circ}C,\;5\%\;CO_2,\;5\%\;O_2,\;90\%\;N_2$ in air. There were no significant difference in the number of embryos cleaved and 4-cell development between the fibroblast nuclei from mature ear cells and fetal cells, but the rate of 8-cell development was higher (P<0.05) in ear cells $(40.5\%)$ than in fetal cells $(55.5\%)$. However, the embryo development to morula or blastocyst was not significantly different between both the groups$(6.7\%\;vs\;16.0\%)$, respectively. The number of embryo cleaved $(79.0\%)$ were higher (P<0.05) in the oocytes activated with ionomycin+6-DMAP than in the oocytes activated electrically $(9.5\%)$. The development of fused embryos to morula or blastocyst was found $15.6\%$ in ionomycin+6-DMAP, but no morula or blastocysts were developed in electrical stimulation. The development rate of SCNT embryos to morula or blastocyst was love. (P<0.05) in SCNT embryos $(19.0\%\;vs\;0.0\%)$ than that in parthenotes $(66.1\%\;vs\;59.1\%)$. In the parthenotes, the cleavage rate and development to morula or blastocyst were significantly higher (P<0.05) as $86.8\%\;and\;50.0\%$ in ovulated oocytes than in follicular oocytes $(69.0\%\;vs\;23.6\%)$, respectively. These results suggest that some factors Including superovulation treatment, oocyte source, maturation of follicular oocytes, activation method and culture condition may affect in vitro developmental capability of embryos produced by somatic cell nuclear transfer in Korean Native goats, and the fusion rate be greatly low compared with other species.

Improvement of Energy Efficiency of Plants Factory by Arranging Air Circulation Fan and Air Flow Control Based on CFD (CFD 기반의 순환 팬 배치 및 유속조절에 의한 식물공장의 에너지 효율 향상)

  • Moon, Seung-Mi;Kwon, Sook-Youn;Lim, Jae-Hyun
    • Journal of Internet Computing and Services
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    • v.16 no.1
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    • pp.57-65
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    • 2015
  • As information technology fusion is accelerated, the researches to improve the quality and productivity of crops inside a plant factory actively progress. Advanced growth environment management technology that can provide thermal environment and air flow suited to the growth of crops and considering the characteristics inside a facility is necessary to maximize productivity inside a plant factory. Currently running plant factories are designed to rely on experience or personal judgment; hence, design and operation technology specific to plant factories are not established, inherently producing problems such as uneven crop production due to the deviation of temperature and air flow and additional increases in energy consumption after prolonged cultivation. The optimization process has to be set up in advance for the arrangement of air flow devices and operation technology using computational fluid dynamics (CFD) during the design stage of a facility for plant factories to resolve the problems. In this study, the optimum arrangement and air flow of air circulation fans were investigated to save energy while minimizing temperature deviation at each point inside a plant factory using CFD. The condition for simulation was categorized into a total of 12 types according to installation location, quantity, and air flow changes in air circulation fans. Also, the variables of boundary conditions for simulation were set in the same level. The analysis results for each case showed that an average temperature of 296.33K matching with a set temperature and average air flow velocity of 0.51m/s suiting plant growth were well-maintained under Case 4 condition wherein two sets of air circulation fans were installed at the upper part of plant cultivation beds. Further, control of air circulation fan set under Case D yielded the most excellent results from Case D-3 conditions wherein air velocity at the outlet was adjusted to 2.9m/s.

A Reliable Protocol for transfection of mature primary hippocampal neurons using a neuron-glia co-culture system (신경세포-신경교세포 공동배양을 이용한 성숙한 해마신경세포의 효율적인 형질전환 방법)

  • Lee, Hyun-Sook;Cho, Sun-Jung;Jung, Yong-Wook;Jin, Ing-Nyol;Moon, Il-Soo
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.198-203
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    • 2007
  • DNA transfection is a powerful tool for studying gene functions. The $Ca^{2+}$-phosphate precipitation remains one of the most popular and cost-effective transfection techniques. Mature neurons are more resistant to transfection than young ones and most other cell types, and easy to die if microenvironment changes. Here, we report a transfection protocol for mature neurons. The critical modifications are inclusion of glial cells in culture and careful control of $Ca^{2+}$-phosphate precipitation under microscope. Cerebral glial cells were grown until ${\sim}70-80%$ confluence in DMEM/10% horse serum, which was thereafter replaced with serum-free Neurobasal/Ara-C, and 319 hippocampal neurons were plated onto the glial layer Formation of fine $DNA/Ca^{2+}$-phosphate precipitates was induced using Clontech $CalPhos^{TM}$ Mammalian Transfection Kit, and the size ($0.5-1\;{\mu}m$ in diameter) and density(about 10 particles/$100\;{\mu}m^2$) were carefully controlled by the time of incubation in the medium. This modified protocol can be reliably applied for transfection of mature neurons that are maintained longer than two weeks in vitro, resulting in 10-15 healthy transfected neurons per a well of 24-well plates. The efficacy of the protocol was verified by punctate expression of $pEGFP-CaMKII{\alpha}$, a synaptic protein, and diffuse expression of pDsRed2. Our protocol provides a reliable method for transfection of mature neurons in vitro.

Expression and Purification of the Phosphatase-like Domain of a Voltage-Sensing Phosphatase, Ci-VSP (막 전위 감지 탈인산화 효소, Ci-VSP의 유사 탈인산화 효소 도메인의 발현과 정제)

  • Kim, Sung-Jae;Kim, Hae-Min;Choi, Hoon;Kim, Young-Jun
    • Journal of Life Science
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    • v.21 no.7
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    • pp.1032-1038
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    • 2011
  • Recently identified Ciona intestinalis voltage sensor-containing phosphatase (Ci-VSP) consists of an ion channel-like transmembrane domain (VSD) and a phosphatase-like domain. Ci-VSP senses the change of membrane potential by its VSD and works as a phosphoinositide phosphatase by its phosphatase domain. In this study, we present the construction of His-tagged phosphatase-like domain of Ci-VSP, its recombinant expression and purification, and its enzymatic activity behavior in order to examine the biochemical behavior of phosphatase domain of Ci-VSP without interference. We found that Ci-VSP(248-576)-His can be eluted with an elution buffer containing 25 mM NaCl and 100 mM imidazole during His-tag purification. In addition, we found the proper measurement condition for kinetics study of Ci-VSP(248-576)-His against p-nitrophenyl phosphate (pNPP). We measured the kinetic constant of Ci-VSP(248-576)-His at $37^{\circ}C$, pH 5.0 or 5.5, under 30 min of reaction time, and less than $2.0\;{\mu}g$ of protein amount. With these conditions, we acquired that Ci-VSP(248-576)-His has $K_m$ of $354{\pm}0.143\;{\mu}M$, $V_{max}$ of $0.0607{\pm}0.0137\;{\mu}mol$/min/mg and $k_{cat}$ of $0.359{\pm}0.009751\;min^{-1}$ for pNPP dephosphorylation. Therefore, we produced a pure form of Ci-VSP(248-576)-His, and this showed a higher activity against pNPP. This purified protein will provide the road to a structural investigation on an interesting protein, Ci-VSP.

THE EFFECT OF FGF-MEDIATED FGFR SIGNALING ON THE EARLY MORPHOGENESIS AND MAINTENANCE OF THE CRANIAL SUTURE (FGF-mediated FGFR signaling이 두개봉합부의 초기형태발생 및 유지기전에 미치는 영향)

  • Sue, Kyung-Hwan;Park, Mi-Hyun;Ryoo, Hyun-Mo;Nam, Soon-Hyeun;Kim, Young-Jin;Kim, Hyun-Jung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.26 no.4
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    • pp.652-663
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    • 1999
  • Craniosynostosis, the premature fusion of cranial sutures, presumably involves disturbance of the interactions between different tissues within the cranial sutures. Interestingly, point mutaions in the genes encoding for the fibroblast growth factor receptors(FGFRs), especially FGFR2, cause various types of human craniosynostosis syndromes. To elucidate the function of these genes in the early morphogenesis of mouse cranial sutures, we first analyzed by in situ hybridization the expression of FGFR2(BEK) and osteopontin, an early marker of osteogenic differentiation, in the sagittal suture of calvaria during embryonic(E15-E18) and postnatal stage(P1-P3). FGFR2(BEK) was intensely expressed in the osteogenic fronts, whose cells undergo differentiation into osteoprogenitor cells that ultimately lay down the bone matrix. Osteopontin was expressed throughout the parietal bones excluding the osteogenic fronts, the periphery of the parietal bones. To further examine the role of FGF-mediated FGFR signaling in cranial suture, we did in vitro experiments in E15.5 mouse calvarial explants. Interestingly, implantation of FGF2 soaked beads onto both the osteogenic fronts and mid-mesenchyme of sagittal suture after 36 hours organ culture resulted in the increase of the tissue thickness and cell number around FGF2 beads, moreover FGF4-soaked beads implanted onto the osteogenic fronts stimulated suture closure due to an accelerated bone growth, compared to FGF4 beads placed onto mid-mesenchyme of sagittal suture and BSA control beads. In addition FGF2 induced the ectopic expression of osteopontin and Msx1 genes. Taken together, these data indicate that FGF-mediated FGFR signaling has a important role in regulating the cranial bone growth and maintenance of cranial suture, and suggest that FGF-mediated FGFR signaling is involved in regulating the balance between the cell proliferation and differentiation through inducing the expression of osteopontin and Msx1 genes.

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A Measures to Implements the Conservation and Management of Traditional Landscape Architecture using Aerial Photogrammetry and 3D Scanning (전통조경 보존·관리를 위한 3차원 공간정보 적용방안)

  • Kim, Jae-Ung
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.38 no.1
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    • pp.77-84
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    • 2020
  • This study is apply 3D spatial information per traditional landscape space by comparing spatial information data created using a small drone and 3D scanner used for 3D spatial information construction for efficient preservation and management of traditional landscaping space composed of areas such as scenic sites and traditional landscape architectures. The analysis results are as follows. First, aerial photogrammetry data is less accurate than 3D scanners, but it was confirmed to be more suitable for monitoring landscape changes by reading RGB images than 3D scanners by texture mapping using digital data in constructing orthographic image data. Second, the orthographic image data constructed by aerial photogrammetry in a traditional landscaping space consisting of a fixed area, such as Gwanghalluwon Garden, produced visually accurate and precise results. However, as a result of the data extraction, data for trees, which is one of the elements that make up the traditional landscaping, was not extracted, so it was determined that 3D scanning and aerial surveying had to be performed in parallel, especially in areas where trees were densely populated. Third, The surrounding trees in Soswaewon Garden caused many errors in 3D spatial information data including topographic data. It was analyzed that it is preferable to use 3D scanning technology for precise measurement rather than aerial photogrammetry because buildings, landscaping facilities and trees are dense in a relatively small space. When 3D spatial information construction data for a traditional landscaping space composed of area using a small drone and a 3D scanner free from temporal and spatial constraints and compared the data was compared, the aerial photogrammetry is effective for large site such as Hahoe Village, Gyeongju and construction of a 3D space using a 3D scanner is effective for traditional garden such as Soswaewon Garden.

Treatment Margin Assessment using Mega-Voltage Computed Tomography of a Tomotherapy Unit in the Radiotherapy of a Liver Tumor (간종양 방사선치료 시 토모테라피 메가볼트 CT를 이용한 치료 여백 평가)

  • You, Sei-Hwan;Seong, Jin-Sil;Lee, Ik-Jae;Koom, Woong-Sub;Jeon, Byeong-Chul
    • Radiation Oncology Journal
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    • v.26 no.4
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    • pp.280-288
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    • 2008
  • Purpose: To identify the inter-fractional shift pattern and to assess an adequate treatment margin in the radiotherapy of a liver tumor using mega-voltage computed tomography (MVCT) of a tomotherapy unit. Materials and Methods: Twenty-six patients were treated for liver tumors by tomotherapy from April 2006 to August 2007. The MVCT images of each patient were analyzed from the $1^{st}$ to the $10^{th}$ fraction for the assessment of the daily liver shift by four groups based on Couinard's proposal. Daily setup errors were corrected by bony landmarks as a prerequisite. Subsequently, the anterior-, posterior-, right-, and left shifts of the liver edges were measured by maximum linear discrepancies between the kilo-voltage computed tomography (KVCT) image and MVCT image. All data were set in the 2-dimensional right angle coordinate system of the transverse section of each patient's body. Results: The liver boundary shift had different patterns for each group. In group II (segment 2, 3, and 4), the anterior mean shift was $2.80{\pm}1.73\;mm$ outwards, while the left mean shift was $2.23{\pm}1.37\;mm$ inwards. In group IV (segment 7 and 8), the anterior-, posterior-, right-, and left mean shifts were $0.15{\pm}3.93\;mm$ inwards, $3.15{\pm}6.58\;mm$ inwards, $0.60{\pm}3.58\;mm$ inwards, and $4.50{\pm}5.35\;mm$ inwards, respectively. The reduced volume in group II after MVCT reassessment might be a consequence of stomach toxicity. Conclusion: Inter-fractional liver shifts of each group based on Couinard's proposal were somewhat systematic despite certain variations observed in each patient. The geometrical deformation of the liver by respiratory movement can cause shrinkage in the left margins of liver. We recommend a more sophisticated approach in free-breathing mode when irradiating the left lobe of liver in order to avoid stomach toxicity.

피부 병변은 닭 마렉병 진단에 결정적 단서를 제공한다

  • 조경오;박남용
    • Korean Journal of Veterinary Pathology
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    • v.6 no.2
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    • pp.101-104
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    • 2002
  • 닭의 전신장기에 림프종 발생이 특정인 마렉병(Marek's disease; MD)은 림프구성 백혈병(Lymphoid leukosis; LL)과 병리학적 소견이 매우 유사하여 감별이 요구된다. 마렉병 바이러스(Marek's disease virus; MDV)는 질병초기에서 후기에 이르기까지 모낭상피세포에 세포용해성 감염을 지속적으로 일으킨다. 세포용해성 감염이 있는 모낭상피세포는 변성내지 괴사되어 있고 핵내봉입체가 관찰된다. 또한 세포용해성 감염이 있는 모낭상피세포 바로 밑의 진피와 깃털 수질(feather pulp)내의 혈관주위에 림프구 침윤이 관찰된다. 이러한 피부병변은 MD의 특징적인 병변들로서 LL과 감별할 수 있는 결정적인 단서이다. 본 고에서는 MD에 관한 전반적인 것과 특히 MD 진단을 위한 피부병변의 유용성에 대해서 자세히 논하고자 한다. 양계산업에 문제를 야기하고 있는 림프구 증식성 종양은 크게 마렉병(Marek’s disease; MD)과 백혈병(Lymphoid leukosis; LL)이 있다. 이들 질환의 원인체들이 분리되기 전까지는 이들 질병들은 발병부위에 따라 질병명을 붙였다. 즉, 내부장기냐 근육에 발생한 것은 visceral lymphomatosis, 피부에 발생한 것은 skin leukosis, 말초신경의 병변은 poly­n neuritis, neuritis, neurolymphomatosis gallinarum, range paralyis로 불리었다. 또한 눈에 발생한 것은 blindness, gray eye, iritis, uveitis, ocular lymphomatosis라고 불리었다. 1961년에 Biggs는 이러한 림프구 증식성 질병을 마렉병과 백혈병 으로 분류하자는 제안을 하였고, 드디어 1960년대 중반에 림프구 증식성 병변 중의 일부에서 herpesvirus가 분리되어서 Biggs가 제안한 병명인 마렉병을 본격적으로 사용하게 되었다. MD는 마렉병 바이러스(Marek’s disease virus; MDV)가 원 인제로서 닭에 전염성이 강한 염증성에서 종양성의 병변을 내부장기, 피부, 근육, 안구, 중추신경계, 말초신경계 등에 일 으킨다. MDV는 림프종을 유발하기 때문에 처음에는 사람에서 림프종을 유발하는 Epstein-Bar 바이러스와 관련이 있을 것으로 생각되어 gamma-herpesvirus로 분류되었지만, MDV의 게놈 구조와 세포배양에서 빠르게 성장한다는 점 때문에 지금은 alpha-herpesvirus로 재분류 되었다. MDV는 바이러스 중화시험과 한천 겔 침강법에 의해서 3개의 혈청형으로 분류 된다. 혈청형 1은 종양원성 바이러스와 종양원성 바이러스의 계대배양에 의한 약독주가 있다. 혈청형 2는 자연적으로 발 생하는 비종양원성 닭의 MDV이고, 혈정형 3은 바종양원성 칠면조 herpesvirus (HVT)이다. 림프종을 유발하는 MDV 감염은 4개의 과정, 즉 초기 세포용해성 감염, 잠복감염, 후기 세포용해성 감염, 마지막으로 종양화로 나눌 수 있다. 감염의 경로를 보면, 흡입된 MDV는 폐의 대식세포에 감염한 후 전선 장기로 MDV를 전파 시킨다. 특히, 흉선, F냥, 비장 등의 림프구에 초기 세포용해성 감염을 일으키는데, B 림프구가 주로 감염된다. 세포용해성 감염음 방어하기 위해 몰려든 T 림프구가 활성화가 되면, T 세포도 감염되게 된다. 잠복감염은 여러 가지 사이토카인 등 을 포함한 면역반응에 의해서 일어나며, 이 때 잠복감염된 세포는 특히 혈중의 T 세포라고 한다. 혈중 MDV 감염세포 는 피부 모낭상피세포, 선장 등 상피세포 유래의 조직에 MDV를 전파 시켜서 이들 조직에서 후기 세포용해성 감염이 일어나게 한다. 후기 세포용해성 감염을 유발하는 것은 이러한 MDV가 감염된 혈중 림프구라는 증거는 혈중의 세포 외에는 감염성 바이러스가 없기 때문이다. 후기 세포용해성 감염이 있는 후 육안적 혹은 현미경적으로 검출이 가능한 림프종이 여러 장기에서 관찰된다. MDV가 감염되면 병변 발생부위에 따라서 다양한 임상증상이 발생한다. 즉 내부장기에 병변이 있을 경우는 침울, 체중감소, 산란율 저하 등이 발생하며, 신경계는 발생부위별 신경증상이 일어난다. 이러한 장기나 조직에서는 육안적으로 백색에서 회백색의 종양성 병변이 관찰된다. 말초신경에 병변이 발생한 경우에는 특히 좌골신경 및 신경총에서 호발하는데, 이들 조직은 편측성 혹은 양측성으로 종창되어 있다. 안구는 간혹 육안적으로 식별이 가능한 홍채 퇴색 및 증식 병변이 관찰된다. 피부형 MD는 특히 육용계에서 문제가 되고 있으며, 산란계의 내장형 MD가 발생한 경우에도 피부를 자세히 설펴보면 피부형 MD를 간혹 관찰할 수 있다. 피부형 MD는 모낭주위에 원형의 결절형태로 발생하는데, 이들 병변이 커지면 바로 옆의 병변과 융합 되어 큰 결절을 형성하기도 한다. (중략)

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Study on Features of Software Cyborg in the Virtual Game -PS4 ocusing Game- (가상게임에 나타나는 소프트웨어 사이보그특징에 대한 고찰 -PS4 <언틸던> 게임을 중심으로-)

  • Kim, Dae-Woo
    • Cartoon and Animation Studies
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    • s.41
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    • pp.279-306
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    • 2015
  • This paper is a study of the changing nature of software for virtual Cyborg self and the virtual body that occur in the game from a philosophical point of view. Looking broadly, the cyborg concept refers to the combination of man and machine. Specifically, there is a hardware cyborg organism to combine human and restoration of machine In addition, there is software cyborg by electronic the human brain of converting a virtual body. Virtual games are cases software-Cyborg applied. In the game , There seems to have characteristics of virtual body and ego that different from general cyborg meaning. To analyze the features, I applied the concept of software-cyborg of Hans morabek and the multiple selves in cyberspace properties of Kim Sun-Hee. generally, software cyborg cloning the brain type tended to invalidate the body due to the nature of the virtual world. But If you look at third-person's view and the game character that made from real actors, it is pursuing the realism of photographic images and it stressed the need for a virtual body in order to maintain the psychological identity of the player. And, The game player crosses the eight characters to choose while completing the mission. This is a big role in the reality ego leads to the desired final ending with the selection and experience to be experienced as self-replication to multiple. These cyber multi-ego looks for an active and positive features compared to the multi-ego in the real world and highlights the advantages of the software cyborg. Game The characteristics of the final result varies depending on the selection of the player. The life and death of a friend is determined by the relationship between the characters friendship. In this case, the virtual self is empirically through trial and error, moral, and try to select the desired setting the standard for intuitive and self own choice. Also It can be fused to the knowledge of multiple selves as one step is formed by a high spiritual introspection. This process is a positive interpretation of the world and their own forms of mental reflection through self-overcoming human, Nietzsche is said that the process is Wibeomenswi.

THE CHANGE OF THE CONFIGURATION OF HYDROXYAPATITE CRYSTALS IN ENAMEL BY CHANGES OF PH AND DEGREE OF SATURATION OF LACTIC ACID BUFFER SOLUTION (유산 완충용액의 pH 및 포화도 변화에 따른 법랑질 내 수산화인회석 결정 형태의 변화)

  • Chon, Young-Eui;Jung, Il-Young;Roh, Bung-Duk;Lee, Chan-Young
    • Restorative Dentistry and Endodontics
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    • v.32 no.6
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    • pp.498-513
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    • 2007
  • Since it was reported that incipient enamel caries can be recovered, previous studies have quantitatively evaluated that enamel artificial caries have been, remineralized with fluoride showing simultaneously the increase of width of surface layer and the decrease of width of the body of legion. There is, however, little report which showed that remineralization could occur without fluoride. In addition, the observations on the change of hydroxyapatite crystals also have been scarcely seen. In this study, enamel caries in intact premolars or molars was induced by using lactic acidulated buffering solutions over 2 days. Then decalcified specimens were remineralized by seven groups of solutions using different degree of saturation(0.212, 0.239, 0.301, 0.355) and different pH(5.0, 5.5, 6.0) over 10 days. A qualitative comparison to changes of hydroxyapatite crystals after fracturing teeth was made under SEM(scanning electron microscopy) and AFM(atomic force microscopy). The results were as follows: 1. The size of hydroxyapatite crystals in demineralized area was smaller than the normal ones. While the space among crystals was expanded, it was observed that crystals are arranged irregularly. 2. In remineralized enamel area, the enlarged crystals with various shape were observed when the crystals were fused and new small crystals in intercrystalline spaces were deposited. 3. Group 3 and 4 with higher degree of saturation at same pH showed the formation of large clusters by aggregation of small crystals from the surface layer to the lesion body than group 1 and 2 with relatively low degree of saturation at same pH did. Especially group 4 showed complete remineralization to the body of lesions. Group 5 and 6 with lower pH at similar degree of saturation showed remineralization to the body of lesions while group 7 didn't show it. Unlike in Group 3 and 4, Group 5 and 6 showed that each particle was densely distributed with clear appearance rather than crystals form clusters together.