• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.22-22
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• 2001

To produce reconstituted rabbit embryos with fetal fibroblasts, the present study was evaluated the efficiencies of the different fusion and activation conditions as assessments of subsequent development and chromosome in the embryos. New Zealand White rabbits were used throughout the study. Fetal fibroblasts collected from 22-d of fetuses were cultured in DMEM ＋ 10% FBS in 5% $CO_2$ in air. The culture was maintained for 10 passages. In every passage half of cell suspension were kept In frozen. From rabbits treated with FSH in 30% PVP solution and hCG, oocytes were surgically collected from oviducts at 14 h post-hCG injection and stripped off their cumulus cells by re-pipetting in a 300 IU hyaluronidase solution. Oocytes with an extruded first polar body and dense cytoplasm were enucleated by micromanipulation in Ham's F-10 medium＋7.5 g/$m\ell$ cytochalasin B. Euncleation was confirmed under a fluorescence microscope after staining with 5 g/$m\ell$ bisbenzimide for 2 min. Each enucleated oocyte was injected with a fetal fibroblast into a perivitelline space. Reconstructed eggs were compared fusion rates either at 2.0 ㎸/cm or 1.6 ㎸/cm(60 sec, double pulses). After fusion, all eggs were activated with the combination of 5 M ionomycin (5 min) and 10 g/$m\ell$ cycloheximide (CHX, 3h), and cultured in CRlaa medium and transferred into TCM199＋10% FBS on day 3. Although there was not significantly differ in fusion rate between treatments (60%, 2.0 ㎸/cm vs. 79.4%, 1.6 ㎸/cm), none of them in the eggs fused with 2.0 ㎸/cm developed to blastocyst. In comparison of development and chromosome status between different activation treatments (Group 1; 5 M ionomycin/10 g/$m\ell$ CHX, Group 2; 5 M ionomycin/5 g/$m\ell$ CHX ＋ 2 mM DMAP after fusion with 1.6 ㎸/cm), there were not differ in cleavage and development rates (67.3% and 28.9% in Group 1; 67% and 33% in Group 2). All out of 8 embryos evaluated in Group 1 appeared a normal diploid chromosome sets and mean number of cells (Mean SEM) on day 4.5 of culture was 141.5 23.15 (n=8). It can be concluded that the use of cycloheximide has not happened in chromosome abnormalities, and fetal fibroblasts can be used for cloning in rabbit.

• Geomechanics and Engineering
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• v.8 no.1
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• pp.81-95
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• 2015

The permeability behavior of Ariake clays has been investigated by constant rate of strain (CRS) consolidation tests with vertical or radial drainage. Three types of Ariake clays, namely undisturbed Ariake clay samples from the Saga plain, Japan (aged Ariake clay), clay deposit in shallow seabed of the Ariake Sea (young Ariake clay) and reconstituted Ariake clay samples using the soil sampled from the Saga plain, were tested. The test results indicate that the deduced permeability in the horizontal direction ($k_h$) is generally larger than that in the vertical direction ($k_v$). Under odometer condition, the permeability ratio ($k_h/k_v$) increases with the vertical strain. It is also found that the development of the permeability anisotropy is influenced by the inter-particle bonds and clay content of the sample. The aged Ariake clay has stronger initial inter-particle bonds than the young and reconstituted Ariake clays, resulting in slower increase of $k_h/k_v$ with the vertical strain. The young Ariake clay has higher clay content than the reconstituted Ariake clay, resulting in higher values of $k_h/k_v$. The microstructure of the samples before and after the consolidation test has been examined qualitatively by scanning electron microscopy (SEM) image and semi-quantitatively by mercury intrusion porosimetry (MIP) tests. The SEM images indicate that there are more cut edges of platy clay particles on a vertical plane (with respect to the deposition direction) and there are more faces of platy clay particles on a horizontal plane. This tendency increases with the increase of one-dimensional (1D) deformation. MIP test results show that using a sample with a larger vertical surface area has a larger cumulative intruded pore volume, i.e., mercury can be intruded into the sample more easily from the horizontal direction (vertical plane) under the same pressure. Therefore, the permeability anisotropy of Ariake clays is the result of the anisotropic microstructure of the clay samples.

• The Korean Journal of Mycology
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• v.24 no.2 s.77
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• pp.111-126
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• 1996

Transfer of the isolated nuclei from Lentinus edodes into protoplasts of Pleurotus florida was induced with polyethlene glycol (PEG) and $CaCl_2$. The intergeneric transfer products were classified into nuclear hybrid, heterokaryon or synkaryon, and reconstituted cell. These progenies except nuclear hybrids formed mature fruiting bodies on sawdust rice bran medium. Formation of fruit bodies was influenced by several factors such as light, temperature, nutrition and physic state of the culture media. Most of fruiting body characters were similar to those of P. florida in synkaryon and L. edodes in reconstituted cell, respectively. All these basidiocarps had clamp connections though initial heterokaryon colonies were lacking. Isozyme patterns of intergeneric progenies were quite different from those of parents. DNA polymorphisms of transfer products were also compared by random amplified polymorphic DNAs (RAPD) analysis based on polymerase chain reaction. The RAPD patterns were different from those of donor and recipient. DNA fingerprints ranged in size from 0.25 to 4.0 Kb. On the basis of RAPD, the transfer products were classified into five groups. Two synkaryon were analysed with distribution of progenies and segregation of genetic markers by random spore analyses. The genetic markers were segregated into wild type and riboflavine requiring auxotrophs.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.129-142
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• 2002

Epithelial-mesenchymal interaction plays a important role in cell growth and differentiation. This interaction is already well known to have an importance during the organ development as well as cell growth and differentiation. However, in vitro experimental model is not well developed to reproduce in vivo cellular microenvironment which provide a epithelial-mesenchymal interaction. Because conventional monolayer culture lacks epithelial-mensenchymal interaction, cultivated cells have an morphologic, biochemical, and functional characteristics differ from in vivo tissue. Moreover, it's condition is not able to induce cellular differention due to submerged culture condition. Therefore, the aims of this study were to develop and evaualte the in vitro experimental model that maintains epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally reconstituted oral keratinocytes by histological and immunohistochemical analysis. The results were as follow; 1. Gingival keratinocytes reconstituted by three-dimensional organotypic culture revealed similar morphologic characteristics to biopsied patient specimen showing stratification, hyperkeratinosis, matutation of epithelial architecture. 2. Connective tissue structure was matured, and there is no difference during stratification period of epithelial 3-dimensional culture. 3. The longer of air-exposure culture on three-dimensionally reconstituted cells, the more epithelial maturation, increased epithelial thickness and surface keratinization 4. In reconstitued mucosa, the whole epidermis was positively stained by anti-involucrin antibody, and there is no difference according to air-exposured culture period. 5. The Hsp was expressed in the epithelial layer of three-dimensionally cultured cells, especially basal layer of epidermis. The change of Hsp expression was not significant by culture stratification. 6. Connexin 43, marker of cell-cell communication was revealed mild immunodeposition in reconstitued epithelium, and there is no significant expression change during stratification. These results suggest that three-dimensional oragnotypic co-culture of normal gingival keratinocytes with dermal equivalent consisting type I collagen and gingival fibroblasts results in similar morphologic and immunohistochemical characteristics to in vivo patient specimens. And this culture system seems to provide adequate micro-environment for in vitro tissue reconstitution. Therefore, further study will be focused to study of in vitro gingivitis model, development of novel perioodntal disease therapeutics and epithelial-mensenchymal interaction.

• Journal of the Korean Geotechnical Society
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• v.22 no.6
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• pp.41-49
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• 2006

This paper deals with development and dissipation of excess pore pressure induced by the cyclic load. Cyclic triaxial tests on reconstituted samples of weathered residual soils, which were widely used as construction materials in Korea, were performed. Test results showed that excess pore pressures developed under undrained condition increased with the increase of cyclic loads and confining pressures. And a new concept based on modified excess pore pressure ratio (MEPPR) was proposed for simply estimating excess pore pressures in terms of the number of cyclic load, irrespective of cyclic loads and confining pressures. Also, it was proposed that excess pore pressure ratio (EPPR) could be effectively utilized to estimate volumetric strains during dissipation which decreased as confining pressures increased. Consequently, concept and method to effectively estimate settlements under non-liquefied condition, induced by dynamic loads such as earthquake loads were evaluated based on laboratory test results for reconstituted weathered residual soils.

• Korean Journal of Animal Reproduction
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• v.23 no.3
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• pp.271-278
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• 1999

This study was conducted to investigate the cytogenetic properties, in vitro development, and their relationship in the bovine reconstituted embryos following cell cycle-controlled nuclear transfer. Sixteen-cell stage embryos were treated by nocodazole, and after release from nocodazole treatment, their blastomeres were separated and allowed to subsequent cleavage. Blastomeres within 1.5 h post cleavage(hpc) and at 3hpc were transferred to enucleated oocytes at MII-phase or S-phase. Donor nuclei transferred into M II-phase recipients underwent various nuclear remodeling, such as extrusion of a polar body(PB)-like structure, premature chromosome condensation(PCC) and chromatin modifications. These nuclear remodeling patterns varied by the time post cleavage of donor blastomeres. Developmental rate to the blastocyst stage differed with time post cleavage of donor blastomeres and existence of a PB-like structure. Whereas do-nor nuclei transferred into S-phase oocytes did not undergo PCC and other major modifications, and their developmental potentials less depended on the nuclei types. This result confirms that the nuclear remodeling type differs with donor and recipient cell cycle stage, which affect the development of reconstituted bovine embryos.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.149.3-150
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• 2003

CYP3A4 is the most abundant human CYP and oxidizes a diversity of substrates. including various drugs. steroids. and carcinogens. A variety of metal ions are known to affect microsomal monooxygenase activities. Effects of a series of divalent metal ions on the CYP3A4-catalyzed reaction of reconstituted system containing purified CYP3A4. NADPH-P450 reductase (NPR), and cytochrome b5 (b5) were examined. (omitted)

• Proceedings of the Korean Biophysical Society Conference
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• 1996.07a
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• pp.6-6
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• 1996

Gap junction channels were reconstituted into unilamellar liposomes using immunoaffinity purified connexin 32 gap junction protein from rat liver. Vesicles containing open channels and close channels were separated by means of iso-osmolar sucros density gradient sedimentation. The open channels formed in lipid vesicles were permeable to a fluorescent dye molecule, lucifer yellow of which the hydrodynamic size is similar to pore size of gap junctions in vivo. (omitted)

• Proceedings of the Korean Biophysical Society Conference
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• 1996.07a
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• pp.35-35
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• 1996

Nitric oxide (NO) has been reported to have many roles in vivo ranging from the neurotransmitter in brain to the relaxant in smooth muscles. Recently, using inside-out patches, Bolotina et al. (1) showed that relaxing effect of NO is aortic smooth muscle is through direct activation of Ca2+-activated K+ channels (maxi-K), resulting in hyperpolarization. (omitted)

• Journal of Dairy Science and Biotechnology
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• v.14 no.2
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• pp.185-193
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• 1996

This study was to investigate principle of freeze concentration such as nucleation, crystal growth and procedure of freeze concentration, essential factor and application of freeze concentration. Especially, quality of milk was emphasized. For exemple, in sensory evaluation of freeze concentrated and reconstituted skim milk and whole milk, taste, color, mouth feel and texture were superior to control. Recently developed technique of freeze concentration for quality milk and dairy products may be expected for advanced quality of various milk and dairy products in near future.