• 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.

• Journal of Life Science
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• v.25 no.5
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• pp.594-600
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• 2015

Protein-protein interactions have a critical role in the regulation of many cellular functions. Postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domain is one of domains that mediate protein-protein interactions. PDZ domains typically bind to the specific motif at the carboxyl (C)-terminal end of partner proteins. Multi-PDZ domain protein 1 (MUPP1), which has 13 PDZ domains, serves a scaffolding function for structure proteins and signaling proteins, but the cellular function of MUPP1 has not been fully elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and muskelin. Muskelin was recently identified as a GABA_A receptor (GABA_AR) α1 subunit binding protein and known to have a role in receptor endocytosis and degradation. Muskelin bound to the 3^rd PDZ domain, but not to other PDZ domains of MUPP1. The C-terminal end of muskelin was essential for the interaction with MUPP1 in the yeast two-hybrid assay. When co-expressed in HEK-293T cells, muskelin but not the C-terminal deleted muskelin was co-immunoprecipitated with MUPP1. In addition, MUPP1 co-localized with muskelin at the same subcellular region in cells. These findings collectively suggest that MUPP1 or its interacting proteins could modulate GABA_AR trafficking and turnover through the interaction with muskelin.

• Journal of Life Science
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• v.27 no.10
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• pp.1111-1120
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• 2017

This study was conducted to select pepper lines that were tolerant to excessive water injury among the pepper germplasm and investigate the genetic characteristics of those lines to contribute to the breeding of pepper cultivars with stable productivity in abnormal weather. Each of the tolerant and susceptible lines went through immersion treatment, and differentially expressed genes between them were analyzed. The tolerant line showed increased expression of the CA02g26670 gene, which is involved in the CONSTANS protein pathway and regulation of flowering by day length, but it exhibited decreased expressions of CA01g21450, CA01g22480, CA01g34470, CA02g00370 and CA02g00380. The susceptible line showed increased gene expressions of CA02g09720, CA02g21290, CA03g16520, CA07g 02110, and CA12g17910, which are involved in the inhibition of proteolytic enzyme activity, DNA binding, inhibition of cell wall-degrading enzyme, and inhibition of nodulin, respectively. Meanwhile the expressions of CA02g02820, CA03g21390, CA06g17700 and CA07g18230 decreased in the susceptible line, in relation to calcium-ion binding, high temperature, synthesis of phosphocholine and cold stress, respectively. The expressions of genes related to apoptosis and peroxidase increased, while that of CA02g16990, which functions as a nucleoside transporter, decreased in both the tolerant and susceptible lines. Based on the different gene expressions between the tolerant and susceptible lines, further studies are needed on breeding abiotic stress-tolerant lines.

• Journal of Life Science
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• v.27 no.10
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• pp.1191-1198
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• 2017

Vesicles and organelles are transported along microtubule and delivered to appropriate compartments in cells. The intracellular transport process is mediated by molecular motor proteins, kinesin, and dynein. Kinesin is a plus-end-directed molecular motor protein that moves the various cargoes along microtubule tracks. Kinesin 1 is first isolated from squid axoplasm is a dimer of two heavy chains (KHCs, also called KIF5s), each of which is associated with the light chain (KLC). KIF5s interact with many different binding proteins through their carboxyl (C)-terminal tail region, but their binding proteins have yet to be specified. To identify the interacting proteins for KIF5A, we performed the yeast two-hybrid screening and found a specific interaction with Ras-GTPase-activating protein (GAP) Src homology3 (SH3)-domain-binding protein 2 (G3BP2), which is involved in stress granule formation and mRNA-protein (mRNP) localization. G3BP2 bound to the C-terminal 73 amino acids of KIF5A but did not interact with the KIF5B, nor the KIF5C in the yeast two-hybrid assay. The arginine-glycine-glycine (RGG)/Gly-rich region domain of G3BP2 is a minimal binding domain for interaction with KIF5A. However, G3BP1 did not interact with KIF5A. When co-expressed in HEK-293T cells, G3BP2 co-localized with KIF5A and was co-immunoprecipitated with KIF5A. These results indicate that G3BP2, which was originally identified as a Ras-GAP SH3 domain-binding protein, is a protein that interacts with KIF5A.

• Restorative Dentistry and Endodontics
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• v.8 no.1
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• pp.123-131
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• 1982

The purpose of this study was to observe the responses of the remaining pulp tissue after pulpotomy upon the several kinds of $Ca(OH)_2$ products and the responses of periapical tissue upon some root canal filling materials after extirpation. For pulpotomy, the class V cavities were prepared on the premolars, molars and upper canines, and the pulp was amputated. Each drug was placed over the amputated tissue and cavity was sealed with zinc oxide eugenol cement. The drugs which were used for the study were Dycal (Caulk Co. U.S.A.), Cavitec (Kerr Co. U.S.A.), Calvital, Nobudyne and Neodyne (Neo Dental Chemical Products). For extirpation, the endodontic cavities were prepared on the lingual surfaces of anterior teeth, and the pulp tissues were extirpated as routine method. After enlarging, irrigation, and measuring of root length by taking X-ray, each root canal filling material was filled in the canal with gutta percha cone, and endodontic cavity was sealed with zinc oxide eugenol cement. Zinc oxide eugenol, $Ca(OH)_2$ (Eli Lilly Co. U.S.A.) and Vitapex (Neo Dental Chemical Products) were used as root canal filling materials. Animals were sacrificed after 1, 3 and 6 weeks following the operation. The teeth were decalcified in formic acid, sectioned and stained with hematoxylin eosin. Microscopic examination revealed as follows. 1. Dycal: The dentin bridge formation was observed at the 3rd week after pulpotomy. Inflammatory conditions which were infiltration of inflammatory cells and dilatation of blood vessels were kept in remaining pulp tissue at the 6th week. 2. Calvital: The dentin bridge was observed at the 1st week after pulpotomy. As the time clasped, the pulp tended to be the fibrous degeneration. 3. Cavitec, Nobudyne and Neodyne: In the case of Cavitec and Nobudyne, the incompleted and irregular dentin bridge was observed at the 6th week, and in Neodyne, was observed at the 3rd week. The severe inflammatory changes were seen in the remaining pulp tissue. As the time clasped, the fibrous degeneration tended to spread in the remaining pulp tissue. 4. $Ca(OH)_2$: Osteocementum was formed at the 3rd week, the matrix of cementum and dentin were resorted, and infiltration of lymphocytes was seen in periapical tissue when $Ca(OH)_2$ was used as canal-filling materials. S. ZOE and Vitapex The cementum like substance was seen in periapical portion at the 1st week, when ZOE and Vitapex were used as root canal filling materials. As the time elapsed, the matrix of cementum and dentin tended to be resorted. At the 6th week, the inflammatory condition of periapical tissue was continued in the case of ZOE, but was reduced in the case of Vitapex.

• Journal of Gifted/Talented Education
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• v.14 no.2
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• pp.1-18
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• 2004

This study attempted to survey the present status of the eight students who, 17 years from now, had participated in a special program for 170 days preparing for early entrance to the KAIST. With six students who responded to the questionnaire previously sent, the study focused on each participant's achievement at present, his evaluation on Accelerated Education, his personal opinions on early entrance to university and Gifted Science Education. From the survey, we could find out that three participants had received a doctorate and they are currently working as researchers at research center or professors at university. The other three participants are working as researchers at research center after receiving the M. S. Academic achievements vary according to each participant, but three of them, who ranked high in research achievement, had their articles published in international journals of the SCI level even when they were between 25 and 28 of age. Two of them in particular showed outstanding engineering and developing the commercial modem for IMT 2000 for the first time in Korea. One participant entered Harvard University to major in physics and afterwards studied the law in UCLA. He is now a professor in the law department. Concerning the evaluations on Accelerated Education and early entrance system, four participants answered positively while two participants expressed negative attitudes. When asked their opinions on the Gifted Education in Korea, such problems were pointed out as too much pressure of entering the first-class universities, the tendency to avoid choosing science-related major at universities, the lack of education system that encourages creative thinking, etc. It is necessary that the accelerated education play an important role to maximize their hidden potentials. It remains to be seen in what places these six participants will find themselves and of what ideas they will come up within the next 17 years. It will be worthwhile to wait until then for another survey.

• Journal of Gifted/Talented Education
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• v.11 no.2
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• pp.127-150
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• 2001

The upcoming century is a knowledge based society which did not exist before which requires creative ability to solve problems. Therefore, it is necessary to Provide a creative program of problem solution in order to match this global trend The creativity of problem solution means the ability to solve a problem using previous ideas in an advanced way or develop new ideas. Creative education is especially important for infants. Because the young mind is where fresh ideas preside and can frame-work the early stage of life like a blank sheet of paper. The Infant-Early Child Creative Development Institute. as an adhesive institute at Yeungjin College, develops various programs that integrate methods which match current trend in this era and also start the Creative Promotion Test with 2,000 Families for the expansion of creative education from the baseline as an alternative method. The infants tested in the creative test will find ways of problem solution through animation beam projects for their given situation and also discuss the problems with their family members. Through these processes the infant and family members will complete the creative structures to solve the problems using limited materials given by the institute, and the final product will be evaluated as objective results. The final evaluation of the test will also be considered the teamwork of family cooperation and the attitudes of participants as well as the product of problem solution. The criterion of the evaluation is to be considered both a creative way of thinking and creative attitudes. Because the score counts were conducted manually it delayed the selection of awarded students who took the test. Also, we found that some parents have difficulty in accessing information to find the score through homepage from the computer. this Problem might be corrected in the future plan. Like Freud's saying, if human character and exploring attitudes during the early stage of a child, a person's creativity is composed their infant period as their basic foundation. Therefore, the family wh first environment the infant encounters will be treated as a prima when making basic structure. From this viewpoint, this creative test work as a festival of creativity fare with 2,000 families.

• Korean Journal of Food Science and Technology
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• v.35 no.5
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• pp.980-987
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• 2003

In this report, we investigated the detoxication effects of Saururus chinenis, Geranium nepalense, Lonicera japonica, Cassia obtusifolia, Glycyrrhiza uralensis, or their mixtures by employing acute toxicity tests for nicotine and dioxin. When fatal doses $(LD_{100}\;=\;42\;mg/kg)$ of nicotine were injected into the abdominal cavities of ICR mice, those treated with OHEM showed delayed paralysis, half the duration of hyperactivity, and a 73 % survival rate. The results revealed the strong detoxicating effects of the mixtures. We also measured the amount of the degradation product of nicotine and cotinine in humans. Consumption of OHEM promoted (he more specific) the metabolic pathways of nicotine, increasing continine excretion by 1.5 times. As a result the amount of cotinine in urine was reduced to less than 5% after treatment with OHEM. In order to test the toxicity of dioxin, we used TcnN(SD)BR rats exposed to TCDD. While TCDD treatment reduced the blood levels of hemoglobin and platelet, OHEM consumption relieved these effects and, furthermore, helped to recover the number of platelet to the normal level (p<0.05). Moreover, neutrophils (%) and monocytes (%), which were reduced by the injection of TCDD, recovered to normal levels upon treatment with OHEM. The amount albumin reduced by TCDD (p<0.05) normalized, while the activities of GOT and GTP increased by TCDD were reduced. Increases in total cholesterol and neutral fatty acids induced by TCDD were also reduced by OHEM injection (p<0.05). In the kidney, TCDD-induced rises in creatinine were suppressed by OHEM treatment, while decreases in iron levels from TCDD were raised to normal. The treatment of TCDD had more toxic effects in the blood and pancreas than on the liver, kidney and heart. On the other hand, the detoxication of OHEM had significant effects on the liver and pancreas. The normalization by OHEM of various clinical abnormalities induced by TCDD demonstrates the detoxicating effect of OHEM that ameliorates systemic metabolism not properly functioning.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.5
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• pp.455-461
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• 2002

limiting nutrients for phytoplankton were determined by dissored inorganic nitrogen/phosphorous (DIN/DIP) in situ and algal bioassay experiment in the Seomjin River estuary during a study period from March 1999 to October 2001. DIN/DIP ranged from 14.7 to 681.1 during the study period. DIN/DIP was over 16 at the upper and middle estuarine region where salinity was lower than 25 psu and chlorophyll a concentration was high, probably indicating P-limitation in this region while the ratio was less than 16 at the high saline (> 25 psu) region, reflecting the supply of DIP from Gwangyang Bay and thereby indicating N-limitation at the lower estuarine region. These results suggested that the spatial distribution of DIN/DIP in the study region was controlled by the high supply of phosphate from Gwangyang Bay, the low input from Seomjin River and the active uptake by phytoplankton within the estuarine system. The bioassay experiments using Skeletonema costatum, Thalaasiosira rotula and in situ phytoplankton assemblage displayed relatively higher growth of phytoplankton in the P-added culture media, indicating P-limitation. This result was well consistent with the spatial distribution of inorganic nutrients. S. costatum showed a rapid adaptation to the low salinity compared to other phytoplankton species. This phenomenon seemed to account for the strong (> $90\%$ in total cell number) S. costatum bloom in autumn in this estuary. Moreover, although phytoplankton growth rate was higher in the P-added culture media at the end of culture experiment of in situ phytoplankton, the fast growth in the trace metal-added media at the beginning of the experiment suggested a possibility of limitation by other micro-nutrients such as trace metal and vitamin etc.

• Journal of the Korean Geographical Society
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• v.29 no.4
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• pp.402-419
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• 1994

Telecommunication and intormation technology have been conceived as crucial as well as revolutionary elements for recent and future social and economic development, and their development have led to a spatial reorganization and locational change of economic activities. Information technology has resulted in important changes in the organization structure and location of firm. This study draws attention to the understanding of the relationship between the diffusion of information technology and changes in firm activities with the special reference to the service industry of the United States. Information technology has had a significant impact on the growth and changes of the service industry of the United States through changes in the organizational and employment structure, market structure, and locational changes. The impact of information technology on location changes of the service industry shows two opposite patterns, concentration and decentralization. Among these patterns, the location change in the service industry of the United States reveals predominantly the decentralization tendency such as suburbanization and transfer to lower ranking cities rather than concentration. In case of Korea, however, it is anticipated that the rapid development of information technology may lead to the concentration of the service industry in Seoul and Capital region.