• Proceedings of the Korean Society for Applied Microbiology Conference
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• 한국미생물생명공학회 2001년도 Proceedings of 2001 International Symposium
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• pp.62-72
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• 2001

The N-carbamoyl-D-amino acid amidohydrolase (D-carbamoylase) gene (dcb) from Agrobacterium tumefaciens AM 10 has been successfully cloned and expressed in Escherichia coli. Expression of D-carbamoylase gene under the 17 promoter in different host strains showed that the optimal expression was achieved in E. coli JM109 (DE3) with a 9-fold increase in enzyme production compared to the wild-type strain. The co-expression of the GroEL/ES protein with D-carbamoylase protein caused an in vivo solubilization of D-carbamoylase in an active form. The synergistic effect of GroEL/ES at 28$^{\circ}C$ led to 60 ％ solubilization of the total expressed target protein with a 6.2-fold increase in enzyme activity in comparison to that expressed without GroEL/ES and 43-fold increase in enzyme activity compared to A. tumefaciens AM 10. Attempts to express D-carbamoylase in an altered redox cytoplasmic milieu did not improve the enzyme production in an active form. The Histidyl-tagged D-carbamoylase was purified in a single step by Nickel-affinity chromatography and was found to have a specific activity of 9.5 U/mg protein.

• Journal of the Korean Applied Science and Technology
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• 제8권1호
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• pp.69-78
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• 1991

Frog legs were stored before and after 30, 60, 90, 120 days at $-30^{\circ}C$ and $-70^{\circ}C$. The amounts of triglycerides, glycolipide and phospholipids by TLC and composition of fatty acids by GC were shown in this experiment, there fore, I have got these following results. 1. The amount of PL in frog legs consisted of 80% of the total lipids before storage. As the days of storage was longer, the amount of TG was increased, while PL and GL were decreased. 2. TG and ES & HC were composed of 66% to 77% of the neutral lipids composition before storage. As the days of storage was longer, the amount of MG, DG. FFA and ES & HC were increased, while FS and TG were decreased. 3. PE and PC were composed of 78% to 81% of the phospholipids composition before storage. As the days of storage was longer, the amount of FA and PS were increased, while PC and PE were decreased. 4. The composition of fatty acids in neutral lipids of frozen frog legs at $-30^{\circ}C$, 120 days was studied, Capric acid. Stearic acid, Behnic acid. Oleic acid and Linoleic acid were increased as compared with initial neutral lipids.

• Journal of the Korean Society of Food Science and Nutrition
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• 제27권2호
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• pp.207-213
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• 1998

These experiments were carried out to investigate the effects of electrical stimulation(ES) and delayed chilling (DC) on the quality characteristics of Hanwoo beef. The left half carcass was treated with ES(550V, 90sec)within postmortem 30min. The electrical stimulated half carcass was subjected to chilling at 16$^{\circ}C$ for 24hr, and then stored at 2$\pm$2$^{\circ}C$ for 15days (ESDC). The right half carcass was stored at 2$\pm$2$^{\circ}C$ for 15 days (NES). ESDC showed a rapid pH fall and tended to reach to pH5.54 at postmortem 2 hrs. But, there was no consistent effect of electrical stimulation and delayed chilling on meat color, cooking loss and water holding capacity. Myofibril fragmentation index was higher than that of NES during storage. ESDC showed lower shear force value and strength consistently than NES. SDS-PAGE band patterns of myofibrils showed the rapid breakdown of troponin T and troponin I band in ESDC, compared with NES, and revealed the specific band below myosin light chain-2 pattern in ESDC.

• Journal of Korean Academy of Dental Administration
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• 제11권1호
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• pp.38-46
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• 2023

Composite resins, commonly used in clinical practice, have been developed to improve aesthetics to obtain smooth surfaces. Although the restored composite resin has a smooth surface, it gradually becomes rough over time. Therefore, this study measured glossiness to evaluate the surface of various composite resins and attempted to evaluate the maintenance of glossiness of composite resins by observing surfaces that change to roughness. Specimens were produced using resin used in clinical practice: Gradia direct anterior (GA), Tetric N-Ceram (TN), Ceram.X Sphere TEC one (CX), Filtek Z350XT (FT), Estelite sigma quick (ES). After creating a smooth surface with slide glass, five locations were randomly selected to measure surface gloss, and the average was the representative value of the specimen. Roughness was applied to the specimen under water pouring at the same speed and pressure using SiC paper #2400, 1200, and 400. The gloss unit of different SiC papers was measured. To evaluate the gloss unit and gloss retention between composite resins, one-way analysis of variance and Tukey multiple comparisons test were used. As a result of the study, there was a difference in gloss unit of specimens produced under the same conditions. Although the degree differed depending on the composite resin, there was also a difference in gloss retention. Based on the findings, composite resins show differences in gloss due to their different characteristics. Ceram.X Sphere TEC one (CX) showing the lowest gloss retention and Estelite sigma quick (ES) showing the highest.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.104-104
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• 2003

As an effort to direct differentiation of human embryonic stem (hES, MB03) cells to dopamine-producing neuronal cells, Nurr1 was transfected using conventional transfection protocol into MB03 and examined the expression of tyrosine hydroylase (TH) after differentiation induced by retinoic acid (RA) and ascorbic acid (AA). Experimentally, cells were transfected with linearized Nurr1 cDNA in pcDNA3.1 (+)-hygovernight followed by selection in medium containing hygromycin-B (150 $\mu$/ml). Expression of Nurr1 mRNA was confirmed by RT-PCR and protein by immunocytochemistry in the drug resistant clones. In order to study the effect of Nurr1 protein on the differentiation pattern of ES cells, one of the positive clones (MBNr24) was allowed to form embryoid body (EB) for 2 days and were induced to differentiate for another 4 days using RA (1 $\mu M$) and AA (50 mM) (2-/4+ protocol) followed by selection in N2 medium for 10 or 20 days. After 10 days in N2 medium, cells immunoreactive to anti-GFAP, anti-TH, or anti-NF200 antibodies were 38.8%, 11%, and 20.5%, respectively. After 20 days in N2 medium, cells expressing GFAP, TH, or NF200 were 28%, 15% and 44.8%, respectively but approximately 9% of MB03 expressed TH protein when the cells were induced to differentiate using a similar prorocol, These results suggest that ectopic expression of Nurr1 enhances generation of TH+ cells as well as neuronal cells when hES cells were differentiated by 2-/4+ protocol.

• Proceedings of the Korean Geotechical Society Conference
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• 한국지반공학회 2010년도 춘계 학술발표회
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• pp.1148-1155
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• 2010

To suggest a modified shallow foundation design method which can be considered the scale effect of foundation on IGM(intermediate geomaterial) soil layer, the weathered soil layer that is uniformly formed up to 8m(2B) with over 50 N-value is selected and 3 times field loading tests are performed on several sized square-shaped shallow foundations with 30, 75, 150, 240 and 400cm in width respectively. Because the soil modulus of elasticity(Es) calculated by soil investigation and 1st field test(PBT) results showed an underestimated tendency, a modified correlation is required for the reasonable estimation of Es on the weathered soil. Also, the N-value was increased with an increasing in depth. However, the N-values around the test foundations showed the different values even though the foundations on the same level because the test site was arranged by excavation. Therefore, the more detail soil investigations are required for the each test foundations respectively. Since Es based on elasticity theory is determined by the stress distribution shape of the foundation and elasticity modulus of the soil, the scale effect considered pressure-settlement curve can be clearly derived from the correlation on stress distribution shape and the variation of soil elasticity modulus with depth. Therefore, the modified correlation will be suggested to estimate a reasonable Es on the weathered soil, and the scale effect considered shallow foundation design method is also developed based on the elastic theory and field tests in this research.

• Journal of Community Nutrition
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• 제8권2호
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• pp.102-106
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• 2006

The purposes of this study were to investigate the effects of the popularity of menu items in nutrient consumption of school-aged children participating in a School Breakfast Program. The weighed plate waste method was used to determine the nutrient intake of students. The nutrient intake was evaluated based on the popularity of the menu item, gender, and grades. The average intakes of all nutrients except energy, fiber, and sodium were well within the goals. Actual nutrient intake varied based upon the popularity of $entr{\acute{e}}es$ and the popularity of menu items was a main effect in nutrient content of meals. When the most popular $entr{\acute{e}}es$ were served, school-aged children's energy intake and School Breakfast Program participation rate increased. Saturated fat and sodium intakes also were higher than the goal when the most popular $entr{\acute{e}}es$ were served. The significant main effect was grades for total fat (p < 0.05) and calcium (p < 0.05), which was qualified by the two-way interaction between gender and grades for saturated fat (p < 0.05), protein (p<0.1), iron (p<0.01), vitamin C (p<0.01) and carbohydrates (p<0.001). Gender itself was not a significant main effect. Based on the findings, the suggestions for educating school-aged children on more healthful breakfast food choices and reformulating recipes for the popular $entr{\acute{e}}es$ are made. (J Community Nutrition 8(2): 102-106, 2006)

• Proceedings of the Korean Society of Developmental Biology Conference
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.101-101
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• 2003

Main strategy for a treatment of Parkinson's disease (PD), due to a progressive degeneration of dopaminergic neurons, is a pharmaceutical supplement of dopamine derivatives or ceil replacement therapy. Both of these protocols have pros and cons; former exhibiting a dramatic relief but causing a severe side effects on long-term prescription and latter also having a proven effectiveness but having availability and ethical problems Embryonic stem (ES) cells have several characteristics suitable for this purpose. To investigate a possibility of using ES cells as a carrier of therapeutic gene(s), human ES (hES, MB03) cells were transfected with cDNAs coding for tyrosine hydroxylase (TH) in pcDNA3.1 (+) and the transfectants were selected using neomycin (250 $\mu /ml$). Expression of TH being confirmed, two of the positive clone (MBTH2 & 8) were second transfected with GTP cyclohydrolase 1 (GTPCH 1) in pcDNA3.1 (+)-hyg followed by selection with hygromycin-B (150 $\mu /ml$) and RT-PCR confirmation. By immune-cytochemistry, these genetically modified but undifferentiated dual drug-resistant cells were found to express few of the neuronal markers, such as NF200, $\beta$-tubulin, and MAP2 as well as astroglial marker GFAP. This results suggest that over-production of BH4 by ectopically expressed GTPCH I may be involved in the induction of those markers. Transplantation of the cells into striatum of 6-OHDA- denervated PD animal model relieved symptomatic rotational behaviors of the animals. Immunohistochemical analyses showed the presence of human cells within the striatum of the recipients. These results suggest a possibility of using hES cells as a carrier of therapeutic gene(s).

• KSBB Journal
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• 제23권5호
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• pp.403-407
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• 2008

Recently, Candida antarctica lipase B (CalB) draws attention from industries for various applications for food, detergent, fine chemical, and biodiesel, because of its characteristics as an efficient biocatalyst. Since many industrial processes carry out in organic solvent and at high temperature, CalB, which is stable under harsh condition, is in demand from many industries. In order to reform CalB promptly, the expression system which has advantages of ease to use and low cost for gene libraries screening was developed using E. coli. The E. coli strains, Rosettagami with competence for enhanced disulfide bond formation, Novablue, and $DH5{\alpha}$, were exploited in this study. To obtain the soluble CalB, the pCold I vector expressing the cloned gene at $15^{\circ}C$ and the chaperone plasmids containing groES/groEL, groES/groEL/tig, tig, dnaK/dnaJ/grpE, and dnaK/dnaJ/grpE/groES/groEL were used for coexpression of CalB and chaperones. The colonies expressing functional lipase were selected by employing the halo plate containing 1% tributyrin, and the CalB expression was confirmed by SDS-PAGE. E. coli Rosettagami and $DH5{\alpha}$ harbouring groES/groEL chaperones were able to express soluble CalB effectively. From a facilitative point of view, E. coli $DH5{\alpha}$ is more suitable for further mutation study.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.95-95
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• 2002

Human embryonic stem (hES) cells can be induced to differentiate into tyrosine hydroxylase expressing (TH+) cells that may serve as an alternative for cell replacement therapy for Parkinson's disease (PD). To examine in vitro differentiation of hES (MB03, registered in NIH) cells into TH+ cells, hES cells were induced to differentiate according to the 4-/4+ protocol using retinoic acid (RA), ascorbic acid (AA), and/or lithium chloride (LiCl) followed by culture in N2 medium for 14 days, during which time the differentiation occurs. Immunocytochemical stainings of the cells revealed that approximately 21.1% of cells treated with RA plus AA expressed TH protein that is higher than the ratio of TH+ cells seen in any other treatment groups (RA, RA+LiCl or RA+AA+LiCl). In order to see the differentiation pattern in vivo and the ability of in vitro differentiation-induced cells in easing symptomatic motor function of PD animal model, cells (2 $\times$ 10$^{5}$ cells/2${mu}ell$) undergone 4-/4+ protocol using RA plus AA without any further treatment were transplanted into unilateral striatum of MPTP-lesioned PD animal model (C57BL/6). Following the surgery, motor behavior of the animals was examined by measuring the retention time on an accelerating rotar-rod far next 10 weeks. No significant differences in retention time of the animals were noticed until 2 weeks post-graft; however, it increased markedly at 6 weeks and 10 weeks time point after the surgery. Immunohistochemical studies confirmed that a reasonable number of TH+ cells were found at the graft site as well as other remote sites, showing the migrating nature of embryonic stem cells. These results suggest that in viかo differentiated hES cells relieve symptomatic motor behavior of PD animal model and should be considered as a promising alternative for the treatment of PD.