Proceedings of the Korean Society of Developmental Biology Conference
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2003.10a
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pp.123-123
/
2003
This study was conducted to determine the ability of nuclear development of canine oocytes depend on the kind of maturation media and addition of serum sources. Ovaries were collected from a bitches at various stages of estrus cycle by an ovariohysterectomy. Oocytes were collected of cumulus oocytes complexes after slicing of ovaries with blade. The maturation medium was containing 0.6 mM/ml cysteine, 0.2 mM pyruvic acid, 20 ng/ml $E_2$ and 1 $\mu g/ml$ rbST Exp. 1, the oocytes were matured in four different maturation medium as follows: 1) TCM-199, 2) DMEM, 3) NCSU37 and 4) modified-NCSU37 with 10% FBS. Exp. 2: the oocytes were matured in mNCSU37 supplemented with different protein sources (10% FBS, 10% EDS, 0.3% BSA and 0.1% PVA) to select the optimal one. Oocytes were matured in a humidified atmosphere containing 5% $CO_2$ at $39{\circ}C$ for 72 hrs. The maturation rate were analyzed by Duncan's multiple range test using General Linear Models procedure in SAS. The rates of meiotic resumption to MI-MII depend on different culture media were achieved with TCM-199 (5.2%), DMEM (5.0%), NCSU37 (7.2%) and m-NCSU37 (5.9%), respectively. The rates of meiotic resumption to MI-MII according to addition of protein source were 10% FBS (13.3%), 10% EDS (25.0%), 0.3% BSA (25.0%) and 0.1% PVA (15.4%), respectively. In conclusion, the results obtained showed that in vitro maturation media and protein supplement to m-NCSU37 culture medium tested did not promote the final steps of IVM in canine oocytes.
Background: Inactivation of tumor suppressor genes is believed to be important in the development of many human malignancies. Recently, several lines of evidence have indicated that the wild type p53 gene located at 17p13.3, may function as a tumor suppressor gene and that a mutant p53 gene could promote transformation by inactivating normal p53 function in a dominant negative fashion. These broad spectrum of p53 mutation in human cancers provide that mutant p53 and their protein may be potential targets of tumor diagnostic and therapeutic interventions. Method: Colony formation was performed to investigate growth suppressional ability. p53 expression pattern was examined by western blot and p53-mediated transactivation ability was assessed by CAT activity. SNU C2A cells were observed in apoptotic aspects induced by etoposide and $H_2O_2$ treatment, detecting sensitivity on agent, DNA fragmentation through agarose gel, chromatin condensation by fluorescence microscope, and cell cycle distribution by FACS. Result: 1) p53 mutant his179arg ($histidine{\rightarrow}arginine$) detected in SNU C2A cells lost transcriptional activity and growth suppression ability, showing dominant negative effect on its wild type p53. 2) Etoposide-treated SNU C2A cells induced apoptosis, exhibiting dramatic reduction of cell growth, DNA fragmentation, nuclear condensation formation of apoptotic body and increment of sub-G1 cell fraction. 3) Etoposide and $H_2O_2$-treated SNU C2A cells have no high increase of p53 expression and overexpressed p53 protein changed localization, from cytoplasm to nucleus. Also, p53-mediated transcriptional activity was increased by agents-treatment. Conclusion: SNU C2A cells coexpress wild-type and mutant p53 protein induced apoptosis in the condition on DNA damage, through localizational shift from cytoplasm to nucleus of p53 protein rather than the induction of p53 protein. SNU C2A cells derived mutant p53 his179arg abrogated both the growth supression ability and transactivational activity, showing inhibition effect on transcriptional activity of wild type p53, but did not repress the activity of wild type p53 in SNU C2A cells owing to dominant activity of wild type. These cell condition may provide new gene therapeutic implications leading effective antiproliferation of cell when mutant and wild-type p53 protein were co-expressed in cell.
The core price policy of on-line game marketing are FPP(Fixed Pre Paid model and PPU(Pay Per Use) model. These two models have been a on-line game company's billing system and a fundamental of MMORPG in Korea. However, they took root billing system only for first movers recently. In now, the market share of several first movers is exceeding 80%, late movers witch have same billing system cannot take part in pair competition. Even though in MMORPG, many games of late movers were favorably noticed by a lot of gamers during Evaluation. Test, a lot of companies are bankrupt before make business. Late Movers declare free game first thing, they maintain their existence and win over customers in on-line game market. And next, they guarantee item selling, give multiple experience value and game money, at last, induce their customers to pay service. As it makes trouble between pay user and free user, and it linked up with the collapse of game contents balance that designed for FPP billing system, And then meet unexpected result which reduction of game life cycle. In this Paper, we classified several contents services based on game contents, and suggested contents premium services which adopted low cost strategy lead to micro payment. we hope it will apply to late movers' new billing system in MMORPG.
Journal of the Korean Society of Food Science and Nutrition
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v.38
no.7
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pp.964-969
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2009
To improve hygienic quality of vegetable fresh juice containing non-thermal process materials, an irradiation technology ($0{\sim}5\;kGy$) was applied and microbiological and physicochemical changes were evaluated during storage. The initial number of total aerobic bacteria of vegetable fresh juice containing non-thermal process materials was $4.8{\times}10^3\;CFU/mL$ and the number was increased during storage. At day 7, the number reached $3.0{\times}10^5\;CFU/mL$. However, the samples gamma-irradiated at $1{\sim}5\;kGy$ were $1.2{\times}10^2{\sim}1.0{\times}10^3\;CFU/mL$, which was lower than that of control and higher than 3 kGy irradiation, showing more than a decimal reduction. The lightness and yellowness of irradiated sample was higher than control but no difference was found among storage days; whereas, redness was decreased by gamma irradiation with no difference found among storage days. Ascorbic acid and carotenoid contents were decreased by irradiation treatment and increase of storage days. There were no changes in flavonoid contents; however, the content of polyphenols was increased by irradiation of sample.
Transactions of the Korean Society of Automotive Engineers
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v.24
no.2
/
pp.144-151
/
2016
This study effect of engine oils on regulated fuel economy and emissions including particulate matter (PM) to provide basic data for management of engine oil in vehicles. Three engine oils (Group III base oil, Group III genuine oil with additive package and synthetic oil with poly alpha olefins (PAOs)) were used in one gasoline, one LPG(liquefied petroleum gas) and two diesel vehicles. In the case of diesel vehicles, one is a diesel vehicle without DPF (diesel particulate filter) other is a diesel vehicle with DPF. In this study, the US EPA emission test cycle FTP-75, representing city driving, was used. HORIBA, PIERBURG, and AVL gas analyzers were used to measure the fuel economy and regulated emissions such as CO, NOx, and THC. The number of PM was measured using a PPS (pegasor particle sensor). And, the shape of PMs was analyzed by SEM (scanning electron microscope). The effects of oil type on fuel economy, exhaust gas, and PM were not significant because engine oil consumption by evaporation and combustion in the cylinder is very tiny. Fuel and vehicle type were dominant factors in fuel economy and emissions. HC emission from gasoline vehicles was higher than that from other vehicles and NOx emission from diesel vehicles was higher than that from other vehicles. The number of PM was not affected by the engine oil, but by the driving pattern and fuel. The shapes of the PM, sampled from each vehicle using any test engine oil, were similar.
Effect of culture conditions on the fermentation of wheat flour solution by mixed lactic acid bacteria of Lactobacillus brevis, L. fermentum and L. plantarum was investigated. The optimum temperature for the fermentation of wheat flour solution was $35^{\circ}C$ because pH decreased the lowest value and TTA (total titrable acidity) increased the highest value at this temperature. In aerobic condition, fermentor was purged with air at 1.0 vvm and was purged with nitrogen gas at 1.0 vvm in anaerobic condition. The decrease of pH and the increase of TTA in aerobic condition were higher than those in anaerobic condition. In aerobic condition, the optimum condition of oxygen supply was found to be oxygen transfer rate coefficient of $60\;hr^{-1}$ which corresponded to agitation speed of 250 rpm in a 5 L fermentor. Repeated fed-batch cultures were performed using pH-stat in order to increase the productivity of fermented wheat flour. With increasing the repeated fraction of culture volume, mean cycle time increased but maximum operation time decreased. However, the volume of produced broth per culture volume per time and total volume of produced broth per culture volume were maximum at the repeated fraction of culture volume of 20%. In a repeated fed-batch fermentation of wheat flour solution using mixed lactic acid bacteria, the culture condition was optimum at temerature of $35^{\circ}C$, aeration rate of 1.0 vvm, oxygen transfer rate coefficient of $60\;hr^{-1}$, and repeated fraction of culture volume of 20%.
Lee Han Chang;Yeam Mi Jung;Kim Gun Ho;Choi Kang Duk;Lee Seoung Hee;Shim Insop;Lee Hye Jung;Hahm Dae Hyun
Journal of Physiology & Pathology in Korean Medicine
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v.17
no.6
/
pp.1393-1403
/
2003
The genetic effects of restraint stress challenge on HPA axis and the therapeutic effect of Boshimgeonbi-Tang on the stress were studied with cDNA microarray analyses on hypothalamus using an immobilization-stress mouse as stress model. Male CD-1 mice were restrained in a tightly fitted and ventilated vinyl holder for 2hours once a day, and this challenge was repeated for seven consecutive days. The body weights of the immobilization-stress mice were diminished about 25 percent degree as compared to normal ones. Seven days later, total RNA was extracted from the organs of the mouse, body-labeled with CyDye/sup TM/ fluorescence dyes (Amersham Bioscience Co., NJ), and then hybridized to cDNA microarray chip. Scanning and analyzing the array slides were carried out using GenePix 4000 series scanner and GenePix Pro/sup TM/ analyzing program, respectively. The expression profiles of 109 genes out of 6000 genes on the chip were significantly modulated in hypothalamus by the immobilization stress. Energy metabolism-, lipid metabolism-, apoptosis- and signal transduction-related genes were transcriptionally activated whereas DNA repair-, protein biosynthesis-, and structure integrity-related genes were down-regulated in hypothalamus. The 58 genes were up-regulated by the mRNA expression folds of 1.5 to 7.9. and the 51 genes were down-regulated by 1.5 - 3.5 fold. The 20 genes among them were selected to confirm the expression profiles by RT-PCR. The mRNA expression levels of Tnfrsf1a (apoptosis), Calm2 (cell cycle), Bag3 (apoptosis), Hspe1 (protein folding), Aatk (apoptosis), Dffa (apoptosis), Itgb1 (cell adhesion), Vcam1 (cell adhesion), Fkbp5 (protein folding), BDNF (neuron survival) were restored to the normal one by the treatment of Boshimgeonbi-Tang.
These study was carried out to investigate the effects of the recovery time, diameter of oocytes on in vitro fertilization or intracytoplasmic sperm injection (ICSI). The in vitro maturation rates to MII stage of oocytes recovered at the inactive, follicular and luteal stages matured for 72 h were $1.4{\pm}0.0%$, $43.4{\pm}3.2%$ and $10.8{\pm}2.7%$, respectively. The fertilization rates of in vitro cultured oocytes recovered from ovaries at the in active, follicular and luteal stages were $0.0{\pm}0.0%$, $15.7{\pm}3.4%$ and $7.6{\pm}3.5%$, respectively. The in vitro maturation rate of oocytes recovered from ovaries at the follicular stage of the reproductive cycle was significantly higher than those at the inactive and luteal stages (p<0.05). The penetration rate determined that the percentages of oocytes with diameters in the < $100\;{\mu}m$, 100 to $100\;{\mu}m$ and 110 to $120\;{\mu}m$ ranges were $17.5{\pm}4.7%$, $43.9{\pm}4.5%$, $21.3{\pm}3.4%$, respectively. The penetration rate of oocytes with diameters between 100 to $100\;{\mu}m$ was significantly higher than that of oocytes whose diameters were < $100\;{\mu}m$ and $110{\sim}120\;{\mu}m$ (p<0.05). The penetration rate of oocytes determined that the percentages of ovaries with diameters between 1 to 5 mm and 6 to 10 mm were $32.9{\pm}3.2%$ and $17.5{\pm}3.7%$, respectively. Thus, the diameters of the ovaries were significantly higher at 1 to 5 mm (p<0.05). A total of 264 oocytes were fixed and stained after co-incubation with sperm, of which 72 had identifiable nuclear material. After in vitro fertilization for 20 hrs, 27.3% of oocytes were penetrated by spermatozoas. Oocytes were fixed and stained after ICSI, of which 38 oocytes contained identifiable nuclear material. After in vitro fertilization and ICSI for 20 hrs, to 27.3% and 67.9% of oocytes were penetrated by spermatozoas. The in vitro fertilization rates by ICSI was significantly higher than that in vitro fertilization method (p<0.05).
Park, Soon-Jung;Jeon, Young-Joo;Kim, Ju-Mi;Shin, Jeong-Min;Chae, Jung-Il;Chung, Hyung-Min
Reproductive and Developmental Biology
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v.34
no.3
/
pp.143-151
/
2010
Pluripotency of human embryonic stem cell (hESC) is one of the most valuable ability of hESCs for applying cell therapy field, but also showing side effect, for example teratoma formation. When transplant multipotent stem cell, such as mesnchymal stem cell (MSC) which retains similar differentiation ability, they do not form teratoma in vivo, but there exist limitation of cellular source supply. Accordingly, differentiation of hESC into MSC will be promising cellular source with strong points of both hESC and MSC line. In this study, we described the derivation of MSC like cell population from feeder free cultured hESC (hESC-MSC) using direct differentiation system. Cells population, hESC-MSC and bone marrow derived MSC (BM-MSC) retained similar characteristics in vitro, such as morphology, MSC specific marker expression and differentiation capacity. At the point of differentiation of both cell populations, differentiation rate was slower in hESC-MSC than BM-MSC. As these reason, to verify differentially expressed molecular condition of both cell population which bring out different differentiation rate, we compare the molecular condition of hESC-MSC and BM-MSC using 2-D proteomic analysis tool. In the proteomic analysis, we identified 49 differentially expressed proteins in hESC-MSC and BM-MSC, and they involved in different biological process such as positive regulation of molecular function, biological process, cellular metabolic process, nitrogen compound metabolic process, macromolecule metabolic process, metabolic process, molecular function, and positive regulation of molecular function and regulation of ubiquitin protein ligase activity during mitotic cell cycle, cellular response to stress, and RNA localization. As the related function of differentially expressed proteins, we sought to these proteins were key regulators which contribute to their differentiation rate, developmental process and cell proliferation. Our results suggest that the expressions of these proteins between the hESC-MSC and BM-MSC, could give to us further evidence for hESC differentiation into the mesenchymal stem cell is associated with a differentiation factor. As the initial step to understand fundamental difference of hESC-MSC and BM-MSC, we sought to investigate different protein expression profile. And the grafting of hESC differentiation into MSC and their comparative proteomic analysis will be positively contribute to cell therapy without cellular source limitation, also with exact background of their molecular condition.
Kim, Yo-Sup;Hong, Yeong-Pyo;Cho, Young-Il;Kim, Jin-Su;Eun, Jong-Won;Lee, Jong-Yong;Lee, Sang-Hun
Journal of the Korea Academia-Industrial cooperation Society
/
v.11
no.9
/
pp.3472-3480
/
2010
The Clustering technology of Energy efficiency wireless sensor network gets the energy efficiency by reducing the number of communication between sensor nodes and sink node. In this paper, First analyzed on the clustering technique of the distributed clustering protocol routing scheme LEACH (Low Energy Adaptive Clustering Hierarchy) and HEED (Hybrid, Energy-Efficient Distributed Clustering Approach), and based on this, new energy-efficient clustering technique is proposed for the cause the maximum delay of dead nodes and to increase the lifetime of the network. In the proposed method, the cluster head is elect the optimal efficiency node based on the residual energy information of each member node and located information between sink node and cluster node, and elected a node in the cluster head since the data transfer process from the data been sent to the sink node to form a network by sending the energy consumption of individual nodes evenly to increase the network's entire life is the purpose of this study. To verify the performance of the proposed method through simulation and compared with existing clustering techniques. As a result, compared to the existing method of the network life cycle is approximately 5-10% improvement could be confirmed.
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