• Journal of Life Science
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• v.24 no.4
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• pp.343-351
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• 2014

Phosphoinositide 3-kinase (PI3K) plays a central role in cell signaling and leads to cell proliferation, survival, motility, exocytosis, and cytoskeletal rearrangements, as well as specialized cell responses, superoxide production, and cardiac myocyte growth. PI3K is divided into three classes; type I PI3K is preferentially expressed in leukocytes and activated by ${\beta}{\gamma}$ subunits of heterotrimeric G-proteins. In this study, the cDNAs encoding the $PI3K{\gamma}$ gene were isolated from a brain cDNA library constructed using the flounder (Paralichthys olivaceus). The sequence of the isolated $PI3K{\gamma}$ was 1341 bp, encoding 447 amino acids. The nucleotide sequence of the $PI3K{\gamma}$ gene was analyzed with that of other species, including Oreochromis niloticus and Danio rerio, and it turned out to be well conserved during evolution. The $PI3K{\gamma}$ gene was subcloned into the expression vector pET-44a(+), and expressed in the E. coli BL21 (DE3) codon plus cell. The resulting protein was expressed as a fusion protein of approximately 49 kDa containing a C-terminal six-histidine extension that was derived from the expression vector. The expressed protein was purified to homogeneity by His-tag affinity chromatography and showed enzymatic activity corresponding to $PI3K{\gamma}$. The binding of wortmannin to $PI3K{\gamma}$, as detected by anti-wortmannin antisera, closely followed the inhibition of the kinase activities. The results obtained from this study will provide a wider base of knowledge on the primary structure and characterization of the $PI3K{\gamma}$ at the molecular level.

• Food Science and Preservation
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• v.21 no.1
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• pp.121-128
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• 2014

This study was conducted in order to investigate the change of isoflavone composition (glycoside and bio-active aglycone), and to evaluate the quality characteristics of Cheonggukjang, which was prepared by different bacillus strains. After the 48-hour fermentation, the contents of daidzein, genistein, and glycitein in the Bacillus subtilis HJ18-3 have significantly increased up to approximately $89.06{\pm}3.59$, $10.36{\pm}0.28$, and $101.37{\pm}3.67ug/g$, respectively. The contents of daidzein, genistein, and glycitein in the Bacillus subtilis KACC 15935 were $38.88{\pm}5.39$, $12.58{\pm}2.14$, and $80.13{\pm}0.71ug/g$, respectively. The original content of daidzein was 3.96 ug/g, while genistein and glycitein were not measured. However, the contents of daidzen and genistein in HJ18-3 and in KACC 15935 were decreased. The ${\alpha}$-Amylase and cellulase activities of Chungkookjang in HJ18-3 were higher than in the KACC 15935. The contents of Chungkookjang in HJ18-3 were $29.70{\pm}11.66$ and $4861.3{\pm}388.07unit/g$, respectively. The amino type nitrogen contents and ammonia type nitrogen contents of Chungkookjang in KACC 15935 were higher than in the HJ18-3. These results suggested that it could be used to increase the bioactivity via fermentation with the Bacillus subtilis possessing a ${\beta}$-glucosidase activity with a view towards the development of functional foods.

• Korean Journal of Biological Psychiatry
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• v.14 no.2
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• pp.115-121
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• 2007

Objetives : Identification of target genes for ethanol in neurons is important for understanding its molecular and cellular mechanism of action and the neuropathological changes seen in alcoholics. The purpose of this study is to identify of altered gene expression after acute treatmet of ethanol in rat gliom cells. Methods : We used high density cDNA microarray chip to measure the expression patterns of multiple genes in cultured rat glioma cells. DNA microarrays allow for the simultaneous measurement of the expression of several hundreds of genes. Results : After comparing hybridized signals between control and ethanol treated groups, we found that treatment with ethanol increased the expression of 15 genes and decreased the expression of 12 genes. Upregulated genes included Orthodenticle(Drosophila) homolog 1, procollagen type II, adenosine A2a receptor, GATA bindning protein 2. Downregulated genes included diacylglycerol kinase beta, PRKC, Protein phosphatase 1, clathrin-associated protein 17, nucleoporin p58, proteasome. Conclusion : The gene changes noted were those related to the regulation of transcription, signal transduction, second messenger systems. modulation of ischemic brain injury, and neurodengeneration. Although some of the genes were previously known to be ethanol responsive, we have for the most part identified novel genes involved in the brain response to ethanol.

• Korean Journal of Construction Engineering and Management
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• v.5 no.5 s.21
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• pp.183-192
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• 2004

Even in Korea the number of steel structure buildings that allow internal space and easy change of their layouts in accordance with the purpose of buildings and box-type steel bridges constructed with thick plates with thickness in a rage just from a few $\beta$AE to \$100\beta$AE is increasing these days and therefore, domestic fabrication and processing technology of members for steel structures is being improved at a pace faster than in the past to meet the growing requirements of consumers for high reliability on quality control on the related steel structures. However, most domestic fabricators os steel structures who are turning out their steel products in accordance with the designs prepared by engineering companies in their respective works for the sake of cost cut more than anything else, hesitating to introduce any advanced new technology into themselves. In the case of the steel structure design application for small and mid-size buildings in particular, it is quite meaningful not only for those who are involved in steel structure business, but also for the people working at construction work fields to review the result of the study on the connections of steel structure members deigned to obtain superb quality of steel structures within short period for steel fabrication and erection at fields in economical ways, as there is a glowing tendency seeking standardization of connection of steel structure members as well as whole structure together with the development on design of construction system of buildings including their exterior and interior decoration materials, manufacture of the related members and fabrication technique structure. This paper has been prepared with the aim to review the peculiar characteristics of buildings constructed with the main frames of steel structures and actual cases of the change made ing the connections between steel structure columns and between columns and girder members in order to reduce the work period necessary for fabrication and erection of steel structures at the maximum as well as the some examples of steel structures fabricated through automatic welding by robots for box-type columns in addition to the description of the problems found in the course of fabricating those steel structures, suggesting possible counter-measures to solve them.

• Korean Journal of Dental Materials
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• v.44 no.3
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• pp.207-216
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• 2017

The effect of cooling rate on precipitation hardening of a Pd-Cu-Ga-Zn metal-ceramic alloy during porcelain firing simulation was investigated and the following results were obtained. When the cooling rate was fast (Stage 0), the hardness of the alloy increased at each firing step and the high hardness value was maintained. When the cooling rate was slow (Stage 3), the hardness was the highest at the first stage of the firing, but the final hardness of the alloy after complete firing was lower. The increase in hardness of the specimens cooled at the cooling rate of Stage 0 after each firing step was caused by precipitation hardening. The decrease in hardness of the specimens cooled at the cooling rate of Stage 3 after each firing step was attributed to the coarsening of the spot-like precipitates formed in the matrix and plate-like precipitates. The matrix and the plate-like precipitates were composed of the $Pd_2(Cu,Ga,Zn)$ phase of CsCl-type, and the particle-like structure was composed of the Pd-rich ${\alpha}$-phase of face-centered cubic structure. Through the porcelain firing process, Cu, Ga, and Zn, which were dissolved in Pd-rich ${\alpha}$ particles, precipitated with Pd, resulting in the phase separation of the Pd-rich ${\alpha}$ particles into the Pd-rich ${\alpha}^{\prime}$ particles and ${\beta}^{\prime}$ precipitates composed of $Pd_2(Cu,Ga,Zn)$. These results suggested that the durability of the final prosthesis made of the Pd-Cu-Ga-Zn alloy can be improved when the cooling rate is fast during porcelain firing simulation.

• Clinical and Experimental Reproductive Medicine
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• v.34 no.1
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• pp.19-31
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• 2007

Objective: Pathogenesis of the endometriosis is very complex and the etiology is still unclear. Our hypothesis is that there may be some difference in gene expression patterns between eutopic endometriums with or without endometriosis. In this study, we analyzed the difference of gene expression profile with cDNA microarray. Methods: Endometrial tissues were gathered from patients with endometriosis or other benign gynecologic diseases. cDNA microarray technique was applied to screen the different gene expression profiles from early and late secretory phase endometria of those two groups. Each three mRNA samples isolated from early and late secretory phase of endometrial tissues of control were pooled and used as master controls and labeled with Cy3-dUTP. Then the differences of gene expression pattern were screened by comparing eutopic endometria with endometriosis, which were labeled with Cy5-dUTP. Fluorescent labeled probes were hybridized on a microarray of 4,800 human genes. Results: Twelve genes were consistently over-expressed in the endometrium of endometriosis such as ATP synthase H transporting F1 (ATP5B), eukaryotic translation elongation factor 1, isocitrate dehydrogenase 1 (NADP+), mitochondrial ribosomal protein L3, ATP synthase H+ transporting (ATP5C1) and TNF alpha factor. Eleven genes were consistently down-regulated in the endometriosis samples. Many extracellular matrix protein genes (decorin, lumican, EGF-containing fibulin-like extracellular matrix protein 1, fibulin 5, and matrix Gla protein) and protease/protease inhibitors (serine proteinase inhibitor, matrix metalloproteinase 2, tissue inhibitor of metalloproteinase 1), and insulin like growth factor II associated protein were included. Expression patterns of selected eight genes from the cDNA microarray were confirmed by quantitative RT-PCR or real time RT-PCR. Conclusion: The result of this analysis supports the hypothesis that the endometrium from patients with endometriosis has distinct gene expression profile from control endometrium without endometriosis.

• Journal of Korean Society of Coastal and Ocean Engineers
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• v.30 no.6
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• pp.306-318
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• 2018

In order to evaluate the seismic capacity of massive vertical type breakwaters which have intensively been deployed along the coast of South Korea over the last two decades, we carry out the preliminary numerical simulation against the PoHang, GyeongJu, Hachinohe 1, Hachinohe 2, Ofunato, and artificial seismic waves based on the measured time series of ground acceleration. Numerical result shows that significant sliding can be resulted in once non-negligible portion of seismic energy is shifted toward the longer period during its propagation process toward the ground surface in a form of shear wave. It is well known that during these propagation process, shear waves due to the seismic activity would be amplified, and non-negligible portion of seismic energy be shifted toward the longer period. Among these, the shift of seismic energy toward the longer period is induced by the viscosity and internal friction intrinsic in the soil. On the other hand, the amplification of shear waves can be attributed to the fact that the shear modulus is getting smaller toward the ground surface following the descending effective stress toward the ground surface. And the weakened intensity of soil as the number of attacking shear waves are accumulated can also contribute these phenomenon (Das, 1993). In this rationale, we constitute the numerical model using the model by Hardin and Drnevich (1972) for the weakened shear modulus as shear waves go on, and shear wave equation, in the numerical integration of which $Newmark-{\beta}$ method and Modified Newton-Raphson method are evoked to take nonlinear stress-strain relationship into account. It is shown that the numerical model proposed in this study could duplicate the well known features of seismic shear waves such as that a great deal of probability mass is shifted toward the larger amplitude and longer period when shear waves propagate toward the ground surface.

• Molecules and Cells
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• v.26 no.2
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• pp.175-180
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• 2008

$I{\kappa}B$ kinase (IKK), the pivotal kinase in signal-dependent activation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$), is composed of multiple protein components, including IKK ${\alpha}/{\beta}/{\gamma}$ core subunits. To investigate the regulation of the IKK complex, we immunoaffinity purified the IKK complex, and by MALDI-TOF mass spectrometry identified a splice variant of zinc finger protein 268 (ZNF268) as a novel IKKinteracting protein. Both the full-length and the spliced form of the ZNF268 protein were detected in a variety of mammalian tissues and cell lines. The genes were cloned and expressed by in vitro transcription/translation. Several deletion derivatives, such as KRAB domain (KRAB) on its own, the KRAB/spacer/4-zinc fingers (zF4), and the spacer/4-zinc fingers (zS4), were ectopically expressed in mammalian cells and exhibited had different subcellular locations. The KRAB-containing mutants were restricted to the nucleus, while zS4 was localized in the cytosol. TNF-${\alpha}$-induced NF-${\kappa}B$ activation was examined using these mutants and only zS4 was found to stimulate activation. Collectively, the results indicate that a spliced form of ZNF268 lacking the KRAB domain is located in the cytosol, where it seems to play a role in TNF-${\alpha}$-induced NF-${\kappa}B$ activation by interacting with the IKK complex.

• Horticultural Science & Technology
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• v.33 no.6
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• pp.947-954
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• 2015

To select suitable spray chrysanthemum cultivars for Agrobacterium-mediated transformation, thirty-nine (39) spray cultivars bred in the National Institutes of Korea and a standard cultivar Jinba from Japan were collected and tested for regeneration rate and Agrobacterium infection assays. MS medium with $0.5mg{\cdot}L^{-1}$ IAA and $1.0mg{\cdot}L^{-1}$ BAP was used for shoot regeneration from leaf disks and internodes. The shoot regeneration rate in leaf disks was the highest in cultivar BRM, followed by cultivars VS, WW and YTM. The cultivar JB (Jinba) used as a transformation material in previous reports ranked similarly to cultivars PK and SPP. In shoot regeneration from internodes, the shoot regeneration rate was the highest for cultivar PA, followed by cultivar WW. The infection rate of leaves and internodes of 40 chrysanthemum cultivars with agrobacterium was investigated. Cultivars WPP, YNW, VS, PP, WW, FA, PA and YMN showed the highest infection levels in leaves, whereas cultivars WPP, PA, PK and YNW had the highest infection levels in internodes. Considering all of these results, cultivars VS and WW were the most appropriate for gene transformation of chrysanthemum using leaves, while cultivar PA was for internodes.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.27 no.4
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• pp.314-320
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• 2001

Matrix metalloproteinases have long been viewed as ideal candidates for proteinases that enables tumor cells to permeated basement membrane defenses and invade surrounding tissue. There is growing evidence that the MMPs have an expanded role, as they are important for the creation and maintenance of a microenvironment that facilitates growth and angiogenesis of tumors at primary and metastatic sites. MT-MMPs are not secreted but instead remaining attached to cell surfaces. Although not all of the MT-MMPs are fully characterized, MT-MMPs have important role in localizing and activating secreted MMPs. The MMP genes are transcriptionally responsive to a wide variety of oncogene, growth factors, cytokine, and hormones. Currently, a number of MMP inhibitors are being developed and some have reached clinical trials as anti-metastatic or anti-cancer therapies. MT1-MMP is involved in the activation of proMMP-2. MT1-MMP is significant not only as a tumor marker but as a new target for chemotherapy against cancer. The purpose of this study was to evaluate the effects of protein kinase C inhibitor(genistein) on the proliferation of HT1080 and expression of MT1-MMP mRNA. Human fibrosarcoma cell line HT1080 was cultured and divided 2 groups. The experimental group was treated with $100{\mu}M$ genistein and incubated 12h, 24h for $[3^H]-thymidine$ uptake assay and northern hybridization individually. And the control group was treated with same amount of PBS for the above procedures. $[3^H]-thymidine$ incorporation was measured with ${\beta}$ ray detector. And RT-PCR and northern blotting for MT1-MMP mRNA was performed. The results were as follows 1. $[3^H]-thymidine$ uptake was reduced in experimental group with statistical significance. 2. MT1-MMP mRNA expression was significantly reduced in experimental group. These results showed that protein kinase C inhibitor (genistein) inhibited proliferation of HT1080 and almost completely blocked transcription of MT1-MMP mRNA. So, it is possible to use the protein kinase inhibitor (genistein) as anti-metastatic and anti-proliferative agent.