Kim, Jae Wha;Yoon, Sun Young;Joo, Joung-Hyuck;Kang, Ho Bum;Lee, Younghee;Choe, Yong-Kyung;Choe, In Seong
IMMUNE NETWORK
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v.2
no.3
/
pp.175-181
/
2002
Background: S100A6 is a calcium-binding protein overexpressed in several tumor cell lines including melanoma with high metastatic activity and involved in various cellular processes such as cell division and differentiation. To detect S100A6 protein in patient' samples (ex, blood or tissue), it is essential to produce a monoclonal antibody specific to the protein. Methods: First, cDNA coding for ORF region of human S100A6 gene was amplified and cloned into the expression vector for GST fusion protein. We have produced recombinant S100A6 protein and subsequently, monoclonal antibodies to the protein. The specificity of anti-S100A6 monoclonal antibody was confirmed using recombinant S100A recombinant proteins of other S100A family (GST-S100A1, GST-S100A2 and GST-S100A4) and the cell lysates of several human cell lines. Also, to identify the specific recognition site of the monoclonal antibody, we have performed the immunoblot analysis with serially deleted S100A6 recombinant proteins. Results: GST-S100A6 recombinant protein was induced and purified. And then S100A6 protein excluding GST protein was obtained and monoclonal antibody to the protein was produced. Monoclonal antibody (K02C12-1; patent number, 330311) has no cross-reaction to several other S100 family proteins. It appears that anti-S100A6 monoclonal antibody reacts with the region containing the amino acid sequence from 46 to 61 of S100A6 protein. Conclusion: These data suggest that anti-S100A6 monoclonal antibody produced can be very useful in development of diagnostic system for S100A6 protein.
Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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2009.06a
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pp.327-327
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2009
ZnO with a large band gap (~3.37 eV) and exciton binding energy (~60 meV), is suitable for optoelectronic applications such as ultraviolet (UV) light emitting diodes (LEDs) and detectors. However, the ZnO-based p-n homojunction is not readily available because it is difficult to fabricate reproducible p-type ZnO with high hall concentration and mobility. In order to solve this problem, there have been numerous attempts to develop p-n heterojunction LEDs with ZnO as the n-type layer. The n-ZnO/p-GaN heterostructure is a good candidate for ZnO-based heterojunction LEDs because of their similar physical properties and the reproducible availability of p-type GaN. Especially, the reduced lattice mismatch (~1.8 %) and similar crystal structure result in the advantage of acquiring high performance LED devices. In particular, a number of ZnO films show UV band-edge emission with visible deep-level emission, which is originated from point defects such as oxygen vacancy, oxygen interstitial, zinc interstitial[1]. Thus, defect-related peak positions can be controlled by variation of growth or annealing conditions. In this work, the undoped ZnO film was grown on the p-GaN:Mg film using RF magnetron sputtering method. The undoped ZnO/p-GaN:Mg heterojunctions were annealed in a horizontal tube furnace. The annealing process was performed at $800^{\circ}C$ during 30 to 90 min in air ambient to observe the variation of the defect states in the ZnO film. Photoluminescence measurements were performed in order to confirm the deep-level position of the ZnO film. As a result, the deep-level emission showed orange-red color in the as-deposited film, while the defect-related peak positions of annealed films were shifted to greenish side as increasing annealing time. Furthermore, the electrical resistivity of the ZnO film was decreased after annealing process. The I-V characteristic of the LEDs showed nonlinear and rectifying behavior. The room-temperature electroluminescence (EL) was observed under forward bias. The EL showed a weak white and strong yellowish emission colors (~575 nm) in the undoped ZnO/p-GaN:Mg heterojunctions before and after annealing process, respectively.
Recently, a large number of clinical experiments have shown that exposure of organic pollutants lead to various cancers through the abnormal cell growth. Environmental pollutants, such as 2, 3, 7, 8-Tetrachloro dibenzo-p-dioxin (TCDD) and polycyclic aromatic hydrocarbons (PAHs), are carcinogen and are known to cause the cognitive disability and motor dysfunction in the developing of brain. The effects of these pollutants on neurodevelopmental disorder is well established, but the underlying mechanism(s) and similarity of gene expression profiles in human brain tumors with organic pollutants still remain unclear. In this study, we first examined the gene expression profiles in glioblastomas compared with meningioma that are kinds of primary human brain tumor by using human cDNA microarray. The results of cDNA microarray analysis revealed that 26 genes were upregulated (Z-ratio>2.0) and 14 genes were downregulated (Z-ratio<-2.0) in glioblastoma compared with meningioma. From the altered gene patterns, mitogen-activated protein kinase (MAPK) signaling related genes, such as MAP2K3, MAP3K11 and jun activated domain binding protein, and transcription factors, such as UTF2 and TF12, were upregulated in glioblastoma. Also, we tried to investigate the relation between important genes up- and down-regulated in giloblastoma and various organic pollutants. Therefore, the identification of changes in the patterns of gene expression may provide a better understanding of the molecular mechanisms involved in human primary brain tumors and of the relation between gene expression profiles and organic pollutants in brain tissue.
Nonsteroidal anti-inflammatory drugs (NSAIDs), particularly the highly selective cyclooxygenase (COX)-2 inhibitors have been shown to decrease the growth of tumor, in part, by inhibition of neovascularization. Recently, besides mature endothelial cells, endothelial progenitor cells (EPCs) have been shown to contribute neovascularization in angiogenic tissues. In this study, we addressed a question whether nimesulide, a selective COX-2 inhibitor, could affect differentiation of EPCs into adhesive endothelial cells in vitro. Total mononuclear cells were isolated from cord blood by Ficoll density gradient centrifugation, and then the cells were incubated with nimesulide or vehicle control for 7 days. The number of adherent and spindle-shaped cells decreased by nimesulide treatment in a concentration-dependent fashion at a concentration range of 5 - 200 ${\mu}M$. Moreover, the adherent cells double positive for DiI-ac-LDL uptake and lectin binding significantly decreased upon nimesulide treatment. There was no change of expression of CD31 between treatment and control groups, whereas slight reduction was detected upon treatment in expression of VE-cadherin, ICAM-1, vWF, ${\alpha}v$, and ${\alpha}5$. Nimesulide also reduced cell viability during first 3 days' culture and induced apoptosis in adherent EPCs, resulting in increased annexin-V-positive and propidium iodide-negative cells. Taken together, these results suggest that nimesulide could be applied for the inhibition of new vessel formation, in part, by inhibiting differentiation and survival of EPCs.
Long-chain polyunsaturated fatty acids (LC-PUFA) promote the development of brain and vision of the fetus, relieve inflammation, inhibit oral dysplasia of rumor cell, decrease the incidence of cardiovascular disease and regulate arrhythmia. ${\Delta}-6$ desaturase is the rate-limited enzyme in the desaturation process. This study reports the cloning, characterization and tissue expression of a ${\Delta}-6$ desaturase gene in the chicken. PCR primers were designed based on the predicted sequence of chicken ${\Delta}-6$ desaturase (accession number: XM421053) and used to isolate a cDNA fragment of 1,323 bp from chicken liver. Based on the 1,323 bp fragment an EST (BI390105) was obtained by BLAST. The EST and 5'nd of the 1,323 bp fragment were partially overlapped. Gene specific primers derived from the EST were used for amplification of the 5'nd. Another gene-specific primer derived from the 1,323 bp fragment was used for amplification of the 3'nd by 3'ACE. Then the three overlapping cDNA sequences obtained were assembled with DNAMAN software and a full-length ${\Delta}-6$ desaturase of 2,153 bp was obtained. The full-length cDNA contained an ORF of 1,335 bp with a 5'ntranslated region of 147 nucleotides followed by an ATG initiation codon. Stop codon TGA was at position 1,481-1,483 bp. The deduced amino acids shared an homology above 77% with bovine, mice, orangutan, rat and human. The protein sequence had three histidine-rich regions HDFGH (HisI region), HFQHH (HisII region) and HH (HisIII region), a cytochrome $b_{5}$-like domain containing a heme-binding motif and two transmembrane domains. Sequence analysis of the chicken genomic DNA revealed that the coding sequence of chicken ${\Delta}-6$ desaturase included 12 exons and 11 introns. Semi-quantitative RT-PCR showed that the ${\Delta}-6$ desaturase expression levels were in turn liver, spleen, pancreas, lung, breast muscle, heart, and abdominal fat. The expression of ${\Delta}-6$ desaturase in liver was significantly higher than that in breast muscle (p<0.01). The expression of ${\Delta}-6$ desaturase in lung was significantly higher than that in abdominal fat (p<0.01). This is the first clone of chicken ${\Delta}-6$ desaturase.
This study was performed to develop the biological treatment technology of wastewater polluted with heavy metals. The copper-tolerant bacteria, Pseudomonas stutzeri which possessed the ability to accumulate copper, was isolated from mine wastewaters polluted with various heavy metals. The characteristics of copper accumulation in the cells and the recovery of the copper from the cells accumulating zinc, were investigated. Removal rate of copper from the solution containing 100mg/l of copper by copper-tolerant bacteria was more than 78% at 2 days after inoculation with the cells. A large number of the electron-dense granules were found mainly on the cell wall and cell membrane fractions, when determined by transmission electron microscopy. Energy dispersive X-ray spectroscopy revealed that the electron-dense granules were copper complex with the substances binding copper. The copper accumulated into the cells was not desorbed by deistilled water, but more than 80% of the copper accumulated was desorbed by 0.1M-EDTA solution. The residues of the cells after combustion at $550^{\circ}C$ amounted to about 23.2% of the dry weight of the cells. EDS analysis showed that residues were relatively pure copper compound containing more than 78.4% of copper.
Objectives : It has long been known about the anticancer effect of GRR-HAS, however, it has not been systemically determined the differentially regulated genes by GRR-HAS in cancer cells. The purpose of this study is to screen the GRR-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cell lines. Oligonucleotide microarray and proteomic approaches were employed to screen the differential expression genes. Methods : GRR~HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of GRR-HAS (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of GRR-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cells in all concentrations(0.1, 0.5, 1.5, 10,$20mg/m{\ell}$). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 320 with 6 up-regulated and 314 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. One down -regulated protein was protein disulfide isomerase and up-regulated proteins were fatty acid binding protein 1 and 14-3-3 gan1lTIa protein by $1.5mg/m{\ell}$ of CRR-HAS. Discussion : This study showed the comprehensive gene expression analysis using oligonucleotide microarray for the screening of GRR-HAS mediated differentially regulated genes. These results will provide a better application of GRR-HAS in cancer field and drug target development.
A number of studies on eco-friendly and healthy building materials are being conducted as modern people are becoming more conscious about health and the environment they live in. Among those materials, studies on Hwangtoh are the most prevalent but due to its strength, crack coming from drying shrinkage, and susceptibility to water, the usage of Hwangtoh is incomplete and limited to be used as a common building material. Cement concrete, considered as one of the most widely used building materials, is extensively used in construction because it is economical, easily accessible and moldable and has proper compressive strength. Due to carbon dioxide created in the process of making cement concrete, it is recognized as pollution. Accordingly, there are a lot of studies on reduction of carbon dioxide in cement concrete industry. There are increasing numbers of researches as well as developments on Hwangtoh or traditional construction materials used in South Korea to reduce the environmental problems. Therefore, this study suggests the basic features of the construction material that can replace cement concrete in the future with the non-sindtered cement mixed with non-sintering hwangtoh which is made with the furnace slag and multiple stimulants.
This study was designed to assess the iron nutritional status and anemia of high school students. 383 female subjects in Ulsan Metropolitan city were evaluated using a questionnaire, and a measurement of hematological indices. The average height and weight of the respondents were 161.24$\pm$4.90 cm and 53.12$\pm$6.37kg, repectively. The average BMI(body mass index) was 20.43$\pm$2.26 which was in the normal ramge. The average hemoglobin(Hb) concentration of the subjects was 13.14$\pm$0.97g/dl, and the average hematocrit(Hct) level was 40.84$\pm$17.40%. Transferrin saturation{TS(%)} was 20.86$\pm$10.32%, and the ferritin by Hct(<36%), 27.2%by TS(<14%),26.6% by ferritin(<12 ng/ml). As for clinical symptoms, the greatest number of respondents reported that they experienced ‘decreased ability to concectrate’. Mean daily intakes of iron were 14.89$\pm$4.48 mg and heme iron intakes were 5.04$\pm$2.13 mg, which was 29.6% of total iron intake. The total iron binding capacity(TIBC) was negatively correlated with Hb concentration(r= -0.222, p<0.01). Serum ferritin was positively correlated with Hb concentration(r= - 0.323, p<0.05) and negatively correlated with TIBC(r= -0.367, p<0.01). TS(%) was positively correlated with Hb concentration(r= 0.402, p<0.01) and positively correlated with serum ferritin(r=0.413, p<0.01). As for the correlation between blood biochemistry and clinical symptoms related to anemia, the Hb concentration was negatively correlated with ‘shortening of breath when going upstairs(p>0.05)’ and ‘cold hands and feet’ significantly(r= -0.109, p<0.05). The level of Mean corpuscular volume(MCV) was negatively correlated with ‘feel dizzy when standing up’,‘tired out easily’, and ‘decrease ability to concentrate’ significantly(p<0.05). In particular, the level of Fe was negatively correlated with ‘shortening of breath when going upstairs’ and ‘feeling blue’ significantly(p<0.01). These results suggest That the prevalence of iron deficiency of female high school students is very high, therefore guidelines for diet supports and nutrition education to improve their iron status should be provided.
Metallothionein (MT) is a low molecular weight, cysteine-rich, metal-binding, and non-enzymatic protein. The present study was carried out to investigate the expression of MT gene as well as the localization of MT in regenerating rat liver. In partial hepatectomized rats, MT mRNA was detected as early as 1 hr and reached a maximal level by 8 hr after the operation. Thereafter, this level decreased gradually until 24 hr, and it became similar to that of sham control. Meanwhile, time course of MT immunoreactivity using immunogold-labelling revealed that the number of gold particles in hepatocytes increased significantly by 12 hr, but decreased at 48 hr after partial hepatectomy. Ultrastructurally, immunogold particles indicating the presence of MT were distributed in both the cytoplasm and the nucleus of the rat hepatocytes, particularly the particles were distributed at rough endoplasmic reticulum and nucleolus and did not seem to adhere to mitochondria or lysosomes in proliferating hepatocytes. Briefly, high level of MT mRNA expression and the intense immunoreactivity and/or the specific localization of MT was observed during liver regeneration. These results suggest that MT possibly involves hepatocyte proliferation via the storage or the supply of various metal ions in the regenerating rat liver.
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