• Journal of Korean Neurosurgical Society
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• v.59 no.6
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• pp.544-550
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• 2016

Objective : Cerebral endothelial cells have unique biological features and are fascinating candidate cells for stroke therapy. Methods : In order to understand the molecular mechanisms of human cerebral endothelial cell (hCMEC/D3) transplantation in a rat stroke model, we performed proteomic analysis using 2-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Protein expression was confirmed by quantitative real-time PCR and Western blot. Results : Several protein spots were identified by gel electrophoresis in the sham, cerebral ischemia (CI), and CI with hCMEC/D3 treatment cerebral ischemia with cell transplantation (CT) groups, and we identified 14 differentially expressed proteins in the CT group. Proteins involved in mitochondrial dysfunction (paraplegin matrix AAA peptidase subunit, SPG7), neuroinflammation (peroxiredoxin 6, PRDX6), and neuronal death (zinc finger protein 90, ZFP90) were markedly reduced in the CT group compared with the CI group. The expression of chloride intracellular channel 4 proteins involved in post-ischemic vasculogenesis was significantly decreased in the CI group but comparable to sham in the CT group. Conclusion : These results contribute to our understanding of the early phase processes that follow cerebral endothelial cell treatment in CI. Moreover, some of the identified proteins may present promising new targets for stroke therapy.

• Animal Systematics, Evolution and Diversity
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• v.11 no.2
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• pp.207-221
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• 1995

Electrophoretic analysis was performed to determine the interspecific genetic relationships of the genus Raja collected in Korea waters. We confirmed 5 species [Raja (Okamejei) meerdervoortii Bleeker, R.(O).acutispina Ishiyama, R.(O) kenojei Muller et Henle, Raja (Dipturus) kwangtungensis Chu, and R.(D.) pulchra Liu] inhabting in Korea. As results of starch gel electrophoresis, 24 genetic loci were obtained from 13 enzymic and non-enzymic proteins, and allele frequencies of each locus were calculated . Even thought their various dorsal color patterns, the mean genetic similarity among 4 conspecific populations of R.(O) kenojei showed highly close genetic relationships (S=0.966) R.(O) kenojei and R.(D) kwangtungensis which belong to different subgenus , revealed subspecific level of differentiation (S=0.829). R. (Diptuuus ) kwangtungensis was genetically much closer to Raja(Okamejei) species than to Raja (Dipturus) species group. R.(O) acutispina and R.(D) kwangtungensis were unrecorded species to Korea.

• YAKHAK HOEJI
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• v.54 no.1
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• pp.32-38
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• 2010

The aim of this study was to evaluate the effect of galangin towards hydrogen peroxide-induced DNA damage. The calf thymus DNA and Chinese Hamster Lung (CHL) cells were used to measure 8-hydroxy-2'-deoxyguanosine(8-OH2'dG) as an indicator of DNA oxidative damage using high performance liquid chromatography with electrochemical detection. Hydrogen peroxide in the presence of Fe(II) ion induced the formation of 8-OH2'dG in both calf thymus DNA and CHL cells. The DNA damage effects were enhanced by increasing the concentration of Fe(II) ion and inhibited by galangin. In the single cell gel electrophoresis (Comet assay), galangin and dl-a-tocopherol showed an inhibitory effect in CHL on hydrogen peroxide induced DNA single strand breaks. Galangin showed more potent activity than dl-$\alpha$-tocopherol under our experimental conditions. These results indicate that galangin can modify the action mechanisms of the oxidative DNA damage and may act as chemopreventive agents against oxidative stress.

• Journal of Life Science
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• v.18 no.6
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• pp.814-825
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• 2008

Ascochlorin (ASC) is prenyl-phenol compound that was isolated from the fungus Ascochyta viciae. ASC reduces serum cholesterol and triglyceride levels, and suppresses hypertension, tumor development, ameliorates type I and II diabetes. Here, to better understand the mechanisms by which ASC regulates physiological or pathological events and induces responses in the pharmacological treatment of inflammation, we performed differential analysis of the proteome of the mouse macrophage RAW264.7 cells in response to ASC. In this study, we used a proteomic analysis of LPS-induced RAW264.7 cells treated by ASC, to identify proteins potentially involved in inflammatory processes. The RAW264.7 cell proteomes with and without treatment with ASC were compared using two-dimensional electrophoresis (2-D SDS-PAGE), matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS) and bioinformatics. The largest differences in expression were observed for the calreticulin (4-fold decrease), ${\beta}-actin$ (4-fold decrease) and vimentin (1.5-fold decrease). In addition, rabaptin was increased 3-fold in RAW264.7 cells treated with ASC. The expression of some selected proteins was confirmed by RT-PCR analysis.

• Journal of Applied Biological Chemistry
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• v.50 no.2
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• pp.58-62
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• 2007

Identification of Dual-specificity protein phosphatase (DSP) substrates is essential in revealing physiological roles of DSPs. We isolated VHZ-interacting proteins from extracts of 293T cells overexpressing a VHZ (C95S, D65A) mutant known to be substrate- trapping mutant. Analysis of specific proteins bound to VHZ by 2D gel electrophoresis and mass spectroscopy revealed that these proteins contained Chaperonin containing TCP1, Type II phosphatidylinositol phosphate kinase ${\gamma}$, Intraflagellar transport 80 homolog, and Kinesin superfamily protein 1B. VHZ-interacting proteins showed that VHZ is involved in many important cellular signal pathways such as protein folding, molecular transportation, and tumor suppression.

• The KIPS Transactions:PartD
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• v.12D no.5 s.101
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• pp.719-728
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• 2005

2-dimensional electrophoresis(2DE) is a separation technique to identify proteins contained in a sample. However, the image is very sensitive to its experimental conditions as well as the quality of scanning. In order to adjust the possible variation of spots in a particular image, a user should manually annotate landmark spots on each gel image to analyze the spots of different images together. However, this operation is an error-prone and tedious job. This thesis develops an automated method of extracting the landmark spots of an image based on landmark profile. The landmark profile is created by clustering the previously identified landmarks of sample images of the same type. The profile contains the various properties of clusters identified for each landmark. When the landmarks of a new image need to be fount all the candidate spots of each landmark are first identified by examining the properties of its clusters. Subsequently, all the landmark spots of the new image are collectively found by the well-known optimization algorithm $A^*$. The performance of this method is illustrated by various experiments on real 2DE images of mouse's brain-tissues.

• BMB Reports
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• v.38 no.5
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• pp.545-549
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• 2005

Acidithiobacillus ferrooxidans is one of the most important bacterium used in bioleaching, and can utilize $Fe^{2+}$ or sulphide as energy source. Growth curves for Acidithiobacillus ferrooxidans under phosphate starvation and normal condition have been tested, showing lag, logarithmic, stationary and aging phases as seen in other bacteria. The logarithmic phases were from 10 to 32 hours for Acidithiobacillus ferrooxidans cultivated with normal cultivating condition and from 20 to 60 hrs for Acidithiobacillus ferrooxidans cultivated phosphate starvation. Differences of protein patterns of Acidithiobacillus ferrooxidans growing in case of normal or phosphate starvation were separately investigated after cultivation at $30^{\circ}C$ by the analysis of two-dimensional gel electrophoresis (2-DE), matrix-assisted laser desorption/ionization (MALDI)-Mass spectrometry. There were total 6 protein spots identified, which were Recombination protein recA, RNA helicase, AP2 domain-containing transcription factor, NADH dehydrogenase I chain D, Hyothetical protein PF1669, and Transaldolase STY3758. From the 6 identified protein spots, 3 proteins were found to be decreased in expression at the cultivating condition of phosphate starvation, while another three upregulated.

• Reproductive and Developmental Biology
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• v.32 no.4
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• pp.255-261
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• 2008

The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield-the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy-specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non-pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non-pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH $3.0{\sim}10.0$ strip, by loading a 2-mg milk protein sample. After the second-dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non-pregnant and pregnant cattle milk protein spots, using ImageMaster, this was followed by analysis with MALDI TOF-MS. Analysis of the 2-DE gel image resulted in a total of approximately $500{\sim}600$ protein spots, of which 12 spots were differentially expressed, six spots were up-regulated, and four spots were down-regulated; two spots were identified as pregnancy-specific proteins. These proteins were identified as lactoferrin, NA-DH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2-D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

• Korean Journal of Microbiology
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• v.31 no.2
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• pp.166-174
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• 1993

Streptomyces coeUc%r (Muller) cells were treated with $100 \mu$M hydrogen peroxide for I hour and proteins synthesized during hydrogen peroxide stress were labeled with L-[$^{35}S$]-methionine. Total cellular proteins were extracted and analyzed by two-dimensional polyacrylamide gel electrophoresis. In exponential growth phase, synthesis of about 100 proteins was increased by hydrogen peroxide treatment. These proteins were named as Pin (£eroxide-inducib]e) proteins and classified into 4 subgroups according to their induction time after hydrogen peroxide treatment. About 60 of them were found to be induced within 20 minutes and maintained throughout I hour of treatment. In stationary growth phase. synthesis of 62 proteins was increased by hydrogen peroxide and 21 of them were the same Pins found in exponential growth phase. Proteins from the mutants which are resistant to hydrogen peroxide were obtained in exponential growth phase and compared with those from the wild type on two-dimensional gel. The three mutants, N7, N9. and N24, were found to have higher constitutive leve]s of ]5, 17, and 15 Pin proteins respectively, than the wild type. 9 of these Pin proteins (D74.7a, E76.0c, E23.3. F50.7, F47.2a. F25.5, G39.6b, G24.0, H39.6a) increased in two of the three mutants and 3 proteins (F39.7, H6I.7. 120.8) increased in all of the three mutants. These proteins might play important roles in the response of S. coelic%r to hydrogen peroxide.

• Rapid Communication in Photoscience
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• v.3 no.4
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• pp.76-78
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• 2014

Fluorescent nucleic acids were prepared utilizing the polymerase extension (PEX) reaction to incorporate fluorescent molecules. 2'-Deoxyuridine triphosphate (dUTP) derivatives possessing pyrene molecules as fluorophores were synthesized using the aqueous-phase Sonogashira coupling between 5-Iodo-dUTP and acetylene-linked pyrene molecules. The incorporation of the pyrene (Py)-labeled deoxyuridine triphosphates (PyU) into DNA by polymerase was evaluated by polyacrylamide gel electrophoresis, demonstrating that the PyU can work as a good substrate for the PEX reaction. The fluorescent properties of the functionalized DNA prepared by the PEX reaction were characterized by steady-state fluorescence measurements. The Py-conjugated DNA showed typical emission spectra of the pyrene, and the DNA with two pyrene molecules connected to each other by a diethylene glycol linker exhibited a broadened emission attributed to the electronic interaction between the Py molecules.