• Journal of Microbiology and Biotechnology
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• v.18 no.8
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• pp.1364-1367
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• 2008

Oxidative stress damages all cellular constituents, and therefore, cell has to possess various defense mechanisms to cope. Saccharomyces cerevisiae, widely used as a model organism for studying cellular responses to oxidative stress, contains three glutathione peroxidase (Gpx) proteins. Among them, Gpx3 plays a major defense role against oxidative stress in S. cerevisiae. In this study, in order to identity the new interaction proteins of Gpx3, we carried out two-dimensional gel electrophoresis after immunoprecipitation (IP-2DE), and MALDI-TOF mass spectrometry. The results showed that several proteins including protein disulfide isomerase, glutaredoxin 2, and SSY protein 3 specifically interact with Gpx3. These findings led us to suggest the possibility that Gpx3, known as a redox sensor and ROS scavenger, has another functional role by interacting with several proteins with various cellular functions.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.4
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• pp.1667-1674
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• 2012

Objective: To explore the differential protein expression profile in EC304 gastric cancer cells induced by alphastatin. Methods: Cultured EC304 cells in the exponential phase of growth were randomly divided into alphastatin and control groups. Total proteins were extracted and the two dimensional electrophoresis (2-DE) technique was applied to analyze differences in expression with ImageMaster 2D Platinum 5.0 software. Proteins were identified using the MASCOT database and selected differently expressed proteins were characterised by western blotting and immunofluorescence. Results: $1350{\pm}90$ protein spots were detected by the ImageMaster software in the 2-DE gel images from the control and alphastatin groups. The match rate was about 72-80% for the spectrum profiles, with 29 significantly different protein spots being identified, 10 upregulated, 16 downregulated, two new and one lost. The MASCOT search scores were 64-666 and the peptide matching numbers were 3-27 with sequence coverage of 8-62%. Twenty-three proteins were checked by mass spectrometry, including decrease in Nm23 and profilin-2 isoform b associated with the regulation of actin multimerisation induced by extracellular signals. Conclusion: The proteome in EC304 cells is dramatically altered by alphastatin, which appears to play an important role in modulating cellular activity and anti-angiogenesis by regulating protein expression and signal transduction pathways through Nm23 and profilin-2 isoform b, providing new research directions for anti-angiogenic therapy of gastric cancer.

• Journal of the Institute of Convergence Signal Processing
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• v.9 no.1
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• pp.1-9
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• 2008

One of the problems for implementing the spot detection phase in the 2-DE gel image analysis program is the eliminating noises in the image. Remained noises after the preprocessing phase cause the over-segmented regions by the segmentation phase. To identify and exclude the over-segmented background regions, if we use the fixed thresholding method that is choosing an intensity value for the threshold, the spots that is invisible by the eyes but mean a very small amount proteins which have important role in the biological samples could be eliminated. This paper propose an adaptive thresholding method that come from an idea that is got on statistical analysing for the prominences of the peaks. The adaptive thresholding method works as following. Firstly we calculate an average prominence value curve and fit it to exponential function curve, as a result we get parameters for the exponential function. And then we calculate a threshold value by using the parameters and probability distribution of errors. Lastly we apply the threshold value to the region for determining the region is a noise or not. According to the probability distribution of errors, the reliability is 99.85% and we show the correctness of the proposed method by representing experiment results.

• Journal of Microbiology
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• v.44 no.4
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• pp.369-374
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• 2006

To investigate the mechanism of salt tolerance of gram-positive moderately halophilic bacteria, two-dimensional gel electrophoresis (2-D PAGE) was employed to achieve high resolution maps of proteins of Halobacillus dabanensis $D-8^T$. Approximately 700 spots of proteins were identified from these 2-D PAGE maps. The majority of these proteins had molecular weights between 17.5 and 66 kDa, and most of them were distributed between the isoelectric points (pI) 4.0 and 5.9. Some protein spots were distributed in the more acidic region of the 2-D gel (pI <4.0). This pattern indicated that a number of proteins in the strain $D-8^T$ are acidic. To understand the adaptation mechanisms of moderately halophilic bacteria in response to sudden environmental changes, differential protein profiles of this strain were investigated by 2-D PAGE and $Imagemaster^{TM}$ 2D Platinum software after the cells were subjected to salt shock of 1 to 25% salinity for 5 and 50 min. Analysis showed 59 proteins with an altered level of expression as the result of the exposure to salt shock. Eighteen proteins had increased expression, S proteins were induced, and the expression of 33 proteins was down-regulated. Eight of the up-regulated proteins were identified using MALDI-TOF/MS and MASCOT, and were similar to proteins involved in signal transduction, proteins participating in energy metabolism pathways and proteins involved in stress.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.470-478
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• 2012

Recent genome comparisons of E. coli B and K-12 strains have indicated that the makeup of the cell envelopes in these two strains is quite different. Therefore, we analyzed and compared the envelope proteomes of E. coli BL21(DE3) and MG1655. A total of 165 protein spots, including 62 nonredundant proteins, were unambiguously identified by two-dimensional gel electrophoresis and mass spectrometry. Of these, 43 proteins were conserved between the two strains, whereas 4 and 16 strain-specific proteins were identified only in E. coli BL21(DE3) and MG1655, respectively. Additionally, 24 proteins showed more than 2-fold differences in intensities between the B and K-12 strains. The reference envelope proteome maps showed that E. coli envelope mainly contained channel proteins and lipoproteins. Interesting proteomic observations between the two strains were as follows: (i) B produced more OmpF porin with a larger pore size than K-12, indicating an increase in the membrane permeability; (ii) B produced higher amounts of lipoproteins, which facilitates the assembly of outer membrane ${\beta}$-barrel proteins; and (iii) motility- (FliC) and chemotaxis-related proteins (CheA and CheW) were detected only in K-12, which showed that E. coli B is restricted with regard to migration under unfavorable conditions. These differences may influence the permeability and integrity of the cell envelope, showing that E. coli B may be more susceptible than K-12 to certain stress conditions. Thus, these findings suggest that E. coli K-12 and its derivatives will be more favorable strains in certain biotechnological applications, such as cell surface display or membrane engineering studies.

• Microbiology and Biotechnology Letters
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• v.39 no.1
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• pp.9-19
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• 2011

The Bacillus subtilis pyrimidine biosynthetic (pyr) operon encodes all of the enzymes for the de novo biosynthesis of Uridine monophosphate (UMP) and additional cistrones encoding a uracil permease and the regulatory protein PyrR. The PyrR is a bifunctional protein with pyr mRNA-binding regulatory funtion and uracil phosphoribosyltransferase activity. To study the global regulation by the pyrR deletion, the proteome comparison between Bacillus subtilis DB104 and Bacillus subtilis DB104 ${\Delta}$pyrR in the minimal medium without pyrimidines was employed. Proteome analysis of the cytosolic proteins from both strains by 2D-gel electrophoresis showed the variations in levels of protein expression. On the silver stained 2D-gel with an isoelectric point (pI) between 4 and 10, about 1,300 spots were detected and 172 spots showed quantitative variations in which 42 high quantitatively variant proteins were identified. The results showed that production of the pyrimidine biosynthetic enzymes (PyrAA, PyrAB, PyrB, PyrC, PyrD, and PyrF) were significantly increased in B. subtilis DB104 ${\Delta}$pyrR. Besides, proteins associated carbohydrate metabolism, elongation protein synthesis, metabolism of cofactors and vitamins, motility, tRNA synthetase, catalase, ATP-binding protein, and cell division protein FtsZ were overproduced in the PyrR-deficient mutant. Based on analytic results, the PyrR might be involved a number of other metabolisms or various phenomena in the bacterial cell besides the pyrimidine biosynthesis.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.40 no.2
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• pp.236-244
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• 1995

This study was carried out to clarify the changes in dry matter weight and physicochemical property of job's tears grain during ripening period. Four varieties of job's tears were seeded at April 20, May 10, and May 30, andgrains were harvested at every 7 days from heading to maturity, The whole seed weight of thick hull varieties, Kimje and Aewon, was higher than that of improved thin hull varieties, Suwon 3 and Suwon 6, from heading to maturity, However, the dehulled grain weight of thin hull variety were higher than that of thick hullvariety from 35 days after heading. The first heading spikelet showed lower 100-grain weight com-pared with the spikelets headed at full blooming stage among the same plant. The increased rate of de hulled grain weight during maturing period was higher in thin hull variety than thick hull one, later seeding date than early seeding, and intermediate headed spikelet than early head-ed one, respectively. As maturing proceeds the protein content of de hulled grain was decreased, alkali digestibility value(ADV) showed a rising trend, and little change was found in gel consistency of milled grain flour. And same trends were found in protein content, ADV and gel consistency as the seeding date was delayed. Among amylogram properties of job's tears flour the maximum, minimum, and final viscosity as well as breakdown were decreased, and set back was increased in later seeding plots

• KSBB Journal
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• v.14 no.2
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• pp.146-152
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• 1999

Screening experiments were carried out in order to select bacteria causing brown blotch disease on the mushroom, Pleurotus ostreatus. Four bacteria causing brown blotch disease were isolated from Pleurotus ostreatus and soils around the mushroom farm. Three strains showing antagonism against brown blotch causing bacteria, A-11, A-20 and A-29 were also isolated through methods pitting test, cross checking and biochemical test, and identified as Pseudomonas fluorescence for A-11 and A-20, and Pseudomonas sp. for A-29, respectively. Colonial morphology test also showed that A-11 and A-29 were appeared as transparent gel with green color, whereas the colony of A-20 showed opaque gel with light green color.

• Korean Journal of Materials Research
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• v.26 no.12
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• pp.757-763
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• 2016

$Ho^{3+}/Yb^{3+}/Tm^{3+}$ tri-doped $NaY_{1-x}(WO_4)_2$ phosphors with proper doping concentrations of $Ho^{3+}$, $Yb^{3+}$ and $Tm^{3+}$ ($x=Ho^{3+}+Yb^{3+}+Tm^{3+}$, $Ho^{3+}$=0.04, 0.03, 0.02, 0.01, $Yb^{3+}$=0.35, 0.40, 0.45, 0.50 and $Tm^{3+}$=0.01, 0.02, 0.03, 0.04) were successfully synthesized via the microwave sol-gel route, and their upconversion properties were investigated. Well-crystallized microcrystalline particles showed fine and homogeneous microcrystalline morphology with particle sizes of $1-2{\mu}m$. The optical properties were comparatively examined using photoluminescence emission and Raman spectroscopy. Under excitation at 980 nm, the doped particles exhibited white emissions based on blue, green and red emission bands, which correspond to the $^1G_4{\rightarrow}^3H_6$ transitions of $Tm^{3+}$ in the blue region, the $^5S_2/^5F_4{\rightarrow}^5I_8$ transitions of $Ho^{3+}$ in the green region, the $^5F_5{\rightarrow}^5I_8$ transitions of $Ho^{3+}$, and the $^1G_4{\rightarrow}^3F_4$ and $^3H_4{\rightarrow}^3H_6$ transitions of $Tm^{3+}$ in the red region. The pump power dependence of the upconversion emission intensity and the Commission Internationale de L'Eclairage chromaticity coordinates of the phosphors were evaluated in detail.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.16
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• pp.7123-7127
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• 2015

Purpose: To determine the utility of secondary circulating prostate cells for predicting early biochemical failure after radical prostatectomy for prostate cancer and compare the results with the Walz nomagram. Materials and Methods: A single centre, prospective study of men with prostate cancer treated with radical prostatectomy between 2004 and 2014 was conducted, with registration of clinical-pathological details, total serum PSA pre-surgery, Gleason score, extracapsular extension, positive surgical margins, infiltration of lymph nodes, seminal vesicles and pathological stage. Secondary circulating prostate cells were obtained using differential gel centrifugation and assessed using standard immunocytochemistry with anti-PSA. Biochemical failure was defined as a PSA >0.2ng/ml, predictive values werecalculated using the Walz nomagram and CPC detection. Results: A total of 326 men participated, with a median follow up of 5 years; 64 had biochemical failure within two years. Extracapsular extension, positive surgical margins, pathological stage, Gleason score ${\geq}8$, infiltration of seminal vesicles and lymph nodes were all associated with higher risk of biochemical failure. The discriminative value for the nomogram and circulating prostate cells was high (AUC >0.80), predictive values were higher for circulating prostate cell detection, with a negative predictive value of 99%, sensitivity of 96% and specificity of 75%. Conclusions: The nomagram had good predictive power to identify men with a high risk of biochemical failure within two years. The presence of circulating prostate cells had the same predictive power, with a higher sensitivity and negative predictive value. The presence of secondary circulating prostate cells identifies a group of men with a high risk of early biochemical failure. Those negative for secondary CPCs have a very low risk of early biochemical failure.