• Title/Summary/Keyword: Mass Matrix

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Nano-scale Proteomics Approach Using Two-dimensional Fibrin Zymography Combined with Fluorescent SYPRO Ruby Dye

  • Choi, Nack-Shick;Yoo, Ki-Hyun;Yoon, Kab-Seog;Maeng, Pil-Jae;Kim, Seung-Ho
    • BMB Reports
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    • v.37 no.3
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    • pp.298-303
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    • 2004
  • In general, a SYPRO Ruby dye is well known as a sensitive fluorescence-based method for detecting proteins by one-or two-dimensional SDS-PAGE (1-DE or 2-DE). Based on the SYPRO Ruby dye system, the combined two-dimensional fibrin zymography (2-D FZ) with SYPRO Ruby staining was newly developed to identify the Bacillus sp. proteases. Namely, complex protein mixtures from Bacillus sp. DJ-4, which were screened from Doen-Jang (Korean traditional fermented food), showed activity on the zymogram gel. The gel spots on the SYPRO Ruby gel, which corresponded to the active spots showing on the 2-D FZ gel, were analyzed by a matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometric analysis. Five intracellular fibrinolytic enzymes of Bacillus sp. DJ-4 were detected through 2-D FZ. The gel spots on the SYPRO Ruby dye stained 2-D gel corresponding to 2-D FZ were then analyzed by MALID TOF MS. Three of the five gel spots proved to be quite similar to the ATP-dependent protease, extracellular neutral metalloprotease, and protease of Bacillus subtilis. Also, the extracellular proteases of Bacillus sp. DJ-4 employing this combined system were identified on three gels (e.g., casein, fibrin, and gelatin) and the proteolytic maps were established. This combined system of 2-D zymography and SYPRO Ruby dye should be useful for searching the specific protease from complex protein mixtures of many other sources (e.g., yeast and cancer cell lines).

Geometrical Reorientation of Distorted Road Sign using Projection Transformation for Road Sign Recognition (도로표지판 인식을 위한 사영 변환을 이용한 왜곡된 표지판의 기하교정)

  • Lim, Hee-Chul;Deb, Kaushik;Jo, Kang-Hyun
    • Journal of Institute of Control, Robotics and Systems
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    • v.15 no.11
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    • pp.1088-1095
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    • 2009
  • In this paper, we describe the reorientation method of distorted road sign by using projection transformation for improving recognition rate of road sign. RSR (Road Sign Recognition) is one of the most important topics for implementing driver assistance in intelligent transportation systems using pattern recognition and vision technology. The RS (Road Sign) includes direction of road or place name, and intersection for obtaining the road information. We acquire input images from mounted camera on vehicle. However, the road signs are often appeared with rotation, skew, and distortion by perspective camera. In order to obtain the correct road sign overcoming these problems, projection transformation is used to transform from 4 points of image coordinate to 4 points of world coordinate. The 4 vertices points are obtained using the trajectory as the distance from the mass center to the boundary of the object. Then, the candidate areas of road sign are transformed from distorted image by using homography transformation matrix. Internal information of reoriented road signs is segmented with arrow and the corresponding indicated place name. Arrow area is the largest labeled one. Also, the number of group of place names equals to that of arrow heads. Characters of the road sign are segmented by using vertical and horizontal histograms, and each character is recognized by using SAD (Sum of Absolute Difference). From the experiments, the proposed method has shown the higher recognition results than the image without reorientation.

The Application of Genetic Algorithm for the Identification of Discontinuity Sets (불연속면 군 분류를 위한 유전자알고리즘의 응용)

  • Sunwoo Choon;Jung Yong-Bok
    • Tunnel and Underground Space
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    • v.15 no.1 s.54
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    • pp.47-54
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    • 2005
  • One of the standard procedures of discontinuity survey is the joint set identification from the population of field orientation data. Discontinuity set identification is fundamental to rock engineering tasks such as rock mass classification, discrete element analysis, key block analysis. and discrete fracture network modeling. Conventionally, manual method using contour plot had been widely used for this task, but this method has some short-comings such as yielding subjective identification results, manual operations, and so on. In this study, the method of discontinuity set identification using genetic algorithm was introduced, but slightly modified to handle the orientation data. Finally, based on the genetic algorithm, we developed a FORTRAN program, Genetic Algorithm based Clustering(GAC) and applied it to two different discontinuity data sets. Genetic Algorithm based Clustering(GAC) was proved to be a fast and efficient method for the discontinuity set identification task. In addition, fitness function based on variance showed more efficient performance in finding the optimal number of clusters when compared with Davis - Bouldin index.

Identification of Diapause and Non-diapause Associated Proteins in the Eggs of Multivoltine Silkworm Bombyx mori by MALDI MS Analysis

  • Saravanakumar, Ragunathan;Ponnuvel, Kangayam M.;Velu, Dhanikachalam;Koundinya, Panguluri R.;Sinha, Randir K.;Qadri, Syed M. Hussaini
    • International Journal of Industrial Entomology and Biomaterials
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    • v.16 no.2
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    • pp.37-48
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    • 2008
  • The diapause and non diapause associated proteins of multivoltine silkworm eggs were analysed by two dimensional (2D) gel electrophoresis. The study was made at 0 hr, 24 hrs and 48 hrs after oviposition. A total of four protein spots in diapause eggs at 24 hrs of oviposition and two protein spots in non diapause eggs at 0 hrs of oviposition were observed. All the six protein spots were considered to have association with diapause and non diapause characters. The molecular weight (MW) and isoelectric point (PI) of these 6 protein spots were calculated. The protein spots 1 and 2 observed in 0 hr of non diapause eggs were found to have the MW of 67 and 75 KDa and PI of 8.6 and 8.4 respectively. Similarly the four protein spots observed in diapause egg at 24 hrs of oviposition exhibited MW viz., 15, 17,20 and 25 KDa and PI of 5.3, 5.8, 6.5 and 6.0 respectively. All these 6 identified protein spots were subjected to in-gel digestion and resulted tryptic peptides were analyzed by Matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI TOF-MS). Databases searched based on experimentally determined molecular weights of peptides for the determination of the identities of proteins. The identified proteins indicated homology of 34% to 95%. The results indicate that the proteins may playa role in development of diapause and non diapause eggs.

Removal of Cs and Sr Ions by Absorbent Immobilized Zeolite with PVA (제올라이트를 PVA로 고정화한 흡착제에 의한 Cs과 Sr 이온 제거)

  • Lee, Chang-Han;Lee, Min-Gyu
    • Journal of Korean Society of Environmental Engineers
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    • v.37 no.8
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    • pp.450-457
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    • 2015
  • In this research a adsorbent, PVA-Zeolite bead, was prepared by immobilizing zeolite with PVA. The results of XRD and SEM analysis showed that the prepared PVA-Zeolite beads had porous structure and the zeolite particles were in mobilized within the internal matrix of the beads. The adsorption properties of Sr ion and Cs ion with the adsorbent were studied by different parameters such as effect of pH, adsorption rate, and adsorption isotherm. The adsorption of Sr ion and Cs ion reached equilibrium after 540 minutes. The adsorption kinetics of both ions by the PVA-Zeolite beads were fitted well by the pseudo-second-order model more than pseudo-first-order model. The equilibrium data fitted well with Langmuir isotherm model. The maximum adsorption capacities of Sr ion and Cs ion calculated from Langmuir isotherm model were 52.08 mg/g and 58.14 mg/g, respectively. The external mass transfer step was very fast compared to the intra-particle diffusion step in the adsorption process of Cs ion and Sr ion by the PVA-Zeolite beads. This result implied that the rate controlling step was the intra-particle diffusion step.

Cloned Placenta of Korean Native Calves Died Suddenly at Two Months after Birth Displays Differential Protein Expression

  • Kim Hong Rye;Kang Jae Ku;Lee Hye Ran;Yoon Jong Taek;Seong Hwan Hoo;Jung Jin Kwan;Park Chang Sik;Jin Dong Il
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.63-68
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    • 2005
  • Cloned calves derived from somatic cell nuclear transfer (SCNT) have been frequently lost by sudden death at 1 to 3 month following healthy birth. To address whether placental anomalies are responsible for the sudden death of cloned calves, we compared protein patterns of 2 placentae derived from SCNT of Korean Native calves died suddenly at two months after birth and those of 2 normal placentae obtained from AI fetuses. Placental proteins were separated using 2-Dimensional gel electrophoresis. Approximately 800 spots were detected in placental 2-D gel stained with coomassie-blue. Then, image analysis of Malanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between normal and SCNT placentae. In the comparison of normal and SCNT samples, 8 spots were identified to be up-regulated proteins and 24 spots to be down-regulated proteins in SCNT placentae, among which proteins were high mobility group protein HMG1, apolipoprotein A-1 precursor, bactenecin 1, tropomyosin beta chain, $H^+-transporting$ ATPase, carbonic anhydrase II, peroxiredoxin 2, tyrosine-rich acidic matrix protein, serum albumin precursor and cathepsin D. These results suggested that the sudden death of cloned calves might be related to abnormal protein expression in placenta.

Identification of Proteins Affected by Iron in Saccharomyces cerevisiae Using Proteome Analysis

  • Lieu Hae-Youn;Song Hyung-Seok;Yang Seung-Nam;Kim Jae-Hwan;Kim Hyun-Joong;Park Young-Doo;Park Cheon-Seok;Kim Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.946-951
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    • 2006
  • To study the effect of iron on Saccharomyces cerevisiae, whole-cell proteins of Saccharomyces cerevisiae were extracted and subjected to two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and differentially expressed proteins were identified. The proteins separated were further identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and were compared with a protein database. Of more than 300 spots separated by molecular weight and isoelectric points, 27 differentially expressed spots were identified. Ten proteins were found to be differentially expressed at high iron concentration. Triosephosphate isomerase (TPI), YDR533C hypothetical protein, superoxide dismutase (SOD), 60 kDa heat-shock protein (HSP60), pyruvate dehydrogenase beta subunit 1 (PDB1), and old yellow enzyme 2 (OYE2) were upregulated, whereas thiol-specific antioxidant (TSA), regulatory particle non-ATPase subunit 8 (RPN8), thiol-specific peroxiredoxin 1 (AHP1), and fructose-1, 6-bisphosphate adolase (FBA) were downregulated by iron. Based on the result, we propose that SOD upregulated by iron would protect the yeast from oxidative stress by iron, and that TSA downregulated by iron would render cells hypersensitive to oxidative stress.

Fungal Growth and Manganese Peroxidase Production in a Deep Tray Solid-State Bioreactor, and In Vitro Decolorization of Poly R-478 by MnP

  • Zhao, Xinshan;Huang, Xianjun;Yao, Juntao;Zhou, Yue;Jia, Rong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.6
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    • pp.803-813
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    • 2015
  • The growth of Irpex lacteus F17 and manganese peroxidase (MnP) production in a selfdesigned tray bioreactor, operating in solid-state conditions at a laboratory scale, were studied. The bioreactor was divided into three layers by three perforated trays. Agroindustrial residues were used both as the carrier of bound mycelia and as a nutrient medium for the growth of I. lacteus F17. The maximum biomass production in the bioreactor was detected at 60 h of fermentation, which was consistent with the CO2 releasing rate by the fungus. During the stationary phase of fungal growth, the maximum MnP activity was observed, reaching 950 U/l at 84 h. Scanning electron microscopy images clearly showed the growth situation of mycelia on the support matrix. Furthermore, the MnP produced by I. lacteus F17 in the bioreactor was isolated and purified, and the internal peptide sequences were also identified with mass spectrometry. The optimal activity of the enzyme was detected at pH 7 and 25℃, with a long half-life time of 9 days. In addition, the MnP exhibited significant stability within a broad pH range of 4-7 and at temperature up to 55℃. Besides this, the MnP showed the ability to decolorize the polymeric model dye Poly R-478 in vitro.

The Antibiosis Action and Rice-Induced Resistance, Mediated by a Lipopeptide from Bacillus amyloliquefaciens B014, in Controlling Rice Disease Caused by Xanthomonas oryzae pv. oryzae

  • Li, Shu Bin;Xu, Shi Ru;Zhang, Rui Ning;Liu, Yuan;Zhou, Ren Chao
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.748-756
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    • 2016
  • In the present study, a lipopeptide (named AXLP14) antagonistic to Xanthomonas oryzae pv. oryzae (Xoo) was obtained from the culture supernatant of Bacillus amyloliquefaciens B014. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis demonstrated that AXLP14 consisted of surfactin homologs. The minimum inhibition concentration and minimum bactericidal concentration of AXLP14 against Xoo were determined to be 1.25 and 2.50 mg/ml, respectively. At a concentration of 0.613 mg/ml, AXLP14 strongly inhibited the formation of Xoo biofilm. AXLP14 also inhibited the motility of Xoo in a concentration-dependent manner. Applying AXLP14 to rice seedlings significantly reduced the incidence and severity of disease caused by Xoo. In Xoo-infected rice seedlings, AXLP14 strongly and continuously up-regulated the expression of both OsNPR1 and OsWRKY45. In addition, AXLP14 effectively inhibited the Xoo-induced up-regulation of the expression of the abscisic acid biosynthesis gene OsNECD3 and the abscisic acid signalingresponsive gene OsLip9, indicating that AXLP14 may protect rice against Xoo-induced disease by enhancing salicylic acid defense and interfering with the abscisic acid response to virulence.

Identifying Ambient PM2.5 Sources and Estimating their Contributions by Using PMF : Separation of Gasoline and Diesel Automobile Sources by Analyzing ECs and OCs (PMF 모델을 이용한 미세분진의 오염원 확인과 기여도 추정 : 탄소성분을 이용한 휘발유 및 경유차량 오염원의 분리)

  • Lee, Hyung-Woo;Lee, Tae-Jung;Kim, Dong-Sool
    • Journal of Korean Society for Atmospheric Environment
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    • v.25 no.1
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    • pp.75-89
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    • 2009
  • The purpose of this study was to identify $PM_{2.5}$ sources and to estimate their contributions to the border of Yongin-Suwon area, based on the analysis of the $PM_{2.5}$ mass concentration and the associated inorganic elements, ions and carbon components. The contribution of $PM_{2.5}$ sources were estimated by using a positive matrix factorization (PMF) model to identify air emission sources. For this study, $PM_{2.5}$ samples were collected from May, 2007 to April, 2008. The inorganic elements were analyzed by an ICP-AES. The ionic components in $PM_{2.5}$ were analyzed by an Ie. The carbon components were also analyzed by DRI/OGC analyzer. After performing PMF modeling, a total of 12 sources were identified and their contributions were quantitatively estimated. The contributions from each emission source were as follows: 11.3% from oil combustion source, 3.4% from bus/highway source, 5.8% from diesel vehicle source, 4.7% from gasoline vehicle source, 8.8% from biomass burning source, 15.1 % from secondary sulfate, 5.2% from secondary nitrate source, 13.4% from industrial related source, 4.1% from Cl-rich source, 19.6% from soil related source, 1.0% from aged sea salt, and 7.4% from coal combustion source, respectively. This study provides basic information on the major sources affecting air quality, and then it will help to effectively control $PM_{2.5}$ in this study area.